Investigating the Role of SLC45A4 in GABA Metabolism

研究 SLC45A4 在 GABA 代谢中的作用

• 批准号: 10635402
• 负责人: Xiaoyang Su
• 金额: $ 32.19万
• 依托单位: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10635402
• 关键词: Affect; Amino Acids; Antibodies; Arginine; Biochemical; Biochemistry; Carrier Proteins; Cations; Cell Line; Cell Proliferation; Cell membrane; Cell physiology; Cells; Central Nervous System; Chemicals; Coupled; Cultured Cells; Cytosol; Data; Data Set; Disease; Drug Targeting; Environment; Family; Family member; Genes; Glucose; Glucose Transporter; Goals; Human; Immunofluorescence Immunologic; Investigation; Ions; Knock-out; Knowledge; Label; Mass Spectrum Analysis; Measures; Mediating; Membrane; Membrane Transport Proteins; Mendelian disorder; Metabolic; Metabolism; Mitochondria; Neuroglia; Neurons; Neurotransmitters; Normal Cell; Organelles; Ornithine; Orphan; Paraquat; Peripheral; Plants; Production; Property; Proteins; Protons; Putrescine; Regulation; Resolution; Role; Sequence Homology; Spermidine; Spermine; Sucrose; Techniques; Testing; Tissues; Work; cancer cell; cell growth; cytotoxicity; drug development; extracellular; gamma-Aminobutyric Acid; member; metabolomics; migration; mitochondrial membrane; multiple omics; new therapeutic target; novel; novel therapeutics; ornithine transporter; overexpression; preference; programs; small molecule; solute; stable cell line; stable isotope; transcriptomics; uptake

ABSTRACT Solute carrier (SLC) proteins are membrane transporters that govern the cross-membrane exchanges of glucose, amino acids, inorganic ions, and other small molecule metabolites. Many SLC genes have been shown to be causes of Mendelian diseases in humans, and a number of SLC transporters are important drug targets. However, due to myriad technical difficulties, a large fraction of SLC family members are still orphan transporters without known substrates, which represents both a significant knowledge gap and a huge opportunity for new drug development. In order to systematically study the biochemical functions of the orphan SLC transporters, a computational workflow was developed in our lab, which combines public transcriptomic and metabolomic datasets to uncover the metabolic function of SLC transporters. Using this association analysis, an uncharacterized gene, SLC45A4, was identified to be the single greatest determinant of γ-Aminobutyric acid (GABA) levels in human cancer cells. GABA, which is mostly known as an inhibitory neurotransmitter in the mammalian central nervous system, functions in peripheral tissues to regulate cell proliferation, differentiation, and migration. Given the importance of these functions in both cancer and normal cells, it is essential to understand how SLC45A4 functions in the context of GABA metabolism in non-neuronal cells. Using stable isotope tracing, it was found that SLC45A4 increases cellular GABA levels not by promoting GABA uptake but by facilitating GABA de novo synthesis, suggesting an entirely new mechanism of the regulation of GABA synthesis. Therefore, it is hypothesized that SLC45A4 encodes a subcellular ornithine transporter that supports GABA production. This program will move forward in three directions: Aim 1, determining the biochemical mechanism by which SLC45A4 promotes cellular GABA production; Aim 2, determining the subcellular compartmentalization of SLC45A4 transporter and GABA synthesis; and Aim 3, determining substrate preference and enzymatic properties of SLC45A4. The overall goal is to determine how SLC45A4 regulates de novo GABA synthesis, which contributes to the fundamental knowledge of human biochemistry and may provide new therapeutics targeting GABA metabolism.

抽象的 溶质载体（SLC）蛋白是膜转运蛋白，它控制着葡萄糖的跨膜交换， 氨基酸，无机离子和其他小分子代谢产物。许多SLC基因已显示为 人类孟德尔疾病的原因，许多SLC转运蛋白是重要的药物靶标。 但是，由于无数的技术困难，很大一部分SLC家庭成员仍然是孤儿运输公司 没有已知的底物，这既代表了重大的知识差距，又代表了新的机会 药物开发。为了系统地研究孤儿SLC转运蛋白的生化功能 计算工作流是在我们的实验室中开发的，该实验室结合了公共转录组和代谢组。 数据集以发现SLC转运蛋白的代谢功能。使用此关联分析， 未表征的基因SLC45A4被认为是γ-氨基丁酸的最大决定 （GABA）人类癌细胞的水平。 GABA，主要称为抑制性神经递质 哺乳动物中枢神经系统，在外周组织中起作用，以调节细胞增殖，分化， 和迁移。考虑到这些功能在癌症和正常细胞中的重要性，必须 了解SLC45A4在非神经元细胞中GABA代谢的背景下的功能。使用稳定 同位素跟踪，发现SLC45A4不是通过促进GABA吸收而增加细胞GABA水平 通过促进Gaba de从头综合，提出了GABA调节的全新机制 合成。因此，假设SLC45A4编码支持的亚细胞鸟氨酸转运蛋白 GABA生产。该程序将向三个方向前进：AIM 1，确定生化 SLC45A4促进细胞GABA产生的机制； AIM 2，确定亚细胞 SLC45A4转运蛋白和GABA合成的分隔；和目标3，确定基板 SLC45A4的偏好和酶特性。总体目标是确定SLC45A4如何调节DE Novo Gaba综合，有助于人类生物化学的基本知识，并可能提供 针对GABA代谢的新疗法。