Transcription factors regulate distinct networks of genes in specific tissues, yet understanding how these networks are coordinated temporally and spatially to promote animal development and homeostasis is extremely challenging. Here, I propose to study how gene regulatory networks promote a developmental program (molting) using the simplicity of the model nematode C. elegans. I will test the hypothesis that nuclear hormone receptor regulation of core elements of the molting network are conserved between free-living and parasitic nematodes by comparing the C. elegans molt network to that of the human parasite, B. malayi. Since arriving at UCSF, I have been awarded both Terry Fox Foundation and Canadian Institutes of Health postdoctoral fellowships for work that explored how nuclear hormone receptors promote organ development and innate immunity in C. elegans. Two nuclear hormone receptors (NHR-23 and NHR-25) are key regulators of C. elegans molting, and I will investigate the roles of these transcription factors in the molting network. Hee I propose to: i) define the components of the C. elegans molting network;ii) define functional modules in the C. elegans molt, tissues of action and spatial regulation;and iii) extend my C. elegans findings into the human parasite, B. malayi.
These aims will be achieved through a combination of ChIP-seq, proteomics, data mining, quantitative genetic analysis, genome editing, and standard C. elegans techniques. Combining the focused genetic approach with an unbiased systems approach will deliver genes/proteins that change during the molt, identify direct and indirect targets of NHR-23 and NHR-25, and determine the factors involved in temporal and spatial regulation of select groups of molting genes. In the long-term, I will investigate the mechanisms of transcriptional coordination in an intact animal. To achieve these aims, I will acquire new training from my co-mentors Dr. Keith Yamamoto (ChIP-seq) and Dr. Nevan Krogan (bioinformatics analysis/ programming, mass spectrometry, quantitative genetic analysis), and from my Advisory Group members Dr. Alison Frand (molting physiology) and Dr. Judy Sakanari (B. malayi culture/physiology). My rigorous career plan, and outstanding and comprehensive mentoring team will put me in a strong position to secure an independent position in a U.S. academic institution. The renowned community spirit of UCSF combined with its rich technical, scientific, and professional development resources will provide an ideal incubator for me to develop this project and prepare for independence. A K99/R00 grant will allow me to build on my expertise in C. elegans nuclear hormone receptors and gene expression, using both unbiased systems approaches and focused genetics/cell biology, to study developmental gene regulatory networks in the context of an entire animal. This work will provide insight into the structure and evolution of transcriptional networks, and extension of this knowledge to B. malayi will pave the way for investigations into molting as an intervention point in treating parasitic nematodes, which burden 2.9 billion people globally.
2.9 billion people worldwide are burdened with parasitic nematodes, yet we only have four drugs and they are losing their effectiveness. These parasitic nematodes cause significant pain, developmental and mental delay and mire developing countries in poverty, hampering the global economy. I will determine the mechanisms of a process unique to nematodes (molting) using a non-parasitic model nematode and extend my findings into the parasite, in an effort to provoke development of new, innovative drugs.