Cardiac c-Kit biology revealed by inducible transgenesis

Natalie A. Gude, Fareheh Firouzi, Kathleen M. Broughton, Kelli Ilves, Kristine P. Nguyen, Christina R. Payne, Veronica Sacchi, Megan M. Monsanto, Alexandria R. Casillas, Farid G. Khalafalla, Bingyan J. Wang, David E. Ebeid, Roberto Alvarez, Walter P. Dembitsky, Barbara A. Bailey, Jop van Berlo, Mark A. Sussman

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Rationale: Biological signifcance of c-Kit as a cardiac stem cell marker and role(s) of c-Kit+ cells in myocardial development or response to pathological injury remain unresolved because of varied and discrepant fndings. Alternative experimental models are required to contextualize and reconcile discordant published observations of cardiac c-Kit myocardial biology and provide meaningful insights regarding clinical relevance of c-Kit signaling for translational cell therapy. Objective: The main objectives of this study are as follows: demonstrating c-Kit myocardial biology through combined studies of both human and murine cardiac cells; advancing understanding of c-Kit myocardial biology through creation and characterization of a novel, inducible transgenic c-Kit reporter mouse model that overcomes limitations inherent to knock-in reporter models; and providing perspective to reconcile disparate viewpoints on c-Kit biology in the myocardium. Methods and Results: In vitro studies confrm a critical role for c-Kit signaling in both cardiomyocytes and cardiac stem cells. Activation of c-Kit receptor promotes cell survival and proliferation in stem cells and cardiomyocytes of either human or murine origin. For creation of the mouse model, the cloned mouse c-Kit promoter drives Histone2BEGFP (enhanced green?uorescent protein; H2BEGFP) expression in a doxycycline-inducible transgenic reporter line. The combination of c-Kit transgenesis coupled to H2BEGFP readout provides sensitive, specifc, inducible, and persistent tracking of c-Kit promoter activation. Tagging effciency for EGFP+/c-Kit+ cells is similar between our transgenic versus a c-Kit knock-in mouse line, but frequency of c-Kit+ cells in cardiac tissue from the knock-in model is 55% lower than that from our transgenic line. The c-Kit transgenic reporter model reveals intimate association of c-Kit expression with adult myocardial biology. Both cardiac stem cells and a subpopulation of cardiomyocytes express c-Kit in uninjured adult heart, upregulating c-Kit expression in response to pathological stress. Conclusions: c-Kit myocardial biology is more complex and varied than previously appreciated or documented, demonstrating validity in multiple points of coexisting yet heretofore seemingly irreconcilable published fndings.

Original languageEnglish (US)
Pages (from-to)57-72
Number of pages16
JournalCirculation research
Volume123
Issue number1
DOIs
StatePublished - Jan 1 2018

Fingerprint

Gene Transfer Techniques
Stem Cells
Cardiac Myocytes
Proto-Oncogene Proteins c-kit
Doxycycline
Cell- and Tissue-Based Therapy
Cell Survival
Myocardium
Theoretical Models
Cell Proliferation
Wounds and Injuries
Proteins

Keywords

  • C-Kit protein
  • Myocardium
  • Myocytes
  • Signal transduction
  • Stem cell
  • cardiac

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

Cite this

Gude, N. A., Firouzi, F., Broughton, K. M., Ilves, K., Nguyen, K. P., Payne, C. R., ... Sussman, M. A. (2018). Cardiac c-Kit biology revealed by inducible transgenesis. Circulation research, 123(1), 57-72. https://doi.org/10.1161/CIRCRESAHA.117.311828

Cardiac c-Kit biology revealed by inducible transgenesis. / Gude, Natalie A.; Firouzi, Fareheh; Broughton, Kathleen M.; Ilves, Kelli; Nguyen, Kristine P.; Payne, Christina R.; Sacchi, Veronica; Monsanto, Megan M.; Casillas, Alexandria R.; Khalafalla, Farid G.; Wang, Bingyan J.; Ebeid, David E.; Alvarez, Roberto; Dembitsky, Walter P.; Bailey, Barbara A.; van Berlo, Jop; Sussman, Mark A.

In: Circulation research, Vol. 123, No. 1, 01.01.2018, p. 57-72.

Research output: Contribution to journalArticle

Gude, NA, Firouzi, F, Broughton, KM, Ilves, K, Nguyen, KP, Payne, CR, Sacchi, V, Monsanto, MM, Casillas, AR, Khalafalla, FG, Wang, BJ, Ebeid, DE, Alvarez, R, Dembitsky, WP, Bailey, BA, van Berlo, J & Sussman, MA 2018, 'Cardiac c-Kit biology revealed by inducible transgenesis', Circulation research, vol. 123, no. 1, pp. 57-72. https://doi.org/10.1161/CIRCRESAHA.117.311828
Gude NA, Firouzi F, Broughton KM, Ilves K, Nguyen KP, Payne CR et al. Cardiac c-Kit biology revealed by inducible transgenesis. Circulation research. 2018 Jan 1;123(1):57-72. https://doi.org/10.1161/CIRCRESAHA.117.311828
Gude, Natalie A. ; Firouzi, Fareheh ; Broughton, Kathleen M. ; Ilves, Kelli ; Nguyen, Kristine P. ; Payne, Christina R. ; Sacchi, Veronica ; Monsanto, Megan M. ; Casillas, Alexandria R. ; Khalafalla, Farid G. ; Wang, Bingyan J. ; Ebeid, David E. ; Alvarez, Roberto ; Dembitsky, Walter P. ; Bailey, Barbara A. ; van Berlo, Jop ; Sussman, Mark A. / Cardiac c-Kit biology revealed by inducible transgenesis. In: Circulation research. 2018 ; Vol. 123, No. 1. pp. 57-72.
@article{f7279807d325453284714d145704b6be,
title = "Cardiac c-Kit biology revealed by inducible transgenesis",
abstract = "Rationale: Biological signifcance of c-Kit as a cardiac stem cell marker and role(s) of c-Kit+ cells in myocardial development or response to pathological injury remain unresolved because of varied and discrepant fndings. Alternative experimental models are required to contextualize and reconcile discordant published observations of cardiac c-Kit myocardial biology and provide meaningful insights regarding clinical relevance of c-Kit signaling for translational cell therapy. Objective: The main objectives of this study are as follows: demonstrating c-Kit myocardial biology through combined studies of both human and murine cardiac cells; advancing understanding of c-Kit myocardial biology through creation and characterization of a novel, inducible transgenic c-Kit reporter mouse model that overcomes limitations inherent to knock-in reporter models; and providing perspective to reconcile disparate viewpoints on c-Kit biology in the myocardium. Methods and Results: In vitro studies confrm a critical role for c-Kit signaling in both cardiomyocytes and cardiac stem cells. Activation of c-Kit receptor promotes cell survival and proliferation in stem cells and cardiomyocytes of either human or murine origin. For creation of the mouse model, the cloned mouse c-Kit promoter drives Histone2BEGFP (enhanced green?uorescent protein; H2BEGFP) expression in a doxycycline-inducible transgenic reporter line. The combination of c-Kit transgenesis coupled to H2BEGFP readout provides sensitive, specifc, inducible, and persistent tracking of c-Kit promoter activation. Tagging effciency for EGFP+/c-Kit+ cells is similar between our transgenic versus a c-Kit knock-in mouse line, but frequency of c-Kit+ cells in cardiac tissue from the knock-in model is 55{\%} lower than that from our transgenic line. The c-Kit transgenic reporter model reveals intimate association of c-Kit expression with adult myocardial biology. Both cardiac stem cells and a subpopulation of cardiomyocytes express c-Kit in uninjured adult heart, upregulating c-Kit expression in response to pathological stress. Conclusions: c-Kit myocardial biology is more complex and varied than previously appreciated or documented, demonstrating validity in multiple points of coexisting yet heretofore seemingly irreconcilable published fndings.",
keywords = "C-Kit protein, Myocardium, Myocytes, Signal transduction, Stem cell, cardiac",
author = "Gude, {Natalie A.} and Fareheh Firouzi and Broughton, {Kathleen M.} and Kelli Ilves and Nguyen, {Kristine P.} and Payne, {Christina R.} and Veronica Sacchi and Monsanto, {Megan M.} and Casillas, {Alexandria R.} and Khalafalla, {Farid G.} and Wang, {Bingyan J.} and Ebeid, {David E.} and Roberto Alvarez and Dembitsky, {Walter P.} and Bailey, {Barbara A.} and {van Berlo}, Jop and Sussman, {Mark A.}",
year = "2018",
month = "1",
day = "1",
doi = "10.1161/CIRCRESAHA.117.311828",
language = "English (US)",
volume = "123",
pages = "57--72",
journal = "Circulation Research",
issn = "0009-7330",
publisher = "Lippincott Williams and Wilkins",
number = "1",

}

TY - JOUR

T1 - Cardiac c-Kit biology revealed by inducible transgenesis

AU - Gude, Natalie A.

AU - Firouzi, Fareheh

AU - Broughton, Kathleen M.

AU - Ilves, Kelli

AU - Nguyen, Kristine P.

AU - Payne, Christina R.

AU - Sacchi, Veronica

AU - Monsanto, Megan M.

AU - Casillas, Alexandria R.

AU - Khalafalla, Farid G.

AU - Wang, Bingyan J.

AU - Ebeid, David E.

AU - Alvarez, Roberto

AU - Dembitsky, Walter P.

AU - Bailey, Barbara A.

AU - van Berlo, Jop

AU - Sussman, Mark A.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Rationale: Biological signifcance of c-Kit as a cardiac stem cell marker and role(s) of c-Kit+ cells in myocardial development or response to pathological injury remain unresolved because of varied and discrepant fndings. Alternative experimental models are required to contextualize and reconcile discordant published observations of cardiac c-Kit myocardial biology and provide meaningful insights regarding clinical relevance of c-Kit signaling for translational cell therapy. Objective: The main objectives of this study are as follows: demonstrating c-Kit myocardial biology through combined studies of both human and murine cardiac cells; advancing understanding of c-Kit myocardial biology through creation and characterization of a novel, inducible transgenic c-Kit reporter mouse model that overcomes limitations inherent to knock-in reporter models; and providing perspective to reconcile disparate viewpoints on c-Kit biology in the myocardium. Methods and Results: In vitro studies confrm a critical role for c-Kit signaling in both cardiomyocytes and cardiac stem cells. Activation of c-Kit receptor promotes cell survival and proliferation in stem cells and cardiomyocytes of either human or murine origin. For creation of the mouse model, the cloned mouse c-Kit promoter drives Histone2BEGFP (enhanced green?uorescent protein; H2BEGFP) expression in a doxycycline-inducible transgenic reporter line. The combination of c-Kit transgenesis coupled to H2BEGFP readout provides sensitive, specifc, inducible, and persistent tracking of c-Kit promoter activation. Tagging effciency for EGFP+/c-Kit+ cells is similar between our transgenic versus a c-Kit knock-in mouse line, but frequency of c-Kit+ cells in cardiac tissue from the knock-in model is 55% lower than that from our transgenic line. The c-Kit transgenic reporter model reveals intimate association of c-Kit expression with adult myocardial biology. Both cardiac stem cells and a subpopulation of cardiomyocytes express c-Kit in uninjured adult heart, upregulating c-Kit expression in response to pathological stress. Conclusions: c-Kit myocardial biology is more complex and varied than previously appreciated or documented, demonstrating validity in multiple points of coexisting yet heretofore seemingly irreconcilable published fndings.

AB - Rationale: Biological signifcance of c-Kit as a cardiac stem cell marker and role(s) of c-Kit+ cells in myocardial development or response to pathological injury remain unresolved because of varied and discrepant fndings. Alternative experimental models are required to contextualize and reconcile discordant published observations of cardiac c-Kit myocardial biology and provide meaningful insights regarding clinical relevance of c-Kit signaling for translational cell therapy. Objective: The main objectives of this study are as follows: demonstrating c-Kit myocardial biology through combined studies of both human and murine cardiac cells; advancing understanding of c-Kit myocardial biology through creation and characterization of a novel, inducible transgenic c-Kit reporter mouse model that overcomes limitations inherent to knock-in reporter models; and providing perspective to reconcile disparate viewpoints on c-Kit biology in the myocardium. Methods and Results: In vitro studies confrm a critical role for c-Kit signaling in both cardiomyocytes and cardiac stem cells. Activation of c-Kit receptor promotes cell survival and proliferation in stem cells and cardiomyocytes of either human or murine origin. For creation of the mouse model, the cloned mouse c-Kit promoter drives Histone2BEGFP (enhanced green?uorescent protein; H2BEGFP) expression in a doxycycline-inducible transgenic reporter line. The combination of c-Kit transgenesis coupled to H2BEGFP readout provides sensitive, specifc, inducible, and persistent tracking of c-Kit promoter activation. Tagging effciency for EGFP+/c-Kit+ cells is similar between our transgenic versus a c-Kit knock-in mouse line, but frequency of c-Kit+ cells in cardiac tissue from the knock-in model is 55% lower than that from our transgenic line. The c-Kit transgenic reporter model reveals intimate association of c-Kit expression with adult myocardial biology. Both cardiac stem cells and a subpopulation of cardiomyocytes express c-Kit in uninjured adult heart, upregulating c-Kit expression in response to pathological stress. Conclusions: c-Kit myocardial biology is more complex and varied than previously appreciated or documented, demonstrating validity in multiple points of coexisting yet heretofore seemingly irreconcilable published fndings.

KW - C-Kit protein

KW - Myocardium

KW - Myocytes

KW - Signal transduction

KW - Stem cell

KW - cardiac

UR - http://www.scopus.com/inward/record.url?scp=85050132415&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85050132415&partnerID=8YFLogxK

U2 - 10.1161/CIRCRESAHA.117.311828

DO - 10.1161/CIRCRESAHA.117.311828

M3 - Article

C2 - 29636378

AN - SCOPUS:85050132415

VL - 123

SP - 57

EP - 72

JO - Circulation Research

JF - Circulation Research

SN - 0009-7330

IS - 1

ER -