FLECS technology for high-throughput screening of hypercontractile cellular phenotypes in fibrosis: A function-first approach to anti-fibrotic drug discovery.

The pivotal role of myofibroblast contractility in the pathophysiology of fibrosis is widely recognized, yet HTS approaches are not available to quantify this critically important function in drug discovery. We developed, validated, and scaled-up a HTS platform that quantifies contractile function of primary human lung myofibroblasts upon treatment with pro-fibrotic TGF-β1. With the fully automated assay we screened a library of 40,000 novel small molecules in under 80 h of total assay run-time. We identified 42 hit compounds that inhibited the TGF-β1-induced contractile phenotype of myofibroblasts, and enriched for 19 that specifically target myofibroblasts but not phenotypically related smooth muscle cells. Selected hits were validated in an ex vivo lung tissue models for their inhibitory effects on fibrotic gene upregulation by TGF-β1. Our results demonstrate that integrating a functional contraction test into the drug screening process is key to identify compounds with targeted and diverse activity as potential anti-fibrotic agents.
Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ivan Pushkarsky reports a relationship with Forcyte Biotechnologies, Inc that includes: board membership, employment, and equity or stocks. Robert Damoiseaux reports a relationship with Forcyte Biotechnologies, Inc that includes: consulting or advisory and equity or stocks. Boris Hinz reports a relationship with Forcyte Biotechnologies, Inc that includes: consulting or advisory and equity or stocks. Yao Wang reports a relationship with Forcyte Biotechnologies, Inc that includes: employment and equity or stocks. Enrico Cortes reports a relationship with Forcyte Biotechnologies, Inc that includes: employment and equity or stocks. Ricky Huang reports a relationship with Forcyte Biotechnologies, Inc that includes: employment and equity or stocks. Jeremy Wan reports a relationship with Forcyte Biotechnologies, Inc that includes: employment and equity or stocks. Ivan Pushkarsky has patent #US11592438B2 issued to UCLA. Robert Damoiseaux is an employee at UCLA which holds the patent licensed by Forcyte which pertains to research presented in this work.
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