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Role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis
OBJECTIVES: To elucidate mechanisms contributing to skeletal muscle calcinosis in patients with juvenile dermatomyositis. METHODS: A well-characterized cohorts of JDM (n = 68), disease controls (polymyositis, n = 7; juvenile SLE, n = 10, and RNP + overlap syndrome, n = 12), and age-matched health co...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10330803/ https://www.ncbi.nlm.nih.gov/pubmed/37244073 http://dx.doi.org/10.1016/j.jaut.2023.103061 |
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author | Duvvuri, Bhargavi Pachman, Lauren M. Hermanson, Payton Wang, Ting Moore, Richard Wang, Dennis Ding-Hwa Long, Aaron Morgan, Gabrielle A. Doty, Stephen Tian, Rong Sancak, Yasemin Lood, Christian |
author_facet | Duvvuri, Bhargavi Pachman, Lauren M. Hermanson, Payton Wang, Ting Moore, Richard Wang, Dennis Ding-Hwa Long, Aaron Morgan, Gabrielle A. Doty, Stephen Tian, Rong Sancak, Yasemin Lood, Christian |
author_sort | Duvvuri, Bhargavi |
collection | PubMed |
description | OBJECTIVES: To elucidate mechanisms contributing to skeletal muscle calcinosis in patients with juvenile dermatomyositis. METHODS: A well-characterized cohorts of JDM (n = 68), disease controls (polymyositis, n = 7; juvenile SLE, n = 10, and RNP + overlap syndrome, n = 12), and age-matched health controls (n = 17) were analyzed for circulating levels of mitochondrial (mt) markers including mtDNA, mt-nd6, and anti-mitochondrial antibodies (AMAs) using standard qPCR, ELISA, and novel-in-house assays, respectively. Mitochondrial calcification of affected tissue biopsies was confirmed using electron microscopy and energy dispersive X-ray analysis. A human skeletal muscle cell line, RH30, was used to generate an in vitro calcification model. Intracellular calcification is measured by flow cytometry and microscopy. Mitochondria were assessed for mtROS production and membrane potential by flow cytometry and real-time oxygen consumption rate by Seahorse bioanalyzer. Inflammation (interferon-stimulated genes) was measured by qPCR. RESULTS: In the current study, patients with JDM exhibited elevated levels of mitochondrial markers associated with muscle damage and calcinosis. Of particular interest are AMAs predictive of calcinosis. Human skeletal muscle cells undergo time- and dose-dependent accumulation of calcium phosphate salts with preferential localization to mitochondria. Calcification renders skeletal muscle cells mitochondria stressed, dysfunctional, destabilized, and interferogenic. Further, we report that inflammation induced by interferon-alpha amplifies mitochondrial calcification of human skeletal muscle cells via the generation of mitochondrial reactive oxygen species (mtROS). CONCLUSIONS: Overall, our study demonstrates the mitochondrial involvement in the skeletal muscle pathology and calcinosis of JDM and mtROS as a central player in the calcification of human skeletal muscle cells. Therapeutic targeting of mtROS and/or upstream inducers, such as inflammation, may alleviate mitochondrial dysfunction, leading to calcinosis. AMAs can potentially identify patients with JDM at risk for developing calcinosis. |
format | Online Article Text |
id | pubmed-10330803 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
record_format | MEDLINE/PubMed |
spelling | pubmed-103308032023-07-10 Role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis Duvvuri, Bhargavi Pachman, Lauren M. Hermanson, Payton Wang, Ting Moore, Richard Wang, Dennis Ding-Hwa Long, Aaron Morgan, Gabrielle A. Doty, Stephen Tian, Rong Sancak, Yasemin Lood, Christian J Autoimmun Article OBJECTIVES: To elucidate mechanisms contributing to skeletal muscle calcinosis in patients with juvenile dermatomyositis. METHODS: A well-characterized cohorts of JDM (n = 68), disease controls (polymyositis, n = 7; juvenile SLE, n = 10, and RNP + overlap syndrome, n = 12), and age-matched health controls (n = 17) were analyzed for circulating levels of mitochondrial (mt) markers including mtDNA, mt-nd6, and anti-mitochondrial antibodies (AMAs) using standard qPCR, ELISA, and novel-in-house assays, respectively. Mitochondrial calcification of affected tissue biopsies was confirmed using electron microscopy and energy dispersive X-ray analysis. A human skeletal muscle cell line, RH30, was used to generate an in vitro calcification model. Intracellular calcification is measured by flow cytometry and microscopy. Mitochondria were assessed for mtROS production and membrane potential by flow cytometry and real-time oxygen consumption rate by Seahorse bioanalyzer. Inflammation (interferon-stimulated genes) was measured by qPCR. RESULTS: In the current study, patients with JDM exhibited elevated levels of mitochondrial markers associated with muscle damage and calcinosis. Of particular interest are AMAs predictive of calcinosis. Human skeletal muscle cells undergo time- and dose-dependent accumulation of calcium phosphate salts with preferential localization to mitochondria. Calcification renders skeletal muscle cells mitochondria stressed, dysfunctional, destabilized, and interferogenic. Further, we report that inflammation induced by interferon-alpha amplifies mitochondrial calcification of human skeletal muscle cells via the generation of mitochondrial reactive oxygen species (mtROS). CONCLUSIONS: Overall, our study demonstrates the mitochondrial involvement in the skeletal muscle pathology and calcinosis of JDM and mtROS as a central player in the calcification of human skeletal muscle cells. Therapeutic targeting of mtROS and/or upstream inducers, such as inflammation, may alleviate mitochondrial dysfunction, leading to calcinosis. AMAs can potentially identify patients with JDM at risk for developing calcinosis. 2023-07 2023-05-25 /pmc/articles/PMC10330803/ /pubmed/37244073 http://dx.doi.org/10.1016/j.jaut.2023.103061 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) ). |
spellingShingle | Article Duvvuri, Bhargavi Pachman, Lauren M. Hermanson, Payton Wang, Ting Moore, Richard Wang, Dennis Ding-Hwa Long, Aaron Morgan, Gabrielle A. Doty, Stephen Tian, Rong Sancak, Yasemin Lood, Christian Role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis |
title | Role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis |
title_full | Role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis |
title_fullStr | Role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis |
title_full_unstemmed | Role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis |
title_short | Role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis |
title_sort | role of mitochondria in the myopathy of juvenile dermatomyositis and implications for skeletal muscle calcinosis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10330803/ https://www.ncbi.nlm.nih.gov/pubmed/37244073 http://dx.doi.org/10.1016/j.jaut.2023.103061 |
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