Determinants of anti-PD1 response and resistance in clear cell renal cell carcinoma

Abstract

Antigen recognition and T-cell mediated cytotoxicity in clear-cell renal cell carcinoma (ccRCC) remains incompletely understood. To address this knowledge gap, we analysed 115 multiregion tumour samples collected from 15 treatment-naive patients pre- and post-nivolumab therapy, and at autopsy in three patients. We performed whole-exome sequencing, RNAseq, TCRseq, multiplex immunofluorescence and flow cytometry analyses and correlated with clinical response. We observed pre-treatment intratumoural TCR clonal expansions suggesting pre-existing immunity. Nivolumab maintained pre-treatment expanded, clustered TCR clones in responders, suggesting ongoing antigen-driven stimulation of T-cells. T-cells in responders were enriched for expanded TCF7+CD8+ T-cells and upregulated GZMK/B upon nivolumab-binding. By contrast, nivolumab promoted accumulation of new TCR clones in non-responders, replacing pre-treatment expanded clonotypes. In this dataset, mutational features did not correlate with response to nivolumab and human endogenous retrovirus expression correlated indirectly. Our data suggests that nivolumab potentiates clinical responses in ccRCC by binding pre-existing expanded CD8+ T-cells to enhance cytotoxicity.

Histologic-Based Tumor-Associated Immune Cells Status in Clear Cell Renal Cell Carcinoma Correlates with Gene Signatures Related to Cancer Immunity and Clinical Outcomes

“…Since multiregional molecular genetic studies have largely been performed without consideration of morphology, the investigation regarding the correlation between histologic immune status and underlying genomes has been limited [8]. Genetic analysis and multiplex immunofluorescence/immunohistochemical staining remain challenging in routine clinical practice.…”

TCR sequencing: applications in immuno-oncology research

“…We have recently investigated this using paired longitudinal tumour and blood samples from patients with metastatic clear-cell renal cell carcinoma in a phase II study of first-line anti-PD-1 (nivolumab). 56 In bulk tumour samples, we showed that responders to nivolumab had significantly higher pre-treatment intratumour TCR clonality and cluster structure than non-responders, suggesting pre-existing adaptive immunity. Combining bulk and single-cell TCR and RNA-Seq data, we observed both novel and maintenance of pre-existing TCR clones in post-treatment samplesdbut only the latter correlated with ICB response.…”

“…Combining bulk and single-cell TCR and RNA-Seq data, we observed both novel and maintenance of pre-existing TCR clones in post-treatment samplesdbut only the latter correlated with ICB response. 56 Moreover, following treatment administration, we observed anti-PD-1 binds to both novel and pre-existing CD8þ T cells, but only pre-existing expanded TCR clones in responders had a cytotoxic phenotype (i.e. up-regulation of GZMB/K) and not in novel clones or in non-responder patients.…”

What We Have Learnt from CARMENA and SURTIME and What Should Be Done Differently in Future Trials on Cytoreductive Nephrectomy

“…The authors suggested that nivolumab drives both maintenance and replacement of previously expanded T cell clones. Only maintenance correlated with response [22].…”

“…Translational data from other tumour entities such as melanoma support using immunotherapy with the tumour antigens in place to prime the immune response, [20,21] but it is unclear if this is also required in the setting of metastatic RCC. A recent publication including patients with synchronous clear-cell mRCC who were pre-treated with nivolumab and underwent either a biopsy or CN suggests that nivolumab maintains a pre-existing Tcell mediated immune response in the tumour tissue of patients responding to therapy [22]. A higher level of T cells was found in responders both pre-and posttreatment.…”