CAMKV Kinase Signaling in Neuroblastoma

神经母细胞瘤中的 CAMKV 激酶信号转导

• 批准号: 10752785
• 负责人: JIANHUA YANG
• 金额: $ 44.68万
• 依托单位: CHILDREN'S RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752785
• 关键词: CAMKV; Kinase; Signaling; Neuroblastoma

Neuroblastoma (NB) is the most common extracranial malignant solid tumor in children. High-risk NB remains an incurable disease. Since recurrent somatic mutations in NB occur with relative paucity, dysregulation of oncogenic signal transduction may contribute to NB tumorigenesis. This proposal will pursue that goal by defining the role of CaM kinase-like vesicle-associated (CAMKV) in NB development and examining the therapeutic potential of CAMKV kinase inhibition for treating NB in mouse models. In our preliminary studies, we have found that CAMKV, structurally a member of the calcium/calmodulin-dependent protein kinase family, is quite specifically overexpressed in NB tumor samples, and its high expression predicts poor patient outcome. Furthermore, CAMKV acts as an active CREB kinase in NB cells. Knockdown of CAMKV expression caused a cell proliferation defect and decreased phospho-CREB level in the NB cell lines tested. In addition, we identified K252a and OTSSP167 as CAMKV inhibitors that potently inhibited cell proliferation of several NB cell lines in culture and tumor growth in the xenograft mouse model. Interestingly, our recently generated Camkv knockout mice are viable and fertile. Our long term goal is to develop novel targeted therapies for children with high-risk NB. The objective of this application is to understand the role of CAMKV in NB tumor development and demonstrate the utility of CAMKV as a biomarker and kinase target for NB therapy.The central hypothesis of this work is that CAMKV plays an important role in NB development and is a molecular kinase target in NB. The proposed experiments will test this hypothesis by dissecting the function of CAMKV signaling in NB cells and determining the effect of CAMKV inhibition on NB tumor growth in xenograft mouse models. We will also use recently generated Camkv knockout mice to define the role of CAMKV in NB development in the Th-MYCN transgenic mouse model. The specific aims for this project are: 1) To determine the mechanism of CAMKV regulation and function in NB cells; 2) To determine the role of CAMKV in NB development in tumor mouse models; 3) To determine the therapeutic potential of CAMKV inhibition in vivo. This contribution is significant because it will demonstrate the critical role of CAMKV kinase in NB development and prove that CAMKV kinase inhibition is a potential strategy to treat NB and overcome chemotherapy resistance in vivo. Furthermore, uncovering the regulation of CAMKV kinase signaling will generate a blueprint for CAMKV-based design of treatment for refractory high-risk NB. The proposed research is innovative since it is the first study to specifically uncover and target the CAMKV signaling for designing NB therapeutics and to specifically study CAMKV inhibition by small molecule inhibitors as a mechanism for overcoming chemotherapy resistance. This project will firmly establish CAMKV- based therapy as a novel therapeutic strategy for refractory high-risk NB.

神经母细胞瘤（NB）是儿童颅外最常见的恶性实体瘤。高危NB 仍然是不治之症。由于NB中的复发性体细胞突变相对较少发生， 致癌信号转导的失调可能有助于NB肿瘤的发生。该提案将继续 通过定义CaM激酶样囊泡相关（CAMKV）在NB发展中的作用， 检查CAMKV激酶抑制用于治疗小鼠模型中的NB的治疗潜力。在我们 初步研究，我们发现CAMKV，结构上的钙/钙调素依赖的成员， 蛋白激酶家族，在NB肿瘤样品中特异性过表达，其高表达预示着 患者预后差。此外，CAMKV在NB细胞中充当活性CREB激酶。敲低 CAMKV表达导致NB细胞增殖缺陷和磷酸化CREB水平降低 测试.此外，我们鉴定了K252 a和OTSSP 167作为CAMKV抑制剂，其有效地抑制细胞凋亡。 培养物中几种NB细胞系的增殖和异种移植小鼠模型中的肿瘤生长。有趣的是， 我们最近生产的Camerin基因敲除小鼠可以存活并繁殖。 我们的长期目标是为高危NB儿童开发新的靶向治疗。的目标 本申请旨在了解CAMKV在NB肿瘤发展中的作用，并证明CAMKV的实用性。 CAMKV作为NB治疗的生物标志物和激酶靶点。 在NB的发展中起重要作用，是NB中的分子激酶靶点。拟议的实验 将通过解剖NB细胞中CAMKV信号传导的功能并确定 CAMKV对异种移植小鼠模型中NB肿瘤生长的抑制。我们还将使用最近生成的Cameroon 在Th-MYCN转基因小鼠模型中，使用CAMKV基因敲除小鼠来确定CAMKV在NB发育中的作用。的 本课题的具体目标是：1）明确CAMKV在NB中的调控和作用机制 2）确定CAMKV在肿瘤小鼠模型中NB发展中的作用; 3）确定CAMKV在肿瘤小鼠模型中的作用。 CAMKV抑制在体内的治疗潜力。这一贡献意义重大，因为它将证明 CAMKV激酶在NB发展中的关键作用，并证明CAMKV激酶抑制是一种潜在的 治疗NB和克服体内化疗耐药性的策略。此外，揭露监管 CAMKV激酶信号传导将为基于CAMKV的难治性高风险患者治疗设计蓝图 NB.拟议的研究是创新的，因为它是第一项专门揭示和针对 用于设计NB治疗剂的CAMKV信号传导和特异性研究小分子药物对CAMKV的抑制 抑制剂作为克服化疗耐药性的机制。该项目将坚定地建立CAMKV- 基础疗法作为难治性高危NB的新型治疗策略。