Exosomal‐ribosomal proteins‐driven heterogeneity of epicardial adipose tissue derived stem cells under ischemia for cardiac regeneration

Abstract

Extracellular ribosomal proteins secreted in exosomes elicit biological/regenerative responses; however, ribosomal proteins contained in the exosomes of ischemiachallenged epicardial adipose tissue-derived stem cells (EATDS) remain unexplored. This study focuses on the identification of ribosomal proteins in the exosomes of ischemia-challenged EATDS and their sub-populations based on the key ribosomal proteins using single-cell genomics. Exosomes were isolated from control, ischemic (ISC), and reperfused (ISC/R) EATDS harvested from hyperlipidemic microswine, and the proteins were detected using Liquid chromatography with tandem mass spectrometry (LC-MS/MS). One hundred ninety-nine proteins and 177 proteins were detected in ISC and ISC/R groups, respectively with significant fold-change compared to controls. Five ribosomal proteins, RPL10A, 40SRPS18, 40SRPS30, 60SRPL14, and 40SRPSA, were significant owing to their abundance based on LC-MS/MS data. Expression of these proteins, except RPL10A, at transcript and protein levels were lower in ISC group compared to the control. scRNAseq analysis revealed EATDS heterogeneity based on the upregulation of 40SRPSA, 40SRPL18, and 40SRPS18. Pro-inflammatory sub-populations upregulated CCL5, anti-inflammatory sub-population upregulated IL-11, proliferative sub-population upregulated cell cycle and DNA replication mediators, and nonproliferative population downregulated the cell cycle and DNA replication mediators. Overall, the functional role of extracellular ribosomal proteins in driving unique phenotypes of EATDS population offers promise for designing effective translational approaches for myocardial regeneration.

Unbiased proteomic analysis of extracellular vesicles secreted by senescent human vascular smooth muscle cells reveals their ability to modulate immune cell functions

“…For instance EVs secreted by LPS-mediated microglia were enriched in ribosomal proteins [ 26 ]. Different ribosomal proteins were also identified in the exosomes of ischemia challenged epicardial adipose tissue–derived stem cells [ 27 ]. The biological meaning of those proteins present in EVs is still unrevealed.…”

Carboxymethyl cellulose–alginate interpenetrating hydroxy ethyl methacrylate crosslinked polyvinyl alcohol reinforced hybrid hydrogel templates with improved biological performance for cardiac tissue engineering

“…CAHA2A and CAHA2AP hydrogels were imbibed in distilled water to determine swelling kinetics, swelling ratio ( S ), equilibrium water content (EWC), and equilibrium swelling ratio (ESR) following our previous protocols (Thankam, Diaz, et al, 2022; Thankam, Huynh, et al, 2022). $$<math altimg="urn:x-wiley:00063592:media:bit28291:bit28291-math-0001" display="block" wiley:location="equation/bit28291-math-0001.png" xmlns="http://www.w3.org/1998/Math/MathML"><mrow><mrow><mi>Swelling</mi><mspace width="0.25em"/><mi>ratio</mi><mspace width="0.25em"/><mrow><mo>(</mo><mi>S</mi><mo>)</mo></mrow><mo>\unicode{x0003D}</mo><mspace/><mfrac><mrow><msub><mi>M</mi><mi mathvariant="normal">t</mi></msub><mo>\unicode{x02212}</mo><msub><mi>M</mi><mn>0</mn></msub></mrow><msub><mi>M</mi><mn>0</mn></msub></mfrac><mo>,</mo></mrow></mrow></math>$$ $$<math altimg="urn:x-wiley:00063592:media:bit28291:bit28291-math-0002" display="block" wiley:location="equation/bit28291-math-0002.png" xmlns="http://www.w3.org/1998/Math/MathML"><mrow><mrow><mo>$$ $$<math altimg="urn:x-wiley:00063592:media:bit28291:bit28291-math-0003" display="block" wiley:location="equation/bit28291-math-0003.png" xmlns="http://www.w3.org/1998/Math/MathML"><mrow><mrow><mi>ESR</mi><mo>(</mo><mi>E</mi><mo>)</mo><mo>\unicode{x0003D}</mo><mspace/><mfrac><mrow><msub><mi>M</mi><mi mathvar...$$…”

“…CAHA2A and CAHA2AP hydrogels were imbibed in distilled water to determine swelling kinetics, swelling ratio (S), equilibrium water content (EWC), and equilibrium swelling ratio (ESR) following our previous protocols (Thankam, Diaz, et al, 2022;Thankam, Huynh, et al, 2022).…”

“…The EATDS and CF cells were harvested and characterized based on the immunopositivity of specific biomarkers (Thankam, Huynh, et al, 2022) (data not shown in this article) and were labeled with viable fluorescent dyes displaying green and red fluorescence. Both the hydrogels supported the coexistence of EATDS and CF and exhibited the retrograde migration of the cells (EAT in green and CF in red) which was confirmed via z-stacks and 3D rendering (Figure 7d-i) (Supporting Information: Videos 1-3).…”

Epicardial Adipose Tissue and Diabetic Cardiomyopathy

“…As a member of the adipose-derived stem cell secretome, exosomes can drive the heterogeneous expression of stem cells, including inflammation and cell proliferation, which provides the possibility for cardiomyocyte regeneration (Figure 2). 79,80 Exosomes may be involved in the pathogenesis of DCM, but the specific mechanism remains unclear. It has been reported that exosomal microRNA can regulate pancreatic b-cell activity by regulating the intercellular signaling cascade, which is an essential determinant of pancreatic b-cell function integrity.…”