Regenerative Medicine of Cardiovascular Diseases

Human induced pluripotent stem cells (hiPSCs) are stem cells that are derived from the somatic cells (e.g. blood or skin cells) of healthy individuals or patients. They have the capacity to self-renew indefinitely and to give rise to any cell type of the body in vitro, making them an exceptionally powerful tool for studying human disease in patient-specific backgrounds. A major focus of our research is using hiPSC-based disease modelling to gain insights into the molecular and cellular mechanisms involved in cardiovascular disorders such as arrhythmogenic right ventricular cardiomyopathy (ARVC), hypoplastic left heart syndrome (HLHS), cardiomyopathy associated with Noonan syndrome, left ventricular non-compaction (LVNC), and others. Once a therapeutic target has been identified, we also use hiPSC-derived cells as a platform for testing novel therapies, as we have previously done for Duchenne muscular dystrophy (DMD).

Confocal immunofluorescence image of hiPSC-derived cardiomyocytes stained for the sarcomere proteins cardiac troponin T (red) and a-actinin (green). Scale bar: 50 µm.

Live immunofluorescence imaging of a 3D cardiac spheroid derived from a double reporter stem cell line expressing eGFP and mCherry in the endogenous loci of the cardiac transcription factors NKX2.5 and TBX5, respectively. Scale bar: 200 µm.

As cells cultured in 2D fail to recapitulate certain features of their in vivo counterparts, part of our work is dedicated to developing 3D cardiac models that better mimic the physiology of heart tissue. We routinely produce engineered heart tissue based on hiPSC-derived cardiac cells seeded on decellularized ventricular matrix from pigs or non-human primates, which we cultivate in a biomimetic chamber system promoting tissue maturation by continuous mechanical and electrical stimulation. We have also established an ex vivo embryo culture system that allows us to study the function of human cardiac cells in human-mouse or human-pig chimeras. Finally, we recently generated human pluripotent stem cell-derived cardiac organoids showing morphological, functional, and molecular self-organization of ventricular myocardium and epicardium.

Confocal immunofluorescence image of a cardiac organoid section showing the outer epicardial layer (tight junction protein 1 staining in yellow), the subjacent epicardium-derived cells (EPDCs) having undergone epithelial-to-mesenchymal transition (vimentin staining in magenta) and the myocardial layer (cardiac troponin T staining in green). Scale bar: 50 µm.

Biomimetic chamber containing engineered heart tissue undergoing radiofrequency ablation to mimic an acute myocardial injury.

• Generation, culture and differentiation of human pluripotent stem cells in 2D and 3D
• CRISPR/Cas9 gene editing for the generation of engineered reporter lines and knock-in/knock-out lines
• Functional characterization of cardiomyocytes via calcium imaging, optical action potential recordings, contractility measurements
• Bulk and single-cell RNA-sequencing and ATAC-sequencing
• Immunofluorescence analysis, confocal microscopy
• Flow cytometry and FACS sorting
• Cell injection in embryos and ex vivo embryo culture
• Molecular biology techniques including cloning, digital PCR
• Capillary Western Blot, chromatin immunoprecipitation

Scientific Projects:
We regularly offer projects for diploma, PhD and M.D. students.


Third-Party Funding:

• European Research Council
• German Research Foundation
• German Centre for Cardiovascular Research, Munich Heart

Alliance

• Else Kröner-Fresenius Foundation

Original work

Meier AB, Zawada D, De Angelis MT, Martens LD, Santamaria G, Zengerle S, Nowak-Imialek M, Kornherr J, Zhang F, Tian Q, Wolf CM, Kupatt C, Sahara M, Lipp P, Theis FJ, Gagneur J, Goedel A, Laugwitz KL, Dorn T, Moretti A. (2023) Epicardioid single-cell genomics uncovers principles of human epicardium biology in heart development and disease. Nat Biotechnol. doi: 10.1038/s41587-023-01718-7. PMID: 37012447

Zawada D, Kornherr J, Meier AB, Santamaria G, Dorn T, Nowak-Imialek M, Ortmann D, Zhang F, Lachmann M, Dreßen M, Ortiz M, Mascetti VL, Harmer SC, Nobles M, Tinker A, De Angelis MT, Pedersen RA, Grote P, Laugwitz KL, Moretti A, Goedel A. (2023) Retinoic acid signaling modulation guides in vitro specification of human heart field-specific progenitor pools. Nat Commun. 14(1):1722. doi: 10.1038/s41467-023-36764-x. PMID: 37012244

Poch CM, Foo KS, De Angelis MT, Jennbacken K, Santamaria G, Bähr A, Wang QD, Reiter F, Hornaschewitz N, Zawada D, Bozoglu T, My I, Meier A, Dorn T, Hege S, Lehtinen ML, Tsoi YL, Hovdal D, Hyllner J, Schwarz S, Sudhop S, Jurisch V, Sini M, Fellows MD, Cummings M, Clarke J, Baptista R, Eroglu E, Wolf E, Klymiuk N, Lu K, Tomasi R, Dendorfer A, Gaspari M, Parrotta E, Cuda G, Krane M, Sinnecker D, Hoppmann P, Kupatt C, Fritsche-Danielson R, Moretti A, Chien KR, Laugwitz KL. (2022) Migratory and anti-fibrotic programmes define the regenerative potential of human cardiac progenitors.

Nat Cell Biol. 24(5):659-671. doi: 10.1038/s41556-022-00899-8. PMID: 35550611

Esfandyari D, Idrissou BMG, Hennis K, Avramopoulos P, Dueck A, El-Battrawy I, Grüter L, Meier MA, Näger AC, Ramanujam D, Dorn T, Meitinger T, Hagl C, Milting H, Borggrefe M, Fenske S, Biel M, Dendorfer A, Sassi Y, Moretti A, Engelhardt S. (2022) MicroRNA-365 regulates human cardiac action potential duration. Nat Commun. 13(1):220. doi: 10.1038/s41467-021-27856-7. PMID: 35017523

Krane M, Dreßen M, Santamaria G, My I, Schneider CM, Dorn T, Laue S, Mastantuono E, Berutti R, Rawat H, Gilsbach R, Schneider P, Lahm H, Schwarz S, Doppler SA, Paige S, Puluca N, Doll S, Neb I, Brade T, Zhang Z, Abou-Ajram C, Northoff B, Holdt LM, Sudhop S, Sahara M, Goedel A, Dendorfer A, Tjong FVY, Rijlaarsdam ME, Cleuziou J, Lang N, Kupatt C, Bezzina C, Lange R, Bowles NE, Mann M, Gelb BD, Crotti L, Hein L, Meitinger T, Wu S, Sinnecker D, Gruber PJ, Laugwitz KL, Moretti A. (2021) Sequential Defects in Cardiac Lineage Commitment and Maturation Cause Hypoplastic Left Heart Syndrome. Circulation. 144(17):1409-1428. doi: 10.1161/CIRCULATIONAHA.121.056198. PMID: 34694888

Moretti A, Fonteyne L, Giesert F, Hoppmann P, Meier AB, Bozoglu T, Baehr A, Schneider CM, Sinnecker D, Klett K, Fröhlich T, Rahman FA, Haufe T, Sun S, Jurisch V, Kessler B, Hinkel R, Dirschinger R, Martens E, Jilek C, Graf A, Krebs S, Santamaria G, Kurome M, Zakhartchenko V, Campbell B, Voelse K, Wolf A, Ziegler T, Reichert S, Lee S, Flenkenthaler F, Dorn T, Jeremias I, Blum H, Dendorfer A, Schnieke A, Krause S, Walter MC, Klymiuk N, Laugwitz KL, Wolf E, Wurst W, Kupatt C (2020). Somatic gene editing ameliorates skeletal and cardiac muscle failure in pig and human models of Duchenne muscular dystrophy. Nat Med 26(2):207-214

Guo Y, Dorn T, Kühl SJ, Linnemann A, Rothe M, Pfister AS, Vainio S, Laugwitz KL, Moretti A*, Kühl M* (2019). The Wnt inhibitor Dkk1 is required for maintaining the normal cardiac differentiation program in Xenopus laevis. Dev Biol. 449:1-13.

Book chapters, Reviews and Editorials