Pro-inflammatory CD4 + T cells are major drivers of autoimmune diseases, yet therapies modulating T cell phenotypes to promote an anti-inflammatory state are lacking. Here, we identify T helper 17 (T H 17) cell plasticity in the kidneys of patients with antineutrophil cytoplasmic antibody–associated glomerulonephritis on the basis of single-cell (sc) T cell receptor analysis and scRNA velocity. To uncover molecules driving T cell polarization and plasticity, we established an in vivo pooled scCRISPR droplet sequencing (iCROP-seq) screen and applied it to mouse models of glomerulonephritis and colitis. CRISPR-based gene targeting in T H 17 cells could be ranked according to the resulting transcriptional perturbations, and polarization biases into T helper 1 (T H 1) and regulatory T cells could be quantified. Furthermore, we show that iCROP-seq can facilitate the identification of therapeutic targets by efficient functional stratification of genes and pathways in a disease- and tissue-specific manner. These findings uncover T H 17 to T H 1 cell plasticity in the human kidney in the context of renal autoimmunity.

中文翻译：

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通过混合单细胞 CRISPR 筛选靶向肾脏和肠道炎症中的 T 细胞可塑性


促炎性 CD4 + T 细胞是自身免疫性疾病的主要驱动因素，但缺乏调节 T 细胞表型以促进抗炎状态的疗法。在这里，我们基于单细胞 (sc) T 细胞受体分析和 scRNA 速度，确定了抗中性粒细胞胞浆抗体相关性肾小球肾炎患者肾脏中的 T 辅助细胞 17 (TH 17) 细胞可塑性。为了揭示驱动 T 细胞极化和可塑性的分子，我们建立了体内混合 scCRISPR 液滴测序 (iCROP-seq) 筛选，并将其应用于肾小球肾炎和结肠炎的小鼠模型。 T H 17 细胞中基于 CRISPR 的基因靶向可以根据产生的转录扰动进行排序，并且可以量化 T 辅助细胞 1 (TH 1) 和调节性 T 细胞的极化偏差。此外，我们表明 iCROP-seq 可以通过以疾病和组织特异性的方式对基因和通路进行有效的功能分层，从而促进治疗靶点的识别。这些发现揭示了肾脏自身免疫背景下人肾中 T H 17 至 T H 1 细胞的可塑性。