CD40 Cross-Linking Induces Migration of Renal Tumor Cell through Nuclear Factor of Activated T Cells (NFAT) Activation.

Paola Pontrelli,Federica Spadaccino,Marilisa Saldarelli,Michele Battaglia,Giuseppe Stefano Netti,Elena Ranieri,Giuseppe Castellano,Loreto Gesualdo,Maddalena Gigante,Walter J Storkus,Margherita Gigante,Giovanni Stallone,Luigi Balducci

doi:10.3390/ijms22168871

Abstract

CD40 crosslinking plays an important role in regulating cell migration, adhesion and proliferation in renal cell carcinoma (RCC). CD40/CD40L interaction on RCC cells activates different intracellular pathways but the molecular mechanisms leading to cell scattering are not yet clearly defined. Aim of our study was to investigate the main intracellular pathways activated by CD40 ligation and their specific involvement in RCC cell migration. CD40 ligation increased the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun NH (2)-terminal kinase (JNK) and p38 MAPK. Furthermore, CD40 crosslinking activated different transcriptional factors on RCC cell lines: AP-1, NFkB and some members of the Nuclear Factor of Activated T cells (NFAT) family. Interestingly, the specific inhibition of NFAT factors by cyclosporine A, completely blocked RCC cell motility induced by CD40 ligation. In tumor tissue, we observed a higher expression of NFAT factors and in particular an increased activation and nuclear migration of NFATc4 on RCC tumor tissues belonging to patients that developed metastases when compared to those who did not. Moreover, CD40-CD40L interaction induced a cytoskeleton reorganization and increased the expression of integrin β1 on RCC cell lines, and this effect was reversed by cyclosporine A and NFAT inhibition. These data suggest that CD40 ligation induces the activation of different intracellular signaling pathways, in particular the NFATs factors, that could represent a potential therapeutic target in the setting of patients with metastatic RCC.

Highlights

Renal cell carcinoma (RCC) is the most common tumor in the kidney
CD40 Ligation Activates Different Signalling Pathways in RCC Cell Lines To define the intracellular signaling pathways potentially activated by CD40 crosslinking and involved in cell motility, we focused our attention on the pathways involving the family of the mitogen-activated protein kinase (MAPK: Jun NH (2)-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), p38), the nuclear factor AP-1 and the activation of NFκB
To define the intracellular signaling pathways potentially activated by CD40 crosslinking and involved in cell motility, we focused our attention on the pathways involving the family of the mitogen-activated protein kinase (MAPK: JNK, ERK, p38), the nuclear4foafc1-5 tor AP-1 and the activation of NFκB

Summary

Introduction

CD40 is a type I membrane glycoprotein that belongs to the tumor necrosis factor receptor superfamily and represent an important T cell co-stimulatory molecule. The isolation of its ligand (CD40L/CD154), a type II transmembrane protein predominantly expressed in activated T cells, paved the way for the demonstration of a key role for CD40-CD40L interaction in the regulation of innate and adaptive immunity and in the induction of tolerance. The widespread expression of CD40 in carcinomas indicates a possible role for this receptor in the pathogenesis of cancer [4]. The cross-linking of CD40 by CD40 ligand may promote the progression of different tumor types through the induction of cell motility and migration [5,6]. It has been demonstrated that in primary RCC cells [7], CD40 ligation drove proliferation and increased motility [3], the intracellular mechanisms leading to this effect have not been yet well defined

Methods

Results

Conclusion