Electrophysiological engineering of heart-derived cells with calcium-dependent potassium channels improves cell therapy efficacy for cardioprotection

Patrick Vigneault,Stanley Nattel,Connor Michie,Pushpinder Kanda,Darryl R Davis,Sandrine Parent

doi:10.1038/s41467-021-25180-8

Abstract

We have shown that calcium-activated potassium (KCa)-channels regulate fundamental progenitor-cell functions, including proliferation, but their contribution to cell-therapy effectiveness is unknown. Here, we test the participation of KCa-channels in human heart explant-derived cell (EDC) physiology and therapeutic potential. TRAM34-sensitive KCa3.1-channels, encoded by the KCNN4 gene, are exclusively expressed in therapeutically bioactive EDC subfractions and maintain a strongly polarized resting potential; whereas therapeutically inert EDCs lack KCa3.1 channels and exhibit depolarized resting potentials. Somatic gene transfer of KCNN4 results in membrane hyperpolarization and increases intracellular [Ca2+], which boosts cell-proliferation and the production of pro-healing cytokines/nanoparticles. Intramyocardial injection of EDCs after KCNN4-gene overexpression markedly increases the salutary effects of EDCs on cardiac function, viable myocardium and peri-infarct neovascularization in a well-established murine model of ischemic cardiomyopathy. Thus, electrophysiological engineering provides a potentially valuable strategy to improve the therapeutic value of progenitor cells for cardioprotection and possibly other indications.

Highlights

We have shown that calcium-activated potassium (KCa)-channels regulate fundamental progenitor-cell functions, including proliferation, but their contribution to cell-therapy effectiveness is unknown
Stem cell therapy has emerged as a potential approach to prevent the progression of heart failure[2] and, among the number of candidates proposed for cardiac cell therapy, heart explant-derived cells (EDCs) have been developed as a promising paracrine-based cell therapeutic[3,4]
We found that the therapeutic potential of EDCs derived from human hearts is governed by the expression and function of the intermediate-conductance Ca2+-activated channel KCa3.1

Summary

Introduction

We have shown that calcium-activated potassium (KCa)-channels regulate fundamental progenitor-cell functions, including proliferation, but their contribution to cell-therapy effectiveness is unknown. We recently showed that the function of bone-marrow-derived mesenchymal stem cells and resident cardiac c-Kit+ cells is critically governed by the intermediate-conductance Ca2+-activated K+ channel KCa3.1 (encoded by the KCNN4 gene)[10] In both adult progenitor cell types, KCa3.1 channels open in response to store-operated Ca2+-entry (SOCE) to hyperpolarize the cell membrane, increase the driving force for Ca2+ entry, and enhance transmembrane Ca2+ flux. We hypothesized that KCa3.1-channel activity might influence the behavior of therapeutically relevant cells, and that KCNN4 overexpression might improve therapeutic efficacy by optimizing Vmem during SOCE We test this hypothesis and provide evidence that tailoring plasma-membrane ion-channel function influences the therapeutic efficacy of ex vivo expanded human heart cells using a promising adult cell therapeutic within an established immunodeficient mouse model of ischemic cardiomyopathy

Methods

Results

Conclusion