MiR-29b and miR-198 overexpression in CD8+ T cells of renal cell carcinoma patients down-modulates JAK3 and MCL-1 leading to immune dysfunction.

Margherita Gigante,Loreto Gesualdo,Giuseppe Lucarelli,Walter J Storkus,Elisabetta Cavalcanti,Maddalena Gigante,Michele Battaglia,Gianluigi Zaza,Paola Pontrelli,Wolfgang Herr,Morena D’Avenia,Elena Ranieri,Matteo Accetturo,Giuseppe Carrieri

doi:10.1186/s12967-016-0841-9

Abstract

BackgroundMammalian microRNAs (miR) regulate the expression of genes relevant for the development of adaptive and innate immunity against cancer. Since T cell dysfunction has previously been reported in patients with renal cell carcinoma (RCC; clear cell type), we aimed to analyze these immune cells for genetic and protein differences when compared to normal donor T cells freshly after isolation and 35 days after in vitro stimulation (IVS) with HLA-matched RCC tumor cells.MethodsWe investigated gene expression profiles of tumor-reactive CD8+ T cells obtained from RCC patient and compared with their HLA-matched healthy sibling donors using a microarray approach. In addition, miRNAs analysis was performed in a validation cohort of peripheral blood CD8+ T cells from 25 RCC patients compared to 15 healthy volunteers.ResultsWe observed that CD8+ T cells from RCC patients expressed reduced levels of anti-apoptotic and proliferation-associated gene products when compared with normal donor T cells both pre- and post-IVS. In particular, JAK3 and MCL-1 were down-regulated in patient CD8+ T cells versus their normal counterparts, likely due to defective suppressor activity of miR-29b and miR-198 in RCC CD8+ T cells. Indeed, specific inhibition of miR-29b or miR-198 in peripheral blood mononuclear cells (PBMCs) isolated from RCC patients, resulted in the up-regulation of JAK3 and MCL-1 proteins and significant improvement of cell survival in vitro.ConclusionsOur results suggest that miR-29b and miR-198 dysregulation in RCC patient CD8+ T cells is associated with dysfunctional immunity and foreshadow the development of miR-targeted therapeutics to correct such T cell defects in vivo.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-016-0841-9) contains supplementary material, which is available to authorized users.

Highlights

Mammalian microRNAs regulate the expression of genes relevant for the development of adaptive and innate immunity against cancer
JAK3 plays an essential role in development, differentiation, proliferation and survival of T-cells [20] and, as we have previously reported, JAK3 mutations are associated with metastatic spread and poor survival of renal cell carcinoma (RCC) patients [21]
Tumor‐specific CD8+ T cell generation from RCC patient TC and allogenic HLA‐matched healthy donors Tissue specimens were obtained from 30 patients undergoing radical or partial nephrectomy for unilateral renal cell carcinoma and primary tumor cells were established in vitro

Summary

Introduction

Mammalian microRNAs (miR) regulate the expression of genes relevant for the development of adaptive and innate immunity against cancer. Since T cell dysfunction has previously been reported in patients with renal cell carcinoma (RCC; clear cell type), we aimed to analyze these immune cells for genetic and protein differences when compared to normal donor T cells freshly after isolation and 35 days after in vitro stimulation (IVS) with HLAmatched RCC tumor cells. We determined that JAK3 and MCL-1 expression is dysregulated in patient versus normal donor CD8+ T cells. JAK3 plays an essential role in development, differentiation, proliferation and survival of T-cells [20] and, as we have previously reported, JAK3 mutations are associated with metastatic spread and poor survival of RCC patients [21]

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