SRC family kinase (SFK) inhibitor dasatinib improves the antitumor activity of anti-PD-1 in NSCLC models by inhibiting Treg cell conversion and proliferation

Abstract

IntroductionThe use of immune-checkpoint inhibitors has drastically improved the management of patients with non-small cell lung cancer (NSCLC), but innate and acquired resistances are hurdles needed to be solved. Immunomodulatory drugs that can reinvigorate the immune cytotoxic activity, in combination with antiprogrammed cell death 1 (PD-1) antibody, are a great promise to overcome resistance. We evaluated the impact of the SRC family kinases (SFKs) on NSCLC prognosis, and the immunomodulatory effect of the SFK inhibitor dasatinib, in combination with anti-PD-1, in clinically relevant mouse models of NSCLC.MethodsA cohort of patients from University Clinic of Navarra (n=116) was used to study immune infiltrates by multiplex immunofluorescence (mIF) and YES1 protein expression in tumor samples. Publicly available resources (TCGA, Km Plotter, and CIBERSORT) were used to study patient’s survival based on expression of SFKs and tumor infiltrates. Syngeneic NSCLC mouse models 393P and UNSCC680AJ were used for in vivo drug testing.ResultsAmong the SFK members, YES1 expression showed the highest association with poor prognosis. Patients with high YES1 tumor levels also showed high infiltration of CD4+/FOXP3+ cells (regulatory T cells (Tregs)), suggesting an immunosuppressive phenotype. After testing for YES1 expression in a panel of murine cell lines, 393P and UNSCC680AJ were selected for in vivo studies. In the 393P model, dasatinib+anti-PD-1 treatment resulted in synergistic activity, with 87% tumor regressions and development of immunological memory that impeded tumor growth when mice were rechallenged. In vivo depletion experiments further showed that CD8+ and CD4+ cells are necessary for the therapeutic effect of the combination. The antitumor activity was accompanied by a very significant decrease in the number of Tregs, which was validated by mIF in tumor sections. In the UNSCC680AJ model, the antitumor effects of dasatinib+anti-PD-1 were milder but similar to the 393P model. In in vitro assays, we demonstrated that dasatinib blocks proliferation and transforming growth factor beta-driven conversion of effector CD4+ cells into Tregs through targeting of phospholymphocyte-specific protein tyrosine kinase and downstream effectors pSTAT5 and pSMAD3.ConclusionsYES1 protein expression is associated with increased numbers of Tregs in patients with NSCLC. Dasatinib synergizes with anti-PD-1 to impair tumor growth in NSCLC experimental models. This study provides the preclinical rationale for the combined use of dasatinib and PD-1/programmed death-ligand 1 blockade to improve outcomes of patients with NSCLC.

Machine learning and bioinformatics analysis revealed classification and potential treatment strategy in stage 3–4 NSCLC patients

“…Our study further unraveled that PD-0325901 or RO-4987655 might be more applicable to cluster-1 patients with advanced NSCLC. Common antitumor drugs, including gemcitabine and paclitaxel [ 63 – 65 ], might apply to cluster-2, and dasatinib might be more applicable to patients in cluster-3.…”

Identification of Novel Immune-infiltration Related miRNAs that can Predict the Prognosis of Patients with Hepatocellular Carcinoma

“…Tregs, a subset of CD4+ T cells, are central regulators of tolerance and immunity, stabilizing the suppressive phenotype of the TME [34] . Previous studies reported that the in ltration of Tregs in the TME is indicative of poor OS in various cancer subtypes [35][36][37] . Using machine learning techniques, we found that the enrichment of Tregs in the TME is one of the top predictors of poor prognosis in HCC patients.…”

YES1 as a Therapeutic Target for HER2-Positive Breast Cancer after Trastuzumab and Trastuzumab-Emtansine (T-DM1) Resistance Development

“…Previous studies showed the role of YES1 amplification in T-DM1 [ 22 ], trastuzumab and lapatinib [ 7 ], and neratinib [ 19 ] regimens. Among members of the Src family, YES1 expression showed the highest association with poor outcomes in patients with non-small cell lung cancer [ 23 ]. We also previously reported [ 7 ] that higher YES1 expression correlated with poor outcomes for HER2-positive breast cancer patients, and the inhibition of YES1 appears to be an urgent clinical issue in breast cancer and other malignancies.…”