Li Peng, Zhu Xinhai, Cao Guangchao, Wu Ruan, Li Ke, Yuan Wenhui, Chen Biyun, Sun Guodong, Xia Xichun, Zhang Hua, Wang Xiao, Yin Zhinan, Lu Ligong, Gao Yunfei
Guangdong Provincial Key Laboratory of Tumor Interventional Diagnosis and Treatment, Zhuhai Institute of Translational Medicine, Zhuhai People's Hospital Affiliated with Jinan University, Jinan University, Zhuhai, Guangdong, China.
The Biomedical Translational Research Institute, Faculty of Medical Science, Jinan University, Guangzhou, Guangdong, China.
J Immunother Cancer. 2022 Mar;10(3). doi: 10.1136/jitc-2021-003477.
Epidemiological surveys have revealed that low serum vitamin D level was correlated with increased risk of tumors. Dysfunctional T cells in patients with tumor are characterized as exhausted with high levels of immune checkpoint receptors (ICRs). However, whether the reduced level of vitamin D in patients with cancer correlates with cytotoxic T-cell exhaustion is unknown.
Periphery blood samples from 172 patients with non-small cell lung cancer (NSCLC) were prospectively collected. Patients with NSCLC received one course of intravenous docetaxel (75 mg/m) followed by treatment with or without rocaltrol at a dose of 0.5-2.0 µg/day for total of 3 weeks. We performed phenotypical and functional analysis of T-cell through flow cytometry. Vitamin D receptor (VDR) knockout and overexpression CD8 and Vδ2 T cells were constructed using Cas9-gRNA targeted and overexpressing approaches to identify 1α,25(OH)D/VDR-mediated transcription regulation for ICRs or antitumor activity in T cells.
We show that serum level of vitamin D is negatively correlated with expression of programmed cell death-1 (PD-1), T-cell immunoreceptor with Ig and ITIM domains (TIGIT), and T-cell immunoglobulin and mucin-domain containing-3 (Tim-3), but positively correlated with CD28 expression on CD8 and Vγ9Vδ2 T cells in patients with NSCLC. 1α,25(OH)D, the active form of vitamin D, promotes the nuclear translocation of VDR, which binds to the promoter region of , , and genes and inhibits their expression. Besides, 1α,25(OH)D pretreatment also promotes the methylation of CpG island in the promoter region of the gene and increases H3K27 acetylation at the promoter region of the gene, which leads to surface PD-1 downregulation and CD28 upregulation, respectively. We further reveal that VDR-mediated Ca influx enhanced expression of Th1 cytokines via T-cell receptor activation. Functionally, 1α,25(OH)D pretreated CD8 T cells or Vγ9Vδ2 T cells showed increased Th1 cytokine production and enhanced antitumor immunity. Finally, oral 1α,25(OH)D could also decrease expression of PD-1, Tim-3, TIGIT and increase expression of CD28, resulting in cytokine production (associated with antitumor immunity) by cytotoxic T cells of patients with NSCLC.
Our findings uncover the pleiotropic effects of 1α,25(OH)D in rescuing the exhausted phenotype of human cytotoxic T cells in patients with tumor and in promoting their antitumor immunity.
ChiCTR2100051135.
流行病学调查显示，血清维生素D水平低与肿瘤风险增加相关。肿瘤患者的功能失调T细胞表现为耗竭，免疫检查点受体（ICR）水平高。然而，癌症患者维生素D水平降低是否与细胞毒性T细胞耗竭相关尚不清楚。
前瞻性收集172例非小细胞肺癌（NSCLC）患者的外周血样本。NSCLC患者接受一个疗程的静脉多西他赛（75mg/m）治疗，随后接受或不接受剂量为0.5 - 2.0μg/天的罗钙全治疗，共3周。我们通过流式细胞术对T细胞进行表型和功能分析。使用Cas9 - gRNA靶向和过表达方法构建维生素D受体（VDR）敲除和过表达的CD8和Vδ2 T细胞，以确定1α,25(OH)D/VDR介导的T细胞中ICR或抗肿瘤活性的转录调控。
我们发现，NSCLC患者血清维生素D水平与程序性细胞死亡蛋白1（PD - 1）、含免疫球蛋白和免疫受体酪氨酸抑制基序结构域的T细胞免疫受体（TIGIT）以及含T细胞免疫球蛋白和粘蛋白结构域3（Tim - 3）的表达呈负相关，但与CD8和Vγ9Vδ2 T细胞上CD28的表达呈正相关。维生素D的活性形式1α,25(OH)D促进VDR的核转位，VDR与 、 和 基因的启动子区域结合并抑制其表达。此外，1α,25(OH)D预处理还促进 基因启动子区域CpG岛的甲基化，并增加 基因启动子区域的组蛋白H3K27乙酰化，分别导致表面PD - 1下调和CD28上调。我们进一步揭示，VDR介导的钙内流通过T细胞受体激活增强了Th1细胞因子的表达。在功能上，1α,25(OH)D预处理的CD8 T细胞或Vγ9Vδ2 T细胞显示Th1细胞因子产生增加和抗肿瘤免疫力增强。最后，口服1α,25(OH)D也可降低PD - 1、Tim - 3、TIGIT的表达并增加CD28的表达，导致NSCLC患者细胞毒性T细胞产生细胞因子（与抗肿瘤免疫相关）。
我们的研究结果揭示了1α,25(OH)D在挽救肿瘤患者人细胞毒性T细胞的耗竭表型和促进其抗肿瘤免疫方面的多效性作用。
ChiCTR2100051135。
