IL2RA x PD-1 x VEGF

Melanoma is an aggressive and highly metastatic malignancy with significant mortality. Although immune checkpoint inhibitors have improved outcomes in advanced disease, immune resistance remains a critical barrier to durable responses, emphasizing the need to define the molecular mechanisms underlying therapeutic failure. Interleukin (IL)30 has been identified as a cancer progression driver with immunosuppressive function in the tumour-microenvironment. Whether it is involved in melanoma aggressiveness and metastasis is unknown. Immunopathological studies, RNA-Seq and CIBERSORTx analyses of tissue samples from the TCGA-SKCM patient cohort, reveal that IL30 expression is absent in normal skin, while it can be found in melanoma and infiltrating immune cells, mostly macrophages. The putative IL30 receptor complex, IL-6Rα/gp130, is expressed by melanoma cells, which respond to IL30 stimulation with increased proliferation and migration, consistent with IL30-mediated upregulation of metastasis-associated genes, including ANG2, CXCR4, ITGB1, MMP2, NME, SNAI2, VEGFA, VEGFC, and L1CAM. Bioinformatic analysis of IL30 transcript levels in TCGA-SKCM tissue specimens, reveals increased IL30 expression in metastases compared with primary tumors, corroborating the experimental findings at the translational level. IL30 reprograms the melanoma immunophenotype by inducing multiple immune checkpoint molecules, including LSECtin, LGALS3, LGALS9, LAG-3, TIM-3, B7-H4, B7-H3, VISTA, and PD-1. Concurrently, IL30 suppresses T cell function by reducing CD25 and HLA-DR expression on CD4⁺ and CD8⁺ T cells, inhibiting their activation and proliferation, decreasing TNF-α and IFN-γ production, and boosting LAG-3 expression, which strongly correlates with IL30 levels in clinical samples. Collectively, these findings identify IL30 as a critical driver of melanoma dissemination and T cell exhaustion, providing a mechanistic link to immune resistance and failure of combination immunotherapies.

Immunotherapy is being tested in early‐stage non‐small cell lung cancer (NSCLC), and achieving higher rates of complete pathological responses (CPR) as compared to standard of care. Early identification of CPR patients has vital clinical implications. In this study, we focused on basal peripheral immune cells and their treatment‐related changes to find biomarkers associated to CPR.

Blood from 29 stage IIIA NSCLC patients participating in the NADIM trial (NCT03081689) was collected at diagnosis and post neoadjuvant treatment. More than 400 parameters of peripheral blood mononuclear cells (PBMCs) phenotype and plasma soluble factors were analyzed.

Neoadjuvant chemoimmunotherapy altered more than 150 immune parameters. At diagnosis, 11 biomarkers associated to CPR were described, with an area under the ROC curve >0.70 and p ‐value <.05. CPR patients had significantly higher levels of CD4 + PD‐1 + cells, NKG2D, and CD56 expression on T CD56 cells, intensity of CD25 expression on CD4 + CD25hi + cells and CD69 expression on intermediate monocytes; but lower levels of CD3 + CD56 – CTLA‐4 + cells, CD14 ++ CD16 + CTLA‐4 + cells, CTLA‐4 expression on T CD56 cells and lower levels of b‐NGF, NT‐3, and VEGF‐D in plasma compared to non‐CPR. Post treatment, CPR patients had significantly higher levels of CD19 expression on B cells, BCMA, 4‐1BB, MCSF, and PARC and lower levels of MPIF‐1 and Flt‐3L in plasma compared to non‐CPR.

Patients achieving CPR seem to have a distinctive peripheral blood immune status at diagnosis, even showing different immune response to treatment. These results reinforce the different biology behind CPR and non‐CPR responses.

The study was designed to optimize the condition for producing an effective dendritic cell (DCs) based immunotherapy by using DCs and T lymphocytes together with tumor-infiltrating lymphocytes (TILs) and tumor-infiltrating DCs (TIDCs), treated with anti-PD1 and anti-CTLA4 monoclonal antibodies. This mixture of immune cells was co-cultured with autologous breast cancer cells (BCCs) isolated from 26 BC females.

There was a significant upregulation of CD86 and CD83 on DCs (p = 0.001 and 0.017, respectively), similarly upregulation of CD8, CD4 and CD103 on T cells (p = 0.031, 0.027, and 0.011, respectively). While there was a significant downregulation of FOXP3 and combined CD25.CD8 expression on regulatory T cells (p = 0.014 for both). Increased CD8/Foxp3 ratio (p < 0.001) was also observed. CD133, CD34 and CD44 were downregulated on BCCs (p = 0.01, 0.021, and 0.015, respectively). There was a significant increase in interferon-γ (IFN-γ, p < 0.001), lactate dehydrogenase (LDH, p = 0.02), and a significant decrease in vascular endothelial growth factor (VEGF, p < 0.001) protein levels. Gene expression of FOXP3 and Programmed cell death ligand 1 (PDL-1) were downregulated in BCCs (p < 0.001, for both), similarly cytotoxic T lymphocyte antigen-4 (CTLA4, p = 0.02), Programmed cell death 1 (PD-1, p < 0.001) and FOXP3 (p < 0.001) were significantly downregulated in T cells.

Ex-vivo activation of immune cells (DCs, T cells, TIDCs, and TILs) with immune checkpoint inhibitors could produce a potent and effective BC immunotherapy. However, these data should be validated on an experimental animal model to be transferred to the clinical setting.