PMID- 38534508 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20240330 IS - 2306-5354 (Print) IS - 2306-5354 (Electronic) IS - 2306-5354 (Linking) VI - 11 IP - 3 DP - 2024 Feb 28 TI - Contractile and Genetic Characterization of Cardiac Constructs Engineered from Human Induced Pluripotent Stem Cells: Modeling of Tuberous Sclerosis Complex and the Effects of Rapamycin. LID - 10.3390/bioengineering11030234 [doi] LID - 234 AB - The implementation of three-dimensional tissue engineering concurrently with stem cell technology holds great promise for in vitro research in pharmacology and toxicology and modeling cardiac diseases, particularly for rare genetic and pediatric diseases for which animal models, immortal cell lines, and biopsy samples are unavailable. It also allows for a rapid assessment of phenotype-genotype relationships and tissue response to pharmacological manipulation. Mutations in the TSC1 and TSC2 genes lead to dysfunctional mTOR signaling and cause tuberous sclerosis complex (TSC), a genetic disorder that affects multiple organ systems, principally the brain, heart, skin, and kidneys. Here we differentiated healthy (CC3) and tuberous sclerosis (TSP8-15) human induced pluripotent stem cells (hiPSCs) into cardiomyocytes to create engineered cardiac tissue constructs (ECTCs). We investigated and compared their mechano-elastic properties and gene expression and assessed the effects of rapamycin, a potent inhibitor of the mechanistic target of rapamycin (mTOR). The TSP8-15 ECTCs had increased chronotropy compared to healthy ECTCs. Rapamycin induced positive inotropic and chronotropic effects (i.e., increased contractility and beating frequency, respectively) in the CC3 ECTCs but did not cause significant changes in the TSP8-15 ECTCs. A differential gene expression analysis revealed 926 up- and 439 down-regulated genes in the TSP8-15 ECTCs compared to their healthy counterparts. The application of rapamycin initiated the differential expression of 101 and 31 genes in the CC3 and TSP8-15 ECTCs, respectively. A gene ontology analysis showed that in the CC3 ECTCs, the positive inotropic and chronotropic effects of rapamycin correlated with positively regulated biological processes, which were primarily related to the metabolism of lipids and fatty and amino acids, and with negatively regulated processes, which were predominantly associated with cell proliferation and muscle and tissue development. In conclusion, this study describes for the first time an in vitro TSC cardiac tissue model, illustrates the response of normal and TSC ECTCs to rapamycin, and provides new insights into the mechanisms of TSC. FAU - Sidorov, Veniamin Y AU - Sidorov VY AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN 37235, USA. AD - Department of Biomedical Engineering, Vanderbilt University, Nashville, TN 37235, USA. FAU - Sidorova, Tatiana N AU - Sidorova TN AD - Department of Anesthesiology, Vanderbilt University Medical Center, Nashville, TN 37232, USA. FAU - Samson, Philip C AU - Samson PC AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN 37235, USA. AD - Department of Physics and Astronomy, Vanderbilt University, Nashville, TN 37212, USA. FAU - Reiserer, Ronald S AU - Reiserer RS AUID- ORCID: 0000-0002-3786-7893 AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN 37235, USA. AD - Department of Physics and Astronomy, Vanderbilt University, Nashville, TN 37212, USA. FAU - Britt, Clayton M AU - Britt CM AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN 37235, USA. AD - Department of Physics and Astronomy, Vanderbilt University, Nashville, TN 37212, USA. FAU - Neely, M Diana AU - Neely MD AD - Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA. FAU - Ess, Kevin C AU - Ess KC AD - Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA. FAU - Wikswo, John P AU - Wikswo JP AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN 37235, USA. AD - Department of Biomedical Engineering, Vanderbilt University, Nashville, TN 37235, USA. AD - Department of Physics and Astronomy, Vanderbilt University, Nashville, TN 37212, USA. AD - Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA. LA - eng GR - A68485/NH/NIH HHS/United States GR - RR030956/NH/NIH HHS/United States GR - P30 DK020593/DK/NIDDK NIH HHS/United States GR - DK58404/NH/NIH HHS/United States GR - P30 CA068485/CA/NCI NIH HHS/United States GR - DK20593/NH/NIH HHS/United States GR - U24 DK059637/DK/NIDDK NIH HHS/United States GR - 1S10OD021630 1/NH/NIH HHS/United States GR - P30 DK058404/DK/NIDDK NIH HHS/United States GR - S10 OD021630/OD/NIH HHS/United States GR - RR024975/NH/NIH HHS/United States GR - DK59637/NH/NIH HHS/United States GR - G20 RR030956/RR/NCRR NIH HHS/United States GR - P30 EY008126/EY/NEI NIH HHS/United States GR - UH3TR002097,/NH/NIH HHS/United States GR - UH3 TR002097/TR/NCATS NIH HHS/United States GR - UL1 RR024975/RR/NCRR NIH HHS/United States PT - Journal Article DEP - 20240228 PL - Switzerland TA - Bioengineering (Basel) JT - Bioengineering (Basel, Switzerland) JID - 101676056 PMC - PMC10968530 OTO - NOTNLM OT - I-Wire cardiac construct OT - heart-on-a-chip OT - hiPSC OT - mTOR OT - rapamycin OT - tuberous sclerosis complex COIS- The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EDAT- 2024/03/27 12:45 MHDA- 2024/03/27 12:46 PMCR- 2024/02/28 CRDT- 2024/03/27 09:25 PHST- 2024/01/12 00:00 [received] PHST- 2024/02/21 00:00 [revised] PHST- 2024/02/23 00:00 [accepted] PHST- 2024/03/27 12:46 [medline] PHST- 2024/03/27 12:45 [pubmed] PHST- 2024/03/27 09:25 [entrez] PHST- 2024/02/28 00:00 [pmc-release] AID - bioengineering11030234 [pii] AID - bioengineering-11-00234 [pii] AID - 10.3390/bioengineering11030234 [doi] PST - epublish SO - Bioengineering (Basel). 2024 Feb 28;11(3):234. doi: 10.3390/bioengineering11030234.