A Multispecies, Modality-Agnostic Scalable In Vivo Mosaic Screening Platform for Therapeutic Target Discovery.

Area of Science:

• Genomics
• Biotechnology
• Translational Medicine

Background:

• Validating therapeutic targets for complex diseases necessitates studying gene function within native tissue environments.
• In vitro models offer limited insights into in vivo disease pathology.

Purpose of the Study:

• To develop and validate a versatile in vivo high-throughput screening platform for identifying therapeutic targets in complex diseases.
• To establish a computational framework for analyzing functional genomics data against human disease signatures.

Main Methods:

• A modality-agnostic adeno-associated virus (AAV)-based platform was developed for in vivo delivery of genetic perturbations (knockouts, gain-of-function, synthetic miRNA knockdowns).
• High-dimensional screening data was analyzed using a curated framework that scores single-cell transcriptomes against human disease-specific molecular signatures.
• The platform was applied to a murine pulmonary fibrosis model and equine osteoarthritis joints.

Main Results:

• The screening platform successfully identified metabolic, antifibrotic, and immunomodulatory targets in both pulmonary fibrosis and osteoarthritis models.
• The analysis framework quantitatively ranked targets across diverse biological domains, including fibrosis and inflammation.
• Functional outcomes predicted by the framework were validated in orthogonal human ex vivo tissue models.

Conclusions:

• The developed in vivo screening platform and analysis framework provide a powerful paradigm for prioritizing therapeutic targets in complex diseases.
• This approach integrates high-dimensional in vivo functional genomics with human disease signatures for effective target discovery.
• The findings demonstrate the potential for accelerating the development of novel therapeutics by studying gene function in native diseased tissues.