Chemical Genetic Analysis of Vertebrate Development Classic developmental genetics, involving large-scale mutagenesis screens, have shed much light on one of the most profound fundamental questions in biology: mechanistic insights into how a fertilized egg develops in an animal. Yet, because much of vertebrate development is inaccessible to traditional genetics, new approaches and tools are needed. As a complement to forward genetic analysis, we propose a large-scale chemical genetic analysis of early pattern formation in zebrafish embryos. Chemical genetic analysis, which involves the discovery and use of chemical probes, is a uniquely powerful approach to study vertebrate development. In a manner analogous to classic forward mutagenesis screens, a high-throughput chemical screen is conducted in zebrafish for small molecules that specifically modulate early embryonic development. Instead of mapping and cloning the responsible genes, the follow-up task involves identifying the pharmacological targets of the chemicals that elicit th prescribed phenotypic changes. As with forward genetic screens, the chemical genetic analysis can lead to the discovery of novel components or previously unanticipated signaling interactions involved. However importantly, since disturbances in developmental pathways are involved in the pathogenesis of many human illnesses, small molecules that selectively target them have significant translational and therapeutic potential. In the first specific aim, we will conduct a large-scale chemical screen for small molecules that specifically perturb early patterning in zebrafish embryos. As we have demonstrated, this is an innovative high content platform for discovering unique sets of highly selective modulators of developmental pathways. Next, we will employ chemical genetic analysis, which combines the methodologies of developmental biology, genetics, cell biology and pharmacology, to map the actions of small molecules discovered in Aim 1. This effort will be guided by the available molecular genetic information on early zebrafish embryogenesis. In addition, we propose a novel chemical genetic linkage analysis for target identification. We will combine lead optimization involving in vivo SAR (structure activity relationship) studies and target identification through large-scale drug target profiling. Finally, we propose an innovative platform for crowd sourcing of chemical genetic analyses and target identification efforts to overcome extant resource barriers that currently limi the impact of chemical genetics. In summary, the proposed study will discover valuable small molecules that selectively target key developmental pathways and use them to gain new insights on vertebrate development.
Disturbances in cell signaling pathways that regulate embryonic development lie at the heart of many human diseases, including those that affect adults such as cancers, and heart and muscle diseases. A better understanding of embryonic development, including chemical reagents to regulate it, will lead directly to important new therapies. Here, we will use zebrafish, a small tropical fish, to discover and test drug-like small molecules that control specific aspects of embryonic development, with the dual goal of studying fundamental biology and developing lead compounds for future therapeutics.
|Pulley, Jill M; Jerome, Rebecca N; Ogletree, Martin L et al. (2018) Motivation for Launching a Cancer Metastasis Inhibition (CMI) Program. Target Oncol 13:61-68|
|Durbin, Matthew D; Cadar, Adrian G; Chun, Young Wook et al. (2018) Investigating pediatric disorders with induced pluripotent stem cells. Pediatr Res 84:499-508|
|Pulley, Jill M; Shirey-Rice, Jana K; Lavieri, Robert R et al. (2017) Accelerating Precision Drug Development and Drug Repurposing by Leveraging Human Genetics. Assay Drug Dev Technol 15:113-119|
|Durbin, Matthew D; Cadar, Adrian G; Williams, Charles H et al. (2017) Hypoplastic Left Heart Syndrome Sequencing Reveals a Novel NOTCH1 Mutation in a Family with Single Ventricle Defects. Pediatr Cardiol 38:1232-1240|
|Williams, Charles H; Hong, Charles C (2016) Zebrafish small molecule screens: Taking the phenotypic plunge. Comput Struct Biotechnol J 14:350-356|
|Hempel, Jonathan E; Cadar, Adrian G; Hong, Charles C (2016) Development of thieno- and benzopyrimidinone inhibitors of the Hedgehog signaling pathway reveals PDE4-dependent and PDE4-independent mechanisms of action. Bioorg Med Chem Lett 26:1947-53|
|Jiramongkolchai, Pawina; Owens, Philip; Hong, Charles C (2016) Emerging roles of the bone morphogenetic protein pathway in cancer: potential therapeutic target for kinase inhibition. Biochem Soc Trans 44:1117-34|