Notch pathway activation enhances cardiosphere in vitro expansion

TY - JOUR

T1 - Notch pathway activation enhances cardiosphere in vitro expansion

AU - Secco, Ilaria

AU - Barile, Lucio

AU - Torrini, Consuelo

AU - Zentilin, Lorena

AU - Vassalli, Giuseppe

AU - Giacca, Mauro

AU - Collesi, Chiara

PY - 2018/11/1

Y1 - 2018/11/1

N2 - Cardiospheres (CSps) are self-assembling clusters of a heterogeneous population of poorly differentiated cells outgrowing from in vitro cultured cardiac explants. Scanty information is available on the molecular pathways regulating CSp growth and their differentiation potential towards cardiac and vascular lineages. Here we report that Notch1 stimulates a massive increase in both CSp number and size, inducing a peculiar gene expression programme leading to a cardiovascular molecular signature. These effects were further enhanced using Adeno-Associated Virus (AAV)-based gene transfer of activated Notch1-intracellular domain (N1-ICD) or soluble-Jagged1 (sJ1) ligand to CSp-forming cells. A peculiar effect was exploited by selected pro-proliferating miRNAs: hsa-miR-590-3p induced a cardiovascular gene expression programme, while hsa-miR-199a-3p acted as the most potent stimulus for the activation of the Notch pathway, thus showing that, unlike in adult cardiomyocytes, these miRNAs involve Notch signalling activation in CSps. Our results identify Notch1 as a crucial regulator of CSp growth and differentiation along the vascular lineage, raising the attracting possibility that forced activation of this pathway might be exploited to promote in vitro CSp expansion as a tool for toxicology screening and cell-free therapeutic strategies.

AB - Cardiospheres (CSps) are self-assembling clusters of a heterogeneous population of poorly differentiated cells outgrowing from in vitro cultured cardiac explants. Scanty information is available on the molecular pathways regulating CSp growth and their differentiation potential towards cardiac and vascular lineages. Here we report that Notch1 stimulates a massive increase in both CSp number and size, inducing a peculiar gene expression programme leading to a cardiovascular molecular signature. These effects were further enhanced using Adeno-Associated Virus (AAV)-based gene transfer of activated Notch1-intracellular domain (N1-ICD) or soluble-Jagged1 (sJ1) ligand to CSp-forming cells. A peculiar effect was exploited by selected pro-proliferating miRNAs: hsa-miR-590-3p induced a cardiovascular gene expression programme, while hsa-miR-199a-3p acted as the most potent stimulus for the activation of the Notch pathway, thus showing that, unlike in adult cardiomyocytes, these miRNAs involve Notch signalling activation in CSps. Our results identify Notch1 as a crucial regulator of CSp growth and differentiation along the vascular lineage, raising the attracting possibility that forced activation of this pathway might be exploited to promote in vitro CSp expansion as a tool for toxicology screening and cell-free therapeutic strategies.

KW - Adeno-associated virus (AAV)

KW - Cardiac Progenitors

KW - Micro-RNAs

KW - Notch

UR - http://www.scopus.com/inward/record.url?scp=85052377045&partnerID=8YFLogxK

U2 - 10.1111/jcmm.13832

DO - 10.1111/jcmm.13832

M3 - Article

C2 - 30138533

AN - SCOPUS:85052377045

SN - 1582-1838

VL - 22

SP - 5583

EP - 5595

JO - JOURNAL OF CELLULAR AND MOLECULAR MEDICINE

JF - JOURNAL OF CELLULAR AND MOLECULAR MEDICINE

IS - 11

ER -