VHL loss reprograms the immune landscape to promote an inflammatory myeloid microenvironment in renal tumorigenesis
Melissa M. Wolf, … , W. Kimryn Rathmell, Jeffrey C. Rathmell
Melissa M. Wolf, … , W. Kimryn Rathmell, Jeffrey C. Rathmell
Published April 15, 2024
Citation Information: J Clin Invest. 2024;134(8):e173934. https://doi.org/10.1172/JCI173934.
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Research Article Metabolism Oncology

VHL loss reprograms the immune landscape to promote an inflammatory myeloid microenvironment in renal tumorigenesis

Abstract

Clear cell renal cell carcinoma (ccRCC) is characterized by dysregulated hypoxia signaling and a tumor microenvironment (TME) highly enriched in myeloid and lymphoid cells. Loss of the von Hippel Lindau (VHL) gene is a critical early event in ccRCC pathogenesis and promotes stabilization of HIF. Whether VHL loss in cancer cells affects immune cells in the TME remains unclear. Using Vhl WT and Vhl-KO in vivo murine kidney cancer Renca models, we found that Vhl-KO tumors were more infiltrated by immune cells. Tumor-associated macrophages (TAMs) from Vhl-deficient tumors demonstrated enhanced in vivo glucose consumption, phagocytosis, and inflammatory transcriptional signatures, whereas lymphocytes from Vhl-KO tumors showed reduced activation and a lower response to anti–programmed cell death 1 (anti–PD-1) therapy in vivo. The chemokine CX3CL1 was highly expressed in human ccRCC tumors and was associated with Vhl deficiency. Deletion of Cx3cl1 in cancer cells decreased myeloid cell infiltration associated with Vhl loss to provide a mechanism by which Vhl loss may have contributed to the altered immune landscape. Here, we identify cancer cell–specific genetic features that drove environmental reprogramming and shaped the tumor immune landscape, with therapeutic implications for the treatment of ccRCC.

Authors

Melissa M. Wolf, Matthew Z. Madden, Emily N. Arner, Jackie E. Bader, Xiang Ye, Logan Vlach, Megan L. Tigue, Madelyn D. Landis, Patrick B. Jonker, Zaid Hatem, KayLee K. Steiner, Dakim K. Gaines, Bradley I. Reinfeld, Emma S. Hathaway, Fuxue Xin, M. Noor Tantawy, Scott M. Haake, Eric Jonasch, Alexander Muir, Vivian L. Weiss, Kathryn E. Beckermann, W. Kimryn Rathmell, Jeffrey C. Rathmell

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Figure 4

Increased TAM displaying proinflammatory properties reside in the Vhl-KO TME.

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Increased TAM displaying proinflammatory properties reside in the Vhl-KO...
(A) Representative flow plots of myeloid cell populations in viable CD45+CD11b+ cell populations defined as PMN-MDSC (Ly6G+Ly6Clo-int), M-MDSC (Ly6G–Ly6Chi), overall TAMs (Ly6G–Ly6Clo), and the TAM subsets TAM1 (Ly6G–Ly6CloF4/80lo-int) and TAM2 (Ly6G–Ly6CloF4/80hi), and quantification of overall TAM, TAM1, and TAM2 infiltration as a percentage of viable cells in each Vhl WT and Vhl-KO pairs (pairs are represented by a matched symbol). See Supplemental Figure 3 for complete gating schemes. (B) Percentage of Phrodo+ cell populations as a fraction of viable CD45+CD11b+F4/80+ cells in Vhl WT.2 and Vhl-KO.7 tumors. Protein MFI quantification and representative histogram of (C) CD11c and (D) CD206 in overall TAMs from Vhl WT.2, Vhl-KO.7, Vhl rescue, and Vhl-KO.7 control tumors. (E) UMAP of CD45+ scRNA-Seq and mRNA expression levels of (F) Itgax (CD11c) and (G) Mrc1 (CD206) in TAMs/monocytes from Vhl WT.2 or KO.7 tumors. Each data point represents a biological replicate, and graphs show the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by ordinary 1-way ANOVA with Bonferroni’s multiple-comparison test (A), 2-tailed Student’s t test (B, C, and D), and Wilcoxon rank-sum test with a threshold of q < 0.05 (F and G). Max, maximum.