JCI Insight - Impaired regulation by purinergic signaling axis contributes to CD8+ T cell dysregulation in STAT3 gain of function

Impaired regulation by purinergic signaling axis contributes to CD8⁺ T cell dysregulation in STAT3 gain of function

Jose S. Campos Duran, Montana S. Knight, Samir U. Sayed, Megan C. Dalalo, Andrea A. Mauracher, Peyton Conrey, Aaron B. Schultz, Ceire A. Hay, Robert B. Lindell, Ilona Neale, Kyle Yeakle, Eric D. Abrams, Erica G. Schmitt, Martin A. Thelin, Christian A. Howard, Sara Bluestein, Christine M. Seroogy, Tamara C. Pozos, Akaluck Thatayatikom, Ingrid S. Lundgren, Amelie Gauthier, Scott W. Canna, Helen C. Su, Michael D. Keller, Ottavia M. Delmonte, Lisa R. Forbes Satter, Steven M. Holland, Jenna R.E. Bergerson, Jennifer W. Leiding, Neil Romberg, Will Bailis, Christopher A. Hunter, Alexandra F. Freeman, Alejandro V. Villarino, Mark S. Anderson, Megan A. Cooper, Tiphanie P. Vogel, Sarah E. Henrickson

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Research Article Immunology Metabolism

Impaired regulation by purinergic signaling axis contributes to CD8⁺ T cell dysregulation in STAT3 gain of function

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Abstract

Gain-of-function (GOF) variants in STAT3 cause a complex disorder characterized by early-onset autoimmunity, lymphoproliferation, recurrent infections, and immune dysregulation. In both primary human and mouse models of STAT3 GOF, CD8+ T cells have been implicated as pathogenic drivers of autoimmunity, though the exact mechanisms remain poorly understood. Here, we found that in patients with STAT3 GOF, CD8+ T cells exist in an activated state. Functional assessment revealed that naive CD8+ T cells have an increased capacity for IFN-γ and TNF-α production, with type I and type II IFN transcriptional signatures. Evaluation of immunoregulatory pathways revealed dysregulation of the purinergic signaling axis in CD8+ T cells: CD39 was increased, whereas downstream purinergic family members, CD73 and the adenosine receptor A2AR, were downregulated, impairing the potential to produce or sense immunosuppressive adenosine. Evaluation of the impact of precision therapy, in the form of JAK inhibition, at a cellular and functional level revealed partial normalization of CD8+ T cell dysregulation in patients, including aberrant cytokine production. Our study suggests that a dysregulated purinergic signaling axis plays a key role in CD8+ T cell dysregulation in STAT3 GOF and may have implications for other rare monogenic immune disorders and common inflammatory disorders.

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Figure 3

Dysregulation of purinergic pathway in patients with STAT3 GOF.

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(A) Flow cytometry plot and quantification of CD39 frequency within CD8⁺ T cells in healthy controls (n = 47) and patients with STAT3 GOF (n = 14). (B) Schematic of purinergic signaling regulation of immune cell activation and function. (C) CD39 (ENTPD1), CD73 (NT5E), and A_2AR (ADORA2A) row z score and log₂ count per million (log₂ CPM) in CD8⁺ T cells from scRNA-Seq studies. (D) Representative flow cytometry plot and CD73 frequency on CD8⁺ T cells from healthy controls (n = 42) and untreated patients with STAT3 GOF (n = 10). (E) Representative histogram of A_2AR expression on CD8⁺ T cells in healthy controls (n = 18) and untreated patients with STAT3 GOF (n = 5). (F) Schematic of 14-day in vitro culture of healthy control CD8⁺ T cells activated with αCD3/αCD28 with or without IL-21 or IL-27. Time course of (G) CD39, (H) CD73, and (I) A_2AR induction over 14 days followed by frequency (n = 10) and MFI quantification (n = 4) at day 14. Data are pooled from more than 3 independent experiments for A, D, and G–I, and from 2 experiments for E. Data represent mean ± SEM. For A, D, and E: *P ≤ 0.05 and **P ≤ 0.01 by Mann-Whitney test. For G–I: *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001 by repeated-measures 1-way ANOVA with Tukey’s multiple-comparison test. Figure 3B (https://BioRender.com/otmaeli) and Figure 3F (https://BioRender.com/lmy3557) were created in BioRender.

Impaired regulation by purinergic signaling axis contributes to CD8+ T cell dysregulation in STAT3 gain of function

Impaired regulation by purinergic signaling axis contributes to CD8+ T cell dysregulation in STAT3 gain of function

Abstract

Authors

Figure 3

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Impaired regulation by purinergic signaling axis contributes to CD8⁺ T cell dysregulation in STAT3 gain of function

Impaired regulation by purinergic signaling axis contributes to CD8⁺ T cell dysregulation in STAT3 gain of function