Inhibiting Multi-Functional ALDOA for Cancer Therapy

抑制多功能 ALDOA 用于癌症治疗

基本信息

批准号：
10357451
负责人：
GARTH POWIS
金额：
$ 50.46万
依托单位：
PHUSIS THERAPEUTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-12 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10357451
关键词：
1-Phosphatidylinositol 3-Kinase Actins Active Sites Affinity Aldolase A Allosteric Regulation Allosteric Site Binding Binding Proteins Biological Biomass Blood C-terminal Cancer Cell Growth Catalytic Domain Cell Communication Cell Nucleus Cell Proliferation Cell Survival Cell physiology Cells Chemicals Complex Consensus Crystallization Cytoskeletal Proteins Cytoskeleton Cytosol Development Distal E-Cadherin EP300 gene Elements Energy Metabolism Energy Supply Energy-Generating Resources Enzymes Evolution Fibrosis Fructose Fructosediphosphate Aldolase Gene Family Genes Genetic Transcription Glycolysis Glycolysis Inhibition Goals Growth Growth Factor Human Hypoxia Interphase Cell Lead Link Malignant Neoplasms Malignant neoplasm of pancreas Mediating Mesenchymal Molecular Mus Neoplasm Metastasis Nuclear Nuclear Proteins Oncogenes Pancreas Pathway interactions Patients Pharmacology Phenotype Process Prognosis Proteins Reporting Repression Resolution Role Solid Neoplasm Stream Structure Tail Therapeutic Tight Junctions Time Transcription Coactivator Transcriptional Activation Vascular Endothelial Growth Factors X-Ray Crystallography Xenograft Model aerobic glycolysis alpha Tubulin anaerobic glycolysis angiogenesis anti-cancer base cancer cell cancer initiation cancer therapy checkpoint therapy design improved in vivo inhibitor/antagonist member novel novel drug class novel therapeutics pancreatic cancer cells polymerization prevent response small molecule transcription factor tumor tumor growth tumor xenograft

项目摘要

Fructose-bisphosphate aldolase A (ALDOA) is an ancient, highly expressed gene that has acquired three distinct cellular activities. The best understood activity is catalyzing a key step in aerobic glycolysis. ALDOA also has protein binding activities independent of its catalytic activity (“moonlighting”), that include binding to cytoskeletal proteins, which use an “EΦE” motif that fits into a “moonlighting pocket” of ALDO. Binding to the cytoskeleton actin holds ALDO in an inactive form until it is needed, when it is released by an increase in the levels of its substrate fructose-1, 6-bisphosphate and/or by growth-factor-induced PI-3-kinase activity. Another moonlighting function of ALDOA is its presence in the nucleus of cancer cells, where it is associated with increased proliferation. One possibility is that nuclear ALDOA binds to the EΦE motif on EID-1 in the nucleus to inhibit HIF-1's transcriptional co-activator, p300. ALDOA levels are increased in many cancers, particularly pancreatic cancer, where it has been linked to poor patient survival and an increase in metastasis. Pancreatic cancer cell has high levels of anaerobic glycolysis that is further increased by the hypoxia inducible transcription factor-1 (HIF-1). HIF-1 induces ALDOA and other glycolytic enzymes that in turn maintain high HIF-1 activity through an AMPK/p300-dependent feed-forward loop. HIF-1 activity leads to VEGF release and angiogenesis, and the induction of other cancer cell survival mechanisms. Increased glycolysis provides hypoxic cancer cells with an increased supply of energy (ATP) and essential metabolites for biomass synthesis. Elevated ALDOA is also associated with low E-cadherin, a component of cancer cell tight junctions (TJ) necessary for cell-cell interactions, giving a more mesenchymal phenotype and increased tumor metastasis. This most likely is due to binding of ALDOA to the EΦE motif of cytoskeleton actin causing its polymerization and disassembly, or other inactivation of the TJ with loss of E-cadherin. All this makes ALDOA an exceptional druggable anti-cancer target. Our X-ray crystallography studies have identified a new role for the C-terminal domain of ALDO in its catalytic activity, as well as a reactive Cys289 residue that allows allosteric regulation of catalytic activity. We have identified a novel lead probe allosteric inhibitor of ALDOA that forms a complex with Cys289 inhibiting glycolysis, HIF-1 activity and the proliferation of cancer cells, and in vivo inhibits glycolysis and tumor growth in a tumor xenograft model. Using this biologically stable allosteric inhibitor as a lead, one of our goals is to make more potent inhibitors that bind to the allosteric site Cys289, as well as high affinity bifunctional inhibitors that simultaneously engage the active site and moonlighting pocket. We will use X-ray crystallography to help us design inhibitors that modulate nuclear regulators of HIF-1 activity; and not least to design direct inhibitors of ALDOA catalytic activity. We will use these compounds as pharmacological probes to inhibit the various activities of ALDOA, and as potential leads for new therapies for the treatment of pancreatic, and other cancers.

果糖二磷酸醛缩酶 A (ALDOA) 是一种古老的、高度表达的基因，已获得三个最容易理解的活性是催化有氧糖酵解的关键步骤。还具有独立于其催化活性（“兼职”）的蛋白质结合活性，包括与细胞骨架蛋白，它使用“EΦE”基序，适合 ALDO 的“兼职口袋”结合。细胞骨架肌动蛋白使 ALDO 保持非活性状态，直到需要时，通过增加其底物果糖-1, 6-二磷酸水平和/或生长因子诱导的 PI-3-激酶活性。 ALDOA 的兼职功能是它存在于癌细胞的细胞核中，在那里它与一种可能性是核 ALDOA 与细胞核中 EID-1 上的 EΦE 基序结合。为了抑制 HIF-1 的转录共激活因子，p300 水平在许多癌症中升高，尤其是。胰腺癌，它与患者生存率低和转移增加有关。癌细胞具有高水平的无氧糖酵解，缺氧诱导进一步增加无氧糖酵解转录因子-1 (HIF-1) 诱导 ALDOA 和其他糖酵解酶，从而维持高水平。 HIF-1 活性通过 AMPK/p300 依赖性前馈环路导致 VEGF 释放和抑制。血管生成和其他癌细胞存活机制的诱导提供了增加的糖酵解。缺氧的癌细胞，能量 (ATP) 和生物量必需代谢物的供应增加 ALDOA 合成升高还与 E-钙粘蛋白（癌细胞紧密连接的一种成分）水平降低有关。 (TJ) 是细胞间相互作用所必需的，提供更多的间充质表型并增加肿瘤这很可能是由于 ALDOA 与细胞骨架肌动蛋白的 EΦE 基序的结合导致的。聚合和分解，或 TJ 的其他失活以及 E-钙粘蛋白的损失，所有这些都使得 ALDOA 成为可能。我们的 X 射线晶体学研究发现了一种特殊的可药物抗癌靶点。 ALDO 的 C 端结构域的催化活性，以及反应性 Cys289 残基，使得我们已经鉴定出一种新型的 ALDOA 先导探针变构抑制剂。与 Cys289 形成复合物，抑制糖酵解、HIF-1 活性和癌细胞增殖，以及使用这种生物稳定的变构在体内抑制肿瘤异种移植模型中的糖酵解和肿瘤生长。作为先导抑制剂，我们的目标之一是制造更有效的抑制剂，与变构位点 Cys289 结合，如以及同时参与活性位点和兼职口袋的高亲和力双功能抑制剂。我们将使用X射线晶体学来帮助我们设计调节HIF-1活性核调节因子的抑制剂；尤其是设计 ALDOA 催化活性的直接抑制剂，我们将使用这些化合物作为。抑制 ALDOA 各种活性的药理学探针，并作为新疗法的潜在线索胰腺癌和其他癌症的治疗。