The Calcitriol/Vitamin D Receptor System Regulates Key Immune Signaling Pathways in Chronic Lymphocytic Leukemia

SIMPLE SUMMARY: Calcitriol, the biologically active form of vitamin D, modulates a plethora of cellular processes following its receptor ligation, namely the vitamin D receptor (VDR). Epidemiological studies have linked low blood levels of vitamin D to adverse disease outcome in several B cell malignancies, including chronic lymphocytic leukemia (CLL), for as yet undetermined reasons. In this study, we sought to obtain deeper biological insight into the role of vitamin D in the pathophysiology of CLL. To this end, we investigated whether the calcitriol/VDR system is functional in CLL and analyzed key signaling pathways that are regulated by calcitriol supplementation, while also exploring the role of microenvironmental signals in the regulation of calcitriol/VDR system. Overall, we provide evidence that the calcitriol/VDR system is functional in CLL, regulating signaling pathways critical for cell survival/proliferation. Although microenvironmental triggers can modulate VDR expression and function, calcitriol appears to act independently, alluding to a potential clinical utility of vitamin D supplementation in CLL. ABSTRACT: It has been proposed that vitamin D may play a role in prevention and treatment of cancer while epidemiological studies have linked vitamin D insufficiency to adverse disease outcomes in various B cell malignancies, including chronic lymphocytic leukemia (CLL). In this study, we sought to obtain deeper biological insight into the role of vitamin D and its receptor (VDR) in the pathophysiology of CLL. To this end, we performed expression analysis of the vitamin D pathway molecules; complemented by RNA-Sequencing analysis in primary CLL cells that were treated in vitro with calcitriol, the biologically active form of vitamin D. In addition, we examined calcitriol effects ex vivo in CLL cells cultured in the presence of microenvironmental signals, namely anti-IgM/CD40L, or co-cultured with the supportive HS-5 cells; and, CLL cells from patients under ibrutinib treatment. Our study reports that the calcitriol/VDR system is functional in CLL regulating signaling pathways critical for cell survival and proliferation, including the TLR and PI3K/AKT pathways. Moreover, calcitriol action is likely independent of the microenvironmental signals in CLL, since it was not significantly affected when combined with anti-IgM/CD40L or in the context of the co-culture system. This finding was also supported by our finding of preserved calcitriol signaling capacity in CLL patients under ibrutinib treatment. Overall, our results indicate a relevant biological role for vitamin D in CLL pathophysiology and allude to the potential clinical utility of vitamin D supplementation in patients with CLL.