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: 10642685
• 负责人: Eleanor May Johns
• 金额: $ 4.77万
• 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/10642685
• 关键词: Adipocytes; Affect; Apoptosis; Binding Proteins; 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; Genes; Genetic; Genetic Transcription; Genus Hippocampus; Hyperpigmentation; Image Analysis; Injections; Invaded; Investigation; Knock-out; Label; 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; Play; Process; Production; Proliferating; Protein Family; Protein Inhibition; Proteins; Role; Signal Transduction; Skin; Skin Cancer; Skin Pigmentation; Source; Supporting Cell; System; Testing; UV protection; Variant; Work; Zebrafish; cell motility; cell type; 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关键细胞谱系，并且是皮肤色素沉着所需的。黑素细胞缺陷 发育和功能可导致色素沉着障碍，引起色素沉着不足或色素沉着过度， 最致命的皮肤癌黑色素瘤许多转录和信号事件， 这些疾病已被探索，然而，很少有研究的作用，代谢 环境和细胞内的代谢状态的黑素细胞在发育过程中。我们实验室最近的工作 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， 脂肪酸对黑色素瘤细胞，促进入侵。这就提出了一个问题，脂肪酸的摄取是否起着 在正常黑色素细胞中的作用。我们的初步数据表明，黑素细胞需要外源性摄取脂肪酸， 脂肪酸通过脂肪酸转运蛋白（FATPs）进行适当的分化。在目标1中，我们将研究 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 比较脂肪酸摄取和脂肪酸合成的多种机制，以确定哪些发挥作用， 在黑素细胞发育和图案形成中起着最重要的作用。我们还将在特定的阶段进行解剖 当黑素细胞在分化过程中需要这些途径时，使用遗传扰动的方式。 我们假设脂肪酸在黑素细胞中的重要性反映了某些谱系的特定需求， 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， 检查脂肪酸作为磷脂合成基石的重要性 需要通过β-氧化分解脂肪酸。有证据表明， 其他类型树突状细胞的磷脂合成。因为黑素细胞也形成广泛的树突， 是至关重要的适当色素沉着，我们将解剖的重要性，主要磷脂合成途径， 黑色素细胞的阶段特异性的方式。相反，黑色素细胞可能会分解脂肪酸作为一种来源， ATP支持细胞增殖和迁移。我们还将确定B-氧化在黑素细胞中的作用， 通过以阶段特异性方式靶向该途径中的关键蛋白质来促进发育。我们将评估 这些遗传干扰对能量产生和黑色素细胞发育的影响。为了进行这些研究，我们 将主要依靠斑马鱼作为黑素细胞发育的体内模型。我们将使用特定阶段 用于神经嵴、成黑素细胞和黑素细胞的启动子，以产生稳定的种系转基因。使用 结合遗传学方法和图像分析，我们将研究脂肪酸和脂质代谢的作用 在黑素细胞的发育中。在适当的时候，我们还将应用黑素细胞发育的体外模型， 基于从人类多能干细胞中分化出黑素细胞。 导致新的发现代谢在黑素细胞发育中的作用，这可能会增加我们的研究。 了解和治疗色素沉着疾病。