Downloaded from https://academic.oup.com/eurheartj/article/43/Supplement_2/ehac544.

2881/6746332 by Istituto di Matematica Applicata e Tecnologie Informatiche "Enrico Magenes" del CNR user on 28 March 2023
Basic Science – Cardiac Biology and Physiology 2881

A large-scale functional high-throughput screening identifies miR-515 and miR-519e as potent

inducers of human iPSC-cardiomyocyte proliferation
H.V. Renikunta 1 , K. Lazarow 2 , Y. Gong 3 , P.C. Shukla 1 , H. Giral 1 , A. Kratzer 1 , V. Nageswaran 1 , L. Opitz 4 , F.B. Engel 5 , A. Haghikia 1 , F. Paneni 3 ,
J.P. Von Kries 2 , K. Streckfuss-Boemeke 6 , U. Landmesser 1 , P. Jakob 7
1
Charite - Campus Benjamin Franklin, Department of Cardiology, Berlin, Germany; 2 Max Delbruck Center for Molecular Medicine, Leibniz-Institute
for Molecular Pharmacology, Berlin, Germany; 3 University of Zurich, Center for Molecular Cardiology, Schlieren, Switzerland; 4 University of Zurich,
Functional Genomics Center Zurich UZH/ETH, Zurich, Switzerland; 5 Friedrich Alexander University, Experimental Renal and Cardiovascular
Research, Department of Nephropathology, Erlangen, Germany; 6 University Medical Center of Gottingen (UMG), Clinic for Cardiology and
Pneumology, Goettingen, Germany; 7 University Heart Center, Cardiology, University Hospital Zurich, Zurich, Switzerland
Funding Acknowledgement: Type of funding sources: Foundation. Main funding source(s): This work was supported by the German Centre for
Cardiovascular Research (DZHK), Deutsche Stiftung für Herzforschung (DSHF) and OPO Foundation.

Introduction: Ischemic heart failure persists as a global health problem
despite optimized medical and adjunctive device therapies. Loss of car-
diomyocytes in the absence of a proliferative response comprise a major
contributor to pathological remodeling and death in this patient population.
Experimental studies have shown that microRNAs (miRNAs) may be used
as a therapeutic option to reinduce adult cardiomyocyte proliferation.
Purpose: This study thought to evaluate proliferative potential in human
cardiomyocytes after overexpression and inhibition of 2019 miRNAs.
Methods: To identify miRNAs that regulate cardiomyocyte proliferation, we
performed functional high-throughput screenings in human iPSC-derived
cardiomyocytes (hiPSC-CM) after transient hypoxia. Herein, 2019 miRNA-
mimics for overexpression and 2019 anti-miRs for inhibition were individ-
ually transfected to examine EdU-incorporation in hiPSC-CM. MiR-mimic-
515 and miR-mimic-519e that induced the highest EdU-uptake, were fur-
ther assessed by immunostaining and molecular methods for markers in-
dicative of early and late mitosis. In addition, RNA-Sequencing in hiPSC-
CM after overexpression of miR-515 and miR-519e was performed to ex-
amine differential gene expression and miRNA-modulated pathways in-
volved in cardiomyocyte proliferation.
Results: Using a functional high-throughput screening, we assessed dif-
ferential proliferative potential of 2019 miRNAs after transient hypoxia by
transfecting both miR-inhibitor and miR-mimic libraries in human iPSC-
derived cardiomyocytes (hiPSC-CM). Overexpression of 28 miRNAs sub-
stantially induced proliferative activity in hiPSC-CM, with an overrepresen-
tation of miRNAs belonging to the C19MC-cluster and adjacent miR-371–
373 family. Two of these miRNAs, miR-515 and miR-519e increased mark-
ers of early and late mitosis, with an additive cardiomyocyte turnover after
transient hypoxia and substantially increased Aurora B-kinase activity in
midbodies, indicative of cell division. These findings were supported by
molecular studies using qRT-PCR, Western blot, and RNA-Sequencing af-
ter overexpression of miR-515 and miR-519e showing substantial alter-
ations of signaling pathways relevant for cardiomyocytes proliferation in
human iPSC-CM.
Conclusion: Collectively, these results support a critical role of miR-515
and miR-519e for induction of proliferation in human cardiomyocytes un-
der hypoxic conditions, such as present in patients with ischemia-driven
cardiomyopathy.

ESC Congress 2022 – Barcelona, Spain

26 – 29 August 2022