The Intratumoral Balance between Metabolic and Immunologic Gene Expression Is Associated with Anti–PD-1 Response in Patients with Renal Cell Carcinoma

Abstract

Pretreatment tumor PD-L1 expression correlates with response to anti-PD-1/PD-L1 therapies. Yet, most patients with PD-L1+ tumors do not respond to treatment. The current study was undertaken to investigate mechanisms underlying the failure of PD-1–targeted therapies in patients with advanced renal cell carcinoma (RCC) whose tumors express PD-L1. Formalin-fixed, paraffin-embedded (FFPE) pretreatment tumor biopsies expressing PD-L1 were derived from 13 RCC patients. RNA was isolated from PD-L1+ regions and subjected to whole genome microarray and multiplex quantitative (q)RT-PCR gene expression analysis. A balance between gene expression profiles reflecting metabolic pathways and immune functions was associated with clinical outcomes following anti-PD-1 therapy. In particular, the expression of genes involved in metabolic and solute transport functions such as UGT1A family members, also found in kidney cancer cell lines, was associated with treatment failure in patients with PD-L1+ RCC. Conversely, tumors from responding patients overexpressed immune markers such as BACH2, a regulator of CD4+ T cell differentiation, and CCL3, involved in leukocyte migration. These findings suggest that tumor cell–intrinsic metabolic factors may contribute to treatment resistance in RCC, thus serving as predictive markers for treatment outcomes and potential new targets for combination therapy regimens with anti-PD-1.

Th17 immune microenvironment in Epstein-Barr virus–negative Hodgkin lymphoma: implications for immunotherapy

“…13,14 However, some CHLs are resistant to anti-PD-1 monotherapy, or they relapse after an initial response, raising the question of whether more effective anti-PD-1-containing treatment combinations could be rationally devised based on immunologic and gene expression analyses of the CHL TME. 22,37 For example, although the LAG3 gene was overexpressed in EBV 1 CHL, every tumor in our study expressed LAG-3 protein on $1% of cells by IHC (Figure 1), supporting current clinical testing of the combination of anti-PD-1 and anti-LAG-3 in CHL (NCT02061761). In addition, because the EBV residing in some CHLs is strongly immunogenic, we hypothesized that differences in the immune TME of EBV 1 vs EBV 2 CHL would have specific implications for designing combination treatment regimens.…”

“…Tissue was macrodissected by scalpel from 5-mm sections, as described. 22 RNA was isolated with the High Pure RNA Paraffin Kit (Roche, Indianapolis, IN), as described. 22,23 Seventyfive nanograms of total RNA was reverse transcribed in a 10-mL reaction volume using qScript cDNA SuperMix (Quanta Biosciences, Gaithersburg, MD) per protocol.…”

“…22 RNA was isolated with the High Pure RNA Paraffin Kit (Roche, Indianapolis, IN), as described. 22,23 Seventyfive nanograms of total RNA was reverse transcribed in a 10-mL reaction volume using qScript cDNA SuperMix (Quanta Biosciences, Gaithersburg, MD) per protocol. From each RT reaction, 7.5 mL was preamplified in a total volume of 30 mL using a 14-cycle PCR reaction per PreAmp protocol (Applied Biosystems, Foster City, CA).…”

Mitochondrial dysregulation and glycolytic insufficiency functionally impair CD8 T cells infiltrating human renal cell carcinoma

“…The VHL mutation permits stabilization of hypoxia inducible factors (HIF-1α and HIF-2α), transcription factors that promote tumor cell glycolysis, and altered metabolism through altered transcriptional programs. This altered metabolic environment may influence outcomes following PD-1 blockade therapy (30). Here, we assessed the functionality and cell intrinsic metabolism of ccRCC TIL from 54 patients.…”

Mesenchymal traits at the convergence of tumor-intrinsic and -extrinsic mechanisms of resistance to immune checkpoint blockers

“…ICB therapy restores glucose in the TME, permitting T-cell glycolysis and IFN-γ production [162]. Of clinical relevance, the up-regulation of genes corresponding to proteins associated with metabolic and solute transport functions has been found in tumors of renal cell carcinoma patients, nonresponding to anti-PD-1 therapy [163], confirming the involvement of an altered metabolism in response to ICB-based immunotherapy.…”