Browsing by Author "Gordino, Gisela"
Now showing 1 - 2 of 2
Results Per Page
Sort Options
- MicroRNA‐181a restricts human γδ T cell differentiation by targeting Map3k2 and Notch2Publication . Gordino, Gisela; Costa‐Pereira, Sara; Corredeira, Patrícia; Alves, Patrícia; Costa, Luís; Gomes, Anita Q.; Silva‐Santos, Bruno; Ribot, Julie C.γδ T cells are a conserved population of lymphocytes that contributes to anti-tumor responses through its overt type 1 inflammatory and cytotoxic properties. We have previously shown that human γδ T cells acquire this profile upon stimulation with IL-2 or IL-15, in a differentiation process dependent on MAPK/ERK signaling. Here, we identify microRNA-181a as a key modulator of human γδ T cell differentiation. We observe that miR-181a is highly expressed in patients with prostate cancer and that this pattern is associated with lower expression of NKG2D, a critical mediator of cancer surveillance. Interestingly, miR-181a expression negatively correlates with an activated type 1 effector profile obtained from in vitro differentiated γδ T cells and miR-181a overexpression restricts their levels of NKG2D and TNF-α. Upon in silico analysis, we identify two miR-181a candidate targets, Map3k2 and Notch2, which we validate via overexpression coupled with luciferase assays. These results reveal a novel role for miR-181a as a critical regulator of human γδ T cell differentiation and highlight its potential for manipulation of γδ T cells in next-generation immunotherapies.
- miR-181a negatively regulates human γδ T cell differentiation into anti-tumor effectorsPublication . Gordino, Gisela; Pereira, Sara; Gomes, Anita Q.; silva-Santos, Bruno; Ribot, JulieCytotoxicity and IFN-γ production by human γδ T cells underlie their potent anti-tumor functions. importantly, we have previously shown that ex vivo-isolated γδ thymocytes produced negligible IFN-γ and lacked cytolytic activity against leukemia cells but could acquire those properties upon stimulation with IL-2 or IL-15. This notwithstanding, the role of post-transcriptional mechanisms mediated by miRs in the acquisition of the γδ Th1-like phenotype remains unclear. By resorting to RNA-sequencing techniques, we have identified a discrete repertoire of miRs associated with this process, highlighting miR-181a as a potential regulator of γδ T cell functional differentiation. Strikingly, we have observed that miR-181a expression in γδ T cells could be altered by the presence of anti-inflammatory or pro-inflammatory cytokines, either upregulating or downregulation this miR levels, respectively, thus evidencing his role in modulating pathological responses elicited by γδ T cells. Importantly, in a series of gain-of-function experiments, we verified that miR-181a overexpression significantly impaired NKG2D, TNF-α and IFN-γ expression in the Vδ1+ subpopulation. additionally, the RT-PCR analysis of those samples allowed us to verify a decrease in the mRNA levels of genes linked to the cytotoxic potential of these cells. By using luciferase reporter technology, we have been able to validate Map3k2 and Notch2 as direct targets, through which miR-181a elicits his impairment of the Th1-like phenotype in γδ T cells. These findings may have major implications for the manipulation of γδ T cells in cancer immunotherapy.