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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：
10926117
负责人：
William Marston Linehan
金额：
$ 95.39万
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/10926117
关键词：
Affect American Animal Model Animals Area Binding Binding Proteins Cancer Etiology Cancer cell line Cell Surface Receptors Cells Clear Cell Clear cell renal cell carcinoma Clinical Code Cutaneous Cyst Cystic kidney Cytoplasm Development Disease Evaluation Family Folliculin Foundations Frameshift Mutation Genes Germ-Line Mutation Growth Factor HGF gene Hereditary Neoplastic Syndromes Hereditary Papillary Renal Carcinoma Histology Human Hybrids Hyperplasia In Vitro Inherited Kidney Kidney Neoplasms Knockout Mice Loss of Heterozygosity Lung MET Gene Mutation MET Oncogene MET gene Modeling Molecular Mus Mutation New Agents Nonsense Mutation Oncogenes Oxyphil Cells Papillary Pathway interactions Patients Phenotype Phosphorylation Protein Truncation 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 cancer type in vitro Model in vivo kindred loss of function novel patient subsets receptor targeted agent targeted treatment tumor

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.

肾癌的分子治疗-MET基因和BHD基因：了解导致肾癌的基因提供了发展这种疾病的分子治疗方法的机会。我们已经确定了3种导致肾癌的基因：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（嫌色细胞肾癌）。靶向MET基因-1型乳头状肾癌：我们已经在遗传性乳头状肾细胞癌（HPRC）患者的生殖系中以及来自散发性1型乳头状肾癌患者的肿瘤子集中发现MET基因的激活突变。靶向乳头状肾癌中的c-Met 1型乳头状肾癌基因通路的研究正在进行中。Met基因编码全身循环生长因子肝细胞生长因子（HGF）的细胞表面受体。在散发性1型乳头状肾癌中发现的HPRC激酶和c-Met癌基因体细胞突变的种系突变位于MET基因的酪氨酸激酶结构域，并被预测激活该受体。体外和体内研究正在进行中，以评估阻断这种癌症基因通路的药物作为治疗1型乳头状肾癌的潜在方法的作用。靶向BHD基因-嫌色肾癌：BHD基因是与Birt-Hogg-Dub综合征相关的遗传性嫌色肾癌的基因。当我们发现BHD基因时，它是一个未知功能的新基因。目前正在进行研究，以确定BHD基因是哪种类型的癌症基因，它如何正常发挥作用，以及该基因的损伤如何导致嫌色肾癌。我们在94%的BHD家族中发现了BHD基因突变。为了确定BHD基因是什么类型的基因，我们在BHD患者的肾脏肿瘤标本中寻找该基因的第二拷贝的突变。我们在70%的肿瘤样本中发现了BHD基因的第二个拷贝（野生型拷贝）的突变（或杂合性丢失）。这些结果为BHD基因是一个功能缺失的抑癌基因提供了证据。当我们发现BHD基因时，它是一个功能未知的新基因。为了确定BHD基因的功能是什么，我们进行了研究，以确定哪些蛋白质与BHD蛋白（称为卵泡素）结合。我们发现卵泡素与一种新的蛋白质FNIP 1（folliculin interacting protein）结合，FNIP 1与细胞主要的能量敏感蛋白AMPK结合。AMPK磷酸化FNIP 1和AMPK，FNIP磷酸化卵泡素。AMPK通过TS通路抑制MTOR的功能。我们发现MTOR使卵泡素磷酸化，并且这种磷酸化在体外通过用雷帕霉素处理而被抑制。我们随后发现，卵泡素与第二种蛋白质FNIP 2结合，FNIP 2也与AMPK结合，也被AMPK磷酸化，AMPK/FNIP 2磷酸化卵泡素。滤泡蛋白与FNIP 1和FNIP 2共定位于细胞质中，并且滤泡蛋白与FNIP 1和FNIP 2的结合在蛋白质的羧基末端。生殖系BHD突变主要是预测截短蛋白质的突变（移码或无义突变），这一发现表明卵泡素与FNIP 1/FNIP 2的结合对卵泡素肿瘤抑制功能至关重要。为了开发BHD动物模型以进一步了解BHD基因突变的影响并提供用于评估靶向治疗的模型，我们开发了肾特异性BHD敲除小鼠。在该模型中，BHD -/-小鼠产生了大的囊性肾，伴有增生组织区域。这些动物发展为肾损害，仅存活30天。为了评估BHD基因途径的靶向治疗方法的效果，用雷帕霉素处理BHD -/-动物。雷帕霉素处理的动物的肾脏表型显著减少，并且它们的存活率加倍。我们已经开发了一种独特的BHD患者人肾癌细胞系体外模型，并正在评估体内和体外模型中靶向BHD途径的多种药物。这些研究为BHD相关肾癌和散发性非遗传性嫌色细胞肾癌患者亚组的靶向治疗方法的开发提供了基础。