Diabetes diminishes muscle precursor cell-mediated microvascular angiogenesis

PLOS ONE RESEARCH ARTICLE Diabetes diminishes muscle precursor cellmediated microvascular angiogenesis Francisca M. Acosta ID1,2¤a, Settimio Pacelli1¤b, Christopher R. Rathbone ID1,2,3* 1 Department of Biomedical and Chemical Engineering, University of Texas at San Antonio, San Antonio, TX, United States of America, 2 UTSA-UTHSCSA Joint Graduate Program in Biomedical Engineering, San Antonio, TX, United States of America, 3 Institute of Regenerative Medicine, University of Texas at San Antonio, San Antonio, TX, United States of America a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 ¤a Current address: Department of Biochemistry and Structural Biology, University of Texas Health Science Center, San Antonio, TX, United States of America ¤b Current address: Department of Biomedical Engineering, Illinois Institute of Technology, Chicago, IL, United States of America * Abstract OPEN ACCESS Citation: Acosta FM, Pacelli S, Rathbone CR (2023) Diabetes diminishes muscle precursor cellmediated microvascular angiogenesis. PLoS ONE 18(8): e0289477. https://doi.org/10.1371/journal. pone.0289477 Editor: Kanhaiya Singh, Indiana University Purdue University at Indianapolis, UNITED STATES Received: January 26, 2023 Accepted: July 19, 2023 Published: August 4, 2023 Copyright: © 2023 Acosta et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: This work was financially supported by the National Institutes of Health in the form of a grant (SC1DK122578) awarded to CRR. This work was also financially supported by the National Institutes of Health in the form of trainee support awarded to FMA through an institutional grant (GM060655). The Bioanalytics and Single-Cell Core at UTHSCSA, which is supported by CPRIT grant (RP150600) and the Office of Vice President of The skeletal muscles of Type II diabetic (T2D) patients can be characterized by a reduced vessel density, corresponding to deficiencies in microvascular angiogenesis. Interestingly, T2D also inhibits the function of many myogenic cells resident within skeletal muscle, including satellite cells, which are well-known for the role they play in maintaining homeostasis. The current study was undertaken to gain a better understanding of the mechanisms whereby satellite cell progeny, muscle precursor cells (MPCs), influence microvascular angiogenesis. Network growth and the expression of genes associated with angiogenesis were reduced when microvessels were treated with conditioned media generated by proliferating MPCs isolated from diabetic, as compared to control rat skeletal muscle, a phenomenon that was also observed when myoblasts from control or diabetic human skeletal muscle were used. When only exosomes derived from diabetic or control MPCs were used to treat microvessels, no differences in microvascular growth were observed. An evaluation of the angiogenesis factors in control and diabetic MPCs revealed differences in Leptin, vascular endothelial growth factor (VEGF), IL1-β, interleukin 10, and IP-10, and an evaluation of the MPC secretome revealed differences in interleukin 6, MCP-1, VEGF, and interleukin 4 exist. Angiogenesis was also reduced in tissue-engineered skeletal muscles (TE-SkM) containing microvessels when they were generated from MPCs isolated from diabetic as compared to control skeletal muscle. Lastly, the secretome of injured control, but not diabetic, TE-SkM was able to increase VEGF and increase microvascular angiogenesis. This comprehensive analysis of the interaction between MPCs and microvessels in the context of diabetes points to an area for alleviating the deleterious effects of diabetes on skeletal muscle. PLOS ONE | https://doi.org/10.1371/journal.pone.0289477 August 4, 2023 1 / 20 PLOS ONE Research at UT Health SA, was utilized for the completion of these studies. The funders had no additional role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist. Diabetes diminishes muscle precursor cell-mediated microvascular angiogenesis Introduction Given the importance of the perfusion of skeletal muscle fibers in maintaining homeostasis, it is intuitive that reductions in capillary density and disruptions in the microvascular function that accompany diseases, like Type II diabetes (T2D), are associated with deleterious consequences in skeletal muscle [1–3]. Satellite cells, resident adult muscle stem cells, are widely recognized for their contribution to muscle homeostasis and muscle repair through their direct participation in myogenesis. The ability of satellite cells to influence their microenvironment through the secretion of factors that impact, for example, neurogenesis, inflammation, and angiogenesis, to name a few, is recognized as an additional satellite cell attribute [4–6]. With regards to the latter, the observation that resident stem cells within skeletal muscle called satellite cells are closely associated with blood vessels in vivo, and that the number of satellite cells positively correlates with capillarization implies that an association between satellite cells and microvessels may contribute to muscle homeostasis by affecting the vasculature [7, 8]. Collectively, when specifically taking T2D into consideration, the knowledge that satellite cell content, regenerative capacity, and capillary density are all reduced in muscles in diabetes and/or obesity suggests the interaction between satellite cells and microvessels in diabetic muscle is a critical facet of the disease [9–14]. In vitro experiments support the idea that satellite cells exert a positive influence over microvessels. Factors secreted by the progeny of satellite cells (myoblasts, MPCs, MuSCs, etc.) or their immortalized cell equivalent (C2C12 cells) can stimulate angiogenesis [15, 16]. For the remainder of the manuscript, the progeny of satellite cells will be referred to as muscle precursor cells (MPCs) where appropriate. Following in line with the idea that this interaction may be impaired in the context of disease, secretions from muscle cells taken from unhealthy tissue, or treated to resemble diseased cells, have a reduced ability to stimulate angiogenesis [17–20]. The types and/or quantity of growth factors secreted by diseased satellite cells (or their in vitro progeny) that are responsible for this insufficiency are beginning to be delineated. For example, myotubes derived from T2D subjects produced an increase in the level of Interleukin 8 that was associated with a reduction in human umbilical vein (HUVEC) tube formation and capillary outgrowth [19]. Conversely, decrea (...truncated)