Spatial metabolomics with subcellular resolution to identify therapeutic targets

摘要 细胞死亡是一个重要的生理过程。癌细胞对治疗药物的抗药性是一个重要的 是癌症治疗成功的障碍，也是癌症复发的主要因素。激活新的细胞死亡 途径将使耐药细胞对化疗重新敏感。铁下垂是一种非凋亡性细胞死亡。 当细胞中的脂质经历铁依赖的过氧化时被激活。我们的首要目标是调查 脂代谢通过调节细胞培养中铁下垂和体内药物模型而导致细胞死亡。 耐药癌症。这些结果将为铁性上睑下垂及其相关疾病的分子化学研究提供基础资料。 通过开发具有单细胞分辨率的多组学方法，在疾病病理学中发挥作用。癌症等疾病， 脓毒症、先兆子痫、糖尿病、心血管疾病和神经退行性疾病与血脂相关 脂代谢紊乱。脂质的分布是不均匀的，它们的化学修饰，如 作为脂质过氧化，对疾病的发生和发展具有潜在的关键作用。然而，准确的关系是 脂类分布及其化学修饰和疾病病理之间的关系尚不完全清楚。这 该项目调查了体外和组织中脂质分布和脂质过氧化的变化，以提供 通过实验对它们的分子化学及其在疾病病理学中的作用的基本见解 本课题组开发的基于空间化学成像的方法。我们的方法使用拉曼成像技术 提供有关定义的细胞隔间中脂质的空间信息，而不是散装或分级的 采用液-质联用(LC-MS)对提取的脂质进行检测。这项建议 建立在我们之前成功地对细胞和组织中的脂肪分布进行空间成像的基础上。这个 这一努力背后的假设是：(A)在脂质分布和他们的 化学修饰，以及(B)这些异质性可以与疾病的病理相关联。在……里面 具体地说，我们将：1)研究铁下垂对药物脂代谢和铁代谢的影响。 2D培养中的耐药细胞系；2)3D细胞中铁下垂时脂质的空间定位和分布 培养；3)体内检测铁性下垂及其相关机制。项目成果将有所改善 我们对分子机制、疾病表型和疾病进展的理解有助于更好地 治疗策略。

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.