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性是一个重要的因素。 它是成功治疗癌症的障碍和癌症复发的主要因素。激活新的细胞死亡 这些通路会使耐药细胞对化疗重新敏感。铁凋亡是一种非凋亡性细胞死亡 当细胞中的脂质经历铁依赖性过氧化时被激活。我们的首要目标是调查 脂质代谢驱动的细胞死亡，通过在细胞培养和体内模型的药物， 耐药性癌症这些结果将为铁凋亡的分子化学及其治疗提供基本的见解。 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。像癌症， 败血症、先兆子痫、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。脂质分布是不均匀的，它们的化学修饰，如 如脂质过氧化，对疾病的发生和发展可能至关重要。然而，精确的关系 脂质分布及其化学修饰与疾病病理学之间的关系尚未完全了解。这 该项目研究了体外和组织中脂质分布和脂质过氧化的变化， 基本的见解，他们的分子化学及其在疾病病理学中的作用，通过使用实验 我们小组基于空间化学成像开发的方法。我们的方法使用拉曼成像， 提供了关于限定的细胞区室中脂质的空间信息， 通过液相色谱-质谱法（LC-MS）提供提取的脂质的检查。这项建议 建立在我们先前成功地对细胞和组织中的脂质分布进行空间成像的基础上。的 这一努力的基础假设是：（a）脂质分布及其 化学修饰，和（B）这些异质性可以与疾病的病理学相关。在 本研究主要包括以下几个方面：1）探讨铁缺乏症对药物代谢的影响; 2）在3D细胞中进行铁凋亡期间脂质的空间映射和分析 3）检测体内铁凋亡并确定其相关机制。项目成果将得到改善 我们对分子机制、疾病表型和疾病进展的理解， 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.