A genetically-encoded sensor for imaging intracellular fatty acids

用于细胞内脂肪酸成像的基因编码传感器

• 批准号: 8251128
• 负责人: James G Granneman
• 金额: $ 22.8万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-05 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8251128
• 关键词: Biological; Cardiovascular Diseases; Caveolae; Cell physiology; Cells; Cellular biology; Development; Diabetes Mellitus; Disease; Evolution; Fatty Acids; Fluorescence; Fluorescence Resonance Energy Transfer; Functional disorder; Health; Image; Investigation; Lead; Life; Lipid Mobilization; Lipids; Mammalian Cell; Metabolism; Mitochondria; Obesity; Resolution; Site; Work; base; design; detector; insight; lipid transport; novel; optical sensor; oxidation; prototype; public health relevance; sensor; uptake

DESCRIPTION (provided by applicant): The storage and mobilization of lipid are fundamental cellular processes that are conserved throughout metazoan evolution and are present in virtually all mammalian cells. Intracellular FFA (FFAi) are normally kept under tight control since excessive FFAi are toxic. Indeed, excessive accumulation of intracellular FFA and FFA metabolites leads to cellular dysfunction, termed 'lipotoxicity', which is thought to be a major means by which obesity contributes to diabetes and cardiovascular disease. Thus, a mechanistic understanding of how cells assimilate, mobilize and channel FFA is an important biological question with broad implications for health and disease. The uptake and mobilization of FFA within cells appear to have important temporal and spatial domains. Nonetheless, investigation of the "when and where" FFA metabolism occurs within live cells has not been possible because intracellular FFA levels cannot presently be imaged. Therefore, the aim of the R21 application is to develop and implement an optical sensor of FFAi having high spatial and temporal resolution for biological applications. This sensor is designed to be used as a general cytosolic sensor, or can be specifically targeted to proposed subcellular sites of FFA uptake, mobilization and oxidation. We anticipate that a validated sensor will have wide applications and will lead to new insights into biology of cellular lipid trafficking. PUBLIC HEALTH RELEVANCE: The storage and mobilization of lipid are fundamental cellular processes that are present in virtually all mammalian cells. Intracellular FFA (FFAi) are normally kept under tight control and since excessive FFAi are toxic and can contribute to obesity-related diabetes and cardiovascular disease. Thus, a mechanistic understanding of how cells assimilate, mobilize and channel FFA is an important biological question with broad implications for health and disease. Evidence indicates that the uptake and mobilization of FFA within cells has important temporal and spatial domains, however, analysis of these dynamics has not been difficult because intracellular FFA levels cannot presently be imaged. Therefore, the aim of the R21 application is to develop and implement an optical sensor of intracellular FFA with high spatial and temporal resolution for biological applications. This sensor can be used as a general cytosolic sensor, or can be specifically targeted to key subcellular sites of FFA uptake, mobilization and oxidation. We anticipate that a validated sensor will have wide applications and will lead to new insights into biology of cellular lipid trafficking.

描述（由申请人提供）：脂质的储存和动员是在后生动物进化过程中保守的基本细胞过程，几乎存在于所有哺乳动物细胞中。细胞内FFA（FFAi）通常保持在严格控制下，因为过量的FFAi是有毒的。 事实上，细胞内FFA和FFA代谢物的过度积累导致细胞功能障碍，称为“脂毒性”，这被认为是肥胖导致糖尿病和心血管疾病的主要手段。因此，了解细胞如何吸收、动员和引导FFA是一个重要的生物学问题，对健康和疾病具有广泛的影响。 FFA在细胞内的摄取和动员似乎具有重要的时间和空间域。 尽管如此，活细胞内FFA代谢发生的“时间和地点”的调查一直是不可能的，因为细胞内FFA水平目前无法成像。 因此，R21申请的目的是开发和实现具有高空间和时间分辨率的FFA1光学传感器，用于生物应用。 该传感器被设计为用作一般的胞质传感器，或者可以专门针对FFA摄取、动员和氧化的亚细胞位点。 我们预计，经过验证的传感器将有广泛的应用，并将导致新的见解生物学的细胞脂质贩运。 公共卫生相关性：脂质的储存和动员是基本的细胞过程，几乎存在于所有哺乳动物细胞中。细胞内FFA（FFAi）通常保持在严格的控制下，因为过量的FFAi是有毒的，可以导致肥胖相关的糖尿病和心血管疾病。因此，了解细胞如何吸收、动员和引导FFA是一个重要的生物学问题，对健康和疾病具有广泛的影响。有证据表明，细胞内FFA的摄取和动员具有重要的时间和空间域，然而，分析这些动力学并不困难，因为细胞内FFA水平目前无法成像。 因此，R21应用的目的是开发和实现用于生物应用的具有高空间和时间分辨率的细胞内FFA的光学传感器。 该传感器可用作一般的胞质传感器，或者可特异性地靶向FFA摄取、动员和氧化的关键亚细胞位点。 我们预计，经过验证的传感器将有广泛的应用，并将导致新的见解生物学的细胞脂质贩运。