Enhanced tumor growth inhibition by mesenchymal stem cells derived from iPSCs with targeted integration of interleukin24 into rDNA loci

Abstract

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.

Human Wharton's Jelly Stem Cell (hWJSC) Extracts Inhibit Ovarian Cancer Cell Lines OVCAR3 and SKOV3 in vitro by Inducing Cell Cycle Arrest and Apoptosis

“…Mesenchymal stem cells (MSCs) including the hWJSCs derived from within the umbilical cord have been used as anticancer agents either naïve or engineered against various human cancers (20–22). The hWJSCs have many advantages compared to other currently available MSCs such as painless harvest as they are derived from umbilical cord which are usually discarded at birth, relatively young when derived, highly proliferative, have long telomeres and widely multipotent (14).…”

Paired CRISPR/Cas9 Nickases Mediate Efficient Site-Specific Integration of F9 into rDNA Locus of Mouse ESCs

“…These properties indicate that the rDNA locus may be an ideal safe locus for transgene integration [ 7 , 8 ]. In our previous studies, exogenous genes have been targeted to the rDNA region and stably expressed in various cell lines by non-viral plasmids, including immortal adult cells and stem cells [ 9 , 10 , 11 , 12 , 13 ].…”

Inhibition of Colorectal Cancer Cells HT-29 by Secretome and Extraction of Human Umbilical Cord Wharton’s Jelly Stem Cells

“…In this regards, several studies reported that hWJSCs have an appropriate anticancer activity through various mechanisms [5,9]. This stem cells are easily derived from umbilical cord and suggested as an anticancer agent [16,17]. The hWJSCs presents several advantages as compared to other stem cells, and are derived from human umbilical cord which are discarded at birth.…”