Mesenchymal stem cells pretreated with delivered Hph-1-Hsp70 protein are protected from hypoxia-mediated cell death and rescue heart functions from myocardial injury.
Mesenchymal stem cell (MSC) therapy for myocardial injury has inherent limitations due to the poor viability of MSCs after cell transplantation. In this study, we directly delivered Hsp70, a protein with protective functions against stress, into MSCs, using the Hph-1 protein transduction domain ex vivo for high transfection efficiency and low cytotoxicity. Compared to control MSCs in in vitro hypoxic conditions, MSCs delivered with Hph-1-Hsp70 (Hph-1-Hsp70-MSCs) displayed higher viability and anti-apoptotic properties, including Bcl2 increase, reduction of Bax, JNK phosphorylation and caspase-3 activity. Hsp70 delivery also attenuated cellular ATP-depleting stress. Eight animals per group were used for in vivo experiments after occlusion of the left coronary artery. Transplantation of Hph-1-Hsp70-MSCs led to a decrease in the fibrotic heart area, and significantly reduced the apoptotic positive index by 19.5 +/- 2%, compared to no-treatment controls. Hph-1-Hsp70-MSCs were well-integrated into the infarcted host myocardium. The mean microvessel count per field in the infarcted myocardium of the Hph-1-Hsp70-MSC-treated group (122.1 +/- 13.5) increased relative to the MSC-treated group (75.9 +/- 10.4). By echocardiography, transplantation of Hph-1-Hsp70-MSCs resulted in additional increases in heart function, compared to the MSCs-transplanted group. Our results may help formulate better clinical strategies for in vivo MSC cell therapy for myocardial damage.
Chang
Woochul
W
Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut, USA.
Song
Byeong-Wook
BW
Lim
Soyeon
S
Song
Heesang
H
Shim
Chi Young
CY
Cha
Min-Ji
MJ
Ahn
Dong Hyuck
DH
Jung
Young-Gook
YG
Lee
Dong-Ho
DH
Chung
Ji Hyung
JH
Choi
Ki-Doo
KD
Lee
Seung-Kyou
SK
Chung
Namsik
N
Lee
Sang-Kyou
SK
Jang
Yangsoo
Y
Hwang
Ki-Chul
KC
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000235:genetics / Q000502:physiology*; D015687:Cell Hypoxia / Q000235:genetics / Q000502:physiology*; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000502:physiology*; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D009203:Myocardial Infarction / Q000628:therapy*; D063150:Polycomb Repressive Complex 1; D051381:Rats; D017207:Rats, Sprague-Dawley
D002352:Carrier Proteins; D018840:HSP70 Heat-Shock Proteins; C103827:Phc1 protein, mouse; D000255:Adenosine Triphosphate; D063150:Polycomb Repressive Complex 1; D053148:Caspase 3