Metabolic regulation and inhibition of ATP-citrate lyase

ATP-柠檬酸裂解酶的代谢调节和抑制

• 批准号: 10588229
• 负责人: George Burslem
• 金额: $ 62.2万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 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/10588229
• 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; 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; posttranscriptional; tumor; tumor metabolism

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-柠檬酸盐代谢调节的分子机制 裂解酶（ACLY）的活性，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，是生产脂肪酸，胆固醇， 类异戊二烯和蛋白质乙酰化。ACLY活性升高见于代谢性疾病、心血管疾病、 疾病和许多癌症，促进了几种ACLY抑制剂的开发。虽然许多ACLY 已经开发了抑制剂，只有bempedoic酸，其在体内形成活性bempedoyl-CoA加合物。 肝细胞，已经被FDA批准用于治疗用途。肝细胞癌的风险增加 在患有代谢紊乱的个体中，其中许多人可能是用bempedoic酸治疗的候选人; 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K540、K546和K554处的ACLY乙酰化水平升高以及S455处的磷酸化水平升高， S481和编码具有S481的区域的外显子14的保留也与癌症相关，因此也 提示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的多结构域同源四聚体酶，使用柠檬酸盐、CoA和ATP共底物产生 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， 阻碍了对其催化作用的分子机制的理解， 抑制剂的韦伦实验室最近报道了各种与疾病相关的表型， ACLY功能失调; Marmorstein实验室报告了不同年龄组ACLY的冷冻电镜结构， 反应状态，沿着相关的生物化学和生物物理学研究，以阐明 通过ACLY生产乙酰辅酶A。后一项发现引出了几个未解决的问题， ACLY的代谢调节，并为基于结构的更有效和 用于治疗应用的选择性ACLY抑制剂。这些最近的研究现在将韦伦和 Marmorstein实验室共同努力，解决代谢调节知识的重要差距， 通过以下具体目标抑制ACLY：（1）评估代谢结合剂在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，和（3）评估ACLY抑制剂的分子作用模式。所有这些 研究将揭示ACLY活性和调节如何通过结合介导的分子机制， 以及转录后和翻译后修饰，并将导致合理的 开发ACLY药物治疗癌症。