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Updated: May 25, 2025
Pooled CRISPR-Based Genetic Screens in Mammalian Cells
Published on: September 4, 2019
Individualized Pooled CRISPR/Cas9 Screenings Identify CDK2 as a Druggable Vulnerability in a Canine Mammary Carcinoma
Marine Inglebert1,2, Martina Dettwiler1,3, Chang He1,2
1Institute of Animal Pathology, Vetsuisse Faculty, University of Bern, 3012 Bern, Switzerland.
Abstract:
High-throughput omics approaches have long been used to uncover potential vulnerabilities in human personalized oncology but are often limited by the lack of functional validation. Therefore, we placed our emphasis on functional drug testing using patient-derived organoids (PDOs). However, PDOs generated from tumors mostly lack comparison with matching normal tissue, and the number of testable drugs is limited. Here, we demonstrate how matching the neoplastic and non-neoplastic mammary PDOs derived from the same dog can utilize targeted CRISPR/Cas9 screens to unveil cancer cell specific vulnerabilities. We performed two independent CRISPR/Cas9 dropout screens using sub-libraries targeting the epigenome (n = 1269) or druggable genes (n = 834) in paired PDOs derived from both carcinoma and normal mammary tissues from the same dog. A comparison of essential genes for tumor cells survival identified CDK2 as a functional vulnerability in canine mammary tumors (CMTs) that can be targeted with the PF3600 inhibitor. Additional potential targets were also uncovered, providing insights for personalized cancer treatments in dogs.
Insights
This study introduces a novel method using paired canine organoids to identify cancer-specific vulnerabilities. Researchers found that targeting CDK2 with PF3600 may be a promising strategy for canine mammary tumors.
Area of Science:
- Comparative oncology
- Cancer genomics
- Drug discovery
Background:
- High-throughput omics lack functional validation in personalized oncology.
- Patient-derived organoids (PDOs) offer functional drug testing but often lack matched normal tissue.
- Limited drug screening capacity in current PDO models.
Purpose of the Study:
- To develop a method for functional validation of cancer vulnerabilities using paired neoplastic and non-neoplastic mammary PDOs from the same dog.
- To identify cancer-specific vulnerabilities in canine mammary tumors (CMTs) through targeted CRISPR/Cas9 screens.
- To explore potential therapeutic targets for personalized cancer treatment in dogs.
Main Methods:
- Generation of paired neoplastic and non-neoplastic mammary PDOs from canine patients.
- Two independent CRISPR/Cas9 dropout screens targeting the epigenome (1269 genes) and druggable genes (834 genes).
- Comparative analysis of essential genes for tumor cell survival versus normal mammary cells.
Main Results:
- Identification of CDK2 as a key vulnerability essential for CMT cell survival.
- Demonstration that PF3600, a CDK2 inhibitor, can target CMTs.
- Uncovering of additional potential therapeutic targets for CMTs.
Conclusions:
- Paired canine mammary PDOs combined with CRISPR/Cas9 screening provide a powerful platform for functional validation of cancer vulnerabilities.
- CDK2 is a validated therapeutic target for canine mammary tumors.
- This approach offers insights for developing personalized cancer therapies in veterinary medicine.
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