In vivo identification of drug therapeutics for nemaline myopathy
Embargo Date
2028-02-11
OA Version
Citation
Abstract
This study employs the DiscoveryProbeTM FDA-approved drug library to perform a small molecule high-throughput drug screen as a strategy for developing therapies for Nemaline Myopathy (NM). Nemaline myopathy is a rare congenital neuromuscular disorder characterized by muscle weakness and the accumulation of fine rod-like structures, known as nemaline bodies, within skeletal muscle fibers. The main objective of this study was to identify potential therapeutic compounds capable of alleviating the pathological features of NM using a klhl41b-/- zebrafish model, a well-established representation of human NM pathology. 1320 FDA-approved drugs were screened using a novel drug repurposing strategy on the klhl41b-/- zebrafish model, an ortholog sharing 80% similarity with KLHL41. Core methodologies included weekly drug screening cycles which involved breeding and genotyping zebrafish lines, conducting behavioral assays with automated activity monitor to assess locomotor function, statistical analysis of the data using GraphPad Prism, and secondary validation screens to confirm drug efficacy. The preliminary screen identified 56 compounds that significantly improved the swimming behavior in the klhl41b zebrafish model, 830 compounds that showed no significance, 161 compounds that resulted in a reduction in swimming behavior, and 273 that displayed toxicity. These 56 compounds improving swimming were further analyzed to uncover common molecular pathways, revealing key mechanisms of action including adrenergic receptors, cholinergic receptors, histamine receptors, G-protein coupled receptors/ion channels, nuclear receptors, tyrosine kinase receptors, and other non-receptor enzyme inhibitors. To test the efficacy of these compounds in a mammalian system, selected candidates will be further screened in Klhl41 mutant C2C12 myoblast cells, bridging the gap between zebrafish models and mammalian systems. This study highlights the effectiveness of a small molecule high-throughput drug screening approach in identifying promising therapeutics for NM. It lays the groundwork for advancing these candidates toward clinical applications.
Description
2025