Single-cell transcriptomics reveals an angiogenic cell population for therapeutic angiogenesis in adipose tissue

Abstract Background Therapeutic angiogenesis mediated by stem/progenitor cells is an attractive therapeutic option against cardiovascular disease (CVD). Adipose tissue (AT) can be safely obtained even in CVD patients with anti-platelet medications, and it a readily available source of culture-expanded adipose-derived stem (ADSCs) for transplantation. Single-cell transcriptome enables us to screen all the surface markers at once, while conventional strategies have been limited number target markers. Furthermore, gene profiling single-cell resolution used quantification each marker how many favorable purified without mixing detrimental cells. Purpose We aimed identify characterize cell population vivo angiogenic potential RNA sequencing (scRNA-seq) analysis xenograft experiments. Methods revisited scRNA-seq datasets single fraction from AT, bone-marrow (BM), umbilical-cord blood (UCB, n=6/organ) find populations pro-angiogenic potential. Next, we collected AT (n=23) multicolor flow cytometry quantify sort specific populations. PBS, marker-negative unsorted ADSCs were as controls. Xenograft models PKH26 pre-labeled human ADSC transplantation limb ischemia evaluate lectin capillary density, PKH+ engrafted ADSCs, recovery. Results Clustering divided CD45–CD31–CD34+ progenitor into 3 clusters. identified pro-/anti-angiogenic clusters based on expressions well-known factors. All genes encoding cell-surface proteins compared this functional clustering, resulted 17 screened (Fig. 1A, B). Taken together enrichment analysis, CD271+ showed predominant profile other top candidates including CD36 CD54 1C, D). evaluated sources BM UCB. Surprisingly, CD271 expressing significantly lower did not show UCB 2A). In 23 patients, decreased donor insulin resistance 2B). Cell therapy using demonstrated capacity those CD271– PBS model. enhanced efficacy same donors 2C–E). Conclusion study, through would most suitable isolation. promising could contribute better cell-based tackling CVD. Funding Acknowledgement Type funding sources: Public grant(s) – National budget only. Main source(s): Japan Society Promotion Science (JSPS) KAKENHI (Tokyo, Japan)