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: 10471999
• 负责人: Adriano Marchese
• 金额: $ 30.61万
• 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/10471999
• 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.

项目概要 宫颈癌仍然是全世界癌症相关死亡的主要原因之一。庞大的身躯 有证据表明，G 蛋白偶联受体 (GPCR) 的一个亚家族（称为趋化因子受体） 与包括宫颈癌在内的多种癌症的进展有关。某些信号传导失调 趋化因子受体与宫颈癌的不良预后相关，但其机制仍不清楚 明白了。该提案的总体目标是确定趋化因子的控制机制 宫颈癌生长和转移背景下的受体调节。这很重要，因为这些 研究将为临床潜力确定新的目标。具体来说，我们将重点关注一种新的范式来管理 趋化因子受体通过 A 激酶锚定蛋白 (AKAP) 进行双向调节。 AKAP 是脚手架 结合并成核多种蛋白质的蛋白质，通常属于共同的信号传导途径，在空间上 并暂时控制信号以驱动生理反应。我们对 AKAP 进行了 siRNA 筛选 在 HeLa 细胞（一种宫颈癌细胞系）中表达，并提供证据表明 AKAP 参与交叉 趋化因子受体运输的调节。此外，利用生物化学和生物物理方法，我们 提供证据表明趋化因子受体位于具有 AKAP 和蛋白激酶 C (PKC) 的隔室中， 但其他 GPCR 被排除在外。趋化因子受体可能共存于亚 质膜上的隔室，无需其他 GPCR 的输入即可实现交叉调节， 可能专门微调他们的信号。根据已发布的和新的初步数据，我们假设 趋化因子受体是由 AKAP 分隔的多聚蛋白复合物的一部分，AKAP 控制其 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 宫颈癌的体外和体内趋化因子受体调节。拟议研究的结论 我们期望确定趋化因子受体信号传导双向调节的机制。 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可能是 旨在治疗宫颈癌。