Tumor-infiltrating TNFRSF9⁺ CD8⁺ T cells define different subsets of clear cell renal cell carcinoma with prognosis and immunotherapeutic response

ABSTRACT

Objectives

Tumor necrosis receptor super family (TNFRSF) plays an important role in regulating the function of CD8⁺ T cells. In this study, we explored the clinical significance and immune profile of TNFRSF9⁺ CD8⁺ T cells in clear cell renal cell carcinoma (ccRCC)

Methods

The infiltration of immune cells was determined by immunohistochemistry in ZS cohort from our hospital and their prognostic value was further determined by Cox regression. Functional status of CD8⁺ T cells in ccRCC was determined by flow cytometry in 29 fresh tumor samples. In silico analysis on a TCGA cohort and other datasets was performed to further demonstrate our findings.

Results

High TNFRSF9⁺ CD8⁺ T cells infiltration was associated with inferior overall survival in ZS cohort (p = .0016) and TCGA-KIRC cohort (p = .018). TNFRSF9⁺ CD8⁺ T cells expressed higher exhaustion markers (PD-1, TIM-3, CTLA-4, and TIGIT), and effector markers (IFN-γ, GZMB, CD107a, and Ki-67), than their TNFRSF9 negative counterparts. In silico analysis indicated the expression of TNFRSF9 was significantly correlated with IFNG, GZMK, MKI-67, PDCD1, HAVCR2, TIGIT, and CTLA-4 in CD8⁺ T cells. However, higher TNFRSF9 signature was correlated with larger tumor size shrinkage (p = .003) and better progression-free survival (p = .012) in patients treated with nivolumab but not everolimus.

Conclusion

TNFRSF9⁺ CD8⁺ T cells, which possessed both exhaustion and effector phenotype, were identified as an adverse prognosticator in ccRCC. These cells enrichment was associated with better immunotherapy response which indicated these cells potentially be crucial in immunotherapy.

Keywords:

• Tumor necrosis receptor super family 9
• Renal cell carcinoma
• Prognosis
• Immunotherapy
• CD8⁺ T cells

Acknowledgments

We thank Dr. Lingli Chen (Department of Pathology, Zhongshan Hospital, Fudan University, Shanghai, China) and Dr. Yunyi Kong (Department of Pathology, Fudan University Shanghai Cancer Center, Shanghai, China) for their excellent pathological technology help. The results are in part based upon data generated by the TCGA Research Network: https://www.cancer.gov/tcga.

Ethics approval and consent to participate

This study was approved by the Clinical Research Ethics Committee of Zhongshan Hospital, Fudan University, with the approval number B2015-030. Written informed consents were obtained from patients included.

Consent for publication:

All authors provide their consent for publication.

Data availability

All data generated that are relevant to the results presented in this article are included

in this article. Other data that were not relevant for the results presented here are

available from the corresponding author Dr. Xu upon reasonable request and ethically possible.

Conflict of interest

The authors have declared no conflicts of interest.

List of Abbreviations

ccRCC	=	clear cell renal carcinoma
TNFRSF9	=	Tumor necrosis factor receptor super family 9
ICB	=	immune check point blockade
TME	=	tumor microenvironment
OS	=	overall survival
RFS	=	recurrence free survival
HR	=	hazard ratio
CI	=	confidence interval

Supplemental material

Supplemental data for this article can be accessed on the publisher’s website.

Additional information

Funding

This work was supported by grants from National Natural Science Foundation of China [grant numbers: 31770851, 81702496, 81702497, 81702805, 81772696, 81872082, 81902556, 81902563, 81902898, 81974393], Shanghai Municipal Natural Science Foundation [grant numbers: 17ZR1405100, 19ZR1431800], Shanghai Sailing Program [grant numbers: 18YF1404500, 19YF1407900, 19YF1427200, 20YF1406100, 20YF1406200], Shanghai Municipal Commission of Health and Family Planning Program [grant numbers: 20174Y0042, 201840168, 20184Y0151] and Fudan University Shanghai Cancer Center for Outstanding Youth Scholars Foundation [grant number: YJYQ201802]. All these study sponsors have no roles in the study design, in the collection, analysis and interpretation of data.