Enzyme-Based Platform Technology for Cancers Utilizing PAM: Etiology of Novel Ami

利用 PAM 治疗癌症的酶平台技术：新型 Ami 的病因学

• 批准号: 8794130
• 负责人: Neil R McIntyre
• 金额: $ 10万
• 依托单位: XAVIER UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-03 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8794130
• 关键词: Alcohols; Amides; Androgens; Basal Cell; Biochemistry; Biological; Biological Assay; Biological Markers; C-terminal; Catalysis; Cell Line; Cells; Chemical Structure; Computer Assisted; Computer Simulation; Databases; Development; Disease; Drug Design; Early Diagnosis; Electrostatics; Enzymatic Biochemistry; Enzymes; Etiology; Evaluation; Feasibility Studies; Geometry; Goals; Growth; Growth Factor; Hormones; Immune System Diseases; Impairment; In Vitro; Kinetics; Ligand Binding; Link; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of prostate; Maps; Mechanics; Mediating; Mixed Function Oxygenases; Modeling; Modification; Molecular; Neuroendocrine Cell; Neuropeptides; Neurosecretory Systems; Peptides; Population; Prevention; Production; Prostate; Protocols documentation; Reaction; Recombinants; Relative (related person); Research; Research Personnel; Resources; Rheumatoid Arthritis; Role; Somatotropin; Staging; Stromal Cells; Technology; Testing; Therapeutic; Tissues; Treatment Efficacy; Universities; Up-Regulation; Work; amidation; analog; androgen independent prostate cancer; autocrine; base; deprivation; design; experience; feeding; improved; inhibitor/antagonist; large scale production; malignant breast neoplasm; novel; overexpression; paracrine; peptide hormone; peptidylglycine alpha-amidating monooxygenase; public health relevance; quantum; therapeutic target; tool; tumor; tumor growth

 DESCRIPTION (provided by applicant): Approximately 50% of all mammalian bioactive peptides require a C-terminal -amide moiety for full biological activity. This post-translationa modification requires the sequential enzyme reactions of PHM and PAL domains of bifunctional PAM. Dysregulated peptide amidation by PAM, either overexpression or impairment, is problematic. The upregulation of PAM is observed in a number of cancers, where the amidated peptide hormones serve as autocrine and/or paracrine growth factors facilitating tumor growth. This proposal has selected androgen independent prostate cancer (AI CaP) as a proof-of-concept study to development a platform technology directed towards the rational therapeutic targeting and early detection protocols of cancers that utilize PAM for progression. The contemporary treatment for late stage CaP is an androgen-ablative therapy. Following therapeutic hormone deprivation, cells in the prostate differentiate, increasing the neuroendocrine populations (which contain PAM). Selection towards the androgen-independent (AI) growth of prostate cancer (CaP) is strongly correlated with -amidated neuropeptide hormone production. We contend that PAM/PHM is an excellent therapeutic target as it is the only known enzyme associated with peptide amidation. Our research platform seeks support to develop the experimental tools to map the TS geometry through a combination of KIEs with steady and transient state kinetics. With quantum computational approaches, chemically stable analogues will be selected matching the molecular electrostatic potential (MEP) maps to databases of known compounds. Establishing a roadmap towards therapeutic discovery by combining the approach of mechanistic enzymology and computer aided drug design will be valuable to those studying the numerous PAM-dependent cancers. In addition, our research team will use peptidomic profiling of AI CaP cell lines to isolate and characterize novel -amidated peptide hormones to serve as biomarkers for early detection and improve therapeutic efficacy (vide supra). Subsequent in vitro feeding studies will identify which amidated peptide hormone(s) mediate tumor proliferation. The far reaching goal of identifying potent biomarkers associated with tumor proliferation will be the design of a robust assay for the early diagnosis and prevention of malignant disease.

 描述（由申请人提供）：大约50%的哺乳动物生物活性肽需要C-末端N-酰胺部分才能获得完整的生物活性。这种后修饰需要双功能PAM的PHM和PAL结构域的顺序酶反应。由PAM引起的肽酰胺化失调，无论是过度表达还是受损，都是有问题的。在许多癌症中观察到PAM的上调，其中酰胺化肽激素用作促进肿瘤生长的自分泌和/或旁分泌生长因子。该提案选择雄激素非依赖性前列腺癌（AI CaP）作为概念验证研究，以开发一种平台技术，该技术旨在利用PAM进行进展的合理治疗靶向和癌症早期检测方案。晚期CaP的当代治疗是雄激素消融疗法。在治疗性激素剥夺后，前列腺中的细胞分化，增加神经内分泌群体（其含有PAM）。前列腺癌（CaP）的雄激素非依赖性（AI）生长选择与β-酰胺化神经肽激素的产生密切相关。我们认为PAM/PHM是一个很好的治疗靶点，因为它是唯一已知的与肽酰胺化相关的酶。我们的研究平台寻求支持，以开发实验工具，通过KIE与稳态和瞬态动力学的组合来映射TS几何形状。通过量子计算方法，将选择化学稳定的类似物，将分子静电势（MEP）图与已知化合物的数据库相匹配。通过结合机械酶学和计算机辅助药物设计的方法来建立治疗发现的路线图，对于那些研究众多PAM依赖性癌症的人来说将是有价值的。此外，我们的研究小组将使用AI CaP细胞系的肽组谱分析来分离和表征新的N-酰胺化肽激素，以用作早期检测和改善治疗功效的生物标志物（参见上文）。随后的体外饲养研究将确定哪种酰胺化肽激素介导肿瘤增殖。鉴定与肿瘤增殖相关的有效生物标志物的深远目标将是设计用于恶性疾病的早期诊断和预防的稳健测定。