Regulation of Her2/neu by activation loop phosphorylation and ALG-2

通过激活环磷酸化和 ALG-2 调节 Her2/neu

• 批准号: 7299548
• 负责人: RON BOSE
• 金额: $ 14.32万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-18 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7299548
• 关键词: 1-Phosphatidylinositol 3-Kinase; Affect; Antibodies; Apoptosis; Appendix; Applications Grants; Baculoviruses; Binding; Binding Sites; Biochemical; Biological Assay; Biotinylation; Calcium-Binding Proteins; Cell surface; Cells; Class; Collaborations; Coupled; Cultured Cells; Dimerization; Doctor of Medicine; Doctor of Philosophy; Epidermal Growth Factor Receptor; ErbB Receptor Family Protein; Event; Extracellular Domain; Fellowship; Genes; Goals; Grant; Human; Immunofluorescence Immunologic; In Vitro; Insulin Receptor; Internal Medicine; Knowledge; Laboratories; Length; Link; Lipids; Literature; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Measures; Mediating; Medical; Medical Oncology; Memorial Sloan-Kettering Cancer Center; Mitogen-Activated Protein Kinases; Modeling; Molecular; Monoclonal Antibodies; Mus; Mutate; Mutation; Pathway interactions; Patients; Pattern; Phospholipase; Phosphorylation; Phosphorylation Site; Phosphotransferases; Process; Protein Databases; Protein Kinase; Protein Overexpression; Protein Tyrosine Kinase; Proteins; Proteomics; Publishing; Rate; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Recombinants; Regulation; Research; Research Personnel; Residencies; Resistance; Roche brand of trastuzumab; STAT1 gene; Scientist; Second Messenger Systems; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Site; Small Interfering RNA; Testing; Training Programs; Transgenic Mice; Translational Research; Trastuzumab; Tyrosine; Tyrosine Phosphorylation; Ubiquitination; Universities; Vesicle; Western Blotting; Work; base; cancer therapy; cell transformation; design; dimer; inhibitor/antagonist; kinase inhibitor; knock-down; malignant breast neoplasm; monomer; novel; oncology; outcome forecast; programs; protein protein interaction; receptor; receptor internalization; research study; second messenger; small molecule; src-Family Kinases; ubiquitin ligase

DESCRIPTION (provided by applicant): The candidate, Ron Bose, M.D., Ph.D., is a graduate of the Medical Scientist Training Program held jointly at Cornell University- Rockefeller University- Memorial Sloan-Kettering Cancer Center. His Ph.D. research, conducted at Memorial Sloan Kettering, studied a lipid second messenger pathway which induced apoptosis. Dr. Bose completed residency in Internal Medicine in 2002 and entered Medical Oncology fellowship at Johns Hopkins University. His goals are to pursue basic and translational research by applying proteomics to study signal transduction pathways in breast cancer. He has published a proteomic study of Her2/neu tyrosine kinase signaling (Appendix A) and systematically compared these results to proteomic studies of Epidermal growth factor receptor (EGFR) signaling already in the literature. In this proposal, he plans to pursue two related proteomic findings, Her2/neu activation loop phosphorylation on tyrosine 877 and Her2/neu induced phosphorylation of Apoptosis-linked gene 2 (ALG2), with focused mechanistic experiments. Both activation loop phosphorylation and ALG-2 phosphorylation may be regulatory mechanisms for Her2/neu function and may help control the receptor tyrosine kinase after dimerization has occurred. The activation loop is a major structural feature of kinase domains and has been shown to regulate the activity of insulin receptor kinase and Src. ALG-2 interacts with proteins involved in receptor internalization. Internalization and degradation of Her2/neu is a major mechanism of action of the anti-Her2/neu antibody, trastuzumab (Herceptin). In this proposal, the specific aims will test: 1) if recombinant Her2/neu kinase domain can in vitro autophosphorylate itself on the activation loop and the interaction of this process with a recent model of kinase domain dimerization, 2) if mutation of the activation loop site (Y877F) affects downstream signaling and trastuzumab-induced Her2/neu internalization, 3) if ALG-2 and its interacting protein Alix/AIP1 affect trastuzumab induced Her2/neu internalization and degradation. This research will be performed in the Department of Oncology, Johns Hopkins University. Relevance: Her2/neu and EGFR are protein kinases involved in multiple human cancers. Careful, mechanistic study of their regulation will result in better application of current targeted therapies and possible design of a new class of kinase inhibitors for the treatment of cancer.

描述（由申请人提供）：候选人，罗恩博斯，医学博士，哲学博士、是康奈尔大学-洛克菲勒大学-纪念斯隆-凯特琳癌症中心联合举办的医学科学家培训计划的毕业生。博士学位在Memorial Sloan Kettering进行的一项研究研究了诱导细胞凋亡的脂质第二信使途径。Bose博士于2002年完成了内科住院医师培训，并进入约翰霍普金斯大学的肿瘤内科研究员行列。他的目标是通过应用蛋白质组学研究乳腺癌的信号转导途径来进行基础和转化研究。他发表了Her 2/neu酪氨酸激酶信号传导的蛋白质组学研究（附录A），并将这些结果与文献中已有的表皮生长因子受体（EGFR）信号传导的蛋白质组学研究进行了系统性比较。在这项提案中，他计划进行两个相关的蛋白质组学研究结果，Her 2/neu激活环磷酸化酪氨酸877和Her 2/neu诱导的磷酸化凋亡相关基因2（ALG 2），重点机制实验。激活环磷酸化和ALG-2磷酸化可能是Her 2/neu功能的调节机制，并可能有助于控制二聚化后的受体酪氨酸激酶。激活环是激酶结构域的主要结构特征，并且已显示调节胰岛素受体激酶和Src的活性。ALG-2与参与受体内化的蛋白质相互作用。Her 2/neu的内化和降解是抗Her 2/neu抗体曲妥珠单抗（赫赛汀）的主要作用机制。在本提案中，具体目标将检验：1）如果重组Her 2/neu激酶结构域可以在体外在激活环上自磷酸化自身，以及该过程与激酶结构域二聚化的最新模型的相互作用，2）如果激活环位点（Y877 F）的突变影响下游信号传导和曲妥珠单抗诱导的Her 2/neu内化，3）ALG-2及其相互作用蛋白阿利克斯/AIP 1是否影响曲妥珠单抗诱导的Her 2/neu内化和降解。本研究将在约翰霍普金斯大学肿瘤学系进行。 相关性：Her 2/neu和EGFR是参与多种人类癌症的蛋白激酶。对它们的调节进行仔细的机制研究将导致更好地应用当前的靶向治疗，并可能设计出一类新的激酶抑制剂用于治疗癌症。