Gene editing to prevent ventricular arrhythmias associated with cardiomyocyte cell therapy

  • Cell Stem Cell. 2023 Apr 6;30(4):396-414.e9. doi: 10.1016/j.stem.2023.03.010.
Silvia Marchiano  1 ,  Kenta Nakamura  2 ,  Hans Reinecke  1 ,  Lauren Neidig  3 ,  Michael Lai  4 ,  Shin Kadota  5 ,  Filippo Perbellini  4 ,  Xiulan Yang  1 ,  Jordan M Klaiman  1 ,  Leslie P Blakely  1 ,  Elaheh Karbassi  1 ,  Paul A Fields  6 ,  Aidan M Fenix  1 ,  Kevin M Beussman  7 ,  Anu Jayabalu  8 ,  Faith A Kalucki  8 ,  Jennifer C Potter  8 ,  Akiko Futakuchi-Tsuchida  8 ,  Gerhard J Weber  2 ,  Sarah Dupras  8 ,  Hiroshi Tsuchida  8 ,  Lil Pabon  8 ,  Lili Wang  9 ,  Björn C Knollmann  9 ,  Steven Kattman  8 ,  R Scott Thies  8 ,  Nathan Sniadecki  10 ,  W Robb MacLellan  2 ,  Alessandro Bertero  1 ,  Charles E Murry  11
Affiliations
  • 1. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Department of Laboratory Medicine & Pathology, University of Washington, Seattle, WA 98195, USA.
  • 2. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA 98195, USA.
  • 3. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Department of Comparative Medicine, University of Washington, Seattle, WA 98195, USA.
  • 4. Sana Biotechnology, Seattle, WA 98102, USA.
  • 5. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Department of Laboratory Medicine & Pathology, University of Washington, Seattle, WA 98195, USA; Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA 98195, USA; Department of Regenerative Science and Medicine, Shinshu University, Matsumoto 390-8621, Japan.
  • 6. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Department of Laboratory Medicine & Pathology, University of Washington, Seattle, WA 98195, USA; Adaptive Biotechnologies, Seattle, WA 98102, USA.
  • 7. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Department of Mechanical Engineering, University of Washington, 3720 15(th) Avenue NE, Seattle, WA 98105, USA.
  • 8. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Department of Laboratory Medicine & Pathology, University of Washington, Seattle, WA 98195, USA; Sana Biotechnology, Seattle, WA 98102, USA.
  • 9. Division of Clinical Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
  • 10. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Department of Laboratory Medicine & Pathology, University of Washington, Seattle, WA 98195, USA; Department of Mechanical Engineering, University of Washington, 3720 15(th) Avenue NE, Seattle, WA 98105, USA; Department of Bioengineering, University of Washington, Seattle, WA 98195, USA.
  • 11. Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Brotman Building Room 453, Seattle, WA 98109, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA 98109, USA; Department of Laboratory Medicine & Pathology, University of Washington, Seattle, WA 98195, USA; Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA 98195, USA; Sana Biotechnology, Seattle, WA 98102, USA; Department of Bioengineering, University of Washington, Seattle, WA 98195, USA. Electronic address: [email protected].
PMID: 37028405 DOI: 10.1016/j.stem.2023.03.010
Abstract

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) offer a promising cell-based therapy for Myocardial Infarction. However, the presence of transitory ventricular Arrhythmias, termed engraftment Arrhythmias (EAs), hampers clinical applications. We hypothesized that EA results from pacemaker-like activity of hPSC-CMs associated with their developmental immaturity. We characterized ion channel expression patterns during maturation of transplanted hPSC-CMs and used pharmacology and genome editing to identify those responsible for automaticity in vitro. Multiple engineered cell lines were then transplanted in vivo into uninjured porcine Hearts. Abolishing depolarization-associated genes HCN4, CACNA1H, and SLC8A1, along with overexpressing hyperpolarization-associated KCNJ2, creates hPSC-CMs that lack automaticity but contract when externally stimulated. When transplanted in vivo, these cells engrafted and coupled electromechanically with host cardiomyocytes without causing sustained EAs. This study supports the hypothesis that the immature electrophysiological prolife of hPSC-CMs mechanistically underlies EA. Thus, targeting automaticity should improve the safety profile of hPSC-CMs for cardiac remuscularization.

Keywords
arrhythmia; automaticity; cardiac remuscularization; cell therapy; engraftment arrhythmia; hPSC-CM maturation; heart regeneration; human pluripotent stem cell-derived cardiomyocytes; myocardial infarction; pacemaker.