Functional genetic dropout screens and in vivo validation of candidate therapeutic targets using mouse mammary tumoroids

Area of Science:

• Oncology
• Genetics
• Biotechnology

Background:

• BRCA1/2-deficient breast cancer poses a significant therapeutic challenge.
• Three-dimensional (3D) tumoroids derived from genetically engineered mouse models (GEMMs) offer a promising preclinical platform.
• Existing methods lack efficient genetic manipulation and in vivo validation strategies for these tumoroids.

Purpose of the Study:

• To present a comprehensive protocol for genetic modification of BRCA1/2-deficient breast cancer tumoroids.
• To enable functional genetic dropout screens using these modified tumoroids.
• To facilitate rapid and straightforward in vivo validation of potential therapeutic targets identified through screening.

Main Methods:

• Development of genetically engineered mouse models (GEMMs) for BRCA1/2-deficient breast cancer.
• Establishment and genetic modification of 3D tumoroids derived from GEMMs.
• Execution of functional genetic dropout screens within the modified tumoroids.
• Orthotopic transplantation of modified tumoroids for in vivo studies.

Main Results:

• The protocol successfully allows for genetic modification of 3D tumoroids.
• Functional genetic dropout screens can be effectively performed using the modified tumoroids.
• Orthotopic transplantation enables robust in vivo validation of candidate therapeutic targets.

Conclusions:

• This protocol provides a powerful tool for identifying and validating therapeutic targets in BRCA1/2-deficient breast cancer.
• The described methodology accelerates the drug discovery and development pipeline for this disease.
• The 3D tumoroid system combined with genetic screening and in vivo validation offers a significant advancement in preclinical cancer research.