Metabolic regulation and inhibition of ATP-citrate lyase

ATP-柠檬酸裂解酶的代谢调节和抑制

基本信息

批准号：
10444160
负责人：
George Burslem
金额：
$ 65.04万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10444160
关键词：
ATP Citrate (pro-S)-Lyase Acetyl Coenzyme A Acetylation Acetyltransferase Acids Binding Binding Sites Biochemical Biological Models Cancer Model Carcinogens Cardiovascular Diseases Catalysis Cell Proliferation Cell model Cells Cholesterol Citrates Coenzyme A Cryoelectron Microscopy Data Deacetylase Dependence Development Diet Disease Enzymes Exclusion Exons FDA approved Fatty Acids Feedback Glucose Goals Hand Hepatocarcinogenesis Hepatocyte Histone Acetylation Human In Vitro Individual Knockout Mice Knowledge Lead Malignant Neoplasms Malignant neoplasm of liver Mediating Metabolic Metabolic Diseases Mitochondria Modification Molecular Molecular Conformation Mus Mutagenesis Normal Cell Oxaloacetates PCAF gene Pharmaceutical Preparations Phenotype Phosphorylation Phosphorylation Site Positioning Attribute Post-Translational Protein Processing Primary carcinoma of the liver cells Production Program Development Protein Acetylation Protein Isoforms Protein Subunits RNA Splicing Reaction Regulation Reporting Risk Role Site Source Structure Testing Therapeutic Uses Validation Work adduct base biophysical analysis cancer cell cancer therapy drug development experimental study genetic regulatory protein in vivo inhibitor inhibitor therapy isoprenoid lipid biosynthesis mouse model mutant novel tumor tumor metabolism

项目摘要

The overall goal of this proposal is to dissect the molecular mechanisms of metabolic regulation of ATP-citrate lyase (ACLY) and to characterize ACLY inhibitors for cancer therapy. ACLY is the predominant source of nucleocytosolic acetyl-CoA, an essential building block for the production of fatty acids, cholesterol, isoprenoids and protein acetylation. Elevated ACLY activity is found in metabolic disorders, cardiovascular diseases and many cancers, prompting the development of several ACLY inhibitors. While many ACLY inhibitors have been developed, only bempedoic acid, which forms an active bempedoyl-CoA adduct in hepatocytes, has been approved by the FDA for therapeutic use. Risk of hepatocellular carcinoma is elevated in individuals with metabolic disorders, many of whom may be candidates for treatment with bempedoic acid; yet metabolic regulation of ACLY activity and its functional role in hepatocellular carcinoma remain poorly understood. Elevated levels of ACLY acetylation at K540, K546 and K554 and phosphorylation at S455 and S481, and retention of exon 14 encoding a region with S481, have also been correlated with cancer, thus also suggesting roles for ACLY posttranslational and posttranscriptional modification in cancer metabolism. ACLY is an ~500 kD multidomain homotetrameric enzyme that uses citrate, CoA and ATP cosubstrates to produce oxaloacetate (OAA) and acetyl-CoA. Until recently, the lack of structural information on intact human ACLY has hampered understanding of its molecular mechanism of catalysis and the structure-based development of inhibitors. The Wellen lab recently reported on various disease-associated phenotypes associated with dysregulated ACLY function; and the Marmorstein lab reported on the cryo-EM structures of ACLY in different reaction states, along with associated biochemical and biophysical studies, to elucidate the molecular basis for acetyl-CoA production by ACLY. The latter findings lead to several unresolved questions underlying the metabolic regulation of ACLY and set the stage for the structure-based development of more potent and selective ACLY inhibitors for therapeutic applications. These recent studies now position the Wellen and Marmorstein labs to work together to resolve important gaps in knowledge in metabolic regulation and inhibition of ACLY, through the following specific aims: (1) Evaluate the role of metabolic binders in ACLY activity, (2) Determine the molecular mechanism of how posttranslational modifications and exon 14 retention impact ACLY regulation, and (3) Evaluate the molecular mode of action of ACLY inhibitors. Together, these studies will reveal the molecular mechanisms for how ACLY activity and regulation is mediated by the binding of metabolites, and posttranscriptional and posttranslational modification and will lead to the rational development of ACLY drugs to treat cancer.

该提案的总体目标是剖析 ATP-柠檬酸代谢调节的分子机制裂解酶 (ACLY) 并表征用于癌症治疗的 ACLY 抑制剂。 ACLY 是主要来源核胞质乙酰辅酶A，是产生脂肪酸、胆固醇、类异戊二烯和蛋白质乙酰化。 ACLY 活性升高见于代谢紊乱、心血管疾病疾病和许多癌症，促使多种 ACLY 抑制剂的开发。虽然许多 ACLY 已经开发出抑制剂，只有 bempedoic 酸，它在体内形成活性 bempedoyl-CoA 加合物肝细胞已被 FDA 批准用于治疗用途。肝细胞癌的风险升高对于患有代谢紊乱的个体，其中许多人可能适合接受 Bempedoic Acid 治疗；然而 ACLY 活性的代谢调节及其在肝细胞癌中的功能作用仍然很差明白了。 K540、K546 和 K554 处的 ACLY 乙酰化水平以及 S455 和 K555 处的磷酸化水平升高 S481 以及编码 S481 区域的外显子 14 的保留也与癌症相关，因此表明 ACLY 翻译后和转录后修饰在癌症代谢中的作用。 ACLY 是一种约 500 kD 的多域同四聚酶，使用柠檬酸盐、CoA 和 ATP 共底物来产生草酰乙酸 (OAA) 和乙酰辅酶 A。直到最近，由于缺乏完整的人类 ACLY 的结构信息，阻碍了对其催化分子机制的理解以及基于结构的开发抑制剂。 Wellen 实验室最近报告了多种与疾病相关的表型 ACLY 功能失调； Marmorstein 实验室报告了 ACLY 在不同环境中的冷冻电镜结构反应状态以及相关的生物化学和生物物理研究，以阐明其分子基础 ACLY 生产乙酰辅酶A。后者的发现导致了几个未解决的问题 ACLY 的代谢调节，为基于结构的更有效和更有效的开发奠定了基础用于治疗应用的选择性 ACLY 抑制剂。这些最近的研究现在将 Wellen 和 Marmorstein 实验室将共同努力解决代谢调节和通过以下具体目标抑制 ACLY：（1）评估代谢结合物在 ACLY 中的作用 (2) 确定翻译后修饰和外显子 14 保留的分子机制影响 ACLY 调节，以及 (3) 评估 ACLY 抑制剂的分子作用模式。在一起，这些研究将揭示 ACLY 活性和调节如何通过结合介导的分子机制代谢物、转录后和翻译后修饰，并将导致合理的开发治疗癌症的 ACLY 药物。