Advancing physiological maturation in human induced pluripotent stem cell-derived cardiac muscle by gene editing an inducible adult troponin isoform switch
| Autor: | Fikru B. Bedada, Joseph M. Metzger, Jennifer L. Mikkila, Matthew Wheelwright, Mohammad A. Mandegar, Brian R. Thompson |
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| Rok vydání: | 2020 |
| Předmět: |
Adult
0301 basic medicine Gene isoform Induced Pluripotent Stem Cells Biology Sarcomere Article Cell Line 03 medical and health sciences 0302 clinical medicine Myosin medicine Humans Protein Isoforms Myocyte Myocytes Cardiac Induced pluripotent stem cell Gene Editing Genome Human Myocardium Troponin I Cardiac muscle Reproducibility of Results Cell Differentiation Cell Biology Myocardial Contraction Troponin Cell biology Phospholamban 030104 developmental biology medicine.anatomical_structure Gene Expression Regulation cardiovascular system biology.protein Molecular Medicine 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Stem Cells |
| ISSN: | 1549-4918 1066-5099 |
| DOI: | 10.1002/stem.3235 |
| Popis: | Advancing maturation of stem cell-derived cardiac muscle represents a major barrier to progress in cardiac regenerative medicine. Cardiac muscle maturation involves a myriad of gene, protein, and cell-based transitions, spanning across all aspects of cardiac muscle form and function. We focused here on a key developmentally controlled transition in the cardiac sarcomere, the functional unit of the heart. Using a gene-editing platform, human induced pluripotent stem cell (hiPSCs) were engineered with a drug-inducible expression cassette driving the adult cardiac troponin I (cTnI) regulatory isoform, a transition shown to be a rate-limiting step in advancing sarcomeric maturation of hiPSC cardiac muscle (hiPSC-CM) toward the adult state. Findings show that induction of the adult cTnI isoform resulted in the physiological acquisition of adult-like cardiac contractile function in hiPSC-CMs in vitro. Specifically, cTnI induction accelerated relaxation kinetics at baseline conditions, a result independent of alterations in the kinetics of the intracellular Ca2+ transient. In comparison, isogenic unedited hiPSC-CMs had no cTnI induction and no change in relaxation function. Temporal control of adult cTnI isoform induction did not alter other developmentally regulated sarcomere transitions, including myosin heavy chain isoform expression, nor did it affect expression of SERCA2a or phospholamban. Taken together, precision genetic targeting of sarcomere maturation via inducible TnI isoform switching enables physiologically relevant adult myocardium-like contractile adaptations that are essential for beat-to-beat modulation of adult human heart performance. These findings have relevance to hiPSC-CM structure-function and drug-discovery studies in vitro, as well as for potential future clinical applications of physiologically optimized hiPSC-CM in cardiac regeneration/repair. |
| Databáze: | OpenAIRE |
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