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Bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle

Aging is associated with increasing incidence of osteoporosis; a skeletal disorder characterized by compromised bone strength that may predispose patients to an increased risk of fracture. It is imperative to identify novel ways in which to attenuate such declines in the functional properties of bon...

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Autores principales: Forrester, Steven J., Kawata, Keisuke, Lee, Hojun, Kim, Ji‐Seok, Sebzda, Kelly, Butler, Tiffiny, Yingling, Vanessa R., Park, Joon‐Young
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Wiley Periodicals, Inc. 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4332228/
https://www.ncbi.nlm.nih.gov/pubmed/25539834
http://dx.doi.org/10.14814/phy2.12255
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author Forrester, Steven J.
Kawata, Keisuke
Lee, Hojun
Kim, Ji‐Seok
Sebzda, Kelly
Butler, Tiffiny
Yingling, Vanessa R.
Park, Joon‐Young
author_facet Forrester, Steven J.
Kawata, Keisuke
Lee, Hojun
Kim, Ji‐Seok
Sebzda, Kelly
Butler, Tiffiny
Yingling, Vanessa R.
Park, Joon‐Young
author_sort Forrester, Steven J.
collection PubMed
description Aging is associated with increasing incidence of osteoporosis; a skeletal disorder characterized by compromised bone strength that may predispose patients to an increased risk of fracture. It is imperative to identify novel ways in which to attenuate such declines in the functional properties of bone. The purpose of this study was to identify, through in silico, in vitro, and in vivo approaches, a protein secreted from skeletal muscle that is putatively involved in bone formation. We performed a functional annotation bioinformatic analysis of human skeletal muscle‐derived secretomes (n = 319) using DAVID software. Cross‐referencing was conducted using OMIM, Unigene, UniProt, GEO, and CGAP databases. Signal peptides and transmembrane residues were analyzed using SignalP and TMHMM software. To further investigate functionality of the identified protein, L6 and C2C12 myotubes were grown for in vitro analysis. C2C12 myotubes were subjected to 16 h of glucose deprivation (GD) prior to analysis. In vivo experiments included analysis of 6‐week calorie restricted (CR) rat muscle samples. Bioinformatic analysis yielded 15 genes of interest. GEO dataset analysis identified BMP5, COL1A2, CTGF, MGP, MMP2, and SPARC as potential targets for further processing. Following TMHMM and SignalP processing, CTGF was chosen as a candidate gene. CTGF expression level was increased during L6 myoblast differentiation (P <0.01). C2C12 myotubes showed no change in response to GD. Rat soleus muscle samples exhibited an increase in CTGF expression (n = 16) in response to CR (35%) (P <0.05). CTGF was identified as a skeletal muscle expressed protein through bioinformatic analysis of skeletal muscle‐derived secretomes and in vitro/in vivo analysis. Future study is needed to determine the role of muscle‐derived CTGF in bone formation and remodeling processes.
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spelling pubmed-43322282015-04-07 Bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle Forrester, Steven J. Kawata, Keisuke Lee, Hojun Kim, Ji‐Seok Sebzda, Kelly Butler, Tiffiny Yingling, Vanessa R. Park, Joon‐Young Physiol Rep Original Research Aging is associated with increasing incidence of osteoporosis; a skeletal disorder characterized by compromised bone strength that may predispose patients to an increased risk of fracture. It is imperative to identify novel ways in which to attenuate such declines in the functional properties of bone. The purpose of this study was to identify, through in silico, in vitro, and in vivo approaches, a protein secreted from skeletal muscle that is putatively involved in bone formation. We performed a functional annotation bioinformatic analysis of human skeletal muscle‐derived secretomes (n = 319) using DAVID software. Cross‐referencing was conducted using OMIM, Unigene, UniProt, GEO, and CGAP databases. Signal peptides and transmembrane residues were analyzed using SignalP and TMHMM software. To further investigate functionality of the identified protein, L6 and C2C12 myotubes were grown for in vitro analysis. C2C12 myotubes were subjected to 16 h of glucose deprivation (GD) prior to analysis. In vivo experiments included analysis of 6‐week calorie restricted (CR) rat muscle samples. Bioinformatic analysis yielded 15 genes of interest. GEO dataset analysis identified BMP5, COL1A2, CTGF, MGP, MMP2, and SPARC as potential targets for further processing. Following TMHMM and SignalP processing, CTGF was chosen as a candidate gene. CTGF expression level was increased during L6 myoblast differentiation (P <0.01). C2C12 myotubes showed no change in response to GD. Rat soleus muscle samples exhibited an increase in CTGF expression (n = 16) in response to CR (35%) (P <0.05). CTGF was identified as a skeletal muscle expressed protein through bioinformatic analysis of skeletal muscle‐derived secretomes and in vitro/in vivo analysis. Future study is needed to determine the role of muscle‐derived CTGF in bone formation and remodeling processes. Wiley Periodicals, Inc. 2014-12-24 /pmc/articles/PMC4332228/ /pubmed/25539834 http://dx.doi.org/10.14814/phy2.12255 Text en © 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Research
Forrester, Steven J.
Kawata, Keisuke
Lee, Hojun
Kim, Ji‐Seok
Sebzda, Kelly
Butler, Tiffiny
Yingling, Vanessa R.
Park, Joon‐Young
Bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle
title Bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle
title_full Bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle
title_fullStr Bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle
title_full_unstemmed Bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle
title_short Bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle
title_sort bioinformatic identification of connective tissue growth factor as an osteogenic protein within skeletal muscle
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4332228/
https://www.ncbi.nlm.nih.gov/pubmed/25539834
http://dx.doi.org/10.14814/phy2.12255
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