The principal aim of this proposal is to elucidate the structures and activation and regulation mechanisms of the multi-protein complex inflammasomes, and to develop tool compounds that perturb the assembly of the inflammasomes with potentials for future therapeutic interventions. Dr. Qian Yin is currently a research fellow in Boston Children's Hospital and Harvard Medical School with an immediate goal of completing one-year mentored research before establishing herself in a tenure-track academic position. Dr. Yin's long-term goals include independent researches to understand human innate immune and inflammatory systems and their interaction with pathogens and environmental and intrinsic stresses, with the hope of finding amenable therapeutic targets and effective means for immune and inflammatory diseases. Dr. Yin is a well-trained structural biologist specialized in innate immune and inflammatory signaling. The project will be conducted at Boston Children's Hospital in the rich research community of Longwood Medical Area of Boston, with all facilities and resources from neighboring Harvard Medical School available to the candidate. Dr. Yin will be mentored by Dr. Hao Wu in structural and biophysical approaches to elucidate inflammasome structures and co-mentored by Dr. Timothy Mitchison in development of high throughput assays and tool compounds. During the one-year mentored period, Dr. Yin will learn new techniques such as electron microscopy and high throughput screening under the guidance of the mentors, and develop professional skills by attending classes, seminars, and workshops. After that Dr. Yin plans to make the transition to independence with the assistance of the proposed awards. Inflammasomes are multi-protein complexes that launch the first-line inflammatory responses. During the mentored phase Dr. Yin proposes to reconstruct high-resolution electron microscopy (EM) structure of the AIM2 inflammasome, with X-ray crystallography providing detailed interfacial interactions (Aim 1). This period will also see Dr. Yin clarify the working mechanism of p202, a natural AIM2 inflammasome inhibitor. The proposed work builds heavily on the preliminary EM data of in vitro reconstitution of ASC PYD domain, a common adaptor in most inflammasomes. For the independent phase, Dr. Yin will 1) investigate the assembly mechanisms of NLRP3 inflammasome and elucidate the differential requirements among various inflammasomes (Aim 2), and 2) assay structure-based stabilized helical peptides and screen chemical libraries for small molecules that can perturb inflammasome assembly, in the hope to find druggable interfaces and tool compounds with potential therapeutic values (Aim 3). RELEVANCE: Inflammation is a self-protection process that needs to be tightly regulated. Understanding inflammasome activation and regulation will provide new avenues for treatment of many metabolic diseases, e.g. diabetes and various cancers, as well as hereditary autoinflammatory diseases.