Amelioration of experimental tendinopathy by lentiviral CD44 gene therapy targeting senescence-associated secretory phenotypes

Shih-Yao Chen; I-Ming Jou; Po-Yen Ko; Kai-Lan Hsu; Wei-Ren Su; Li-Chieh Kuo; Pei-Yuan Lee; Chao-Liang Wu; Po-Ting Wu

doi:10.1016/j.omtm.2022.06.006

Amelioration of experimental tendinopathy by lentiviral CD44 gene therapy targeting senescence-associated secretory phenotypes

Mol Ther Methods Clin Dev. 2022 Jun 10:26:157-168. doi: 10.1016/j.omtm.2022.06.006. eCollection 2022 Sep 8.

Authors

Shih-Yao Chen¹, I-Ming Jou^{2

3}, Po-Yen Ko^{4

5}, Kai-Lan Hsu^{4

5}, Wei-Ren Su^{4

6}, Li-Chieh Kuo^{7

8}, Pei-Yuan Lee^{9

10}, Chao-Liang Wu^{11

12}, Po-Ting Wu^{4

5

6

8

11}

Affiliations

¹ Department of Nursing, College of Nursing, Chung Hwa University of Medical Technology, Tainan 71703, Taiwan.
² Department of Orthopaedics, E-Da Hospital, Kaohsiung 82445, Taiwan.
³ School of Medicine, College of Medicine, I-Shou University, Kaohsiung 82445, Taiwan.
⁴ Department of Orthopaedics, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan 70403, Taiwan.
⁵ Department of Biomedical Engineering, National Cheng Kung University, Tainan 70101, Taiwan.
⁶ Department of Orthopaedics, College of Medicine, National Cheng Kung University, Tainan 70101, Taiwan.
⁷ Department of Occupational Therapy, National Cheng Kung University, Tainan 70101, Taiwan.
⁸ Medical Device Innovation Center, National Cheng Kung University, Tainan 70101, Taiwan.
⁹ Department of Orthopaedic Surgery, Show Chwan Memorial Hospital, Changhua 50544, Taiwan.
¹⁰ College of Nursing and Health Sciences, Da-Yeh University, Changhua 51500, Taiwan.
¹¹ Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan 70101, Taiwan.
¹² Department of Medical Research, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chiayi 60002, Taiwan.

Abstract

CD44 exerts anti-senescence effects in many disease models. We examined senescence in tendinopathy and the effect of CD44 on senescence-associated secretory phenotypes (SASPs). Senescent markers were determined in human tendinopathic long head of bicep (LHB) and normal hamstring tendons. CD44 gene transfer in rat tendinopathic tenocytes stimulated with interleukin (IL)-1β and a rat Achilles tendinopathy model were performed using lentiviral vectors. Expression levels of p53, p21, and p16 and senescence-associated β-galactosidase (SA-β-gal) activity were positively correlated with the severity of human tendinopathy and were higher in rat and human tendinopathic tenocytes than in normal controls. CD44 overexpressed tenocyte transfectants exhibited reduced levels of IL-6, matrix metalloproteinases (MMPs), cyclooxygenase (COX)-2, p53, p21, p16, SA-β-gal, and phospho-nuclear factor (NF)-κB, whereas their collagen type I alpha 1 (COL1A1) and tenomodulin (tnmd) levels were increased when compared with control transfectants under IL-1β-stimulated conditions. In the animal model, CD44 overexpression lowered the ultrasound and histology scores and expression levels of the senescent and SASP markers COX-2 and phospho-NF-κB. Bromodeoxyuridine (BrdU)- and tnmd-positive cell numbers were increased in the LVCD44-transduced tendinopathic tendons. Senescence is positively correlated with tendinopathic severity, and CD44 overexpression may protect the tendinopathic tendons from SASPs via anti-inflammation and maintenance of extracellular matrix homeostasis.

Keywords: CD44; gene therapy; lentiviral vector; senescence-associated secretory phenotypes; tendinopathy.