Enhanced tumor growth inhibition by mesenchymal stem cells derived from iPSCs with targeted integration of interleukin24 into rDNA loci
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Bo Liu1, Fei Chen1, Yong Wu1, Xiaolin Wang1, Mai Feng1, Zhuo Li1, Miaojin Zhou1, Yanchi Wang1, Lingqian Wu1, Xionghao Liu1 and Desheng Liang1
1The State Key Laboratory of Medical Genetics and School of Life Sciences, Central South University, Changsha, China
Desheng Liang, email: [email protected]
Xionghao Liu, email: [email protected]
Keywords: rDNA locus, gene targeting, MSCs derived from iPSCs, IL24, anti-tumor
Received: August 17, 2016 Accepted: March 13, 2017 Published: March 28, 2017
Induced pluripotent stem cells (iPSCs) are a promising source of mesenchymal stem cells (MSCs) for clinical applications. In this study, we transformed human iPSCs using a non-viral vector carrying the IL24 transgene pHrn-IL24. PCR and southern blotting confirmed IL24 integration into the rDNA loci in four of 68 iPSC clones. We then differentiated a high expressing IL24-iPSC clone into MSCs (IL24-iMSCs) that showed higher expression of IL24 in culture supernatants and in cell lysates than control iMSCs. IL24-iMSCs efficiently differentiated into osteoblasts, chondrocytes and adipocytes. Functionally, IL24-iMSCs induced in vitro apoptosis in B16-F10 melanoma cells more efficiently than control iMSCs when co-cultured in Transwell assays. In vivo tumor xenograft studies in mice demonstrated that IL24-iMSCs inhibited melanoma growth more than control iMSCs did. Immunofluorescence and histochemical analysis showed larger necrotic areas and cell nuclear aggregation in tumors with IL24-iMSCs than control iMSCs, indicating that IL24-iMSCs inhibited tumor growth by inducing apoptosis. These findings demonstrate efficient transformation of iPSCs through gene targeting with non-viral vectors into a rDNA locus. The ability of these genetically modified MSCs to inhibit in vivo melanoma growth is suggestive of the clinical potential of autologous cell therapy in cancer.
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