Dual Vascular Endothelial Growth Factor Receptor and Fibroblast Growth Factor Receptor Inhibition Elicits Antitumor Immunity and Enhances Programmed Cell Death-1 Checkpoint Blockade in Hepatocellular Carcinoma
Dual Vascular Endothelial Growth Factor Receptor and Fibroblast Growth Factor Receptor Inhibition Elicits Antitumor Immunity and Enhances Programmed Cell Death-1 Checkpoint Blockade in Hepatocellular Carcinoma
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双血管内皮生长因子受体和成纤维细胞生长因子受体抑制可引发抗肿瘤免疫并增强肝细胞癌中的程序性细胞死亡 1 检查点阻断
DOI：
10.1159/000505695
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发表时间：
2020-06-01
期刊：
影响因子：
13.8
通讯作者：
Zhao, Ming
中科院分区：
文献类型：
--
作者：
Deng, Haijing;Kan, Anna;Zhao, Ming
背景与目的：联合抗血管生成治疗、免疫检查点阻断和抗程序性细胞死亡-1(PD-1)抗体治疗肝细胞癌是一种有前途的治疗方法。酪氨酸激酶抑制剂是众所周知的抗血管生成药物，具有与抗PD-1抗体结合的潜力。这项研究探讨了联合治疗可能的潜在免疫调节机制。方法：取抗PD-1治疗后预后不同的患者的肝细胞癌组织标本进行RNA测序(RNA-seq)。分别用重组碱性成纤维细胞生长因子(BFGF)和血管内皮生长因子A(VEGFA)刺激雷瓦替尼或索拉非尼治疗后的T细胞。用流式细胞仪和乳酸脱氢酶试验检测T细胞功能。体内实验是在小鼠H22和HEPA 1-6肝癌模型上进行的。采用多色流式细胞术检测肿瘤微环境(TME)局部免疫浸润情况。用RNA-seq法检测基因调控。免疫组织化学方法检测微血管密度，免疫印迹法检测PD-1配体(PD-L1)的表达。结果：经抗PD-1治疗后，进展期患者的VEGF和成纤维细胞生长因子的基线表达显著高于病情稳定的患者。VEGFA和bFGF显著上调T细胞表面PD-1、细胞毒性T淋巴细胞相关蛋白-4和TIM-3的表达，抑制干扰素-γ和颗粒酶B的分泌，抑制T细胞的细胞毒作用。这种免疫抑制作用可被lenvatinib逆转，但不能被索拉非尼逆转。此外，在H22小鼠肝癌模型中，Lenvatinib/抗PD-1抗体双重治疗比索拉非尼或成纤维细胞生长因子受体(FGFR)抑制剂(BGJ398)具有更好的抗肿瘤效果。Lenvatinib/anti-PD-1联合治疗还可导致长期免疫记忆的形成，同时协同调节TME并增强T细胞的细胞毒作用。最后，Lenvatinib抑制人脐静脉内皮细胞PD-L1的表达，从而改善T细胞的功能。结论：抑制血管内皮生长因子受体和FGFR可增强抗PD-1抗体的疗效。Lenvatinib/anti-PD-1联合治疗可能通过协同调节TME中的效应T细胞功能和相互调节肿瘤血管正常化来发挥抗肿瘤活性。
Background and Aims: Combining anti-angiogenic therapy with immune checkpoint blockade with anti-programmed cell death-1 (PD-1) antibodies is a promising treatment for hepatocellular carcinoma (HCC). Tyrosine kinase inhibitors are well-known anti-angiogenic agents and offer potential for combination with anti-PD-1 antibodies. This study investigated the possible underlying immunomodulatory mechanisms of combined therapy. Methods: HCC tissue samples for RNA-sequencing (RNA-seq) were obtained from patients with differential prognoses following anti-PD-1 treatment. Recombinant basic fibroblast growth factor (bFGF) and vascular endothelial growth factor A (VEGFA) were used to stimulate T cells following lenvatinib or sorafenib treatment, respectively. T cell function was analyzed by flow cytometry and lactate dehydrogenase assay. In vivo experiments were conducted in murine H22 and Hepa 1–6 competent models of HCC. Local immune infiltration in the tumor microenvironment (TME) was assessed using multicolor flow cytometry. Gene regulation was evaluated by RNA-seq. Microvascular density was measured by immunohistochemistry, and PD-1 ligand (PD-L1) induction was quantified by western blot. Results: The baseline expression of VEGF and fibroblast growth factor (FGF) in patients with progressive disease was significantly higher than in patients achieving stable disease following anti-PD-1 treatment. VEGFA and bFGF significantly upregulated the expression of PD-1, cytotoxic T-lymphocyte-associated protein-4, and Tim-3 on T cells, while inhibiting the secretion of interferon gamma (IFNG) and granzyme B and suppressing T cell cytotoxicity. This immunosuppressive effect was reverted by lenvatinib but not sorafenib. Furthermore, dual lenvatinib/anti-PD-1 antibody therapy led to better antitumor effects than either sorafenib or fibroblast growth factor receptor (FGFR) inhibitor (BGJ398) in H22 murine models of HCC. Combined lenvatinib/anti-PD-1 treatment also led to long-term immune memory formation, while synergistically modulating the TME and enhancing the cytotoxic effect of T cells. Finally, lenvatinib inhibited PD-L1 expression on human umbilical vein endothelial cells, which improved the function of T cells. Conclusions: Inhibition of vascular endothelial growth factor receptor and FGFR augmented the efficacy of anti-PD-1 antibodies. Combined lenvatinib/anti-PD-1 treatment appears to exert antitumor activity by synergistically modulating effector T cell function in the TME and by mutually regulating tumor vessel normalization.