Investigating the role of fatty acid metabolism in melanocyte differentiation: Insights into the metabolic requirements of development

研究脂肪酸代谢在黑素细胞分化中的作用：深入了解发育的代谢要求

• 批准号: 10385715
• 负责人: Eleanor May Johns
• 金额: $ 4.68万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10385715
• 关键词: Adipocytes; Affect; Apoptosis; Binding Proteins; Brightfield Microscopy; CRISPR/Cas technology; Cell Lineage; Cell Proliferation; Cell membrane; Cells; Chemicals; Communication; Data; Defect; Dendrites; Dendritic Cells; Development; Disease; Embryo; Embryonic Development; Environment; Enzymes; Event; FABP3 gene; Family; Family member; Fatty Acids; Fluorescence Microscopy; Gene Expression; Genetic; Genetic Transcription; Hyperpigmentation; Image Analysis; Injections; Investigation; Knock-out; Label; Lead; Lecithin; Lipids; Measures; Melanoma Cell; Membrane; Metabolic; Metabolism; Methods; Mitochondria; Modeling; Morphology; Neural Crest; Nonesterified Fatty Acids; Oncogenic; Pathway interactions; Pattern; Pattern Formation; Phenotype; Phosphatidylethanolamine; Phospholipids; Pigmentation Disorders; Pigmentation physiologic function; Pigments; Play; Process; Production; Proliferating; Protein Family; Proteins; Role; Signal Transduction; Skin; Skin Cancer; Skin Pigmentation; Source; Supporting Cell; System; Testing; Transgenic Organisms; UV protection; Variant; Work; Zebrafish; base; cell motility; cell type; chemical genetics; experimental study; fatty acid metabolism; fatty acid oxidation; fatty acid transport; fatty acid-transport protein; genetic manipulation; human pluripotent stem cell; in vitro Model; in vivo; in vivo Model; insight; keratinocyte; lipid metabolism; melanoblast; melanocyte; melanoma; migration; novel; oxidation; promoter; sensor; tumor microenvironment; ultraviolet damage; uptake

PROJECT SUMMARY Melanocytes are a key cell lineage in the skin and are required for skin pigmentation. Defects in melanocyte development and function can lead to pigmentation disorders causing hypo or hyperpigmentation as well as one of the most deadly forms of skin cancer, melanoma. Many of the transcriptional and signaling events which cause these diseases have been explored, however, there has been little investigation into the role of the metabolic environment and intracellular metabolic state of the melanocyte during development. Recent work from our lab has demonstrated that during melanoma progression, adipocytes in the tumor microenvironment can transfer fatty acids to the melanoma cells, promoting invasion. This raises the question of whether fatty acid uptake plays a role in normal melanocytes. Our preliminary data suggests that melanocytes require extrinsic uptake of fatty acids through Fatty Acid Transport Proteins (FATPs) for proper differentiation. In Aim 1, we will investigate the relative importance of fatty acid uptake and de novo fatty acid synthesis in the melanocyte. We will compare multiple mechanisms of fatty acid uptake as well as fatty acid synthesis to determine which play the most important role in melanocyte development and pattern formation. We will also dissect in a stage specific manner when these pathways are required by the melanocyte during differentiation using genetic perturbations. We hypothesize that the importance of fatty acids in the melanocyte reflects certain lineage specific needs, including high levels of fatty acids to support phospholipid synthesis and dendrite formation. In Aim 2 we will examine the importance of fatty acids as building blocks for phospholipid synthesis relative to a requirement for fatty acid breakdown through b-oxidation. There is evidence for the importance of phospholipid synthesis in other dendritic cell types. Because melanocytes also form extensive dendrites which are critical for proper pigmentation we will dissect the importance major phospholipid synthesis pathways to the melanocyte in a stage specific manner. Conversely, melanocytes might break down fatty acids as a source of ATP to support cell proliferation and migration. We will also determine the role b-oxidation in melanocyte development by targeting key proteins in this pathway in a stage specific manner. We will assess the effects of these genetic perturbations on energy production and melanocyte development. To perform these studies we will primarily rely on zebrafish as an in vivo model of melanocyte development. We will use stage specific promoters for the neural crest, melanoblast, and melanocyte to generate stable germline transgenics. Using a combination of genetic methods and image analysis we will investigate the role of fatty acid and lipid metabolism in melanocyte development. When appropriate, we will also apply an in vitro model of melanocyte development, based on the differentiation of melanocytes from human pluripotent stem cells These combined experiments will lead to novel discoveries regarding the role of metabolism in melanocyte development, which could increase our understanding and treatment of pigmentation diseases.

项目摘要 黑素细胞是皮肤中的钥匙细胞谱系，是皮肤色素沉着所必需的。黑素细胞缺陷 开发和功能会导致色素沉着障碍，导致缺乏色调或色素沉着以及一种 皮肤癌最致命形式的黑色素瘤。引起的许多转录和信号事件 但是，已经探讨了这些疾病，但是，对代谢的作用几乎没有调查 发育过程中黑素细胞的环境和细胞内代谢状态。我们实验室的最新工作 已经证明，在黑色素瘤进展过程中，肿瘤微环境中的脂肪细胞可以转移 黑色素瘤细胞的脂肪酸，促进侵袭。这提出了一个问题，即脂肪酸吸收是否会发挥作用 在正常黑素细胞中的作用。我们的初步数据表明黑素细胞需要外部脂肪吸收 通过脂肪酸转运蛋白（脂肪）的酸以进行适当的分化。在AIM 1中，我们将调查 黑素细胞中脂肪酸摄取和从头脂肪酸合成的相对重要性。我们将 比较脂肪酸摄取的多种机制以及脂肪酸的合成，以确定哪个玩具 在黑素细胞发育和模式形成中最重要的作用。我们还将在特定阶段进行剖析 当黑素细胞使用遗传扰动分化时，黑素细胞需要这些途径时。 我们假设脂肪酸在黑素细胞中的重要性反映了某些特定的谱系需求， 包括高水平的脂肪酸，以支持磷脂合成和树突形成。在目标2中，我们将 研究脂肪酸作为磷脂合成的基础的重要性 通过B氧化对脂肪酸分解的要求。有证据表明 其他树突状细胞类型中的磷脂合成。因为黑素细胞也形成了广泛的树突 对于适当的色素沉着至关重要，我们将阐明重要的主要磷脂合成途径 黑素细胞以特定阶段的方式。相反，黑素细胞可能将脂肪酸分解为 ATP支持细胞增殖和迁移。我们还将确定黑素细胞中B-氧化的作用 通过以特定阶段的方式靶向该途径中的关键蛋白来开发。我们将评估 这些对能源生产和黑素细胞发育的遗传扰动。为了进行这些研究，我们 将主要依靠斑马鱼作为黑素开发的体内模型。我们将使用特定阶段 神经rest，黑素细胞和黑素细胞的启动子产生稳定的种系转基因。使用 遗传方法和图像分析的结合我们将研究脂肪酸和脂质代谢的作用 在黑素细胞发育中。在适当的时候，我们还将采用黑色素细胞发育的体外模型， 基于黑素细胞与人多能干细胞的分化，这些联合实验将 导致有关代谢在黑素细胞发育中的作用的新发现，这可能会增加我们 了解和治疗色素沉重疾病。