Repertoires of Autophagy in the Pathogenesis of Ocular Diseases

Cellular Physiology and Biochemistry, Journal Year: 2015, Volume and Issue: 35(5), P. 1663 - 1676

Published: Jan. 1, 2015

Age-related macular degeneration (AMD) is the most common reason of visual impairment in elderly Western countries. The retinal pigment epithelial cells (RPE) causes secondarily adverse effects on neural retina leading to loss. aging characteristics RPE involve lysosomal accumulation lipofuscin and extracellular protein aggregates called "drusen". Molecular mechanisms behind aggregations are weakly understood. There intriguing evidence suggesting that SQSTM1/p62, together with autophagy, has a role pathology different degenerative diseases. It appears SQSTM1/p62 connecting link between autophagy proteasome mediated proteolysis, expressed strongly under exposure various oxidative stimuli proteasomal inhibition. ELAVL1/HuR post-transcriptional factor, which acts mainly as positive regulator gene expression by binding specific mRNAs whose corresponding proteins fundamental for key cellular functions. We here show that, inhibitor MG-132, up-regulated at both mRNA levels, this binds post-transcriptionally regulates ARPE-19 cell line. Furthermore, we observed inhibition caused bound irreversibly perinuclear aggregates. addition AMPK activator AICAR was pro-survival promoted cleansing former complex, but not accumulation, indeed decreased through autophagy-mediated degradation, while pathway. Interestingly, when compared human controls, AMD donor samples strong rather than drusen rich area impaired AMD.

Language: Английский

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Understanding the Warburg Effect: The Metabolic Requirements of Cell Proliferation

Science, Journal Year: 2009, Volume and Issue: 324(5930), P. 1029 - 1033

Published: May 21, 2009

In contrast to normal differentiated cells, which rely primarily on mitochondrial oxidative phosphorylation generate the energy needed for cellular processes, most cancer cells instead aerobic glycolysis, a phenomenon termed "the Warburg effect." Aerobic glycolysis is an inefficient way adenosine 5'-triphosphate (ATP), however, and advantage it confers has been unclear. Here we propose that metabolism of indeed all proliferating adapted facilitate uptake incorporation nutrients into biomass (e.g., nucleotides, amino acids, lipids) produce new cell. Supporting this idea are recent studies showing (i) several signaling pathways implicated in cell proliferation also regulate metabolic incorporate biomass; (ii) certain cancer-associated mutations enable acquire metabolize manner conducive rather than efficient ATP production. A better understanding mechanistic links between growth control may ultimately lead treatments human cancer.

Language: Английский

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The Hallmarks of Aging

Cell, Journal Year: 2013, Volume and Issue: 153(6), P. 1194 - 1217

Published: June 1, 2013

Language: Английский

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mTOR Signaling in Growth, Metabolism, and Disease

Cell, Journal Year: 2017, Volume and Issue: 168(6), P. 960 - 976

Published: March 1, 2017

Language: Английский

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Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Autophagy, Journal Year: 2016, Volume and Issue: 12(1), P. 1 - 222

Published: Jan. 2, 2016

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, on this topic has continued to accelerate, and many new scientists have entered field. Our knowledge base relevant technologies also been expanding. Accordingly, it is important update these monitoring autophagy different organisms. Various reviews described range assays that used purpose. Nevertheless, there continues be confusion regarding acceptable methods measure autophagy, especially multicellular eukaryotes. For example, a key point needs emphasized difference between measurements monitor numbers or volume autophagic elements (e.g., autophagosomes autolysosomes) at any stage process versus those flux through pathway (i.e., complete including amount rate cargo sequestered degraded). particular, block macroautophagy results autophagosome accumulation must differentiated from stimuli increase activity, defined as increased induction coupled with delivery to, degradation within, lysosomes (in most higher eukaryotes some protists such Dictyostelium) vacuole plants fungi). other words, investigators field understand appearance more does not necessarily equate fact, cases, accumulate because trafficking without concomitant change biogenesis, whereas an autolysosomes may reflect reduction degradative activity. It worth emphasizing here lysosomal digestion evaluating its competence crucial part evaluation flux, autophagy. Here, present selection interpretation use by who aim examine related processes, well reviewers need provide realistic reasonable critiques papers are focused processes. These meant formulaic rules, appropriate depend question being asked system used. addition, emphasize no individual assay guaranteed one every situation, strongly recommend multiple Along lines, potential pleiotropic effects due blocking genetic manipulation, imperative target gene knockout RNA interference than autophagy-related protein. Atg proteins, groups involved cellular pathways implying all proteins can specific marker process. guidelines, consider various assessing what information can, cannot, obtained them. Finally, discussing merits limits particular assays, hope encourage technical innovation

Language: Английский

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Anti-Vascular Endothelial Growth Factors Protect Retinal Pigment Epithelium Cells Against Oxidation by Modulating Nitric Oxide Release and Autophagy

Cellular Physiology and Biochemistry, Journal Year: 2017, Volume and Issue: 42(5), P. 1725 - 1738

Published: Jan. 1, 2017

To investigate whether oxidative stress modulates vascular endothelial growth factor (VEGF)-A and VEGF-C expression polarized secretion in a human retinal pigment epithelium cell line (ARPE-19).Long-term culture of ARPE-19 cells Dulbecco's modified Eagle medium (DMEM)/F12 containing 1% fetal bovine serum (FBS) on transwell filters (12 mm or 6 mm, pore size 0.4 microm) was performed to produce (RPE) monolayers. The integrity monolayer established by measurement transepithelial resistance (TER) presence tight junctions assessed zonula occludens (ZO-1) occludin apical Na/K ATPase localization. Paracellular permeability studied using radiolabeled mannitol. Confluent were treated with tertiary butyl hydrogen peroxide (tBH) for varying durations (0-5 h) doses (50-200 microM). VEGF-A -C evaluated western blot quantitative RT-PCR, while the basolateral surfaces quantitated ELISA.Polarity verified localization junction proteins, ZO-1 its binding partner confocal microscopy as well Na,K-ATPase at surface. TER confluent averaged 48.7+/-2.1 Omega. cm(2) tBH treatment did not alter significantly (46.9+/-1.9 cm(2); p>0.05 versus controls) expression. Whole mRNA nonpolarized increased 5 h both increase significant (p<0.05 vs controls). A similar, maximal also observed cellular protein levels. ARPE showed an exposure. levels higher monolayers stimulated domains. 150 microM function time (1-5 increases from 410 2080 pg/10(6) 290 1680 pattern similar. dose-dependent range 50-200 tended be than secretion.Our data show that RPE upregulates C. side Given role VEGF choroidal neovascularization, these may value understanding pathogenic mechanisms designing antiangiogenic therapies.

Language: Английский

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The Emerging Hallmarks of Cancer Metabolism

Cell Metabolism, Journal Year: 2016, Volume and Issue: 23(1), P. 27 - 47

Published: Jan. 1, 2016

Language: Английский

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Autophagy and Viruses: Adversaries or Allies?

Journal of Innate Immunity, Journal Year: 2013, Volume and Issue: 5(5), P. 480 - 493

Published: Jan. 1, 2013

Hepatitis C virus (HCV) is a major cause of chronic liver diseases. A high risk chronicity the concern HCV infection, since infection often leads to cirrhosis and hepatocellular carcinoma. Infection with genotype 1 in particular considered clinical factor for development carcinoma, although molecular mechanisms pathogenesis are largely unknown. Autophagy involved degradation cellular organelles elimination invasive microorganisms. In addition, disruption autophagy several protein deposition Although recent reports suggest that exploits pathway viral propagation, biological significance life cycle still uncertain. Here, we show replication RNA induces inhibit cell death. Cells harboring an replicon 1b strain Con1 but not 2a JFH1 exhibited incomplete acidification autolysosome due lysosomal defect, leading enhanced secretion immature cathepsin B. The suppression cells induced severe cytoplasmic vacuolation These results harnesses circumvent harmful vacuole formation maintain persistent infection. findings reveal unique survival strategy provide new insights into genotype-specific pathogenicity HCV.

Language: Английский

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Guidelines for the use and interpretation of assays for monitoring autophagy

Autophagy, Journal Year: 2012, Volume and Issue: 8(4), P. 445 - 544

Published: April 1, 2012

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, on this topic has continued to accelerate, and many new scientists have entered field. Our knowledge base relevant technologies also been expanding. Accordingly, it is important update these monitoring autophagy different organisms. Various reviews described range assays that used purpose. Nevertheless, there continues be confusion regarding acceptable methods measure autophagy, especially multicellular eukaryotes. A key point needs emphasized a difference between measurements monitor numbers or volume autophagic elements (e.g., autophagosomes autolysosomes) at any stage process vs. those flux through pathway (i.e., complete process); thus, block macroautophagy results autophagosome accumulation differentiated from stimuli result increased activity, defined as induction coupled with delivery to, degradation within, lysosomes (in most higher eukaryotes some protists such Dictyostelium) vacuole plants fungi). other words, investigators field understand appearance more does not necessarily equate fact, cases, accumulate because trafficking without concomitant change biogenesis, whereas an increase autolysosomes may reflect reduction degradative activity. Here, present selection interpretation use by who aim examine related processes, well reviewers need provide realistic reasonable critiques papers are focused processes. These meant formulaic rules, appropriate depend part question being asked system used. addition, emphasize no individual assay guaranteed one every situation, strongly recommend multiple guidelines, consider various assessing what information can, cannot, obtained them. Finally, discussing merits limits particular assays, hope encourage technical innovation

Language: Английский

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Antimicrobial Autophagy: A Conserved Innate Immune Response in Drosophila

Journal of Innate Immunity, Journal Year: 2013, Volume and Issue: 5(5), P. 444 - 455

Published: Jan. 1, 2013

Autophagy is a major route by which cytoplasmic contents are delivered to the lysosome for degradation. Many autophagy-related (ATG) genes have been identified in yeast. Although most of them conserved human, molecular composition Atg1 complex appears differ between yeast and mammals. In yeast, forms with Atg11, Atg13, Atg17, Atg29 Atg31, whereas mammalian (ULK1/2) interacts Atg13 FIP200. Here, we identify novel binding protein, named Atg101. Atg101 shows no homology other Atg proteins, various eukaryotes, but not Saccharomyces cerevisiae. associates ULK-Atg13-FIP200 complex, likely through direct interaction Atg13. siRNA-treated cells, present solely as monomer. Interaction stable, regulated nutrient conditions. GFP-Atg101 localizes isolation membrane/phagophore. GFP-LC3 dot formation suppressed endogenous LC3-I accumulates suggesting that critical factor autophagy. Furthermore, important stability basal phosphorylation ULK1. These data suggest protein functions together ULK,

Language: Английский

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