Targeting T cell plasticity in kidney and gut inflammation by pooled single-cell CRISPR screening.

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_H17) 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_H17 cells could be ranked according to the resulting transcriptional perturbations, and polarization biases into T helper 1 (T_H1) 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_H17 to T_H1 cell plasticity in the human kidney in the context of renal autoimmunity. Editor's summary: Modulating plasticity in pathogenic T cells to promote a tolerogenic phenotype is a promising therapeutic approach in autoimmunity and cancer. To explore how pathogenic T_H17 cell plasticity is regulated in mouse models of kidney and gut inflammation, Enk et al. optimized an in vivo pooled single-cell CRISPR droplet sequencing (iCROP-seq) screen. iCROP-seq enabled the assessment and quantification of CRISPR-modified T_H17 cell polarization states in vivo. Furthermore, T cells from patients with antineutrophil cytoplasmic antibody (ANCA)–associated glomerulonephritis exhibited plasticity. These findings indicate that iCROP-seq can be applied to preclinical mouse models of inflammation and suggest that targeting T_H17 plasticity in patients with ANCA-glomerulonephritis is a promising therapeutic approach. —Hannah Isles [ABSTRACT FROM AUTHOR]