Autotaxin suppresses cytotoxic T cells via LPAR5 to promote anti–PD-1 resistance in non–small cell lung cancer
Jessica M. Konen, … , Jianjun Zhang, Don L. Gibbons
Jessica M. Konen, … , Jianjun Zhang, Don L. Gibbons
Published September 1, 2023
Citation Information: J Clin Invest. 2023;133(17):e163128. https://doi.org/10.1172/JCI163128.
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Research Article Immunology Oncology

Autotaxin suppresses cytotoxic T cells via LPAR5 to promote anti–PD-1 resistance in non–small cell lung cancer

Abstract

Non–small cell lung cancers that harbor concurrent KRAS and TP53 (KP) mutations are immunologically warm tumors with partial responsiveness to anti–PD-(L)1 blockade; however, most patients observe little or no durable clinical benefit. To identify novel tumor-driven resistance mechanisms, we developed a panel of KP murine lung cancer models with intrinsic resistance to anti–PD-1 and queried differential gene expression between these tumors and anti–PD-1–sensitive tumors. We found that the enzyme autotaxin (ATX), and the metabolite it produces, lysophosphatidic acid (LPA), were significantly upregulated in resistant tumors and that ATX directly modulated antitumor immunity, with its expression negatively correlating with total and effector tumor-infiltrating CD8+ T cells. Pharmacological inhibition of ATX, or the downstream receptor LPAR5, in combination with anti–PD-1 was sufficient to restore the antitumor immune response and efficaciously control lung tumor growth in multiple KP tumor models. Additionally, ATX was significantly correlated with inflammatory gene signatures, including a CD8+ cytolytic score in multiple lung adenocarcinoma patient data sets, suggesting that an activated tumor-immune microenvironment upregulates ATX and thus provides an opportunity for cotargeting to prevent acquired resistance to anti–PD-1 treatment. These data reveal the ATX/LPA axis as an immunosuppressive pathway that diminishes the immune checkpoint blockade response in lung cancer.

Authors

Jessica M. Konen, B. Leticia Rodriguez, Haoyi Wu, Jared J. Fradette, Laura Gibson, Lixia Diao, Jing Wang, Stephanie Schmidt, Ignacio I. Wistuba, Jianjun Zhang, Don L. Gibbons

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Figure 4

ATX expression negatively correlates with CD8+ T cell infiltration and effector status in tumors.

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ATX expression negatively correlates with CD8+ T cell infiltration and e...
(A) 344SQ-control (ctrl) or ATX-overexpressing cells were analyzed via Western blotting of cells and conditioned media (CM). ATX densitometric values were normalized to the corresponding actin or Ponceau bands and then to 344SQ-ctrl. (B) Tumor growth was measured from mice implanted with 344SQ-ctrl or -ATX cells and treated with IgG or anti–PD-1. n = 5 mice per group. *P < 0.05 and **P < 0.01, by multiple t tests (per time point). (C) Representative ATX and CD8 IHC images completed on tumors from B. CD8+ T cells were quantified as number per FOV. n = 3 mice each. *P < 0.05, **P < 0.01, and ****P < 0.0001, by 1-way ANOVA. Scale bars: 50 μm; insets zoomed 300% (ATX) or 250% (CD8). (D) 344SQ-ctrl or -ATX cells were cocultured with naive immune cells over time, and immune cell populations were analyzed by flow cytometry. The experiment was completed twice. *P < 0.05 and **P < 0.01, by t test. (E) 344SQPD1R2 cells depleted of ATX using 2 shRNAs or a control (scr) were analyzed as in A. (F) Tumor growth from 344SQPD1R2-scr and shATX#4 cells implanted into mice was monitored via calipers (left). Metastatic lung nodules were quantified at necropsy (right). n = 4–5 mice per group. *P < 0.05 and ***P < 0.001, by multiple t tests (G) Representative ATX and CD8 IHC images completed on tumors from F. n = 2 mice each, 6–9 FOV per tumor. ****P < 0.0001, by t test. Scale bars: 100 μm (ATX), 50 μm (CD8); insets zoomed 200%. (H) The 344SQPD1R2-scr and shATX cells from E were cocultured with naive immune cells as in D. The experiment was completed twice. *P < 0.05, **P < 0.01, and ***P < 0.001, by 1-way ANOVA.