C. elegans as a model organism for human disease through the application of phenologs

National Center For Advancing Translational Sciences

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Abstract

This project has advanced from our Ipglycermides: Novel potent and selective inhibitors of parasitic phosphoglycerate mutase project where C. elegans serve as a model organism for infectious nematode species to test the activity of membrane permeable cyclic peptides on the viability of the organism. The ADST laboratory has developed a 384-well microtiter plate, laser cytometry-based qHTS assay platform validated using a chemical library of anti-infective agents to identify those with anti-nematodal activity. Primary screening is based on an overall decrease in worm number and area as measured by green fluorescence in a strain of worm ubiquitously expressing GFP. The lab is equipped with the necessary instrumentation for follow-up assays involving high-content imaging, life stage sorting, and collaborator-enabled proteomics analysis to distinguish various modes-of-action for the potential chemical modulators. In collaboration with Dr. D. Sibley (NINDS, NIH) the ADST laboratory has developed a 384-well qHTS laser cytometry-based screening assay for Parkinsons Disease evaluating neurodegeneration of dopaminergic neurons in C. elegans strains containing two different human PD-linked genes. Both strains express GFP in their dopaminergic neurons allowing for traceable fluorescence intensity as a measure of neuron health over time. Preliminary data has been reporter at the 2023 The American Society for Pharmacology and Experimental Therapeutics Annual meeting (abstract below by Ash CT et al.). In a collaboration with Dr. Mackenzies laboratory (U. Rochester Medical Center) the ADST lab will develop a C. elegans qHTS assay to investigate the genotypic basis for the TANGO2 deficiency disorder (TDD) phenotype. TDD is an autosomal recessive neurodegenerative condition. C. elegans harbors two homologs of the TANGO2 gene, hrg-9 and hrg-10. Worms with biallelic pathogenic variants in these genes exhibit a strong behavioral phenotype that involves reduced brood size and low motility. We will exploit these characteristics to determine whether certain pharmacological compounds reverse or ameliorate the disease phenotype. The TDD C. elegans human phenolog will be engineered to harbor one or more endogenously expressed fluorescent proteins to enable in vivo quantitative screening (qHTS) of drug and investigational agent chemical libraries. Lead compounds will be tested in future experiments with the hope that they may also be beneficial for patients with TDD. Dr. Mackenzies laboratory has generated a single knockout strain of hrg-10-/- mutants and acquired an hrg-9-/- knockout strain from the Caenorhabditis Genetics Center (CGC). His lab is in the process of generating a double knockout strain that also expresses a whole-body fluorescent reporter.