Spatial metabolomics with subcellular resolution to identify therapeutic targets

具有亚细胞分辨率的空间代谢组学以确定治疗靶点

• 批准号: 10714487
• 负责人: Manas Ranjan Gartia
• 金额: $ 37.5万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-07 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10714487
• 关键词: Cardiovascular Diseases; Cell Culture Techniques; Cell Death; Cell Line; Cells; Chemicals; Chemistry; Diabetes Mellitus; Disease; Disease Progression; Drug Sensitization; Drug resistance; Goals; Heterogeneity; Image; In Vitro; Iron; Lipid Peroxidation; Lipids; Malignant Neoplasms; Maps; Methods; Modification; Molecular; Neurodegenerative Disorders; Onset of illness; Outcome; Pathology; Pathway interactions; Pharmaceutical Preparations; Physiological Processes; Pre-Eclampsia; Raman Spectrum Analysis; Recurrent Malignant Neoplasm; Resistance; Resolution; Role; Sepsis; Techniques; Therapeutic; Tissues; cancer cell; cancer recurrence; cancer therapy; chemotherapy; disease phenotype; improved; in vivo; in vivo Model; insight; iron metabolism; lipid metabolism; lipidomics; liquid chromatography mass spectrometry; metabolomics; multiple omics; novel; peroxidation; refractory cancer; success; therapeutic target; three dimensional cell culture

Abstract Cell death is a crucial physiological process. The resistance of cancer cells to therapeutic drugs is a significant barrier to successful cancer treatment and the primary factor in cancer recurrence. Activation of novel cell death pathways would resensitize drug-resistant cells to chemotherapy. Ferroptosis is a nonapoptotic cell death activated when the lipid in the cell undergoes iron-dependent peroxidation. Our overarching goal is to investigate lipid metabolism-driven cell death through the modulation of ferroptosis in cell culture and in vivo models of drug- resistant cancer. The results will provide fundamental insights into the molecular chemistry of ferroptosis and its role in disease pathology by developing a multi-omics approach with single-cell resolution. Diseases like cancer, sepsis, pre-eclampsia, diabetes, cardiovascular disease, and neurodegenerative illnesses correlate with lipids and lipid metabolism dysregulation. Lipid distributions are heterogeneous, and their chemical modifications, such as lipid peroxidation, are potentially crucial for disease onset and progression. However, the precise relationship between lipid distribution and their chemical modification and disease pathology is not fully understood. This project investigates changes in lipid distribution and lipid peroxidation both in vitro and in tissues to provide fundamental insights into their molecular chemistry and its role in disease pathology by using the experimental methods developed in our group based on spatial chemical imaging. Our approach uses Raman imaging to provide spatial information about lipids in the defined cellular compartments in contrast to the bulk or fractionated examinations of extracted lipids provided by liquid chromatography-mass spectrometry (LC-MS). This proposal builds upon our prior success in performing spatial imaging of lipid distribution in cells and tissues. The hypotheses underlying this effort are that (a) there are significant heterogeneities in lipid distributions and their chemical modifications, and (b) these heterogeneities can be correlated to the pathology of the disease. In particular, we will: 1) Investigate the effect of ferroptosis on the lipid metabolism and iron metabolism of drug- resistant cell lines in 2D culture; 2) Perform spatial mapping and profiling of lipids during ferroptosis in 3D cell culture; 3) Detect ferroptosis and identify its associated mechanism in vivo. The project outcomes will improve our understanding of the molecular mechanism, disease phenotype, and disease progression leading to better therapeutic strategies.

抽象的 细胞死亡是一个至关重要的生理过程。癌细胞对治疗药物的抗性是重要的 成功癌症治疗的障碍和癌症复发的主要因素。新细胞死亡的激活 途径将对化学疗法产生抗药性细胞的敏感性。铁凋亡是一种非凋亡细胞死亡 当细胞中的脂质经历铁依赖性过氧化时，被激活。我们的总体目标是调查 通过调节细胞培养和药物的体内模型，通过脂质代谢驱动的细胞死亡。 抗性癌症。结果将提供对铁凋亡的分子化学及其的基本见解 通过开发单细胞分辨率的多词方法在疾病病理学中的作用。癌症等疾病， 败血症，前球，糖尿病，心血管疾病和神经退行性疾病与脂质相关 和脂质代谢失调。脂质分布是异质的，它们的化学修饰是这样的 作为脂质过氧化，对疾病的发作和进展至关重要。但是，确切的关系 脂质分布与其化学修饰和疾病病理学之间尚不完全了解。这 项目研究了体外和组织中脂质分布和脂质过氧化的变化，以提供 通过使用实验，对其分子化学及其在疾病病理中的作用的基本见解 基于空间化学成像在我们组中开发的方法。我们的方法使用拉曼成像 与散装或分数相比，在定义的细胞室中提供有关脂质的空间信息 通过液相色谱 - 质谱法（LC-MS）提供的提取的脂质的检查。这个建议 建立在我们先前成功地对细胞和组织中脂质分布进行空间成像的成功。这 这项工作为基础的假设是（a）脂质分布及其它们的异质性很大 化学修饰，（b）这些异质性可以与疾病的病理相关。在 特别是，我们将：1）研究铁铁作用对药物的脂质代谢和铁代谢的影响 2D培养中的抗性细胞系； 2）在3D细胞中的脂质吞噬过程中进行脂质的空间映射和分析 文化; 3）检测铁凋亡并在体内鉴定其相关的机制。项目成果将改善 我们对分子机制，疾病表型和疾病进展的理解导致更好 治疗策略。