Nicotinic acetylcholine receptors and neuroblastoma

烟碱乙酰胆碱受体与神经母细胞瘤

• 批准号: 8231045
• 负责人: Rae Nishi
• 金额: $ 19.06万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8231045
• 关键词: Abdomen; Accounting; Adolescent; Adrenal Medulla; Age; Aspirate substance; Automobile Driving; Autonomic nervous system; Binding; Biological Assay; Blood - brain barrier anatomy; Bone Growth; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Canada; Cell Line; Cell Proliferation; Cells; Cessation of life; Chest; Child; Data; Disease; Drug Delivery Systems; Future; Genes; Goals; Home environment; Human; Immunodeficient Mouse; In Vitro; Infant; Lentivirus Vector; Liver; Malignant - descriptor; Malignant Neoplasms; Metastatic malignant neoplasm to brain; Mus; Names; Neoplasm Metastasis; Neural Crest; Neuroblastoma; Neurons; Nicotinic Receptors; Patients; Pediatric Hospitals; Pediatric Oncology; Pediatrics; Pharmaceutical Preparations; Phenotype; Preclinical Drug Evaluation; Primary Neoplasm; Proliferating; RNA Interference; Radiation; Regimen; Reporting; Resistance; Site; Skin; Spinal Cord; Staging; Stem cells; Sympathetic Ganglia; Testing; Transcript; Tumor Cell Line; Tumor-Derived; Tumorigenicity; Vermont; Xenograft procedure; base; cancer genomics; cancer therapy; chemotherapy; gene discovery; high throughput screening; in vivo; killings; malignant phenotype; nerve stem cell; neuroblastoma cell; new therapeutic target; novel; novel therapeutics; oncology; outcome forecast; overexpression; precursor cell; progenitor; receptor; self-renewal; tumor; tumorigenic

DESCRIPTION (provided by applicant): Neuroblastoma is a cancer arising from autonomic neuron progenitor cells that occurs in the adrenal medulla or as paraspinal tumors in the abdomen or thorax of infants and young children. If it is identified in children over the age of 1, it is nearly always a highly malignant, aggressive cancer that kills the child within 4-5 years, even after a strenuous chemotherapeutic regimen combined with radiation and a bone marrow transplant. This project will determine whether an unusual nicotinic acetylcholine receptor is a novel target for therapeutics in treating neuroblastoma. We have isolated cells from bone marrow aspirates obtained from patients with stage 4 neuroblastoma. When tested in xenograft assays in immunodeficient mice, the cells can be categorized into tumor-initiating cells (TICs) and non-tumor initiating cells (non-TICs). We have discovered that the gene encoding the a5 nicotinic acetylcholine receptor (nAChR) subunit, CHRNA5, is expressed at 7- 19-fold higher levels in TICs than in non-TICs. Furthermore, CHRNA5 is 2-10-fold higher in TICs than in normal neurogenic neural crest stem cells, and CHRNA5 is elevated in primary neuroblastoma tumors and cell lines derived from tumors (SH-SY5Y, SKN-KCN, SKN-KCNR) above levels found in normal sympathetic ganglia. In a high throughput drug screen for compounds that would reduce proliferation of TICs, MG 624, a nAChR antagonist, was identified. Based upon our expression data, we hypothesize that nicotinic receptors containing a5 contribute to the malignant phenotype of neuroblastoma and therefore may be a new target for drugs because these receptors are uncommon in the normal autonomic nervous system. The proposed aims will test this hypothesis by: 1) using RNAi to determine whether one or more nAChR subunits is necessary for the malignant phenotype of neuroblastoma cells (TICs and cell lines from primary tumors) and their sensitivity to MG 624; 2) determining whether overexpression of CHRNA5 and/or other subunits in normal skin-derived precursors is sufficient to produce a transformed phenotype. PUBLIC HEALTH RELEVANCE: In children under the age of 15, neuroblastoma accounts for 7% of malignancies, yet 15% of pediatric oncology deaths are due to neuroblastoma. Patients with Stage 4 neuroblastoma, which is characterized by widely disseminated tumors, have a particularly poor prognosis, with 25% survival after 4 years, despite stringent chemotherapy combined with radiation and bone marrow transplants. The goal of this project is to determine whether nicotinic receptors are a promising new target for therapeutics in treating aggressive neuroblastoma.

描述（由申请人提供）：神经母细胞瘤是一种由自主神经元祖细胞引起的癌症，发生在肾上腺髓质或婴儿和幼儿腹部或胸部的脊柱旁肿瘤。如果在1岁以上的儿童中发现，它几乎总是一种高度恶性的侵袭性癌症，即使在剧烈的化疗方案结合放射和骨髓移植后，也会在4-5年内杀死儿童。该项目将确定一种不寻常的烟碱乙酰胆碱受体是否是治疗神经母细胞瘤的新靶点。我们从4期神经母细胞瘤患者的骨髓穿刺液中分离出细胞。当在免疫缺陷小鼠中的异种移植物测定中测试时，细胞可被分类为肿瘤起始细胞（TIC）和非肿瘤起始细胞（非TIC）。我们已经发现，编码α 5烟碱乙酰胆碱受体（nAChR）亚基的基因CHRNA 5在TIC中的表达水平比在非TIC中高7- 19倍。此外，TIC中的CHRNA 5比正常神经源性神经嵴干细胞高2-10倍，并且CHRNA 5在原发性神经母细胞瘤肿瘤和源自肿瘤的细胞系（SH-SY 5 Y、SKN-KCN、SKN-KCNR）中升高至高于正常交感神经节中发现的水平。在一个高通量药物筛选的化合物，将减少TIC的增殖，MG 624，nAChR拮抗剂，被确定。基于我们的表达数据，我们假设含有α 5的烟碱受体有助于神经母细胞瘤的恶性表型，因此可能是药物的新靶点，因为这些受体在正常自主神经系统中不常见。提出的目标将通过以下方式来测试这一假设：1）使用RNAi来确定一个或多个nAChR亚基是否是神经母细胞瘤细胞（来自原发性肿瘤的TIC和细胞系）的恶性表型及其对MG 624的敏感性所必需的; 2）确定CHRNA 5和/或正常皮肤来源的前体中的其他亚基的过表达是否足以产生转化的表型。 公共卫生相关性：在15岁以下的儿童中，神经母细胞瘤占恶性肿瘤的7%，但15%的儿科肿瘤死亡是由于神经母细胞瘤。4期神经母细胞瘤患者的特征是肿瘤广泛扩散，预后特别差，4年后的生存率为25%，尽管进行了严格的化疗联合放疗和骨髓移植。该项目的目标是确定烟碱受体是否是治疗侵袭性神经母细胞瘤的有希望的新靶点。