bims-auttor Biomed News
on Autophagy and mTOR
Issue of 2021‒03‒28
fifty-one papers selected by
Viktor Korolchuk, Newcastle University
  1. Fas-associated factor 1 induces the accumulation of α-synuclein through autophagic suppression in dopaminergic neurons.
  2. Inhibition of GSK-3 ameliorates the pathogenesis of Huntington's disease.
  3. mTORC1 stimulates cell growth through SAM synthesis and m6A mRNA-dependent control of protein synthesis.
  4. Demonstrating Ligandability of the LC3A and LC3B Adapter Interface.
  5. TFEB-GDF15 axis protects against obesity and insulin resistance as a lysosomal stress response.
  6. Protein phosphatase 6 dissociates the Beclin 1/Vps34 complex and inhibits autophagy.
  7. Cathepsin B and D deficiency in the mouse pancreas induces impaired autophagy and chronic pancreatitis.
  8. Autophagy-dependent survival is controlled with a unique regulatory network upon various cellular stress events.
  9. AKR1B10 negatively regulates autophagy through reducing GAPDH upon glucose starvation in colon cancer.
  10. mTORC1 promotes cell growth via m6A-dependent mRNA degradation.
  11. The Role of Mitophagy in the Regulation of Mitochondrial Energetic Status in Neurons.
  12. Evolution and insights into the structure and function of the DedA superfamily containing TMEM41B and VMP1.
  13. Plasmodium UIS3 avoids host cell-autonomous exclusion that requires GABARAPs but not LC3 and autophagy.
  14. Primary cilia and the reciprocal activation of AKT and SMAD2/3 regulate stretch-induced autophagy in trabecular meshwork cells.
  15. Pregnancy associated plasma protein-aa regulates endoplasmic reticulum-mitochondria associations.
  16. CGAS is a micronucleophagy receptor for the clearance of micronuclei.
  17. Mitochondrial network remodeling: an important feature of myogenesis and skeletal muscle regeneration.
  18. Dissolving the Complex Role Aggregation Plays in Neurodegenerative Disease.
  19. Lysosomal Stress Response (LSR): Physiological Importance and Pathological Relevance.
  20. Staufen1 in human neurodegeneration.
  21. Autophagy inhibition perturbs ERBB2 trafficking and abolishes tumorigenesis in ERBB2-driven breast cancer.
  22. Golgi maturation-dependent glycoenzyme recycling controls glycosphingolipid biosynthesis and cell growth via GOLPH3.
  23. Inhibition of chaperone‑mediated autophagy reduces tumor growth and metastasis and promotes drug sensitivity in colorectal cancer.
  24. Fatty acid oxidation and autophagy promote endoxifen resistance and counter the effect of AKT inhibition in ER-positive breast cancer cells.
  25. Autophagy Mediates Leptin-Induced Migration and ERK Activation in Breast Cancer Cells.
  26. Host-commensal interaction promotes health and lifespan in Caenorhabditis elegans through the activation of HLH-30/TFEB-mediated autophagy.
  27. Regulation of Age-Related Protein Toxicity.
  28. Is targeting autophagy mechanism in cancer a good approach? The possible double-edge sword effect.
  29. Akt-mTOR hypoactivity in bipolar disorder gives rise to cognitive impairments associated with altered neuronal structure and function.
  30. Targeting Autophagy in Neurodegenerative Diseases: from Molecular Mechanisms to Clinical Therapeutics.
  31. Global phosphoproteomics pinpoints uncharted Gcn2-mediated mechanisms of translational control.
  32. Inhibition of HMGB1 Suppresses Hepatocellular Carcinoma Progression via HIPK2-Mediated Autophagic Degradation of ZEB1.
  33. Luteolin Induces Apoptosis and Autophagy in HCT116 Colon Cancer Cells via p53-Dependent Pathway.
  34. Salvage NAD+ Biosynthetic Pathway Enzymes Moonlight as Molecular Chaperones to Protect Against Proteotoxicity.
  35. Parkinson's disease and mitophagy: an emerging role for LRRK2.
  36. Leishmania (V.) braziliensis infection promotes macrophage autophagy by a LC3B-dependent and BECLIN1-independent mechanism.
  37. VPS35 D620N knockin mice recapitulate cardinal features of Parkinson's disease.
  38. Increased LRRK2 kinase activity alters neuronal autophagy by disrupting the axonal transport of autophagosomes.
  39. DJ-1 deficiency in hepatocytes improves liver ischemia-reperfusion injury by enhancing mitophagy.
  40. Evolution of mammalian longevity: age-related increase in autophagy in bats compared to other mammals.
  41. STK3/STK4 signalling in adipocytes regulates mitophagy and energy expenditure.
  42. Modulation of the IGF1R-MTOR pathway attenuates motor neuron toxicity of human ALS SOD1G93A astrocytes.
  43. The oncogene AAMDC links PI3K-AKT-mTOR signaling with metabolic reprograming in estrogen receptor-positive breast cancer.
  44. Autophagy-lysosomal signaling responses to heat stress in tenotomy-induced rat skeletal muscle atrophy.
  45. Enhancing lifespan of budding yeast by pharmacological lowering of amino acid pools.
  46. ER-phagy responses in yeast, plants, and mammalian cells and their crosstalk with UPR and ERAD.
  47. Arbutin ameliorates glucocorticoid-induced osteoporosis through activating autophagy in osteoblasts.
  48. Autophagy in tumour immunity and therapy.
  49. PARsylated transcription factor EB (TFEB) regulates the expression of a subset of Wnt target genes by forming a complex with β-catenin-TCF/LEF1.
  50. The upregulation of Ulk1-dependent autophagy does not require the p53 activity in mouse embryonic stem cells.
  51. ATG16L1 functions in cell homeostasis beyond autophagy.
  1. FASEB J. 2021 Apr;35(4): e21363
    Fas-associated factor 1 induces the accumulation of α-synuclein through autophagic suppression in dopaminergic neurons.
    Bok-Seok Kim, Taeik Jang, Sung-Eun Yoo, Jae Moon Lee, Eunhee Kim.
      Impairment of protein clearance mechanisms leads to α-synuclein accumulation in dopaminergic neurons, contributing to the pathogenesis of Parkinson's disease (PD). Based on the finding that Fas-associated factor 1 (FAF1), a positive modulator of PD, colocalizes with α-synuclein in PD patient brains, we investigated the existence of pathological interplay between FAF1 and α-synuclein. Monomeric and high-molecular-weight forms of α-synuclein were increased in FAF1-overexpressing SH-SY5Y cells. In particular, α-synuclein turnover was accelerated by genetic depletion of FAF1 in SH-SY5Y cells. Therefore, we questioned whether FAF1 is involved in the α-synuclein clearance process. Autophagy inhibitors, but not proteasome inhibitors, restored concurrent attenuation of α-synuclein expression by FAF1 depletion in SH-SY5Y cells. Moreover, we found alterations in autophagy markers in SH-SY5Y cells caused by FAF1 overexpression, indicating that FAF1 disturbed α-synuclein clearance through the autophagy-lysosome pathway. Indeed, FAF1 activated the mammalian target of rapamycin (mTOR) pathway, subsequently suppressing autophagosome formation. Consistently, α-synuclein-mediated mitochondrial dysfunction was observed in FAF1-overexpressing SH-SY5Y cells. Furthermore, FAF1 overexpression using stereotaxic injection of adeno-associated virus led to α-synuclein accumulation and autophagy dysregulation in the PD model mice. Taken together, our results reveal a novel role for FAF1: that of a negative regulator of autophagic α-synuclein clearance.
    Keywords:  Parkinson's disease; macroautophagy; striatum; substantia nigra
    DOI:  https://doi.org/10.1096/fj.202001371RR
  2. Neurobiol Dis. 2021 Mar 19. pii: S0969-9961(21)00085-1. [Epub ahead of print] 105336
    Inhibition of GSK-3 ameliorates the pathogenesis of Huntington's disease.
    Ido Rippin, Katherina Bonder, Shirley Ben Joseph, Amar Sarsor, Lilach Vaks, Hagit Eldar-Finkelman.
      In Huntington's disease (HD), the mutant huntingtin (mHtt) accumulates as toxic aggregates in the striatum tissue, with deleterious effects on motor-coordination and cognitive functions. Reducing the levels of mHtt is therefore a promising therapeutic strategy. We have previously reported that GSK-3 is a negative regulator of the autophagy/lysosome pathway, which is responsible for intracellular degradation, and is critically important for maintaining neuronal vitality. Thus, we hypothesized that inhibition of GSK-3 may trigger mHtt clearance thereby reducing mHtt cytotoxicity and improving HD symptoms. Here, we demonstrate that depletion or suppression of autophagy results in a massive accumulation of mHtt aggregates. Accordingly, mHtt aggregates were localized in lysosomes, but, mostly mislocalized from lysosomes in the absence of functional autophagy. Overexpression of GSK-3, particularly the α isozyme, increased the number of mHtt aggregates, while silencing GSK-3α/β, or treatment with a selective GSK-3 inhibitor, L807mts, previously described by us, reduced the amounts of mHtt aggregates. This effect was mediated by increased autophagic and lysosomal activity. Treating R6/2 mouse model of HD with L807mts, reduced striatal mHtt aggregates and elevated autophagic and lysosomal markers. The L807mts treatment also reduced hyperglycemia and improved motor-coordination functions in these mice. In addition, L807mts restored the expression levels of Sirt1, a critical neuroprotective factor in the HD striatum, along with its targets BDNF, DRPP-32, and active Akt, all provide neurotrophic/pro-survival support and typically decline in the HD brain. Our results provide strong evidence for a role for GSK-3 in the regulation of mHtt dynamics, and demonstrate the benefits of GSK-3 inhibition in reducing mHtt toxicity, providing neuroprotective support, and improving HD symptoms.
    Keywords:  Autophagy; GSK-3; GSK-3 inhibitor; Huntington's diseases; L807mts; Lysosome; Mutant huntingtin; R6/2 mice; Sirt1
    DOI:  https://doi.org/10.1016/j.nbd.2021.105336
  3. Mol Cell. 2021 Mar 17. pii: S1097-2765(21)00177-5. [Epub ahead of print]
    mTORC1 stimulates cell growth through SAM synthesis and m6A mRNA-dependent control of protein synthesis.
    Elodie Villa, Umakant Sahu, Brendan P O'Hara, Eunus S Ali, Kathryn A Helmin, John M Asara, Peng Gao, Benjamin D Singer, Issam Ben-Sahra.
      The mechanistic target of rapamycin complex 1 (mTORC1) regulates metabolism and cell growth in response to nutrient, growth, and oncogenic signals. We found that mTORC1 stimulates the synthesis of the major methyl donor, S-adenosylmethionine (SAM), through the control of methionine adenosyltransferase 2 alpha (MAT2A) expression. The transcription factor c-MYC, downstream of mTORC1, directly binds to intron 1 of MAT2A and promotes its expression. Furthermore, mTORC1 increases the protein abundance of Wilms' tumor 1-associating protein (WTAP), the positive regulatory subunit of the human N6-methyladenosine (m6A) RNA methyltransferase complex. Through the control of MAT2A and WTAP levels, mTORC1 signaling stimulates m6A RNA modification to promote protein synthesis and cell growth. A decline in intracellular SAM levels upon MAT2A inhibition decreases m6A RNA modification, protein synthesis rate, and tumor growth. Thus, mTORC1 adjusts m6A RNA modification through the control of SAM and WTAP levels to prime the translation machinery for anabolic cell growth.
    Keywords:  Cell growth; MAT2A; Methionine cycle; N(6)-methyladenosine; Protein Synthesis; RNA metabolism; S-adenosylmethionine; WTAP; mTOR; mTORC1
    DOI:  https://doi.org/10.1016/j.molcel.2021.03.009
  4. J Med Chem. 2021 Mar 26.
    Demonstrating Ligandability of the LC3A and LC3B Adapter Interface.
    Markus Hartmann, Jessica Huber, Jan S Kramer, Jan Heering, Larissa Pietsch, Holger Stark, Dalibor Odadzic, Iris Bischoff, Robert Fürst, Martin Schröder, Masato Akutsu, Apirat Chaikuad, Volker Dötsch, Stefan Knapp, Ricardo M Biondi, Vladimir V Rogov, Ewgenij Proschak.
      Autophagy is the common name for a number of lysosome-based degradation pathways of cytosolic cargos. The key components of autophagy are members of Atg8 family proteins involved in almost all steps of the process, from autophagosome formation to their selective fusion with lysosomes. In this study, we show that the homologous members of the human Atg8 family proteins, LC3A and LC3B, are druggable by a small molecule inhibitor novobiocin. Structure-activity relationship (SAR) studies of the 4-hydroxy coumarin core scaffold were performed, supported by a crystal structure of the LC3A dihydronovobiocin complex. The study reports the first nonpeptide inhibitors for these protein interaction targets and will lay the foundation for the development of more potent chemical probes for the Atg8 protein family which may also find applications for the development of autophagy-mediated degraders (AUTACs).
    DOI:  https://doi.org/10.1021/acs.jmedchem.0c01564
  5. Nat Metab. 2021 Mar;3(3): 410-427
    TFEB-GDF15 axis protects against obesity and insulin resistance as a lysosomal stress response.
    Jinyoung Kim, Seong Hun Kim, Hyereen Kang, Soyeon Lee, Shi-Young Park, Yoonil Cho, Yu-Mi Lim, Ji Woong Ahn, Young-Hwan Kim, Seungsoo Chung, Cheol Soo Choi, Yeon Jin Jang, Hye Soon Park, Yoonseok Heo, Kook Hwan Kim, Myung-Shik Lee.
      TFEB, a key regulator of lysosomal biogenesis and autophagy, is induced not only by nutritional deficiency but also by organelle stress. Here, we find that Tfeb and its downstream genes are upregulated together with lipofuscin accumulation in adipose tissue macrophages (ATMs) of obese mice or humans, suggestive of obesity-associated lysosomal dysfunction/stress in ATMs. Macrophage-specific TFEB-overexpressing mice display complete abrogation of diet-induced obesity, adipose tissue inflammation and insulin resistance, which is independent of autophagy, but dependent on TFEB-induced GDF15 expression. Palmitic acid induces Gdf15 expression through lysosomal Ca2+-mediated TFEB nuclear translocation in response to lysosomal stress. In contrast, mice fed a high-fat diet with macrophage-specific Tfeb deletion show aggravated adipose tissue inflammation and insulin resistance, accompanied by reduced GDF15 level. Finally, we observe activation of TFEB-GDF15 in ATMs of obese humans as a consequence of lysosomal stress. These findings highlight the importance of the TFEB-GDF15 axis as a lysosomal stress response in obesity or metabolic syndrome and as a promising therapeutic target for treatment of these conditions.
    DOI:  https://doi.org/10.1038/s42255-021-00368-w
  6. Biochem Biophys Res Commun. 2021 Mar 19. pii: S0006-291X(21)00364-8. [Epub ahead of print]552 191-195
    Protein phosphatase 6 dissociates the Beclin 1/Vps34 complex and inhibits autophagy.
    Nobuyuki Fujiwara, Shusaku Shibutani, Takashi Ohama, Koichi Sato.
      Autophagy is an evolutionarily conserved intracellular degradation system and is regulated by various signaling pathways including the Beclin 1/Vacuolar protein sorting 34 (Vps34) complex. Protein phosphatase 6 (PP6) is an essential serine/threonine phosphatase that regulates various biological processes. Recently, we found that PP6 protein is degraded by p62-dependent selective autophagy. In this study, we show that PP6 conversely inhibits autophagy. PP6 associate with the C-terminal region of Beclin 1, which is close to the binding region of Vps34. The protein levels of PP6 affect Beclin 1/Vps34 complex formation and phosphatase activity of PP6 is not involved in this. We also show that chemically induced PP6/Beclin 1 association leads to Vps34 dissociation from Beclin 1. Overall, our data reveal a novel regulatory mechanism for autophagy by PP6.
    Keywords:  Autophagy; Beclin 1; Protein-protein interaction; Serine/threonine protein phosphatase 6
    DOI:  https://doi.org/10.1016/j.bbrc.2021.02.136
  7. Sci Rep. 2021 Mar 23. 11(1): 6596
    Cathepsin B and D deficiency in the mouse pancreas induces impaired autophagy and chronic pancreatitis.
    Hideaki Iwama, Sally Mehanna, Mai Imasaka, Shinsuke Hashidume, Hiroshi Nishiura, Ken-Ichi Yamamura, Chigure Suzuki, Yasuo Uchiyama, Etsuro Hatano, Masaki Ohmuraya.
      The major lysosomal proteases, Cathepsin B (CTSB), Cathepsin D (CTSD) and Cathepsin L (CTSL), are implicated in autophagic activity. To investigate the role of each cathepsin in the exocrine pancreas, we generated mice in which the pancreas was specifically deficient in Ctsb, Ctsd and Ctsl. Each of these gene knockout (KO) and Ctsb;Ctsl and Ctsd;Ctsl double-knockout (DKO) mice were almost normal. However, we found cytoplasmic degeneration in the pancreatic acinar cells of Ctsb;Ctsd DKO mice, similar to autophagy related 5 (Atg5) KO mice. LC3 and p62 (autophagy markers) showed remarkable accumulation and the numbers of autophagosomes and autolysosomes were increased in the pancreatic acinar cells of Ctsb;Ctsd DKO mice. Moreover, these Ctsb;Ctsd DKO mice also developed chronic pancreatitis (CP). Thus, we conclude that both Ctsb and Ctsd deficiency caused impaired autophagy in the pancreatic acinar cells, and induced CP in mice.
    DOI:  https://doi.org/10.1038/s41598-021-85898-9
  8. Cell Death Dis. 2021 Mar 23. 12(4): 309
    Autophagy-dependent survival is controlled with a unique regulatory network upon various cellular stress events.
    Orsolya Kapuy, Marianna Holczer, Margita Márton, Tamás Korcsmáros.
      Although autophagy is a type of programmed cell death, it is also essential for cell survival upon tolerable level of various stress events. For the cell to respond adequately to an external and/or internal stimulus induced by cellular stress, autophagy must be controlled in a highly regulated manner. By using systems biology techniques, here we explore the dynamical features of autophagy induction. We propose that the switch-like characteristic of autophagy induction is achieved by a control network, containing essential feedback loops of four components, so-called autophagy inducer, autophagy controller, mTORC1 and autophagy executor, respectively. We show how an autophagy inducer is capable to turn on autophagy in a cellular stress-specific way. The autophagy controller acts as a molecular switch and not only promotes autophagy but also blocks the permanent hyperactivation of the process via downregulating the autophagy inducer. In this theoretical analysis, we explore in detail the properties of all four proposed controlling elements and their connections. Here we also prove that the kinetic features of this control network can be considered accurate in various stress processes (such as starvation, endoplasmic reticulum stress and oxidative stress), even if the exact components may be different. The robust response of the resulting control network is essential during cellular stress.
    DOI:  https://doi.org/10.1038/s41419-021-03599-7
  9. J Cell Sci. 2021 Mar 23. pii: jcs.255273. [Epub ahead of print]
    AKR1B10 negatively regulates autophagy through reducing GAPDH upon glucose starvation in colon cancer.
    Wanyun Li, Cong Liu, Zilan Huang, Lei Shi, Chuanqi Zhong, Wenwen Zhou, Peipei Meng, Zhenyu Li, Shengyu Wang, Fanghong Luo, Jianghua Yan, Ting Wu.
      Autophagy is considered as an important switch for cell transformation from normal to malignant during colorectal cancer development. Consistent with other reports, we found the membrane receptor Neuropilin1 (NRP1) is greatly upregulated in colon cancer cells that underwent autophagy upon glucose deprivation. However, the mechanism underlying NRP1 regulation of autophagy is unknown. We found that knockdown of the NRP1 inhibits autophagy and largely upregulates the expression of Aldo-Keto Reductase family 1 B10 (AKR1B10). Moreover, we demonstrated that AKR1B10 interacts with and inhibits the nuclear import of Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and then subsequently represses autophagy. Interestingly, we also found a NADPH-dependent reduction reaction could be induced when AKR1B10 interacts with GAPDH and the reductase activity of AKR1B10 is important for its repressing autophagy. Together, our findings unravel a novel mechanism of NRP1 in regulating autophagy through AKR1B10.
    Keywords:  Aldo-Keto Reductase family 1 B10 (AKR1B10); Glucose starvation; Glyceraldehyde-3-phosphate dehydrogenase (GAPDH); Neuropilin1 (NRP1); Nuclear translocation; Reductase activity
    DOI:  https://doi.org/10.1242/jcs.255273
  10. Mol Cell. 2021 Mar 19. pii: S1097-2765(21)00178-7. [Epub ahead of print]
    mTORC1 promotes cell growth via m6A-dependent mRNA degradation.
    Sungyun Cho, Gina Lee, Brian F Pickering, Cholsoon Jang, Jin H Park, Long He, Lavina Mathur, Seung-Soo Kim, Sunhee Jung, Hong-Wen Tang, Sebastien Monette, Joshua D Rabinowitz, Norbert Perrimon, Samie R Jaffrey, John Blenis.
      Dysregulated mTORC1 signaling alters a wide range of cellular processes, contributing to metabolic disorders and cancer. Defining the molecular details of downstream effectors is thus critical for uncovering selective therapeutic targets. We report that mTORC1 and its downstream kinase S6K enhance eIF4A/4B-mediated translation of Wilms' tumor 1-associated protein (WTAP), an adaptor for the N6-methyladenosine (m6A) RNA methyltransferase complex. This regulation is mediated by 5' UTR of WTAP mRNA that is targeted by eIF4A/4B. Single-nucleotide-resolution m6A mapping revealed that MAX dimerization protein 2 (MXD2) mRNA contains m6A, and increased m6A modification enhances its degradation. WTAP induces cMyc-MAX association by suppressing MXD2 expression, which promotes cMyc transcriptional activity and proliferation of mTORC1-activated cancer cells. These results elucidate a mechanism whereby mTORC1 stimulates oncogenic signaling via m6A RNA modification and illuminates the WTAP-MXD2-cMyc axis as a potential therapeutic target for mTORC1-driven cancers.
    Keywords:  MXD2; Protein translation; S6K1; WTAP; YTHDF readers; cMyc; eIF4A; m(6)A mRNA modification; mRNA stability; mTORC1
    DOI:  https://doi.org/10.1016/j.molcel.2021.03.010
  11. Autophagy. 2021 Mar 23.
    The Role of Mitophagy in the Regulation of Mitochondrial Energetic Status in Neurons.
    Sinsuk Han, Mingyang Zhang, Yu Young Jeong, David Margolis, Qian Cai.
      Mitochondria are the main cellular energy powerhouses and supply most of the energy in the form of ATP to fuel essential neuronal functions through oxidative phosphorylation (OXPHOS). In Alzheimer disease (AD), metabolic and mitochondrial disruptions are an early feature preceding any histopathological and clinical manifestations. Mitochondrial malfunction is also linked to synaptic defects in early AD. Mitophagy serves as a key cellular quality control mechanism involving sequestration of damaged mitochondria within autophagosomes and their subsequent degradation in lysosomes. However, it remains largely unknown whether mitophagy is involved in the regulation of energy metabolism in neurons, and if so, whether metabolic deficiency in AD is attributed to mitophagy dysfunction. Here we reveal that mitophagy is broadly activated in metabolically enhanced neurons upon OXPHOS stimulation, which sustains high energetic activity by increasing mitochondrial turnover and hence facilitating mitochondrial maintenance. Unexpectedly, in AD-related mutant HsAPP Tg mouse brains, early stimulation of OXPHOS activity fails to correct energy deficits but exacerbates synapse loss as a consequence of mitophagy failure. Excitingly, lysosomal enhancement in AD neurons restores impaired metabolic function by promoting elimination of damaged mitochondria, protecting against synaptic damage in AD mouse brains. Taken together, we propose a new mechanism by which mitophagy controls bioenergetic status in neurons, furthering our understanding of the direct impact of mitophagy defects on AD-linked metabolic deficits and shedding light on the development of novel therapeutic strategies to treat AD by the early stimulation of mitochondrial metabolism combined with elevation of lysosomal proteolytic activity.
    Keywords:  Alzheimer; bioenergetics; energy metabolism; lysosomal proteolysis; metabolic deficiency; mitochondrial stress; mitophagosome; neuronal mitophagy; retrograde transport; synapse loss
    DOI:  https://doi.org/10.1080/15548627.2021.1907167
  12. J Cell Sci. 2021 Mar 26. pii: jcs.255877. [Epub ahead of print]
    Evolution and insights into the structure and function of the DedA superfamily containing TMEM41B and VMP1.
    Fumiya Okawa, Yutaro Hama, Sidi Zhang, Hideaki Morishita, Hayashi Yamamoto, Tim P Levine, Noboru Mizushima.
      TMEM41B and VMP1 are endoplasmic reticulum (ER)-localizing multi-spanning membrane proteins required for ER-related cellular processes such as autophagosome formation, lipid droplet homeostasis, and lipoprotein secretion in eukaryotes. Both proteins have a VTT domain, which is similar to the DedA domain found in bacterial DedA family proteins. However, the molecular function and structure of the DedA and VTT domains (collectively referred to as DedA domains) and the evolutionary relationships among the DedA domain-containing proteins are largely unknown. Here, we conduct remote homology search and identify a new clade consisting mainly of bacterial PF06695 proteins of unknown function. Phylogenetic analysis reveals that the TMEM41, VMP1, DedA, and PF06695 families form a superfamily with a common origin, which we term the DedA superfamily. Coevolution-based structural prediction suggests that the DedA domain contains two reentrant loops facing each other in the membrane. This topology is biochemically verified by the substituted cysteine accessibility method. The predicted structure is topologically similar to that of the substrate-binding region of Na+-coupled glutamate transporter solute carrier 1. A potential ion-coupled transport function of the DedA superfamily proteins is discussed.
    Keywords:  Autophagy; DedA; Reentrant loop; TMEM41B; VMP1
    DOI:  https://doi.org/10.1242/jcs.255877
  13. Parasitol Int. 2021 Mar 23. pii: S1383-5769(21)00054-4. [Epub ahead of print] 102335
    Plasmodium UIS3 avoids host cell-autonomous exclusion that requires GABARAPs but not LC3 and autophagy.
    Ariel Pradipta, Hironori Bando, Ji Su Ma, Shun Tanaka, Miwa Sasai, Masahiro Yamamoto.
      Sporozoites of the etiological agent of malaria, Plasmodium, form parasitophorous vacuoles (PVs) in hepatocytes. The PV membranes (PVM) are coated with a well-known host autophagy marker LC3 and parasite-derived protein called Upregulated in infective sporozoites 3 (UIS3), which has been shown to interact with LC3 and inhibit LC3-mediated autophagic disruption at the PV. Although uis3(-) sporozoites cannot proliferate in wild-type cells, they can replicate efficiently in cells defective in autophagy due to the lack of Atg proteins such as Atg3, Atg5 and Atg7, since these Atg proteins are essential for processing of LC3. However, it remains to be seen whether other Atg proteins participate in the restriction of uis3(-) parasite growth. Here we show that, despite essential roles of Atg9 and Atg14 in autophagy, both proteins are dispensable for the restriction of uis3(-) parasite growth. Moreover, we found that cells lacking LC3 proteins are also able to restrict uis3(-) parasite growth. In sharp contrast, GABARAPs, another subfamily of mammalian Atg8, participated in suppression of uis3(-) parasite growth. Taken together, contrary to a previous model in which UIS3 avoids host LC3- and autophagy-dependent parasite elimination program, our data demonstrate a role of GABARAPs for suppression of uis3(-) parasite growth in a manner independent on autophagy.
    Keywords:  Atg proteins; Autophagy; GABARAPs; LC3s; Plasmodium berghei; Upregulated in infective sporozoites 3 (UIS3)
    DOI:  https://doi.org/10.1016/j.parint.2021.102335
  14. Proc Natl Acad Sci U S A. 2021 Mar 30. pii: e2021942118. [Epub ahead of print]118(13):
    Primary cilia and the reciprocal activation of AKT and SMAD2/3 regulate stretch-induced autophagy in trabecular meshwork cells.
    Myoung Sup Shim, April Nettesheim, Angela Dixon, Paloma B Liton.
      Activation of autophagy is one of the responses elicited by high intraocular pressure (IOP) and mechanical stretch in trabecular meshwork (TM) cells. However, the mechanosensor and the molecular mechanisms by which autophagy is induced by mechanical stretch in these or other cell types is largely unknown. Here, we have investigated the mechanosensor and downstream signaling pathway that regulate cyclic mechanical stretch (CMS)-induced autophagy in TM cells. We report that primary cilia act as a mechanosensor for CMS-induced autophagy and identified a cross-regulatory talk between AKT1 and noncanonical SMAD2/3 signaling as critical components of primary cilia-mediated activation of autophagy by mechanical stretch. Furthermore, we demonstrated the physiological significance of our findings in ex vivo perfused eyes. Removal of primary cilia disrupted the homeostatic IOP compensatory response and prevented the increase in LC3-II protein levels in response to elevated pressure challenge, strongly supporting a role of primary cilia-mediated autophagy in regulating IOP homeostasis.
    Keywords:  autophagy; glaucoma; primary cilia; stretching; trabecular meshwork
    DOI:  https://doi.org/10.1073/pnas.2021942118
  15. Elife. 2021 Mar 24. pii: e59687. [Epub ahead of print]10
    Pregnancy associated plasma protein-aa regulates endoplasmic reticulum-mitochondria associations.
    Mroj Alassaf, Mary C Halloran.
      Endoplasmic reticulum (ER) and mitochondria form close physical associations to facilitate calcium transfer, thereby regulating mitochondrial function. Neurons with high metabolic demands, such as sensory hair cells, are especially dependent on precisely regulated ER-mitochondria associations. We previously showed that the secreted metalloprotease Pregnancy associated plasma protein-aa (Pappaa) regulates mitochondrial function in zebrafish lateral line hair cells (Alassaf et al., 2019). Here, we show that pappaa mutant hair cells exhibit excessive and abnormally close ER-mitochondria associations, suggesting increased ER-mitochondria calcium transfer. pappaa mutant hair cells are more vulnerable to pharmacological induction of ER-calcium transfer. Additionally, pappaa mutant hair cells display ER stress and dysfunctional downstream processes of the ER-mitochondria axis including altered mitochondrial morphology and reduced autophagy. We further show that Pappaa influences ER-calcium transfer and autophagy via its ability to stimulate insulin-like growth factor-1 bioavailability. Together our results identify Pappaa as a novel regulator of the ER-mitochondria axis.
    Keywords:  cell biology; neuroscience; zebrafish
    DOI:  https://doi.org/10.7554/eLife.59687
  16. Autophagy. 2021 Mar 22. 1-17
    CGAS is a micronucleophagy receptor for the clearance of micronuclei.
    Mengmeng Zhao, Fei Wang, Juehui Wu, Yuanna Cheng, Yajuan Cao, Xiangyang Wu, Mingtong Ma, Fen Tang, Zhi Liu, Haipeng Liu, Baoxue Ge.
      Micronuclei are constantly considered as a marker of genome instability and very recently found to be a trigger of innate immune responses. An increased frequency of micronuclei is associated with many diseases, but the mechanism underlying the regulation of micronuclei homeostasis remains largely unknown. Here, we report that CGAS (cyclic GMP-AMP synthase), a known regulator of DNA sensing and DNA repair, reduces the abundance of micronuclei under genotoxic stress in an autophagy-dependent manner. CGAS accumulates in the autophagic machinery and directly interacts with MAP1LC3B/LC3B in a manner dependent upon its MAP1LC3-interacting region (LIR). Importantly, the interaction is essential for MAP1LC3 recruitment to micronuclei and subsequent clearance of micronuclei via autophagy (micronucleophagy) in response to genotoxic stress. Moreover, in contrast to its DNA sensing function to activate micronuclei-driven inflammation, CGAS-mediated micronucleophagy blunts the production of cyclic GMP-AMP (cGAMP) induced by genotoxic stress. We therefore conclude that CGAS is a receptor for the selective autophagic clearance of micronuclei and uncovered an unprecedented role of CGAS in micronuclei homeostasis to dampen innate immune surveillance.
    Keywords:  CGAS; MAP1LC3B/LC3B; autophagy receptor; micronuclei; micronucleophagy
    DOI:  https://doi.org/10.1080/15548627.2021.1899440
  17. Cell Mol Life Sci. 2021 Mar 22.
    Mitochondrial network remodeling: an important feature of myogenesis and skeletal muscle regeneration.
    Fasih Ahmad Rahman, Joe Quadrilatero.
      The remodeling of the mitochondrial network is a critical process in maintaining cellular homeostasis and is intimately related to mitochondrial function. The interplay between the formation of new mitochondria (biogenesis) and the removal of damaged mitochondria (mitophagy) provide a means for the repopulation of the mitochondrial network. Additionally, mitochondrial fission and fusion serve as a bridge between biogenesis and mitophagy. In recent years, the importance of these processes has been characterised in multiple tissue- and cell-types, and under various conditions. In skeletal muscle, the robust remodeling of the mitochondrial network is observed, particularly after injury where large portions of the tissue/cell structures are damaged. The significance of mitochondrial remodeling in regulating skeletal muscle regeneration has been widely studied, with alterations in mitochondrial remodeling processes leading to incomplete regeneration and impaired skeletal muscle function. Needless to say, important questions related to mitochondrial remodeling and skeletal muscle regeneration still remain unanswered and require further investigation. Therefore, this review will discuss the known molecular mechanisms of mitochondrial network remodeling, as well as integrate these mechanisms and discuss their relevance in myogenesis and regenerating skeletal muscle.
    Keywords:  Biogenesis; Fission; Fusion; Mitochondria; Mitophagy; Regeneration; Skeletal muscle; Skeletal muscle stem cells
    DOI:  https://doi.org/10.1007/s00018-021-03807-9
  18. Mov Disord. 2021 Mar 23.
    Dissolving the Complex Role Aggregation Plays in Neurodegenerative Disease.
    Katherine R Croce, Ai Yamamoto.
      Prominent neuropathological hallmarks of many adult-onset neurodegenerative diseases include the deposition and accumulation of misfolded proteins or conformers; however, their role in pathogenesis has remained unclear. This is in part due to the deceptive simplicity of the question and our limited understanding of how protein homeostasis is maintained in the compartmentalized cells of the central nervous system, especially in the context of the adult brain. Building on studies from simple cell-based systems and invertebrate animals, we recently identified a protein central to the specific and selective turnover of aggregated proteins in the adult brain, the autophagy-linked FYVE protein (Alfy)/Wdfy3. Depletion of Alfy levels in a mouse model of Huntington's disease showed that it accelerated the accumulation of the aggregated mutant huntingtin protein, as well as the onset of behavioral deficits. Although the motor dysfunction was accelerated in the model, this was in the absence of increasing overt cell loss, implicating protein aggregates as a modifier of circuit dysfunction rather than driving degeneration per se. We discuss these findings in the context of what is known about protein accumulation and how we can use proteins such as Alfy to determine if protein accumulation is a shared pathogenic event across different adult-onset diseases. © 2021 International Parkinson and Movement Disorder Society.
    Keywords:  Huntington's disease; aggregation; autophagy; mouse models; neurodegeneration
    DOI:  https://doi.org/10.1002/mds.28522
  19. J Neuroimmune Pharmacol. 2021 Mar 22.
    Lysosomal Stress Response (LSR): Physiological Importance and Pathological Relevance.
    Koffi L Lakpa, Nabab Khan, Zahra Afghah, Xuesong Chen, Jonathan D Geiger.
      Extensive work has characterized endoplasmic reticulum (ER) and mitochondrial stress responses. In contrast, very little has been published about stress responses in lysosomes; subcellular acidic organelles that are physiologically important and are of pathological relevance. The greater lysosomal system is dynamic and is comprised of endosomes, lysosomes, multivesicular bodies, autophagosomes, and autophagolysosomes. They are important regulators of cellular physiology, they represent about 5% of the total cellular volume, they are heterogeneous in their sizes and distribution patterns, they are electron dense, and their subcellular positioning within cells varies in response to stimuli, insults and pH. These organelles are also integral to the pathogenesis of lysosomal storage diseases and it is increasingly recognized that lysosomes play important roles in the pathogenesis of such diverse conditions as neurodegenerative disorders and cancer. The purpose of this review is to focus attention on lysosomal stress responses (LSR), compare LSR with better characterized stress responses in ER and mitochondria, and form a framework for future characterizations of LSR. We synthesized data into the concept of LSR and present it here such that the definition of LSR can be modified as new knowledge is added and specific therapeutics are developed.
    Keywords:  Endoplasmic reticulum stress; Endosomes; Inter-organellar signaling; Lysosomes; Mitochondrial stress
    DOI:  https://doi.org/10.1007/s11481-021-09990-7
  20. Ann Neurol. 2021 Mar 21.
    Staufen1 in human neurodegeneration.
    Sharan Paul, Warunee Dansithong, Karla P Figueroa, Mandi Gandelman, Daniel R Scoles, Stefan M Pulst.
      OBJECTIVE: Mutant expansions of DNA CAG ≥32 repeats in ATXN2 can be a rare cause of Parkinson disease and Amyotrophic lateral sclerosis (ALS). We recently reported that the stress granule (SG) protein Staufen1 (STAU1) was overabundant in neurodegenerative disorder SCA2 patient cells, animal models, and ALS-TDP-43 fibroblasts and provided a link between SG formation and autophagy. We aimed to test if STAU1 overabundance has role in the pathogenesis of other neurodegenerative diseases.METHODS: With multiple neurodegenerative patient-derived cell models, animal models, and human postmortem ALS tissue, we evaluate STAU1 function using biochemical and immunohistological analyses.
    RESULTS: We demonstrate STAU1 overabundance and increased total and phosphorylated mammalian target of rapamycin (mTOR) in fibroblast cells from ALS patients with mutations in TDP-43, dementia patients with PSEN1 mutations, a parkinsonism patient with MAPT mutation, Huntington disease (HD) mutations, and SCA2 mutations. Increased STAU1 levels and mTOR activity were seen in human ALS spinal cord tissues as well as in animal models. Changes in STAU1 and mTOR protein levels were post-transcriptional. Exogenous expression of STAU1 in wildtype cells was sufficient to activate mTOR and downstream targets and form SGs. Targeting STAU1 by RNAi normalized mTOR, suggesting a potential role for therapy in diseases associated with STAU1 overabundance.
    INTERPRETATION: STAU1 overabundance in neurodegeneration is a common phenomenon associated with hyperactive mTOR. Targeting STAU1 with ASOs or miRNA viral vectors may represent a novel, efficacious therapy for neurodegenerative diseases characterized by overabundant STAU1. This article is protected by copyright. All rights reserved.
    DOI:  https://doi.org/10.1002/ana.26069
  21. Autophagy. 2021 Mar 23.
    Autophagy inhibition perturbs ERBB2 trafficking and abolishes tumorigenesis in ERBB2-driven breast cancer.
    Mingang Hao, Syn Kok Yeo, Jun-Lin Guan.
      Macroautophagy/autophagy modulation is increasingly recognized as a potential strategy for cancer therapy. Using a recently developed Rb1cc1 mutant knockin mice model, we have taken a rigorous genetic approach to assess the role of both its autophagy and non-canonical functions in an ERBB2-driven BrCA model. We found that autophagy abrogation virtually abolishes mammary tumorigenesis in the ERBB2-driven model, exhibiting stronger inhibitory effects than in our previous studies using PyMT and brca1-null mouse models. Mechanistically, autophagy inhibition perturbs ERBB2 intracellular trafficking and triggers its release via small extracellular vesicles. Our results demonstrate a new mechanism for autophagy to promote tumorigenesis in ERBB2-driven BrCA and could supplement current strategies for anti-ERBB2 therapy.
    Keywords:  Autophagy; ERBB2-positive breast cancer; FIP200/RB1CC1; small extracellular vesicles
    DOI:  https://doi.org/10.1080/15548627.2021.1907168
  22. EMBO J. 2021 Mar 22. e107238
    Golgi maturation-dependent glycoenzyme recycling controls glycosphingolipid biosynthesis and cell growth via GOLPH3.
    Riccardo Rizzo, Domenico Russo, Kazuo Kurokawa, Pranoy Sahu, Bernadette Lombardi, Domenico Supino, Mikhail A Zhukovsky, Anthony Vocat, Prathyush Pothukuchi, Vidya Kunnathully, Laura Capolupo, Gaelle Boncompain, Carlo Vitagliano, Federica Zito Marino, Gabriella Aquino, Daniela Montariello, Petra Henklein, Luigi Mandrich, Gerardo Botti, Henrik Clausen, Ulla Mandel, Toshiyuki Yamaji, Kentaro Hanada, Alfredo Budillon, Franck Perez, Seetharaman Parashuraman, Yusuf A Hannun, Akihiko Nakano, Daniela Corda, Giovanni D'Angelo, Alberto Luini.
      Glycosphingolipids are important components of the plasma membrane where they modulate the activities of membrane proteins including signalling receptors. Glycosphingolipid synthesis relies on competing reactions catalysed by Golgi-resident enzymes during the passage of substrates through the Golgi cisternae. The glycosphingolipid metabolic output is determined by the position and levels of the enzymes within the Golgi stack, but the mechanisms that coordinate the intra-Golgi localisation of the enzymes are poorly understood. Here, we show that a group of sequentially-acting enzymes operating at the branchpoint among glycosphingolipid synthetic pathways binds the Golgi-localised oncoprotein GOLPH3. GOLPH3 sorts these enzymes into vesicles for intra-Golgi retro-transport, acting as a component of the cisternal maturation mechanism. Through these effects, GOLPH3 controls the sub-Golgi localisation and the lysosomal degradation rate of specific enzymes. Increased GOLPH3 levels, as those observed in tumours, alter glycosphingolipid synthesis and plasma membrane composition thereby promoting mitogenic signalling and cell proliferation. These data have medical implications as they outline a novel oncogenic mechanism of action for GOLPH3 based on glycosphingolipid metabolism.
    Keywords:  GOLPH3; Golgi; Trafficking; cisternal maturation; mTOR
    DOI:  https://doi.org/10.15252/embj.2020107238
  23. Mol Med Rep. 2021 May;pii: 360. [Epub ahead of print]23(5):
    Inhibition of chaperone‑mediated autophagy reduces tumor growth and metastasis and promotes drug sensitivity in colorectal cancer.
    Ying Xuan, Shuang Zhao, Xingjun Xiao, Liwei Xiang, Hua-Chuan Zheng.
      Chaperone‑mediated autophagy (CMA) is a selective type of autophagy whereby a specific subset of intracellular proteins is targeted to the lysosome for degradation. The present study investigated the mechanisms underlying the response and resistance to 5‑fluorouracil (5‑FU) in colorectal cancer (CRC) cell lines. In engineered 5‑FU‑resistant CRC cell lines, a significant elevation of lysosome‑associated membrane protein 2A (LAMP2A), which is the key molecule in the CMA pathway, was identified. High expression of LAMP2A was found to be responsible for 5‑FU resistance and to enhance PLD2 expression through the activation of NF‑κB pathway. Accordingly, loss or gain of function of LAMP2A in 5‑FU‑resistant CRC cells rendered them sensitive or resistant to 5‑FU, respectively. Taken together, the results of the present study suggested that chemoresistance in patients with CRC may be mediated by enhancing CMA. Thus, CMA is a promising predictor of chemosensitivity to 5‑FU treatment and anti‑CMA therapy may be a novel therapeutic option for patients with CRC.
    DOI:  https://doi.org/10.3892/mmr.2021.11999
  24. J Mol Cell Biol. 2021 Mar 23. pii: mjab018. [Epub ahead of print]
    Fatty acid oxidation and autophagy promote endoxifen resistance and counter the effect of AKT inhibition in ER-positive breast cancer cells.
    Lei Duan, Sarah Calhoun, Daeun Shim, Ricardo E Perez, Lothar A Blatter, Carl G Maki.
      Tamoxifen (TAM) is the first-line endocrine therapy for estrogen receptor-positive (ER+) breast cancer (BC). However, acquired resistance occurs in ∼50% cases. Meanwhile, although the PI3K/AKT/mTOR pathway is a viable target for treatment of endocrine therapy-refractory patients, complex signaling feedback loops exist, which can counter the effectiveness of inhibitors of this pathway. Here, we analyzed signaling pathways and metabolism in ER+ MCF7 BC cell line and their TAM-resistant derivatives that are co-resistant to endoxifen using immunoblotting, quantitative polymerase chain reaction, and the Agilent Seahorse XF Analyzer. We found that activation of AKT and the energy-sensing kinase AMPK was increased in TAM and endoxifen-resistant cells. Furthermore, ERRα/PGC-1β and their target genes MCAD and CPT-1 were increased and regulated by AMPK, which coincided with increased fatty acid oxidation (FAO) and autophagy in TAM-resistant cells. Inhibition of AKT feedback activates AMPK and ERRα/PGC-1β-MCAD/CPT-1 with a consequent increase in FAO and autophagy that counters the therapeutic effect of endoxifen and AKT inhibitors. Therefore, our results indicate increased activation of AKT and AMPK with metabolic reprogramming and increased autophagy in TAM-resistant cells. Simultaneous inhibition of AKT and FAO/autophagy is necessary to fully sensitize resistant cells to endoxifen.
    Keywords:  AKT; AMPK; autophagy; endoxifen; fatty acid oxidation
    DOI:  https://doi.org/10.1093/jmcb/mjab018
  25. Front Cell Dev Biol. 2021 ;9 644851
    Autophagy Mediates Leptin-Induced Migration and ERK Activation in Breast Cancer Cells.
    Alin García-Miranda, Karen Aylín Solano-Alcalá, José Benito Montes-Alvarado, Arely Rosas-Cruz, Julio Reyes-Leyva, Napoleón Navarro-Tito, Paola Maycotte, Eduardo Castañeda-Saucedo.
      Autophagy is an intracellular recycling process active in eukaryotic cells that involves the formation of an autophagosome which delivers cytoplasmic components to the lysosome for degradation. This process occurs at low rates under basal conditions, but it can be induced by diverse types of stress such as starvation, hypoxia, metabolic disorders or in response to hormones, including leptin. Leptin is considered a pro-tumorigenic protein whose circulating levels have been related to bad prognosis in obese breast cancer patients. It has been recently demonstrated that leptin can induce autophagy in cancer cell lines from different tissues, suggesting that autophagy could modulate the pro-tumorigenic effects associated with leptin. In this study, the role of autophagy in leptin-induced proliferation, migration, apoptosis and ERK phosphorylation in breast cancer cell lines was evaluated. Although leptin differentially induced autophagy in the breast cancer cell lines tested, autophagy inhibition reduced leptin-induced cell proliferation in MCF7 cells and decreased cell migration, ERK activation, and impaired morphological changes in both cell lines. Our data demonstrates an important role for basal autophagy or leptin-induced autophagy in leptin-induced migration and ERK phosphorylation in breast cancer cell lines, suggesting a potential use for the inhibition of autophagy in breast cancer associated with obesity.
    Keywords:  ERK; autophagy; breast cancer; leptin; migration; proliferation
    DOI:  https://doi.org/10.3389/fcell.2021.644851
  26. Aging (Albany NY). 2021 Mar 26. 13
    Host-commensal interaction promotes health and lifespan in Caenorhabditis elegans through the activation of HLH-30/TFEB-mediated autophagy.
    Miroslav Dinić, Marija Herholz, Uroš Kačarević, Dušan Radojević, Katarina Novović, Jelena Đokić, Aleksandra Trifunović, Nataša Golić.
      Gut homeostasis is maintained by the close interaction between commensal intestinal microbiota and the host, affecting the most complex physiological processes, such as aging. Some commensal bacteria with the potential to promote healthy aging arise as attractive candidates for the development of pro-longevity probiotics. Here, we showed that heat-inactivated human commensal Lactobacillus fermentum BGHV110 (BGHV110) extends the lifespan of Caenorhabditis elegans and improves age-related physiological features, including locomotor function and lipid metabolism. Mechanistically, we found that BGHV110 promotes HLH-30/TFEB-dependent autophagy to delay aging, as longevity assurance was completely abolished in the mutant lacking HLH-30, a major autophagy regulator in C. elegans. Moreover, we observed that BGHV110 partially decreased the content of lipid droplets in an HLH-30-dependent manner and, at the same time, slightly increased mitochondrial activity. In summary, this study demonstrates that specific factors from commensal bacteria can be used to exploit HLH-30/TFEB-mediated autophagy in order to promote longevity and fitness of the host.
    Keywords:  Caenorhabditis elegans; HLH-30; Lactobacillus fermentum; aging; autophagy
    DOI:  https://doi.org/10.18632/aging.202885
  27. Front Cell Dev Biol. 2021 ;9 637084
    Regulation of Age-Related Protein Toxicity.
    Anita Pras, Ellen A A Nollen.
      Proteome damage plays a major role in aging and age-related neurodegenerative diseases. Under healthy conditions, molecular quality control mechanisms prevent toxic protein misfolding and aggregation. These mechanisms include molecular chaperones for protein folding, spatial compartmentalization for sequestration, and degradation pathways for the removal of harmful proteins. These mechanisms decline with age, resulting in the accumulation of aggregation-prone proteins that are harmful to cells. In the past decades, a variety of fast- and slow-aging model organisms have been used to investigate the biological mechanisms that accelerate or prevent such protein toxicity. In this review, we describe the most important mechanisms that are required for maintaining a healthy proteome. We describe how these mechanisms decline during aging and lead to toxic protein misassembly, aggregation, and amyloid formation. In addition, we discuss how optimized protein homeostasis mechanisms in long-living animals contribute to prolonging their lifespan. This knowledge might help us to develop interventions in the protein homeostasis network that delay aging and age-related pathologies.
    Keywords:  aggregation; aging; amyloid; phase separation; protein homeostasis; protein quality control
    DOI:  https://doi.org/10.3389/fcell.2021.637084
  28. Cell Biosci. 2021 Mar 20. 11(1): 56
    Is targeting autophagy mechanism in cancer a good approach? The possible double-edge sword effect.
    Su Min Lim, Ezanee Azlina Mohamad Hanif, Siok-Fong Chin.
      Autophagy is a conserved cellular process required to maintain homeostasis. The hallmark of autophagy is the formation of a phagophore that engulfs cytosolic materials for degradation and recycling to synthesize essential components. Basal autophagy is constitutively active under normal conditions and it could be further induced by physiological stimuli such as hypoxia, nutrient starvation, endoplasmic reticulum stress,energy depletion, hormonal stimulation and pharmacological treatment. In cancer, autophagy is highly context-specific depending on the cell type, tumour microenvironment, disease stage and external stimuli. Recently, the emerging role of autophagy as a double-edged sword in cancer has gained much attention. On one hand, autophagy suppresses malignant transformation by limiting the production of reactive oxygen species and DNA damage during tumour development. Subsequently, autophagy evolved to support the survival of cancer cells and promotes the tumourigenicity of cancer stem cells at established sites. Hence, autophagy is an attractive target for cancer therapeutics and researchers have been exploiting the use of autophagy modulators as adjuvant therapy. In this review, we present a summary of autophagy mechanism and controlling pathways, with emphasis on the dual-role of autophagy (double-edged sword) in cancer. This is followed by an overview of the autophagy modulation for cancer treatment and is concluded by a discussion on the current perspectives and future outlook of autophagy exploitation for precision medicine.
    Keywords:  Autophagy; Cancer; LRG1; Precision Medicine; Treatment
    DOI:  https://doi.org/10.1186/s13578-021-00570-z
  29. Neuron. 2021 Mar 18. pii: S0896-6273(21)00156-2. [Epub ahead of print]
    Akt-mTOR hypoactivity in bipolar disorder gives rise to cognitive impairments associated with altered neuronal structure and function.
    Amanda M Vanderplow, Andrew L Eagle, Bailey A Kermath, Kathryn J Bjornson, Alfred J Robison, Michael E Cahill.
      The Akt family of kinases exerts many of its cellular effects via the activation of the mammalian target of rapamycin (mTOR) kinase through a series of intermediary proteins. Multiple lines of evidence have identified Akt-family kinases as candidate schizophrenia and bipolar disorder genes. Although dysfunction of the prefrontal cortex (PFC) is a key feature of both schizophrenia and bipolar disorder, no studies have comprehensively assessed potential alterations in Akt-mTOR pathway activity in the PFC of either disorder. Here, we examined the activity and expression profile of key proteins in the Akt-mTOR pathway in bipolar disorder and schizophrenia homogenates from two different PFC subregions. Our findings identify reduced Akt-mTOR PFC signaling in a subset of bipolar disorder subjects. Using a reverse-translational approach, we demonstrated that Akt hypofunction in the PFC is sufficient to give rise to key cognitive phenotypes that are paralleled by alterations in synaptic connectivity and function.
    Keywords:  akt; autophagy; bipolar disorder; cognition; dendritic spine; mTOR; memory; prefrontal cortex; synapse; ulk1
    DOI:  https://doi.org/10.1016/j.neuron.2021.03.008
  30. Clin Exp Pharmacol Physiol. 2021 Mar 22.
    Targeting Autophagy in Neurodegenerative Diseases: from Molecular Mechanisms to Clinical Therapeutics.
    Amir Ajoolabady, Hamid Aslkhodapasandhokmabad, Nils Henninger, Laurie J Demillard, Masoud Nikanfar, Alireza Nourazarian, Jun Ren.
      Many neurodegenerative diseases are associated with pathological aggregation of proteins in neurons. Autophagy is a natural self-cannibalization process that can act as a powerful mechanism to remove aged and damaged organelles as well as protein aggregates. It has been shown that promoting autophagy can attenuate or delay neurodegeneration by removing protein aggregates. In this paper, we will review the role of autophagy in Alzheimer's disease (AD), Parkinson's Disease (PD), and Huntington's Disease (HD) and discuss opportunities and challenges of targeting autophagy as a potential therapeutic avenue for treatment of these common neurodegenerative diseases.
    Keywords:  Alzheimer’s disease (AD); Parkinson’s Disease (PD); and Huntington’s Disease (HD); autophagy; neurodegenerative disease
    DOI:  https://doi.org/10.1111/1440-1681.13500
  31. Mol Cell. 2021 Mar 11. pii: S1097-2765(21)00167-2. [Epub ahead of print]
    Global phosphoproteomics pinpoints uncharted Gcn2-mediated mechanisms of translational control.
    Ladislav Dokládal, Michael Stumpe, Benjamin Pillet, Zehan Hu, Guillermo Miguel Garcia Osuna, Dieter Kressler, Jörn Dengjel, Claudio De Virgilio.
      The conserved Gcn2 protein kinase mediates cellular adaptations to amino acid limitation through translational control of gene expression that is exclusively executed by phosphorylation of the α-subunit of the eukaryotic translation initiation factor 2 (eIF2α). Using quantitative phosphoproteomics, however, we discovered that Gcn2 targets auxiliary effectors to modulate translation. Accordingly, Gcn2 also phosphorylates the β-subunit of the trimeric eIF2 G protein complex to promote its association with eIF5, which prevents spontaneous nucleotide exchange on eIF2 and thereby restricts the recycling of the initiator methionyl-tRNA-bound eIF2-GDP ternary complex in amino-acid-starved cells. This mechanism contributes to the inhibition of translation initiation in parallel to the sequestration of the nucleotide exchange factor eIF2B by phosphorylated eIF2α. Gcn2 further phosphorylates Gcn20 to antagonize, in an inhibitory feedback loop, the formation of the Gcn2-stimulatory Gcn1-Gcn20 complex. Thus, Gcn2 plays a substantially more intricate role in controlling translation initiation than hitherto appreciated.
    Keywords:  Gcn2; Gcn20; TORC1; amino acid starvation; eIF2; eIF5; eukaryotic initiation factor 2; eukaryotic initiation factor 5; general control nonderepressible 2; target of rapamycin complex 1; translation initiation
    DOI:  https://doi.org/10.1016/j.molcel.2021.02.037
  32. Front Oncol. 2021 ;11 599124
    Inhibition of HMGB1 Suppresses Hepatocellular Carcinoma Progression via HIPK2-Mediated Autophagic Degradation of ZEB1.
    Wei Zhu, Jun Li, Yuheng Zhang, Zhengyi Zhu, Hanyi Liu, Yunzhen Lin, Anyin Hu, Jingchao Zhou, Haozhen Ren, Xiaolei Shi.
      Autophagy is a conserved catabolic process maintaining cellular homeostasis and reportedly plays a critical role in tumor progression. Accumulating data show that autophagic activity is inhibited in hepatocellular carcinoma. However, the underlying molecular basis of impaired autophagy in HCC remains unclear. In this study, we revealed that autophagic activity was suppressed by HMGB1 in a HIPK2-dependent way. Targeting HMGB1 could inhibit the degradation of HIPK2, as a result of which, autophagic degradation of ZEB1 was enhanced by reprogramming glucose metabolism/AMPK/mTOR axis. Moreover, we demonstrated that selectively degradation of ZEB1 was responsible for HCC growth inhibition in HMGB1 deficient cells. Lastly, we found the combination therapy of HMGB1 inhibitor and rapamycin achieved a better anti-HCC effect. These results demonstrate that impaired autophagy is controlled by HMGB1 and targeting HMGB1 could suppress HCC progression via HIPK2-mediated autophagic degradation of ZEB1.
    Keywords:  autophagy; epithelial-mesenchymal transition (EMT); glucose metabolism; hepatocellular carcinoma (HCC); high-mobility group box 1 (HMGB1)
    DOI:  https://doi.org/10.3389/fonc.2021.599124
  33. Nutr Cancer. 2021 Mar 24. 1-10
    Luteolin Induces Apoptosis and Autophagy in HCT116 Colon Cancer Cells via p53-Dependent Pathway.
    Ho Soo Yoo, Sae Bom Won, Young Hye Kwon.
      Although a dietary phytochemical luteolin has been shown to regulate various anticancer mechanisms, a role of luteolin in autophagy regulation is mostly unidentified. Here, we investigated whether luteolin exhibits its anticancer effects by induction of apoptosis and autophagy in a p53-dependent manner in colon cancer cells. Cell viability was determined using trypan blue exclusion test. The expressions of proteins and mRNAs were measured by immunoblotting and reverse transcription polymerase chain reaction, respectively. Luteolin at 10 - 20 μM induced cytotoxicity in p53 wild-type HCT116 colon cancer cells but not in p53 mutant HT-29 cells and normal colon cells. Luteolin exhibited its anticancer effect by increasing p53 phosphorylation and p53 target gene expression, leading to apoptosis and cell cycle arrest in HCT116 cells. We identified that luteolin can induce autophagy in p53 wild-type cells but not in p53 mutant cells, suggesting that luteolin-induced autophagy is p53-dependent; however, chloroquine-mediated inhibition of autophagy did not alter cytotoxicity and apoptosis of cells treated with luteolin. In conclusion, the present data showed that luteolin inhibits the growth of HCT116 colon cancer cells through p53-dependent regulation of apoptosis and cell cycle arrest regardless of the induction of autophagy.
    DOI:  https://doi.org/10.1080/01635581.2021.1903947
  34. Hum Mol Genet. 2021 Mar 22. pii: ddab080. [Epub ahead of print]
    Salvage NAD+ Biosynthetic Pathway Enzymes Moonlight as Molecular Chaperones to Protect Against Proteotoxicity.
    Meredith Pinkerton, Andrea Ruetenik, Viktoriia Bazylianska, Eva Nyvltova, Antoni Barrientos.
      Human neurodegenerative proteinopathies are disorders associated with abnormal protein depositions in brain neurons. They include polyglutamine (polyQ) conditions such as Huntington's disease (HD) and α-synucleinopathies such as Parkinson's disease (PD). Overexpression of NMNAT/Nma1, an enzyme in the NAD+ biosynthetic salvage pathway, acts as an efficient suppressor of proteotoxicities in yeast, fly, and mouse models. Screens in yeast models of HD and PD allowed us to identify three additional enzymes of the same pathway that achieve similar protection against proteotoxic stress: Npt1, Pnc1, and Qns1. The mechanism by which these proteins maintain proteostasis has not been identified. Here, we report that their ability to maintain proteostasis in yeast models of HD and PD is independent of their catalytic activity and does not require cellular protein quality control systems such as the proteasome or autophagy. Furthermore, we show that, under proteotoxic stress, the four proteins are recruited as molecular chaperones with holdase and foldase activities. The NAD+ salvage proteins act by preventing misfolding and, together with the Hsp90 chaperone, promoting the refolding of extended polyQ domains and α-synuclein (α-Syn). Our results illustrate the existence of an evolutionarily conserved strategy of repurposing or moonlighting housekeeping enzymes under stress conditions to maintain proteostasis. We conclude that the entire salvage NAD+ biosynthetic pathway links NAD+ metabolism and proteostasis and emerges as a target for therapeutics to combat age-associated neurodegenerative proteotoxicities.
    DOI:  https://doi.org/10.1093/hmg/ddab080
  35. Biochem Soc Trans. 2021 Mar 26. pii: BST20190236. [Epub ahead of print]
    Parkinson's disease and mitophagy: an emerging role for LRRK2.
    Francois Singh, Ian G Ganley.
      Parkinson's disease (PD) is a progressive neurodegenerative disorder that affects around 2% of individuals over 60 years old. It is characterised by the loss of dopaminergic neurons in the substantia nigra pars compacta of the midbrain, which is thought to account for the major clinical symptoms such as tremor, slowness of movement and muscle stiffness. Its aetiology is poorly understood as the physiological and molecular mechanisms leading to this neuronal loss are currently unclear. However, mitochondrial and lysosomal dysfunction seem to play a central role in this disease. In recent years, defective mitochondrial elimination through autophagy, termed mitophagy, has emerged as a potential contributing factor to disease pathology. PINK1 and Parkin, two proteins mutated in familial PD, were found to eliminate mitochondria under distinct mitochondrial depolarisation-induced stress. However, PINK1 and Parkin are not essential for all types of mitophagy and such pathways occur in most cell types and tissues in vivo, even in the absence of overt mitochondrial stress - so-called basal mitophagy. The most common mutation in PD, that of glycine at position 2019 to serine in the protein kinase LRRK2, results in increased activity and this was recently shown to disrupt basal mitophagy in vivo. Thus, different modalities of mitophagy are affected by distinct proteins implicated in PD, suggesting impaired mitophagy may be a common denominator for the disease. In this short review, we discuss the current knowledge about the link between PD pathogenic mutations and mitophagy, with a particular focus on LRRK2.
    Keywords:  Parkinsons disease; autophagy; leucine-rich repeat kinase; mitophagy
    DOI:  https://doi.org/10.1042/BST20190236
  36. Acta Trop. 2021 Mar 17. pii: S0001-706X(21)00069-3. [Epub ahead of print] 105890
    Leishmania (V.) braziliensis infection promotes macrophage autophagy by a LC3B-dependent and BECLIN1-independent mechanism.
    Thabata Lopes Alberto Duque, Thamires Christinne de Souza Lopes Cruz Serrão, Antônio José da Silva Gonçalves, Eduardo Fonseca Pinto, Manoel Paes de Oliveira Neto, Claude Pirmez, Luiza de Oliveira Ramos Pereira, Rubem Figueiredo Sadok Menna-Barreto.
      Leishmania (Viannia) braziliensis is one of the main etiological agents of tegumentary leishmaniasis in Latin America. The establishment of a successful infection in host cells requires several key events including phagocytosis, phagolysosomal maturation impairment, and parasite replication. Autophagy is accountable for the physiological turnover of cellular organelles, degradation of macromolecular structures, and pathogen elimination. In many cases, autophagy control leads to a successful infection, both impairing pathogen elimination or providing nutrients. Here, we have investigated the relationship between autophagy and L. braziliensis infection. We observed that BECLIN1 expression was upregulated early on infection in both in vitro macrophage cultures and biopsies of cutaneous lesions from L. braziliensis infected patients. On the other hand, LC3B expression was downregulated in cutaneous lesions biopsies. A transient pattern of LC3+ cells was observed along L. braziliensis infection, but the number of LC3 puncta did not vary. Additionally, autophagy induction, with rapamycin treatment or through starvation, reduced infection. As expected, rapamycin increased the percentage of LC3+ cells and the number of puncta, but the presence of parasite restricted that effect, indicating LC3-associated autophagy impairment by L. braziliensis. Finally, silencing LC3B but not BECLIN1 promoted infection, confirming BECLIN1 independent and LC3B-related control by the parasite. Taken together, these data indicate macrophage autophagic machinery manipulation by L. braziliensis, resulting in successful establishment and survival into the host cell.
    Keywords:  Autophagy; BECLIN1; LC3B; Leishmania spp.; Leishmaniasis; Macrophages
    DOI:  https://doi.org/10.1016/j.actatropica.2021.105890
  37. Aging Cell. 2021 Mar 21. e13347
    VPS35 D620N knockin mice recapitulate cardinal features of Parkinson's disease.
    Mengyue Niu, Fanpeng Zhao, Karina Bondelid, Sandra L Siedlak, Sandy Torres, Hisashi Fujioka, Wenzhang Wang, Jun Liu, Xiongwei Zhu.
      D620N mutation in the vacuolar protein sorting 35 ortholog (VPS35) gene causes late-onset, autosomal dominant familial Parkinson's disease (PD) and contributes to idiopathic PD. However, how D620N mutation leads to PD-related deficits in vivo remains unclear. In the present study, we thoroughly characterized the biochemical, pathological, and behavioral changes of a VPS35 D620N knockin (KI) mouse model with chronic aging. We reported that this VPS35 D620N KI model recapitulated a spectrum of cardinal features of PD at 14 months of age which included age-dependent progressive motor deficits, significant changes in the levels of dopamine (DA) and DA metabolites in the striatum, and robust neurodegeneration of the DA neurons in the SNpc and DA terminals in the striatum, accompanied by increased neuroinflammation, and accumulation and aggregation of α-synuclein in DA neurons. Mechanistically, D620N mutation induced mitochondrial fragmentation and dysfunction in aged mice likely through enhanced VPS35-DLP1 interaction and increased turnover of mitochondrial DLP1 complexes in vivo. Finally, the VPS35 D620N KI mice displayed greater susceptibility to MPTP-mediated degeneration of nigrostriatal pathway, indicating that VPS35 D620N mutation increased vulnerability of DA neurons to environmental toxins. Overall, this VPS35 D620N KI mouse model provides a powerful tool for future disease modeling and pharmacological studies of PD. Our data support the involvement of VPS35 in the development of α-synuclein pathology in vivo and revealed the important role of mitochondrial fragmentation/dysfunction in the pathogenesis of VPS35 D620N mutation-associated PD in vivo.
    Keywords:  Parkinson's disease; VPS35; mitochondria dynamics; neurodegeneration; α-synuclein
    DOI:  https://doi.org/10.1111/acel.13347
  38. Curr Biol. 2021 Mar 18. pii: S0960-9822(21)00305-5. [Epub ahead of print]
    Increased LRRK2 kinase activity alters neuronal autophagy by disrupting the axonal transport of autophagosomes.
    C Alexander Boecker, Juliet Goldsmith, Dan Dou, Gregory G Cajka, Erika L F Holzbaur.
      Parkinson's disease-causing mutations in the leucine-rich repeat kinase 2 (LRRK2) gene hyperactivate LRRK2 kinase activity and cause increased phosphorylation of Rab GTPases, important regulators of intracellular trafficking. We found that the most common LRRK2 mutation, LRRK2-G2019S, dramatically reduces the processivity of autophagosome transport in neurons in a kinase-dependent manner. This effect was consistent across an overexpression model, neurons from a G2019S knockin mouse, and human induced pluripotent stem cell (iPSC)-derived neurons gene edited to express the G2019S mutation, and the effect was reversed by genetic or pharmacological inhibition of LRRK2. Furthermore, LRRK2 hyperactivation induced by overexpression of Rab29, a known activator of LRRK2 kinase, disrupted autophagosome transport to a similar extent. Mechanistically, we found that hyperactive LRRK2 recruits the motor adaptor JNK-interacting protein 4 (JIP4) to the autophagosomal membrane, inducing abnormal activation of kinesin that we propose leads to an unproductive tug of war between anterograde and retrograde motors. Disruption of autophagosome transport correlated with a significant defect in autophagosome acidification, suggesting that the observed transport deficit impairs effective degradation of autophagosomal cargo in neurons. Our results robustly link increased LRRK2 kinase activity to defects in autophagosome transport and maturation, further implicating defective autophagy in the pathogenesis of Parkinson's disease.
    Keywords:  JIP4; LRRK2; Parkinson's disease; Rab29; autophagy; axonal transport
    DOI:  https://doi.org/10.1016/j.cub.2021.02.061
  39. Cell Mol Gastroenterol Hepatol. 2021 Mar 22. pii: S2352-345X(21)00059-X. [Epub ahead of print]
    DJ-1 deficiency in hepatocytes improves liver ischemia-reperfusion injury by enhancing mitophagy.
    Min Xu, Hualian Hang, Miao Huang, Jichang Li, Dongwei Xu, Junzhe Jiao, Fang Wang, Hailong Wu, Xuehua Sun, Jinyang Gu, Xiaoni Kong, Yueqiu Gao.
      BACKGROUND & AIMS: DJ-1 is universally expressed in various tissues and organs, and involved in the physiological processes in various liver diseases. However, the role of DJ-1 in liver ischemia-reperfusion (I/R) injury is largely unknown.METHODS: In this study, we first examined the DJ-1 expression changes in the liver tissues of mice and clinical donor post hepatic I/R by both qPCR and western blotting assays. And then we investigate the role of DJ-1 in I/R injury by employing a murine liver I/R model.
    RESULTS: We demonstrate that DJ-1 down-regulation in both human and mouse liver tissues in response to I/R injury, and Dj-1 deficiency in hepatocytes but not in myeloid cells could significantly ameliorate I/R induced liver injury and inflammatory responses. This hepatoprotective effects was dependent on enhanced autophagy in Dj-1 knockout mice, because inhibition of autophagy by 3-Methyladenine (3-MA) and chloroquine (CQ) could reverse the protective effect on hepatic I/R injury in Dj-1 knockout mice CONCLUSION: Dj-1 deficiency in hepatocytes significantly enhanced mitochondrial accumulation and protein stability of PARKIN, which in turn promotes the onset of mitophagy resulting in elevated clearance of damaged mitochondria during I/R injury.
    Keywords:  DJ-1; PARKIN; liver I/R injury; mitophagy
    DOI:  https://doi.org/10.1016/j.jcmgh.2021.03.007
  40. Aging (Albany NY). 2021 Mar 21. 13
    Evolution of mammalian longevity: age-related increase in autophagy in bats compared to other mammals.
    Joanna Kacprzyk, Andrea G Locatelli, Graham M Hughes, Zixia Huang, Michael Clarke, Vera Gorbunova, Carlotta Sacchi, Gavin S Stewart, Emma C Teeling.
      Autophagy maintains cellular homeostasis and its dysfunction has been implicated in aging. Bats are the longest-lived mammals for their size, but the molecular mechanisms underlying their extended healthspan are not well understood. Here, drawing on >8 years of mark-recapture field studies, we report the first longitudinal analysis of autophagy regulation in bats. Mining of published population level aging blood transcriptomes (M. myotis, mouse and human) highlighted a unique increase of autophagy related transcripts with age in bats, but not in other mammals. This bat-specific increase in autophagy transcripts was recapitulated by the western blot determination of the autophagy marker, LC3II/I ratio, in skin primary fibroblasts (Myotis myotis, Pipistrellus kuhlii, mouse), that also showed an increase with age in both bat species. Further phylogenomic selection pressure analyses across eutherian mammals (n=70 taxa; 274 genes) uncovered 10 autophagy-associated genes under selective pressure in bat lineages. These molecular adaptations potentially mediate the exceptional age-related increase of autophagy signalling in bats, which may contribute to their longer healthspans.
    Keywords:  aging; autophagy; bats; blood mRNA; phylogenomics
    DOI:  https://doi.org/10.18632/aging.202852
  41. Nat Metab. 2021 Mar;3(3): 428-441
    STK3/STK4 signalling in adipocytes regulates mitophagy and energy expenditure.
    Yoon Keun Cho, Yeonho Son, Abhirup Saha, Doeun Kim, Cheoljun Choi, Minsu Kim, Ji-Hyun Park, Hyeonyeong Im, Juhyeong Han, Kyungmin Kim, Young-Suk Jung, Jeanho Yun, Eun Ju Bae, Je Kyung Seong, Mi-Ock Lee, Sangkyu Lee, James G Granneman, Yun-Hee Lee.
      Obesity reduces adipocyte mitochondrial function, and expanding adipocyte oxidative capacity is an emerging strategy to improve systemic metabolism. Here, we report that serine/threonine-protein kinase 3 (STK3) and STK4 are key physiological suppressors of mitochondrial capacity in brown, beige and white adipose tissues. Levels of STK3 and STK4, kinases in the Hippo signalling pathway, are greater in white than brown adipose tissues, and levels in brown adipose tissue are suppressed by cold exposure and greatly elevated by surgical denervation. Genetic inactivation of Stk3 and Stk4 increases mitochondrial mass and function, stabilizes uncoupling protein 1 in beige adipose tissue and confers resistance to metabolic dysfunction induced by high-fat diet feeding. Mechanistically, STK3 and STK4 increase adipocyte mitophagy in part by regulating the phosphorylation and dimerization status of the mitophagy receptor BNIP3. STK3 and STK4 expression levels are elevated in human obesity, and pharmacological inhibition improves metabolic profiles in a mouse model of obesity, suggesting STK3 and STK4 as potential targets for treating obesity-related diseases.
    DOI:  https://doi.org/10.1038/s42255-021-00362-2
  42. Autophagy. 2021 Mar 22. 1-14
    Modulation of the IGF1R-MTOR pathway attenuates motor neuron toxicity of human ALS SOD1G93A astrocytes.
    Veronica Granatiero, Nicole M Sayles, Angela M Savino, Csaba Konrad, Michael G Kharas, Hibiki Kawamata, Giovanni Manfredi.
      ALS (amyotrophic lateral sclerosis), the most common motor neuron disease, causes muscle denervation and rapidly fatal paralysis. While motor neurons are the most affected cells in ALS, studies on the pathophysiology of the disease have highlighted the importance of non-cell autonomous mechanisms, which implicate astrocytes and other glial cells. In ALS, subsets of reactive astrocytes lose their physiological functions and become toxic for motor neurons, thereby contributing to disease pathogenesis. Evidence of astrocyte contribution to disease pathogenesis are well established in cellular and animal models of familial ALS linked to mutant SOD1, where astrocytes promote motor neuron cell death. The mechanism underlying astrocytes reactivity in conditions of CNS injury have been shown to involve the MTOR pathway. However, the role of this conserved metabolic signaling pathway, and the potential therapeutic effects of its modulation, have not been investigated in ALS astrocytes. Here, we show elevated activation of the MTOR pathway in human-derived astrocytes harboring mutant SOD1, which results in inhibition of macroautophagy/autophagy, increased cell proliferation, and enhanced astrocyte reactivity. We demonstrate that MTOR pathway activation in mutant SOD1 astrocytes is due to post-transcriptional upregulation of the IGF1R (insulin like growth factor 1 receptor), an upstream positive modulator of the MTOR pathway. Importantly, inhibition of the IGF1R-MTOR pathway decreases cell proliferation and reactivity of mutant SOD1 astrocytes, and attenuates their toxicity to motor neurons. These results suggest that modulation of astrocytic IGF1R-MTOR pathway could be a viable therapeutic strategy in SOD1 ALS and potentially other neurological diseases.
    Keywords:  Astrocytes; EIF4EBP1; IGF1R; PPP; SOD1G93A; Torin1; ULK1; autophagy; motor neurons; mtor
    DOI:  https://doi.org/10.1080/15548627.2021.1899682
  43. Nat Commun. 2021 Mar 26. 12(1): 1920
    The oncogene AAMDC links PI3K-AKT-mTOR signaling with metabolic reprograming in estrogen receptor-positive breast cancer.
    Emily Golden, Rabab Rashwan, Eleanor A Woodward, Agustin Sgro, Edina Wang, Anabel Sorolla, Charlene Waryah, Wan Jun Tie, Elisabet Cuyàs, Magdalena Ratajska, Iwona Kardaś, Piotr Kozlowski, Elizabeth K M Johnstone, Heng B See, Ciara Duffy, Jeremy Parry, Kim A Lagerborg, Piotr Czapiewski, Javier A Menendez, Adam Gorczyński, Bartosz Wasag, Kevin D G Pfleger, Christina Curtis, Bum-Kyu Lee, Jonghwan Kim, Joseph Cursons, Nathan J Pavlos, Wojciech Biernat, Mohit Jain, Andrew J Woo, Andrew Redfern, Pilar Blancafort.
      Adipogenesis associated Mth938 domain containing (AAMDC) represents an uncharacterized oncogene amplified in aggressive estrogen receptor-positive breast cancers. We uncover that AAMDC regulates the expression of several metabolic enzymes involved in the one-carbon folate and methionine cycles, and lipid metabolism. We show that AAMDC controls PI3K-AKT-mTOR signaling, regulating the translation of ATF4 and MYC and modulating the transcriptional activity of AAMDC-dependent promoters. High AAMDC expression is associated with sensitization to dactolisib and everolimus, and these PI3K-mTOR inhibitors exhibit synergistic interactions with anti-estrogens in IntClust2 models. Ectopic AAMDC expression is sufficient to activate AKT signaling, resulting in estrogen-independent tumor growth. Thus, AAMDC-overexpressing tumors may be sensitive to PI3K-mTORC1 blockers in combination with anti-estrogens. Lastly, we provide evidence that AAMDC can interact with the RabGTPase-activating protein RabGAP1L, and that AAMDC, RabGAP1L, and Rab7a colocalize in endolysosomes. The discovery of the RabGAP1L-AAMDC assembly platform provides insights for the design of selective blockers to target malignancies having the AAMDC amplification.
    DOI:  https://doi.org/10.1038/s41467-021-22101-7
  44. Life Sci. 2021 Mar 23. pii: S0024-3205(21)00337-4. [Epub ahead of print] 119352
    Autophagy-lysosomal signaling responses to heat stress in tenotomy-induced rat skeletal muscle atrophy.
    Muthita Hirunsai, Ratchakrit Srikuea.
      AIMS: The autophagy-lysosomal system plays a crucial role in maintaining muscle proteostasis. Excessive stimulation of the autophagic machinery is a major contributor to muscle atrophy induced by tendon transection. Hyperthermia is known to attenuate muscle protein loss during disuse conditions; however, little is known regarding the response of the autophagy pathway to heat stress following tenotomy-induced muscle atrophy. The purpose of this study was to evaluate whether heat stress would have a beneficial impact on the activation of autophagy in tenotomized soleus and plantaris muscles.MAIN METHODS: Male Wistar rats were divided into control, control plus heat stress, tenotomy, and tenotomy plus heat stress groups. The effects of tenotomy were evaluated at 8 and 14 days with heat treatment applied using thermal blankets (30 min. day-1, at 40.5-41.5 °C, for 7 days).
    KEY FINDINGS: Heat stress could normalize tenotomy-induced muscle loss and over-activation of autophagy-lysosomal signaling; this effect was evidently observed in soleus muscle tenotomized for 14 days. The autophagy-related proteins LC3B-II and LC3B-II/I tended to decrease, and lysosomal cathepsin L protein expression was significantly suppressed. While p62/SQSTM1 was not altered in response to intermittent heat exposure in tenotomized soleus muscle at day 14. Phosphorylation of the 4E-BP1 protein was significantly increased in tenotomized plantaris muscle; whereas heat stress had no impact on phosphorylation of Akt and FoxO3a proteins in both tenotomized muscles examined.
    SIGNIFICANCE: Our results provide evidence that heat stress associated attenuation of tenotomy-induced muscle atrophy is mediated through limiting over-activation of the autophagy-lysosomal pathway in oxidative and glycolytic muscles.
    Keywords:  4E-BP1; Cathepsin; FoxO3a; LC3B; Lysosome; Muscle wasting; Sequestosome
    DOI:  https://doi.org/10.1016/j.lfs.2021.119352
  45. Aging (Albany NY). 2021 Mar 21. 13
    Enhancing lifespan of budding yeast by pharmacological lowering of amino acid pools.
    Nathaniel L Hepowit, Jessica K A Macedo, Lyndsay E A Young, Ke Liu, Ramon C Sun, Jason A MacGurn, Robert C Dickson.
      The increasing prevalence of age-related diseases and resulting healthcare insecurity and emotional burden require novel treatment approaches. Several promising strategies seek to limit nutrients and promote healthy aging. Unfortunately, the human desire to consume food means this strategy is not practical for most people but pharmacological approaches might be a viable alternative. We previously showed that myriocin, which impairs sphingolipid synthesis, increases lifespan in Saccharomyces cerevisiae by modulating signaling pathways including the target of rapamycin complex 1 (TORC1). Since TORC1 senses cellular amino acids, we analyzed amino acid pools and identified 17 that are lowered by myriocin treatment. Studying the methionine transporter, Mup1, we found that newly synthesized Mup1 traffics to the plasma membrane and is stable for several hours but is inactive in drug-treated cells. Activity can be restored by adding phytosphingosine to culture medium thereby bypassing drug inhibition, thus confirming a sphingolipid requirement for Mup1 activity. Importantly, genetic analysis of myriocin-induced longevity revealed a requirement for the Gtr1/2 (mammalian Rags) and Vps34-Pib2 amino acid sensing pathways upstream of TORC1, consistent with a mechanism of action involving decreased amino acid availability. These studies demonstrate the feasibility of pharmacologically inducing a state resembling amino acid restriction to promote healthy aging.
    Keywords:  amino acid transport; lifespan; longevity; nutrient restriction; sphingolipids
    DOI:  https://doi.org/10.18632/aging.202849
  46. Dev Cell. 2021 Mar 19. pii: S1534-5807(21)00202-1. [Epub ahead of print]
    ER-phagy responses in yeast, plants, and mammalian cells and their crosstalk with UPR and ERAD.
    Maurizio Molinari.
      ER-phagy, literally endoplasmic reticulum (ER)-eating, defines the constitutive or regulated clearance of ER portions within metazoan endolysosomes or yeast and plant vacuoles. The advent of electron microscopy led to the first observations of ER-phagy over 60 years ago, but only recently, with the discovery of a set of regulatory proteins named ER-phagy receptors, has it been dissected mechanistically. ER-phagy receptors are activated by a variety of pleiotropic and ER-centric stimuli. They promote ER fragmentation and engage luminal, membrane-bound, and cytosolic factors, eventually driving lysosomal clearance of select ER domains along with their content. After short historical notes, this review introduces the concept of ER-phagy responses (ERPRs). ERPRs ensure lysosomal clearance of ER portions expendable during nutrient shortage, nonfunctional, present in excess, or containing misfolded proteins. They cooperate with unfolded protein responses (UPRs) and with ER-associated degradation (ERAD) in determining ER size, function, and homeostasis.
    Keywords:  ER-associated degradation (ERAD); ER-phagy; ER-phagy response (ERPR); ER-to-lysosome-associated; autophagosome; autophagy; degradation (ERLAD); endolysosome; lysosome; recov-ER-phagy; unfolded proteins response (UPR); vacuole
    DOI:  https://doi.org/10.1016/j.devcel.2021.03.005
  47. Exp Biol Med (Maywood). 2021 Mar 23. 15353702211002136
    Arbutin ameliorates glucocorticoid-induced osteoporosis through activating autophagy in osteoblasts.
    Yiqi Zhang, Mingyang Li, Ziyun Liu, Qin Fu.
      Chronic long-term glucocorticoid use causes osteoporosis partly by interrupting osteoblast homeostasis and exacerbating bone loss. Arbutin, a natural hydroquinone glycoside, has been reported to have biological activities related to the differentiation of osteoblasts and osteoclasts. However, the role and underlying mechanism of arbutin in glucocorticoid-induced osteoporosis are elusive. In this study, we demonstrated that arbutin administration ameliorated osteoporotic disorders in glucocorticoid dexamethasone (Dex)-induced mouse model, including attenuating the loss of bone mass and trabecular microstructure, promoting bone formation, suppressing bone resorption, and activating autophagy in bone tissues. Furthermore, Dex-stimulated mouse osteoblastic MC3T3-E1 cells were treated with arbutin. Arbutin treatment rescued Dex-induced repression of osteoblast differentiation and mineralization, the downregulation of osteogenic gene expression, reduced autophagic marker expression, and decreased autophagic puncta formation. The application of autophagy inhibitor 3-MA decreased autophagy, differentiation, and mineralization of MC3T3-E1 cells triggered by arbutin. Taken together, our findings suggest that arbutin treatment fends off glucocorticoid-induced osteoporosis, partly through promoting differentiation and mineralization of osteoblasts by autophagy activation.
    Keywords:  Arbutin; autophagy; dexamethasone; osteoblasts; osteoporosis
    DOI:  https://doi.org/10.1177/15353702211002136
  48. Nat Rev Cancer. 2021 Mar 23.
    Autophagy in tumour immunity and therapy.
    Houjun Xia, Douglas R Green, Weiping Zou.
      Autophagy is a regulated mechanism that removes unnecessary or dysfunctional cellular components and recycles metabolic substrates. In response to stress signals in the tumour microenvironment, the autophagy pathway is altered in tumour cells and immune cells - thereby differentially affecting tumour progression, immunity and therapy. In this Review, we summarize our current understanding of the immunologically associated roles and modes of action of the autophagy pathway in cancer progression and therapy, and discuss potential approaches targeting autophagy to enhance antitumour immunity and improve the efficacy of current cancer therapy.
    DOI:  https://doi.org/10.1038/s41568-021-00344-2
  49. Cell Death Differ. 2021 Mar 22.
    PARsylated transcription factor EB (TFEB) regulates the expression of a subset of Wnt target genes by forming a complex with β-catenin-TCF/LEF1.
    Soyoung Kim, Gahyeon Song, Taebok Lee, Minseong Kim, Jeongrae Kim, Hyeryun Kwon, Jiyoung Kim, Wonyoung Jeong, Ukjin Lee, Chaebin Na, Sangwon Kang, Wantae Kim, Je Kyung Seong, Eek-Hoon Jho.
      Wnt signaling is mainly transduced by β-catenin via regulation of the β-catenin destruction complex containing Axin, APC, and GSK3β. Transcription factor EB (TFEB) is a well-known master regulator of autophagy and lysosomal biogenesis processes. TFEB's nuclear localization and transcriptional activity are also regulated by various upstream signals. In this study, we found that Wnt signaling induces the nuclear localization of TFEB and the expression of Wnt target genes is regulated by TFEB-β-catenin-TCF/LEF1 as well as β-catenin-TCF/LEF1 complexes. Our biochemical data revealed that TFEB is a part of the β-catenin destruction complex, and destabilization of the destruction complex by knockdown of either Axin or APC causes nuclear localization of TFEB. Interestingly, RNA-sequencing analysis revealed that about 27% of Wnt3a-induced genes were TFEB dependent. However, these "TFEB mediated Wnt target genes" were different from TFEB target genes involved in autophagy and lysosomal biogenesis processes. Mechanistically, we found that Tankyrase (TNKS) PARsylates TFEB with Wnt ON signaling, and the nuclear localized PARsylated TFEB forms a complex with β-catenin-TCF/LEF1 to induce the "TFEB mediated Wnt target genes". Finally, we found that in various types of cancer, the levels of TFEB mediated Wnt target genes exhibit strong correlations with the level of Axin2, which represents the activity of Wnt signaling. Overall, our data suggest that Wnt signaling induces the expression of a subset of genes that are distinct from previously known genes regulated by the β-catenin-TCF/LEF1 complex or TFEB, by forming a transcription factor complex consisting of PARsylated TFEB and β-catenin-TCF/LEF1.
    DOI:  https://doi.org/10.1038/s41418-021-00770-7
  50. Biochem Biophys Res Commun. 2021 Mar 17. pii: S0006-291X(21)00417-4. [Epub ahead of print]552 78-83
    The upregulation of Ulk1-dependent autophagy does not require the p53 activity in mouse embryonic stem cells.
    Mikhail L Vorobev, Bashar A Alhasan, Irina I Suvorova.
      Autophagy is known to play a critical role in the early stages of embryogenesis including the formation of blastocyst. The existence of p53 protein-deficient mice may identify that p53 is not indispensable for the activation of autophagy in pluripotent cells derived from the inner cell mass of the blastocyst. We utilized a p53-knockout (KO) mouse embryonic stem cell (mESC) line to investigate the contribution of p53 in autophagy. We showed that lack of p53 has no effect on cell pluripotency but significantly hinders the differentiation process induced by retinoic acid. Using MRT68921, we revealed that Ulk1-dependent autophagy is activated in response to serum deprivation despite the deletion of p53 in mESCs. However, under retinoic acid-induced differentiation, the accumulation of autophagosomes and lysosomes is impaired in p53 KO mESCs, indicating a critical role of p53 in the regulation of autophagy upon differentiation.
    Keywords:  Autophagy; Differentiation; Embryonic stem cells; Pluripotency; Ulk1; p53; p53(−/−)
    DOI:  https://doi.org/10.1016/j.bbrc.2021.03.034
  51. FEBS J. 2021 Mar 22.
    ATG16L1 functions in cell homeostasis beyond autophagy.
    Daniel Hamaoui, Agathe Subtil.
      Atg16-like (ATG16L) proteins were identified in higher eukaryotes for their resemblance to Atg16, a yeast protein previously characterized as a subunit of the Atg12-Atg5/Atg16 complex. In yeast, this complex catalyzes the lipidation of Atg8 on pre-autophagosomal structures, and is thereby required for the formation of autophagosomes. In higher eukaryotes, ATG16L1 is also almost exclusively present as part of an ATG12-ATG5/ATG16L1 complex, and has the same essential function in autophagy. However, ATG16L1 is three times bigger than Atg16. It displays in particular a carboxy-terminal extension, including a WD40 domain, which provides a platform for interaction with a variety of proteins, and allows for the recruitment of the ATG12-ATG5/ATG16L1 complex to membranes under different contexts. Furthermore, detailed analyses at the cellular level have revealed that some of the ATG16L1-driven activities are independent of the lipidation reaction catalyzed by the ATG12-ATG5/ATG16L1 complex. At the organ level, the use of mice that are hypomorphic for Atg16l1, or with cell-specific ablation of its expression, revealed a large panel of consequences of ATG16L1 dysfunctions. In this review, we recapitulate the current knowledge on ATG16L1 expression and functions. We emphasize in particular how it broadly acts as a brake on inflammation, thereby contributing to maintaining cell homeostasis. We also report on independent studies that converge to show that ATG16L1 is an important player in the regulation of intracellular traffic. Overall, autophagy-independent functions of ATG16L1 probably accounts for more of the phenotypes associated with ATG16L1 deficiencies than currently appreciated.
    Keywords:  ATG5-ATG12/ATG16L1 complex; Crohn’s disease; LC3-associated phagocytosis; LC3-lipidation; T300A variant; WD40 domain; autophagy
    DOI:  https://doi.org/10.1111/febs.15833
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