UBA1-depleted neutrophils disrupt immune homeostasis and induce VEXAS-like autoinflammatory disease in mice
Ge Dong, Jingjing Liu, Wenyan Jin, Hongxi Zhou, Yuchen Wen, Zhiqin Wang, Keyao Xia, Jianlin Zhang, Linxiang Ma, Yunxi Ma, Lorie Chen Cai, Qiufan Zhou, Huaquan Wang, Wei Wei, Ying Fu, Zhigang Cai
Ge Dong, Jingjing Liu, Wenyan Jin, Hongxi Zhou, Yuchen Wen, Zhiqin Wang, Keyao Xia, Jianlin Zhang, Linxiang Ma, Yunxi Ma, Lorie Chen Cai, Qiufan Zhou, Huaquan Wang, Wei Wei, Ying Fu, Zhigang Cai
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Research Article Hematology Inflammation

UBA1-depleted neutrophils disrupt immune homeostasis and induce VEXAS-like autoinflammatory disease in mice

Abstract

Vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic (VEXAS) syndrome is a hemato-rheumatoid disease caused by somatic UBA1 mutations in hematopoietic stem cells (HSCs). The pathogenic cell type(s) responsible for the syndrome are unknown, and murine models recapitulating the disease are lacking. We report that loss of Uba1 in various mouse hematopoietic cell types resulted in pleiotropic consequences and demonstrate that an approximate 70% loss of Uba1 in neutrophils (NEs) of murine mutants induced nonlethal VEXAS-like symptoms. Depletion of Uba1 in HSCs induced extensive hematopoietic cell loss, whereas depletion of Uba1 in B cells, T cells, or megakaryocytes induced corresponding cell death, but these mutant mice appeared normal. Depletion of Uba1 in monocytes and NEs failed to induce cell death, and the mutant mice were viable. Among the tested models, only depletion of Uba1 in NEs induced autoinflammatory symptoms including increased counts and percentages of NEs, increased proinflammatory cytokines, presence of vacuoles in myeloid cells, splenomegaly, and dermatitis. Residual Uba1 was approximately 30% in the mutant NEs, which disrupted cellular hemostasis. Finally, genetic loss of the myeloid prosurvival regulator Morrbid partially mitigated the VEXAS-like symptoms. The established VEXAS-like murine model will further our understanding and treatment of the newly identified autoinflammatory syndrome prevalent among aged men.

Authors

Ge Dong, Jingjing Liu, Wenyan Jin, Hongxi Zhou, Yuchen Wen, Zhiqin Wang, Keyao Xia, Jianlin Zhang, Linxiang Ma, Yunxi Ma, Lorie Chen Cai, Qiufan Zhou, Huaquan Wang, Wei Wei, Ying Fu, Zhigang Cai

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Figure 7

Cell-autonomous proinflammatory activation in S100a8Cre-CKO NEs.

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Cell-autonomous proinflammatory activation in S100a8Cre-CKO NEs. (A) Mea...
(A) Measurement of ROS production in NEs by flow cytometry. NEs from BM were gated by Gr1+Cd11b+ staining before the ROS analysis. Left panel shows representative flow cytometric profiles. Right panel shows quantitative results for ROShi NEs and ROS expression levels. n = 4 biological repeats per group. MFI, mean florescence of intensity. (B) Western blot analysis of inflammatory markers. MPO is a classic readout of inflammation; IL-6 and TNF-α are proinflammatory cytokines; Nlrp3 and caspase-1 are important components of the inflammasome complex or pathways. The fold changes of the tested proteins are between the ranges of 1.5 (caspase-1) and 7.25 (TNF-α). P values were labeled for each marker on the panel. R1/R2/R3 indicates 3 independent biological repeats. (C) Secretion and intracellular levels of the proinflammatory cytokines measured by ELISA. To measure the secretion level, 1 × 106 purified NEs were cultured for 24 hours in a 96-well plate with 100 μL medium. To measure intracellular levels, 1 × 107 purified NEs were incubated with RIPA lysis buffer. n = 4~6 biological repeats per group. Data represent the mean ±SEM. *P < 0.05, **P < 0.01, and ****P < 0.0001, by Student’s t-test. n = 3~6 biological repeats.