Myofibroblast Ccn3 is regulated by Yap and Wwtr1 and contributes to adverse cardiac outcomes

Michael A Flinn; Santiago Alvarez-Argote; Makenna C Knas; Victor Alencar Almeida; Samantha J Paddock; Xiaoxu Zhou; Tyler Buddell; Ayana Jamal; Reiauna Taylor; Pengyuan Liu; Jenny Drnevich; Michaela Patterson; Brian A Link; Caitlin C O'Meara

doi:10.3389/fcvm.2023.1142612

Myofibroblast Ccn3 is regulated by Yap and Wwtr1 and contributes to adverse cardiac outcomes

Front Cardiovasc Med. 2023 Mar 14:10:1142612. doi: 10.3389/fcvm.2023.1142612. eCollection 2023.

Authors

Michael A Flinn^{1

2}, Santiago Alvarez-Argote^{1

2}, Makenna C Knas^{1

2}, Victor Alencar Almeida^{1

2}, Samantha J Paddock^{1

2}, Xiaoxu Zhou³, Tyler Buddell^{2

4}, Ayana Jamal^{2

4}, Reiauna Taylor¹, Pengyuan Liu^{1

3}, Jenny Drnevich⁵, Michaela Patterson^{2

4}, Brian A Link^{2

4}, Caitlin C O'Meara^{1

2

6}

Affiliations

¹ Department of Physiology, Medical College of Wisconsin, Milwaukee, WI, United States.
² Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, United States.
³ Institute of Translational Medicine, Zhejiang University School of Medicine, Hangzhou, China.
⁴ Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, WI, United States.
⁵ High Performance Computing in Biology (HPCBio) and the Roy J. Carver Biotechnology Center, University of Illinois, Urbana-Champaign, Champaign, IL, United States.
⁶ Genomics Sciences and Precision Medicine Center, Medical College of Wisconsin, Milwaukee, WI, United States.

Abstract

Introduction: While Yap and Wwtr1 regulate resident cardiac fibroblast to myofibroblast differentiation following cardiac injury, their role specifically in activated myofibroblasts remains unexplored.

Methods: We assessed the pathophysiological and cellular consequence of genetic depletion of Yap alone (Yap ^fl/fl ;Postn ^MCM ) or Yap and Wwtr1 (Yap ^fl/fl ;Wwtr1 ^fl/+ ;Postn ^MCM ) in adult mouse myofibroblasts following myocardial infarction and identify and validate novel downstream factors specifically in cardiac myofibroblasts that mediate pathological remodeling.

Results: Following myocardial infarction, depletion of Yap in myofibroblasts had minimal effect on heart function while depletion of Yap/Wwtr1 resulted in smaller scars, reduced interstitial fibrosis, and improved ejection fraction and fractional shortening. Single cell RNA sequencing of interstitial cardiac cells 7 days post infarction showed suppression of pro-fibrotic genes in fibroblasts derived from Yap ^fl/fl ,Wwtr1 ^fl/+ ;Postn ^MCM hearts. In vivo myofibroblast depletion of Yap/Wwtr1 as well in vitro knockdown of Yap/Wwtr1 dramatically decreased RNA and protein expression of the matricellular factor Ccn3. Administration of recombinant CCN3 to adult mice following myocardial infarction remarkably aggravated cardiac function and scarring. CCN3 administration drove myocardial gene expression of pro-fibrotic genes in infarcted left ventricles implicating CCN3 as a novel driver of cardiac fibrotic processes following myocardial infarction.

Discussion: Yap/Wwtr1 depletion in myofibroblasts attenuates fibrosis and significantly improves cardiac outcomes after myocardial infarction and we identify Ccn3 as a factor downstream of Yap/Wwtr1 that contributes to adverse cardiac remodeling post MI. Myofibroblast expression of Yap, Wwtr1, and Ccn3 could be further explored as potential therapeutic targets for modulating adverse cardiac remodeling post injury.

Keywords: CCN3 (NOV); WWTR1; YAP; adverse remodeling; cardiac fibroblast; hippo pathway; myofibroblast; pathological remodeling.

Abstract

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