Molecular Therapeutics of Kidney Cancer: MET Gene and BHD Gene

肾癌的分子治疗：MET基因和BHD基因

基本信息

批准号：
7965987
负责人：
William Marston Linehan
金额：
$ 139.43万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7965987
关键词：
Affect American Animal Model Animals Area Binding Cancer Etiology Cancer cell line Cell Surface Receptors Cells Clear Cell Clinical Code Cutaneous Cyst Cystic kidney Cytoplasm Development Disease Evaluation Family Folliculin Foundations Genes Germ-Line Mutation Growth Factor Hepatocyte Growth Factor Hereditary Neoplastic Syndromes Hereditary Papillary Renal Carcinoma Histology Human Hybrids Hyperplasia In Vitro Inherited Kidney Kidney Neoplasms Knockout Mice Lead Loss of Heterozygosity Lung MET Gene Mutation MET gene Modeling Molecular Mus Mutation New Agents Nonsense Mutation Oncogenes Papillary Pathway interactions Patients Phenotype Phosphorylation Protein Binding Proteins Renal Cell Carcinoma Renal carcinoma Risk Role Sampling Sirolimus Somatic Mutation Specimen Syndrome Testing Therapeutic Tissues Tumor Suppressor Genes Tumor Suppressor Proteins Tyrosine Kinase Domain VHL gene Work base cancer type in vitro Model in vivo kindred loss of function meetings novel receptor therapeutic target tumor

Affect American Animal Model Animals Area Binding Cancer Etiology Cancer cell line Cell Surface Receptors Cells Clear Cell Clinical Code Cutaneous Cyst Cystic kidney Cytoplasm Development Disease Evaluation Family Folliculin Foundations Genes Germ-Line Mutation Growth Factor Hepatocyte Growth Factor Hereditary Neoplastic Syndromes Hereditary Papillary Renal Carcinoma Histology Human Hybrids Hyperplasia In Vitro Inherited Kidney Kidney Neoplasms Knockout Mice Lead Loss of Heterozygosity Lung MET Gene Mutation MET gene Modeling Molecular Mus Mutation New Agents Nonsense Mutation Oncogenes Papillary Pathway interactions Patients Phenotype Phosphorylation Protein Binding Proteins Renal Cell Carcinoma Renal carcinoma Risk Role Sampling Sirolimus Somatic Mutation Specimen Syndrome Testing Therapeutic Tissues Tumor Suppressor Genes Tumor Suppressor Proteins Tyrosine Kinase Domain VHL gene Work base cancer type in vitro Model in vivo kindred loss of function meetings novel receptor therapeutic target tumor

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项目摘要

Molecular Therapeutics of Kidney Cancer-MET Gene and BHD Gene Understanding the genes that cause kidney cancer provides the opportunity to develop approaches for molecular therapeutics for this disease. We have identified 3 genes that cause cancer of the kidney: the VHL gene (clear cell renal cell carcinoma); the c-Met gene (papillary type 1 renal carcinoma); and the BHD gene (chromophobe renal carcinoma). Targeting the MET Gene: Type 1 Papillary Kidney Cancer We have found activating mutations of the MET gene in the germline of patients with Hereditary Papillary Renal Cell Carcinoma (HPRC) as well as in a subset of tumors from patients with sporadic, type 1 papillary kidney cnacer Studies are underway to target the c-Met type 1 papillary kidney cancer gene pathway in papillary kidney cancer. The Met gene codes for a cell surface receptor for a systemically circulating growth factor, hepatocyte growth factor (HGF). The germline mutations identified in the HPRC kindreds and somatic mutations of the c-Met oncogene in sporadic type 1 papillary renal carcinoma are located in the tyrosine kinase domain of the MET gene and are predicted to activate this receptor. In-vitro and in-vivo studies are underway to evaluate the role of agents which block this cancer gene pathway as a potential approach for the treatment of type 1 papillary renal carcinoma. Targeting the BHD Gene: Chromophobe Kidney Cancer The BHD gene is the gene for the inherited form of chromophobe kidney cancer associated with Birt-Hogg-Dub syndrome. When we found the BHD gene it was a novel gene with no known function. Studies are currently underway to determine what type cancer gene the BHD gene, how it functions normally and how damage to this gene leads to chromophobe renal carcinoma. We have identified mutations of the BHD gene in 94% of the BHD families tested. In order to determine what type of gene the BHD gene is we searched for mutation of the second copy of the gene in kidney tumor specimens from BHD patients. We found mutation (or loss of heterozygosity) of the second copy (the wild type copy) of the BHD gene in 70% of the tumor samples evaluated. These findings provided the evidence that the BHD gene is a loss of function, tumor suppressor gene. When we found the BHD gene it was a novel gene with unknown function. In order to determine what the function of the BHD gene is we performed studies to determine which proteins bind to the BHD protein (called folliculin). We found that folliculin binds to a novel protein, called FNIP1 (folliculin interacting protein) that FNIP1 binds to AMPK, which is the cells main energy sensing protein. AMPK phosphorylates both FNIP1 and AMPK and FNIP phosphorylate folliculin. AMPK inhibits the function of MTOR through the TS pathway. We found that MTOR phosphorylates folliculin and that this phosphorylation is inhibited in-vitro by treatment with rapamycin. We have subsequently found that folliculin binds to a second protein, FNIP2, which also binds AMPK and which is also phosphorylated by AMPK and that AMPK/FNIP2 phosphorylate folliculin. Folliculin and FNIP1 and FNIP2 co-localize in the cytoplasm and the binding of folliculin to FNIP1 and FNIP2 is in the carboxy terminus of the protein. The finding that the germline BHD mutations are predominantly mutations that are predicted to truncate the protein (frameshift or nonsense mutations) suggests that folliculin binding to FNIP1/FNIP2 is critical to folliculins tumor suppressor function. In order to develop a BHD animal model to further understand the effect of mutation of the BHD gene and to provide a model for evaluation of targeted therapeutics we developed a kidney specific BHD knockout mouse. In this model BHD -/- mice developed large cystic kidneys with areas of hyperplastic tissues. These animals develop renal insufficience and survive for only 30 days. In order to evaluate the effect of a targeted therapeutic approach for the BHD gene pathway the BHD -/- animals were treated with rapamycin. The rapamycin treated animals had a significant diminution in the kidney phenotype and their survival was doubled. We have developed a unique in-vitro model of a human kidney cancer cell line from a BHD patient and are evaluating multiple agents with target the BHD pathway in our in-vivo and in-vitro models. These studies provide the basis for the development of a targeted therapeutic approach for BHD-associated kidney cancer and for a subset of patients with sporadic, non-inherited chromophobe kidney cancer.

肾脏癌-met基因和有限元基因的分子疗法理解引起肾癌的基因为开发该疾病分子疗法的方法提供了机会。我们已经确定了3种引起肾脏癌的基因：VHL基因（透明细胞肾细胞癌）； C-MET基因（乳头状1型肾癌）；和BHD基因（铬虫肾癌）。靶向MET基因：1型乳头状肾癌，我们发现在遗传性乳头状肾细胞癌（HPRC）以及零星的肿瘤中，Met基因的激活突变在零星的肿瘤中，1型1型肾小球肾脏CNACER研究的蛋白酶蛋白酶蛋白酶型蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白酶。 MET基因代码用于系统循环生长因子，肝细胞生长因子（HGF）的细胞表面受体。在HPRC亲戚中鉴定出的种系突变和偶发1型型乳头状肾癌中C-MET癌基因的体细胞突变位于MET基因的酪氨酸激酶结构域中，预计将激活该受体。正在进行的体外研究和体内研究正在进行评估药物的作用，该药物阻止了该癌症基因途径，作为治疗1型乳头状肾癌的潜在方法。靶向BHD基因：铬虫肾癌BHD基因是与Birt-Hogg-Dub综合征相关的遗传形式的遗传形式的基因。当我们发现BHD基因时，它是一个没有已知功能的新基因。目前正在进行研究，以确定BHD基因的癌症基因，其正常功能以及该基因损害如何导致染色体肾脏癌。我们已经确定了94％的BHD家族中BHD基因的突变。为了确定BHD基因是哪种类型的基因，我们在BHD患者的肾脏肿瘤标本中搜索了第二副本的突变。我们发现在评估了70％的肿瘤样品中，BHD基因的第二拷贝（野生型副本）的突变（或杂合性丧失）。这些发现提供了证据表明BHD基因是功能丧失，肿瘤抑制基因。当我们发现BHD基因时，它是一个具有未知功能的新基因。为了确定BHD基因的功能，我们进行了研究，以确定哪些蛋白与BHD蛋白（称为卵泡蛋白）结合。我们发现Folliculin与一种新型蛋白质结合，称为FNIP1（Folliculin相互作用蛋白），该蛋白与AMPK结合，即细胞是主要能量传感蛋白。 AMPK磷酸化FNIP1和AMPK和FNIP磷酸化卵泡蛋白。 AMPK通过TS途径抑制MTOR的功能。我们发现mTOR磷酸化卵泡蛋白，并通过用雷帕霉素处理这种磷酸化被抑制体外。随后，我们发现卵泡蛋白与第二种蛋白FNIP2结合，该蛋白还结合了AMPK，并且也通过AMPK磷酸化，并且AMPK/FNIP2磷酸化卵泡蛋白。 Folliculin和FNIP1和FNIP2在细胞质中共定位，以及卵泡蛋白与FNIP1和FNIP2的结合在蛋白质的羧基末端中。种系BHD突变的发现主要是突变，这些突变被预测会截断蛋白质（移料或废话突变）表明与FNIP1/FNIP2结合的卵泡蛋白对卵泡蛋白抑制肿瘤抑制剂的功能至关重要。为了开发BHD动物模型，以进一步了解BHD基因突变的效果，并提供了评估靶向治疗剂的模型，我们开发了一种肾脏特异性的BHD敲除小鼠。在此型号中，BHD - / - 小鼠开发了具有增生组织区域的大囊性肾脏。这些动物会出现肾脏不足，仅生存30天。为了评估BHD基因途径的靶向治疗方法的影响，用雷帕霉素治疗了BHD - / - 动物。雷帕霉素治疗的动物在肾脏表型中显着降低，其存活率增加了一倍。我们已经开发了来自BHD患者的人肾癌细胞系的独特体外模型，并正在评估我们的体内和维特罗模型中具有BHD途径的多种药物。这些研究为开发了与BHD相关的肾癌和一部分患有零星，非系列的铬虫肾癌的患者的靶向治疗方法提供了基础。