Prolyl isomerase function during Jak Stat signaling in breast cancer

脯氨酰异构酶在乳腺癌 Jak Stat 信号传导过程中的功能

基本信息

批准号：
8814187
负责人：
Charles V Clevenger
金额：
$ 30.97万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8814187
关键词：
A Mouse Abraxane Address Binding Biochemistry Biological Assay Biology Biophysics Boxing Breast Cancer Cell Breast Cancer Model Breeding CD44 gene Categories Cell Surface Receptors Complement Complex Cyclophilin A Cyclosporine Data Dependence Disease Evaluation Event Foundations Genetic Growth Growth Factor Hand Health Hormones Human Immunosuppression In Vitro Interleukin-6 Knock-out Lead Ligand Binding Ligands Mammary Neoplasms Mediator of activation protein Metastatic to Methods Modeling Modification Molecular Molecular Profiling Mus Mutagenesis NMR Spectroscopy Neoplasm Metastasis Pathogenesis Pathway interactions Patients Peptidylprolyl Isomerase Phenotype Phosphotransferases Population Positioning Attribute Process Prolactin Receptor Protein Tyrosine Kinase Proteins Receptor Signaling Recombinant Proteins Regulation Research Resources Role Signal Transduction Solutions Somatotropin Structure Structure-Activity Relationship Testing Therapeutic Toxic effect Transgenic Organisms Translating Tyrosine Universities Ursidae Family Xenograft Model Xenograft procedure analog base conformer design improved in vivo inhibitor/antagonist innovation insight intermolecular interaction loss of function malignant breast neoplasm mouse model overexpression phase I trial progenitor protein protein interaction receptor solid state nuclear magnetic resonance src-Family Kinases stem

项目摘要

DESCRIPTION (provided by applicant): Receptor-associated tyrosine kinases, such as Jak2 and Src, serve as proximal mediators of ligand binding. Abundant data indicate that both of these kinases significantly contribute to the pathogenesis of breast cancer, and yet the mechanisms leading to their activation has remained uncertain. Our research, which has focused on the breast cancer-relevant receptor for prolactin (PRLr), has revealed that the peptidly prolyl isomerase (PPI), cyclophilin A (CypA) is required for the activation of these kinases. These findings would indicate that PPI activity of CypA is involved in a conformational restructuring of this receptor-kinase complex that contributes to its activation. Additional studie have revealed that the formation of a multimeric complex between the PRLr, Jak2, Src, and CypA is necessary of ligand-induced activation of the PRLr-associated signaling complex. At a translational level, these results have been further corroborated by our evaluation of a CypA knockout model and the successful use in breast cancer models of the PPI inhibitors cyclosporine A (CsA) and NIM811 both in vitro and in vivo. Taken together, our findings lead us to hypothesize that the intermolecular interactions between these proteins and the PPI activity of CypA result in the triggering of Jak2 and Src following ligand engagement, and that such events are highly relevant to the pathogenesis of breast cancer. This hypothesis will be tested in three specific aims, as follows: First, the functional role of protein- protein interactions withinthe PRLr/Jak2/Src/CypA will be evaluated using mutagenic, overexpression, and knockdown approaches within breast cancer cells. Second, in vitro kinase and soluble and solid state NMR spectroscopy will be used to assess conformer status with the PRLr/Jak2/Src complex as regulated by the PPI activity of CypA. Third, both gain- and loss-of-function approaches will be used in genetic and xenograft-based murine models to assess the role of protein interactions and PPI function within the PRLr complex during the pathogenesis of mammary cancer. The studies proposes are highly significant in that the will provide a molecular foundation for our understanding of the structure/function relationships during receptor-Jak2/Src activation as modulated by the PPI activity CypA and translate these discoveries through cellular to mouse models of breast cancer.

描述（由申请人提供）：与受体相关的酪氨酸激酶，例如JAK2和SRC，是配体结合的近端介体。大量数据表明，这两种激酶都显着有助于乳腺癌的发病机理，但导致其激活的机制仍然不确定。我们的研究集中在催乳素（PRLR）的乳腺癌相关受体上，揭示了肽蛋白蛋白蛋白蛋白酶（PPI），细胞周期蛋白A（CYPA）激活这些激酶所必需的。这些发现将表明CYPA的PPI活性参与了该受体激酶复合物的构象重组，从而有助于其激活。其他研究表明，PRLR，JAK2，SRC和CYPA之间的多聚体复合物是配体诱导的PRLR相关信号复合物的激活所必需的。在翻译水平上，通过评估CYPA基因敲除模型以及在PPI抑制剂Cyclosporine A（CSA）和NIM811的乳腺癌模型中的成功使用，进一步证实了这些结果。综上所述，我们的发现使我们假设这些蛋白质之间的分子间相互作用与CYPA的PPI活性导致配体参与后的JAK2和SRC触发，并且此类事件与乳腺癌的发病机理高度相关。该假设将以三个特定目的进行检验，如下：首先，将使用诱变，过表达和乳腺癌细胞中蛋白质蛋白质相互作用的功能作用。其次，体外激酶以及可溶性和固态NMR光谱法将用于评估由CYPA的PPI活性调节的PRLR/JAK2/SRC复合物的构象体状态。第三，基于遗传和异种移植的鼠模型将使用功能丧失方法来评估乳腺癌发病机理中PRLR复合物在PRLR复合物中蛋白质相互作用和PPI功能的作用。该研究提出的非常重要的是，该研究将为我们理解由PPI活性CYPA调节的受体-JAK2/SRC激活期间的结构/功能关系的分子基础，并通过细胞将这些发现转化为乳腺癌的小鼠模型。