bims-auttor Biomed News
on Autophagy and mTOR
Issue of 2022‒05‒29
58 papers selected by
Viktor Korolchuk, Newcastle University
  1. Role of TFEB in Autophagy and the Pathogenesis of Liver Diseases.
  2. Coordinative regulation of ERAD and selective autophagy in plants.
  3. Vacuolar fragmentation promotes fluxes of microautophagy and micronucleophagy but not of macroautophagy.
  4. O-GlcNAc modification of leucyl-tRNA synthetase 1 integrates leucine and glucose availability to regulate mTORC1 and the metabolic fate of leucine.
  5. Transient visit of STX17 (syntaxin 17) to autophagosomes.
  6. Abemaciclib and Vacuolin-1 induce vacuole-like autolysosome formation - A new tool to study autophagosome-lysosome fusion.
  7. p62 Promotes Survival and Hepatocarcinogenesis in Mice with Liver-Specific NEMO Ablation.
  8. Crosstalk between cilia and autophagy: implication for human diseases.
  9. Cdc14 plans autophagy for meiotic cell divisions.
  10. Autophagy in asthma and chronic obstructive pulmonary disease.
  11. Spatiotemporal feedforward between PKM2 tetramers and mTORC1 prompts mTORC1 activation.
  12. A novel crosstalk between autophagosomes and phagosomes that facilitates the clearance of apoptotic cells.
  13. Theater in the Self-Cleaning Cell: Intrinsically Disordered Proteins or Protein Regions Acting with Membranes in Autophagy.
  14. Hypoxia Induces Autophagy in Human Dendritic Cells: Involvement of Class III PI3K/Vps34.
  15. BAG2 prevents Tau hyperphosphorylation and increases p62/SQSTM1 in cell models of neurodegeneration.
  16. The regulation of NLRP3 inflammasome activation by CCDC50-mediated autophagy.
  17. Hypoxia-Reoxygenation Impairs Autophagy-Lysosomal Machinery in Primary Human Trophoblasts Mimicking Placental Pathology of Early-Onset Preeclampsia.
  18. Japanese Encephalitis Virus NS4A Protein Interacts with PTEN-Induced Kinase 1 (PINK1) and Promotes Mitophagy in Infected Cells.
  19. An intrinsically disordered protein region encoded by the human disease gene CLEC16A regulates mitophagy.
  20. Dyskerin Downregulation Can Induce ER Stress and Promote Autophagy via AKT-mTOR Signaling Deregulation.
  21. Lyso-IP: Uncovering Pathogenic Mechanisms of Lysosomal Dysfunction.
  22. The non-canonical nuclear functions of key players of the PI3K-AKT-MTOR pathway.
  23. F-box protein FBXO41 suppresses breast cancer growth by inducing autophagic cell death through facilitating proteasomal degradation of oncogene SKP2.
  24. Porcine Epidemic Diarrhea Virus Infection Induces Autophagosome Formation but Inhibits Autolysosome Formation during Replication.
  25. The Amyotrophic Lateral Sclerosis M114T PFN1 Mutation Deregulates Alternative Autophagy Pathways and Mitochondrial Homeostasis.
  26. Autophagy protein LC3C binding to phospholipid and interaction with lipid membranes.
  27. Autophagy Modulation in Aggresome Formation: Emerging Implications and Treatments of Alzheimer's Disease.
  28. A bacterial effector counteracts host autophagy by promoting degradation of an autophagy component.
  29. A New Zebrafish Model to Measure Neuronal α-Synuclein Clearance In Vivo.
  30. Regulation of Mitophagy by Sirtuin Family Proteins: A Vital Role in Aging and Age-Related Diseases.
  31. Lysosomal lipid switch sensitises to nutrient deprivation and mTOR targeting in pancreatic cancer.
  32. VMP1 regulates hepatic lipoprotein secretion and NASH independent of autophagy.
  33. RETREG1-mediated ER-phagy activation induced by glucose deprivation alleviates nucleus pulposus cell damage via ER stress pathway.
  34. Nuclear and Cytoplasmatic Players in Mitochondria-Related CNS Disorders: Chromatin Modifications and Subcellular Trafficking.
  35. MoWhi2 Mediates Mitophagy to Regulate Conidiation and Pathogenesis in Magnaporthe oryzae.
  36. Diclofenac: A Nonsteroidal Anti-Inflammatory Drug Inducing Cancer Cell Death by Inhibiting Microtubule Polymerization and Autophagy Flux.
  37. Modulation of Ricin Intoxication by the Autophagy Inhibitor EACC.
  38. Increased glycine contributes to synaptic dysfunction and early mortality in Nprl2 seizure model.
  39. Amino acids control blood glucose levels through mTOR signaling.
  40. Plin2-mediated lipid droplet mobilization accelerates exit from pluripotency by lipidomic remodeling and histone acetylation.
  41. Mechanisms of Mitochondrial Malfunction in Alzheimer's Disease: New Therapeutic Hope.
  42. The ubiquitin ligase Nedd4-2 mediates the regulation of PepT2 by mTORC1 in bovine mammary epithelial cells.
  43. Roles of mTOR in the Regulation of Pancreatic β-Cell Mass and Insulin Secretion.
  44. Association of p53 with Neurodegeneration in Parkinson's Disease.
  45. N, N-dimethylformamide exposure induced liver abnormal mitophagy by targeting miR-92a-1-5p-BNIP3L pathway in vivo and vitro.
  46. A breakdown in microglial metabolic reprogramming causes internalization dysfunction of α-synuclein in a mouse model of Parkinson's disease.
  47. Impact of Raptor and Rictor Deletion on Hippocampal Pathology Following Status Epilepticus.
  48. Infection Dynamics of ATG8 in Leishmania: Balancing Autophagy for Therapeutics.
  49. Safeguarding Lysosomal Homeostasis by DNAJC5/CSPα-Mediated Unconventional Protein Secretion and Endosomal Microautophagy.
  50. Autophagy:a new mechanism for esketamine as a depression therapeutic.
  51. Early bioenergetic and autophagy impairments at the Parkinson's disease synapse.
  52. A meiotic switch in lysosome activity supports spermatocyte development in young flies but collapses with age.
  53. Targeting the lysosome: Mechanisms and treatments for nonalcoholic fatty liver disease.
  54. TRPM2 regulates autophagy to participate in hepatitis B virus replication.
  55. Loss of PTEN-Induced Kinase 1 Regulates Oncogenic Ras-Driven Tumor Growth By Inhibiting Mitochondrial Fission.
  56. An Experimental Study Reveals the Protective Effect of Autophagy against Realgar-Induced Liver Injury via Suppressing ROS-Mediated NLRP3 Inflammasome Pathway.
  57. Beware of Misdelivery: Multifaceted Role of Retromer Transport in Neurodegenerative Diseases.
  58. Myocilin Gene Mutation Induced Autophagy Activation Causes Dysfunction of Trabecular Meshwork Cells.
  1. Biomolecules. 2022 May 06. pii: 672. [Epub ahead of print]12(5):
    Role of TFEB in Autophagy and the Pathogenesis of Liver Diseases.
    Shengmin Yan.
      The transcription factor EB (TFEB) is a master regulator of lysosomal function and autophagy. Mechanistic target of rapamycin (mTOR)-mediated phosphorylation on TFEB is known to regulate TFEB subcellular localization and activity at the lysosomal surface. Recent studies have shown that TFEB also plays a critical role in physiological processes such as lipid metabolism, and dysfunction of TFEB has been observed in the pathogenesis of several diseases. Owing to its ability to improve disease status in murine models, TFEB has attracted attention as a therapeutic target for diseases. In this review, we will present the regulation of TFEB and its role in the pathogenesis of liver diseases, particularly non-alcoholic fatty liver disease (NAFLD).
    Keywords:  TFEB; autophagy; fatty liver disease; metabolism
    DOI:  https://doi.org/10.3390/biom12050672
  2. Essays Biochem. 2022 May 25. pii: EBC20210099. [Epub ahead of print]
    Coordinative regulation of ERAD and selective autophagy in plants.
    Qian Chen, Yaorong Wu, Feifei Yu, Qi Xie.
      Endoplasmic reticulum-associated degradation (ERAD) plays important roles in plant development, hormone signaling, and plant-environment stress interactions by promoting the clearance of certain proteins or soluble misfolded proteins through the ubiquitin-proteasome system. Selective autophagy is involved in the autophagic degradation of protein aggregates mediated by specific selective autophagy receptors. These two major degradation routes co-operate with each other to relieve the cytotoxicity caused by ER stress. In this review, we analyze ERAD and different types of autophagy, including nonselective macroautophagy and ubiquitin-dependent and ubiquitin-independent selective autophagy in plants, and specifically summarize the selective autophagy receptors characterized in plants. In addition to being a part of selective autophagy, ERAD components also serve as their cargos. Moreover, an ubiquitinated substrate can be delivered to two distinguishable degradation systems, while the underlying determinants remain elusive. These excellent findings suggest an interdependent but complicated relationship between ERAD and selective autophagy. According to this point, we propose several key issues that need to be addressed in the future.
    Keywords:  ERAD; plant biology; protein degradation; selective autophagy
    DOI:  https://doi.org/10.1042/EBC20210099
  3. Biochem Biophys Res Commun. 2022 May 11. pii: S0006-291X(22)00717-3. [Epub ahead of print]614 161-168
    Vacuolar fragmentation promotes fluxes of microautophagy and micronucleophagy but not of macroautophagy.
    Tsuneyuki Takuma, Takashi Ushimaru.
      Vacuoles and lysosomes are organelles involved in the degradation of cytoplasmic proteins and organelles. Vacuolar morphology is dynamically regulated by fission and fusion in budding yeast. Vacuolar fusion is elicited in nutrient-depleted conditions and mediated by inactivation of target of rapamycin complex 1 (TORC1) protein kinase. However, it is unknown whether and how vacuolar morphology affects macroautophagy and microautophagy, which are induced by nutrient starvation and TORC1 inactivation. Here, we developed a system to control vacuolar fission in budding yeast. Vacuolar fragmentation promoted microautophagy but not macroautophagy. Vacuolar fragmentation caused multiple nucleus-vacuole junctions. Multiple vacuoles caused by vacuolar fragmentation also improved micronucleophagy (microautophagic degradation of a portion of the nucleus). However, vacuolar morphology did not impact nucleolar remodeling, condensation of the rDNA (rRNA gene) region, or separation of ribosomal DNA from nucleolar proteins, which is evoked by TORC1 inactivation. Thus, this study provides insights into the impacts of vacuolar/lysosomal morphology on macroautophagy and microautophagy.
    Keywords:  Chromosome condensation; Macroautophagy; Microautophagy; Micronucleophagy; Ribosomal DNA (rDNA); Vacuole
    DOI:  https://doi.org/10.1016/j.bbrc.2022.05.021
  4. Nat Commun. 2022 May 25. 13(1): 2904
    O-GlcNAc modification of leucyl-tRNA synthetase 1 integrates leucine and glucose availability to regulate mTORC1 and the metabolic fate of leucine.
    Kibum Kim, Hee Chan Yoo, Byung Gyu Kim, Sulhee Kim, Yulseung Sung, Ina Yoon, Ya Chun Yu, Seung Joon Park, Jong Hyun Kim, Kyungjae Myung, Kwang Yeon Hwang, Sunghoon Kim, Jung Min Han.
      All living organisms have the ability to sense nutrient levels to coordinate cellular metabolism. Despite the importance of nutrient-sensing pathways that detect the levels of amino acids and glucose, how the availability of these two types of nutrients is integrated is unclear. Here, we show that glucose availability regulates the central nutrient effector mTORC1 through intracellular leucine sensor leucyl-tRNA synthetase 1 (LARS1). Glucose starvation results in O-GlcNAcylation of LARS1 on residue S1042. This modification inhibits the interaction of LARS1 with RagD GTPase and reduces the affinity of LARS1 for leucine by promoting phosphorylation of its leucine-binding site by the autophagy-activating kinase ULK1, decreasing mTORC1 activity. The lack of LARS1 O-GlcNAcylation constitutively activates mTORC1, supporting its ability to sense leucine, and deregulates protein synthesis and leucine catabolism under glucose starvation. This work demonstrates that LARS1 integrates leucine and glucose availability to regulate mTORC1 and the metabolic fate of leucine.
    DOI:  https://doi.org/10.1038/s41467-022-30696-8
  5. Autophagy. 2022 May 25. 1-3
    Transient visit of STX17 (syntaxin 17) to autophagosomes.
    Ikuko Koyama-Honda, Noboru Mizushima.
      STX17 (syntaxin 17) mediates autophagosome-lysosome fusion, and the translocation of STX17 to autophagosomes is characteristic of this process. STX17 arrives at autophagosomes when they are closed, stays there for approximately 10 min to promote fusion with lysosomes, and leaves when the autolysosomes are mature. However, the mechanism of this transient visit remains largely unknown. Here, we summarize the current knowledge about this phenomenon, including a recently discovered retrieval mechanism, and discuss remaining questions.Abbreviations: MAM: mitochondria-associated membrane; SNX: sorting nexin; STX17: syntaxin 17.
    Keywords:  ATG9; autophagosome; recycler; sorting nexin; syntaxin 17
    DOI:  https://doi.org/10.1080/15548627.2022.2079337
  6. Biochem Biophys Res Commun. 2022 May 14. pii: S0006-291X(22)00725-2. [Epub ahead of print]614 191-197
    Abemaciclib and Vacuolin-1 induce vacuole-like autolysosome formation - A new tool to study autophagosome-lysosome fusion.
    Yoshinori Tanaka, Hirotsugu Hino, Kosuke Takeya, Masumi Eto.
      Macroautophagy (hereafter autophagy) is a conserved cellular degradation system, impairments in which have been implicated in the development of a wide range of diseases, including cancer and neurodegenerative diseases. Autophagy is mainly comprised of two processes: the formation of autophagosomes and autolysosomes. A detailed understanding of the formation of autophagosomes has been obtained in the past several decades. However, limited information is currently available on the formation of autolysosomes, which may partially be attributed to fewer methods to study the formation of autolysosomes than that of autophagosomes. Abemaciclib (Abe) and vacuolin-1 (Vac) are drugs that suppress the progression of breast cancer and induce characteristic vacuole formation in cells. Since Abe-induced vacuoles have the appearance of autolysosomes, they may be used to examine the formation of autolysosomes. However, it remains unknown whether Abe-/Vac-induced vacuoles are regulated by autophagosome-lysosome fusion. Markers for endosomes, lysosomes, and autophagosomes (Rab7, LAMP1, and mRFP-GFP-LC3, respectively) indicated that Abe-/Vac-induced vacuoles were autolysosomes. Abe and Vac failed to induce vacuolation in ATG16L1-deficient autophagy-null cells. Furthermore, Abe-/Vac-induced vacuolation was suppressed by bafilomycin A1, an inhibitor of autophagosome-lysosome fusion, whereas it was facilitated by rapamycin and the overexpression of Beclin-1, inducers of autophagosome-lysosome fusion. Moreover, vacuole formation was inhibited by the knockdown of progranulin (PGRN), a regulator of autophagosome-lysosome fusion, and promoted by its overexpression. The present results suggest the potential of Abe-/Vac-induced vacuole-like autolysosomes as a tool for evaluating autophagosome-lysosome fusion and examining the effects of PGRN in autophagy.
    Keywords:  Abemaciclib; Autolysosome; Progranulin; Vacuolation; Vacuolin-1
    DOI:  https://doi.org/10.1016/j.bbrc.2022.05.027
  7. Cancers (Basel). 2022 May 15. pii: 2436. [Epub ahead of print]14(10):
    p62 Promotes Survival and Hepatocarcinogenesis in Mice with Liver-Specific NEMO Ablation.
    Vangelis Kondylis, Farina Schneider, Fabian Schorn, Nikos Oikonomou, Beate Katharina Straub, Sabine Werner, Philip Rosenstiel, Manolis Pasparakis.
      SQSTM1/p62 is a multitasking protein that functions as an autophagy receptor, but also as a signaling hub regulating diverse cellular pathways. p62 accumulation in mice with autophagy-deficient hepatocytes mediates liver damage and hepatocarcinogenesis through Nrf2 overactivation, yet the role of the p62-Keap1-Nrf2 axis in cell death and hepatocarcinogenesis in the absence of underlying autophagy defects is less clear. Here, we addressed the role of p62 and Nrf2 activation in a chronic liver disease model, namely mice with liver parenchymal cell-specific knockout of NEMO (NEMOLPC-KO), in which we demonstrate that they show no inherent autophagy impairment. Unexpectedly, systemic p62 ablation aggravated the phenotype and caused early postnatal lethality in NEMOLPC-KO mice. Expression of a p62 mutant (p62ΔEx2-5), which retains the ability to form aggregates and activate Nrf2 signaling, did not cause early lethality, but exacerbated hepatocarcinogenesis in these mice. Our immunohistological and molecular analyses showed that the increased tumor burden was only consistent with increased expression/stability of p62ΔEx2-5 driving Nrf2 hyperactivation, but not with other protumorigenic functions of p62, such as mTOR activation, cMYC upregulation or increased fibrosis. Surprisingly, forced activation of Nrf2 per se did not increase liver injury or tumor burden in NEMOLPC-KO mice, suggesting that autophagy impairment is a necessary prerequisite to unleash the Nrf2 oncogenic potential in mice with autophagy-competent hepatocytes.
    Keywords:  IκB kinase (IKK) complex; Keap1-Nrf2 activation; autophagy; chronic liver disease; genetic mouse models; hepatocarcinogenesis; liver injury
    DOI:  https://doi.org/10.3390/cancers14102436
  8. Autophagy. 2022 May 25. 1-20
    Crosstalk between cilia and autophagy: implication for human diseases.
    Manuela Morleo, Helena L A Vieira, Petra Pennekamp, Alessandro Palma, Liliana Bento-Lopes, Heymut Omran, Susana S Lopes, Duarte C Barral, Brunella Franco.
      Macroautophagy/autophagy is a self-degradative process necessary for cells to maintain their energy balance during development and in response to nutrient deprivation. Autophagic processes are tightly regulated and have been found to be dysfunctional in several pathologies. Increasing experimental evidence points to the existence of an interplay between autophagy and cilia. Cilia are microtubule-based organelles protruding from the cell surface of mammalian cells that perform a variety of motile and sensory functions and, when dysfunctional, result in disorders known as ciliopathies. Indeed, selective autophagic degradation of ciliary proteins has been shown to control ciliogenesis and, conversely, cilia have been reported to control autophagy. Moreover, a growing number of players such as lysosomal and mitochondrial proteins are emerging as actors of the cilia-autophagy interplay. However, some of the published data on the cilia-autophagy axis are contradictory and indicate that we are just starting to understand the underlying molecular mechanisms. In this review, the current knowledge about this axis and challenges are discussed, as well as the implication for ciliopathies and autophagy-associated disorders.
    Keywords:  Autophagy; cilia; human diseases; lysosome; macroautophagy; mitochondria
    DOI:  https://doi.org/10.1080/15548627.2022.2067383
  9. Autophagy. 2022 May 26. 1-2
    Cdc14 plans autophagy for meiotic cell divisions.
    Wenzhi Feng, Orlando Argüello-Miranda, Suhong Qian, Fei Wang.
      The role of meiotic proteasome-mediated degradation has been extensively studied. At the same time, macroautophagy/autophagy only emerged recently as an essential regulator for meiosis progression. Our recent publication showed that autophagy in meiotic cells exhibits a temporal pattern distinct from that in quiescent cells or mitotic cells under prolonged starvation. Importantly, autophagic activity oscillates during meiotic cell divisions, i.e., meiosis I and meiosis II, which can accelerate meiotic progression and increase sporulation efficiency. Our in vitro and in vivo assays revealed that the conserved phosphatase Cdc14 stimulates autophagy initiation during meiotic divisions, specifically in anaphase I and II, when a subpopulation of active Cdc14 relocates to the cytosol and interacts with phagophore assembly sites (PAS) triggering the dephosphorylation of Atg13 to stimulate Atg1 kinase activity and autophagy. Together, our findings reveal a mechanism for the coordination of autophagy activity in the context of meiosis progression.
    Keywords:  Atg1; Atg13; Cdc14; autophagy; meiosis; phosphatase; sporulation
    DOI:  https://doi.org/10.1080/15548627.2022.2080956
  10. Clin Sci (Lond). 2022 May 27. 136(10): 733-746
    Autophagy in asthma and chronic obstructive pulmonary disease.
    Peter J Barnes, Jonathan Baker, Louise E Donnelly.
      Autophagy (or macroautophagy) is a key cellular process that removes damaged molecules (particularly proteins) and subcellular organelles to maintain cellular homeostasis. There is growing evidence that abnormalities in autophagy may contribute to the pathogenesis of many chronic diseases, including asthma and chronic obstructive pulmonary disease (COPD). In asthma, increased autophagy plays a role in promoting type 2 immune responses and eosinophilic inflammation, whereas decreased autophagy may be important in neutrophilic asthma. Acute exposure to cigarette smoke may activate autophagy, resulting in ciliary dysfunction and death of airway epithelial cells, whereas in stable COPD most studies have demonstrated an impairment in autophagy, with reduced autophagic flux and accumulation of abnormal mitochondria (defective mitophagy) and linked to cellular senescence. Autophagy may be increased or decreased in different cell types and depending on the cellular environment, making it difficult to target autophagy therapeutically. Several existing drugs may activate autophagy, including rapamycin, metformin, carbamazepine, cardiac glycosides and statins, whereas others, such as chloroquine, inhibit this process. However, these drugs are nonspecific and more selective drugs are now in development, which may prove useful as novel agents to treat asthma and COPD in the future.
    Keywords:  aggresomes; inflammation; lysosome; macroautophagy; mitophagy; xenophagy
    DOI:  https://doi.org/10.1042/CS20210900
  11. Phys Biol. 2022 May 25.
    Spatiotemporal feedforward between PKM2 tetramers and mTORC1 prompts mTORC1 activation.
    Yu Xia, ShuMing Wang, Chunbo Song, Ruoyu Luo.
      Most mammalian cells couple glucose availability to anabolic processes via the mTORC1 pathway. However, the mechanism by which fluctuations in glucose availability are rapidly translated into mTORC1 signals remains elusive. Here, we show that cells rapidly respond to changes in glucose availability through the spatial coupling of mTORC1 and tetramers of the key glycolytic enzyme pyruvate kinase M2 (PKM2) on lysosomal surfaces in the late G1/S phases. The lysosomal localization of PKM2 tetramers enables rapid increases in local ATP concentrations around lysosomes to activate mTORC1, while bypassing the need to elevate global ATP levels in the entire cell. In essence, this spatial coupling establishes a feedforward loop to enable mTORC1 to rapidly sense and respond to changes in glucose availability. We further demonstrate that this mechanism ensures robust cell proliferation upon fluctuating glucose availability. Thus, we present mechanistic insights into the rapid response of the mTORC1 pathway to changes in glucose availability. The underlying mechanism may be applicable to the control of other cellular processes.
    Keywords:  Cancer metabolism; PKM2; Spatiotemporal Feedforward; mTORC1
    DOI:  https://doi.org/10.1088/1478-3975/ac7372
  12. Autophagy. 2022 May 23.
    A novel crosstalk between autophagosomes and phagosomes that facilitates the clearance of apoptotic cells.
    Zheng Zhou, Omar Peña-Ramos.
      During an animal's life, many cells undergo apoptosis, a form of genetically programmed cell death. These cells are swiftly engulfed by other cells through phagocytosis and subsequently degraded inside phagosomes. Phagocytosis and macroautophagy/autophagy are two different cellular events: whereas phagocytosis is a cell-eat-cell event, autophagy, or "self-eating", occurs within one cell, resulting in the enveloping of protein aggregates or damaged organelles within double-membrane autophagosomes. Despite this critical difference, these two events share common features: (1) both are means of safe garbage disposal; (2) both phagosomes and autophagosomes fuse to lysosomes, which drive the degradation of their contents; and (3) both events facilitate the recycling of biological materials. Previously, whether autophagosomes per se directly participate in the degradation of apoptotic cells was unknown, although autophagy proteins were implicated in apoptotic cell clearance. We recently discovered that autophagosomes fuse with phagosomes and contribute to the degradation of apoptotic cells.
    Keywords:  Autophagosomes, C. elegans; CED-1; LGG-1/LGG-2; crosstalk; degradation of apoptotic cells; membrane fusion; membrane signaling; phagocytosis; phagosomes
    DOI:  https://doi.org/10.1080/15548627.2022.2080384
  13. Membranes (Basel). 2022 Apr 24. pii: 457. [Epub ahead of print]12(5):
    Theater in the Self-Cleaning Cell: Intrinsically Disordered Proteins or Protein Regions Acting with Membranes in Autophagy.
    Hana Popelka, Vladimir N Uversky.
      Intrinsically disordered proteins and protein regions (IDPs/IDPRs) are mainly involved in signaling pathways, where fast regulation, temporal interactions, promiscuous interactions, and assemblies of structurally diverse components including membranes are essential. The autophagy pathway builds, de novo, a membrane organelle, the autophagosome, using carefully orchestrated interactions between proteins and lipid bilayers. Here, we discuss molecular mechanisms related to the protein disorder-based interactions of the autophagy machinery with membranes. We describe not only membrane binding phenomenon, but also examples of membrane remodeling processes including membrane tethering, bending, curvature sensing, and/or fragmentation of membrane organelles such as the endoplasmic reticulum, which is an important membrane source as well as cargo for autophagy. Summary of the current state of knowledge presented here will hopefully inspire new studies. A profound understanding of the autophagic protein-membrane interface is essential for advancements in therapeutic interventions against major human diseases, in which autophagy is involved including neurodegeneration, cancer as well as cardiovascular, metabolic, infectious, musculoskeletal, and other disorders.
    Keywords:  amphipathic helix; disorder prediction; hydrophobic finger; intrinsically disordered protein; intrinsically disordered protein region; lipid bilayer; membrane binding; membrane fragmentation; membrane remodeling; membrane tethering; protein–protein interactions
    DOI:  https://doi.org/10.3390/membranes12050457
  14. Cells. 2022 May 19. pii: 1695. [Epub ahead of print]11(10):
    Hypoxia Induces Autophagy in Human Dendritic Cells: Involvement of Class III PI3K/Vps34.
    Sara Monaci, Federica Coppola, Daniela Rossi, Gaia Giuntini, Irene Filippi, Giuseppe Marotta, Silvano Sozzani, Fabio Carraro, Antonella Naldini.
      Hypoxia is a component of both physiological and pathological conditions, including inflammation, solid tumors, and lymphoid tissues, where O2 demand is not balanced by O2 supply. During their lifespan, dendritic cells (DCs) are exposed to different pO2 and activate different adaptive responses, including autophagy, to preserve their viability and functions. Autophagy plays multiple roles in DC physiology. Very recently, we demonstrated that hypoxia shapes autophagy in DCs upon their differentiation state. Here, we proposed a role for PI3Ks, and especially class III PI3K/Vps34, that could be relevant in hypoxia-induced autophagy, in either immature or mature DCs. Hypoxia inhibited mTOR phosphorylation and activated a pro-autophagic program. By using different pharmacological inhibitors, we demonstrated that hypoxia-induced autophagy was mediated by PI3Ks, especially by Vps34. Furthermore, Vps34 expression was enhanced by LPS, a TLR4 ligand, along with the promotion of autophagy under hypoxia. The Vps34 inhibitor, SAR405, abolished hypoxia-induced autophagy, inhibited pro-survival signaling and viability, and increased the expression of proinflammatory cytokines. Our results underlined the impact of autophagy in the maintenance of DC homeostasis at both cell survival and inflammatory response levels, therefore, contributing to a better understanding of the significance of autophagy in DC physiology and pathology.
    Keywords:  PI3K; SAR405; autophagy; dendritic cell; hypoxia
    DOI:  https://doi.org/10.3390/cells11101695
  15. Mol Biol Rep. 2022 May 25.
    BAG2 prevents Tau hyperphosphorylation and increases p62/SQSTM1 in cell models of neurodegeneration.
    Raquel S Lima, Daniel C Carrettiero, Merari F R Ferrari.
      BACKGROUND: Protein aggregates are pathological hallmarks of many neurodegenerative diseases, however the physiopathological role of these aggregates is not fully understood. Protein quality control has a pivotal role for protein homeostasis and depends on specific chaperones. The co-chaperone BAG2 can target phosphorylated Tau for degradation by an ubiquitin-independent pathway, although its possible role in autophagy was not yet elucidated. In view of this, the aim of the present study was to investigate the association among protein aggregation, autophagy and BAG2 levels in cultured cells from hippocampus and locus coeruleus as well as in SH-SY5Y cell line upon different protein aggregation scenarios induced by rotenone, which is a flavonoid used as pesticide and triggers neurodegeneration.METHODS AND RESULTS: The present study showed that rotenone exposure at 0.3 nM for 48 h impaired autophagy prior to Tau phosphorylation at Ser199/202 in hippocampus but not in locus coeruleus cells, suggesting that distinct neuron cells respond differently to rotenone toxicity. Rotenone induced Tau phosphorylation at Ser199/202, together with a decrease in the endogenous BAG2 protein levels in SH-SY5Y and hippocampus cell culture, which indicates that rotenone and Tau hyperphosphorylation can affect this co-chaperone. Finally, it has been shown that BAG2 overexpression, increased p62/SQSTM1 levels in cells from hippocampus and locus coeruleus, stimulated LC3II recycling as well as prevented the raise of phosphorylated Tau at Ser199/202 in hippocampus.
    CONCLUSIONS: Results demonstrate a possible role for BAG2 in degradation pathways of specific substrates and its importance for the study of cellular aspects of neurodegenerative diseases.
    Keywords:  Alpha-synuclein; Alzheimer’s disease; Autophagy; Hyperphosphorylated Tau; Parkinson’s disease; Rotenone
    DOI:  https://doi.org/10.1007/s11033-022-07577-w
  16. Autophagy. 2022 May 27. 1-2
    The regulation of NLRP3 inflammasome activation by CCDC50-mediated autophagy.
    Panpan Hou, Tian Tian, Penghui Jia, Yuxin Lin, Zibo Li, Yicheng Wang, Yu Ye, Chunmei Li, Deyin Guo.
      The assembly of the NLRP3 inflammasome can be initiated by a wide range of stimuli including exogenous infection as well as endogenous damage. Therefore, the tight regulation of the NLRP3 inflammasome is crucial for the host to resist microbial invasion and maintain homeostasis. Our recent work has identified a negative regulator of NLRP3-mediated inflammation, namely CCDC50 (coiled-coil domain containing protein 50). CCDC50 can be induced by NLRP3 agonists and then functions as a macroautophagy/autophagy cargo receptor to recognize K63-polyubiquitinated NLRP3 and deliver it to MAP1LC3/LC3-conjugated phagophores for degradation. CCDC50 inhibits the polymerization of NLRP3 and the recruitment of PYCARD/ASC, consequently suppressing the assembly of inflammasomes. ccdc50-knockout mice are more susceptible to dextran-sulfate (DSS)-induced colitis and exhibit more severe gut inflammation with elevated NLRP3 inflammasome activity, suggesting a protective role of CCDC50 in the pathology and progression of inflammatory bowel disease (IBD). Our finding reveals a function of autophagy-related proteins in the regulation of NLRP3-mediated inflammation, thus demonstrating the intricate crosstalk between autophagy and inflammation.
    Keywords:  Autophagy receptor; CCDC50; NLRP3 inflammasome; immune homeostasis; inflammatory bowel disease
    DOI:  https://doi.org/10.1080/15548627.2022.2080957
  17. Int J Mol Sci. 2022 May 18. pii: 5644. [Epub ahead of print]23(10):
    Hypoxia-Reoxygenation Impairs Autophagy-Lysosomal Machinery in Primary Human Trophoblasts Mimicking Placental Pathology of Early-Onset Preeclampsia.
    Shibin Cheng, Zheping Huang, Sukanta Jash, Kathleen Wu, Shigeru Saito, Akitoshi Nakashima, Surendra Sharma.
      We have previously described that placental activation of autophagy is a central feature of normal pregnancy, whereas autophagy is impaired in preeclampsia (PE). Here, we show that hypoxia-reoxygenation (H/R) treatment dysregulates key molecules that maintain autophagy-lysosomal flux in primary human trophoblasts (PHTs). Ultrastructural analysis using transmission electron microscopy reveals a significant reduction in autophagosomes and autolysosomes in H/R-exposed PHTs. H/R-induced accumulation of protein aggregates follows a similar pattern that occurs in PHTs treated with a lysosomal disruptor, chloroquine. Importantly, the placenta from early-onset PE deliveries exhibits the same features as seen in H/R-treated PHTs. Taken together, our results indicate that H/R disrupts autophagic machinery in PHTs and that impaired autophagy in the placenta from early-onset PE deliveries mimics the events in H/R-treated PHTs. Notably, assessment of key regulators at each stage of autophagic processes, especially lysosomal integrity, and verification of autophagic ultrastructure are essential for an accurate evaluation of autophagy activity in human trophoblasts and placental tissue from PE deliveries.
    Keywords:  autolysosome; autophagosome; autophagy-related proteins; electron microscopy; preeclampsia; proteasome; trophoblasts
    DOI:  https://doi.org/10.3390/ijms23105644
  18. Microbiol Spectr. 2022 May 23. e0083022
    Japanese Encephalitis Virus NS4A Protein Interacts with PTEN-Induced Kinase 1 (PINK1) and Promotes Mitophagy in Infected Cells.
    Anshu Agarwal, Mohd Faraz Alam, Brohmomoy Basu, Sabyasachi Pattanayak, Shailendra Asthana, Gulam Hussain Syed, Manjula Kalia, Sudhanshu Vrati.
      The nonstructural protein 4A (NS4A) of flaviviruses has been implicated as a "central organizer" of the membrane-bound replication complex during virus replication. However, its role in the host responses to virus infection is not understood. Using the yeast-two-hybrid library screen, we identified a multitude of host proteins interacting with the Japanese encephalitis virus (JEV) NS4A protein. Several of these interacting proteins are known to localize to the mitochondria. One of these proteins was PTEN-induced kinase 1 (PINK1), a serine/threonine-protein kinase known for its role in mitophagy. Here, we demonstrate the JEV-NS4A localization to the mitochondria and its interaction with PINK1 in Huh7 cells during JEV infection. The JEV-infected cells showed an enhanced mitophagy flux with a concomitant decline in the mitochondrial mass. We present data showing that JEV-NS4A alone was sufficient to induce mitophagy. Interference with mitochondrial fragmentation and mitophagy resulted in reduced virus propagation. Overall, our study provides the first evidence of mitochondrial quality control dysregulation during JEV infection, largely mediated by its NS4A protein. IMPORTANCE The JEV-infected mammalian cells show an enhanced mitophagy flux with a concomitant decline in the mitochondrial mass. We show that the NS4A protein of JEV localized to the mitochondria and interacted with PINK1 in Huh7 cells during infection with the virus and demonstrate that JEV-NS4A alone is sufficient to induce mitophagy. The study provides the first evidence of mitochondrial quality control dysregulation during JEV infection, largely mediated by its NS4A protein.
    Keywords:  flavivirus; mitochondria; protein interactome; yeast-two-hybrid
    DOI:  https://doi.org/10.1128/spectrum.00830-22
  19. Autophagy. 2022 May 23.
    An intrinsically disordered protein region encoded by the human disease gene CLEC16A regulates mitophagy.
    Morgan A Gingerich, Xueying Liu, Biaoxin Chai, Gemma L Pearson, Michael P Vincent, Tracy Stromer, Jie Zhu, Vaibhav Sidarala, Aaron Renberg, Debashish Sahu, Daniel J Klionsky, Santiago Schnell, Scott A Soleimanpour.
      CLEC16A regulates mitochondrial health through mitophagy and is associated with over 20 human diseases. However, the key structural and functional regions of CLEC16A, and their relevance for human disease, remain unknown. Here, we report that a disease-associated CLEC16A variant lacks a C-terminal intrinsically disordered protein region (IDPR) that is critical for mitochondrial quality control. IDPRs comprise nearly half of the human proteome, yet their mechanistic roles in human disease are poorly understood. Using carbon detect NMR, we find that the CLEC16A C terminus lacks secondary structure, validating the presence of an IDPR. Loss of the CLEC16A C-terminal IDPR in vivo impairs mitophagy, mitochondrial function, and glucose-stimulated insulin secretion, ultimately causing glucose intolerance. Deletion of the CLEC16A C-terminal IDPR increases CLEC16A ubiquitination and degradation, thus impairing assembly of the mitophagy regulatory machinery. Importantly, CLEC16A stability is dependent on proline bias within the C-terminal IDPR, but not amino acid sequence order or charge. Together, we elucidate how an IDPR in CLEC16A regulates mitophagy and implicate pathogenic human gene variants that disrupt IDPRs as novel contributors to diabetes and other CLEC16A-associated diseases.
    Keywords:  Diabetes; NMR; insulin; mitophagy; splicing
    DOI:  https://doi.org/10.1080/15548627.2022.2080383
  20. Biomedicines. 2022 May 08. pii: 1092. [Epub ahead of print]10(5):
    Dyskerin Downregulation Can Induce ER Stress and Promote Autophagy via AKT-mTOR Signaling Deregulation.
    Daniela Maiello, Marianna Varone, Rosario Vicidomini, Valentina Belli, Marina De Rosa, Paola Dama, Maria Furia, Mimmo Turano.
      Dyskerin is an evolutionarily conserved nucleolar protein implicated in a wide range of fundamental biological roles, including telomere maintenance and ribosome biogenesis. Germline mutations of DKC1, the human gene encoding dyskerin, cause the hereditary disorders known as X-linked dyskeratosis congenita (X-DC). Moreover, dyskerin is upregulated in several cancers. Due to the pleiotropic functions of dyskerin, the X-DC clinical features overlap with those of both telomeropathies and ribosomopathies. In this paper, we evaluate the telomerase-independent effects of dyskerin depletion on cellular physiology by using inducible DCK1 knockdown. This system allows the downregulation of DKC1 expression within a short timeframe. We report that, in these cellular systems, dyskerin depletion induces the accumulation of unfolded/misfolded proteins in the endoplasmic reticulum, which in turn induces the activation of the PERK branch of the unfolded protein response. We also demonstrate that the PERK-eIF2a-ATF4-CHOP signaling pathway, activated by dyskerin downregulation, triggers a functional autophagic flux through the inhibition of the PI3K/AKT/mTOR pathway. By revealing a novel unpredicted connection between the loss of dyskerin, autophagy and UPR, our results establish a firm link between the lowering of dyskerin levels and the activation of the ER stress response, that plays a key role in the pathogenesis of several diseases.
    Keywords:  DKC1; LC3 puncta; UPR; autophagic flux; dyskeratosis congenita
    DOI:  https://doi.org/10.3390/biomedicines10051092
  21. Biomolecules. 2022 Apr 21. pii: 616. [Epub ahead of print]12(5):
    Lyso-IP: Uncovering Pathogenic Mechanisms of Lysosomal Dysfunction.
    Chase Chen, Ellen Sidransky, Yu Chen.
      Lysosomes are ubiquitous membrane-bound organelles found in all eukaryotic cells. Outside of their well-known degradative function, lysosomes are integral in maintaining cellular homeostasis. Growing evidence has shown that lysosomal dysfunction plays an important role not only in the rare group of lysosomal storage diseases but also in a host of others, including common neurodegenerative disorders, such as Alzheimer disease and Parkinson disease. New technological advances have significantly increased our ability to rapidly isolate lysosomes from cells in recent years. The development of the Lyso-IP approach and similar methods now allow for lysosomal purification within ten minutes. Multiple studies using the Lyso-IP approach have revealed novel insights into the pathogenic mechanisms of lysosomal disorders, including Niemann-Pick type C disease, showing the immense potential for this technique. Future applications of rapid lysosomal isolation techniques are likely to greatly enhance our understanding of lysosomal dysfunction in rare and common neurodegeneration causes.
    Keywords:  Niemann-Pick type C; lysosome isolation; lysosomes
    DOI:  https://doi.org/10.3390/biom12050616
  22. J Cell Physiol. 2022 May 26.
    The non-canonical nuclear functions of key players of the PI3K-AKT-MTOR pathway.
    Sakshi Gupta, Mukund Kumar, Soumi Chaudhuri, Arun Kumar.
      The PI3K-AKT-MTOR signal transduction pathway is one of the essential signalling cascades within the cell due to its involvement in many vital functions. The pathway initiates with the recruitment of phosphatidylinositol-3 kinases (PI3Ks) onto the plasma membrane, generating phosphatidylinositol-3,4,5-triphosphate [PtdIns(3,4,5)P3 ] and subsequently activating AKT. Being the central node of the PI3K network, AKT activates the mechanistic target of rapamycin kinase complex 1 (MTORC1) via Tuberous sclerosis complex 2 inhibition in the cytoplasm. Although the cytoplasmic role of the pathway has been widely explored for decades, we now know that most of the effector molecules of the PI3K axis diverge from the canonical route and translocate to other cell organelles including the nucleus. The presence of phosphoinositides (PtdIns) inside the nucleus itself indicates the existence of a nuclear PI3K signalling. The nuclear localization of these signaling components is evident in regulating many nuclear processes like DNA replication, transcription, DNA repair, maintenance of genomic integrity, chromatin architecture, and cell cycle control. Here, our review intends to present a comprehensive overview of the nuclear functions of the PI3K-AKT-MTOR signaling biomolecules.
    Keywords:  MTORC2; PDK1; PTEN; RHEB; TSC1; inhibitors; nuclear entry; therapeutics
    DOI:  https://doi.org/10.1002/jcp.30782
  23. Int J Biochem Cell Biol. 2022 May 19. pii: S1357-2725(22)00073-5. [Epub ahead of print] 106228
    F-box protein FBXO41 suppresses breast cancer growth by inducing autophagic cell death through facilitating proteasomal degradation of oncogene SKP2.
    Yashika Agrawal, Tanisha Sharma, Sehbanul Islam, Kaustubh S Nadkarni, Manas Kumar Santra.
      F-box proteins form SCF (Cullin1, SKP1 and F-box-protein) ubiquitin ligase complexes to ubiquitinate cellular proteins. They play key role in several biological processes, including cell cycle progression, cellular signaling, stress response and cell death pathways. Therefore, deregulation of F-box proteins is closely associated with cancer progression. However, the role of most of the F-box proteins, including FBXO41, in cancer progression remains elusive. Here, we unravel the role of FBXO41 in cancer progression. We show that FBXO41 suppresses cancer cell proliferation and tumor growth by inducing autophagic cell death through an alternative pathway. Results revealed that FBXO41-mediated autophagic cell death induction is dependent on accumulation of cell cycle checkpoint protein p21. We found that FBXO41 increases the expression levels of p21 at the post-translational level by promoting the proteasomal degradation of SKP2, an oncogenic F-box protein. Mechanistically, FBXO41 along with p21 disrupts the inhibitory BCL2 (anti-apoptotic protein)-Beclin1 (autophagy initiating factor) complex of autophagy induction to release Beclin1, thereby inducing autophagy. Overall, the present study establishes a new FBXO41-SKP2-p21 axis for induction of autophagic cell death to prevent cancer growth, which could be explored to develop promising cancer therapeutics.
    Keywords:  Apoptosis independent autophagy; Autophagic cell death; F-box proteins; SKP2; Stress independent autophagy; Tumor suppressor
    DOI:  https://doi.org/10.1016/j.biocel.2022.106228
  24. Viruses. 2022 May 15. pii: 1050. [Epub ahead of print]14(5):
    Porcine Epidemic Diarrhea Virus Infection Induces Autophagosome Formation but Inhibits Autolysosome Formation during Replication.
    Jae-Yeon Park, Jihoon Ryu, Eui-Ju Hong, Hyun-Jin Shin.
      In this study, we investigated the correlation between the mechanism involved in porcine epidemic diarrhea virus (PEDV) replication and autophagic flux. In this study, we found that as PEDV replicated, production of LC3-II was significantly induced up to 24 h post-infection (hpi). Interestingly, although there was significant production of LC3-II, greater p62 accumulation was simultaneously found. Pretreatment with rapamycin significantly induced PEDV replication, but autolysosome formation was reduced. These results were confirmed by the evaluation of ATG5/ATG12 and LAMP1/LAMP2. Taken together, we conclude that PEDV infection induces autophagosome formation but inhibits autolysosome formation during replication.
    Keywords:  PEDV replication; autophagy; autophagy flux
    DOI:  https://doi.org/10.3390/v14051050
  25. Int J Mol Sci. 2022 May 19. pii: 5694. [Epub ahead of print]23(10):
    The Amyotrophic Lateral Sclerosis M114T PFN1 Mutation Deregulates Alternative Autophagy Pathways and Mitochondrial Homeostasis.
    Elisa Teyssou, Laura Chartier, Delphine Roussel, Nirma D Perera, Ivan Nemazanyy, Dominique Langui, Mélanie Albert, Thierry Larmonier, Safaa Saker, François Salachas, Pierre-François Pradat, Vincent Meininger, Philippe Ravassard, Francine Côté, Christian S Lobsiger, Séverine Boillée, Bradley J Turner, Danielle Seilhean, Stéphanie Millecamps.
      Mutations in profilin 1 (PFN1) have been identified in rare familial cases of Amyotrophic Lateral Sclerosis (ALS). PFN1 is involved in multiple pathways that could intervene in ALS pathology. However, the specific pathogenic role of PFN1 mutations in ALS is still not fully understood. We hypothesized that PFN1 could play a role in regulating autophagy pathways and that PFN1 mutations could disrupt this function. We used patient cells (lymphoblasts) or tissue (post-mortem) carrying PFN1 mutations (M114T and E117G), and designed experimental models expressing wild-type or mutant PFN1 (cell lines and novel PFN1 mice established by lentiviral transgenesis) to study the effects of PFN1 mutations on autophagic pathway markers. We observed no accumulation of PFN1 in the spinal cord of one E117G mutation carrier. Moreover, in patient lymphoblasts and transfected cell lines, the M114T mutant PFN1 protein was unstable and deregulated the RAB9-mediated alternative autophagy pathway involved in the clearance of damaged mitochondria. In vivo, motor neurons expressing M114T mutant PFN1 showed mitochondrial abnormalities. Our results demonstrate that the M114T PFN1 mutation is more deleterious than the E117G variant in patient cells and experimental models and suggest a role for the RAB9-dependent autophagic pathway in ALS.
    Keywords:  ALS; NSC-34 cell line; RAB9; alternative autophagy; genetics; lymphoblasts; mitochondrial homeostasis; mutations; post-mortem spinal cord; transgenic mice
    DOI:  https://doi.org/10.3390/ijms23105694
  26. Int J Biol Macromol. 2022 May 23. pii: S0141-8130(22)01109-6. [Epub ahead of print]
    Autophagy protein LC3C binding to phospholipid and interaction with lipid membranes.
    Uxue Ballesteros, Asier Etxaniz, Marina N Iriondo, Yaiza R Varela, Melisa Lázaro, Ana R Viguera, L Ruth Montes, Mikel Valle, Félix M Goñi, Alicia Alonso.
      Autophagy is a process in which parts of the eukaryotic cell are selectively degraded in the lysosome. The materials to be catabolized are first surrounded by a double-membrane structure, the autophagosome. Autophagosome generation is a complex event, in which many proteins are involved. Among the latter, yeast Atg8 or its mammalian orthologues are essential in autophagosome membrane elongation, shaping and closure. A subfamily of the human Atg8 orthologues is formed by the proteins LC3A, LC3B, and LC3C. Previous studies suggest that, at variance with the other two, LC3C does not participate in cardiolipin-mediated mitophagy. The present study was devoted to exploring the binding of LC3C to lipid vesicles, bilayers and monolayers, and the ensuing protein-dependent perturbing effects, in the absence of the mitochondrial lipid cardiolipin. All Atg8 orthologues are covalently bound to a phospholipid prior to their involvement in autophagosome elongation. In our case, a mutant in the C-terminal amino acid, LC3C G126C, together with the use of a maleimide-derivatized phosphatidyl ethanolamine, ensured LC3C lipidation, up to 100% under certain conditions. Ultracentrifugation, surface pressure measurements, spectroscopic and cryo-electron microscopic techniques revealed that lipidated LC3C induced vesicle aggregation (5-fold faster in sonicated than in large unilamellar vesicles) and inter-vesicular lipid mixing (up to 82%), including inner-monolayer lipid mixing (up to 32%), consistent with in vitro partial vesicle fusion. LC3C was also able to cause the release of 80-90% vesicular aqueous contents. The data support the idea that LC3C would be able to help in autophagosome elongation/fusion in autophagy phenomena.
    Keywords:  Autophagy proteins; Lipid-protein interaction; Protein perturbation of membrane architecture
    DOI:  https://doi.org/10.1016/j.ijbiomac.2022.05.129
  27. Biomedicines. 2022 Apr 29. pii: 1027. [Epub ahead of print]10(5):
    Autophagy Modulation in Aggresome Formation: Emerging Implications and Treatments of Alzheimer's Disease.
    Md Ataur Rahman, M D Hasanur Rahman, A N M Mamun-Or-Rashid, Hongik Hwang, Sooyoung Chung, Bonglee Kim, Hyewhon Rhim.
      Alzheimer's disease (AD) is one of the most prevailing neurodegenerative diseases in the world, which is characterized by memory dysfunction and the formation of tau and amyloid β (Aβ) aggregates in multiple brain regions, including the hippocampus and cortex. The formation of senile plaques involving tau hyperphosphorylation, fibrillar Aβ, and neurofibrillary tangles (NFTs) is used as a pathological marker of AD and eventually produces aggregation or misfolded protein. Importantly, it has been found that the failure to degrade these aggregate-prone proteins leads to pathological consequences, such as synaptic impairment, cytotoxicity, neuronal atrophy, and memory deficits associated with AD. Recently, increasing evidence has suggested that the autophagy pathway plays a role as a central cellular protection system to prevent the toxicity induced by aggregation or misfolded proteins. Moreover, it has also been revealed that AD-related protein aggresomes could be selectively degraded by autophagosome and lysosomal fusion through the autophagy pathway, which is known as aggrephagy. Therefore, the regulation of autophagy serve as a useful approach to modulate the formation of aggresomes associated with AD. This review focuses on the recent improvements in the application of natural compounds and small molecules as a potential therapeutic approach for AD prevention and treatment via aggrephagy.
    Keywords:  Alzheimer’s disease (AD); aggregation; aggrephagy; aggresome; autophagosomes; autophagy
    DOI:  https://doi.org/10.3390/biomedicines10051027
  28. EMBO J. 2022 May 27. e110352
    A bacterial effector counteracts host autophagy by promoting degradation of an autophagy component.
    Jia Xuan Leong, Margot Raffeiner, Daniela Spinti, Gautier Langin, Mirita Franz-Wachtel, Andrew R Guzman, Jung-Gun Kim, Pooja Pandey, Alyona E Minina, Boris Macek, Anders Hafrén, Tolga O Bozkurt, Mary Beth Mudgett, Frederik Börnke, Daniel Hofius, Suayib Üstün.
      Beyond its role in cellular homeostasis, autophagy plays anti- and promicrobial roles in host-microbe interactions, both in animals and plants. One prominent role of antimicrobial autophagy is to degrade intracellular pathogens or microbial molecules, in a process termed xenophagy. Consequently, microbes evolved mechanisms to hijack or modulate autophagy to escape elimination. Although well-described in animals, the extent to which xenophagy contributes to plant-bacteria interactions remains unknown. Here, we provide evidence that Xanthomonas campestris pv. vesicatoria (Xcv) suppresses host autophagy by utilizing type-III effector XopL. XopL interacts with and degrades the autophagy component SH3P2 via its E3 ligase activity to promote infection. Intriguingly, XopL is targeted for degradation by defense-related selective autophagy mediated by NBR1/Joka2, revealing a complex antagonistic interplay between XopL and the host autophagy machinery. Our results implicate plant antimicrobial autophagy in the depletion of a bacterial virulence factor and unravel an unprecedented pathogen strategy to counteract defense-related autophagy in plant-bacteria interactions.
    Keywords:  autophagy; effectors; immunity; ubiquitination; xenophagy
    DOI:  https://doi.org/10.15252/embj.2021110352
  29. Genes (Basel). 2022 May 12. pii: 868. [Epub ahead of print]13(5):
    A New Zebrafish Model to Measure Neuronal α-Synuclein Clearance In Vivo.
    Ana Lopez, Alena Gorb, Nuno Palha, Angeleen Fleming, David C Rubinsztein.
      The accumulation and aggregation of α-synuclein (α-SYN) is a common characteristic of synucleinopathies, such as Parkinson's Disease (PD), Dementia with Lewy Bodies (DLB) or Multiple System Atrophy (MSA). Multiplications of the wildtype gene of α-SYN (SNCA) and most point mutations make α-SYN more aggregate-prone, and are associated with mitochondrial defects, trafficking obstruction, and impaired proteostasis, which contribute to elevated neuronal death. Here, we present new zebrafish models expressing either human wildtype (wt), or A53T mutant, α-SYN that recapitulate the above-mentioned hallmarks of synucleinopathies. The appropriate clearance of toxic α-SYN has been previously shown to play a key role in maintaining cell homeostasis and survival. However, the paucity of models to investigate α-SYN degradation in vivo limits our understanding of this process. Based on our recently described imaging method for measuring tau protein clearance in neurons in living zebrafish, we fused human SNCA to the photoconvertible protein Dendra2 which enabled analyses of wt and A53T α-SYN clearance kinetics in vivo. Moreover, these zebrafish models can be used to investigate the kinetics of α-SYN aggregation and to study the mechanisms, and potential new targets, controlling the clearance of both soluble and aggregated α-SYN.
    Keywords:  Parkinson’s disease; aggregation; autophagy; axonal transport; neurodegeneration; protein clearance; zebrafish disease model; α-synuclein
    DOI:  https://doi.org/10.3390/genes13050868
  30. Front Aging Neurosci. 2022 ;14 845330
    Regulation of Mitophagy by Sirtuin Family Proteins: A Vital Role in Aging and Age-Related Diseases.
    Wei Wan, Fuzhou Hua, Pu Fang, Chang Li, Fumou Deng, Shoulin Chen, Jun Ying, Xifeng Wang.
      Sirtuins are protein factors that can delay aging and alleviate age-related diseases through multiple molecular pathways, mainly by promoting DNA damage repair, delaying telomere shortening, and mediating the longevity effect of caloric restriction. In the last decade, sirtuins have also been suggested to exert mitochondrial quality control by mediating mitophagy, which targets damaged mitochondria and delivers them to lysosomes for degradation. This is especially significant for age-related diseases because dysfunctional mitochondria accumulate in aging organisms. Accordingly, it has been suggested that sirtuins and mitophagy have many common and interactive aspects in the aging process. This article reviews the mechanisms and pathways of sirtuin family-mediated mitophagy and further discusses its role in aging and age-related diseases.
    Keywords:  age-related disease; aging; mitochondria; mitophagy; neurodegenerative diseases; sirtuins
    DOI:  https://doi.org/10.3389/fnagi.2022.845330
  31. Gut. 2022 May 27. pii: gutjnl-2021-325117. [Epub ahead of print]
    Lysosomal lipid switch sensitises to nutrient deprivation and mTOR targeting in pancreatic cancer.
    Maria Chiara De Santis, Luca Gozzelino, Jean Piero Margaria, Andrea Costamagna, Edoardo Ratto, Federico Gulluni, Enza Di Gregorio, Erica Mina, Nicla Lorito, Marina Bacci, Rossano Lattanzio, Gianluca Sala, Paola Cappello, Francesco Novelli, Elisa Giovannetti, Caterina Vicentini, Silvia Andreani, Pietro Delfino, Vincenzo Corbo, Aldo Scarpa, Paolo Ettore Porporato, Andrea Morandi, Emilio Hirsch, Miriam Martini.
      OBJECTIVE: Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with limited therapeutic options. However, metabolic adaptation to the harsh PDAC environment can expose liabilities useful for therapy. Targeting the key metabolic regulator mechanistic target of rapamycin complex 1 (mTORC1) and its downstream pathway shows efficacy only in subsets of patients but gene modifiers maximising response remain to be identified.DESIGN: Three independent cohorts of PDAC patients were studied to correlate PI3K-C2γ protein abundance with disease outcome. Mechanisms were then studied in mouse (KPC mice) and cellular models of PDAC, in presence or absence of PI3K-C2γ (WT or KO). PI3K-C2γ-dependent metabolic rewiring and its impact on mTORC1 regulation were assessed in conditions of limiting glutamine availability. Finally, effects of a combination therapy targeting mTORC1 and glutamine metabolism were studied in WT and KO PDAC cells and preclinical models.
    RESULTS: PI3K-C2γ expression was reduced in about 30% of PDAC cases and was associated with an aggressive phenotype. Similarly, loss of PI3K-C2γ in KPC mice enhanced tumour development and progression. The increased aggressiveness of tumours lacking PI3K-C2γ correlated with hyperactivation of mTORC1 pathway and glutamine metabolism rewiring to support lipid synthesis. PI3K-C2γ-KO tumours failed to adapt to metabolic stress induced by glutamine depletion, resulting in cell death.
    CONCLUSION: Loss of PI3K-C2γ prevents mTOR inactivation and triggers tumour vulnerability to RAD001 (mTOR inhibitor) and BPTES/CB-839 (glutaminase inhibitors). Therefore, these results might open the way to personalised treatments in PDAC with PI3K-C2γ loss.
    Keywords:  AMINO ACIDS; CELL BIOLOGY; LIPID METABOLISM; PANCREATIC CANCER; SIGNAL TRANSDUCTION
    DOI:  https://doi.org/10.1136/gutjnl-2021-325117
  32. Autophagy. 2022 May 26. 1-3
    VMP1 regulates hepatic lipoprotein secretion and NASH independent of autophagy.
    Xiaoxiao Jiang, Allen Chen, Wen-Xing Ding, Hong-Min Ni.
      VMP1 is an ER membrane protein with phospholipid scramblase activity that has a critical role in regulating phagophore expansion and autophagosome closure. VMP1 also regulates lipid droplet formation and lipoprotein secretion in cultured cells and zebrafish. In a recent study, we showed that mice with hepatic deletion of Vmp1 have impaired very-low-density lipoprotein (VLDL) secretion and develop nonalcoholic steatohepatitis (NASH) even when fed with regular chow diet. Mechanistically, deletion of Vmp1 leads to decreased hepatic phosphatidylcholine (PC) and phosphatidylethanolamine (PE) levels as well as altered PC and PE acyl chain compositions resulting in the accumulation of neutral lipid structures in the ER phospholipid bilayer and decreased pre-VLDL assembly. These studies provide novel mechanistic insights into the non-autophagic functions of VMP1 in regulating lipoprotein secretion.
    Keywords:  ATG2; ER; TMEM41B; VLDL; VPS13D; autophagy; mitochondria
    DOI:  https://doi.org/10.1080/15548627.2022.2080958
  33. Acta Biochim Biophys Sin (Shanghai). 2022 Apr 25. 54(4): 524-536
    RETREG1-mediated ER-phagy activation induced by glucose deprivation alleviates nucleus pulposus cell damage via ER stress pathway.
    Rongjin Luo, Huaizhen Liang, Weifeng Zhang, Gaocai Li, Kangcheng Zhao, Wenbin Hua, Yu Song, Cao Yang.
      Accumulating evidence indicates that ER-phagy serves as a key adaptive regulatory mechanism in response to various stress conditions. However, the exact mechanisms underlying ER-phagy in the pathogenesis of intervertebral disc degeneration remain largely unclear. In the present study, we demonstrated that RETREG1-mediated ER-phagy is induced by glucose deprivation (GD) treatment, along with ER stress activation and cell function decline. Importantly, ER-phagy was shown to be crucial for cell survival under GD conditions. Furthermore, ER stress was suggested as an upstream event of ER-phagy upon GD treatment and upregulation of ER-phagy could counteract the ER stress response. Therefore, our findings indicate that RETREG1-mediated ER-phagy activation protects against GD treatment-induced cell injury via modulating ER stress in human nucleus pulposus cells.
    Keywords:  ER stress; ER-phagy; apoptosis; intervertebral disc degeneration; senescence
    DOI:  https://doi.org/10.3724/abbs.2022024
  34. Biomolecules. 2022 Apr 23. pii: 625. [Epub ahead of print]12(5):
    Nuclear and Cytoplasmatic Players in Mitochondria-Related CNS Disorders: Chromatin Modifications and Subcellular Trafficking.
    Matteo Gasparotto, Yi-Shin Lee, Alessandra Palazzi, Marcella Vacca, Francesco Filippini.
      Aberrant mitochondrial phenotypes are common to many central nervous system (CNS) disorders, including neurodegenerative and neurodevelopmental diseases. Mitochondrial function and homeostasis depend on proper control of several biological processes such as chromatin remodeling and transcriptional control, post-transcriptional events, vesicle and organelle subcellular trafficking, fusion, and morphogenesis. Mutation or impaired regulation of major players that orchestrate such processes can disrupt cellular and mitochondrial dynamics, contributing to neurological disorders. The first part of this review provides an overview of a functional relationship between chromatin players and mitochondria. Specifically, we relied on specific monogenic CNS disorders which share features with mitochondrial diseases. On the other hand, subcellular trafficking is coordinated directly or indirectly through evolutionarily conserved domains and proteins that regulate the dynamics of membrane compartments and organelles, including mitochondria. Among these "building blocks", longin domains and small GTPases are involved in autophagy and mitophagy, cell reshaping, and organelle fusion. Impairments in those processes significantly impact CNS as well and are discussed in the second part of the review. Hopefully, in filling the functional gap between the nucleus and cytoplasmic organelles new routes for therapy could be disclosed.
    Keywords:  MeCP2; Rab; SNARE; VAMP; autophagy; chromatin remodeling; longin domain; mitochondria; mitophagy; small GTPase; subcellular trafficking
    DOI:  https://doi.org/10.3390/biom12050625
  35. Int J Mol Sci. 2022 May 10. pii: 5311. [Epub ahead of print]23(10):
    MoWhi2 Mediates Mitophagy to Regulate Conidiation and Pathogenesis in Magnaporthe oryzae.
    Shuai Meng, Jane Sadhna Jagernath, Chaoxi Luo, Huanbin Shi, Yanjun Kou.
      Mitophagy refers to the specific process of degrading mitochondria, which is an important physiological process to maintain the balance of mitochondrial quantity and quality in cells. At present, the mechanisms of mitophagy in pathogenic fungi remain unclear. Magnaporthe oryzae (Syn. Pyricularia oryzae), the causal agent of rice blast disease, is responsible for the most serious disease of rice. In M. oryzae, mitophagy occurs in the foot cells and invasive hyphae to promote conidiation and infection. In this study, fluorescent observations and immunoblot analyses showed that general stress response protein MoWhi2 is required for mitophagy in M. oryzae. In addition, the activation of the autophagy, pexophagy and cytoplasm-to-vacuole targeting (CVT) pathway upon nitrogen starvation was determined using the GFP-MoATG8, GFP-SRL and MoAPE1-GFP strains and the ΔMowhi2 mutant in these backgrounds. The results indicated that MoWhi2 is specifically required for mitophagy in M. oryzae. Further studies showed that mitophagy in the foot cells and invasive hyphae of the ΔMowhi2 was interrupted, leading to reduced conidiation and virulence in the ΔMowhi2 mutant. Taken together, we found that MoWhi2 contributes to conidiation and invasive growth by regulating mitophagy in M. oryzae.
    Keywords:  Whi2; mitophagy; pathogenesis; rice blast
    DOI:  https://doi.org/10.3390/ijms23105311
  36. Antioxidants (Basel). 2022 May 20. pii: 1009. [Epub ahead of print]11(5):
    Diclofenac: A Nonsteroidal Anti-Inflammatory Drug Inducing Cancer Cell Death by Inhibiting Microtubule Polymerization and Autophagy Flux.
    Soohee Choi, Suree Kim, Jiyoung Park, Seung Eun Lee, Chaewon Kim, Dongmin Kang.
      Diclofenac, a nonsteroidal anti-inflammatory drug (NSAID) used to treat inflammatory diseases induces cellular toxicity by increasing the production of reactive oxygen species (ROS) and impairing autophagic flux. In this study, we investigated whether diclofenac induces cancer cell death and the mechanism by which diclofenac causes cell death. We observed that diclofenac induces mitotic arrest with a half-maximal effective concentration of 170 μM and cell death with a half-maximal lethal dose of 200 µM during 18-h incubation in HeLa cells. Cellular microtubule imaging and in vitro tubulin polymerization assays demonstrated that treatment with diclofenac elicits microtubule destabilization. Autophagy relies on microtubule-mediated transport and the fusion of autophagic vesicles. We observed that diclofenac inhibits both phagophore movement, an early step of autophagy, and the fusion of autophagosomes and lysosomes, a late step of autophagy. Diclofenac also induces the fragmentation of mitochondria and the Golgi during cell death. We found that diclofenac induces cell death further in combination with 5-fuorouracil, a DNA replication inhibitor than in single treatment in cancer cells. Pancreatic cancer cells, which have high basal autophagy, are particularly sensitive to cell death by diclofenac. Our study suggests that microtubule destabilization by diclofenac induces cancer cell death via compromised spindle assembly checkpoints and increased ROS through impaired autophagy flux. Diclofenac may be a candidate therapeutic drug in certain type of cancers by inhibiting microtubule-mediated cellular events in combination with clinically utilized nucleoside metabolic inhibitors, including 5-fluorouracil, to block cancer cell proliferation.
    Keywords:  autophagy; cell death; combination cancer therapy; diclofenac; microtubule depolymerization
    DOI:  https://doi.org/10.3390/antiox11051009
  37. Toxins (Basel). 2022 May 22. pii: 360. [Epub ahead of print]14(5):
    Modulation of Ricin Intoxication by the Autophagy Inhibitor EACC.
    Kirsten Sandvig, Simona Kavaliauskiene, Anne Grethe Myrann, Tore Geir Iversen, Tore Skotland.
      The compound EACC (ethyl (2-(5-nitrothiophene-2-carboxamido) thiophene-3-carbonyl) carbamate) was recently reported to inhibit fusion of autophagosomes with lysosomes in a reversible manner by inhibiting recruitment of syntaxin 17 to autophagosomes. We report here that this compound also provides a strong protection against the protein toxin ricin as well as against other plant toxins such as abrin and modeccin. The protection did not seem to be caused by inhibition of endocytosis and retrograde transport, but rather by inhibited release of the enzymatically active A-moiety to the cytosol. The TANK-binding kinase 1 (TBK1) has been reported to phosphorylate syntaxin 17 and be required for initiation of autophagy. The inhibitor of TBK1, MRT68601, induced in itself a strong sensitization to ricin, apparently by increasing transport to the Golgi apparatus. Importantly, MRT68601 increased Golgi transport of ricin even in the presence of EACC, but EACC was still able to inhibit intoxication, supporting the idea that EACC protects at a late step along the retrograde pathway. These results also indicate that phosphorylation of syntaxin 17 is not required for the protection observed.
    Keywords:  Golgi apparatus; abrin; autophagy; endoplasmic reticulum; modeccin; plant toxins; retrograde transport; ricin; viscumin; volkensin
    DOI:  https://doi.org/10.3390/toxins14050360
  38. iScience. 2022 May 20. 25(5): 104334
    Increased glycine contributes to synaptic dysfunction and early mortality in Nprl2 seizure model.
    Brianne Dentel, Lidiette Angeles-Perez, Chongyu Ren, Vikram Jakkamsetti, Andrew J Holley, Daniel Caballero, Emily Oh, Jay Gibson, Juan M Pascual, Kimberly M Huber, Benjamin P Tu, Peter T Tsai.
      Targeted therapies for epilepsies associated with the mTORC1 signaling negative regulator GATOR1 are lacking. NPRL2 is a subunit of the GATOR1 complex and mutations in GATOR1 subunits, including NPRL2, are associated with epilepsy. To delineate the mechanisms underlying NPRL2-related epilepsies, we created a mouse (Mus musculus) model with neocortical loss of Nprl2. Mutant mice have increased mTORC1 signaling and exhibit spontaneous seizures. They also display abnormal synaptic function characterized by increased evoked and spontaneous EPSC and decreased evoked and spontaneous IPSC frequencies, respectively. Proteomic and metabolomics studies of Nprl2 mutants revealed alterations in known epilepsy-implicated proteins and metabolic pathways, including increases in the neurotransmitter, glycine. Furthermore, glycine actions on the NMDA receptor contribute to the electrophysiological and survival phenotypes of these mice. Taken together, in this neuronal Nprl2 model, we delineate underlying molecular, metabolic, and electrophysiological mechanisms contributing to mTORC1-related epilepsy, providing potential therapeutic targets for epilepsy.
    Keywords:  metabolomics; molecular neuroscience; neuroscience; omics
    DOI:  https://doi.org/10.1016/j.isci.2022.104334
  39. Eur J Cell Biol. 2022 May 19. pii: S0171-9335(22)00043-7. [Epub ahead of print]101(3): 151240
    Amino acids control blood glucose levels through mTOR signaling.
    Jialin Fan, Ziqiang Yuan, Stephen K Burley, Steven K Libutti, X F Steven Zheng.
      Amino Acids are not only major nutrient sources, but also serve as chemical signals to control cellular growth. Rab1A recently emerged as a key component in amino acid sensing and signaling to activate the mTOR complex1 (mTORC1). In a recently published study [1], we generated tamoxifen-inducible, conditional whole-body Rab1A knockout in adult mice. These mice are viable but develop hyperglycemia and glucose intolerance. Interestingly, Rab1A ablation selectively reduces insulin expression and pancreatic beta-cell population. Mechanistically, branched chain amino acids (BCAA), through the Rab1A-mTORC1 complex, promote the stability and nuclear localization of Pdx1, a master transcription factor that controls growth, function and identity of pancreatic beta-cells. These findings reveal a role and underlying mechanism by which amino acids control body's glucose level through a beta-cell specific function by the Rab1A-mTORC1-Pdx1 signaling axis, which has implications in both diabetes and cancer.
    Keywords:  Alpha cell; Amino acid; Beta cell; Cancer; Diabetes; Insulin; Islet; MTOR; Pancreas; Rab1A
    DOI:  https://doi.org/10.1016/j.ejcb.2022.151240
  40. Cell Death Differ. 2022 May 25.
    Plin2-mediated lipid droplet mobilization accelerates exit from pluripotency by lipidomic remodeling and histone acetylation.
    Yi Wu, Keshi Chen, Linpeng Li, Zhihong Hao, Tianyu Wang, Yang Liu, Guangsuo Xing, Zichao Liu, Heying Li, Hao Yuan, Jianghuan Lu, Cheng Zhang, Jinye Zhang, Danyun Zhao, Junwei Wang, Jinfu Nie, Dan Ye, Guangjin Pan, Wai-Yee Chan, Xingguo Liu.
      Metabolic switch is critical for cell fate determination through metabolic functions, epigenetic modifications, and gene expression. However, the mechanisms underlying these alterations and their functional roles remain unclear. Here, we show that Plin2-mediated moderate lipid hydrolysis is critical for pluripotency of embryonic stem cells (ESCs). Upon exit from pluripotency, lipid droplet (LD)-associated protein Plin2 is recognized by Hsc70 and degraded via chaperone-mediated autophagy to facilitate LD mobilization. Enhancing lipid hydrolysis by Plin2 knockout promotes pluripotency exit, which is recovered by ATGL inhibition. Mechanistically, excessive lipid hydrolysis induces a dramatic lipidomic remodeling characterized by decreased cardiolipin and phosphatidylethanolamine, which triggers defects in mitochondrial cristae and fatty acid oxidation, resulting in reduced acetyl-CoA and histone acetylation. Our results reveal how LD mobilization is regulated and its critical role in ESC pluripotency, and indicate the mechanism linking LD homeostasis to mitochondrial remodeling and epigenetic regulation, which might shed light on development and diseases.
    DOI:  https://doi.org/10.1038/s41418-022-01018-8
  41. Oxid Med Cell Longev. 2022 ;2022 4759963
    Mechanisms of Mitochondrial Malfunction in Alzheimer's Disease: New Therapeutic Hope.
    Showkat Ul Nabi, Andleeb Khan, Ehraz Mehmood Siddiqui, Muneeb U Rehman, Saeed Alshahrani, Azher Arafah, Sidharth Mehan, Rana M Alsaffar, Athanasios Alexiou, Bairong Shen.
      Mitochondria play a critical role in neuron viability or death as it regulates energy metabolism and cell death pathways. They are essential for cellular energy metabolism, reactive oxygen species production, apoptosis, Ca++ homeostasis, aging, and regeneration. Mitophagy and mitochondrial dynamics are thus essential processes in the quality control of mitochondria. Improvements in several fundamental features of mitochondrial biology in susceptible neurons of AD brains and the putative underlying mechanisms of such changes have made significant progress. AD's etiology has been reported by mitochondrial malfunction and oxidative damage. According to several recent articles, a continual fusion and fission balance of mitochondria is vital in their normal function maintenance. As a result, the shape and function of mitochondria are inextricably linked. This study examines evidence suggesting that mitochondrial dysfunction plays a significant early impact on AD pathology. Furthermore, the dynamics and roles of mitochondria are discussed with the link between mitochondrial malfunction and autophagy in AD has also been explored. In addition, recent research on mitochondrial dynamics and mitophagy in AD is also discussed in this review. It also goes into how these flaws affect mitochondrial quality control. Furthermore, advanced therapy techniques and lifestyle adjustments that lead to improved management of the dynamics have been demonstrated, hence improving the conditions that contribute to mitochondrial dysfunction in AD.
    DOI:  https://doi.org/10.1155/2022/4759963
  42. Anim Nutr. 2022 Sep;10 12-18
    The ubiquitin ligase Nedd4-2 mediates the regulation of PepT2 by mTORC1 in bovine mammary epithelial cells.
    Caihong Wang, Fengqi Zhao, Jianxin Liu, Hongyun Liu.
      Peptide transporter 2 (PepT2) transports short peptides from the blood into bovine mammary epithelial cells (BMEC) to stimulate milk protein synthesis. Despite the fact that the effect of PepT2 is acknowledged in BMEC, little is known about its regulation. This study was completed to investigate the role of mammalian target of the rapamycin (mTOR) signaling in regulating the expression and function of PepT2 in BMEC. The regulation of PepT2 by mTOR in BMEC was studied in vitro using peptide transport assay, gene silencing, Western blot. The membrane expression of PepT2 and the uptake of β-Ala-Lys-N-7-amino-4-methylcoumarin-3-acetic acid (β-Ala-Lys-AMCA), a model dipeptide, in BMEC were reduced by rapamycin (a mTOR inhibitor) and silencing of either mTOR complex 1 (mTORC1) or mTOR complex 2 (mTORC2), stimulated by DEP domain-containing mTOR-interacting protein (DEPTOR, endogenous inhibitor of mTORC1 and mTORC2) silencing. The trafficking of PepT2 to the membrane and the uptake of β-Ala-Lys-AMCA was promoted by neuronal precursor cell-expressed developmentally down-regulated 4 isoform 2 (Nedd4-2) silencing. The effects of knockdown of mTORC1, but not mTORC2, on cell membrane expression and transport activity of PepT2 was abolished by Nedd4-2 silencing. With immunofluorescence staining, PepT2 was identified to be interacting with Nedd4-2. The Nedd4-2 expression and the interaction between PepT2 and Nedd4-2 was increased through mTORC1 knockdown, indicating an increased ubiquitination of PepT2. The results revealed that mTORC1 can regulate the expression and function of PepT2 through Nedd4-2 in BMEC.
    Keywords:  BMEC; Nedd4-2; PepT2; Peptide uptake; mTORC1; mTORC2
    DOI:  https://doi.org/10.1016/j.aninu.2021.11.008
  43. Biomolecules. 2022 Apr 21. pii: 614. [Epub ahead of print]12(5):
    Roles of mTOR in the Regulation of Pancreatic β-Cell Mass and Insulin Secretion.
    Shun-Ichiro Asahara, Hiroyuki Inoue, Hitoshi Watanabe, Yoshiaki Kido.
      Pancreatic β-cells are the only type of cells that can control glycemic levels via insulin secretion. Thus, to explore the mechanisms underlying pancreatic β-cell failure, many reports have clarified the roles of important molecules, such as the mechanistic target of rapamycin (mTOR), which is a central regulator of metabolic and nutrient cues. Studies have uncovered the roles of mTOR in the function of β-cells and the progression of diabetes, and they suggest that mTOR has both positive and negative effects on pancreatic β-cells in the development of diabetes.
    Keywords:  ER stress; autophagy; insulin secretion; mTOR; pancreatic β-cell
    DOI:  https://doi.org/10.3390/biom12050614
  44. Parkinsons Dis. 2022 ;2022 6600944
    Association of p53 with Neurodegeneration in Parkinson's Disease.
    Qiang Luo, Wei Sun, Yi-Fan Wang, Ji Li, Da-Wei Li.
      p53 is a vital transcriptional protein implicated in regulating diverse cellular processes, including cell cycle arrest, DNA repair, mitochondrial metabolism, redox homeostasis, autophagy, senescence, and apoptosis. Recent studies have revealed that p53 levels and activity are substantially increased in affected neurons in cellular and animal models of Parkinson's disease (PD) as well as in the brains of PD patients. p53 activation in response to neurodegenerative stress is closely associated with the degeneration of dopaminergic neurons accompanied by mitochondrial dysfunction, reactive oxygen species (ROS) production, abnormal protein aggregation, and impairment of autophagy, and these pathogenic events have been implicated in the pathogenesis of PD. Pathogenic p53 integrates diverse cellular stresses and activate these downstream events to induce the degeneration of dopaminergic neurons; thus, it plays a crucial role in the pathogenesis of PD and appears to be a potential target for the treatment of the disease. We reviewed the current knowledge concerning p53-dependent neurodegeneration to better understand the underlying mechanisms and provide possible strategies for PD treatment by targeting p53.
    DOI:  https://doi.org/10.1155/2022/6600944
  45. Sci Total Environ. 2022 May 24. pii: S0048-9697(22)03315-0. [Epub ahead of print] 156218
    N, N-dimethylformamide exposure induced liver abnormal mitophagy by targeting miR-92a-1-5p-BNIP3L pathway in vivo and vitro.
    Lin Xu, Wanli Ma, Yuan Jin, Xueying Sun, Ningning Chen, Xiaoxiao Zhu, Jiao Luo, Chuanhai Li, Kunming Zhao, Yuxin Zheng, Dianke Yu.
      N, N-dimethylformamide (DMF) is a widely existing harmful environmental pollutant from industrial emission which can threat human health for both occupational and general populations. Epidemiological and experimental studies have indicated liver as the primary target organ of DMF. However, the molecular mechanism under DMF-induced hepatoxicity remains unclear. In the present study, we identified that DMF could induce abnormal autophagy flux in cells. We also showed that DMF-induced mitochondrial dysfunction and lethal mitophagy which further leads to autophagic cell death. Next, miRNA microarray analysis identified miR-92a-1-5p as the most down-regulated miRNA upon DMF exposure. Mechanistically, miR-92a-1-5p regulated mitochondrial function and mitophagy by targeting mitochondrial protein BNIP3L. Exogenous miR-92a-1-5p significantly attenuated DMF-induced mitochondrial dysfunction and mitophagy in vitro and in vivo. Our study highlights the mechanistic link between miRNAs and mitophagy under environmental stress, which provided a new clue for the mitochondrial epigenetics mechanism on environmental toxicant-induced hepatoxicity.
    Keywords:  Hepatoxicity; Mitochondrial dysfunction; Mitophagy; N; N-dimethylformamide; miR-92a-1-5p
    DOI:  https://doi.org/10.1016/j.scitotenv.2022.156218
  46. J Neuroinflammation. 2022 May 22. 19(1): 113
    A breakdown in microglial metabolic reprogramming causes internalization dysfunction of α-synuclein in a mouse model of Parkinson's disease.
    Jia Lu, Chenfei Wang, Xin Cheng, Ruizhi Wang, Xuehan Yan, Pengju He, Hongzhuan Chen, Zhihua Yu.
      BACKGROUND: The α-synuclein released by neurons activates microglia, which then engulfs α-synuclein for degradation via autophagy. Reactive microglia are a major pathological feature of Parkinson's disease (PD), although the exact role of microglia in the pathogenesis of PD remains unclear. Transient receptor potential vanilloid type 1 (TRPV1) channels are nonselective cation channel protein that have been proposed as neuroprotective targets in neurodegenerative diseases.METHODS: Using metabolic profiling, microglia energy metabolism was measured including oxidative phosphorylation and aerobic glycolysis. The mRFP-GFP-tagged LC3 reporter was introduced to characterize the role of TRPV1 in microglial autophagy. α-synuclein preformed fibril (PFF) TRPV1flox/flox; Cx3cr1Cre mouse model of sporadic PD were employed to study the capacity of TRPV1 activation to attenuate neurodegeneration process.
    RESULTS: We found that acute exposure to PFF caused microglial activation as a result of metabolic reprogramming from oxidative phosphorylation to aerobic glycolysis via the AKT-mTOR-HIF-1α pathway. Activated microglia eventually reached a state of chronic PFF-tolerance, accompanied by broad defects in energy metabolism. We showed that metabolic boosting by treatment with the TRPV1 agonist capsaicin rescued metabolic impairments in PFF-tolerant microglia and also defects in mitophagy caused by disruption of the AKT-mTOR-HIF-1α pathway. Capsaicin attenuated phosphorylation of α-synuclein in primary neurons by boosting phagocytosis in PFF-tolerant microglia in vitro. Finally, we found that behavioral deficits and loss of dopaminergic neurons were accelerated in the PFF TRPV1flox/flox; Cx3cr1Cre mouse model of sporadic PD. We identified defects in energy metabolism, mitophagy and phagocytosis of PFF in microglia from the substantia nigra pars compacta of TRPV1flox/flox; Cx3cr1Cre mice.
    CONCLUSION: The findings suggest that modulating microglial metabolism might be a new therapeutic strategy for PD.
    Keywords:  Autophagy; Capsaicin; Microglia; TRPV1; α-Synuclein
    DOI:  https://doi.org/10.1186/s12974-022-02484-0
  47. J Mol Neurosci. 2022 May 27.
    Impact of Raptor and Rictor Deletion on Hippocampal Pathology Following Status Epilepticus.
    Christin M Godale, Emma V Parkins, Christina Gross, Steve C Danzer.
      Neuronal hyperactivation of the mTOR signaling pathway may play a role in driving the pathological sequelae that follow status epilepticus. Animal studies using pharmacological tools provide support for this hypothesis, however, systemic inhibition of mTOR-a growth pathway active in every mammalian cell-limits conclusions on cell type specificity. To circumvent the limitations of pharmacological approaches, we developed a viral/genetic strategy to delete Raptor or Rictor, inhibiting mTORC1 or mTORC2, respectively, from excitatory hippocampal neurons after status epilepticus in mice. Raptor or Rictor was deleted from roughly 25% of hippocampal granule cells, with variable involvement of other hippocampal neurons, after pilocarpine status epilepticus. Status epilepticus induced the expected loss of hilar neurons, sprouting of granule cell mossy fiber axons and reduced c-Fos activation. Gene deletion did not prevent these changes, although Raptor loss reduced the density of c-Fos-positive granule cells overall relative to Rictor groups. Findings demonstrate that mTOR signaling can be effectively modulated with this approach and further reveal that blocking mTOR signaling in a minority (25%) of granule cells is not sufficient to alter key measures of status epilepticus-induced pathology. The approach is suitable for producing higher deletion rates, and altering the timing of deletion, which may lead to different outcomes.
    Keywords:  Dentate granule cells; Temporal lobe epilepsy; c-Fos; mTOR; mTORC1; mTORC2
    DOI:  https://doi.org/10.1007/s12031-022-02030-w
  48. Molecules. 2022 May 13. pii: 3142. [Epub ahead of print]27(10):
    Infection Dynamics of ATG8 in Leishmania: Balancing Autophagy for Therapeutics.
    Vrushali Guhe, Farah Anjum, Alaa Shafie, Md Imtaiyaz Hassan, Visweswara Rao Pasupuleti, Shailza Singh.
      In many regions of the world, Leishmaniasis is a cause of substantial mortality and ailment. Due to impediment in available treatment, development of novel and effective treatments is indispensable. Significance of autophagy has been accentuated in infectious disease as well as in Leishmaniasis, and it is having capability to be manifested as a therapeutic target. By evincing autophagy as a novel therapeutic regime, this study emphasized on the critical role of ATG4.1-ATG8 and ATG5-ATG12 complexes in Leishmania species. The objective here was to identify ATG8 as a potential therapeutic target in Leishmania. R71T, P56E, R18P are the significant mutations which shows detrimental effect on ATG8 while Arg276, Arg73, Cys75 of ATG4.1 and Val88, Pro89, Glu116, Asn117, and Gly120 are interacting residues of ATG8. Along with this, we also bring into spotlight an enticing role of Thiabendazole derivatives that interferes with the survival mechanisms by targeting ATG8. Further, the study claims that thiabendazole can be a potential drug candidate to target autophagy process in the infectious disease Leishmaniasis.
    Keywords:  ATG proteins; Leishmaniasis; MD simulations; SCA; conservation of ATG proteins; docking; mutational studies; phylogeny; thiabendazole
    DOI:  https://doi.org/10.3390/molecules27103142
  49. Front Cell Dev Biol. 2022 ;10 906453
    Safeguarding Lysosomal Homeostasis by DNAJC5/CSPα-Mediated Unconventional Protein Secretion and Endosomal Microautophagy.
    Juhyung Lee, Yue Xu, Yihong Ye.
      Neuronal ceroid lipofuscinosis (NCL) is a collection of genetically inherited neurological disorders characterized by vision loss, seizure, brain death, and premature lethality. At the cellular level, a key pathologic hallmark of NCL is the build-up of autofluorescent storage materials (AFSM) in lysosomes of both neurons and non-neuronal cells. Molecular dissection of the genetic lesions underlying NCLs has shed significant insights into how disruption of lysosomal homeostasis may lead to lipofuscin accumulation and NCLs. Intriguingly, recent studies on DNAJC5/CSPα, a membrane associated HSC70 co-chaperone, have unexpectedly linked lipofuscin accumulation to two intimately coupled protein quality control processes at endolysosomes. This review discusses how deregulation of unconventional protein secretion and endosomal microautophagy (eMI) contributes to lipofuscin accumulation and neurodegeneration.
    Keywords:  DNAJC5/CSPα; ceroid lipofuscinosis neuronal; cysteine string protein; endosomal microautophagy; lysosome; misfolding-associated protein secretion (MAPS); protein quality control; unconventional protein secretion
    DOI:  https://doi.org/10.3389/fcell.2022.906453
  50. Neuroscience. 2022 May 18. pii: S0306-4522(22)00250-0. [Epub ahead of print]
    Autophagy:a new mechanism for esketamine as a depression therapeutic.
    Guanghao Jiang, Yibo Wang, Qingzhen Liu, Tingting Gu, Suting Liu, Anqi Yin, Lidong Zhang.
      Depression is a serious physical and mental disease, with major depressive disorder (MDD) being a hard-to-treat, life-threatening form of the condition. Currently, esketamine (ESK) is used in the clinical treatment of MDD, but the drug mechanisms continue to be unclear. In this study, we explored the therapeutic efficacy of ESK against lipopolysaccharide (LPS)-induced neuroinflammatory, autophagic, and depressive symptoms and the possible mechanisms behind them. Our study demonstrated that LPS increased cytokine levels (TNF-α, IL-1β, IL-6), induced neuroinflammation, led to increased levels of autophagy markers, and enhanced autophagy activation, which ultimately caused depressive symptoms in mouse models. ESK inhibited autophagy via the mTOR-BDNF signaling pathway and significantly alleviated the adverse effects induced by LPS, mainly in the form of reduced levels of cytokines, apoptotic factors, and autophagic markers; elevated BDNF levels; and improved depression-like behavior. Furthermore, we were interested to know if ESK in combination with other autophagy inhibitors would have a better antidepressant effect, and we chose the autophagy inhibitor 3-MA for this attempt. Interestingly, the use of 3-MA did not attenuate or even enhance the therapeutic effect of ESK. The results suggest that, in the LPS-induced depression models, ESK conveyed an antidepressant effect via the inhibition of autophagy through the mTOR-BDNF pathway.
    Keywords:  Autophagy; Depression; Esketamine; Lipopolysaccharides; mTOR
    DOI:  https://doi.org/10.1016/j.neuroscience.2022.05.014
  51. Brain. 2022 May 25. pii: awac191. [Epub ahead of print]
    Early bioenergetic and autophagy impairments at the Parkinson's disease synapse.
    G Aleph Prieto, Carl W Cotman.
      
    DOI:  https://doi.org/10.1093/brain/awac191
  52. iScience. 2022 Jun 17. 25(6): 104382
    A meiotic switch in lysosome activity supports spermatocyte development in young flies but collapses with age.
    Tyler J Butsch, Olga Dubuisson, Alyssa E Johnson, K Adam Bohnert.
      Gamete development ultimately influences animal fertility. Identifying mechanisms that direct gametogenesis, and how they deteriorate with age, may inform ways to combat infertility. Recently, we found that lysosomes acidify during oocyte maturation in Caenorhabditis elegans, suggesting that a meiotic switch in lysosome activity promotes female germ-cell health. Using Drosophila melanogaster, we report that lysosomes likewise acidify in male germ cells during meiosis. Inhibiting lysosomes in young-male testes causes E-cadherin accumulation and loss of germ-cell partitioning membranes. Notably, analogous changes occur naturally during aging; in older testes, a reduction in lysosome acidity precedes E-cadherin accumulation and membrane dissolution, suggesting one potential cause of age-related spermatocyte abnormalities. Consistent with lysosomes governing the production of mature sperm, germ cells with homozygous-null mutations in lysosome-acidifying machinery fail to survive through meiosis. Thus, lysosome activation is entrained to meiotic progression in developing sperm, as in oocytes, and lysosomal dysfunction may instigate male reproductive aging.
    Keywords:  Biological sciences; Cell biology; Developmental biology; Molecular biology
    DOI:  https://doi.org/10.1016/j.isci.2022.104382
  53. J Cell Biochem. 2022 May 23.
    Targeting the lysosome: Mechanisms and treatments for nonalcoholic fatty liver disease.
    Jing Pu.
      The multiple functions of the lysosome, including degradation, nutrient sensing, signaling, and gene regulation, enable the lysosome to regulate lipid metabolism at different levels. In this review, I summarize the recent studies on lysosomal regulation of lipid metabolism and the alterations of the lysosome functions in the livers affected by nonalcoholic fatty liver disease (NAFLD). NAFLD is a highly prevalent lipid metabolic disorder. The progression of NAFLD leads to nonalcoholic steatohepatitis (NASH) and other severe liver diseases, and thus the prevention and treatments of NAFLD progression are critically needed. Targeting the lysosome is a promising strategy. I also discuss the current manipulations of the lysosome functions in the preclinical studies of NAFLD and propose my perspectives on potential future directions.
    Keywords:  lipid droplet; lipid transport; lipotoxicity; lysosome dynamics; mTOR
    DOI:  https://doi.org/10.1002/jcb.30274
  54. J Viral Hepat. 2022 May 27.
    TRPM2 regulates autophagy to participate in hepatitis B virus replication.
    Liyan Chen, Liying Zhu, Xingyu Lu, Xiaoyu Ming, Baoshan Yang.
      Hepatitis B virus (HBV) affects over 300 million people across the world and is further associated with the self-digesting process of autophagy. Accordingly, the current study set out to explore the role of transient receptor potential cation channel subfamily M member 2 (TRPM2) in HBV replication. Firstly, Huh-7 cells were transfected with the pHBV1.3 plasmid to detect the expression patterns of TRPM2 and neutrophil cytosolic factor 1 (p47 phox), followed by evaluating the role of TRPM2 in autophagy and HBV replication and exploring the interaction between TRPM2 and p47 phox. Collaborative experiments were further designed to explore the role of p47 phox and autophagy in TRPM2 regulation of HBV replication, in addition to animal experimentation to validate the role of TRPM2/p47 phox axis in vivo. It was found that TRPM2 up-regulation was associated with HBV replication. On the other hand, silencing of TRPM2 inhibited HBV replication and autophagy in vitro and in vivo, as evidenced by reduced HBV DNA load, HBV mRNA, HBeAg, and HBsAg, and diminished autophagic spot number, LC3 II/I ratio, Beclin-1 expressions and increased p62 expressions. Mechanistic experimentation illustrated that TRPM2 interacted with p47 phox and positively regulated p47 phox, such that p47 phox up-regulation or use of Rapamycin (autophagy activator) weakened the inhibitory role of silencing TRPM2. Collectively, our findings indicated that HBV infection promotes TRPM2 expression, and TRPM2 interacts with p47 phox to induce autophagy and facilitate HVB replication.
    Keywords:  Autophagy; Hepatitis B virus; Protein-protein interaction; TRPM2; Viral replication; p47 phox
    DOI:  https://doi.org/10.1111/jvh.13710
  55. Front Oncol. 2022 ;12 893396
    Loss of PTEN-Induced Kinase 1 Regulates Oncogenic Ras-Driven Tumor Growth By Inhibiting Mitochondrial Fission.
    Dantong Zhu, Fengtong Han, Liuke Sun, Sandeep K Agnihotri, Ying Hu, Hansruedi Büeler.
      Mitochondrial metabolism and dynamics (fission and fusion) critically regulate cell survival and proliferation, and abnormalities in these pathways are implicated in both neurodegenerative disorders and cancer. Mitochondrial fission is necessary for the growth of mutant Ras-dependent tumors. Here, we investigated whether loss of PTEN-induced kinase 1 (PINK1) - a mitochondrial kinase linked to recessive familial Parkinsonism - affects the growth of oncogenic Ras-induced tumor growth in vitro and in vivo. We show that RasG12D-transformed embryonic fibroblasts (MEFs) from PINK1-deficient mice display reduced growth in soft agar and in nude mice, as well as increased necrosis and decreased cell cycle progression, compared to RasG12D-transformed MEFs derived from wildtype mice. PINK1 re-expression (overexpression) at least partially rescues these phenotypes. Neither PINK1 deletion nor PINK1 overexpression altered Ras expression levels. Intriguingly, PINK1-deficient Ras-transformed MEFs exhibited elongated mitochondria and altered DRP1 phosphorylation, a key event in regulating mitochondrial fission. Inhibition of DRP1 diminished PINK1-regulated mitochondria morphological changes and tumor growth suggesting that PINK1 deficiency primarily inhibits Ras-driven tumor growth through disturbances in mitochondrial fission and associated cell necrosis and cell cycle defects. Moreover, we substantiate the requirement of PINK1 for optimal growth of Ras-transformed cells by showing that human HCT116 colon carcinoma cells (carrying an endogenous RasG13D mutation) with CRISPR/Cas9-introduced PINK1 gene deletions also show reduced mitochondrial fission and decreased growth. Our results support the importance of mitochondrial function and dynamics in regulating the growth of Ras-dependent tumor cells and provide insight into possible mechanisms underlying the lower incidence of cancers in Parkinson's disease and other neurodegenerative disorders.
    Keywords:  PTEN-induced kinase-1 (PINK1); Ras protein; Ras-induced tumors; cell cycle; dynamin-related protein 1 (DRP1); mitochondrial dynamics; mitochondrial metabolism
    DOI:  https://doi.org/10.3389/fonc.2022.893396
  56. Int J Mol Sci. 2022 May 19. pii: 5697. [Epub ahead of print]23(10):
    An Experimental Study Reveals the Protective Effect of Autophagy against Realgar-Induced Liver Injury via Suppressing ROS-Mediated NLRP3 Inflammasome Pathway.
    Jing Yang, Jian Li, Haoqi Guo, Yuwei Zhang, Ziwei Guo, Yu Liu, Taoguang Huo.
      Realgar, a poisonous traditional Chinese medicine, has been shown to cause liver injury when used for long periods or overdoses. However, the underlying molecular mechanisms and therapeutic targets have not been fully elucidated. The aim of this study is to explore the role of autophagy in sub-chronic realgar exposure-induced liver injury. Here, the liver injury model was established by continuously administrating mice with 1.35 g/kg realgar for 8 weeks. 3-methyladenine (3-MA) and rapamycin (RAPA) were used to regulate autophagy. The results showed that realgar induced abnormal changes in liver function, pathological morphology, expression of inflammatory cytokines, and upregulated NLRP3 inflammasome pathway in mouse livers. RAPA treatment (an inducer of autophagy) significantly improved realgar-induced liver injury and NLRP3 inflammasome activation, while 3-MA (an inhibitor of autophagy) aggravated the realgar-induced liver injury and NLRP3 inflammasome activation. Furthermore, we found that realgar-induced NLRP3 inflammasome activation in mouse livers is mediated by ROS. RAPA eliminates excessive ROS, inhibits NF-κB nuclear translocation and down-regulates the TXNIP/NLRP3 axis, consequently suppressing ROS-mediated NLRP3 inflammasome activation, which may be the underlying mechanism of the protective effect of autophagy on realgar-induced liver injury. In conclusion, the results of this study suggest that autophagy alleviates realgar-induced liver injury by inhibiting ROS-mediated NLRP3 inflammasome activation. Autophagy may represent a therapeutic target in modulating realgar-induced liver injury.
    Keywords:  NLRP3 inflammasome; autophagy; liver injury; realgar
    DOI:  https://doi.org/10.3390/ijms23105697
  57. Front Aging Neurosci. 2022 ;14 897688
    Beware of Misdelivery: Multifaceted Role of Retromer Transport in Neurodegenerative Diseases.
    Shun Yoshida, Takafumi Hasegawa.
      Retromer is a highly integrated multimeric protein complex that mediates retrograde cargo sorting from endosomal compartments. In concert with its accessory proteins, the retromer drives packaged cargoes to tubular and vesicular structures, thereby transferring them to the trans-Golgi network or to the plasma membrane. In addition to the endosomal trafficking, the retromer machinery participates in mitochondrial dynamics and autophagic processes and thus contributes to cellular homeostasis. The retromer components and their associated molecules are expressed in different types of cells including neurons and glial cells, and accumulating evidence from genetic and biochemical studies suggests that retromer dysfunction is profoundly involved in the pathogenesis of neurodegenerative diseases including Alzheimer's Disease and Parkinson's disease. Moreover, targeting retromer components could alleviate the neurodegenerative process, suggesting that the retromer complex may serve as a promising therapeutic target. In this review, we will provide the latest insight into the regulatory mechanisms of retromer and discuss how its dysfunction influences the pathological process leading to neurodegeneration.
    Keywords:  Alzheimer’s Disease; Parkinson’s disease; membrane trafficking; neurodegeneration; retromer
    DOI:  https://doi.org/10.3389/fnagi.2022.897688
  58. Front Cell Dev Biol. 2022 ;10 900777
    Myocilin Gene Mutation Induced Autophagy Activation Causes Dysfunction of Trabecular Meshwork Cells.
    Xuejing Yan, Shen Wu, Qian Liu, Ying Cheng, Jingxue Zhang, Ningli Wang.
      Trabecular meshwork dysfunction is the main cause of primary open angle glaucoma (POAG) associated with elevated intraocular pressure (IOP). Mutant myocilin causes glaucoma mainly via elevating IOP. Previously we have found that accumulated Asn 450 Tyr (N450Y) mutant myocilin impairs human trabecular meshwork (TM) cells by inducing chronic endoplasmic reticulum (ER) stress response in vitro. However, it is unclear how ER stress leads to TM damage and whether N450Y myocilin mutation is associated with POAG in vivo. Here we found that N450Y mutant myocilin induces autophagy, which worsens cell viability, whereas inhibition of autophagy increases viability and decreases cell death in human TM cells. Furthermore, we construct a transgenic mouse model of N450Y myocilin mutation (Tg-MYOCN450Y) and Tg-MYOCN450Y mice exhibiting glaucoma phenotypes: IOP elevation, retinal ganglion cell loss and visual impairment. Consistent with our published in vitro studies, mutant myocilin fails to secrete into aqueous humor, causes ER stress and actives autophagy in Tg-MYOCN450Y mice, and aqueous humor dynamics are altered in Tg-MYOCN450Y mice. In summary, our studies demonstrate that activation of autophagy is correlated with pathogenesis of POAG.
    Keywords:  ER stress; POAG; autophagy; myocilin; trabecular meshwork
    DOI:  https://doi.org/10.3389/fcell.2022.900777
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