Bi-directional regulation of chemokine receptor signaling

趋化因子受体信号传导的双向调节

• 批准号: 10317369
• 负责人: Adriano Marchese
• 金额: $ 31万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10317369
• 关键词: A kinase anchoring protein; Address; Anatomy; BLR1 gene; Binding Proteins; Biochemical; CXCL12 gene; CXCL13 gene; CXCR4 Receptors; Cancer Etiology; Cancer cell line; Cell membrane; Cell physiology; Cells; Cervical; Cessation of life; Clinical; Data; Distant; Family; G-Protein-Coupled Receptors; Growth; Health; Hela Cells; Heterotrimeric GTP-Binding Proteins; In Vitro; Knowledge; Ligands; Ligation; Link; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mediating; Neoplasm Metastasis; Phorbol Esters; Physiological; Play; Prognosis; Protein Family; Protein Isoforms; Protein Kinase C; Proteins; Publishing; Receptor Signaling; Regulation; Reporting; Role; Scaffolding Protein; Serine; Signal Pathway; Signal Transduction; Small Interfering RNA; Specific qualifier value; Tail; Testing; Time; base; biophysical techniques; cancer cell; cell motility; chemokine receptor; desensitization; in vivo; insight; lymph nodes; new therapeutic target; novel; protein activation; protein complex; receptor; response; therapeutic development; trafficking; tumor growth; tumor progression

PROJECT SUMMARY Cervical cancer remains one of the leading causes of cancer-related deaths world-wide. A large body of evidence indicates that a sub-family of G protein-coupled receptors (GPCRs), known as chemokine receptors, are linked to progression of several cancers, including cervical cancer. Dysregulated signaling of certain chemokine receptors correlates with poor prognosis in cervical cancer, yet the mechanisms remain poorly understood. The overall objective of this proposal is to determine the mechanisms governing chemokine receptor regulation in the context of cervical cancer growth and metastasis. This is significant because these studies will define new targets for clinical potential. Specifically, we will focus on a novel paradigm that governs chemokine receptor bi-directional regulation by A kinase anchoring proteins (AKAPs). AKAPs are scaffolding proteins that bind and nucleate multiple proteins, usually belonging to a common signaling pathway, to spatially and temporally control signaling to drive physiological responses. We performed a siRNA screen of AKAPs expressed in HeLa cells, a cervical cancer cell line, and provide evidence that AKAPs are involved in cross- regulation of chemokine receptor trafficking. Further, using biochemical and biophysical approaches we provide evidence that chemokine receptors reside in a compartment with AKAPs and protein kinas C (PKC), but that other GPCRs are excluded. It is possible that chemokine receptors might co-reside in a sub- compartment at the plasma membrane that enables their cross-regulation without input from other GPCRs, likely to exclusively fine tune their signaling. Based on published and new preliminary data we hypothesize that chemokine receptors are part of a multimeric protein complex compartmentalized by AKAPs that governs their bi-directional regulation. To test this hypothesis we will pursue three specific aims. Aim 1: To determine the role of PKC in bi-directional chemokine receptor regulation; Aim 2: To determine the role of AKAPs in chemokine receptor bi-directional trafficking and signaling; and Aim 3: To determine the role of bi-directional chemokine receptor regulation in cervical cancer in vitro and in vivo. At the conclusion of the proposed studies we expect to determine the mechanism of bi-directional regulation of chemokine receptor signaling. Importantly, we expect to establish that novel aspects of chemokine receptor regulation and signaling could be targeted to treat cervical cancer.

项目总结 宫颈癌仍然是全球癌症相关死亡的主要原因之一。一大堆 有证据表明，G蛋白偶联受体(GPCRs)的一个亚家族，即趋化因子受体， 与几种癌症的进展有关，包括宫颈癌。某些基因异常的信号传导 趋化因子受体与宫颈癌预后不良有关，但其机制尚不清楚。 明白了。这项提案的总体目标是确定控制趋化因子的机制 宫颈癌生长和转移过程中的受体调控。这一点意义重大，因为 研究将为临床潜力确定新的目标。具体地说，我们将重点介绍一种新的范式，它管理 A激酶锚定蛋白对趋化因子受体的双向调节作用AKAP正在搭建脚手架 结合并使多个蛋白质成核的蛋白质，通常属于一个共同的信号通路，在空间上 并在时间上控制信号以驱动生理反应。我们对AKAP进行了siRNA筛选 在宫颈癌细胞系HeLa细胞中表达，为AKAP参与交叉调控提供证据。 趋化因子受体运输的调控。此外，使用生化和生物物理方法，我们 提供证据表明趋化因子受体与AKAP和蛋白激动素C(PKC)位于同一隔间， 但其他GPCR被排除在外。趋化因子受体可能共同存在于亚细胞内。 质膜上的隔室，使它们能够在不需要其他GPCR输入的情况下进行交叉调节， 很可能会专门微调它们的信号。根据已公布的和新的初步数据，我们假设 趋化因子受体是由AKAP分隔的多聚体蛋白复合体的一部分，它支配着它们的 双向调控。为了验证这一假设，我们将追求三个具体目标。目标1：确定 PKC在双向趋化因子受体调节中的作用；目的2：确定AKAPs在 趋化因子受体双向运输和信号传递；和目标3：确定双向作用 趋化因子受体在体内外对宫颈癌的调节作用。在拟议的研究结束时 我们希望确定趋化因子受体信号的双向调节机制。 重要的是，我们希望确定趋化因子受体调节和信号的新方面可能是 有针对性地治疗宫颈癌。