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

• 批准号: 9206141
• 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
• 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/9206141
• 关键词: Abraxane; Address; Binding; Biochemistry; Biological Assay; Biology; Biophysics; Breast Cancer Cell; Breast Cancer Model; CD44 gene; Categories; Cell Surface Receptors; Clinical; Complement; Complex; Cyclophilin A; Cyclosporine; Data; Dependence; Disease; Evaluation; Event; Foundations; Genetic; Growth; Growth Factor; Hand; Hormones; Human; Immunologics; Immunosuppression; In Vitro; Injectable; Interleukin-6; Knock-out; Lead; Ligand Binding; Ligands; Mammary Neoplasms; Mediator of activation protein; Metastatic to; Methods; Modeling; Modification; Molecular; Molecular Conformation; Molecular Profiling; Mus; Mutagenesis; NMR Spectroscopy; Neoplasm Metastasis; Pathogenesis; Pathway interactions; Patients; Peptidylprolyl Isomerase; Pharmacology; Phenotype; Phosphotransferases; Population; Positioning Attribute; Process; Prolactin; Prolactin Receptor; Protein Tyrosine Kinase; Proteins; Recombinant Proteins; Regulation; Research; Resources; Role; Signal Transduction; 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; knock-down; loss of function; malignant breast neoplasm; mouse model; overexpression; phase I trial; progenitor; protein protein interaction; public health relevance; receptor; solid state nuclear magnetic resonance; src-Family Kinases; stem

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）：受体相关酪氨酸激酶，例如 Jak2 和 Src，充当配体结合的近端介体。大量数据表明，这两种激酶均对乳腺癌的发病机制有显着影响，但导致其激活的机制仍不确定。我们的研究重点关注乳腺癌相关催乳素受体 (PRLr)，结果表明肽脯氨酰异构酶 (PPI)、亲环蛋白 A (CypA) 是激活这些激酶所必需的。这些发现表明 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 之间多聚体复合物的形成是配体诱导的 PRLr 相关信号复合物激活所必需的。在转化水平上，我们对 CypA 敲除模型的评估以及 PPI 抑制剂环孢素 A (CsA) 和 NIM811 体外和体内乳腺癌模型的成功使用进一步证实了这些结果。总而言之，我们的研究结果使我们假设这些蛋白质与 CypA 的 PPI 活性之间的分子间相互作用导致配体结合后触发 Jak2 和 Src，并且此类事件与乳腺癌的发病机制高度相关。该假设将在三个具体目标中进行测试，如下：首先，将使用乳腺癌细胞内的诱变、过表达和敲低方法来评估 PRLr/Jak2/Src/CypA 内蛋白质-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 光谱将用于评估 PRLr/Jak2/Src 复合物的构象异构体状态，如 CypA 的 PPI 活性所调节。第三，功能获得和功能丧失方法将用于遗传和异种移植小鼠模型，以评估乳腺癌发病过程中 PRLr 复合物内蛋白质相互作用和 PPI 功能的作用。这些研究具有非常重要的意义，因为它们将为我们理解 PPI 活性 CypA 调节的受体 Jak2/Src 激活过程中的结构/功能关系提供分子基础，并将这些发现转化为乳腺癌的细胞模型至小鼠模型。