Pathologic Stimulus Determines Lineage Commitment of Cardiac C-kit+ Cells

Zhongming Chen; Wuqiang Zhu; Ingrid Bender; Wuming Gong; Il-Youp Kwak; Amritha Yellamilli; Thomas J Hodges; Natsumi Nemoto; Jianyi Zhang; Daniel J Garry; Jop H van Berlo

doi:10.1161/CIRCULATIONAHA.117.030137

Pathologic Stimulus Determines Lineage Commitment of Cardiac C-kit⁺ Cells

Circulation. 2017 Dec 12;136(24):2359-2372. doi: 10.1161/CIRCULATIONAHA.117.030137. Epub 2017 Oct 11.

Authors

Zhongming Chen^{1

2}, Wuqiang Zhu^{1

3}, Ingrid Bender^{1

2}, Wuming Gong^{1

2

4}, Il-Youp Kwak^{1

2}, Amritha Yellamilli^{2

5}, Thomas J Hodges¹, Natsumi Nemoto^{1

2}, Jianyi Zhang^{1

3}, Daniel J Garry^{1

2

4}, Jop H van Berlo^{6

2

4

5}

Affiliations

¹ Department of Medicine, Cardiovascular Division, University of Minnesota, Minneapolis (Z.C., W.Z., I.B., W.G., I-Y.K., T.J.H., N.N., J.Z., D.J.G., J.H.v.B.).
² Lillehei Heart Institute, University of Minnesota, Minneapolis (Z.C., I.B., W.G., I-Y.K., A.Y., N.N., D.J.G., J.H.v.B.).
³ Department of Biomedical Engineering, University of Alabama at Birmingham (W.Z., J.Z.).
⁴ Stem Cell Institute, University of Minnesota, Minneapolis (W.G., D.J.G., J.H.v.B.).
⁵ Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis (A.Y., J.H.v.B.).
⁶ Department of Medicine, Cardiovascular Division, University of Minnesota, Minneapolis (Z.C., W.Z., I.B., W.G., I-Y.K., T.J.H., N.N., J.Z., D.J.G., J.H.v.B.) jvanberl@umn.edu.

Abstract

Background: Although cardiac c-kit⁺ cells are being tested in clinical trials, the circumstances that determine lineage differentiation of c-kit⁺ cells in vivo are unknown. Recent findings suggest that endogenous cardiac c-kit⁺ cells rarely contribute cardiomyocytes to the adult heart. We assessed whether various pathological stimuli differentially affect the eventual cell fates of c-kit⁺ cells.

Methods: We used single-cell sequencing and genetic lineage tracing of c-kit⁺ cells to determine whether various pathological stimuli would result in different fates of c-kit⁺ cells.

Results: Single-cell sequencing of cardiac CD45^-c-kit⁺ cells showed innate heterogeneity, indicative of the existence of vascular and mesenchymal c-kit⁺ cells in normal hearts. Cardiac pressure overload resulted in a modest increase in c-kit-derived cardiomyocytes, with significant increases in the numbers of endothelial cells and fibroblasts. Doxorubicin-induced acute cardiotoxicity did not increase c-kit-derived endothelial cell fates but instead induced cardiomyocyte differentiation. Mechanistically, doxorubicin-induced DNA damage in c-kit⁺ cells resulted in expression of p53. Inhibition of p53 blocked cardiomyocyte differentiation in response to doxorubicin, whereas stabilization of p53 was sufficient to increase c-kit-derived cardiomyocyte differentiation.

Conclusions: These results demonstrate that different pathological stimuli induce different cell fates of c-kit⁺ cells in vivo. Although the overall rate of cardiomyocyte formation from c-kit⁺ cells is still below clinically relevant levels, we show that p53 is central to the ability of c-kit⁺ cells to adopt cardiomyocyte fates, which could lead to the development of strategies to preferentially generate cardiomyocytes from c-kit⁺ cells.

Keywords: adult stem cells; anthracyclines; c-kit; heart failure; regeneration.

MeSH terms

Animals
Cell Differentiation
Cell Lineage
Cells, Cultured
Doxorubicin / toxicity
Endothelial Cells / physiology*
Mesenchymal Stem Cells / physiology*
Mice
Mice, Inbred C57BL
Mice, Inbred CBA
Mice, Knockout
Myocardium / cytology*
Myocytes, Cardiac / physiology*
Proto-Oncogene Proteins c-kit / metabolism
Sequence Analysis, DNA
Single-Cell Analysis
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism*

Substances

Tumor Suppressor Protein p53
Doxorubicin
Proto-Oncogene Proteins c-kit

Abstract

MeSH terms

Substances

Grants and funding