FOXO1 pathway activation by VISTA immune checkpoint restrains pulmonary ILC2 functions
Mohammad Hossein Kazemi, … , Kei Sakano, Omid Akbari
Mohammad Hossein Kazemi, … , Kei Sakano, Omid Akbari
Published January 2, 2025
Citation Information: J Clin Invest. 2025;135(4):e184932. https://doi.org/10.1172/JCI184932.
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Research Article Immunology Inflammation

FOXO1 pathway activation by VISTA immune checkpoint restrains pulmonary ILC2 functions

Abstract

Group 2 innate lymphoid cells (ILC2s) play a pivotal role in the development of airway hyperreactivity (AHR). However, the regulatory mechanisms governing ILC2 function remain inadequately explored. This study uncovers V-domain Ig suppressor of T cell activation (VISTA) as an inhibitory immune checkpoint crucial for modulating ILC2-driven lung inflammation. VISTA is upregulated in activated pulmonary ILC2s and plays a key role in regulating lung inflammation, as VISTA-deficient ILC2s demonstrate increased proliferation and function, resulting in elevated type 2 cytokine production and exacerbation of AHR. Mechanistically, VISTA stimulation activates Forkhead box O1 (FOXO1), leading to modulation of ILC2 proliferation and function. The suppressive effects of FOXO1 on ILC2 effector function were confirmed using FOXO1 inhibitors and activators. Moreover, VISTA-deficient ILC2s exhibit enhanced fatty acid oxidation and oxidative phosphorylation to meet their high energy demands. Therapeutically, VISTA agonist treatment reduces ILC2 function both ex vivo and in vivo, significantly alleviating ILC2-driven AHR. Our murine findings were validated in human ILC2s, whose function was reduced ex vivo by a VISTA agonist, and in a humanized mouse model of ILC2-driven AHR. Our studies unravel VISTA as an immune checkpoint for ILC2 regulation via the FOXO1 pathway, presenting potential therapeutic strategies for allergic asthma by modulating ILC2 responses.

Authors

Mohammad Hossein Kazemi, Zahra Momeni-Varposhti, Xin Li, Benjamin P. Hurrell, Yoshihiro Sakano, Stephen Shen, Pedram Shafiei-Jahani, Kei Sakano, Omid Akbari

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Figure 6

ILC2s lacking VISTA exhibit higher mitochondrial respiration and FAO.

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ILC2s lacking VISTA exhibit higher mitochondrial respiration and FAO. (A...
(A and B) WT and VISTA-deficient mice received 3 days of intranasal rmIL-33. Total RNA was extracted from isolated pulmonary aILC2s to perform a bulk transcriptomic analysis. (A) Chord plot representing the most highly differentially expressed genes from top upregulated metabolic pathways. Specific pathways are color-coded and represented in the right inner bands, where chords gather. Outer bands (yellow to red) on the right depict the IPA –log10 P value. The left inner bands (white to red) represent the gene –log10 P value. The left outer bands (white to purple) represent the gene log2(fold change). (B) Schematic of TCA cycle and FAO showing the trends of enzymes differentially modulated in Vsir–/– versus WT aILC2s, with the color-coding trends of intermediate metabolites based on IPA analysis (blue to orange for inhibition to activation). (C–L) WT and Vsir–/– aILC2s underwent metabolic analyses. (D) Bar plots showing the quantification of MitoTracker Green (as MFI). (E) Bar plots showing the quantification of TMRM in freshly isolated ILC2s (as MFI). (F and G) BODIPY FL C16 (F) and BODIPY493/503 (G) expression levels (as MFI). (H–L) Mitochondrial respiratory profile. (H) Oxygen consumption rate (OCR) in response to oligomycin, BAM15, and rotenone plus antimycin A (Rot/AA) sequential injections. (I–L) Basal respiration (I), spare respiratory capacity (J), ATP production rate (K), and mitochondrial ATP (mito-ATP) and glycolytic ATP (glycol-ATP) production (L). Data are presented as mean + SEM and are representative of at least 2 independent experiments. Statistical significance was assessed using 2-tailed Student’s t test; *P < 0.05, **P < 0.01, ***P < 0.001.