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

Sci Immunol. 2024 Jun 14;9(96):eadd6774. doi: 10.1126/sciimmunol.add6774. Epub 2024 Jun 14.

Abstract

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 (TH17) 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 TH17 cells could be ranked according to the resulting transcriptional perturbations, and polarization biases into T helper 1 (TH1) 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 TH17 to TH1 cell plasticity in the human kidney in the context of renal autoimmunity.

MeSH terms

  • Animals
  • CRISPR-Cas Systems
  • Cell Plasticity / genetics
  • Cell Plasticity / immunology
  • Clustered Regularly Interspaced Short Palindromic Repeats / immunology
  • Colitis / genetics
  • Colitis / immunology
  • Female
  • Glomerulonephritis / genetics
  • Glomerulonephritis / immunology
  • Humans
  • Inflammation / genetics
  • Inflammation / immunology
  • Kidney / immunology
  • Kidney / pathology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Single-Cell Analysis*
  • Th17 Cells* / immunology