Probing the role of AKAPs in breast cancer using stapled peptide inhibitors

使用钉合肽抑制剂探讨 AKAP 在乳腺癌中的作用

• 批准号: 8189598
• 负责人: Eileen J Kennedy
• 金额: $ 18.84万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8189598
• 关键词: A kinase anchoring protein; Affinity; Amino Acids; Binding; Biological; Biological Assay; Biology; Breast; Breast Cancer Cell; Cancer Model; Cancer cell line; Cancerous; Catalytic Domain; Cell Death; Cells; Chemicals; Chemistry; Comparative Study; Complex; Critiques; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Development; Disease; Drug or chemical Tissue Distribution; Elements; Engineering; Environment; Enzymes; Epithelial; Epithelial Cells; Eukaryotic Cell; Event; Exhibits; Foundations; Gene Family; Genetic Polymorphism; Goals; Hereditary Breast Carcinoma; Human Genome; Hydrocarbons; Individual; Intervention; Link; Malignant Neoplasms; Mammary gland; Mass Spectrum Analysis; Measures; Mediating; Membrane Proteins; Methodology; Methods; Methylation; Modeling; Monitor; Peptides; Permeability; Pharmaceutical Preparations; Phenotype; Phosphorylation; Phosphotransferases; Play; Polymorphism Analysis; Positioning Attribute; Property; Protein Isoforms; Protein Kinase; Proteins; Regulation; Research; Resistance; Role; Screening procedure; Set protein; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism; Specificity; Structure; Substrate Specificity; Surface; Therapeutic; Writing; base; cell growth; comparative; computerized data processing; design; improved; inhibitor/antagonist; insight; interest; kinase inhibitor; malignant breast neoplasm; meetings; oncology; physical science; protein expression; protein protein interaction; small molecule; success; tool; tumor

DESCRIPTION (provided by applicant): The protein kinase superfamily is one of the largest protein classes in the human genome. Kinases regulate key signaling processes that have been implicated in development and disease, including cancer. Small molecule kinase modulators have become important therapeutic tools and often target catalytic domains that are among the most structurally and functionally conserved regions of these enzymes. While these tools have provided tremendous insight into kinase activity, specificity has been difficult to achieve due to the high degree of structural conservation among kinases. Further, many protein: protein interactions that regulate kinase activity are mediated through -helical interactions that are difficult to disrupt using small molecule approaches. As a means to circumvent these problems, I propose to develop synthetically modified -helices that will be applied as investigative tools to mediate protein: protein interaction events involved in kinase regulation. As a model for understanding kinase regulation, cAMP-dependent Protein Kinase A (PKA) will be used as a paradigm for probing kinase regulation. PKA has broad substrate specificity, so activity is partly regulated by intracellular localization near specific target subsets via interactions with a diverse set of proteins called A Kinase Anchoring proteins (AKAPs). AKAPs interact directly with the regulatory subunits to localize the tetrameric complex. Single nucleotide polymorphism (SNP) analysis has revealed that single amino acid changes in the PKA Binding Region (AKB) of the AKAP protein D-AKAP2 is linked to a variety of disease states, including familial breast cancer. Synthetically stabilized, isoform-specific peptides will be designed to probe AKAP- mediated interactions with the R-subunits of PKA. Cell signaling events, as monitored by protein phosphorylation and expression changes, will be comparatively analyzed by mass spectrometry to uncover isoform-specific kinase activity in non-cancerous and mammary epithelial cancer cell lines. This will be used as a foundation for studying altered kinase signaling in cancer models. PUBLIC HEALTH RELEVANCE: Greater insight into kinase regulation will allow us to understand the implications of kinases on cancer. By taking a chemical biology approach, we can specifically probe a kinase of interest to study its role in cellular signaling in various cancers and compare this to its role in non-disease states. The written critiques and criteria scores of individual reviewers are provided in essentially unedited form in the "Critique" section below. Please note that these critiques and criteria scores were prepared prior to the meeting and may not have been revised subsequent to any discussions at the review meeting. The "Resume and Summary of Discussion" section above summarizes the final opinions of the committee.

描述(申请人提供)：蛋白激酶超家族是人类基因组中最大的蛋白质类别之一。激酶调节着与发育和疾病(包括癌症)有关的关键信号过程。小分子激酶调节剂已经成为重要的治疗工具，并经常针对这些酶的结构和功能最保守的催化域。虽然这些工具提供了对激酶活性的巨大洞察，但由于激酶之间高度的结构保守，特异性一直难以实现。此外，许多蛋白质：调节激酶活性的蛋白质相互作用是通过螺旋相互作用介导的，而这种相互作用很难使用小分子方法来破坏。为了绕过这些问题，我建议开发人工修饰的螺旋，作为研究工具来调节蛋白质：参与激酶调节的蛋白质相互作用事件。CAMP依赖的蛋白激酶A(PKA)作为理解蛋白激酶调控的模型，将被用来作为探测蛋白激酶调控的范例。PKA具有广泛的底物特异性，因此PKA的活性部分受特定靶标亚群附近的细胞内定位的调节，这是通过与一组称为A激酶锚定蛋白(AKAPs)的不同蛋白质相互作用来实现的。AKAP直接与调节亚基相互作用，定位四聚体复合体。单核苷酸多态性(SNP)分析表明，AKAP蛋白D-AKAP2的PKA结合区(AKB)的单一氨基酸变化与包括家族性乳腺癌在内的多种疾病状态有关。合成稳定的、异构体特异性的多肽将被设计来探索AKAP介导的与PKA R亚基的相互作用。通过蛋白质磷酸化和表达变化监测的细胞信号事件将通过质谱学进行比较分析，以揭示非癌症和乳腺上皮性肿瘤细胞系中异构体特异的激酶活性。这将被用作研究癌症模型中改变的激酶信号的基础。 与公共健康相关：更深入地了解激酶的调节将使我们能够理解激酶对癌症的影响。通过采取化学生物学的方法，我们可以专门探索一种感兴趣的激酶，以研究它在各种癌症中的细胞信号转导中的作用，并将其与非疾病状态下的作用进行比较。 个别评审员的书面评论和标准分数以基本上未经编辑的形式在下面的“评论”部分提供。请注意，这些批评意见和标准分数是在会议之前准备的，在审查会议上进行任何讨论后可能没有加以修订。上文“恢复和总结讨论”一节概述了委员会的最后意见。