Atg7 in CD4⁺ T cells improves intestinal mucosal inflammation by regulating Ets1-mediated T cell differentiation

Abstract

Background: Atg7-autophagy–related gene 7 contributes as an immune cell function regulator, particularly involved in CD4⁺ T cell response. Nevertheless, the specific contribution of Atg7 in CD4⁺ T cells sensitive immune responses in inflammatory bowel disease (IBD) remains largely unclear. This study explores the functional significance and regulatory mechanisms of CD4⁺ T cell-specific Atg7 in IBD.

Methods: Real-Time Quantitative Reverse Transcription PCR (qRT-PCR), western blotting analysis, flow cytometry and immunohistochemistry were employed to evaluate ATG7 expression in peripheral blood and colonic mucosal biopsies from IBD patients. Peripheral CD4⁺ T cells were transfected with lentivirus vectors encoding either ATG7 (LV-ATG7) or ATG7 short hairpin RNA (LV-shATG7). Furthermore, mice with a CD4⁺ T cell conditional knockout of Atg7 (Atg7^ΔCD4) were generated, and CD4⁺ T cells of splenic origin were subjected to RNA-seq.

Results: ATG7 level was markedly elevated in inflamed mucosa tissues and peripheral blood CD4⁺ T cells of patients with active Crohn's disease compared to healthy individuals. Overexpression of ATG7 suppressed Th1 differentiation while enhancing the induction of iTreg cells. Atg7^ΔCD4 mice exhibited exacerbated result of 2,4,6-trinitrobenzenesulphonic acid-induced experimental colitis, as did Rag1^−/− mice adoptively transferred with CD45^RBhigh CD4⁺ T cells from Atg7^ΔCD4 donors. RNA-seq analysis revealed that E26 transformation-specific sequence-1 (Ets1) acts downstream of Atg7 and mediates its regulatory effects on Th1 and Treg cell differentiation.

Conclusion: Our data indicate that Atg7 alleviates mucosal inflammation by modulating Th1/Treg differentiation through the regulation of Ets1 expression. Thus, CD4⁺ T cell-expressed Atg7 may serve as a promising therapeutic approach for IBD.

Keywords

Atg7 / autophagy / CD4⁺ T cells / IBD / Th1 differentiation

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