Overcoming the UCB HSCs –Derived NK cells Dysfunction through Harnessing RAS/MAPK, IGF-1R and TGF-β Signaling Pathways

Abstract

Background The natural killer (NK) cells differentiated from umbilical cord blood (UCB) hematopoietic stem cells (HSCs) may be more suitable for cell-based immunotherapy compared to the NK cells from adult donors. This is due to the possibility to choose alloreactive donors and potentially more robust in vivo expansion. However, the cytotoxicity of UCB-HSC-derived NK cells against cancer cells might be suboptimal. To overcome this obstacle, we attempted to generate NK cells with potent antitumor activity by targeting RAS/MAPK, IGF-1R and TGF-β signaling pathways using IL-15, IGF-1 and SIS3 respectively. Methods The CD34 + cells were isolated from human UCB mononuclear cells through magnetic activation cell sorting (MACS) with purity of (≥ 90%) and were subjected to differentiate into NK cells. After 21 days of induction with SFTG36 (SCF, FLt-3L, TPO, GM-CSF, IL-3 and IL-6), IS721 (IGF-1, SIS3, IL-7 and IL-21) and IL-15/Hsp70 media, NK cells phenotypes were studied and their cytotoxicity against K562 human erythroleukemia cells and SKOV3 ovarian carcinoma cells was analyzed. Results The NK cells induced in SFTG36/IS721 medium were selected for activation due to their higher expression of CD56 + 16 + CD3 − (93.23% ± 0.75) and mean fluorescence intensity (MFI) of NKG2D + (168.66 ± 20.00) and also a higher fold expansion potential (11.893 ± 1.712) compared to the other groups. These cells once activated with IL-15, demonstrated a higher cytotoxicity against K562 (≥ 90%; P ≤ 0.001) and SKOV3 tumor cells (≥ 65%; P ≤ 0.001) compared to IL-15/Hsp70-activated NK cells. Conclusions The differentiation of ex vivo expanded CD34 + cells through manipulation of RAS/MAPK, IGF-1R and TGF-β signaling pathways is an efficient approach for generating functional NK cells that can be used for cancer immunotherapy.

Role of RAS signaling in ovarian cancer

“…194 A recent study by Shokouhifar et al highlights the protocol for the generation of natural killer cells from umbilical cord blood hematopoietic stem cells by manipulating RAS/MAPK, IGF-1R and TGF-β signaling pathways that can be used for cancer immunotherapy. 195 RAS associated acquisition of chemoresistance in OC is depicted in Figure 3. Though the mechanism underlying chemoresistance in OC is still ambiguous, numerous such reports suggests the integral role of CSCs in chemoresistance and recurrence.…”

Stem Cell Therapy and Innate Lymphoid Cells

“…Since in haplo-HSCT conditioning regimen, infections, or residual leukemia cells may induce IL-1 β production, this may influence the NK/ILC3 development from donor-derived CD34+ cells [ 74 ]. Shokouhifar et al found that the differentiation of ex vivo expanded CD34+ cells through manipulation of RAS/MAPK, IGF-1R, and TGF- β signaling pathways is an efficient approach for generating functional NK cells that can be used for cancer immunotherapy [ 83 ]. CD34 + -HSPC cultured in the absence or in the presence of the EZH1/2 inhibitor UNC1999 and EZH2 inhibitor GSK126 showed that UNC1999 and GSK126 increased CD56 + cell proliferation in comparison to the control.…”

NK cell upraise in the dark world of cancer stem cells

“…To develop NK cell-based therapies, it is important to find appropriate sources of these cells that minimize cell number limitations, immunological complications, and HLA Donor-Recipient matching [ 89 – 91 ]. There are several sources of NK cell including peripheral blood (PB), umbilical cord blood (UCB) [ 92 ], bone marrow (BM) and cell lines [ 93 ]. Deriving NK cells from PBMC, hESC and iPSC sources was a revolution which wiped out the worries about the low number and purity of NK cells forever [ 94 ].…”