Cancer Biology Research Test-Bed Unit 2: Effects of cell-intrinsic and cell-extrinsic variations in lipid metabolism on metastasis patterns

癌症生物学研究试验台单元 2：脂质代谢的细胞内在和细胞外在变化对转移模式的影响

• 批准号: 10374653
• 负责人: SEAN J MORRISON
• 金额: $ 33.77万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-24 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10374653
• 关键词: Address; Affect; Antioxidants; Beds; Biological Assay; Blood; Brain; Cancer Biology; Cancer Control; Cell Death; Cell Proliferation; Cell Survival; Cells; Clinical Trials; Data; Distant; Environment; Exhibits; Exposure to; Flow Cytometry; Hydrogen; Image; Immunocompetent; Immunocompromised Host; Impairment; In Situ; Intervention; Kidney; Lipid Peroxides; Lipids; Liver; Location; Lung; Lymph; Melanoma Cell; Membrane; Metabolic; Mitochondria; Monounsaturated Fatty Acids; Mus; Mutation; Neoplasm Metastasis; Oleic Acids; Organ; Oxidative Stress; Patients; Pattern; Phospholipids; Polyunsaturated Fatty Acids; Process; Proliferating; Property; Reactive Oxygen Species; Research; Resistance; Resolution; Site; Testing; Tissues; Variant; Xenograft procedure; cancer cell; cellular imaging; dietary; experience; in vivo; lipid metabolism; lipidomics; lymphatic vessel; melanoma; oxidation; response; technology development; tumor progression

Project Summary Metastasis is a highly inefficient process in which few disseminating cancer cells survive. We discovered that melanoma metastasis is limited by oxidative stress. Reactive Oxygen Species (ROS) increase dramatically in melanoma cells as they metastasize through the blood. The rare cells that survive undergo reversible metabolic changes that confer oxidative stress resistance. Consistent with this, large clinical trials found that patients administered anti-oxidants were more likely to die of cancer than control patients. Cancer cells, including melanoma, often metastasize regionally through lymphatic vessels before metastasizing systemically through the blood. We recently discovered that melanoma cells in lymph experience less oxidative stress and form more metastases than melanoma cells in blood. This was true of patient-derived melanomas growing in immunocompromised mice as well as mouse melanomas growing in immunocompetent mice. The oxidative stress kills melanoma cells in the blood by inducing ferroptosis, a form of cell death marked by the accumulation of lipid peroxides. One of the ways in which lymph protects from ferroptosis is by having high levels of the monounsaturated fatty acid (MUFA), oleic acid, which protects cells from lipid oxidation by reducing the abundance of polyunsaturated fatty acids (PUFAs) in membrane phospholipids. The more abundant PUFAs are in membrane phospholipids, the more sensitive cells are to ferroptosis. Melanoma cells are thus exposed to different lipid environments in different locations as they metastasize and these cell- extrinsic changes influence their survival. There are also cell-intrinsic differences among melanomas that influence their response to these environments: melanomas from different patients differ in the sites to which they metastasize upon xenografting. We hypothesize that these differences in metastasis patterns result from cell-intrinsic differences in lipid metabolism that influence their sensitivity to ferroptosis in response to the lipid environments they encounter as they metastasize. Nonetheless, we have never been able to image the fates of metastasizing melanoma cells in vivo, limiting our understanding of how oxidative stress affects these cells. A critical barrier is the efficient imaging of entire organs to identify rare melanoma cells at the earliest stages of metastasis. The lack of subcellular resolution in whole organs also impairs our ability to explore the ways in which oxidative stress influences cell survival and proliferation. Both of these limitations will be addressed by the Technology Development Unit (TDU) of this consortium, which is developing the ability to perform high throughput imaging of whole, cleared organs to assess the survival, proliferation, and localization of rare melanoma cells after metastasis. We will validate these imaging data by flow cytometry. By combining single cell imaging (Aim 1) with metabolic assays, we will characterize cell-extrinsic (Aim 2) and cell-intrinsic (Aims 1, 3) mechanisms that regulate the earliest stages of metastasis.

项目摘要 转移是一个高效的过程，其中很少有传播癌细胞的生存。我们发现了这一点 黑色素瘤转移受到氧化应激的限制。活性氧（ROS）在 黑色素瘤细胞通过血液转移。生存的稀有细胞可逆 代谢变化，赋予氧化应激抗性。与此相一致，大型临床试验发现 与对照患者相比，服用抗氧化剂的患者更有可能死于癌症。癌细胞， 包括黑色素瘤在内，通常会通过淋巴管进行区域转移，然后全身转移 通过血液。我们最近发现，淋巴中的黑色素瘤细胞经历较少的氧化应激和 在血液中比黑色素瘤细胞形成更多的转移。在患者衍生的黑色素瘤中生长在 免疫功能低下的小鼠以及在免疫能力小鼠中生长的小鼠黑色素瘤。氧化 压力通过诱导铁铁病杀死血液中的黑色素瘤细胞，这是一种细胞死亡形式 脂质过氧化物的积累。淋巴保护免受铁铁作用的方法之一是高 单不饱和脂肪酸（MUFA）的水平，油酸，可通过保护细胞免受脂质氧化的影响 减少膜磷脂中多不饱和脂肪酸（PUFAS）的丰度。更多 大量的pufas在膜磷脂中，更敏感的细胞是屈服。黑色素瘤细胞 因此，在转移时会暴露于不同位置的不同脂质环境，这些细胞 - 外在变化会影响其生存。黑色素瘤之间也存在细胞中性差异 影响他们对这些环境的反应：来自不同患者的黑色素瘤在所在地不同 它们在异武施thing上转移。我们假设转移模式的这些差异是由 脂质代谢的细胞中性差异会影响其对脂质的敏感性的敏感性 他们转移时遇到的环境。尽管如此，我们从来没有能够为命运做出成像 在体内转移黑色素瘤细胞的，限制了我们对氧化应激如何影响这些细胞的理解。 关键障碍是整个器官的有效成像，以鉴定最早的稀有黑素瘤细胞 转移。整个器官缺乏亚细胞分辨率也损害了我们探索方式的能力 氧化应激影响细胞的存活和增殖。这两个限制将由 该财团的技术开发单元（TDU），正在发展高度的能力 整体，清除器官的吞吐量成像，以评估罕见的生存，增殖和定位 转移后黑色素瘤细胞。我们将通过流式细胞仪验证这些成像数据。通过组合单 细胞成像（AIM 1）带有代谢测定法，我们将表征细胞超支（AIM 2）和细胞中心（AIMS 1， 3）调节转移最早阶段的机制。