Adeno-associated virus 9 vector-mediated cardiac-selective expression of human secretory leukocyte protease inhibitor attenuates myocardial ischemia/reperfusion injury

Podsawee Mongkolpathumrat; Nitirut Nernpermpisooth; Anusak Kijtawornrat; Faprathan Pikwong; Wannapat Chouyratchakarn; Rungrueang Yodsheewan; Sasimanas Unajak; Sarawut Kumphune

doi:10.3389/fcvm.2022.976083

Adeno-associated virus 9 vector-mediated cardiac-selective expression of human secretory leukocyte protease inhibitor attenuates myocardial ischemia/reperfusion injury

Front Cardiovasc Med. 2022 Aug 19:9:976083. doi: 10.3389/fcvm.2022.976083. eCollection 2022.

Authors

Podsawee Mongkolpathumrat¹, Nitirut Nernpermpisooth^{2

3}, Anusak Kijtawornrat⁴, Faprathan Pikwong¹, Wannapat Chouyratchakarn¹, Rungrueang Yodsheewan⁵, Sasimanas Unajak⁶, Sarawut Kumphune^{1

2}

Affiliations

¹ Biomedical Engineering Institute (BMEI), Chiang Mai University, Chiang Mai, Thailand.
² Integrative Biomedical Research Unit (IBRU), Faculty of Allied Health Sciences, Naresuan University, Phitsanulok, Thailand.
³ Department of Cardio-Thoracic Technology, Faculty of Allied Health Sciences, Naresuan University, Phitsanulok, Thailand.
⁴ Department of Physiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand.
⁵ Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.
⁶ Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, Thailand.

Abstract

Protease enzymes contribute to the initiation of cardiac remodeling and heart failure after myocardial ischemic/reperfusion (I/R) injury. Protease inhibitors attenuate protease activity and limit left ventricular dysfunction and remodeling. Previous studies showed the cardioprotective effect of secretory leukocyte protease inhibitor (SLPI) against I/R injury. However, overexpression of SLPI gene in cardiovascular diseases has only been investigated in an in vitro experiment. Here, cardiac-selective expression of the human secretory leukocyte protease inhibitor (hSLPI) gene and its effect on I/R injury were investigated. Adeno-associated virus (AAV) serotype 9 carrying hSLPI under the control of cardiac-selective expression promoter (cardiac troponin, cTn) was intravenously administered to Sprague-Dawley rats for 4 weeks prior to coronary artery ligation. The results showed that myocardial-selective expression of hSLPI significantly reduced infarct size, cardiac troponin I (cTnI), creatine kinase-MB (CK-MB), and myoglobin levels that all served to improve cardiac function. Moreover, overexpression of hSLPI showed a reduction in inflammatory cytokines, oxidatively modified protein carbonyl (PC) content, ischemia-modified albumin (IMA), and necrosis and cardiac tissue degeneration. In conclusion, this is the first study to demonstrate cardiac-selective gene delivery of hSLPI providing cardioprotection against myocardial I/R injury in an in vivo model.

Keywords: cardiac selective expression; cardioprotection; gene therapy; ischemic/reperfusion injury; secretory leukocyte protease inhibitor (SLPI).