Endothelial OX40 activation facilitates tumor cell escape from T cell surveillance through S1P/YAP-mediated angiogenesis
Baoyu He, Rou Zhao, Baogui Zhang, Hongli Pan, Jilan Liu, Lunhua Huang, Yingying Wei, Dong Yang, Jing Liang, Mingyi Wang, Mingsheng Zhao, Sen Wang, Fengyun Dong, Junfeng Zhang, Yanhua Zhang, Xu Zhang, Xiao Zhang, Guanjun Dong, Huabao Xiong, Qingli Bie, Bin Zhang
Baoyu He, Rou Zhao, Baogui Zhang, Hongli Pan, Jilan Liu, Lunhua Huang, Yingying Wei, Dong Yang, Jing Liang, Mingyi Wang, Mingsheng Zhao, Sen Wang, Fengyun Dong, Junfeng Zhang, Yanhua Zhang, Xu Zhang, Xiao Zhang, Guanjun Dong, Huabao Xiong, Qingli Bie, Bin Zhang
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Research Article Immunology Oncology

Endothelial OX40 activation facilitates tumor cell escape from T cell surveillance through S1P/YAP-mediated angiogenesis

Abstract

Understanding the complexity of the tumor microenvironment is vital for improving immunotherapy outcomes. Here, we report that the T cell costimulatory molecule OX40 was highly expressed in tumor endothelial cells (ECs) and was negatively associated with the prognosis of patients, which is irrelevant to T cell activation. Analysis of conditional OX40 loss- and gain-of-function transgenic mice showed that OX40 signal in ECs counteracted the antitumor effects produced in T cells by promoting angiogenesis. Mechanistically, leucine-rich repeat–containing GPCR5 (Lgr5+ ) cancer stem cells induced OX40 expression in tumor ECs via EGF/STAT3 signaling. Activated OX40 interacted with Spns lysolipid transporter 2 (Spns2), obstructing the export of sphingosine 1-phosphate (S1P) and resulting in S1P intracellular accumulation. Increased S1P directly bound to Yes 1–associated protein (YAP), disrupting its interaction with large tumor suppressor kinase 1 (LATS1) and promoting YAP nuclear translocation. Finally, the YAP inhibitor verteporfin enhanced the antitumor effects of the OX40 agonist. Together, these findings reveal an unexpected protumor role of OX40 in ECs, highlighting the effect of nonimmune cell compartments on immunotherapy.

Authors

Baoyu He, Rou Zhao, Baogui Zhang, Hongli Pan, Jilan Liu, Lunhua Huang, Yingying Wei, Dong Yang, Jing Liang, Mingyi Wang, Mingsheng Zhao, Sen Wang, Fengyun Dong, Junfeng Zhang, Yanhua Zhang, Xu Zhang, Xiao Zhang, Guanjun Dong, Huabao Xiong, Qingli Bie, Bin Zhang

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Figure 7

S1P disrupts the interaction between YAP and the p-LATS1 kinase.

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S1P disrupts the interaction between YAP and the p-LATS1 kinase. (A) HUV...
(A) HUVECs treated with DMSO or S1P (10 μM) were immunostained with an anti-YAP antibody. Scale bars: 5 μm. (B) Expression of YAP downstream genes was evaluated in HUVECs treated with DMSO or S1P using qRT-PCR (n = 3). (C) Subcutaneous tumor models were established using MC38 cells in BALB/c nude mice. The mice were treated with S1P (5 mg/kg, i.p.) or a combination of S1P and verteporfin (n = 8). (D) DMSO- or S1P-treated HUVECs were immunoprecipitated using an anti-YAP antibody followed by blotting with the indicated antibodies. (E) Docking models of YAP–p-LATS1 and YAP-S1P. (F) HUVECs were grown to 10%, 30%, 60%, or 90% confluence. Cell lysates were immunoprecipitated using an anti-YAP antibody. p-LATS1 expression was measured using Western blot analysis in total cell lysates. Metabolites in the immunocomplex were extracted. The quantitative abundance of S1P was measured using an LC-MS–based trace-level metabolite detection method (n = 3). (G) HUVECs were grown to 90% confluence. Then, the cells were treated with DMSO or S1P at the indicated concentrations and subjected to immunoprecipitation using IgG or anti-YAP antibodies. Cells at 20% confluence were used as a control. (H) HUVECs were cultured with S1P-free fetal bovine serum. The cells were transfected with scrambled siRNA (siNC) or mixed siRNAs against SPHK1 and SPHK2. Cell lysates were immunoprecipitated with an anti-YAP antibody and blotted with the indicated antibodies. (I) A diagram summarizing the proposed model in which OX40 activation or accumulated S1P disrupts the YAP and p-LATS1 interaction, leading to increased YAP stability, augmented YAP-TEAD4 interaction, and, ultimately, transactive capacity. Two-tailed Student’s t test (B) or 2-way ANOVA (C) was used for statistical analysis.