NEK2 Over-expression Causes Drug Resistance in Myeloma

NEK2 过度表达导致骨髓瘤耐药

基本信息

批准号：
8099737
负责人：
FENGHUANG ZHAN
金额：
$ 30.09万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8099737
关键词：
Apoptosis Autologous Transplantation Biology Cancer Cell Growth Cancer cell line Cell Cycle Cell Cycle Deregulation Cell Cycle Regulation Cell Death Cell Line Cell Proliferation Cell Survival Cell physiology Cells Cessation of life Characteristics Chemicals Clinical Data Development Diagnosis Disease Disease Progression Disease remission Drug resistance Employee Strikes Enrollment Fibroblasts Gene Expression Profiling Genes Goals Grant Growth High Dose Chemotherapy Immunoprecipitation In Vitro Knowledge Lead Libraries Light MAP Kinase Gene MAPK14 gene MEKs Malignant - descriptor Malignant Neoplasms Mediating Minor Monitor Multiple Myeloma Mus Mutation Patients Pharmaceutical Preparations Phosphotransferases Population Protein-Serine-Threonine Kinases Proteins Protocols documentation Recurrent disease Relapse Research Residual Neoplasm Resistance Role Sampling Screening procedure Signal Pathway Signal Transduction Staging Testing Therapeutic Time Treatment Failure WNT Signaling Pathway Work base cancer cell cell growth cell transformation chemotherapeutic agent chemotherapy cohort design drug development effective therapy experience in vitro activity in vitro testing in vivo in vivo Model inhibitor/antagonist killings kinase inhibitor knock-down malignant phenotype mouse model neoplastic cell novel overexpression preclinical study protein protein interaction public health relevance rapid growth research study response small hairpin RNA small molecule tool translational study tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): The long-term objective of this work is to determine the functional role of NEK2 in promoting cancer progression and to use this knowledge to develop novel therapies. The variability in survival of multiple myeloma patients ranges from only a few months to >10 years. We have previously shown that NEK2- expression is increased in multiple cancers including myeloma; higher levels of NEK2-expression induce cell rapid growth and resistance to multiple chemotherapeutics. This project builds upon our finding that when a specific gene (NEK2) that regulates the cell cycle is overexpressed, patients experience a clinically aggressive form and rapid death of myeloma and other cancers. We have made several striking discoveries supporting the hypothesis that NEK2 over-expression disrupts the normal cycle of tumor cell proliferation, resulting in poor survival for myeloma patients. Our goal to test the hypothesis will be accomplished by execution of three specific aims using both in vitro and in vivo models. Aim1: To examine the role of NEK2 in the development and progression of myeloma. We propose to evaluate whether myeloma cells with high-NEK2 expression are characterized with drug resistance in a large cohort of patients with myeloma at different stages; we will also test the growth and drug-resistance of primary myeloma cells with NEK2 high- or NEK2 low-expression in vitro and in vivo; we will introduce NEK2 into normal fibroblast cells and murine cancer-derived myeloma cell line that have low endogenous NEK2 expression, we will then analyze the effects of this alteration on cell transformation, cellular growth, and response to chemotherapeutic agents. Aim 2: to determine NEK2-mediated signaling pathways in cancer cell proliferation and survival. To achieve this goal, we will determine whether some cell growth related signaling pathways are required for NEK2-mediated cell growth and drug resistance in multiple myeloma. We will examine key substrates that directly interact with the NEK2 protein. Furthermore, gene expression profiling (GEP) will be performed on myeloma samples in remission and at relapse who also have GEP at baseline to identify genes that are regulated by NEK2 at different myeloma stages. Aim 3: to develop novel treatments based on targeting NEK2 or its signaling pathways. Through the screen of kinase inhibitor libraries, we have identified two small molecules that can specifically inhibit NEK2 kinase activity, and induce dramatic cancer cell death in vitro. We will use these molecules as a tool to explore their efficacy in killing myeloma cells in vivo. Inhibitors targeting these signaling pathways will be used alone or in combination with the current used chemotherapeutic drugs to evaluate their antimyeloma activities in vitro and in vivo. PUBLIC HEALTH RELEVANCE: Multiple myeloma is the second most common hematological malignant. There is no effective therapy to cure this cancer. Thus, research is urgently needed to determine how this cancer develop and progress in order to design better therapies. In this grant, we propose studies to identify a novel protein target that may lead to develop better therapies for those aggressive myelomas.

描述（由申请人提供）：这项工作的长期目标是确定NEK2在促进癌症进展中的功能作用，并利用这些知识来开发新的疗法。多发性骨髓瘤患者的存活率差异从仅几个月到10年不等。我们以前已经表明，包括骨髓瘤在内的多种癌症中NEK2-表达增加。较高水平的NEK2表达诱导细胞快速生长和对多种化学疗法的耐药性。该项目基于我们的发现，即当调节细胞周期的特定基因（NEK2）过表达时，患者会出现临床侵略性的形式和骨髓瘤和其他癌症的快速死亡。我们做出了一些引人注目的发现，支持了以下假设：NEK2过表达破坏了肿瘤细胞增殖的正常周期，导致骨髓瘤患者的存活率差。我们测试假设的目标将通过使用体外和体内模型执行三个特定目标来实现。 AIM1：检查NEK2在骨髓瘤发展和进展中的作用。我们建议评估在不同阶段的大量骨髓瘤患者中，具有高NEK2表达的骨髓瘤细胞是否具有耐药性。我们还将用NEK2高或NEK2低表达的体外和体内测试原发性骨髓瘤细胞的生长和药物抗性；我们将将NEK2引入正常的成纤维细胞和鼠类癌衍生的骨髓瘤细胞系，内源性NEK2表达低，然后我们将分析这种改变对细胞转化，细胞生长以及对化学治疗剂的反应的影响。目标2：确定NEK2介导的信号通路癌细胞增殖和存活中。为了实现这一目标，我们将确定多发性骨髓瘤中NEK2介导的细胞生长和耐药性是否需要一些与细胞生长相关的信号通路。我们将检查与NEK2蛋白直接相互作用的关键底物。此外，在缓解和复发时，将对基因样品进行基因表达分析（GEP），后者在基线时也具有GEP，以鉴定在不同骨髓瘤阶段受到NEK2调控的基因。 AIM 3：基于靶向NEK2或其信号通路开发新的治疗方法。通过激酶抑制剂文库的屏幕，我们已经确定了两个可以特异性抑制NEK2激酶活性的小分子，并在体外诱导剧烈的癌细胞死亡。我们将使用这些分子作为探索它们在体内杀死骨髓瘤细胞有效性的工具。靶向这些信号通路的抑制剂将单独使用或与当前使用的化学治疗药物结合使用，以评估其体外和体内的抗乳糖瘤活性。公共卫生相关性：多发性骨髓瘤是第二常见的血液系统恶性肿瘤。没有有效治愈这种癌症的有效疗法。因此，迫切需要进行研究以确定该癌症如何发展和进展，以设计更好的疗法。在这笔赠款中，我们建议研究确定一种新的蛋白质靶标，该蛋白可能会导致为那些侵略性的骨髓瘤开发更好的疗法。