The Endo-GeneScreen platform identifies drug-like probes that regulate endogenous protein levels within physiological...

Area of Science:

• Biochemistry
• Genetics
• Pharmacology

Background:

• Traditional drug discovery has limitations in scalability and mechanistic insight.
• Endogenous protein level regulation is crucial for disease understanding and therapeutic intervention.

Purpose of the Study:

• To develop a high-throughput platform, Endo-GeneScreen (EGS), for identifying small molecules that modulate endogenous protein levels.
• To apply EGS to discover modulators of Syngap1, a gene implicated in neurodevelopmental disorders.

Main Methods:

• EGS platform development for high-throughput screening of small molecules.
• Application of EGS in disease-modeling contexts to identify compounds affecting Syngap1 protein levels.
• Validation of identified compounds and characterization of their mechanism of action.

Main Results:

• >40 validated small molecules identified that increase endogenous neuronal Syngap1 levels.
• SR-1815, a novel multikinase inhibitor, was validated as a proof-of-concept candidate.
• SR-1815 restores SynGAP protein to wildtype levels and mitigates loss-of-function consequences by regulating Syngap1 splicing.

Conclusions:

• The Endo-GeneScreen platform enables efficient identification and preclinical development of small molecules targeting endogenous protein abundance.
• EGS accelerates the discovery of therapeutic agents for diseases linked to protein level dysregulation, such as neurodevelopmental disorders.
• The platform facilitates mechanistic deconvolution of drug candidates and orphaned probes in relevant biological contexts.