Abstract
Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) are powerful in vitro models to study the mechanisms underlying cardiomyopathies and cardiotoxicity. Quantification of the contractile function in single hiPSC-CMs at high-throughput and over time is essential to disentangle how cellular mechanisms affect heart function. Here, we present CONTRAX, an open-access, versatile, and streamlined pipeline for quantitative tracking of the contractile dynamics of single hiPSC-CMs over time. Three software modules enable: parameter-based identification of single hiPSC-CMs; automated video acquisition of >200 cells/hour; and contractility measurements via traction force microscopy. We analyze >4,500 hiPSC-CMs over time in the same cells under orthogonal conditions of culture media and substrate stiffnesses; +/− drug treatment; +/− cardiac mutations. Using undirected clustering, we reveal converging maturation patterns, quantifiable drug response to Mavacamten and significant deficiencies in hiPSC-CMs with disease mutations. CONTRAX empowers researchers with a potent quantitative approach to develop cardiac therapies.
| Original language | English |
|---|---|
| Article number | 5427 |
| Journal | Nature Communications |
| Volume | 15 |
| Issue number | 1 |
| DOIs | |
| State | Published - Dec 2024 |
Bibliographical note
Publisher Copyright:© The Author(s) 2024.
Funding
We thank all members of the Pruitt laboratory at Stanford University and UC Santa Barbara and members of the Blau laboratory at Stanford University for helpful discussions and support. Wild-type hiPSCs (WTC cell line) were obtained as a generous gift from collaborators, and from the Allen Cell Collection36,38,78. This research was supported by the Swiss National Science Foundation (SNSF) Early Postdoc Mobility Fellowship (#P2SKP2_164954 to G.P.) and Postdoc Mobility Fellowship (#P400PM_180825 to G.P.), the American Heart Association (AHA Award 18POST34080160 to G.P., 20POST35211011 to A.S.V.R., and 17CSA33590101 to H.M.B. and B.L.P.); the National Institutes of Health (NIH 1R21HL13099301 and RM1GM131981 to B.L.P., and K99HL153679 to A.S.V.R); and the Baxter Foundation, Li Ka Shing Foundation and The Stanford Cardiovascular Institute to H.M.B. We also acknowledge support from the National Science Foundation GRFP (to A.K.D. and R.E.W., E. A. C.), the Stanford Office of the Vice Provost for Graduate Education (to A.K.D.), Ford Foundation Pre-doctoral Fellowship (E. A. C.); the Stanford Bio-X Summer Undergraduate Research Program (to F. Birnbaum), a Major Grant from the Stanford University Vice Provost for Undergraduate Education (to F.B.), and NIH 1F31HL158227 (O.C.). This research was also supported by the National Natural Science Foundation of China (82070248 to A.C.Y.C.) and Shanghai Pujiang Program (19PJ1407000 to A.C.Y.C.); the Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning (0900000024 to A.C.Y.C.); Innovative Research Team of High-Level Local Universities in Shanghai (A.C.Y.C.); the American Heart Association (13POST14480004 and 18CDA34110411 to A.C.Y.C.); and the Canadian Institutes of Health Research Fellowship (201411MFE-338745-169197 to A.C.Y.C.). This research was supported by the Swiss National Science Foundation (SNSF) Early Postdoc Mobility Fellowship (#P2SKP2_164954 to G.P.) and Postdoc Mobility Fellowship (#P400PM_180825 to G.P.), the American Heart Association (AHA Award 18POST34080160 to G.P., 20POST35211011 to A.S.V.R., and 17CSA33590101 to H.M.B. and B.L.P.); the National Institutes of Health (NIH 1R21HL13099301 and RM1GM131981 to B.L.P., and K99HL153679 to A.S.V.R); and the Baxter Foundation, Li Ka Shing Foundation and The Stanford Cardiovascular Institute to H.M.B. We also acknowledge support from the National Science Foundation GRFP (to A.K.D. and R.E.W., E. A. C.), the Stanford Office of the Vice Provost for Graduate Education (to A.K.D.), Ford Foundation Pre-doctoral Fellowship (E. A. C.); the Stanford Bio-X Summer Undergraduate Research Program (to F. Birnbaum), a Major Grant from the Stanford University Vice Provost for Undergraduate Education (to F.B.), and NIH 1F31HL158227 (O.C.). This research was also supported by the National Natural Science Foundation of China (82070248 to A.C.Y.C.) and Shanghai Pujiang Program (19PJ1407000 to A.C.Y.C.); the Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning (0900000024 to A.C.Y.C.); Innovative Research Team of High-Level Local Universities in Shanghai (A.C.Y.C.); the American Heart Association (13POST14480004 and 18CDA34110411 to A.C.Y.C.); and the Canadian Institutes of Health Research Fellowship (201411MFE-338745-169197 to A.C.Y.C.).
| Funders | Funder number |
|---|---|
| Donald E. and Delia B. Baxter Foundation | |
| National Science Foundation Arctic Social Science Program | |
| Ford Foundation | |
| Stanford Cardiovascular Institute, School of Medicine, Stanford University | |
| Li Ka Shing Foundation | |
| Office of the Vice Provost for Graduate Education, Stanford University | |
| University of California, Santa Barbara | |
| National Natural Science Foundation of China (NSFC) | 82070248 |
| Not added | 164954 |
| Shanghai Pujiang Program, China | 19PJ1407000 |
| American the American Heart Association | 20POST35211011, 18POST34080160, 17CSA33590101 |
| Innovative Research Team of High-level Local University in Shanghai | 13POST14480004, 18CDA34110411 |
| National Institutes of Health (NIH) | 1R21HL13099301, RM1GM131981, K99HL153679 |
| Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning | 0900000024 |
| Stanford University Vice Provost for Undergraduate Education | 1F31HL158227 |
| Canadian Institutes of Health Research | 201411MFE-338745-169197 |
ASJC Scopus subject areas
- General Chemistry
- General Biochemistry, Genetics and Molecular Biology
- General Physics and Astronomy
