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 2

Confirmation of high expression of OX40 specifically in tumor ECs and the protumor effects.

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Confirmation of high expression of OX40 specifically in tumor ECs and th...
(A) OX40 gene expression in various cell subpopulations of CRC and NT tissues using scRNA-Seq data (n = 5). FC, fold change. (B) Proportion of OX40– and OX40+ ECs in CRC and NT tissues (n = 5). (C) Representative images of multicolor immunofluorescence using anti-OX40 (green) and anti-CD31 (red) antibodies in CRC and NT tissues (n = 162). Scale bars: 30 μm. (D) Analysis of the OX40+ EC percentages in CRC and NT tissues. The data were obtained from a tissue microarray, which included 162 pairs of CRC and NT tissues. (E) OX40 expression in sorted CD31+ cells from CRC and NT tissues (n = 3). (F) First, 162 CRCs were divided into 2 groups based on the median values of Ki67+ percentages in CD3+ T cells: CD3+Ki67– and CD3+Ki67+. Further, these 2 groups were divided into 4 parts based on the median values of OX40+ percentages in ECs: CD3+Ki67–;CD31+OX40– (n = 41), CD3+Ki67–;CD31+OX40+ (n = 40), CD3+Ki67+;CD31+OX40– (n = 41), and CD3+Ki67+;CD31+OX40+ (n = 40). Kaplan-Meier analysis of the overall survival probability of patients in the 4 groups was performed. (G–J) C57BL/6J mice with conditional knockin (Ox40ki/ki;Cd31cre/–) or knockout (Ox40fl/fl;Cd31cre/–) of OX40 in ECs were established using the CRISPR/Cas9 method. Subcutaneous tumors (G and I) and splenic injection for the liver metastasis model (H and J) were constructed in these mice using MC38 cells and treated with recombinant mouse OX40L (n = 5 or 10). In H and J, the red arrows indicate the implanted primary tumor in the spleen, and the white arrows indicate metastatic tumor lesions. Two-tailed Student’s t test (D and E), log-rank test (F), 2-way ANOVA (G and I), or 1-way ANOVA (H and J) was used for statistical analysis.