SHP2 deneddylation mediates tumor immunosuppression in colon cancer via the CD47/SIRPα axis
Yiqing Li, … , Xue Zhang, Yuehai Ke
Yiqing Li, … , Xue Zhang, Yuehai Ke
Published January 10, 2023
Citation Information: J Clin Invest. 2023;133(4):e162870. https://doi.org/10.1172/JCI162870.
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Research Article Cell biology

SHP2 deneddylation mediates tumor immunosuppression in colon cancer via the CD47/SIRPα axis

Abstract

SIPRα on macrophages binds with CD47 to resist proengulfment signals, but how the downstream signal of SIPRα controls tumor-infiltrating macrophages (TIMs) is still poorly clarified. Here, we report that the CD47/signal regulatory protein α (SIRPα) axis requires the deneddylation of tyrosine phosphatase SHP2. Mechanistically, Src homology region 2–containing protein tyrosine phosphatase 2 (SHP2) was constitutively neddylated on K358 and K364 sites; thus, its autoinhibited conformation was maintained. In response to CD47-liganded SIRPα, SHP2 was deneddylated by sentrin-specific protease 8 (SENP8), which led to the dephosphorylation of relevant substrates at the phagocytic cup and subsequent inhibition of macrophage phagocytosis. Furthermore, neddylation inactivated myeloid-SHP2 and greatly boosted the efficacy of colorectal cancer (CRC) immunotherapy. Importantly, we observed that supplementation with SHP2 allosteric inhibitors sensitized immune treatment–resistant CRC to immunotherapy. Our results emphasize that the CRC subtype that is unresponsive to immunotherapy relies on SIRPαhiSHP2hiNEDD8lo TIMs and highlight the need to further explore the strategy of SHP2 targeting in CRC therapy.

Authors

Yiqing Li, Hui Zhou, Pan Liu, Dandan Lv, Yichun Shi, Bufu Tang, Jiaqi Xu, Tingting Zhong, Wangting Xu, Jie Zhang, Jianying Zhou, Kejing Ying, Yongchao Zhao, Yi Sun, Zhinong Jiang, Hongqiang Cheng, Xue Zhang, Yuehai Ke

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Figure 3

SHP2 is a genuine substrate of NEDD8.

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SHP2 is a genuine substrate of NEDD8. (A) Western blot indicating SHP2 n...
(A) Western blot indicating SHP2 neddylation of BMDMs. (B) Western blot indicating NEDD8-SHP2 conjugation relied on the isopeptide bond of HEK293T cells (n = 3). (C) Western blot indicating depletion of SENP8 enhanced SHP2 neddylation of BMDMs (n = 3). (D–F) SHP2 neddylation in BMDMs was attenuated by E1 inhibitor MLN4924 (1 μM, 8 hours) (D) or deletion of UBE2M (F) but not UBE2F (E) (n = 3). (G) Western blot indicating Poly-NEDD8 chain was not involved in SHP2 neddylation of HEK293T cells (n = 3). (H) Western blot indicating sites of SHP2 neddylation of HEK293T cells (n = 3). (I) Western blot indicating domains of SHP2 neddylation of HEK293T cells (n = 3). (J) Docking of molecules between NEDD8 and SHP2. (K) Upper panel: RMSF of SHP2 residents in MD. Lower panel: color overlay of the time-frame configurations in 1–100 amino acid area of SHP2. (L) Western blot indicating the state of SHP2 variant neddylation of HEK293T cells (n = 3). (M) Western blot indicating in vitro neddylation assay (n = 3). Recombinant SHP2 and its indicated mutation (1 μM), E3 XIAP (2.5 μM). D-IP, denaturing IP; WCL, whole-cell lysate. Data are represented as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; nonsignificant (NS), P > 0.05. Two-tailed, unpaired Student’s t test (B, C, D, F); 2-way ANOVA followed by Tukey’s post hoc test (H, L, M);