Abstract 14013: Direct Comparison of Disease-Specific versus TALEN-Corrected iPSC-Derived Cardiomyocytes from Dilated Cardiomyopathy Patients
Introduction: Recent advances in regenerative medicine for cardiovascular disease (CVD) therapy focus on delivery of autologous induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) to recover heart function without immunosuppression. Here, we perform a direct comparison of iPSC-CMs from a healthy individual, a DCM patient, and following genome-correction with transcription activator-like effector nucleases (TALENs) both in vitro and in vivo.
Methods & Results: We established patient-specific iPSC-CMs that recapitulate DCM disease-phenotypes in-vitro, including disrupted sarcomeres and impaired Ca2+ handling (p<0.05). We corrected the patient′s DCM-specific mutation R173W in the TNNT2 locus via TALEN-mediated footprint-free genome editing. Isogenic TALEN-corrected iPSC-CMs recovered functional properties comparable to healthy control. Ca2+ transient analysis revealed no significant difference between TALEN-corrected iPSC-CMs and healthy control regarding amplitude (ΔF/F0 4.71±0.55), decay (277.6 ±28.2 ms), and time to peak (240.5±22.5 ms). Confocal microscopy of TALEN-corrected iPSC-CMs confirmed sarcomeric structures as in healthy control (Fig 1). We transplanted (i) 1x10*7 DCM iPSC-CMs, (ii) 1x10*7 TALEN-corrected iPSC-CMs, and (iii) 1x10*7 healthy control iPSC-CMs into a subacute myocardial infarct (MI) rat model (n=15/group). Cell engraftment into the host myocardium was confirmed by immunohistochemistry. Comprehensive functional analysis via magnetic resonance imaging (MRI), echocardiography is underway.
Conclusions: We generated TALEN-corrected iPSC-CMs from a DCM patient, which recover in vitro functional parameters of healthy control iPSC-CMs. We transplanted for the first time patient-specific and TALEN-corrected iPSC-CMs into a rodent MI model. This approach may represent a viable option for mechanistic analysis and regenerative medicine in CVD patients. Figure 1:
Author Disclosures: A. Ebert: None. J. Riegler: None. I. Karakikes: None. V. Termglinchan: None. M. Mameen: None. H. Wu: None. Q. Shen: None. T. Yeh: None. J. Gold: None. J.C. Wu: None.
This research has received full or partial funding support from the American Heart Association.
- © 2014 by American Heart Association, Inc.