bims-auttor Biomed News
on Autophagy and mTOR
Issue of 2022‒04‒24
75 papers selected by
Viktor Korolchuk, Newcastle University
  1. FKBP5/FKBP51 on weight watch: central FKBP5 links regulatory WIPI protein networks to autophagy and metabolic control.
  2. Secretory autophagy maintains proteostasis upon lysosome inhibition.
  3. Mapping the proximity of interactome of ATG9A reveals unexpected dynamics of ULK1 complex proteins.
  4. Acetylation in the regulation of autophagy.
  5. ER remodeling via ER-phagy.
  6. Enhanced autophagy and NFE2L2/NRF2 pathway activation in SPOP mutation-driven prostate cancer.
  7. Impaired degradation of YAP1 and IL6ST by chaperone-mediated autophagy promotes proliferation and migration of normal and hepatocellular carcinoma cells.
  8. Autophagy in cancer cell remodeling and quality control.
  9. Cholesterol suppresses GOLM1-dependent selective autophagy of RTKs in hepatocellular carcinoma.
  10. Cylindromatosis drives synapse pruning and weakening by promoting macroautophagy through Akt-mTOR signaling.
  11. The ATG5 interactome links clathrin-mediated vesicular trafficking with the autophagosome assembly machinery.
  12. Post-Translational Modifications of ATG4B in the Regulation of Autophagy.
  13. SGPL1 stimulates VPS39 recruitment to the mitochondria in MICU1 deficient cells.
  14. Cancer cells adapt FAM134B/BiP mediated ER-phagy to survive hypoxic stress.
  15. Only solid waste, please!
  16. Mitochondrial ROS Produced in Human Colon Carcinoma Associated with Cell Survival via Autophagy.
  17. mTORC1 Regulates High Levels of Protein Synthesis in Retinal Ganglion Cells of Adult Mice.
  18. Lysosomes Dysfunction Causes Mitophagy Impairment in PBMCs of Sporadic ALS Patients.
  19. ATG4A regulates human erythroid maturation and mitochondrial clearance.
  20. Dimerization-dependent membrane tethering by Atg23 is essential for yeast autophagy.
  21. The Two-Faced Role of Autophagy in Endometrial Cancer.
  22. You shall not pass! Protective role of autophagic machinery in response to plasma membrane damage triggered by Candida albicans invasion.
  23. Autophagy Signaling by Neural-Induced Human Adipose Tissue-Derived Stem Cell-Conditioned Medium during Rotenone-Induced Toxicity in SH-SY5Y Cells.
  24. Distinct and additive effects of calorie restriction and rapamycin in aging skeletal muscle.
  25. The role of autophagy in the metabolism and differentiation of stem cells.
  26. Autophagy profiling in single cells with open source CellProfiler-based image analysis.
  27. Parkin as a Molecular Bridge Linking Alzheimer's and Parkinson's Diseases?
  28. Cancer cell's internal and external warriors: Autophagosomes and exosomes.
  29. Steady-State Levels of Miro1 Linked to Phosphorylation at Serine 156 and Mitochondrial Respiration in Dopaminergic Neurons.
  30. Translational control by heme-regulated elF2α kinase during erythropoiesis.
  31. Poly(I:C) attenuates myocardial ischemia/reperfusion injury by restoring autophagic function.
  32. mTOR deletion ameliorates CD4 + T cell apoptosis during sepsis by improving autophagosome-lysosome fusion.
  33. SLC38A10 Regulate Glutamate Homeostasis and Modulate the AKT/TSC2/mTOR Pathway in Mouse Primary Cortex Cells.
  34. Calorie Restriction-Regulated Molecular Pathways and Its Impact on Various Age Groups: An Overview.
  35. Autophagy in muscle regeneration: potential therapies for myopathies.
  36. Mitochondria and Other Organelles in Neural Development and Their Potential as Therapeutic Targets in Neurodegenerative Diseases.
  37. SOD1 mediates lysosome-to-mitochondria communication and its dysregulation by amyloid-β oligomers.
  38. The Autophagy Process in Cervical Carcinogenesis: Role of Non-Coding-RNAs, Molecular Mechanisms, and Therapeutic Targets.
  39. Autophagy in Rheumatic Diseases: Role in the Pathogenesis and Therapeutic Approaches.
  40. Detection of Autophagy-Related Structures in Fruiting Bodies of Edible Mushroom, Pleurotus ostreatus.
  41. Signaling Pathways That Mediate Alveolar Macrophage Activation by Surfactant Protein A and IL-4.
  42. SnapShot: Organelle degradation.
  43. Downregulation of CASTOR1 Inhibits Heat-Stress-Induced Apoptosis and Promotes Casein and Lipid Synthesis in Mammary Epithelial Cells.
  44. Protein Kinase A Regulates Autophagy-Associated Proteins Impacting Growth and Virulence of Aspergillus fumigatus.
  45. Acetaldehyde Induces Cytotoxicity via Triggering Mitochondrial Dysfunction and Overactive Mitophagy.
  46. The Complex Roles of Autophagy in Plant Immunity.
  47. TORC1 Signaling Controls the Stability and Function of α-Arrestins Aly1 and Aly2.
  48. UBR5 is a novel regulator of WNK1 stability.
  49. Mitochondrion-Localized SND1 Promotes Mitophagy and Liver Cancer Progression Through PGAM5.
  50. Cannabidiol induces autophagy and improves neuronal health associated with SIRT1 mediated longevity.
  51. FGF21 alleviates pulmonary hypertension by inhibiting mTORC1/EIF4EBP1 pathway via H19.
  52. The goodies of chelated fat: iron-regulated lipid droplet biogenesis precedes and preserves mitophagy.
  53. Flubendazole induces mitochondrial dysfunction and DRP1-mediated mitophagy by targeting EVA1A in breast cancer.
  54. The autophagy-related proteins FvAtg4 and FvAtg8 are involved in virulence and fumonisin biosynthesis in Fusarium verticillioides.
  55. High cholesterol and 27-hydroxycholesterol contribute to phosphorylation of tau protein by impairing autophagy causing learning and memory impairment in C57BL/6J mice.
  56. Implication of autophagy in the antifibrogenic effect of Rilpivirine: when more is less.
  57. Are stressed out goblets cells to blame for functional dyspepsia symptoms?
  58. Quality Control: Maintaining molecular order and preventing cellular chaos.
  59. Pan-mTOR inhibitors sensitize the senolytic activity of Navitoclax via mTORC2 inhibition-mediated apoptotic signaling.
  60. USP13: Multiple Functions and Target Inhibition.
  61. Cellular abundance of sodium phosphate co-transporter SLC20A1/PiT1 and phosphate uptake are controlled post-transcriptionally by ESCRT.
  62. Cisplatin resistance can be curtailed by blunting Bnip3-mediated mitochondrial autophagy.
  63. Phospholipids alter activity and stability of mitochondrial membrane-bound ubiquitin ligase MARCH5.
  64. DnaJC7 in Amyotrophic Lateral Sclerosis.
  65. Programming axonal mitochondrial maintenance and bioenergetics in neurodegeneration and regeneration.
  66. Tau acetylation reduces its autophagic degradation and is a targetable pathway for human tauopathies (comment on https://doi.org/10.1002/bies.201600224).
  67. Squalene-Based Nano-Assemblies Improve the Pro-Autophagic Activity of Trehalose.
  68. Editorial: The Regulation of Autophagy Activity in Pathogenic Microorganism Infection.
  69. AAV-CRISPR/Cas9 Gene Editing Preserves Long-Term Vision in the P23H Rat Model of Autosomal Dominant Retinitis Pigmentosa.
  70. Vitamin D Attenuates Airway Inflammation in Asthmatic Guinea Pigs Using Mammalian Target of Rapamycin-Mediated Autophagy.
  71. Apoptosis and autophagy coordinately shape vector tolerance to arbovirus infection.
  72. Jian Pi Tiao Gan Yin alleviates obesity phenotypes through mTORC1/SREBP1 signaling in vitro and in vivo.
  73. Restoration of ER proteostasis attenuates remote apoptotic cell death after spinal cord injury by reducing autophagosome overload.
  74. A hundred spotlights on microbiology: how microorganisms shape our lives.
  75. Exploratory Analysis of Selected Components of the mTOR Pathway Reveals Potentially Crucial Associations with Childhood Malnutrition.
  1. Autophagy. 2022 Apr 19. 1-3
    FKBP5/FKBP51 on weight watch: central FKBP5 links regulatory WIPI protein networks to autophagy and metabolic control.
    Thomas Bajaj, Alexander S Häusl, Mathias V Schmidt, Nils C Gassen.
      Stress and changes in energy stores are perceived by hormone- and nutrient-sensing nuclei of the hypothalamus, which orchestrate an adaptive physiological body response to maintain homeostasis. Macroautophagy/autophagy is a fundamental lysosomal degradation system contributing to preservation of proteome balance and metabolic homeostasis. Its dysregulation is linked to diverse human pathologies, including neuropsychiatric and metabolic disorders. Autophagy is coordinated by cellular nutrient sensors, including AMPK and MTORC1 that interact with WIPI proteins. Studies suggest that WDR45/WIPI4 interacts with the stress-sensitive co-chaperone FKBP5/FKBP51, which has emerged as a key autophagy scaffold. However, the impact of FKBP5 on autophagy signaling in response to metabolic challenges, such as a high-fat diet, is elusive. Therefore, we manipulated FKBP5 in the mediobasal hypothalamus (MBH) and studied autophagy signaling and protein interactions in their physiological context. We identified FKBP5 as a scaffold of the STK11/LKB1-AMPK complex with WDR45/WIPI4 and TSC2 with WDR45B/WIPI3 in response to metabolic challenges, positioning FKBP5 in major nutrient-sensing and autophagy-regulating networks. Intriguingly, we could demonstrate that FKBP5 deletion in the MBH strongly induces obesity, whereas its overexpression protects against high-fat diet-induced obesity. Our findings suggest a crucial regulatory and adaptive function of FKBP5-regulated autophagy within the MBH in response to metabolic challenges.Abbreviations: AKT: thymoma viral proto-oncogene; AMPK: AMP-activated protein kinase; BECN1: beclin 1, autophagy related; eWAT: epididymal white adipose tissue; FKBP5/FKBP51: FK506 binding protein 5; KO, knockout; MBH, mediobasal hypothalamus; MTORC1, mechanistic target of rapamycin kinase complex 1; p: phosphorylated; PHLPP: PH domain and leucine rich repeat protein phosphatase; RPS6KB/p70S6K: ribosomal protein S6 kinase; SKP2: S-phase kinase-associated protein 2; SM: soleus muscle; SQSTM1/p62, sequestosome 1; STK11/LKB1: serine/threonine kinase 11; TSC: TSC complex; ULK1: unc-51 like kinase 1; WIPI: WD repeat domain, phosphoinositide interacting; WT: wild type.
    Keywords:  AMPK; FKBP5/FKBP51; WIPI; autophagy; metabolic stress
    DOI:  https://doi.org/10.1080/15548627.2022.2063006
  2. J Cell Biol. 2022 Jun 06. pii: e202110151. [Epub ahead of print]221(6):
    Secretory autophagy maintains proteostasis upon lysosome inhibition.
    Tina A Solvik, Tan A Nguyen, Yu-Hsiu Tony Lin, Timothy Marsh, Eric J Huang, Arun P Wiita, Jayanta Debnath, Andrew M Leidal.
      The endolysosome system plays central roles in both autophagic degradation and secretory pathways, including the release of extracellular vesicles and particles (EVPs). Although previous work reveals important interconnections between autophagy and EVP-mediated secretion, our understanding of these secretory events during endolysosome inhibition remains incomplete. Here, we delineate a secretory autophagy pathway upregulated in response to endolysosomal inhibition, which mediates EVP-associated release of autophagic cargo receptors, including p62/SQSTM1. This secretion is highly regulated and dependent on multiple ATGs required for autophagosome formation, as well as the small GTPase Rab27a. Furthermore, disrupting autophagosome maturation, either via genetic inhibition of autophagosome-to-autolysosome fusion or expression of SARS-CoV-2 ORF3a, is sufficient to induce EVP secretion of autophagy cargo receptors. Finally, ATG-dependent EVP secretion buffers against the intracellular accumulation of autophagy cargo receptors when classical autophagic degradation is impaired. Thus, we propose secretory autophagy via EVPs functions as an alternate route to clear sequestered material and maintain proteostasis during endolysosomal dysfunction or impaired autophagosome maturation.
    DOI:  https://doi.org/10.1083/jcb.202110151
  3. Autophagy. 2022 Apr 20. 1-2
    Mapping the proximity of interactome of ATG9A reveals unexpected dynamics of ULK1 complex proteins.
    Ashari R Kannangara, Joshua L Andersen.
      ATG9A is essential for macroautophagy/autophagy and considered to be one of the earliest ATG (autophagy related) proteins recruited to sites of autophagosome biogenesis. Recent data suggest ATG9A vesicles may even form the lipid seed of the autophagosome. However, ATG9A regulation is still poorly understood, which is likely at least partly due to challenges inherent to studying an intracellular transmembrane protein with no apparent enzymatic activity. To help overcome these challenges, we used BioID and quantitative LC-MS/MS to map the proximity interactome of ATG9A, which included entire protein complexes involved in protein trafficking, and proteins implicated in autophagy but previously lacking any physical link to core autophagy machinery. We also unexpectedly found an ATG9A interaction with an ULK1-independent ATG13-ATG101 dimer that promotes autophagy in fed cells.
    Keywords:  ATG13; ATG9A; ULK1; basal autophagy. BioID; proteomics
    DOI:  https://doi.org/10.1080/15548627.2022.2062953
  4. Autophagy. 2022 Apr 18. 1-9
    Acetylation in the regulation of autophagy.
    Yinfeng Xu, Wei Wan.
      Post-translational modifications, such as phosphorylation, ubiquitination and acetylation, play crucial roles in the regulation of autophagy. Acetylation has emerged as an important regulatory mechanism for autophagy. Acetylation regulates autophagy initiation and autophagosome formation by targeting core components of the ULK1 complex, the BECN1-PIK3C3 complex, and the LC3 lipidation system. Recent studies have shown that acetylation occurs on the key proteins participating in autophagic cargo assembly and autophagosome-lysosome fusion, such as SQSTM1/p62 and STX17. In addition, acetylation controls autophagy at the transcriptional level by targeting histones and the transcription factor TFEB. Here, we review the current knowledge on acetylation of autophagy proteins and their regulations and functions in the autophagy pathway with focus on recent findings.Abbreviations : ACAT1: acetyl-CoA acetyltransferase 1; ACSS2: acyl-CoA synthetase short chain family member 2; AMPK: AMP-activated protein kinase; ATG: autophagy-related; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; CCAR2/DBC1: cell cycle and apoptosis regulator 2; BECN1: beclin 1; CMA: chaperone-mediated autophagy; CREBBP/CBP: CREB binding protein; EP300/p300: E1A binding protein p300; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GSK3: glycogen synthase kinase 3; HDAC6: histone deacetylase 6; HSPA8/HSC70: heat shock protein family A (Hsp70) member 8; KAT2A/GCN5: lysine acetyltransferase 2A; KAT2B/PCAF: lysine acetyltransferase 2B; KAT5/TIP60: lysine acetyltransferase 5; KAT8/MOF: lysine acetyltransferase 8; LAMP2A: lysosomal associated membrane protein 2A; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; NBR1: NBR1 autophagy cargo receptor; OPTN: optineurin; PD: Parkinson disease; PE: phosphatidylethanolamine; PIK3C3/VPS34: phosphatidylinositol 3-kinase catalytic subunit type 3; PKM2: pyruvate kinase M1/2; PtdIns3P: phosphatidylinositol-3-phosphate; PTM: post-translational modification; RB1CC1/FIP200: RB1 inducible coiled-coil 1; RUBCN/Rubicon: rubicon autophagy regulator; RUBCNL/Pacer: rubicon like autophagy enhancer; SIRT1: sirtuin 1; SNAP29: synaptosome associated protein 29; SNARE: soluble N-ethylamide-sensitive factor attachment protein receptor; SQSTM1/p62: sequestosome 1; STX17: syntaxin 17; TFEB: transcription factor EB; TP53/p53: tumor protein p53; TP53INP2/DOR: tumor protein p53 inducible nuclear protein 2; UBA: ubiquitin-associated; ULK1: unc-51 like autophagy activating kinase 1; VAMP8: vesicle associated membrane protein 8; WIPI2: WD repeat domain, phosphoinositide interacting 2.
    Keywords:  Acetylation; acetyltransferase; autophagy; deacetylase; post-translational modification
    DOI:  https://doi.org/10.1080/15548627.2022.2062112
  5. Mol Cell. 2022 Apr 21. pii: S1097-2765(22)00156-3. [Epub ahead of print]82(8): 1492-1500
    ER remodeling via ER-phagy.
    Andrea Gubas, Ivan Dikic.
      The endoplasmic reticulum (ER) is a hotspot for many essential cellular functions. The ER membrane is highly dynamic, which affects many cellular processes that take place within the ER. One such process is ER-phagy, a selective degradation of ER fragments (including membranes and luminal content), which serves to preserve the size of ER while adapting its morphology under basal and stress conditions. In order to be degraded, the ER undergoes selective fragmentation facilitated by specialized ER-shaping proteins that also act as ER-phagy receptors. Their ability to sense and induce membrane curvature, as well as to bridge the ER with autophagy machinery, allows for a successful ER fragmentation and delivery of these fragments to the lysosome for degradation and recycling. In this review, we provide insights into ER-phagy from the perspective of membrane remodeling. We highlight the importance of ER membrane dynamics during ER-phagy and emphasize how its dysregulation reflects on human physiology and pathology.
    DOI:  https://doi.org/10.1016/j.molcel.2022.02.018
  6. Autophagy. 2022 Apr 19. 1-3
    Enhanced autophagy and NFE2L2/NRF2 pathway activation in SPOP mutation-driven prostate cancer.
    Kun Gao, Qing Shi, Yajuan Liu, Chenji Wang.
      SQSTM1/p62 is a selective macroautophagy/autophagy receptor that drives ubiquitinated cargos toward the lysosome for degradation, and also a stress-induced scaffold protein that helps cells to cope with oxidative stress through sequestrating KEAP1 and subsequent activation of the NFE2L2/NRF2 antioxidant pathway. Accumulating evidence implicates SQSTM1 dysregulation in the induction of multiple oncogenic transformations in vivo. SPOP (speckle type BTB/POZ protein), an E3 ubiquitin ligase adaptor, is the most frequently mutated gene in prostate cancer (Pca), but the molecular mechanisms underlying how SPOP mutations contribute to PCa tumorigenesis are still largely unknown. In a recent study, we describe a new role for SPOP as a negative regulator of autophagy and NFE2L2 pathway activation. SPOP binds and induces the non-degradative ubiquitination of SQSTM1 at Lys420. This post-translational modification decreases SQSTM1 body formation, liquid phase condensation, dimerization, and ubiquitin-binding capacity, thereby suppressing SQSTM1-dependent autophagy, KEAP1 sequestration, and NFE2L2 activation. Notably, PCa-associated SPOP mutants lose the capacity to ubiquitinate SQSTM1 and instead enhance autophagy and the antioxidant response in a dominant-negative manner. Thus, our findings indicate the critical roles of autophagy and NFE2L2 pathway activation in PCa tumorigenesis by oncogenic SPOP mutations.
    Keywords:  Gene mutation; KEAP1; SPOP; SQSTM1; oxidative stress; phase separation; prostate cancer; ubiquitination
    DOI:  https://doi.org/10.1080/15548627.2022.2062873
  7. Autophagy. 2022 Apr 18. 1-11
    Impaired degradation of YAP1 and IL6ST by chaperone-mediated autophagy promotes proliferation and migration of normal and hepatocellular carcinoma cells.
    Enrico Desideri, Serena Castelli, Coralie Dorard, Stefanie Toifl, Gian Luca Grazi, Maria Rosa Ciriolo, Manuela Baccarini.
      Impaired degradation of the transcriptional coactivator YAP1 and IL6ST (interleukin 6 cytokine family signal transducer), two proteins deregulated in liver cancer, has been shown to promote tumor growth. Here, we demonstrate that YAP1 and IL6ST are novel substrates of chaperone-mediated autophagy (CMA) in human hepatocellular carcinoma (HCC) and hepatocyte cell lines. Knockdown of the lysosomal CMA receptor LAMP2A increases protein levels of YAP1 and IL6ST, without changes in mRNA expression. Additionally, both proteins show KFERQ-dependent binding to the CMA chaperone HSPA8 and accumulate into isolated lysosomes after stimulation of CMA by prolonged starvation. We further show that LAMP2A downregulation promotes the proliferation and migration in HCC cells and a human hepatocyte cell line, and that it does so in a YAP1- and IL6ST-dependent manner. Finally, LAMP2A expression is downregulated, and YAP1 and IL6ST expression is upregulated, in human HCC biopsies. Taken together, our work reveals a novel mechanism that controls the turnover of two cancer-relevant proteins and suggests a tumor suppressor function of CMA in the liver, advocating for the exploitation of CMA activity for diagnostic and therapeutic purposes.Abbreviations: ACTB: actin beta; ATG5: autophagy related 5; ATG7: autophagy related 7; CMA: chaperone-mediated autophagy; eMI: endosomal microautophagy; HCC: hepatocellular carcinoma; HSPA8: heat shock protein family A (Hsp70) member 8; IL6ST: interleukin 6 cytokine family signal transducer; JAK: Janus kinase; LAMP1: lysosomal associated membrane protein 1; LAMP2A: lysosomal associated membrane protein 2A; MAPK8: mitogen-activated protein kinase 8; P6: pyridine 6; SQSTM1: sequestosome 1; TUBA: tubulin alpha; VDAC1: voltage dependent anion channel 1; VP: verteporfin; YAP1: Yes1 associated transcriptional regulator.
    Keywords:  Chaperone-mediated autophagy; IL6ST; KFERQ motif; LAMP2A; YAP1; hepatocellular carcinoma; migration; proliferation; protein degradation
    DOI:  https://doi.org/10.1080/15548627.2022.2063004
  8. Mol Cell. 2022 Apr 21. pii: S1097-2765(22)00261-1. [Epub ahead of print]82(8): 1514-1527
    Autophagy in cancer cell remodeling and quality control.
    Grace A Hernandez, Rushika M Perera.
      As one of the two highly conserved cellular degradation systems, autophagy plays a critical role in regulation of protein, lipid, and organelle quality control and cellular homeostasis. This evolutionarily conserved pathway singles out intracellular substrates for elimination via encapsulation within a double-membrane vesicle and delivery to the lysosome for degradation. Multiple cancers disrupt normal regulation of autophagy and hijack its degradative ability to remodel their proteome, reprogram their metabolism, and adapt to environmental challenges, making the autophagy-lysosome system a prime target for anti-cancer interventions. Here, we discuss the roles of autophagy in tumor progression, including cancer-specific mechanisms of autophagy regulation and the contribution of tumor and host autophagy in metabolic regulation, immune evasion, and malignancy. We further discuss emerging proteomics-based approaches for systematic profiling of autophagosome-lysosome composition and contents. Together, these approaches are uncovering new features and functions of autophagy, leading to more effective strategies for targeting this pathway in cancer.
    Keywords:  autophagy; cancer; lysosome; quality control; remodeling
    DOI:  https://doi.org/10.1016/j.molcel.2022.03.023
  9. Cell Rep. 2022 Apr 19. pii: S2211-1247(22)00473-9. [Epub ahead of print]39(3): 110712
    Cholesterol suppresses GOLM1-dependent selective autophagy of RTKs in hepatocellular carcinoma.
    Wei-Qing Shao, Wen-Wei Zhu, Meng-Jun Luo, Ming-Hao Fan, Qin Li, Sheng-Hao Wang, Zhi-Fei Lin, Jing Zhao, Yan Zheng, Qiong-Zhu Dong, Lu Lu, Hu-Liang Jia, Ju-Bo Zhang, Ming Lu, Jin-Hong Chen, Lun-Xiu Qin.
      Aberrant activation of receptor tyrosine kinases (RTKs) and the subsequent metabolic reprogramming play critical roles in cancer progression. Our previous study has shown that Golgi membrane protein 1 (GOLM1) promotes hepatocellular carcinoma (HCC) metastasis by enhancing the recycling of RTKs. However, how this RTK recycling process is regulated and coupled with RTK degradation remains poorly defined. Here, we demonstrate that cholesterol suppresses the autophagic degradation of RTKs in a GOLM1-dependent manner. Further mechanistic studies reveal that GOLM1 mediates the selective autophagy of RTKs by interacting with LC3 through an LC3-interacting region (LIR), which is regulated by a cholesterol-mTORC1 axis. Lowering cholesterol by statins improves the efficacy of multiple tyrosine kinase inhibitors (TKIs) in vivo. Our findings indicate that cholesterol serves as a signal to switch GOLM1-RTK degradation to GOLM1-RTK recycling and suggest that lowering cholesterol by statin may be a promising combination strategy to improve the TKI efficiency in HCC.
    Keywords:  CP: Cancer; CP: Cell biology; cholesterol metabolism; liver cancer; lysosomal degradation; statin; tyrosine kinase inhibitor
    DOI:  https://doi.org/10.1016/j.celrep.2022.110712
  10. Mol Psychiatry. 2022 Apr 21.
    Cylindromatosis drives synapse pruning and weakening by promoting macroautophagy through Akt-mTOR signaling.
    Alexis S Zajicek, Hongyu Ruan, Huihui Dai, Mary C Skolfield, Hannah L Phillips, Wendi J Burnette, Behnam Javidfar, Shao-Cong Sun, Schahram Akbarian, Wei-Dong Yao.
      The lysine-63 deubiquitinase cylindromatosis (CYLD) is long recognized as a tumor suppressor in immunity and inflammation, and its loss-of-function mutations lead to familial cylindromatosis. However, recent studies reveal that CYLD is enriched in mammalian brain postsynaptic densities, and a gain-of-function mutation causes frontotemporal dementia (FTD), suggesting critical roles at excitatory synapses. Here we report that CYLD drives synapse elimination and weakening by acting on the Akt-mTOR-autophagy axis. Mice lacking CYLD display abnormal sociability, anxiety- and depression-like behaviors, and cognitive inflexibility. These behavioral impairments are accompanied by excessive synapse numbers, increased postsynaptic efficacy, augmented synaptic summation, and impaired NMDA receptor-dependent hippocampal long-term depression (LTD). Exogenous expression of CYLD results in removal of established dendritic spines from mature neurons in a deubiquitinase activity-dependent manner. In search of underlying molecular mechanisms, we find that CYLD knockout mice display marked overactivation of Akt and mTOR and reduced autophagic flux, and conversely, CYLD overexpression potently suppresses Akt and mTOR activity and promotes autophagy. Consequently, abrogating the Akt-mTOR-autophagy signaling pathway abolishes CYLD-induced spine loss, whereas enhancing autophagy in vivo by the mTOR inhibitor rapamycin rescues the synaptic pruning and LTD deficits in mutant mice. Our findings establish CYLD, via Akt-mTOR signaling, as a synaptic autophagy activator that exerts critical modulations on synapse maintenance, function, and plasticity.
    DOI:  https://doi.org/10.1038/s41380-022-01571-1
  11. Autophagy Rep. 2022 ;1(1): 88-118
    The ATG5 interactome links clathrin-mediated vesicular trafficking with the autophagosome assembly machinery.
    Kiren Baines, Kazuaki Yoshioka, Yoh Takuwa, Jon D Lane.
      Autophagosome formation involves the sequential actions of conserved ATG proteins to coordinate the lipidation of the ubiquitin-like modifier Atg8-family proteins at the nascent phagophore membrane. Although the molecular steps driving this process are well understood, the source of membranes for the expanding phagophore and their mode of delivery are only now beginning to be revealed. Here, we have used quantitative SILAC-based proteomics to identify proteins that associate with the ATG12-ATG5 conjugate, a crucial player during Atg8-family protein lipidation. Our datasets reveal a strong enrichment of regulators of clathrin-mediated vesicular trafficking, including clathrin heavy and light chains, and several clathrin adaptors. Also identified were PIK3C2A (a phosphoinositide 3-kinase involved in clathrin-mediated endocytosis) and HIP1R (a component of clathrin vesicles), and the absence of either of these proteins alters autophagic flux in cell-based starvation assays. To determine whether the ATG12-ATG5 conjugate reciprocally influences trafficking within the endocytic compartment, we captured the cell surface proteomes of autophagy-competent and autophagy-incompetent mouse embryonic fibroblasts under fed and starved conditions. We report changes in the relative proportions of individual cell surface proteins and show that cell surface levels of the SLC7A5-SLC3A2 amino acid transporter are influenced by autophagy capability. Our data provide evidence for direct regulatory coupling between the ATG12-ATG5 conjugate and the clathrin membrane trafficking system and suggest candidate membrane proteins whose trafficking within the cell may be modulated by the autophagy machinery. Abbreviations: ATG, autophagy related; BafA1, bafilomycin A1; GFP, green fluorescent protein; HIP1R, huntingtin interacting protein 1 related; MEF, mouse embryo fibroblast; PIK3C2A, phosphatidylinositol-4-phosphate 3-kinase catalytic subunit type 2 alpha; SILAC, stable isotope labelling with amino acids in culture; SQSTM1, sequestosome 1; STRING, search tool for the retrieval of interacting genes/proteins.
    Keywords:  ATG12; ATG5; HIP1R; PIK3C2A; autophagy; clathrin; endocytosis; proteomics
    DOI:  https://doi.org/10.1080/27694127.2022.2042054
  12. Cells. 2022 Apr 13. pii: 1330. [Epub ahead of print]11(8):
    Post-Translational Modifications of ATG4B in the Regulation of Autophagy.
    Na Yeon Park, Doo Sin Jo, Dong-Hyung Cho.
      Autophagy plays a key role in eliminating and recycling cellular components in response to stress, including starvation. Dysregulation of autophagy is observed in various diseases, including neurodegenerative diseases, cancer, and diabetes. Autophagy is tightly regulated by autophagy-related (ATG) proteins. Autophagy-related 4 (ATG4) is the sole cysteine protease, and four homologs (ATG4A-D) have been identified in mammals. These proteins have two domains: catalytic and short fingers. ATG4 facilitates autophagy by promoting autophagosome maturation through reversible lipidation and delipidation of seven autophagy-related 8 (ATG8) homologs, including microtubule-associated protein 1-light chain 3 (LC3) and GABA type A receptor-associated protein (GABARAP). Each ATG4 homolog shows a preference for a specific ATG8 homolog. Post-translational modifications of ATG4, including phosphorylation/dephosphorylation, O-GlcNAcylation, oxidation, S-nitrosylation, ubiquitination, and proteolytic cleavage, regulate its activity and ATG8 processing, thus modulating its autophagic activity. We reviewed recent advances in our understanding of the effect of post-translational modification on the regulation, activity, and function of ATG4, the main protease that controls autophagy.
    Keywords:  ATG4; autophagy; post-translational modification
    DOI:  https://doi.org/10.3390/cells11081330
  13. Mol Metab. 2022 Apr 19. pii: S2212-8778(22)00072-2. [Epub ahead of print] 101503
    SGPL1 stimulates VPS39 recruitment to the mitochondria in MICU1 deficient cells.
    Joshua Jackson, Lena Wischhof, Enzo Scifo, Anna Pellizzer, Yiru Wang, Antonia Piazzesi, Debora Gentile, Sana Siddig, Miriam Stork, Chris E Hopkins, Kristian Händler, Joachim Weis, Andreas Roos, Joachim L Schultze, Pierluigi Nicotera, Dan Ehninger, Daniele Bano.
      OBJECTIVE: Mitochondrial "retrograde" signaling may stimulate organelle biogenesis as a compensatory adaptation to aberrant activity of the oxidative phosphorylation (OXPHOS) system. To maintain energy-consuming processes in OXPHOS deficient cells, alternative metabolic pathways are functionally coupled to the degradation, recycling and redistribution of biomolecules across distinct intracellular compartments. While transcriptional regulation of mitochondrial network expansion has been the focus of many studies, the molecular mechanisms promoting mitochondrial maintenance in energy-deprived cells remain poorly investigated.METHODS: We performed transcriptomics, quantitative proteomics and lifespan assays to identify pathways that are mechanistically linked to mitochondrial network expansion and homeostasis in Caenorhabditis elegans lacking the mitochondrial calcium uptake protein 1 (MICU-1/MICU1). To support our findings, we carried out biochemical and image analyses in mammalian cells and mouse-derived tissues.
    RESULTS: We report that micu-1(null) mutations impair the OXPHOS system and promote C. elegans longevity through a transcriptional program that is independent of the mitochondrial calcium uniporter MCU-1/MCU and the essential MCU regulator EMRE-1/EMRE. We identify sphingosine phosphate lyase SPL-1/SGPL1 and the ATFS-1-target HOPS complex subunit VPS-39/VPS39 as critical lifespan modulators of micu-1(null) mutant animals. Cross-species investigation indicates that SGPL1 upregulation stimulates VPS39 recruitment to the mitochondria, thereby enhancing mitochondria-lysosome contacts. Consistently, VPS39 downregulation compromises mitochondrial network maintenance and basal autophagic flux in MICU1 deficient cells. In mouse-derived muscles, we show that VPS39 recruitment to the mitochondria may represent a common signature associated with altered OXPHOS system.
    CONCLUSIONS: Our findings reveal a previously unrecognized SGPL1/VPS39 axis that stimulates intracellular organelle interactions and sustains autophagy and mitochondrial homeostasis in OXPHOS deficient cells.
    Keywords:  Caenorhabditis elegans; MICU1; VPS39; autophagy; longevity; mitochondria; sphingosine signaling
    DOI:  https://doi.org/10.1016/j.molmet.2022.101503
  14. Cell Death Dis. 2022 Apr 18. 13(4): 357
    Cancer cells adapt FAM134B/BiP mediated ER-phagy to survive hypoxic stress.
    Sandhya Chipurupalli, Raja Ganesan, Giulia Martini, Luigi Mele, Alessio Reggio, Marianna Esposito, Elango Kannan, Vigneshwaran Namasivayam, Paolo Grumati, Vincenzo Desiderio, Nirmal Robinson.
      In the tumor microenvironment, cancer cells experience hypoxia resulting in the accumulation of misfolded/unfolded proteins largely in the endoplasmic reticulum (ER). Consequently, ER proteotoxicity elicits unfolded protein response (UPR) as an adaptive mechanism to resolve ER stress. In addition to canonical UPR, proteotoxicity also stimulates the selective, autophagy-dependent, removal of discrete ER domains loaded with misfolded proteins to further alleviate ER stress. These mechanisms can favor cancer cell growth, metastasis, and long-term survival. Our investigations reveal that during hypoxia-induced ER stress, the ER-phagy receptor FAM134B targets damaged portions of ER into autophagosomes to restore ER homeostasis in cancer cells. Loss of FAM134B in breast cancer cells results in increased ER stress and reduced cell proliferation. Mechanistically, upon sensing hypoxia-induced proteotoxic stress, the ER chaperone BiP forms a complex with FAM134B and promotes ER-phagy. To prove the translational implication of our mechanistic findings, we identified vitexin as a pharmacological agent that disrupts FAM134B-BiP complex, inhibits ER-phagy, and potently suppresses breast cancer progression in vivo.
    DOI:  https://doi.org/10.1038/s41419-022-04813-w
  15. Mol Cell. 2022 Apr 21. pii: S1097-2765(22)00314-8. [Epub ahead of print]82(8): 1408-1410
    Only solid waste, please!
    Dorothee Dormann, Christian Behrends.
      To elucidate the mechanism driving selective autophagy of protein aggregates, or "aggrephagy," Ma et al. (2022) identify chaperonin TRiC subunit CCT2 as a receptor that specifically promotes the clearance of solid aggregates, but not liquid-like condensates, in a ubiquitin-independent manner.
    DOI:  https://doi.org/10.1016/j.molcel.2022.04.003
  16. Cancers (Basel). 2022 Apr 08. pii: 1883. [Epub ahead of print]14(8):
    Mitochondrial ROS Produced in Human Colon Carcinoma Associated with Cell Survival via Autophagy.
    Eun Ji Gwak, Dasol Kim, Hui-Yun Hwang, Ho Jeong Kwon.
      Human colon carcinomas, including HCT116 cells, often exhibit high autophagic flux under nutrient deprivation or hypoxic conditions. Mitochondrial ROS (mROS) is known as a 'molecular switch' for regulating the autophagic pathway, which is critical for directing cancer cell survival or death. In early tumorigenesis, autophagy plays important roles in maintaining cellular homeostasis and contributes to tumor growth. However, the relationships between mROS and the autophagic capacities of HCT116 cells are poorly understood. Ubiquinol cytochrome c reductase binding protein (UQCRB) has been reported as a biomarker of colorectal cancer, but its role in tumor growth has not been clarified. Here, we showed that UQCRB is overexpressed in HCT116 cells compared to CCD18co cells, a normal colon fibroblast cell line. Pharmacological inhibition of UQCRB reduced mROS levels, autophagic flux, and the growth of HCT116 tumors in a xenograft mouse model. We further investigated mutant UQCRB-overexpressing cell lines to identify functional links in UQCRB-mROS-autophagy. Notably, an increasing level of mROS caused by UQCRB overexpression released Ca2+ by the activation of lysosomal transient receptor potential mucolipin 1 (TRPML1) channels. This activation induced transcription factor EB (TFEB) nuclear translocation and lysosome biogenesis, leading to autophagy flux. Collectively, our study showed that increasing levels of mROS caused by the overexpression of UQCRB in human colon carcinoma HCT116 cells could be linked to autophagy for cell survival.
    Keywords:  UQCRB; autophagy; colorectal cancer; lysosome; mROS
    DOI:  https://doi.org/10.3390/cancers14081883
  17. J Biol Chem. 2022 Apr 18. pii: S0021-9258(22)00384-2. [Epub ahead of print] 101944
    mTORC1 Regulates High Levels of Protein Synthesis in Retinal Ganglion Cells of Adult Mice.
    Patrice E Fort, Mandy K Losiewicz, Lynda Elghazi, Dejuan Kong, Corentin Cras-Méneur, Diane C Fingar, Scot R Kimball, Raju V S Rajala, Alexander J Smith, Robin R Ali, Steven F Abcouwer, Thomas W Gardner.
      Mechanistic target of rapamycin (mTOR) and members of mTOR complex 1 (mTORC1), a linchpin of the nutrient sensing and protein synthesis pathways, are present at relatively high levels in the ganglion cell layer (GCL) and retinal ganglion cells (RGC) of rodent and human retinas. However, the role of mTOR complexes in the control of protein synthesis in RGC is unknown. Here we applied the SUnSET method of nascent protein labeling to localize and quantify protein synthesis in the retinas of adult mice. We also used intravitreal injection of an AAV2 vector encoding Cre recombinase in the eyes of mtor- or rptor-floxed mice to conditionally knockout either both mTOR complexes or only mTORC1, respectively, in cells within the GCL. A novel vector encoding an inactive Cre mutant (CreΔC) served as control. We found that retinal protein synthesis was highest in the GCL, particularly in RGC. Negation of both complexes or only mTORC1 significantly reduced protein synthesis in RGC. In addition, loss of mTORC1 function caused a significant reduction in the pan-RGC marker, RNA binding protein with multiple splicing (RBPMS), with little decrease of the total number of cells in the RGC layer, even at 25 weeks after AAV-Cre injection. These findings reveal that mTORC1 signaling is necessary for maintaining the high rate of protein synthesis in RGCs of adult rodents, but it may not be essential to maintain RGC viability. These findings may also be relevant to understanding the pathophysiology of RGC disorders, including glaucoma, diabetic retinopathy, and optic neuropathies.
    Keywords:  eye; gene knockout; mRNA translation; mTOR complex (mTORC); mTORC1; mechanistic target of rapamycin (mTOR); protein synthesis; puromycin; retina; retinal ganglion cell
    DOI:  https://doi.org/10.1016/j.jbc.2022.101944
  18. Cells. 2022 Apr 09. pii: 1272. [Epub ahead of print]11(8):
    Lysosomes Dysfunction Causes Mitophagy Impairment in PBMCs of Sporadic ALS Patients.
    Matteo Bordoni, Orietta Pansarasa, Eveljn Scarian, Riccardo Cristofani, Roberta Leone, Valentina Fantini, Maria Garofalo, Luca Diamanti, Stefano Bernuzzi, Stella Gagliardi, Stephana Carelli, Angelo Poletti, Cristina Cereda.
      Mitochondria alterations are present in tissues derived from patients and animal models, but no data are available for peripheral blood mononuclear cells (PBMCs) of ALS patients. This work aims to investigate mitophagy in PBMCs of sporadic (sALS) patients and how this pathway can be tuned by using small molecules. We found the presence of morphologically atypical mitochondria by TEM and morphological abnormalities by MitoTracker™. We found a decreased number of healthy mitochondria in sALS PBMCs and an impairment of mitophagy with western blot and immunofluorescence. After rapamycin treatment, we found a higher increase in the LC3 marker in sALS PBMCs, while after NH4Cl treatment, we found a lower increase in the LC3 marker. Finally, mTOR-independent autophagy induction with trehalose resulted in a significant decrease in the lysosomes level sALS PBMCs. Our data suggest that the presence of morphologically altered mitochondria and an inefficient turnover of damaged mitochondria in PBMCs of sALS patients rely on the impairment of the mitophagy pathway. We also found that the induction of the mTOR-independent autophagy pathway leads to a decrease in lysosomes level, suggesting a more sensitivity of sALS PBMCs to trehalose. Such evidence suggests that trehalose could represent an effective treatment for ALS patients.
    Keywords:  ALS; PBMCs; autolysosomes; mTOR; mitochondria; trehalose
    DOI:  https://doi.org/10.3390/cells11081272
  19. Blood Adv. 2022 Apr 20. pii: bloodadvances.2021005910. [Epub ahead of print]
    ATG4A regulates human erythroid maturation and mitochondrial clearance.
    Massiel Chavez Stolla, Andreea Reilly, Rochelle Bergantinos, Sintra Stewart, Neele Thom, Courtnee A Clough, Rachel Courtney Wellington, Raisa Stolitenko, Janis L Abkowitz, Sergei Doulatov.
      Autophagy is a self-degradation pathway that is essential for erythropoiesis. During erythroid differentiation, autophagy facilitates the degradation of macromolecules and the programmed clearance of mitochondria. Impaired mitochondrial clearance results in anemia and alters the lifespan of red blood cells in vivo. While several essential autophagy genes contribute to autophagy in erythropoiesis, little is known about erythroid-specific mediators of this pathway. Genetic analysis of primary human erythroid and non-erythroid cells revealed the selective upregulation of the core autophagy gene ATG4A in maturing human erythroid cells. Since the function of ATG4A in erythropoiesis is unknown, we evaluated its role using an ex vivo model of human erythropoiesis. Depletion of ATG4A in primary human hematopoietic stem and progenitor cells selectively impaired erythroid but not myeloid lineage differentiation, resulting in reduced red cell production, delayed terminal differentiation, and impaired enucleation. Loss of ATG4A impaired autophagy and mitochondrial clearance, giving rise to reticulocytes with retained mitochondria and autophagic vesicles. In summary, our study identifies ATG4A as a cell type-specific regulator of autophagy in erythroid development.
    DOI:  https://doi.org/10.1182/bloodadvances.2021005910
  20. Cell Rep. 2022 Apr 19. pii: S2211-1247(22)00460-0. [Epub ahead of print]39(3): 110702
    Dimerization-dependent membrane tethering by Atg23 is essential for yeast autophagy.
    Wayne D Hawkins, Kelsie A Leary, Devika Andhare, Hana Popelka, Daniel J Klionsky, Michael J Ragusa.
      Eukaryotes maintain cellular health through the engulfment and subsequent degradation of intracellular cargo using macroautophagy. The function of Atg23, despite being critical to the efficiency of this process, is unclear due to a lack of biochemical investigations and an absence of any structural information. In this study, we use a combination of in vitro and in vivo methods to show that Atg23 exists primarily as a homodimer, a conformation facilitated by a putative amphipathic helix. We utilize small-angle X-ray scattering to monitor the overall shape of Atg23, revealing that it contains an extended rod-like structure spanning approximately 320 Å. We also demonstrate that Atg23 interacts with membranes directly, primarily through electrostatic interactions, and that these interactions lead to vesicle tethering. Finally, mutation of the hydrophobic face of the putative amphipathic helix completely precludes dimer formation, leading to severely impaired subcellular localization, vesicle tethering, Atg9 binding, and autophagic efficiency.
    Keywords:  CP: Cell biology; autophagy; lysosome; membrane tether; stress; vacuole
    DOI:  https://doi.org/10.1016/j.celrep.2022.110702
  21. Front Cell Dev Biol. 2022 ;10 839416
    The Two-Faced Role of Autophagy in Endometrial Cancer.
    Tomohiko Fukuda, Osamu Wada-Hiraike.
      Autophagy, meaning "self-eating," is a cellular catabolic process that involves lysosomal degradation of cytoplasmic materials. Autophagy contributes to both quality control and energy supply of cells, which are associated with tumorigenesis and tumor development, respectively. Endometrial cancer (EC) is the most common gynecologic cancer, and its incidence is increasing. Although autophagy plays crucial roles in several types of cancer, such as pancreatic ductal adenocarcinoma, its role in EC has not been clearly demonstrated. Activation of the PI3K/AKT/mTOR pathway, which functions to suppress autophagy, is an initial step in type 1 endometrial carcinogenesis, whereas a loss-of-function mutation of TP53, which augments autophagy via p16 induction, is the main cause of type 2 endometrial carcinogenesis. Mutations in autophagy-related genes, including ATG4C, RB1CC1/FIP200, and ULK4, have been reported in EC; thus, an aberrant autophagy mechanism may be involved in endometrial carcinogenesis. Furthermore, the biguanide diabetes drug metformin, treatment with which enhances autophagy via AMPK-mediated mTOR inactivation, has been reported to reduce the risk of EC. These findings suggest that autophagy negatively regulates endometrial carcinogenesis, and autophagy inducers may be useful for chemoprevention of EC. In contrast, autophagy appears to promote EC once it is established. Consistent with this, treatment with chloroquine, an autophagy inhibitor, is reported to attenuate EC cell proliferation. Moreover, chemotherapy-induced autophagy triggers chemoresistance in EC cells. As autophagy has a tumor-promoting function, the combination of chemotherapy and autophagy inhibitors such as chloroquine could be a potent therapeutic option for patients with EC. In conclusion, autophagy plays a dual role in the prevention and treatment of EC. Therefore, targeting autophagy to prevent and treat EC requires diametrically opposed strategies.
    Keywords:  autophagy; chemoprevention; chloroquine; endometrial cancer; metformin
    DOI:  https://doi.org/10.3389/fcell.2022.839416
  22. Autophagy. 2022 Apr 20. 1-2
    You shall not pass! Protective role of autophagic machinery in response to plasma membrane damage triggered by Candida albicans invasion.
    Pierre Lapaquette, Amandine Ducreux, Etienne Morel, Frédéric Dalle.
      Candida albicans (C. albicans) is an opportunistic pathogen causing infections ranging from superficial to life-threatening dissemination, in which C. albicans is able to translocate through the gut barrier into deeper organs. In its filamentous form (hyphae), C. albicans can invade epithelial cells by two mechanisms: epithelial cell-driven endocytosis and C. albicans-driven active penetration of host cell plasma membrane (PM). Autophagic machinery is known to be involved in the epithelial barrier maintenance, especially the intestinal barrier that is continuously challenged by exposure to the gut microbiota or to xenobiotics. The protective role of autophagy during C. albicans infection has been investigated in myeloid cells, however, far less was known regarding its role during infection of epithelial cells. Here, we demonstrated that key proteins of the autophagic machinery and vesicles presenting features of autophagosomes are recruited at C. albicans invasion sites. These events are associated with host PM damage caused by the active penetration of C. albicans. We showed that ATG5 and ATG16L1 proteins contribute to PM repair mediated by lysosomal membrane exocytosis and participate in protection of epithelial cells' integrity against C. albicans-induced cell death. Our findings extend the knowledge on emerging roles of the autophagic machinery in stress-related membrane dynamics.
    Keywords:  Autophagy; Candida albicans; epithelial cells; lysosomal exocytosis; plasma membrane repair
    DOI:  https://doi.org/10.1080/15548627.2022.2065437
  23. Int J Mol Sci. 2022 Apr 10. pii: 4193. [Epub ahead of print]23(8):
    Autophagy Signaling by Neural-Induced Human Adipose Tissue-Derived Stem Cell-Conditioned Medium during Rotenone-Induced Toxicity in SH-SY5Y Cells.
    Mahesh Ramalingam, Han-Seong Jeong, Jinsu Hwang, Hyong-Ho Cho, Byeong C Kim, Eungpil Kim, Sujeong Jang.
      Rotenone (ROT) inhibits mitochondrial complex I, leading to reactive oxygen species formation, which causes neurodegeneration and alpha-synuclein (α-syn) aggregation and, consequently, Parkinson's disease. We previously found that a neurogenic differentiated human adipose tissue-derived stem cell-conditioned medium (NI-hADSC-CM) was protective against ROT-induced toxicity in SH-SY5Y cells. In the present study, ROT significantly decreased the phospho (p)-mTORC1/total (t)-mTOR, p-mTORC2/t-mTOR, and p-/t-ULK1 ratios and the ATG13 level by increasing the DEPTOR level and p-/t-AMPK ratio. Moreover, ROT increased the p-/t-Akt ratio and glycogen synthase kinase-3β (GSK3β) activity by decreasing the p-/t-ERK1/2 ratios and beclin-1 level. ROT also promoted the lipidation of LC3B-I to LC3B-II by inducing autophagosome formation in Triton X-100-soluble and -insoluble cell lysate fractions. Additionally, the levels of ATG3, 5, 7, and 12 were decreased, along with those of lysosomal LAMP1, LAMP2, and TFEB, leading to lysosomal dysfunction. However, NI-hADSC-CM treatment increased the p-mTORC1, p-mTORC2, p-ULK1, p-Akt, p-ERK1/2, ATG13, and beclin-1 levels and decreased the p-AMPK level and GSK3β activity in response to ROT-induced toxicity. Additionally, NI-hADSC-CM restored the LC3B-I level, increased the p62 level, and normalized the ATG and lysosomal protein amounts to control levels. Autophagy array revealed that the secreted proteins in NI-hADSC-CM could be crucial in the neuroprotection. Taken together, our results showed that the neuroprotective effects of NI-hADSC-CM on the autophagy signaling pathways could alleviate the aggregation of α-syn in Parkinson's disease and other neurodegenerative disorders.
    Keywords:  LC3B; Parkinson’s disease; mTOR; mesenchymal stem cells; rotenone
    DOI:  https://doi.org/10.3390/ijms23084193
  24. Nat Commun. 2022 Apr 19. 13(1): 2025
    Distinct and additive effects of calorie restriction and rapamycin in aging skeletal muscle.
    Daniel J Ham, Anastasiya Börsch, Kathrin Chojnowska, Shuo Lin, Aurel B Leuchtman, Alexander S Ham, Marco Thürkauf, Julien Delezie, Regula Furrer, Dominik Burri, Michael Sinnreich, Christoph Handschin, Lionel A Tintignac, Mihaela Zavolan, Nitish Mittal, Markus A Rüegg.
      Preserving skeletal muscle function is essential to maintain life quality at high age. Calorie restriction (CR) potently extends health and lifespan, but is largely unachievable in humans, making "CR mimetics" of great interest. CR targets nutrient-sensing pathways centering on mTORC1. The mTORC1 inhibitor, rapamycin, is considered a potential CR mimetic and is proven to counteract age-related muscle loss. Therefore, we tested whether rapamycin acts via similar mechanisms as CR to slow muscle aging. Here we show that long-term CR and rapamycin unexpectedly display distinct gene expression profiles in geriatric mouse skeletal muscle, despite both benefiting aging muscles. Furthermore, CR improves muscle integrity in mice with nutrient-insensitive, sustained muscle mTORC1 activity and rapamycin provides additive benefits to CR in naturally aging mouse muscles. We conclude that rapamycin and CR exert distinct, compounding effects in aging skeletal muscle, thus opening the possibility of parallel interventions to counteract muscle aging.
    DOI:  https://doi.org/10.1038/s41467-022-29714-6
  25. Biochim Biophys Acta Mol Basis Dis. 2022 Apr 18. pii: S0925-4439(22)00082-5. [Epub ahead of print] 166412
    The role of autophagy in the metabolism and differentiation of stem cells.
    Maryam Adelipour, Leena Regi Saleth, Saeid Ghavami, Keshav Narayan Alagarsamy, Sanjiv Dhingra, Abdolamir Allameh.
      Autophagy is a very well-coordinated intracellular process that maintains cellular homeostasis under basal conditions by removing unnecessary or dysfunctional components through orderly degradation and recycling. Under pathological conditions, defects in autophagy have been linked to various human disorders, including neurodegenerative disorders and cancer. The role of autophagy in stem cell proliferation, differentiation, self-renewal, and senescence is well documented. Additionally, cancer stem cells (CSCs) play an important role in tumorigenesis, metastasis and tumor relapse and several studies have suggested the involvement of autophagy in the maintenance and invasiveness of CSCs. Hence, considering the modulation of autophagy in normal and cancer stems cells as a therapeutic approach can lead to the development or improvement of regenerative and anti-cancer therapies. Accordingly, modulation of autophagy can be regarded as a target for stem cell-based therapy of diseases with abnormal levels of autophagy. This article is focused on understanding the role of autophagy in stem cell homeostasis with an emphasis on the therapeutic potential of targeting autophagy for future therapies.
    Keywords:  Autophagy; Differentiation; Metabolism; Proliferation; Stem cell
    DOI:  https://doi.org/10.1016/j.bbadis.2022.166412
  26. Autophagy. 2022 Apr 18.
    Autophagy profiling in single cells with open source CellProfiler-based image analysis.
    David S Schüssele, Patricia K Haller, Maximilian L Haas, Catherine Hunter, Katharina Sporbeck, Tassula Proikas-Cezanne.
      Single cell-based analysis of macroautophagy/autophagy is largely achieved through the use of fluorescence microscopy to detect autophagy-related proteins that associate with autophagic membranes and therefore can be quantified as fluorescent puncta. In this context, an automated analysis of the number and size of recognized puncta is preferable to a manual count, because more reliable results can be generated in a short time. Here we present a method for open source CellProfiler software-based analysis for quantitative autophagy assessments using GFP-tagged WIPI1 (WD repeat domain, phosphoinositide interacting 1) images acquired with Airyscan or confocal laser-scanning microscopy. The CellProfiler protocol is provided as a ready-to-use software pipeline, and the creation of this pipeline is detailed in both text and video formats. In addition, we provide CellProfiler pipelines for endogenous SQSTM1/p62 (sequestosome 1) or intracellular lipid droplet (LD) analysis, suitable to assess forms of selective autophagy. All protocols and software pipelines can be quickly and easily adapted for the use of alternative autophagy markers or cell types, and can also be used for high-throughput purposes.
    Keywords:  CellProfiler; SQSTM1/p62; WIPI1; airyscan microscopy; autophagy; lipid droplets
    DOI:  https://doi.org/10.1080/15548627.2022.2065617
  27. Biomolecules. 2022 Apr 09. pii: 559. [Epub ahead of print]12(4):
    Parkin as a Molecular Bridge Linking Alzheimer's and Parkinson's Diseases?
    Frédéric Checler, Cristine Alves da Costa.
      Alzheimer's (AD) and Parkinson's (PD) diseases are two distinct age-related pathologies that are characterized by various common dysfunctions. They are referred to as proteinopathies characterized by ubiquitinated protein accumulation and aggregation. This accumulation is mainly due to altered lysosomal and proteasomal clearing processes and is generally accompanied by ER stress disturbance, autophagic and mitophagic defects, mitochondrial structure and function alterations and enhanced neuronal cell death. Genetic approaches aimed at identifying molecular triggers responsible for familial forms of AD or PD have helped to understand the etiology of their sporadic counterparts. It appears that several proteins thought to contribute to one of these pathologies are also likely to contribute to the other. One such protein is parkin (PK). Here, we will briefly describe anatomical lesions and genetic advances linked to AD and PD as well as the main cellular processes commonly affected in these pathologies. Further, we will focus on current studies suggesting that PK could well participate in AD and thereby act as a molecular bridge between these two pathologies. In particular, we will focus on the transcription factor function of PK and its newly described transcriptional targets that are directly related to AD- and PD-linked cellular defects.
    Keywords:  Alzheimer’s disease; ER stress; PINK1; Parkinson’s disease; XBP1s; autophagy; cell death; mitochondrial dysfunction; mitophagy; p53; parkin
    DOI:  https://doi.org/10.3390/biom12040559
  28. Life Sci. 2022 Apr 19. pii: S0024-3205(22)00252-1. [Epub ahead of print] 120552
    Cancer cell's internal and external warriors: Autophagosomes and exosomes.
    V A Subramanian, Ravindra Bairwa, Pradeep Sharma, Bhawana Bissa.
      "That survival instinct, that will to live, that need to get back to life again, is more powerful than any consideration of taste, decency, politeness, manners, civility, anything. It's such a powerful force." This quote by famous director Danny Boyle is a perfect analogy to describe the cancer cell's inexhaustible drive to persist against all odds. In order to adapt to a hostile environment, the cancer cells rely on multiple mechanisms including immune escape, epithelial to mesenchymal transition, angiogenesis, extravasation, autophagy, exosome release among others. Cancer cells depute their internal and external warriors, autophagosomes and exosomes, to dwell in the belligerent tumor microenvironment. It is quite reasonable for a cancer cell, striving to survive, to invest in pathways that will provide the maximum advantage. Autophagy is an important cellular degradation pathway, while the exosome pathway provides an alternative cargo disposal mechanism to maintain the homeostasis and cell survival. While autophagic degradation provides the essential nutrients to rapidly dividing cells, exosomal secretion ensures that the tumor microenvironment is attuned to accommodate the swiftly expanding tumor mass. Studies have revealed that exosomes secreted by cancer cells can modulate autophagy in recipient cells, while autophagy can influence the biogenesis of exosomes. Autophagy and exosome crosstalk is extremely complex and it is only beginning to be recognized and documented. This review is focused on discussing the roles of autophagy and exosomes in the cancer cell's adaptation to the tumor microenvironment and how the two pathways are coordinately regulated to facilitate cancer cell survival.
    Keywords:  Autophagy; Chemoresistance; Exosomes; Metastasis; Tumor microenvironment
    DOI:  https://doi.org/10.1016/j.lfs.2022.120552
  29. Cells. 2022 Apr 08. pii: 1269. [Epub ahead of print]11(8):
    Steady-State Levels of Miro1 Linked to Phosphorylation at Serine 156 and Mitochondrial Respiration in Dopaminergic Neurons.
    Lisa Schwarz, Julia C Fitzgerald.
      Miro1 has emerged as an interesting target to study Parkinson's disease-relevant pathways since it is a target of PINK1 and Parkin. Miro1 is a mitochondrial GTPase with the primary function of facilitating mitochondrial movement, and its knockout in mice is postnatally lethal. Here, we investigated the effect of the artificial RHOT1/Miro1 S156A mutation since it is a putative PINK1 phosphorylation site shown to be involved in Miro1 degradation and mitochondrial arrest during mitophagy. We gene-edited a homozygous phospho-null Miro1 S156A mutation in induced pluripotent stem cells to study the mutation in human dopaminergic neurons. This mutation causes a significant depletion of Miro1 steady-state protein levels and impairs further Miro1 degradation upon CCCP-induced mitophagy. However, mitochondrial mass measured by Tom20 protein levels, as well as mitochondrial area, are not affected in Miro1 S156A neurons. The mitochondria are slightly lengthened, which is in line with their increased turnover. Under basal conditions, we found no discernable effect of the mutation on mitochondrial movement in neurites. Interestingly, the S156A mutation leads to a significant reduction of mitochondrial oxygen consumption, which is accompanied by a depletion of OXPHOS complexes III and V. These effects are not mirrored by Miro1 knockdown in neuroblastoma cells, but they are observed upon differentiation. Undifferentiated Miro1 S156A neural precursor cells do not have decreased Miro1 levels nor OXPHOS complexes, suggesting that the effect of the mutation is tied to development. In mature dopaminergic neurons, the inhibition of Miro1 Ser156 phosphorylation elicits a mild loss of mitochondrial quality involving reduced mitochondrial membrane potential, which is sufficient to induce compensatory events involving OXPHOS. We suggest that the mechanism governing Miro1 steady-state levels depends on differentiation state and metabolic demand, thus underscoring the importance of this pathway in the pathobiology of Parkinson's disease.
    Keywords:  Miro1; PINK1; Parkinson’s disease; mitochondria
    DOI:  https://doi.org/10.3390/cells11081269
  30. Curr Opin Hematol. 2022 May 01. 29(3): 103-111
    Translational control by heme-regulated elF2α kinase during erythropoiesis.
    Jane-Jane Chen, Shuping Zhang.
      PURPOSE OF REVIEW: HRI is the heme-regulated elF2α kinase that phosphorylates the α-subunit of elF2. Although the role of HRI in inhibiting globin synthesis in erythroid cells is well established, broader roles of HRI in translation have been uncovered recently. This review is to summarize the new discoveries of HRI in stress erythropoiesis and in fetal γ-globin expression.RECENT FINDINGS: HRI and activating transcription factor 4 (ATF4) mRNAs are highly expressed in early erythroblasts. Inhibition of protein synthesis by HRI-phosphorylated elF2α (elF2αP) is necessary to maintain protein homeostasis in both the cytoplasm and mitochondria. In addition, HRI-elF2αP specifically enhances translation of ATF4 mRNA leading to the repression of mechanistic target of rapamycin complex 1 (mTORC1) signaling. ATF4-target genes are most highly activated during iron deficiency to maintain mitochondrial function, redox homeostasis, and to enable erythroid differentiation. HRI is therefore a master translation regulator of erythropoiesis sensing intracellular heme concentrations and oxidative stress for effective erythropoiesis. Intriguingly, HRI-elF2αP-ATF4 signaling also inhibits fetal hemoglobin production in human erythroid cells.
    SUMMARY: The primary function of HRI is to maintain protein homeostasis accompanied by the induction of ATF4 to mitigate stress. Role of HRI-ATF4 in γ-globin expression raises the potential of HRI as a therapeutic target for hemoglobinopathy.
    DOI:  https://doi.org/10.1097/MOH.0000000000000704
  31. FASEB J. 2022 May;36(5): e22317
    Poly(I:C) attenuates myocardial ischemia/reperfusion injury by restoring autophagic function.
    Erya Chen, Haiqing Chang, Rui Gao, Yanhua Qiu, Hai Chen, Xu Cheng, Lu Gan, Shixin Ye-Lehmann, Tao Zhu, Jin Liu, Guo Chen, Chan Chen.
      Polyinosinic-polycytidylic acid (poly(I:C)) is the agonist of Toll-like receptor 3 (TLR3), which participates in innate immune responses under the condition of myocardial ischemia/reperfusion injury (MIRI). It has been shown that poly(I:C) exhibited cardioprotective activities through the PI3K/Akt pathway, which is the main signal transduction pathway during autophagy. However, the precise mechanism by whether poly(I:C) regulates autophagy remains poorly understood. Thus, this study was designed to investigate the therapeutic effect of poly(I:C) against MIRI and the underlying pathway connection with autophagy. We demonstrated that 1.25 and 5 mg/kg poly(I:C) preconditioning significantly reduced myocardial infarct size and cardiac dysfunction. Moreover, poly(I:C) significantly promoted cell survival by restoring autophagy flux and then regulating it to an adequate level Increased autophagy protein Beclin1 and LC3II together with p62 degradation after additional chloroquine. In addition, mRFP-GFP-LC3 adenoviruses exhibited autophagy activity in neonatal rat cardiac myocytes (NRCMs). Mechanistically, poly(I:C) activated the PI3K/AKT/mTOR pathway to induce autophagy, which was abolished by LY294002 (PI3K antagonist), rapamycin (autophagy activator and mTOR inhibitor), or 3-methyladenine (autophagy inhibitor), suggesting either inhibition of the PI3K/Akt/mTOR pathway or autophagy activity interrupt the beneficial effect of poly(I:C) preconditioning. In conclusion, poly(I:C) promotes cardiomyocyte survival from ischemia/reperfusion injury by regulating autophagy via the PI3K/Akt/mTOR pathway.
    Keywords:  Poly(I:C); autophagy; cardio-protection; ischemia/reperfusion injury
    DOI:  https://doi.org/10.1096/fj.202101220RR
  32. Apoptosis. 2022 Apr 18.
    mTOR deletion ameliorates CD4 + T cell apoptosis during sepsis by improving autophagosome-lysosome fusion.
    Hao Wang, Guangxu Bai, Jianwei Chen, Wen Han, Ran Guo, Na Cui.
      Autophagy dysfunction contributes to CD4 + T cell apoptosis during sepsis leading to impairment of adaptive immunity. However, the underlying mechanism is unclear. The mammalian target of rapamycin (mTOR) pathway modulates CD4 + T cell survival during sepsis through mechanisms that are not fully understood. We developed a mouse model of sepsis through cecal ligation and puncture (CLP) to investigate dynamic changes in autophagy in CD4 + T cells. We used T cell specific-mTOR/tuberous sclerosis complex 1 (TSC1)-knockout mice to explore the roles of the mTOR pathway in modulating autophagy during sepsis. We observed reduced fusion of autophagosomes with lysosomes in the CD4 + T cells of CLP mice, which may represent a characteristic feature of autophagy dysfunction. Deletion of mTOR relieved autophagosome-lysosome fusion dysfunction and ameliorated apoptosis of CD4 + T cells in CLP mice, but this rescued phenotype was abolished by treatment with bafilomycin A1, a specific A-L fusion inhibitor. We further explored the underlying molecular mechanism and found that phosphorylation levels of transcription factor EB were significant higher in CLP mice and that expression of A-L fusion protein SNAREs were restricted, both of which were ameliorated by mTOR deletion. Taken together, these results suggest that the mTOR pathway plays a critical role in regulation of CD4 + T-cell apoptosis during sepsis, partly through regulation of A-L fusion-related protein transcription.
    Keywords:  Autophagosome-lysosome fusion; Sepsis; mTOR
    DOI:  https://doi.org/10.1007/s10495-022-01719-y
  33. Front Cell Dev Biol. 2022 ;10 854397
    SLC38A10 Regulate Glutamate Homeostasis and Modulate the AKT/TSC2/mTOR Pathway in Mouse Primary Cortex Cells.
    Rekha Tripathi, Tanya Aggarwal, Frida A Lindberg, Anna H Klemm, Robert Fredriksson.
      Glutamate acts as a critical regulator of neurotransmitter balance, recycling, synaptic function and homeostasis in the brain and glutamate transporters control glutamate levels in the brain. SLC38A10 is a member of the SLC38 family and regulates protein synthesis and cellular stress responses. Here, we uncover the role of SLC38A10 as a transceptor involved in glutamate-sensing signaling pathways that control both the glutamate homeostasis and mTOR-signaling. The culture of primary cortex cells from SLC38A10 knockout mice had increased intracellular glutamate. In addition, under nutrient starvation, KO cells had an impaired response in amino acid-dependent mTORC1 signaling. Combined studies from transcriptomics, protein arrays and metabolomics established that SLC38A10 is involved in mTOR signaling and that SLC38A10 deficient primary cortex cells have increased protein synthesis. Metabolomic data showed decreased cholesterol levels, changed fatty acid synthesis, and altered levels of fumaric acid, citrate, 2-oxoglutarate and succinate in the TCA cycle. These data suggests that SLC38A10 may act as a modulator of glutamate homeostasis, and mTOR-sensing and loss of this transceptor result in lower cholesterol, which could have implications in neurodegenerative diseases.
    Keywords:  SLC38A10; mTOR; primary cortex cultures; solute carriers transporter; starvation
    DOI:  https://doi.org/10.3389/fcell.2022.854397
  34. DNA Cell Biol. 2022 Apr 22.
    Calorie Restriction-Regulated Molecular Pathways and Its Impact on Various Age Groups: An Overview.
    Anuradha Rachakatla, Rajender Rao Kalashikam.
      Calorie restriction (CR) if planned properly with regular exercise at different ages can result in healthy weight loss. CR can also have different beneficial effects on improving lifespan and decreasing the age-associated diseases by regulating physiological, biochemical, and molecular markers. The different pathways regulated by CR include:(1) AMP-activated protein kinase (AMPK), which involves PGC-1α, SIRT1, and SIRT3. AMPK also effects myocyte enhancer factor 2 (MEF2), peroxisome proliferator-activated receptor delta, and peroxisome proliferator-activated receptor alpha, which are involved in mitochondrial biogenesis and lipid oxidation; (2) Forkhead box transcription factor's signaling is related to the DNA repair, lipid metabolism, protection of protein structure, autophagy, and resistance to oxidative stress; (3) Mammalian target of rapamycin (mTOR) signaling, which involves key factors, such as S6 protein kinase-1 (S6K1), mTOR complex-1 (mTORC1), and 4E-binding protein (4E-BP). Under CR conditions, AMPK activation and mTOR inhibition helps in the activation of Ulk1 complex along with the acetyltransferase Mec-17, which is necessary for autophagy; (4) Insulin-like growth factor-1 (IGF-1) pathway downregulation protects against cancer and slows the aging process; (5) Nuclear factor kappa B pathway downregulation decreases the inflammation; and (6) c-Jun N-terminal kinase and p38 kinase regulation as a response to the stress. The acute and chronic CR both shows antidepression and anxiolytic action by effecting ghrelin/GHS-R1a signaling. CR also regulates GSK3β kinase and protects against age-related brain atrophy. CR at young age may show many deleterious effects by effecting different mechanisms. Parental CR before or during conception will also affect the health and development of the offspring by causing many epigenetic modifications that show transgenerational transmission. Maternal CR is associated with intrauterine growth retardation effecting the offspring in their adulthood by developing different metabolic syndromes. The epigenetic changes with response to paternal food supply also linked to offspring health. CR at middle and old age provides a significant preventive impact against the development of age-associated diseases.
    Keywords:  AMPK; ROS; calorie restriction; mTOR
    DOI:  https://doi.org/10.1089/dna.2021.0922
  35. J Cachexia Sarcopenia Muscle. 2022 Apr 17.
    Autophagy in muscle regeneration: potential therapies for myopathies.
    Wei Chen, Yushi Chen, Yuxi Liu, Xinxia Wang.
      Autophagy classically functions as a physiological process to degrade cytoplasmic components, protein aggregates, and/or organelles, as a mechanism for nutrient breakdown, and as a regulator of cellular architecture. Its biological functions include metabolic stress adaptation, stem cell differentiation, immunomodulation and diseases regulation, and so on. Current researches have proved that autophagy dysfunction may contribute to the pathogenesis of some myopathies through impairment of myofibres regeneration. Studies of autophagy inhibition also indicate the importance of autophagy in muscle regeneration, while activation of autophagy can restore muscle function in some myopathies. In this review, we aim to report the mechanisms of action of autophagy on muscle regeneration to provide relevant references for the treatment of regenerating defective myopathies by regulating autophagy. Results have shown that one key mechanism of autophagy regulating the muscle regeneration is to affect the differentiation fate of muscle stem cells (MuSCs), including quiescence maintenance, activation and differentiation. The roles of autophagy (organelle/protein degradation, energy facilitation, and/or other) vary at different myogenic stages of the repair process. When the muscle is in homeostasis, basal autophagy can maintain the quiescence state and stemness of MuSCs by renewing organelle and protein. After injury, the increased autophagy flux contributes to meet biological energy demand of MuSCs during activation and proliferation. By mitochondrial remodelling, autophagy during differentiation can promote the metabolic transformation and balance mitochondrial-mediated apoptosis signals in myoblasts. Autophagy in mature myofibres is also essential for the degradation of necrotic myofibres, and may affect the dynamics of MuSCs by affecting the secretion spectrum of myofibres or the recruitment of supporting cells. Except for myogenic cells, autophagy also plays an important role in regulating the function of non-myogenic cells in the muscle microenvironment, which is also essential for successful muscle recovery. Autophagy can regulate the immune microenvironment during muscle regeneration through the recruitment and polarization of macrophages, while autophagy in endothelial cells can regulate muscle regeneration in an angiogenic or angiogenesis-independent manner. Drug or nutrition targeted autophagy has been preliminarily proved to restore muscle function in myopathies by promoting muscle regeneration, and further understanding the role and mechanism of autophagy in various cell types during muscle regeneration will enable more effective combinatorial therapeutic strategies.
    Keywords:  Autophagy; Microenvironment; MuSCs; Myopathies; Regeneration; Skeletal muscle
    DOI:  https://doi.org/10.1002/jcsm.13000
  36. Front Neurosci. 2022 ;16 853911
    Mitochondria and Other Organelles in Neural Development and Their Potential as Therapeutic Targets in Neurodegenerative Diseases.
    Shuyuan Zhang, Juan Zhao, Zhenzhen Quan, Hui Li, Hong Qing.
      The contribution of organelles to neural development has received increasing attention. Studies have shown that organelles such as mitochondria, endoplasmic reticulum (ER), lysosomes, and endosomes play important roles in neurogenesis. Specifically, metabolic switching, reactive oxygen species production, mitochondrial dynamics, mitophagy, mitochondria-mediated apoptosis, and the interaction between mitochondria and the ER all have roles in neurogenesis. Lysosomes and endosomes can regulate neurite growth and extension. Moreover, metabolic reprogramming represents a novel strategy for generating functional neurons. Accordingly, the exploration and application of mechanisms underlying metabolic reprogramming will be beneficial for neural conversion and regenerative medicine. There is adequate evidence implicating the dysfunction of cellular organelles-especially mitochondria-in neurodegenerative disorders, and that improvement of mitochondrial function may reverse the progression of these diseases through the reinforcement of adult neurogenesis. Therefore, these organelles have potential as therapeutic targets for the treatment of neurodegenerative diseases. In this review, we discuss the function of these organelles, especially mitochondria, in neural development, focusing on their potential as therapeutic targets in neurodegenerative disorders, including Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis.
    Keywords:  antioxidants; metabolic reprogramming; mitochondira; neural development; neurodegenerative disease
    DOI:  https://doi.org/10.3389/fnins.2022.853911
  37. Neurobiol Dis. 2022 Apr 19. pii: S0969-9961(22)00129-2. [Epub ahead of print] 105737
    SOD1 mediates lysosome-to-mitochondria communication and its dysregulation by amyloid-β oligomers.
    Andrés Norambuena, Xuehan Sun, Horst Wallrabe, Ruofan Cao, Naidi Sun, Evelyn Pardo, Nutan Shivange, Dora Bigler Wang, Lisa A Post, Heather A Ferris, Song Hu, Ammasi Periasamy, George S Bloom.
      Altered mitochondrial DNA (mtDNA) occurs in neurodegenerative disorders like Alzheimer's disease (AD); how mtDNA synthesis is linked to neurodegeneration is poorly understood. We previously discovered Nutrient-induced Mitochondrial Activity (NiMA), an inter-organelle signaling pathway where nutrient-stimulated lysosomal mTORC1 activity regulates mtDNA replication in neurons by a mechanism sensitive to amyloid-β oligomers (AβOs), a primary factor in AD pathogenesis (Norambuena et al., 2018). Using 5-ethynyl-2'-deoxyuridine (EdU) incorporation into mtDNA of cultured neurons, along with photoacoustic and mitochondrial metabolic imaging of cultured neurons and mouse brains, we show these effects being mediated by mTORC1-catalyzed T40 phosphorylation of superoxide dismutase 1 (SOD1). Mechanistically, tau, another key factor in AD pathogenesis and other tauopathies, reduced the lysosomal content of the tuberous sclerosis complex (TSC), thereby increasing NiMA and suppressing SOD1 activity and mtDNA synthesis. AβOs inhibited these actions. Dysregulation of mtDNA synthesis was observed in fibroblasts derived from tuberous sclerosis (TS) patients, who lack functional TSC and elevated SOD1 activity was also observed in human AD brain. Together, these findings imply that tau and SOD1 couple nutrient availability to mtDNA replication, linking mitochondrial dysfunction to AD.
    Keywords:  Alzheimer's disease; Amino acids; Insulin; Tau; mTOR
    DOI:  https://doi.org/10.1016/j.nbd.2022.105737
  38. Cells. 2022 Apr 13. pii: 1323. [Epub ahead of print]11(8):
    The Autophagy Process in Cervical Carcinogenesis: Role of Non-Coding-RNAs, Molecular Mechanisms, and Therapeutic Targets.
    Alfredo Lagunas-Martínez, Vicente Madrid-Marina, Claudia Gómez-Cerón, Jessica Deas, Oscar Peralta-Zaragoza.
      Autophagy is a highly conserved multistep lysosomal degradation process in which cellular components are localized to autophagosomes, which subsequently fuse with lysosomes to degrade the sequestered contents. Autophagy serves to maintain cellular homeostasis. There is a close relationship between autophagy and tumor progression, which provides opportunities for the development of anticancer therapeutics that target the autophagy pathway. In this review, we analyze the effects of human papillomavirus (HPV) E5, E6, and E7 oncoproteins on autophagy processes in cervical cancer development. Inhibition of the expression or the activity of E5, E6, and E7 can induce autophagy in cells expressing HPV oncogenes. Thus, E5, E6, and E7 oncoproteins target autophagy during HPV-associated carcinogenesis. Furthermore, noncoding RNA (ncRNA) expression profiling in cervical cancer has allowed the identification of autophagy-related ncRNAs associated with HPV. Autophagy-related genes are essential drivers of autophagy and are regulated by ncRNAs. We review the existing evidence regarding the role of autophagy-related proteins, the function of HPV E5, E6, and E7 oncoproteins, and the effects of noncoding RNA on autophagy regulation in the setting of cervical carcinogenesis. By characterizing the mechanisms behind the dysregulation of these critical factors and their impact on host cell autophagy, we advance understanding of the relationship between autophagy and progression from HPV infection to cervical cancer, and highlight pathways that can be targeted in preventive and therapeutic strategies against cervical cancer.
    Keywords:  E6; E7; HPV E5; autophagy; cervical cancer; ncRNAs; signaling pathways
    DOI:  https://doi.org/10.3390/cells11081323
  39. Cells. 2022 Apr 15. pii: 1359. [Epub ahead of print]11(8):
    Autophagy in Rheumatic Diseases: Role in the Pathogenesis and Therapeutic Approaches.
    Alessandra Ida Celia, Serena Colafrancesco, Cristiana Barbati, Cristiano Alessandri, Fabrizio Conti.
      Autophagy is a lysosomal pathway for the degradation of damaged proteins and intracellular components that promotes cell survival under specific conditions. Apoptosis is, in contrast, a critical programmed cell death mechanism, and the relationship between these two processes influences cell fate. Recent evidence suggests that autophagy and apoptosis are involved in the self-tolerance promotion and in the regulatory mechanisms contributing to disease susceptibility and immune regulation in rheumatic diseases. The aim of this review is to discuss how the balance between autophagy and apoptosis may be dysregulated in multiple rheumatic diseases and to dissect the role of autophagy in the pathogenesis of rheumatoid arthritis, systemic lupus erythematosus, and Sjögren's syndrome. Furthermore, to discuss the potential capacity of currently used disease-modifying antirheumatic drugs (DMARDs) to target and modulate autophagic processes.
    Keywords:  apoptosis; autophagy; rheumatic diseases
    DOI:  https://doi.org/10.3390/cells11081359
  40. Microscopy (Oxf). 2022 Apr 21. pii: dfac020. [Epub ahead of print]
    Detection of Autophagy-Related Structures in Fruiting Bodies of Edible Mushroom, Pleurotus ostreatus.
    Yuma Ozaki, Tadanori Aimi, Norihiro Shimomura.
      Autophagy is involved in various fungal morphogenetic processes. However, there are limited reports regarding the role of autophagy in mushroom fruiting body formation. The purpose of this study was to reveal the autophagy-related structures in mushroom-forming fungi. The edible mushroom Pleurotus ostreatus was used in this study. Transmission electron microscopy revealed double-membrane bounded structures containing cytoplasmic components in the fruiting bodies of this fungus. Some of these double-membrane structures were observed to interact with the vacuoles. Additionally, curved flat cisternae of various lengths were detected in the cytoplasm. The shape, size, and thickness of the limiting membrane of the double-membrane structures and the flat cisternae corresponded well with those of the autophagosomes and the isolation membranes, respectively. Regarding autophagosome formation, a membrane-bound specific zone was detected near the isolation membrane, which appeared to expand along the novel membrane. This is the first detailed report showing autophagy-related structures in P. ostreatus and provides a possible model for autophagosome formation in these filamentous fungi. Mini-abstract Autophagy is involved in fungal morphogenetic processes. The fruiting bodies of edible mushroom Pleurotus ostreatus was observed under a TEM. The present study showed autophagy-related structures in this fungus and provides a possible model for autophagosome formation in filamentous fungi.
    Keywords:   Pleurotus ostreatus ; Autophagosome; Autophagy; Fruiting body formation; Isolation membrane; Ultrastructure
    DOI:  https://doi.org/10.1093/jmicro/dfac020
  41. Front Immunol. 2022 ;13 860262
    Signaling Pathways That Mediate Alveolar Macrophage Activation by Surfactant Protein A and IL-4.
    Belén García-Fojeda, Carlos M Minutti, Carlos Montero-Fernández, Cordula Stamme, Cristina Casals.
      Activation of tissue repair program in macrophages requires the integration of IL-4/IL-13 cytokines and tissue-specific signals. In the lung, surfactant protein A (SP-A) is a tissue factor that amplifies IL-4Rα-dependent alternative activation and proliferation of alveolar macrophages (AMs) through the myosin18A receptor. However, the mechanism by which SP-A and IL-4 synergistically increase activation and proliferation of AMs is unknown. Here we show that SP-A amplifies IL-4-mediated phosphorylation of STAT6 and Akt by binding to myosin18A. Blocking PI3K activity or the myosin18A receptor abrogates SP-A´s amplifying effects on IL-4 signaling. SP-A alone activates Akt, mTORC1, and PKCζ and inactivates GSK3α/β by phosphorylation, but it cannot activate arginase-1 activity or AM proliferation on its own. The combined effects of IL-4 and SP-A on the mTORC1 and GSK3 branches of PI3K-Akt signaling contribute to increased AM proliferation and alternative activation, as revealed by pharmacological inhibition of Akt (inhibitor VIII) and mTORC1 (rapamycin and torin). On the other hand, the IL-4+SP-A-driven PKCζ signaling axis appears to intersect PI3K activation with STAT6 phosphorylation to achieve more efficient alternative activation of AMs. Consistent with IL-4+SP-A-driven activation of mTORC1 and mTORC2, both agonists synergistically increased mitochondrial respiration and glycolysis in AMs, which are necessary for production of energy and metabolic intermediates for proliferation and alternative activation. We conclude that SP-A signaling in AMs activates PI3K-dependent branched pathways that amplify IL-4 actions on cell proliferation and the acquisition of AM effector functions.
    Keywords:  IL-4; PKCζ; Pi3k-akt; mTORC1; macrophage alternative activation; metabolism; proliferation; surfactant protein A
    DOI:  https://doi.org/10.3389/fimmu.2022.860262
  42. Mol Cell. 2022 Apr 21. pii: S1097-2765(22)00253-2. [Epub ahead of print]82(8): 1604-1604.e1
    SnapShot: Organelle degradation.
    Noboru Mizushima.
      Organelles are continuously turned over as part of cellular homeostasis and adaptation. Most organelles, even including the nucleus, are degraded by lysosomes via different pathways, such as macroautophagy, microautophagy, organelle-derived vesicle degradation, and crinophagy. In some specific cases-for example, in lens fiber cells-organelles are degraded by cytosolic phospholipases. To view this SnapShot, open or download the PDF.
    DOI:  https://doi.org/10.1016/j.molcel.2022.03.015
  43. J Agric Food Chem. 2022 Apr 20.
    Downregulation of CASTOR1 Inhibits Heat-Stress-Induced Apoptosis and Promotes Casein and Lipid Synthesis in Mammary Epithelial Cells.
    Zhongchao Gai, Yujiao Wang, Jie Wang, Jiapeng Fu, Lu Tian, Xue Li, Jieqiong Zhao, Guoli Gong.
      Heat stress is one of the most important factors limiting the milk yields of dairy animals. This decline can be attributed to the heat-stress-induced apoptosis of mammary epithelial cells (MECs). The cytosolic arginine sensor for mTORC1 subunit 1 (CASTOR1) is a crucial upstream regulator of the mechanistic target of rapamycin complex 1 (mTORC1) signaling, which has close connections with apoptosis. However, the specific roles of CASTOR1 in regulating the apoptosis and lactation of MECs are still obscure. In the present study, we found that heat stress promotes apoptosis and CASTOR1's expression in HC11 cells. Downregulation of CASTOR1 inhibits heat-stress-induced apoptosis through a ROS-independent pathway. In addition, silencing of CASTOR1 promotes cell proliferation, cell cycle progression, and milk component synthesis, and overexpressing of CASTOR1 reverses these observations. Furthermore, we found that silencing of CASTOR1 contributes to the nuclear transport of SREBP1 and promotes lipid synthesis. This study demonstrates the pivotal roles of CASTOR1 in heat-stress-induced apoptosis and milk component synthesis in MECs.
    Keywords:  CASTOR1; MECs; apoptosis; heat stress; lactation; mTOR
    DOI:  https://doi.org/10.1021/acs.jafc.2c00877
  44. J Fungi (Basel). 2022 Mar 30. pii: 354. [Epub ahead of print]8(4):
    Protein Kinase A Regulates Autophagy-Associated Proteins Impacting Growth and Virulence of Aspergillus fumigatus.
    E Keats Shwab, Praveen R Juvvadi, Shareef K Shaheen, John Allen, Greg Waitt, Erik J Soderblom, Yohannes G Asfaw, M Arthur Moseley, William J Steinbach.
      Cellular recycling via autophagy-associated proteins is a key catabolic pathway critical to invasive fungal pathogen growth and virulence in the nutrient-limited host environment. Protein kinase A (PKA) is vital for the growth and virulence of numerous fungal pathogens. However, the underlying basis for its regulation of pathogenesis remains poorly understood in any species. Our Aspergillus fumigatus PKA-dependent whole proteome and phosphoproteome studies employing advanced mass spectroscopic approaches identified numerous previously undefined PKA-regulated proteins in catabolic pathways. Here, we demonstrate reciprocal inhibition of autophagy and PKA activity, and identify 16 autophagy-associated proteins as likely novel PKA-regulated effectors. We characterize the novel PKA-phosphoregulated sorting nexin Atg20, and demonstrate its importance for growth, cell wall stress response, and virulence of A. fumigatus in a murine infection model. Additionally, we identify physical and functional interaction of Atg20 with previously characterized sorting nexin Atg24. Furthermore, we demonstrate the importance of additional uncharacterized PKA-regulated putative autophagy-associated proteins to hyphal growth. Our data presented here indicate that PKA regulates the autophagy pathway much more extensively than previously known, including targeting of novel effector proteins with fungal-specific functions important for invasive disease.
    Keywords:  Aspergillus; aspergillosis; autophagy; cell wall; filamentous fungi; nutrient sensing; pathogenesis; phosphorylation; protein kinase A; proteomics
    DOI:  https://doi.org/10.3390/jof8040354
  45. Mol Neurobiol. 2022 Apr 19.
    Acetaldehyde Induces Cytotoxicity via Triggering Mitochondrial Dysfunction and Overactive Mitophagy.
    Tingting Yan, Yan Zhao, Zhongyu Jiang, Jiyang Chen.
      Overconsumption of alcohol damages brain tissue and causes cognitive dysfunction. It has been suggested that the neurotoxicity caused by excessive alcohol consumption is largely mediated by acetaldehyde, the most toxic metabolite of ethanol. Evidence shows that acetaldehyde impairs mitochondrial function and induces cytotoxicity of neuronal cells; however, the exact mechanisms are not fully understood. The aim of this study was to investigate the role of mitophagy in acetaldehyde-induced cytotoxicity. It was found that acetaldehyde treatment induced mitophagic responses and caused cytotoxicity in SH-SY5Y cells. The levels of light chain 3 (LC3)-II, Beclin1, autophagy-related protein (Atg) 5 and Atg16L1, PTEN-induced putative kinase (PINK)1, and Parkin were significantly elevated, while the level of p62 was reduced in acetaldehyde-treated cells. Acetaldehyde also promoted the accumulation of PINK1 and Parkin on mitochondria and caused a remarkable decrease of mitochondrial mass. Treatment with autophagy inhibitors prevented the decline of mitochondrial mass and alleviated the cytotoxicity induced by acetaldehyde, suggesting that overactive mitophagy might be an important mechanism contributing to acetaldehyde-induced cytotoxicity. Antioxidant N-acetyl-L-cysteine significantly attenuated the mitophagic responses and alleviated the cytotoxicity induced by acetaldehyde, indicating that oxidative stress was a major mediator of the excessive mitophagy induced by acetaldehyde. Taken together, these findings provided new insights into the role of mitophagy and oxidative stress in acetaldehyde-induced cytotoxicity.
    Keywords:  Acetaldehyde; Drp1; Mitophagy; PINK1; Parkin; Reactive oxygen species
    DOI:  https://doi.org/10.1007/s12035-022-02828-0
  46. FEBS Lett. 2022 Apr 23.
    The Complex Roles of Autophagy in Plant Immunity.
    Nyd Sertsuvalkul, April DeMell, Savithramma P Dinesh-Kumar.
      Plant immunity is the result of multiple distinct cellular processes cooperating with each other to generate immune responses. Autophagy is a conserved cellular recycling process and has well established roles in nutrient starvation responses and cellular homeostasis. Recently, the role of autophagy in immunity has become increasingly evident. However, our knowledge about plant autophagy remains limited and how this fundamental cellular process is involved in plant immunity is still somewhat perplexing. Here, we summarize the current understanding of the positive and negative roles of autophagy in plant immunity and how different microbes exploit this process to their own advantage. The dualistic role of autophagy in plant immunity emphasizes that much remains to be explored in this area.
    Keywords:  Autophagy; ETI; HR-PCD; Immunity; PTI; Pathogen effectors; Reactive oxygen species; Salicylic acid
    DOI:  https://doi.org/10.1002/1873-3468.14356
  47. Biomolecules. 2022 Mar 31. pii: 533. [Epub ahead of print]12(4):
    TORC1 Signaling Controls the Stability and Function of α-Arrestins Aly1 and Aly2.
    Ray W Bowman, Eric M Jordahl, Sydnie Davis, Stefanie Hedayati, Hannah Barsouk, Nejla Ozbaki-Yagan, Annette Chiang, Yang Li, Allyson F O'Donnell.
      Nutrient supply dictates cell signaling changes, which in turn regulate membrane protein trafficking. To better exploit nutrients, cells relocalize membrane transporters via selective protein trafficking. Key in this reshuffling are the α-arrestins, selective protein trafficking adaptors conserved from yeast to man. α-Arrestins bind membrane proteins, controlling the ubiquitination and endocytosis of many transporters. To prevent the spurious removal of membrane proteins, α-arrestin-mediated endocytosis is kept in check through phospho-inhibition. This phospho-regulation is complex, with up to 87 phospho-sites on a single α-arrestin and many kinases/phosphatases targeting α-arrestins. To better define the signaling pathways controlling paralogous α-arrestins, Aly1 and Aly2, we screened the kinase and phosphatase deletion (KinDel) library, which is an array of all non-essential kinase and phosphatase yeast deletion strains, for modifiers of Aly-mediated phenotypes. We identified many Aly regulators, but focused our studies on the TORC1 kinase, a master regulator of nutrient signaling across eukaryotes. We found that TORC1 and its signaling effectors, the Sit4 protein phosphatase and Npr1 kinase, regulate the phosphorylation and stability of Alys. When Sit4 is lost, Alys are hyperphosphorylated and destabilized in an Npr1-dependent manner. These findings add new dimensions to our understanding of TORC1 regulation of α-arrestins and have important ramifications for cellular metabolism.
    Keywords:  Saccharomyces cerevisiae; fluorescence microscopy; kinase; phosphatase; protein degradation; protein trafficking; ubiquitin; vacuole proteases; α-arrestin
    DOI:  https://doi.org/10.3390/biom12040533
  48. Am J Physiol Cell Physiol. 2022 Apr 20.
    UBR5 is a novel regulator of WNK1 stability.
    Ji-Ung Jung, Anwesha Ghosh, Svetlana Earnest, Staci L Deaton, Melanie H Cobb.
      The with no lysine (K) 1 (WNK1) protein kinase maintains cellular ion homeostasis in many tissues through actions on ion cotransporters and channels. Increased accumulation of WNK1 protein leads to pseudohypoaldosteronism type II (PHAII), a form of familial hypertension. WNK1 can be degraded via its adaptor-dependent recruitment to the Cullin3-RBX1 E3 ligase complex by the ubiquitin-proteasome system. Disruption of this process also leads to disease. To determine if this is the primary mechanism of WNK1 turnover, we examined WNK1 protein stability and degradation by measuring its rate of decay after blockade of translation. Here, we show that WNK1 protein degradation exhibits atypical kinetics in Hela cells. Consistent with this apparent complexity, we found that multiple degradative pathways can modulate cellular WNK1 protein amount. WNK1 protein is degraded not only by the proteasome, but also by the lysosome. Non-lysosomal cysteine proteases calpain and caspases also influence WNK1 degradation, as inhibitors of these proteases modestly increased WNK1 protein expression. Importantly, we discovered that the E3 ubiquitin ligase UBR5 interacts with WNK1 and its deficiency results in increased WNK1 protein. Our results further demonstrate that increased WNK1 in UBR5-depleted cells is attributable to reduced lysosomal degradation of WNK1 protein. Taken together, our findings provide insights into the multiplicity of degradative pathways involved in WNK1 turnover and uncover UBR5 as a previously unknown regulator of WNK1 protein stability that leads to lysosomal degradation of WNK1 protein.
    Keywords:  Proteolysis; UBR5; WNK1
    DOI:  https://doi.org/10.1152/ajpcell.00417.2021
  49. Front Oncol. 2022 ;12 857968
    Mitochondrion-Localized SND1 Promotes Mitophagy and Liver Cancer Progression Through PGAM5.
    Shiwei Liang, Chuxu Zhu, Caixia Suo, Haoran Wei, Yingxuan Yu, Xuemei Gu, Liang Chen, Mengqiu Yuan, Shengqi Shen, Shiting Li, Linchong Sun, Ping Gao.
      Staphylococcal nuclease domain-containing protein 1 (SND1) is an evolutionarily conserved multifunctional protein that functions mainly in the nucleus and cytoplasm. However, whether SND1 regulates cellular activity through mitochondrial-related functions remains unclear. Herein, we demonstrate that SND1 is localized to mitochondria to promote phosphoglycerate mutase 5 (PGAM5)-mediated mitophagy. We find that SND1 is present in mitochondria based on mass spectrometry data and verified this phenomenon in different liver cancer cell types by performing organelle subcellular isolation. Specifically, The N-terminal amino acids 1-63 of SND1 serve as a mitochondrial targeting sequence (MTS), and the translocase of outer membrane 70 (TOM 70) promotes the import of SND1 into mitochondria. By immunoprecipitation-mass spectrometry (IP-MS), we find that SND1 interacts with PGAM5 in mitochondria and is crucial for the binding of PGAM5 to dynamin-related protein 1 (DRP1). Importantly, we demonstrate that PGAM5 and SND1-MTS are required for SND1-mediated mitophagy under FCCP and glucose deprivation treatment as well as for SND1-mediated cell proliferation and tumor growth both in vitro and in vivo. Aberrant expression of SND1 and PGAM5 predicts poor outcomes in hepatocellular carcinoma (HCC) patients. Taken together, these findings establish a previously unappreciated role of SND1 and the association of mitochondrion-localized SND1 with PGAM5 in mitophagy and tumor progression.
    Keywords:  PGAM5; SND1; hepatocellular carcinoma; mitophagy; tumor growth
    DOI:  https://doi.org/10.3389/fonc.2022.857968
  50. Geroscience. 2022 Apr 20.
    Cannabidiol induces autophagy and improves neuronal health associated with SIRT1 mediated longevity.
    Zhizhen Wang, Peng Zheng, Xi Chen, Yuanyi Xie, Katrina Weston-Green, Nadia Solowij, Yee Lian Chew, Xu-Feng Huang.
      Autophagy is a catabolic process to eliminate defective cellular molecules via lysosome-mediated degradation. Dysfunctional autophagy is associated with accelerated aging, whereas stimulation of autophagy could have potent anti-aging effects. We report that cannabidiol (CBD), a natural compound from Cannabis sativa, extends lifespan and rescues age-associated physiological declines in C. elegans. CBD promoted autophagic flux in nerve-ring neurons visualized by a tandem-tagged LGG-1 reporter during aging in C. elegans. Similarly, CBD activated autophagic flux in hippocampal and SH-SY5Y neurons. Furthermore, CBD-mediated lifespan extension was dependent on autophagy genes (bec-1, vps-34, and sqst-1) confirmed by RNAi knockdown experiments. C. elegans neurons have previously been shown to accumulate aberrant morphologies, such as beading and blebbing, with increasing age. Interestingly, CBD treatment slowed the development of these features in anterior and posterior touch receptor neurons (TRN) during aging. RNAi knockdown experiments indicated that CBD-mediated age-associated morphological changes in TRNs require bec-1 and sqst-1, not vps-34. Further investigation demonstrated that CBD-induced lifespan extension and increased neuronal health require sir-2.1/SIRT1. These findings collectively indicate the anti-aging benefits of CBD treatment, in both in vitro and in vivo models, and its potential to improve neuronal health and longevity.
    Keywords:  Aging; Autophagy; C. elegans; Cannabidiol; Neurite outgrowth; SIRT1/sir-2.1
    DOI:  https://doi.org/10.1007/s11357-022-00559-7
  51. J Cell Mol Med. 2022 Apr 19.
    FGF21 alleviates pulmonary hypertension by inhibiting mTORC1/EIF4EBP1 pathway via H19.
    Xiuchun Li, Yaxin Zhang, Lihuang Su, Luqiong Cai, Chi Zhang, Jianhao Zhang, Junwei Sun, Mengyu Chai, Mengsi Cai, Qian Wu, Chi Zhang, Xiaoqing Yan, Liangxing Wang, Xiaoying Huang.
      Long non-coding RNAs (lncRNAs) play a significant role in pulmonary hypertension (PH). Our preliminary data showed that hypoxia-induced PH is attenuated by fibroblast growth factor 21 (FGF21) administration. Therefore, we further investigated the regulatory role of long non-coding RNAs in PH treated with FGF21. RNA sequencing analysis and real-time PCR identified a significantly up-regulation of the H19 after FGF21 administration. Moreover, gain- and loss-of-function assays demonstrated that FGF21 suppressed hypoxia-induced proliferation of pulmonary artery smooth muscle cells partially through upregulation of H19. In addition, FGF21 deficiency markedly exacerbated hypoxia-induced increases of pulmonary artery pressure and pulmonary vascular remodelling. In addition, AAV-mediated H19 overexpression reversed the malignant phenotype of FGF21 knockout mice under hypoxia expose. Further investigation uncovered that H19 also acted as an orchestra conductor that inhibited the function of mechanistic target of rapamycin complex 1 (mTORC1) by disrupting the interaction of mTORC1 with eukaryotic translation initiation factor 4E-binding protein 1 (EIF4EBP1). Our work highlights the important role of H19 in PH treated with FGF21 and suggests a mechanism involving mTORC1/EIF4EBP1 inhibition, which may provide a fundamental for clinical application of FGF21 in PH.
    Keywords:  fibroblast growth factor 21; long non-coding RNAs; molecular mechanism; pulmonary hypertension; pulmonary vascular diseases
    DOI:  https://doi.org/10.1111/jcmm.17318
  52. EMBO J. 2022 Apr 22. e111238
    The goodies of chelated fat: iron-regulated lipid droplet biogenesis precedes and preserves mitophagy.
    Marc Liesa.
      The role of iron-regulated mitophagy in lipid metabolism is unclear. Recent work by Long, Sanchez-Martinez et al (2022) shows that iron chelation induces a primary change in lipid metabolism that promotes fat accumulation and precedes mitophagy.
    DOI:  https://doi.org/10.15252/embj.2022111238
  53. Cell Death Dis. 2022 Apr 19. 13(4): 375
    Flubendazole induces mitochondrial dysfunction and DRP1-mediated mitophagy by targeting EVA1A in breast cancer.
    Yongqi Zhen, Zhaoxin Yuan, Jiahui Zhang, Yao Chen, Yuning Fu, Yi Liu, Leilei Fu, Lan Zhang, Xian-Li Zhou.
      Breast cancer is still one of the most common malignancies worldwide and remains a major clinical challenge. We previously reported that the anthelmintic drug flubendazole induced autophagy and apoptosis via upregulation of eva-1 homolog A (EVA1A) in triple-negative breast cancer (TNBC) and was repurposed as a novel anti-tumor agent. However, the detailed underlying mechanisms remain unclear and need further investigation. Here, we found that flubendazole impairs the permeability of the mitochondrial outer membrane and mitochondrial function in breast cancer. Meanwhile, flubendazole increased dynamin-related protein (DRP1) expression, leading to the accumulation of PTEN induced putative kinase 1 (PINK1) and subsequent mitochondrial translocation of Parkin, thereby promoting excessive mitophagy. The resultant excessive mitophagy contributed to mitochondrial damage and dysfunction induced by flubendazole, thus inhibiting breast cancer cells proliferation and migration. Moreover, we demonstrated that excessive DRP1-mediated mitophagy played a critical role in response to the anti-tumor effects of EVA1A in breast cancer. Taken together, our results provide new insights into the molecular mechanisms in relation to the anti-tumor activities of flubendazole, and may be conducive to its rational use in potential clinical applications.
    DOI:  https://doi.org/10.1038/s41419-022-04823-8
  54. Virulence. 2022 Apr 20.
    The autophagy-related proteins FvAtg4 and FvAtg8 are involved in virulence and fumonisin biosynthesis in Fusarium verticillioides.
    Yuejie Wang, Xin Liu, Yujiao Xu, Yiying Gu, Xinyue Zhang, Mengxuan Zhang, Wen Wen, Yin-Won Lee, Jianrong Shi, Sherif Ramzy Mohamed, Amira A Goda, Huijun Wu, Xuewen Gao, Qin Gu.
      Autophagy is the main intracellular degradation system by which cytoplasmic materials are transported to and degraded in the vacuole/lysosome of eukaryotic cells, and it also controls cellular differentiation and virulence in a variety of filamentous fungi. However, the contribution of the autophagic pathway to fungal development and pathogenicity in the important maize pathogen and mycotoxigenic fungus Fusarium verticillioides is still unknown. In this study, we characterized two autophagy-related proteins, FvAtg4 and FvAtg8. The F. verticillioides deletion mutants ΔFvAtg4 and ΔFvAtg8 were impaired in autophagosome formation, aerial hyphal formation, sexual growth, lipid turnover, pigmentation and fungal virulence. Interestingly, ΔFvAtg4 and ΔFvAtg8 were defective in fumonisin B1 (FB1) synthesis, which may have resulted from decreased intracellular levels of alanine in the mutants. Our results indicate that FvAtg4 and FvAtg8 contribute to F. verticillioides pathogenicity by regulating the autophagic pathway to control lipid turnover, fumonisin biosynthesis, and pigmentation during its infectious cycle.
    Keywords:  Autophagy; Fumonisin biosynthesis; Fusarium verticillioides; Virulence
    DOI:  https://doi.org/10.1080/21505594.2022.2066611
  55. J Nutr Biochem. 2022 Apr 18. pii: S0955-2863(22)00087-0. [Epub ahead of print] 109016
    High cholesterol and 27-hydroxycholesterol contribute to phosphorylation of tau protein by impairing autophagy causing learning and memory impairment in C57BL/6J mice.
    Tao Wang, Xiaona Zhang, Yushan Wang, Wen Liu, Lijing Wang, Ling Hao, Mengwei Ju, Rong Xiao.
      Cholesterol and its oxidative derivative 27-hydroxycholesterol (27-OHC), synthesized by CYP27A1, play an important role in Alzheimer's disease (AD) and phosphorylation of tau might be partly responsible for its pathogenesis. To investigate whether cholesterol and 27-OHC affected learning and memory through autophagy-mediated phosphorylation of tau, male C57BL/6J mice were administrated with 2% cholesterol diet, CYP27A1-short-hairpin RNA (CYP27A1-shRNA) and 3-methyladenine (3-MA). The results show that dietary cholesterol induces learning and memory impairment by upregulating the expression of brain CYP27A1 and increasing the levels of 27-OHC and 24S-hydroxycholesterol (24S-OHC). The expressions of total-tau (t-tau), phosphorylated-tau (p-tau) protein, glycogen synthase kinase-3β (GSK-3β) and cyclin-dependent kinase 5 (CDK5) are also significantly upregulated in this group. In addition, reduced expressions of Beclin-1 protein and microtubule-associated protein 1 light chain 3 (LC3B) mRNA, over-expression of mammalian target of rapamycin (mTOR) protein suggest that autophagy is impaired during cholesterol burden. However, using of CYP27A1-shRNA remarkably downregulates the expression of brain CYP27A1. Decreased 27-OHC levels in serum and brain, lower expressions of t-tau and p-tau protein are observed in mice treated with CYP27A1-shRNA+2% cholesterol diet. Furthermore, 3-MA causes lower Beclin-1, higher mTOR and p62 on both gene and protein levels, while the expression of t-tau, p-tau, GSK-3β and CDK5 are upregulated, demonstrating that impaired autophagy disturbs the clearance of tau. These findings suggest that dietary cholesterol induces the accumulation and phosphorylation of tau and the mechanism might be associated with impaired autophagy. And our results indicate 27-OHC might be an importance bridge between cholesterol and cognitive decline.
    Keywords:  Autophagy; CYP27A1; Cholesterol; Learning and memory impairment; Tau phosphorylation
    DOI:  https://doi.org/10.1016/j.jnutbio.2022.109016
  56. Cell Death Dis. 2022 Apr 20. 13(4): 385
    Implication of autophagy in the antifibrogenic effect of Rilpivirine: when more is less.
    Federico Lucantoni, Ana M Benedicto, Aleksandra Gruevska, Ángela B Moragrega, Isabel Fuster-Martínez, Juan V Esplugues, Ana Blas-García, Nadezda Apostolova.
      As the main extracellular matrix-producing cells, activated hepatic stellate cells (HSC) are fundamental mediators of liver fibrosis (LF), and understanding their activation/inactivation mechanisms is paramount to the search for novel therapeutics. The antiretroviral drug Rilpivirine (RPV) has demonstrated a hepatoprotective effect in several animal models of chronic liver injury that is related to its antifibrogenic and apoptotic action in HSC. In the present study, we evaluated whether autophagy is implicated in the hepatoprotective action of RPV, as autophagy plays an important role in HSC transdifferentiation. We employed two standard mouse models of chronic liver injury - fatty liver disease and carbon tetrachloride (CCl4)-induced hepatotoxicity -and cultured HSC activated with the profibrotic cytokine TGF-β. RPV enhanced autophagy in the whole liver of both mouse models and in activated HSC, evident in the protein expression of autophagy markers, increased autophagosome content and lysosomal mass. Moreover, increased autophagic flux was observed in RPV-exposed HSC as revealed by tandem fluorescence-tagged LC3 and p62 and analysis of LC3-II accumulation in cells exposed to the lysosomal inhibitor chloroquine. Importantly, autophagy was involved in the cytotoxic effect of RPV on HSC, though in a differential manner. Pharmacological inhibition of autophagy by 3-methyladenine (3-MA) did not affect the diminishing effect of RPV on viability, while treatment with wortmannin or depletion of specific autophagy proteins (ATG5, Beclin-1 and SQSTM1/p62) rescued the detrimental effect of high concentrations of RPV on the viability of activated HSC. Finally, we also provide evidence that RPV compromises the viability of TGF-β-induced HSC independently of its antifibrogenic effect, observed as reduced collagen 1A1 synthesis, and that this effect does not include RPV´s modulation of autophagy. In summary, as a contributor to the mechanisms involved in the hepatoprotective action of RPV, autophagy may be a good candidate to explore when developing novel therapeutics for LF.
    DOI:  https://doi.org/10.1038/s41419-022-04789-7
  57. Brain Behav Immun. 2022 Apr 13. pii: S0889-1591(22)00109-X. [Epub ahead of print]103 61-62
    Are stressed out goblets cells to blame for functional dyspepsia symptoms?
    H R Wardill, A M Martin.
      
    Keywords:  Autophagy; CRH; Corticotropin-releasing hormone; Functional dyspepsia; Goblet cells; Inflammasome; Mucosal barrier; NLRP6; Stress
    DOI:  https://doi.org/10.1016/j.bbi.2022.04.011
  58. Mol Cell. 2022 Apr 21. pii: S1097-2765(22)00313-6. [Epub ahead of print]82(8): 1390-1397
    Quality Control: Maintaining molecular order and preventing cellular chaos.
    Sonya Neal, Fumiaki Ohtake, Ana Maria Cuervo, Ramanujan S Hegde, Ursula Jakob, Michael Lazarou, Wendy V Gilbert, Zhijian J Chen, Sharon A Tooze, James E Haber, Kylie J Walters, F Ulrich Hartl.
      We asked experts from different fields-from genome maintenance and proteostasis to organelle degradation via ubiquitin and autophagy-"What does quality control mean to you?" Despite their diverse backgrounds, they converge on and discuss the importance of continuous quality control at all levels, context, communication, timing, decisions on whether to repair or remove, and the significance of dysregulated quality control in disease.
    DOI:  https://doi.org/10.1016/j.molcel.2022.04.002
  59. Biochem Pharmacol. 2022 Apr 16. pii: S0006-2952(22)00139-3. [Epub ahead of print] 115045
    Pan-mTOR inhibitors sensitize the senolytic activity of Navitoclax via mTORC2 inhibition-mediated apoptotic signaling.
    Weitong Xu, Tingting Zhao, Honghan Chen, Ning Huang, Hui Gong, Jian Zhang, Yu Yang, Tiepeng Li, Gongchang Zhang, Chuhui Gong, Ming Yang, Hengyi Xiao.
      Compounds with senolysis activity are discovered in recent years, featuring by their capacity to specifically eliminate senescent cells in vitro or in vivo. These compounds, referring to as Senolytics, provide a new method for aging counteraction and probably for geriatric disease amelioration. However, their clinical application is unpractical still, mainly because of the safety issue. In fact, the effective dose range even of the most potent senolytic cannot guarantee the safety requirements application for human being. Here, we report a study which investigated the combinational application of one potential senolytic molecule navitoclax, a Bcl-2 inhibitor with several mTOR inhibitors, to assess the influence of this combination on the senolytic outcome. Our results reveal that pan-mTOR inhibitors can reduce the dosage or timespan of navitoclax necessary for reaching IC50 and LT50 in senescent cells, also extend the lifespan of premature-aged Drosophila and mitigate the aging-related phenotype. Our results also confirmed that mTOR inhibitor sensitized senolytic cell death is apoptotic and pan-mTOR inhibitors PP242 and AZD8055 works more effectively than mTORC1 inhibitor Rapamycin. Mechanically, we verified the crucial role of mTORC2 inhibition contributes sensitization by increasing the expression of the pro-apoptotic protein Bim. In summary, this study firstly exposes the sensitization effect of pan-mTOR inhibitors on navitoclax-induced senolytic apoptosis, therefore providing novel evidence to show the advantage of drug combination on setting senotherapy. It also provides an intriguing clue to demonstrate the value of mTORC2 inhibition for apoptotic death of senescent cells.
    Keywords:  apoptosis; mTORC2; navitoclax; senolytic
    DOI:  https://doi.org/10.1016/j.bcp.2022.115045
  60. Front Cell Dev Biol. 2022 ;10 875124
    USP13: Multiple Functions and Target Inhibition.
    Xiaolong Li, Ge Yang, Wenyao Zhang, Biying Qin, Zifan Ye, Huijing Shi, Xinmeng Zhao, Yihang Chen, Bowei Song, Ziqing Mei, Qi Zhao, Feng Wang.
      As a deubiquitination (DUB) enzyme, ubiquitin-specific protease 13 (USP13) is involved in a myriad of cellular processes, such as mitochondrial energy metabolism, autophagy, DNA damage response, and endoplasmic reticulum-associated degradation (ERAD), by regulating the deubiquitination of diverse key substrate proteins. Thus, dysregulation of USP13 can give rise to the occurrence and development of plenty of diseases, in particular malignant tumors. Given its implications in the stabilization of disease-related proteins and oncology targets, considerable efforts have been committed to the discovery of inhibitors targeting USP13. Here, we summarize an overview of the recent advances of the structure, function of USP13, and its relations to diseases, as well as discovery and development of inhibitors, aiming to provide the theoretical basis for investigation of the molecular mechanism of USP13 action and further development of more potent druggable inhibitors.
    Keywords:  deubiquitination; disease; inhibitor; structure; ubiquitin-specific protease 13
    DOI:  https://doi.org/10.3389/fcell.2022.875124
  61. J Biol Chem. 2022 Apr 18. pii: S0021-9258(22)00385-4. [Epub ahead of print] 101945
    Cellular abundance of sodium phosphate co-transporter SLC20A1/PiT1 and phosphate uptake are controlled post-transcriptionally by ESCRT.
    Christoph Zechner, W Mike Henne, Adwait A Sathe, Chao Xing, Genaro Hernandez, Shengyi Sun, Mi Cheong Cheong.
      Inorganic phosphate is essential for human life. The widely expressed mammalian sodium/phosphate co-transporter SLC20A1/PiT1 mediates phosphate uptake into most cell types; however, while SLC20A1 is required for development, and elevated SLC20A1 expression is associated with vascular calcification and aggressive tumor growth, the mechanisms regulating SLC20A1 protein abundance are unknown. Here we found that SLC20A1 protein expression is low in phosphate-replete cultured cells but is strikingly induced following phosphate starvation, while mRNA expression is high in phosphate-replete cells and only mildly increased by phosphate starvation. To identify regulators of SLC20A1 protein levels, we performed a genome-wide CRISPR-based loss-of-function genetic screen in phosphate-replete cells using SLC20A1 protein induction as readout. Our screen revealed that endosomal sorting complexes required for transport (ESCRT) machinery was essential for proper SLC20A1 protein down-regulation in phosphate-replete cells. We show that SLC20A1 co-localizes with ESCRT, and that ESCRT deficiency increases SLC20A1 protein and phosphate uptake into cells. We also found numerous additional candidate regulators of mammalian phosphate homeostasis, including genes modifying protein ubiquitination and the Krebs cycle and oxidative phosphorylation pathways. Many of these targets have not been previously implicated in this process. We present here a model in which SLC20A1 protein abundance and phosphate uptake are tonically negatively regulated post-transcriptionally in phosphate-replete cells through direct ESCRT-mediated SLC20A1 degradation. Moreover, our screening results provide a comprehensive resource for future studies to elucidate the mechanisms governing cellular phosphate homeostasis. We conclude that genome-wide CRISPR-based genetic screening is a powerful tool to discover proteins and pathways relevant to physiological processes.
    Keywords:  CRISPR/Cas9; Membrane transport; cell metabolism; cell surface protein; endosomal sorting complexes required for transport (ESCRT); genome-wide forward genetic screen; phosphate transporter; protein degradation
    DOI:  https://doi.org/10.1016/j.jbc.2022.101945
  62. Cell Death Dis. 2022 Apr 22. 13(4): 398
    Cisplatin resistance can be curtailed by blunting Bnip3-mediated mitochondrial autophagy.
    Caterina Vianello, Veronica Cocetta, Daniela Catanzaro, Gerald W Dorn, Angelo De Milito, Flavio Rizzolio, Vincenzo Canzonieri, Erika Cecchin, Rossana Roncato, Giuseppe Toffoli, Vincenzo Quagliariello, Annabella Di Mauro, Simona Losito, Nicola Maurea, Scaffa Cono, Gabriele Sales, Luca Scorrano, Marta Giacomello, Monica Montopoli.
      Cisplatin (CDDP) is commonly used to treat a multitude of tumors including sarcomas, ovarian and cervical cancers. Despite recent investigations allowed to improve chemotherapy effectiveness, the molecular mechanisms underlying the development of CDDP resistance remain a major goal in cancer research. Here, we show that mitochondrial morphology and autophagy are altered in different CDDP resistant cancer cell lines. In CDDP resistant osteosarcoma and ovarian carcinoma, mitochondria are fragmented and closely juxtaposed to the endoplasmic reticulum; rates of mitophagy are also increased. Specifically, levels of the mitophagy receptor BNIP3 are higher both in resistant cells and in ovarian cancer patient samples resistant to platinum-based treatments. Genetic BNIP3 silencing or pharmacological inhibition of autophagosome formation re-sensitizes these cells to CDDP. Our study identifies inhibition of BNIP3-driven mitophagy as a potential therapeutic strategy to counteract CDDP resistance in ovarian carcinoma and osteosarcoma.
    DOI:  https://doi.org/10.1038/s41419-022-04741-9
  63. Life Sci Alliance. 2022 Aug;pii: e202101309. [Epub ahead of print]5(8):
    Phospholipids alter activity and stability of mitochondrial membrane-bound ubiquitin ligase MARCH5.
    Lisa Merklinger, Johannes Bauer, Per A Pedersen, Rune Busk Damgaard, J Preben Morth.
      Mitochondrial homeostasis is tightly controlled by ubiquitination. The mitochondrial integral membrane ubiquitin ligase MARCH5 is a crucial regulator of mitochondrial membrane fission, fusion, and disposal through mitophagy. In addition, the lipid composition of mitochondrial membranes can determine mitochondrial dynamics and organelle turnover. However, how lipids influence the ubiquitination processes that control mitochondrial homeostasis remains unknown. Here, we show that lipids common to the mitochondrial membranes interact with MARCH5 and affect its activity and stability depending on the lipid composition in vitro. As the only one of the tested lipids, cardiolipin binding to purified MARCH5 induces a significant decrease in thermal stability, whereas stabilisation increases the strongest in the presence of phosphatidic acid. Furthermore, we observe that the addition of lipids to purified MARCH5 alters the ubiquitination pattern. Specifically, cardiolipin enhances auto-ubiquitination of MARCH5. Our work shows that lipids can directly affect the activity of ubiquitin ligases and suggests that the lipid composition in mitochondrial membranes could control ubiquitination-dependent mechanisms that regulate the dynamics and turnover of mitochondria.
    DOI:  https://doi.org/10.26508/lsa.202101309
  64. Int J Mol Sci. 2022 Apr 07. pii: 4076. [Epub ahead of print]23(8):
    DnaJC7 in Amyotrophic Lateral Sclerosis.
    Allison A Dilliott, Catherine M Andary, Meaghan Stoltz, Andrey A Petropavlovskiy, Sali M K Farhan, Martin L Duennwald.
      Protein misfolding is a common basis of many neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). Misfolded proteins, such as TDP-43, FUS, Matrin3, and SOD1, mislocalize and form the hallmark cytoplasmic and nuclear inclusions in neurons of ALS patients. Cellular protein quality control prevents protein misfolding under normal conditions and, particularly, when cells experience protein folding stress due to the fact of increased levels of reactive oxygen species, genetic mutations, or aging. Molecular chaperones can prevent protein misfolding, refold misfolded proteins, or triage misfolded proteins for degradation by the ubiquitin-proteasome system or autophagy. DnaJC7 is an evolutionarily conserved molecular chaperone that contains both a J-domain for the interaction with Hsp70s and tetratricopeptide domains for interaction with Hsp90, thus joining these two major chaperones' machines. Genetic analyses reveal that pathogenic variants in the gene encoding DnaJC7 cause familial and sporadic ALS. Yet, the underlying ALS-associated molecular pathophysiology and many basic features of DnaJC7 function remain largely unexplored. Here, we review aspects of DnaJC7 expression, interaction, and function to propose a loss-of-function mechanism by which pathogenic variants in DNAJC7 contribute to defects in DnaJC7-mediated chaperoning that might ultimately contribute to neurodegeneration in ALS.
    Keywords:  DNAJC7; Hsp70; Hsp90; J proteins; amyotrophic lateral sclerosis; molecular chaperones; protein misfolding
    DOI:  https://doi.org/10.3390/ijms23084076
  65. Neuron. 2022 Apr 15. pii: S0896-6273(22)00251-3. [Epub ahead of print]
    Programming axonal mitochondrial maintenance and bioenergetics in neurodegeneration and regeneration.
    Xiu-Tang Cheng, Ning Huang, Zu-Hang Sheng.
      Mitochondria generate ATP essential for neuronal growth, function, and regeneration. Due to their polarized structures, neurons face exceptional challenges to deliver mitochondria to and maintain energy homeostasis throughout long axons and terminal branches where energy is in high demand. Chronic mitochondrial dysfunction accompanied by bioenergetic failure is a pathological hallmark of major neurodegenerative diseases. Brain injury triggers acute mitochondrial damage and a local energy crisis that accelerates neuron death. Thus, mitochondrial maintenance defects and axonal energy deficits emerge as central problems in neurodegenerative disorders and brain injury. Recent studies have started to uncover the intrinsic mechanisms that neurons adopt to maintain (or reprogram) axonal mitochondrial density and integrity, and their bioenergetic capacity, upon sensing energy stress. In this review, we discuss recent advances in how neurons maintain a healthy pool of axonal mitochondria, as well as potential therapeutic strategies that target bioenergetic restoration to power neuronal survival, function, and regeneration.
    Keywords:  axonal transport; bioenergetic failure; brain injury; energy deficits; energy metabolism; energy recovery; ischemia; mitochondrial anchoring; mitochondrial quality control; neurodegeneration
    DOI:  https://doi.org/10.1016/j.neuron.2022.03.015
  66. Bioessays. 2022 Apr 20. e2200062
    Tau acetylation reduces its autophagic degradation and is a targetable pathway for human tauopathies (comment on https://doi.org/10.1002/bies.201600224).
    Noah R Johnson.
      
    DOI:  https://doi.org/10.1002/bies.202200062
  67. Pharmaceutics. 2022 Apr 14. pii: 862. [Epub ahead of print]14(4):
    Squalene-Based Nano-Assemblies Improve the Pro-Autophagic Activity of Trehalose.
    Giulia Frapporti, Eleonora Colombo, Hazem Ahmed, Giulia Assoni, Laura Polito, Pietro Randazzo, Daniela Arosio, Pierfausto Seneci, Giovanni Piccoli.
      The disaccharide trehalose is a well-established autophagy inducer, but its therapeutic application is severely hampered by its low potency and poor pharmacokinetic profile. Thus, we targeted the rational design and synthesis of trehalose-based small molecules and nano objects to overcome such issues. Among several rationally designed trehalose-centered putative autophagy inducers, we coupled trehalose via suitable spacers with known self-assembly inducer squalene to yield two nanolipid-trehalose conjugates. Squalene is known for its propensity, once linked to a bioactive compound, to assemble in aqueous media in controlled conditions, internalizing its payload and forming nanoassemblies with better pharmacokinetics. We assembled squalene conjugates to produce the corresponding nanoassemblies, characterized by a hydrodynamic diameter of 188 and 184 nm and a high stability in aqueous media as demonstrated by the measured Z-potential. Moreover, the nanoassemblies were characterized for their toxicity and capability to induce autophagy in vitro.
    Keywords:  autophagy inducers; cancer; nanoassemblies; neurodegeneration; reduction-labile linkers; squalene conjugates; trehalose
    DOI:  https://doi.org/10.3390/pharmaceutics14040862
  68. Front Cell Dev Biol. 2022 ;10 889283
    Editorial: The Regulation of Autophagy Activity in Pathogenic Microorganism Infection.
    Shoulong Deng.
      
    Keywords:  against bacterial; autophagy; immunity; pathogenic microorganism; viral infection
    DOI:  https://doi.org/10.3389/fcell.2022.889283
  69. Pharmaceutics. 2022 Apr 09. pii: 824. [Epub ahead of print]14(4):
    AAV-CRISPR/Cas9 Gene Editing Preserves Long-Term Vision in the P23H Rat Model of Autosomal Dominant Retinitis Pigmentosa.
    Saba Shahin, Hui Xu, Bin Lu, Augustus Mercado, Melissa K Jones, Benjamin Bakondi, Shaomei Wang.
      Retinitis pigmentosa (RP) consists of a group of inherited, retinal degenerative disorders and is characterized by progressive loss of rod photoreceptors and eventual degeneration of cones in advanced stages, resulting in vision loss or blindness. Gene therapy has been effective in treating autosomal recessive RP (arRP). However, limited options are available for patients with autosomal dominant RP (adRP). In vivo gene editing may be a therapeutic option to treat adRP. We previously rescued vision in neonatal adRP rats by the selective ablation of the Rhodopsin S334ter transgene following electroporation of a CRISPR/Cas9 vector. However, the translational feasibility and long-term safety and efficacy of ablation therapy is unclear. To this end, we show that AAV delivery of a CRISPR/Cas9 construct disrupted the Rhodopsin P23H transgene in postnatal rats, which rescued long-term vision and retinal morphology.
    Keywords:  AAV delivery; CRISPR/Cas9; ER-stress; Rhodopsin P23H; allele-specific ablation; autophagy; autosomal dominant retinitis pigmentosa; long-term vision preservation; photoreceptors; rod cells
    DOI:  https://doi.org/10.3390/pharmaceutics14040824
  70. J Interferon Cytokine Res. 2022 Apr;42(4): 170-179
    Vitamin D Attenuates Airway Inflammation in Asthmatic Guinea Pigs Using Mammalian Target of Rapamycin-Mediated Autophagy.
    Lin Zha, Guangji Wu, Hongli Xiao, Yanmin Xiao.
      The purpose of this experiment is to find out the function of Vitamin D (VD) in airway inflammation in asthmatic guinea pigs by regulating mammalian target of rapamycin (mTOR)-mediated autophagy. A total of 40 male guinea pigs were randomly assigned into the Con group, the ovalbumin (OVA)-sensitized group, the VD group, the VD + dimethyl sulfoxide group, and the VD + rapamycin (mTOR inhibitor) group. Then, serum from all groups was harvested for the measurement of immunoglobulin E (IgE), interleukin (IL)-4, and IL-5 levels. Next, bronchoalveolar lavage fluid was collected for cell counting. Moreover, lung tissues were extracted to assess levels of p-mTOR and autophagy factors (LC3B, Beclin1, Atg5, and P62). Compared with the Con group, the OVA group showed elevated levels of IgE, IL-4, and IL-5, increased contents of eosinophils, neutrophil, and lymphocytes, and declined monocytes. And the VD group improved inflammatory reactions in the guinea pigs. Besides, the OVA group showed lower levels of p-mTOR and P62 and higher autophagy levels than the Con group, while the VD group had opposite results. Rapamycin annulled the suppressive role of VD to airway inflammation in asthmatic guinea pigs. VD might inhibit OVA-induced airway inflammation by inducing mTOR activation and downregulating autophagy in asthmatic guinea pigs.
    Keywords:  IgE; airway inflammation; asthma; autophagy; guinea pigs; mTOR; vitamin D
    DOI:  https://doi.org/10.1089/jir.2021.0189
  71. Autophagy. 2022 Apr 18. 1-3
    Apoptosis and autophagy coordinately shape vector tolerance to arbovirus infection.
    Shifan Wang, Huijuan Guo, Feng Ge, Yucheng Sun.
      ABBREVIATIONS: CP: coat protein; MAPK: mitogen-activated protein kinase; PEBP: phosphatidylethanolamine binding protein; TYLCV: tomato yellow leaf curl virus.
    Keywords:  Apoptosis; PEBP; arbovirus; autophagy; immune tolerance
    DOI:  https://doi.org/10.1080/15548627.2022.2062874
  72. Ann Transl Med. 2022 Mar;10(6): 291
    Jian Pi Tiao Gan Yin alleviates obesity phenotypes through mTORC1/SREBP1 signaling in vitro and in vivo.
    Xiaoming Song, Lulu Han, Xiaowan Lin, Minghui Tian, Fenglei Sun, Bo Feng.
      Background: Obesity has been considered as a leading cause of multiple metabolic syndromes, such as type 2 diabetes and hypertension cardiovascular diseases. Jian Pi Tiao Gan Yin (JPTGY), a Chinese herb preparation, is used to treat obesity of liver qi stagnation and spleen deficiency. The mechanism of action of JPTGY in obesity remains unclear. This study evaluated the effect of JPTGY on obesity.Methods: The mechanism of action of JPTGY on obesity was investigated in high-fat diet (HFD)-induced obese mice and palmitic acid-treated 3T3-L1 cells. Lipid droplet accumulation was detected using oil red O staining. Factors associated with lipid accumulation were detected by western blotting.
    Results: Treatment with JPTGY reduced HFD-induced adiposity and body weight gain. JPTGY increased the levels of brown adipose tissue biomarkers in obese mice and palmitic acid-treated 3T3-L1 cells, including peroxisome proliferator-activated receptor gamma coactivator-1-alpha (PGC-1α) and uncoupling protein-1 (UCP-1). Meanwhile, the protein expression of white adipose tissue biomarkers, such as AGT, primary subtalar arthrodesis (PSTA), and endothelin receptor type A (EDNRA), was decreased in obese mice and palmitic acid-treated 3T3-L1 cells. JPTGY affects browning of 3T3-L1 cells through mechanistic target of rapamycin complex 1 (mTORC1) signaling. JPTGY decreased the expression levels of key adipogenic-specific proteins and lipogenic enzymes, including peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein α (C/EBPα), sterol regulatory element binding protein (SREBP), and FAS. Treatment with the mTOR activator MHY reversed JPTGY-mediated protein expression.
    Conclusions: We concluded that JPTGY relieved obesity phenotypes through mTORC1/SREBP1 signaling in vitro and in vivo. JPTGY may benefit the attenuation of obesity.
    Keywords:  Jian Pi Tiao Gan Yin (JPTGY); Obesity; mechanistic target of rapamycin complex 1 (mTORC1); sterol regulatory element-binding protein 1 (SREBP1)
    DOI:  https://doi.org/10.21037/atm-22-685
  73. Cell Death Dis. 2022 Apr 20. 13(4): 381
    Restoration of ER proteostasis attenuates remote apoptotic cell death after spinal cord injury by reducing autophagosome overload.
    Elisa Bisicchia, Roberta Mastrantonio, Annalisa Nobili, Claudia Palazzo, Livia La Barbera, Laura Latini, Francesco Millozzi, Valeria Sasso, Daniela Palacios, Marcello D'Amelio, Maria Teresa Viscomi.
      The pathogenic mechanisms that underlie the progression of remote degeneration after spinal cord injury (SCI) are not fully understood. In this study, we examined the relationship between endoplasmic reticulum (ER) stress and macroautophagy, hereafter autophagy, and its contribution to the secondary damage and outcomes that are associated with remote degeneration after SCI. Using a rat model of spinal cord hemisection at the cervical level, we measured ER stress and autophagy markers in the axotomized neurons of the red nucleus (RN). In SCI animals, mRNA and protein levels of markers of ER stress, such as GRP78, CHOP, and GADD34, increased 1 day after the injury, peaking on Day 5. Notably, in SCI animals, the increase of ER stress markers correlated with a blockade in autophagic flux, as evidenced by the increase in microtubule-associated protein 2 light chain 3 (LC3-II) and p62/SQSTM1 (p62) and the decline in LAMP1 and LAMP2 levels. After injury, treatment with guanabenz protected neurons from UPR failure and increased lysosomes biogenesis, unblocking autophagic flux. These effects correlated with greater activation of TFEB and improved neuronal survival and functional recovery-effects that persisted after suspension of the treatment. Collectively, our results demonstrate that in remote secondary damage, impairments in autophagic flux are intertwined with ER stress, an association that contributes to the apoptotic cell death and functional damage that are observed after SCI.
    DOI:  https://doi.org/10.1038/s41419-022-04830-9
  74. Microb Cell. 2022 Apr 04. 9(4): 72-79
    A hundred spotlights on microbiology: how microorganisms shape our lives.
    Didac Carmona-Gutierrez, Katharina Kainz, Andreas Zimmermann, Sebastian J Hofer, Maria A Bauer, Christoph Ruckenstuhl, Guido Kroemer, Frank Madeo.
      Viral, bacterial, fungal and protozoal biology is of cardinal importance for the evolutionary history of life, ecology, biotechnology and infectious diseases. Various microbiological model systems have fundamentally contributed to the understanding of molecular and cellular processes, including the cell cycle, cell death, mitochondrial biogenesis, vesicular fusion and autophagy, among many others. Microbial interactions within the environment have profound effects on many fields of biology, from ecological diversity to the highly complex and multifaceted impact of the microbiome on human health. Also, biotechnological innovation and corresponding industrial operations strongly depend on microbial engineering. With this wide range of impact in mind, the peer-reviewed and open access journal Microbial Cell was founded in 2014 and celebrates its 100th issue this month. Here, we briefly summarize how the vast diversity of microbiological subjects influences our personal and societal lives and shortly review the milestones achieved by Microbial Cell during the last years.
    Keywords:  biotechnology; infectious diseases; microbiome; microbiota; model organism; open access; symbiosis; yeast
    DOI:  https://doi.org/10.15698/mic2022.04.773
  75. Nutrients. 2022 Apr 12. pii: 1612. [Epub ahead of print]14(8):
    Exploratory Analysis of Selected Components of the mTOR Pathway Reveals Potentially Crucial Associations with Childhood Malnutrition.
    Parag Palit, Md Amran Gazi, Subhasish Das, Md Mehedi Hasan, Zannatun Noor, Jafrin Ferdous, Md Ashraful Alam, Sharika Nuzhat, Md Ridwan Islam, Mustafa Mahfuz, Rashidul Haque, Tahmeed Ahmed.
      Dysregulations in the mammalian target of rapamycin (mTOR) pathway are associated with several human anomalies. We aimed to elucidate possible implications for potential aberrations in the mTOR pathway with childhood malnutrition. We analyzed the activity of phospho-mTORC1 and the expressions of several mTOR pathway genes, namely: MTOR, TSC1, LAMTOR2, RPS6K1 and RICTOR from peripheral blood mononuclear cells isolated from venous blood of children suffering from different forms of malnutrition and compared them with those from healthy children. Significant reduction in the phosphorylation of mTORC1 was noted, as well as a decrease in expression of LAMTOR2 gene and increase in TSC1 gene expression were observed between malnourished children in comparison to the healthy children. The deregulation in the activity of the TSC1 and LAMTOR2 gene was significantly associated with all forms of childhood malnutrition. Our findings provide key insights into possible down-modulation in the overall activity of the mTOR pathway in childhood malnutrition. Further studies focusing on the analysis of a multitude of components involved in the mTOR pathway both at the gene and protein expression levels are required for conclusive evidence for the aforementioned proposition.
    Keywords:  childhood malnutrition; gene expression; mTOR pathway
    DOI:  https://doi.org/10.3390/nu14081612
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