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 - 双磷酸醛糖酶A（Aldoa）是一个古老的高表达基因，已获得三个不同的细胞活性。最好的理解活动是催化有氧糖酵解的关键步骤。阿尔多亚还具有独立于其催化活性（“月光”）的蛋白质结合活性，其中包括与细胞骨架蛋白，它使用“EφE”基序，该基序适合Aldo的“月光袋”。与细胞骨架肌动蛋白以不活跃的形式保持Aldo，直到需要它，当它通过增加而释放其底物果糖-1，6-双磷酸盐和/或通过生长因素诱导的PI-3-激酶活性的水平。其他 Aldoa的月光功能是其在癌细胞核中的存在增殖增加。一种可能性是核aldoa在核1中与EID-1上的EφE基序结合抑制HIF-1的转录共激活因子P300。许多癌症的Aldoa水平有所提高，尤其是胰腺癌与患者生存不良和转移的增加有关。胰癌细胞具有高水平的厌氧性糖酵解，可通过低氧诱导进一步增加转录因子1（HIF-1）。 HIF-1诱导Aldoa和其他糖酵解酶 HIF-1通过AMPK/P300依赖性进料前环的活性。 HIF-1活性导致VEGF释放和血管生成以及其他癌细胞存活机制的诱导。增加糖酵解提供缺氧性癌细胞具有增加的能量供应（ATP）和生物质必需代谢物合成。升高的Aldoa也与低电子钙粘蛋白有关，这是癌细胞紧密连接的组成部分（TJ）细胞 - 细胞相互作用所必需的，提供更多的间充质表型和增加的肿瘤转移。这很可能是由于Aldoa与细胞骨架肌动蛋白的EφE基序结合而引起的聚合和拆卸，或其他E-钙粘着蛋白丧失的TJ。所有这些使阿尔多亚一个杰出的可药物抗癌靶标。我们的X射线晶体学研究已经确定了 Aldo的C末端结构域在其催化活性中以及反应性Cys289居住催化活性的变构调节。我们已经确定了一种新型的铅探针变构抑制剂这形成了Cys289的复合物，可抑制糖酵解，HIF-1活性和癌细胞的增殖以及体内抑制肿瘤特征模型中的糖酵解和肿瘤生长。使用这种生物学稳定的变构抑制剂作为铅，我们的目标之一是使与变构现场CYS289结合的潜在抑制剂增加。以及高亲和力双功能抑制剂，这些抑制剂只需接合活跃的部位和月光袋。我们将使用X射线晶体学来帮助我们设计调节HIF-1活性核调节剂的抑制剂；并非最不重要的是设计直接的Aldoa催化活性抑制剂。我们将使用这些化合物作为药理学问题抑制Aldoa的各种活动，并作为新疗法的潜在潜在客户胰腺和其他癌症的治疗。