The CUL5 E3 Ligase Complex Negatively Regulates Central Signaling Pathways in CD8 T Cells

Overview

Journal Nat Commun

Specialty Biology

Date 2024 Jan 19

PMID 38242867

Authors

Xiaofeng Liao

Wenxue Li

Hongyue Zhou

Barani Kumar Rajendran

Ao Li

Jingjing Ren

Yi Luan

David A Calderwood

Benjamin Turk

Wenwen Tang

Yansheng Liu

Dianqing Wu

Affiliations

Soon will be listed here.

Abstract

CD8 T cells play an important role in anti-tumor immunity. Better understanding of their regulation could advance cancer immunotherapies. Here we identify, via stepwise CRISPR-based screening, that CUL5 is a negative regulator of the core signaling pathways of CD8 T cells. Knocking out CUL5 in mouse CD8 T cells significantly improves their tumor growth inhibiting ability, with significant proteomic alterations that broadly enhance TCR and cytokine signaling and their effector functions. Chemical inhibition of neddylation required by CUL5 activation, also enhances CD8 effector activities with CUL5 validated as a major target. Mechanistically, CUL5, which is upregulated by TCR stimulation, interacts with the SOCS-box-containing protein PCMTD2 and inhibits TCR and IL2 signaling. Additionally, CTLA4 is markedly upregulated by CUL5 knockout, and its inactivation further enhances the anti-tumor effect of CUL5 KO. These results together reveal a negative regulatory mechanism for CD8 T cells and have strong translational implications in cancer immunotherapy.

Citing Articles

A genome-wide association study of European advanced cancer patients treated with opioids identifies regulatory variants on chromosome 20 associated with pain intensity.

Minnai F, Shkodra M, Noci S, Esposito M, Brunelli C, Pigni A Eur J Pain. 2024; 29(1):e4764.

PMID: 39629963 PMC: 11616469. DOI: 10.1002/ejp.4764.

Cellular behavior analysis from live-cell imaging of TCR T cell-cancer cell interactions.

Verma A, Yu C, Bachl S, Lopez I, Schwartz M, Moen E bioRxiv. 2024; .

PMID: 39605616 PMC: 11601648. DOI: 10.1101/2024.11.19.624390.

Ubiquitin modification in the regulation of tumor immunotherapy resistance mechanisms and potential therapeutic targets.

Hong Z, Liu F, Zhang Z Exp Hematol Oncol. 2024; 13(1):91.

PMID: 39223632 PMC: 11367865. DOI: 10.1186/s40164-024-00552-0.

CRISPR-Cas gene knockouts to optimize engineered T cells for cancer immunotherapy.

De Castro V, Galaine J, Loyon R, Godet Y Cancer Gene Ther. 2024; 31(8):1124-1134.

PMID: 38609574 DOI: 10.1038/s41417-024-00771-x.

References

Chen Z, Arai E, Khan O, Zhang Z, Ngiow S, He Y . In vivo CD8 T cell CRISPR screening reveals control by Fli1 in infection and cancer. Cell. 2021; 184(5):1262-1280.e22. PMC: 8054351. DOI: 10.1016/j.cell.2021.02.019. View

Jiang T, Zhou C, Ren S . Role of IL-2 in cancer immunotherapy. Oncoimmunology. 2016; 5(6):e1163462. PMC: 4938354. DOI: 10.1080/2162402X.2016.1163462. View

Mehnert M, Li W, Wu C, Salovska B, Liu Y . Combining Rapid Data Independent Acquisition and CRISPR Gene Deletion for Studying Potential Protein Functions: A Case of HMGN1. Proteomics. 2019; 19(13):e1800438. DOI: 10.1002/pmic.201800438. View

Bruderer R, Bernhardt O, Gandhi T, Xuan Y, Sondermann J, Schmidt M . Optimization of Experimental Parameters in Data-Independent Mass Spectrometry Significantly Increases Depth and Reproducibility of Results. Mol Cell Proteomics. 2017; 16(12):2296-2309. PMC: 5724188. DOI: 10.1074/mcp.RA117.000314. View

Gai W, Peng Z, Liu C, Zhang L, Jiang H . Advances in Cancer Treatment by Targeting the Neddylation Pathway. Front Cell Dev Biol. 2021; 9:653882. PMC: 8060460. DOI: 10.3389/fcell.2021.653882. View

Singh M, Ni M, Sullivan J, Hamerman J, Campbell D . B cell adaptor for PI3-kinase (BCAP) modulates CD8 effector and memory T cell differentiation. J Exp Med. 2018; 215(9):2429-2443. PMC: 6122975. DOI: 10.1084/jem.20171820. View

David C, Massague J . Contextual determinants of TGFβ action in development, immunity and cancer. Nat Rev Mol Cell Biol. 2018; 19(7):419-435. PMC: 7457231. DOI: 10.1038/s41580-018-0007-0. View

Xiao Q, Wu J, Wang W, Chen S, Zheng Y, Yu X . DKK2 imparts tumor immunity evasion through β-catenin-independent suppression of cytotoxic immune-cell activation. Nat Med. 2018; 24(3):262-270. PMC: 5840007. DOI: 10.1038/nm.4496. View

Liu Y, Mi Y, Mueller T, Kreibich S, Williams E, van Drogen A . Multi-omic measurements of heterogeneity in HeLa cells across laboratories. Nat Biotechnol. 2019; 37(3):314-322. DOI: 10.1038/s41587-019-0037-y. View

10.

Gattinoni L, Finkelstein S, Klebanoff C, Antony P, Palmer D, Spiess P . Removal of homeostatic cytokine sinks by lymphodepletion enhances the efficacy of adoptively transferred tumor-specific CD8+ T cells. J Exp Med. 2005; 202(7):907-12. PMC: 1397916. DOI: 10.1084/jem.20050732. View

11.

Kumar B, Field N, Kim D, Dar A, Chen Y, Suresh A . The ubiquitin ligase Cul5 regulates CD4 T cell fate choice and allergic inflammation. Nat Commun. 2022; 13(1):2786. PMC: 9120070. DOI: 10.1038/s41467-022-30437-x. View

12.

Chen J, Lopez-Moyado I, Seo H, Lio C, Hempleman L, Sekiya T . NR4A transcription factors limit CAR T cell function in solid tumours. Nature. 2019; 567(7749):530-534. PMC: 6546093. DOI: 10.1038/s41586-019-0985-x. View

13.

Seo H, Chen J, Gonzalez-Avalos E, Samaniego-Castruita D, Das A, Wang Y . TOX and TOX2 transcription factors cooperate with NR4A transcription factors to impose CD8 T cell exhaustion. Proc Natl Acad Sci U S A. 2019; 116(25):12410-12415. PMC: 6589758. DOI: 10.1073/pnas.1905675116. View

14.

Alotaibi F, Rytelewski M, Figueredo R, Zareardalan R, Zhang M, Ferguson P . CD5 blockade enhances ex vivo CD8 T cell activation and tumour cell cytotoxicity. Eur J Immunol. 2020; 50(5):695-704. DOI: 10.1002/eji.201948309. View

15.

Porter D, Levine B, Kalos M, Bagg A, June C . Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia. N Engl J Med. 2011; 365(8):725-33. PMC: 3387277. DOI: 10.1056/NEJMoa1103849. View

16.

Rosenberg S, Packard B, Aebersold P, Solomon D, Topalian S, TOY S . Use of tumor-infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. A preliminary report. N Engl J Med. 1988; 319(25):1676-80. DOI: 10.1056/NEJM198812223192527. View

17.

Pilipow K, Roberto A, Roederer M, Waldmann T, Mavilio D, Lugli E . IL15 and T-cell Stemness in T-cell-Based Cancer Immunotherapy. Cancer Res. 2015; 75(24):5187-5193. PMC: 4681597. DOI: 10.1158/0008-5472.CAN-15-1498. View

18.

Zhang N, Bevan M . Transforming growth factor-β signaling controls the formation and maintenance of gut-resident memory T cells by regulating migration and retention. Immunity. 2013; 39(4):687-96. PMC: 3805703. DOI: 10.1016/j.immuni.2013.08.019. View

19.

Enchev R, Schulman B, Peter M . Protein neddylation: beyond cullin-RING ligases. Nat Rev Mol Cell Biol. 2014; 16(1):30-44. PMC: 5131867. DOI: 10.1038/nrm3919. View

20.

Connolly K, Kuchroo M, Venkat A, Khatun A, Wang J, William I . A reservoir of stem-like CD8 T cells in the tumor-draining lymph node preserves the ongoing antitumor immune response. Sci Immunol. 2021; 6(64):eabg7836. PMC: 8593910. DOI: 10.1126/sciimmunol.abg7836. View