Delineate the regulatory network of lipid metabolism via SRS imaging-sorting

 描述（由申请人提供）：脂质分子不仅作为能量来源或物理屏障，还积极参与调节细胞信号传导、膜运输和转录网络。脂质代谢功能障碍有助于各种人类疾病的发展。脂肪酸是所有脂质分子的组成部分和前体，对细胞生理学和病理学有重要影响。饱和脂肪酸在非脂肪组织中的过量积累导致细胞毒性和非酒精性脂肪肝疾病、心肌病和II型糖尿病的发病机制;而不饱和脂肪酸毒性较低，甚至发挥保护作用。最近的研究表明，脂肪酸的脂毒性差异可能与其不同的效率，由于纳入脂滴（LD）。然而，控制不同脂肪酸分子之间LD掺入异质性的潜在分子机制仍然知之甚少。在我们最近的研究中，我们开发了一种新的化学成像策略，用于在活细胞和生物体中以亚细胞分辨率跟踪特定脂质分子的时空动态，并直接可视化饱和和不饱和脂肪酸在其掺入LD期间的差异。该提案旨在剖析调节不同脂肪酸分子之间LD掺入异质性的分子机制，并将联合收割机创新技术开发和详细的机制表征相结合。目标1将开发基于新型化学成像显微镜的高通量成像/分选微流体平台。目的2通过高通量正向遗传筛选，阐明调控脂肪酸代谢动力学的新基因和新途径。我们已经为这个项目与科学家建立了独特的合作关系。该研究的成功完成将为全组织显微筛选提供新的技术平台，并为脂肪酸代谢及其生理和病理功能提供新的见解。

 DESCRIPTION (provided by applicant): Lipid molecules act not only as energy resources or physical barriers, they are also actively involved in regulating cellular signaling, membrane trafficking, and the transcriptional network. Lipid metabolic dysfunction contributes to the development of various human diseases. Fatty acids are building blocks and precursors of all lipid molecules and have crucial impact on cell physiology and pathology. Excess accumulation of saturated fatty acids in non-adipose tissues leads to cytotoxicity and the pathogenesis of nonalcoholic fatty liver diseases, cardiomyopathy and type II diabetes mellitus; whereas unsaturated fatty acids are less toxic and even exert protective effects. Recent studies suggest that differences of fatty acids in lipotoxicity may be associated with their distinct efficiency duing the incorporation into lipid droplets (LDs). However the underlying molecular mechanisms governing this LD incorporation heterogeneity between different fatty acid molecules remain poorly understood. In our recent studies, we have developed a new chemical imaging strategy for tracking the spatiotemporal dynamics of specific lipid molecules at the subcellular resolution in living cells and organisms, and directly visualized the difference between saturated and unsaturated fatty acids during their incorporation into LDs. This proposal aims to dissect the molecular mechanisms that regulate LD incorporation heterogeneity between different fatty acid molecules, and will combine innovative technology development and detailed mechanistic characterization. Aim 1 will develop a high-throughput imaging/sorting microfluidic platform based the new chemical imaging microscopy. Aim 2 will elucidate new genes and pathways in regulating fatty acid metabolic dynamics through high-throughput forward genetic screens. We have established unique collaborations with scientists for this project. Successful accomplishment of the proposed studies will provide a new technology platform for whole-organism microscopic screening, and provide new insights into fatty acid metabolism and their physiological and pathological functions.