Abstract: TH-PO558
Targeting Tissue-Specific T Cell Plasticity by Pooled Single-Cell CRISPR Screening in Preclinical Mouse Models
Session Information
- Glomerular Diseases: From Inflammation to Fibrosis - I
November 02, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1401 Glomerular Diseases: From Inflammation to Fibrosis
Authors
- Hellmig, Malte, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Riecken, Kristoffer, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Paust, Hans-Joachim, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Zhao, Yu, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Jauch-Speer, Saskia-L., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Sivayoganathan, Varshi, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Liu, Shuya, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Wiech, Thorsten, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Huber, Tobias B., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Bonn, Stefan, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Huber, Samuel, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Panzer, Ulf, Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
- Krebs, Christian F., Universitatsklinikum Hamburg-Eppendorf, Hamburg, Hamburg, Germany
Background
Treatment of autoimmune diseases demands a shift from unspecific immunosuppression towards targeted therapies. This could be achieved by turning pro-inflammatory T helper cells into anti-inflammatory subsets. However, the molecular pathways involved in T cell plasticity and stability are not fully understood. Single cell CRISPR-screens are a powerful tool to simultaneously analyze the impact of multiple genes on cellular phenotypes.
Methods
By combining single cell gene expression analysis and T cell receptor sequencing, we uncover Th17 to Th1 cell plasticity in the human kidney in renal autoimmunity. To investigate the molecules involved in T cell plasticity in disease settings, we established in vivo single cell CRISPR droplet sequencing (iCROP-seq).
Results
By applying this technique to in vivo models of inflammatory diseases in the kidney and intestine, we demonstrate that CRISPR-induced alterations in T cell polarization can be identified and ranked according to corresponding transcriptional perturbations. In particular, we targeted pro-inflammatory Th17 cells in models of immune-mediated diseases and quantified polarization biases into Th1 and regulatory T cells.
Conclusion
iCROP-seq will facilitate the identification of therapeutic targets by highly efficient functional stratification of genes and pathways in a disease- and tissue-specific manner.
Funding
- Government Support – Non-U.S.