Discovering and Characterizing of Survivin Dominant Negative Mutants With Stronger Pro-apoptotic Activity on Cancer Cells and CSCs

Wei Guo; Xingyuan Ma; Yunhui Fu; Chang Liu; Qiuli Liu; Fabiao Hu; Hui Miao; Tong Zhang; Yuping Liu; Myong Hun Han; Fang You; Yi Yang; Wenyun Zheng

doi:10.3389/fonc.2021.635233

Discovering and Characterizing of Survivin Dominant Negative Mutants With Stronger Pro-apoptotic Activity on Cancer Cells and CSCs

Front Oncol. 2021 Mar 31:11:635233. doi: 10.3389/fonc.2021.635233. eCollection 2021.

Authors

Wei Guo¹, Xingyuan Ma¹, Yunhui Fu¹, Chang Liu¹, Qiuli Liu², Fabiao Hu¹, Hui Miao¹, Tong Zhang², Yuping Liu², Myong Hun Han¹, Fang You³, Yi Yang^{3

4}, Wenyun Zheng²

Affiliations

¹ State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
² Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, China.
³ Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore, Singapore.
⁴ SinGENE Biotech Pte Ltd, Singapore Science Park, Singapore, Singapore.

Abstract

Survivin as a member of the inhibitor of apoptosis proteins (IAPs) family is undetectable in normal cells, but highly expressed in cancer cells and cancer stem cells (CSCs) which makes it an attractive target in cancer therapy. Survivin dominant negative mutants have been reported as competitive inhibitors of endogenous survivin protein in cancer cells. However, there is a lack of systematic comparative studies on which mutants have stronger effect on promoting apoptosis in cancer cells, which will hinder the development of novel anti-cancer drugs. Here, based on the previous study of survivin and its analysis of the relationship between structure and function, we designed and constructed a series of different amino acid mutants from survivin (TmSm34, TmSm48, TmSm84, TmSm34/48, TmSm34/84, and TmSm34/48/84) fused cell-permeable peptide TATm at the N-terminus, and a dominant negative mutant TmSm34/84 with stronger pro-apoptotic activity was selected and evaluated systematically in vitro. The double-site mutant of survivin (TmSm34/84) showed more robust pro-apoptotic activity against A549 cells than others, and could reverse the resistance of A549 CSCs to adriamycin (ADM) (reversal index up to 7.01) by decreasing the expression levels of survivin, P-gp, and Bcl-2 while increasing cleaved caspase-3 in CSCs. This study indicated the selected survivin dominant negative mutant TmSm34/84 is promising to be an excellent candidate for recombinant anti-cancer protein by promoting apoptosis of cancer cells and their stem cells and sensitizing chemotherapeutic drugs.

Keywords: A549 cells and CSCs; discovering and characterizing; sensitizing chemotherapeutic drugs; stronger pro-apoptotic activity; survivin mutant (TmSm34/84).