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A Whole Genome-Wide Arrayed CRISPR Screen in Primary Organ Fibroblasts to Identify Regulators of Kidney Fibrosis
Robert J Turner1, Stefan Golz2, Carina Wollnik1
1In Vitro Biology, Evotec SE, Hamburg, Germany.
Abstract:
Kidney fibrosis presents a hallmark of chronic kidney disease. With ever-increasing patient numbers and limited treatment options available, novel strategies for therapeutic intervention in kidney disease are warranted. Fibrosis commonly results from a wound healing response to repeated or chronic tissue damage, irrespective of the underlying etiology, and can occur in virtually any solid organ or tissue. In order to identify targets relevant for kidney fibrosis, we aimed to employ CRISPR screening in primary human kidney fibroblasts. We demonstrate that CRISPR technology can be applied in primary kidney fibroblasts and can furthermore be used to conduct arrayed CRISPR screening using a high-content imaging readout in a whole genome-wide manner. Hits coming out of this screen were validated using orthogonal approaches and present starting points for validation of novel targets relevant to kidney disease.
Insights
Researchers used CRISPR screening in human kidney cells to find new ways to treat kidney fibrosis, a common problem in chronic kidney disease. This study identifies potential targets for developing novel therapies for kidney disorders.
Area of Science:
- Nephrology
- Molecular Biology
- Genomics
Background:
- Kidney fibrosis is a key feature of chronic kidney disease (CKD), affecting a growing patient population with limited therapeutic options.
- Fibrosis arises from wound healing responses to persistent kidney damage, regardless of the initial cause.
- Identifying novel therapeutic targets is crucial for managing kidney fibrosis and CKD progression.
Purpose of the Study:
- To apply CRISPR screening technology in primary human kidney fibroblasts to identify novel targets for kidney fibrosis.
- To establish and validate CRISPR screening methods for use in kidney fibroblast research.
Main Methods:
- Utilized CRISPR screening technology in primary human kidney fibroblasts.
- Performed arrayed, whole genome-wide CRISPR screening with a high-content imaging readout.
- Validated screening hits using orthogonal experimental approaches.
Main Results:
- Demonstrated the successful application of CRISPR technology in primary human kidney fibroblasts.
- Successfully conducted arrayed, whole genome-wide CRISPR screening in this cellular model.
- Identified validated hits from the screen, providing starting points for further target investigation.
Conclusions:
- CRISPR screening is a viable and powerful tool for dissecting molecular pathways in kidney fibrosis.
- The identified targets represent promising avenues for the development of future kidney disease therapeutics.
- This approach facilitates the discovery of novel strategies to combat kidney fibrosis.
