TY - JOUR
T1 - Bioactivity and miRNome Profiling of Native Extracellular Vesicles in Human Induced Pluripotent Stem Cell-Cardiomyocyte Differentiation
AU - Louro, Ana F.
AU - Paiva, Marta A.
AU - Oliveira, Marta R.
AU - Kasper, Katharina A.
AU - Alves, Paula M.
AU - Gomes-Alves, Patrícia
AU - Serra, Margarida
N1 - Funding Information:
The authors acknowledge A.L. Sousa and E.M. Tranfield from the Electron Microscopy Facility at the IGC for the transmission electron microscopy work. The authors also thank IGC's histopathology unit for the aggregate cryosections. The authors thank Prof Joost Sluijter for the helpful discussions regarding this work. This work was supported by EU‐funded project BRAV3 (H2020, ID:874827); EU Interreg Sudoe – funded project CardioPatch (SOE4/P1/E1063); Fundação para a Ciência e Tecnologia (FCT)‐funded projects NETDIAMOND (SAICTPAC/0047/2015) and MetaCardio (PTDC/BTMSAL/32566/2017). iNOVA4Health – UIDB/04462/2020 and UIDP/04462/2020, a program financially supported by FCT / Ministério da Ciência, Tecnologia e Ensino Superior, through national funds, is also acknowledged. A.F.L. was financed by FCT under Grant No. PD/BD/139078/2018.
Funding Information:
The authors acknowledge A.L. Sousa and E.M. Tranfield from the Electron Microscopy Facility at the IGC for the transmission electron microscopy work. The authors also thank IGC's histopathology unit for the aggregate cryosections. The authors thank Prof Joost Sluijter for the helpful discussions regarding this work. This work was supported by EU-funded project BRAV3 (H2020, ID:874827); EU Interreg Sudoe – funded project CardioPatch (SOE4/P1/E1063); Fundação para a Ciência e Tecnologia (FCT)-funded projects NETDIAMOND (SAICTPAC/0047/2015) and MetaCardio (PTDC/BTMSAL/32566/2017). iNOVA4Health – UIDB/04462/2020 and UIDP/04462/2020, a program financially supported by FCT / Ministério da Ciência, Tecnologia e Ensino Superior, through national funds, is also acknowledged. A.F.L. was financed by FCT under Grant No. PD/BD/139078/2018.
Publisher Copyright:
© 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.
PY - 2022/5/25
Y1 - 2022/5/25
N2 - Extracellular vesicles (EV) are an attractive therapy to boost cardiac regeneration. Nevertheless, identification of native EV and corresponding cell platform(s) suitable for therapeutic application, is still a challenge. Here, EV are isolated from key stages of the human induced pluripotent stem cell-cardiomyocyte (hiPSC-CM) differentiation and maturation, i.e., from hiPSC (hiPSC-EV), cardiac progenitors, immature and mature cardiomyocytes, with the aim of identifying a promising cell biofactory for EV production, and pinpoint the genetic signatures of bioactive EV. EV secreted by hiPSC and cardiac derivatives show a typical size distribution profile and the expression of specific EV markers. Bioactivity assays show increased tube formation and migration in HUVEC treated with hiPSC-EV compared to EV from committed cell populations. hiPSC-EV also significantly increase cell cycle activity of hiPSC-CM. Global miRNA expression profiles, obtained by small RNA-seq analysis, corroborate an EV-miRNA pattern indicative of stem cell to cardiomyocyte specification, confirming that hiPSC-EV are enriched in pluripotency-associated miRNA with higher in vitro pro-angiogenic and pro-proliferative properties. In particular, a stemness maintenance miRNA cluster upregulated in hiPSC-EV targets the PTEN/PI3K/AKT pathway, involved in cell proliferation and survival. Overall, the findings validate hiPSC as cell biofactories for EV production for cardiac regenerative applications.
AB - Extracellular vesicles (EV) are an attractive therapy to boost cardiac regeneration. Nevertheless, identification of native EV and corresponding cell platform(s) suitable for therapeutic application, is still a challenge. Here, EV are isolated from key stages of the human induced pluripotent stem cell-cardiomyocyte (hiPSC-CM) differentiation and maturation, i.e., from hiPSC (hiPSC-EV), cardiac progenitors, immature and mature cardiomyocytes, with the aim of identifying a promising cell biofactory for EV production, and pinpoint the genetic signatures of bioactive EV. EV secreted by hiPSC and cardiac derivatives show a typical size distribution profile and the expression of specific EV markers. Bioactivity assays show increased tube formation and migration in HUVEC treated with hiPSC-EV compared to EV from committed cell populations. hiPSC-EV also significantly increase cell cycle activity of hiPSC-CM. Global miRNA expression profiles, obtained by small RNA-seq analysis, corroborate an EV-miRNA pattern indicative of stem cell to cardiomyocyte specification, confirming that hiPSC-EV are enriched in pluripotency-associated miRNA with higher in vitro pro-angiogenic and pro-proliferative properties. In particular, a stemness maintenance miRNA cluster upregulated in hiPSC-EV targets the PTEN/PI3K/AKT pathway, involved in cell proliferation and survival. Overall, the findings validate hiPSC as cell biofactories for EV production for cardiac regenerative applications.
KW - cardiac regeneration
KW - cell biofactory
KW - extracellular vesicles
KW - miRNome
KW - small RNA-seq
UR - http://www.scopus.com/inward/record.url?scp=85126943483&partnerID=8YFLogxK
U2 - 10.1002/advs.202104296
DO - 10.1002/advs.202104296
M3 - Article
C2 - 35322574
AN - SCOPUS:85126943483
SN - 2198-3844
VL - 9
JO - Advanced Science
JF - Advanced Science
IS - 15
M1 - 2104296
ER -