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.

项目摘要 宫颈癌仍然是全球与癌症相关死亡的主要原因之一。一大堆 证据表明，G蛋白偶联受体（GPCR）的亚家族，称为趋化因子受体， 与包括宫颈癌在内的几种癌症的进展有关。某些信号失调 趋化因子受体与宫颈癌预后不良相关，但机制仍然很差 理解。该提案的总体目的是确定控制趋化因子的机制 受体调节在宫颈癌生长和转移的背景下。这很重要，因为这些 研究将定义临床潜力的新目标。具体而言，我们将重点关注一个政府的新颖范式 趋化因子受体双向调节锚定蛋白（AKAP）。 Akap是脚手架 结合并成核多种蛋白质的蛋白质，通常属于公共信号通路，与空间上 并在时间上控制信号传导以驱动生理反应。我们进行了Akap的siRNA屏幕 在HeLa细胞（一种宫颈癌细胞系）中表达，并提供了AKAP参与交叉的证据 调节趋化因子受体运输。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居住在Akap和蛋白Kinas C（PKC）的隔室中， 但是其他GPCR被排除在外。趋化因子受体可能会在亚趋 质膜处的隔室，可以使其交叉调节，而无需其他GPCR的输入， 可能只能微调其信号传导。根据已发布和新的初步数据，我们假设 趋化因子受体是由AKAP划分的多聚体蛋白质复合物的一部分 双向调节。为了检验这一假设，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；目标2：确定AKAP在 趋化因子受体双向运输和信号传导；目标3：确定双向的作用 体外和体内宫颈癌的趋化因子受体调节。在拟议的研究结束时 我们期望确定趋化因子受体信号传导的双向调节机制。 重要的是，我们期望确定趋化因子受体调节和信号传导的新方面 针对治疗宫颈癌的目标。