Discovery of a Novel Probe for GPR-Sixty-Five and Its Inflammatory Bowel Disease Risk Variant [electronic resource].

9780355027952

Ann Arbor : ProQuest Dissertations & Theses, 2017

1 online resource (111 p.)

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Inflammatory Bowel Disease (IBD) is a chronic immune disorder affecting the intestines. Genome-wide association scans (GWAS) of diseased versus healthy tissues have identified 163 susceptibility loci, and have also revealed links between the gut microbiome, autophagy, and IBD. GPR65 is a proton-sensing G protein-coupled receptor (GPCR) identified by GWAS and functional genomic investigations as a protein important for proper lysosomal function, bacterial autophagy, and anti-inflammatory effects. GWAS identified an isoleucine-to-leucine variant of GPR65, I231L, which confers increased risk of developing IBD. GPR65 knock down and knock out models have provided a wealth of information about the mechanism of action of GPR65's connection to lysosomal and autophagic function; however, the ability to stimulate GPR65 for further study is currently limited. Current small molecule agonists of GPR65, such as BTB09089, lose efficacy at lower pHs, which limits the scope of their use in settings such as sites of inflammation where acidosis causes acidic microenvironments. This thesis describes the discovery of the first positive allosteric modulator of GPR65, BRD2813, through high-throughput screening and secondary assays. BRD2813 is also the first small molecule shown to stimulate the 1231L variant of GPR65. BRD2813 revealed a novel binding pocket in GPR65, and highlighted a possible difference in activity between human and mouse homologues of GPR65. Structure-activity relationship (SAR) studies revealed an analogue of BRD2813 with increased potency, BRD8716. BRD2813 and related potent analogues enable further study of the role of GPR65 in lysosomal function, autophagy, and inflammation in both cellular and in vivo models, and have already provided important information that could aid the future development of novel therapeutics for Inflammatory Bowel Disease.

Books / Online / Dissertations & Theses

English

January 29, 2018

Thesis (Ph.D.)--Yale University, 2017.

Yale University.