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Functional genomic screen identifies novel mediators of collagen uptake

Tissue fibrosis occurs when matrix production outpaces matrix degradation. Degradation of collagen, the main component of fibrotic tissue, is mediated through an extracellular proteolytic pathway and intracellular pathway of cellular uptake and lysosomal digestion. Recent studies demonstrate that di...

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Autores principales: Lee, Ting-Hein, McKleroy, William, Khalifeh-Soltani, Amin, Sakuma, Stephen, Lazarev, Stanislav, Riento, Kirsi, Nishimura, Stephen L., Nichols, Ben J., Atabai, Kamran
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3937085/
https://www.ncbi.nlm.nih.gov/pubmed/24403604
http://dx.doi.org/10.1091/mbc.E13-07-0382
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author Lee, Ting-Hein
McKleroy, William
Khalifeh-Soltani, Amin
Sakuma, Stephen
Lazarev, Stanislav
Riento, Kirsi
Nishimura, Stephen L.
Nichols, Ben J.
Atabai, Kamran
author_facet Lee, Ting-Hein
McKleroy, William
Khalifeh-Soltani, Amin
Sakuma, Stephen
Lazarev, Stanislav
Riento, Kirsi
Nishimura, Stephen L.
Nichols, Ben J.
Atabai, Kamran
author_sort Lee, Ting-Hein
collection PubMed
description Tissue fibrosis occurs when matrix production outpaces matrix degradation. Degradation of collagen, the main component of fibrotic tissue, is mediated through an extracellular proteolytic pathway and intracellular pathway of cellular uptake and lysosomal digestion. Recent studies demonstrate that disruption of the intracellular pathways can exacerbate fibrosis. These pathways are poorly characterized. Here we identify novel mediators of the intracellular pathway of collagen turnover through a genome-wide RNA interference screen in Drosophila S2 cells. Screening of 7505 Drosophila genes conserved among metazoans identified 22 genes that were required for efficient internalization of type I collagen. These included proteins involved in vesicle transport, the actin cytoskeleton, and signal transduction. We show further that the flotillin genes have a conserved and central role in collagen uptake in Drosophila and human cells. Short hairpin RNA–mediated silencing of flotillins in human monocyte and fibroblasts impaired collagen uptake by promoting lysosomal degradation of the endocytic collagen receptors uPARAP/Endo180 and mannose receptor. These data provide an initial characterization of intracellular pathways of collagen turnover and identify the flotillin genes as critical regulators of this process. A better understanding of these pathways may lead to novel therapies that reduce fibrosis by increasing collagen turnover.
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spelling pubmed-39370852014-05-16 Functional genomic screen identifies novel mediators of collagen uptake Lee, Ting-Hein McKleroy, William Khalifeh-Soltani, Amin Sakuma, Stephen Lazarev, Stanislav Riento, Kirsi Nishimura, Stephen L. Nichols, Ben J. Atabai, Kamran Mol Biol Cell Articles Tissue fibrosis occurs when matrix production outpaces matrix degradation. Degradation of collagen, the main component of fibrotic tissue, is mediated through an extracellular proteolytic pathway and intracellular pathway of cellular uptake and lysosomal digestion. Recent studies demonstrate that disruption of the intracellular pathways can exacerbate fibrosis. These pathways are poorly characterized. Here we identify novel mediators of the intracellular pathway of collagen turnover through a genome-wide RNA interference screen in Drosophila S2 cells. Screening of 7505 Drosophila genes conserved among metazoans identified 22 genes that were required for efficient internalization of type I collagen. These included proteins involved in vesicle transport, the actin cytoskeleton, and signal transduction. We show further that the flotillin genes have a conserved and central role in collagen uptake in Drosophila and human cells. Short hairpin RNA–mediated silencing of flotillins in human monocyte and fibroblasts impaired collagen uptake by promoting lysosomal degradation of the endocytic collagen receptors uPARAP/Endo180 and mannose receptor. These data provide an initial characterization of intracellular pathways of collagen turnover and identify the flotillin genes as critical regulators of this process. A better understanding of these pathways may lead to novel therapies that reduce fibrosis by increasing collagen turnover. The American Society for Cell Biology 2014-03-01 /pmc/articles/PMC3937085/ /pubmed/24403604 http://dx.doi.org/10.1091/mbc.E13-07-0382 Text en © 2014 Lee et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
Lee, Ting-Hein
McKleroy, William
Khalifeh-Soltani, Amin
Sakuma, Stephen
Lazarev, Stanislav
Riento, Kirsi
Nishimura, Stephen L.
Nichols, Ben J.
Atabai, Kamran
Functional genomic screen identifies novel mediators of collagen uptake
title Functional genomic screen identifies novel mediators of collagen uptake
title_full Functional genomic screen identifies novel mediators of collagen uptake
title_fullStr Functional genomic screen identifies novel mediators of collagen uptake
title_full_unstemmed Functional genomic screen identifies novel mediators of collagen uptake
title_short Functional genomic screen identifies novel mediators of collagen uptake
title_sort functional genomic screen identifies novel mediators of collagen uptake
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3937085/
https://www.ncbi.nlm.nih.gov/pubmed/24403604
http://dx.doi.org/10.1091/mbc.E13-07-0382
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