A tumor-intrinsic PD-L1/NLRP3 inflammasome signaling pathway drives resistance to anti–PD-1 immunotherapy
Balamayoora Theivanthiran, … , Alisha Holtzhausen, Brent A. Hanks
Balamayoora Theivanthiran, … , Alisha Holtzhausen, Brent A. Hanks
Published February 4, 2020
Citation Information: J Clin Invest. 2020;130(5):2570-2586. https://doi.org/10.1172/JCI133055.
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Research Article Immunology Oncology

A tumor-intrinsic PD-L1/NLRP3 inflammasome signaling pathway drives resistance to anti–PD-1 immunotherapy

Abstract

An in-depth understanding of immune escape mechanisms in cancer is likely to lead to innovative advances in immunotherapeutic strategies. However, much remains unknown regarding these mechanisms and how they impact immunotherapy resistance. Using several preclinical tumor models as well as clinical specimens, we identified a mechanism whereby CD8+ T cell activation in response to programmed cell death 1 (PD-1) blockade induced a programmed death ligand 1/NOD-, LRR-, and pyrin domain–containing protein 3 (PD-L1/NLRP3) inflammasome signaling cascade that ultimately led to the recruitment of granulocytic myeloid-derived suppressor cells (PMN-MDSCs) into tumor tissues, thereby dampening the resulting antitumor immune response. The genetic and pharmacologic inhibition of NLRP3 suppressed PMN-MDSC tumor infiltration and significantly augmented the efficacy of anti–PD-1 antibody immunotherapy. This pathway therefore represents a tumor-intrinsic mechanism of adaptive resistance to anti–PD-1 checkpoint inhibitor immunotherapy and is a promising target for future translational research.

Authors

Balamayoora Theivanthiran, Kathy S. Evans, Nicholas C. DeVito, Michael Plebanek, Michael Sturdivant, Luke P. Wachsmuth, April K.S. Salama, Yubin Kang, David Hsu, Justin M. Balko, Douglas B. Johnson, Mark Starr, Andrew B. Nixon, Alisha Holtzhausen, Brent A. Hanks

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Figure 2

Wnt5a induces CXCR2-dependent chemokine expression in response to anti–PD-1 Ab immunotherapy.

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Wnt5a induces CXCR2-dependent chemokine expression in response to anti–P...
(A) TCGA human melanoma database gene expression analysis of CXCL5, CXCL2, and CXCR2 association with WNT5A. (B) Whole tumor tissue Western blot analysis of Wnt5a, YAP1, CXCL5, and vinculin and β-actin (used as loading controls). Blot is representative of 3 independent experiments. (C) Plasma CXCL5 ELISA following anti–PD-1 Ab therapy versus IgG isotype control therapy in the transgenic BRAFV600E PTEN–/– melanoma model (n = 6). Data are representative of 3 independent experiments. (D) qRT-PCR analysis of Cxcl1, Cxcl2, and Cxcl5 in the BRAFV600E PTEN–/– melanoma cell line following treatment with rWnt5a versus vehicle control (n = 3). (E) Western blot analysis of YAP1 expression in total cellular lysates (top) and nuclear lysates (middle) following treatment of BRAFV600E PTEN–/– melanoma cells with rWnt5a at various time points. Bottom blot shows Wnt5a induction of CXCL5 with or without verteporfin (YAP inhibitor) or XAV939 (β-catenin inhibitor). Blots shown are representative of 3 independent experiments. UT, untreated or vehicle control. (F) qRT-PCR analysis of Cxcl5 in BRAFV600E PTEN–/– NTC and Wnt5a-silenced BRAFV600E PTEN–/– melanoma cells (BRAFV600E PTEN–/– Wnt5aKD). Blot shows secreted CXCL5 in BRAFV600E PTEN–/– NTC and BRAFV600E PTEN–/– Wnt5aKD cells (n = 3). (G) IHC for CXCL5 (red) in BRAFV600E PTEN–/– NTC and BRAFV600E PTEN–/– Wnt5aKD tumor cells. Images are representative of 3 tumors. White arrows indicate CXCL5+ tumor cells. Original magnification, ×20. (H) IHC for Gr-1 in BRAFV600E PTEN–/– NTC and BRAFV600E PTEN–/– Wnt5aKD tumor cells. Original magnification, ×20. Plots show PMN-MDSC flow cytometric analysis of BRAFV600E PTEN–/– NTC and BRAFV600E PTEN–/– Wnt5aKD tumors (n = 3). (I) PMN-MDSC flow cytometric analysis of BRAFV600E PTEN–/– NTC and BRAFV600E PTEN–/– Wnt5aKD tumors following treatment with anti–PD-1 Ab versus IgG isotype control (n = 5). (J) Tumor volume change based on anti–PD-1 Ab/IgG control ratios for BRAFV600E PTEN–/– NTC and BRAFV600E PTEN–/– Wnt5aKD tumors (n = 5). α, anti. UT, untreated control. Kendall’s tau correlation coefficient was calculated for A. *P < 0.05 and ***P < 0.0005, by Student’s t test (C, D, and I) and 1-way ANOVA with Sidak’s post hoc multiple comparisons test (F). See also Supplemental Figure 3.