Diagnosis, Pathophysiology And Molecular Biology Of Pheochromocytoma

嗜铬细胞瘤的诊断、病理生理学和分子生物学

• 批准号: 8351166
• 负责人: Karel Pacak
• 金额: $ 163万
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 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/8351166
• 关键词: 3-methoxytyramine; Adolescence; Adrenal Gland Neoplasms; Adrenal Glands; Age; Animal Model; Basic Science; Benign; Biochemical; Biological Markers; Catecholamines; Cells; Characteristics; Chemistry; Childhood; Clinical; Collaborations; DNA; Detection; Development; Diagnosis; Dimensions; Disease; Dopamine; Endocrine; Epinephrine; Etiology; Functional disorder; Gene Mutation; Genes; Genetic; Genetic Markers; Genetic screening method; Genotype; Germ-Line Mutation; Goals; Head and neck structure; Histone Deacetylase Inhibitor; Implant; Incidence; Inherited; Institutes; Interdisciplinary Study; Knowledge; Laboratories; Legal patent; Link; Liquid Chromatography; Location; Luciferases; Malignant Neoplasms; Malignant Pheochromocytoma; Measurement; Measures; Medical; Medical center; Metanephrine; Metastatic Pheochromocytoma; Methods; Modeling; Molecular; Molecular Biology; Monitor; Multiple Endocrine Neoplasia Type 1; Mutation; Necrosis; Neoplasm Metastasis; Neoplasms; Neuroendocrinology; Neurofibromatoses; Neurofibromatosis 1; Neurofibromatosis 2; Norepinephrine; Paraganglioma; Pathogenesis; Pathway interactions; Patients; Pattern; Phenotype; Pheochromocytoma; Plasma; Predisposition; Primary Neoplasm; Production; Radiolabeled; Research Personnel; Retroperitoneal Space; Retrospective Studies; Risk Factors; Secondary to; Sequence Analysis; Stem cells; Succinate Dehydrogenase; Syndrome; Tail; Technology; Therapeutic; Time; Translational Research; United States National Institutes of Health; VHL mutation; Variant; Veins; Von Hippel-Lindau Syndrome; base; cancer therapy; clinical Diagnosis; clinical application; cost effective; disease-causing mutation; driving force; improved; men; molecular marker; novel; patient oriented; patient oriented research; preclinical evaluation; radiotracer; tool; tumor; tumor growth; tumorigenesis

The Section is conducting patient-oriented research about the etiology, pathophysiology, genetics, diagnosis, and treatment of pheochromocytoma (PHEO) and paraganglioma (PGL). Projects include not only translational research-applying basic science knowledge to clinical diagnosis, pathophysiology, and treatment-but also reverse translation research where appreciation of clinical findings leads to new concepts that basic researchers can pursue in the laboratory. In order to achieve our goals, the strategy of the Section is based on the multidisciplinary collaborations among investigators from multiple NIH Institutes and outside medical centers. Our Section links together a patient-oriented component with two bench-level components. The patient-oriented component (Medical Neuroendocrinology) is currently the main driving force for our hypotheses and discoveries. The two bench-level components (Tumor Pathogenesis and Chemistry & Biomarkers) emphasize first, technologies of basic research tailored for pathway and target discovery and second, the development of the discoveries into clinical applications. Clinical aspects of PHEO/PGL PHEOs/PGLs show an earlier age of presentation in patients with hereditary rather than sporadic disease. Whether other factors influence age of disease presentation is unclear. We examined age of disease presentation according to different catecholamine phenotypes and locations of PHEOs/PGLs. Patients with PHEOs/PGLs included 172 with and 183 without an identified germ-line mutation or hereditary syndrome. Differences in plasma concentrations of metanephrine, a metabolite of epinephrine, were used to distinguish epinephrine-producing tumors from those lacking epinephrine production. Patients with epinephrine-producing tumors presented with disease 11 years later (P<0.001) than patients with tumors lacking appreciable epinephrine production. Among patients without evidence of a hereditary condition, those with and without epinephrine-producing tumors had respective meanSE ages of 502 and 422 years (P<0.001) at presentation of disease. Patients with multiple endocrine neoplasia type 2 (MEN2) and neurofibromatosis type 1 (NF1), all with epinephrine-producing tumors, similarly presented with disease at a later age than patients with tumors that lacked appreciable epinephrine production secondary to mutations of the von Hippel-Lindau (VHL) and succinate dehydrogenase (SDHx) genes (402 vs 311 years, P<0.001). Among the latter patients, those with multifocal tumors had a younger age of disease presentation than those with solitary tumors (193 vs 342 years, P<0.001). We concluded that the variations in age of disease presentation associated with different tumor catecholamine phenotypes and locations provide useful information for patient management and suggest origins of PHEOs/PGLs from different chromaffin progenitor cells with variable susceptibility to disease causing mutations Hereditary PHEO/PGL We aimed to investigate the rate of SDHB mutations in patients with metastatic PHEO/PGL whose initial tumor presentation began in childhood or adolescence. From 2000 - 2010, 263 PHEO/PGL patients were evaluated. Of those 263 patients, 125 presented with or were found to have metastatic disease; of these 125 patients, 32 presented with a tumor prior to age 20. Genetic testing for mutations in the VHL, MEN, and SDHB/C/D genes was performed on patients without previously identified genetic mutations. Of the 32 patients who presented with metastatic disease and had their primary tumor in childhood or adolescence, sequence analysis of germline DNA found SDHB mutations in 23 (71.9%), SDHD mutations in 3 (9.4%), VHL mutations in 2 (6.3%) and absence of a known mutation in 4 (12.5%). The majority (78.1%) of these 32 patients presented with primary tumors in an extra-adrenal location (retroperitoneum and head and neck). We concluded that the majority of patients with metastatic PHEO/PGL who presented with a primary tumor in childhood or adolescence had primary extra-adrenal tumors and harbored SDHB mutations. Except for primary tumors located in the head and neck where SDHD genetic testing is advised, we recommended that patients presenting with metastatic PHEO/PGL in childhood or adolescence undergo SDHB genetic testing. Another study assessed whether measurements of plasma metanephrine, normetanephrine and methoxytyramine, the O-methylated metabolites of catecholamines, might help distinguish different hereditary forms of the tumor. Plasma concentrations of O-methylated metabolites were measured by liquid chromatography with electrochemical detection in 173 patients with PHEO/PGL, including 38 with MEN2, 10 with NF1, 66 with VHL syndrome and 59 with mutations of the SDHB or D genes. In contrast to patients with VHL and SDH mutations, all patients with MEN2 and NF1 presented with tumors characterized by increased plasma concentrations of metanephrine (indicating epinephrine production). VHL patients usually showed solitary increases in normetanephrine (indicating norepinephrine production), whereas additional or solitary increases in methoxytyramine (indicating dopamine production) characterized 70% of patients with SDH mutations. Patients with NF1 and MEN2 could be discriminated from those with VHL and SDH mutations in 99% of cases by the combination of normetanephrine and metanephrine. Measurements of plasma methoxytyramine discriminated patients with SDH mutations from those with VHL mutations in a further 78% of cases. The distinct patterns of plasma catecholamine O-methylated metabolites in patients with hereditary PHEO/PGL provide an easily utilized tool to guide cost-effective genotyping of underlying disease-causing mutations. Metastatic PHEO/PGL This retrospective study was focused on clinical, genetic, and histopathologic characteristics of primary metastatic versus primary benign PHEOs. We identified 41 subjects with metastatic PHEO and 108 subjects with apparently benign PHEO. We assessed dimension and biochemical profile of the primary tumor, age at presentation, and time to develop metastases. Subjects with metastatic PHEO presented at a significantly younger age (41.414.7 vs. 50.213.7 years; P<0.001), with larger primary tumors (8.383.27 cm vs. 6.182.75 cm; P<0.001) and secreted norepinephrine more frequently (95.1% vs. 83.3%; P=0.046) compared to subjects with apparently benign PHEOs. No significant differences were found in the incidence of genetic mutations in both groups of subjects (25.7% in the metastatic group and 14.7% in the benign group; P=0.13). From available histopathologic markers of potential malignancy, only necrosis occurred more frequently in subjects with metastatic PHEO (27.6% vs. 0%; P<0.001). The median time to develop metastases was 3.6 years, with the longest interval 24 years. In conclusion, regardless of a genetic background, the size of the primary PHEO and age of first presentation are two independent risk factors associated with the development of metastatic disease An animal model of PHEO: Experimental therapeutic approaches For the rapid advancement and preclinical evaluation of novel therapies for PHEO, it is important to develop reliable and sensitive animal models. To this end, we generated two luciferase models: (1) bioluminescent MTT-luc PHEO cells, expressing luciferase, were injected into the tail vein for experimental metastasis and (2) implanted subcutaneously to monitor tumor growth. U.S. Patent Application No. 61/260,991 was filed on November 13, 2009. The title: methods of diagnosis and treatment of cancer using histone deacetylase inhibitors and radiolabeled metaiodobenzylguanidine.

该科正在以患者为中心开展有关嗜铬细胞瘤（PHEO）和副神经节瘤（PGL）的病因学、病理生理学、遗传学、诊断和治疗的研究。项目不仅包括转化研究——将基础科学知识应用于临床诊断、病理生理学和治疗——还包括逆向转化研究，其中对临床发现的理解导致基础研究人员可以在实验室中追求的新概念。 为了实现我们的目标，该科的战略基于来自多个 NIH 研究所和外部医疗中心的研究人员之间的多学科合作。我们的部分将面向患者的组件与两个工作台级组件连接在一起。以患者为导向的部分（医学神经内分泌学）目前是我们的假设和发现的主要驱动力。两个实验室级组成部分（肿瘤发病机制和化学与生物标志物）首先强调为途径和靶标发现量身定制的基础研究技术，其次强调将发现发展为临床应用。 PHEO/PGL 的临床方面 PHEO/PGL 显示遗传性疾病患者的发病年龄较散发性疾病患者更早。其他因素是否影响发病年龄尚不清楚。我们根据不同的儿茶酚胺表型和 PHEO/PGL 位置检查了疾病表现的年龄。患有 PHEO/PGL 的患者包括 172 名和 183 名未发现已确定的种系突变或遗传综合征的患者。变肾上腺素（肾上腺素的一种代谢物）血浆浓度的差异被用来区分产生肾上腺素的肿瘤和不产生肾上腺素的肿瘤。产生肾上腺素的肿瘤患者比缺乏肾上腺素产生的肿瘤患者晚 11 年发病（P<0.001）。在没有遗传性疾病证据的患者中，患有和不患有产生肾上腺素的肿瘤的患者发病时的平均SE年龄分别为502岁和422岁（P<0.001）。患有 2 型多发性内分泌肿瘤 (MEN2) 和 1 型神经纤维瘤病 (NF1) 的患者均患有产生肾上腺素的肿瘤，同样，与因 von Hippel-Lindau (VHL) 和琥珀酸脱氢酶 (SDHx) 基因突变而缺乏明显肾上腺素产生的肿瘤患者相比，其发病年龄也较晚（402 岁与 311 岁， P<0.001）。在后者中，多灶性肿瘤患者的发病年龄比单发性肿瘤患者更年轻（193 岁 vs 342 岁，P<0.001）。我们得出的结论是，与不同肿瘤儿茶酚胺表型和位置相关的疾病表现年龄的变化为患者管理提供了有用的信息，并表明 PHEO/PGL 起源于不同的嗜铬祖细胞，对引起疾病的突变具有不同的易感性 遗传性 PHEO/PGL 我们的目的是调查初次肿瘤表现开始于儿童期或青春期的转移性 PHEO/PGL 患者的 SDHB 突变率。从 2000 年至 2010 年，对 263 名 PHEO/PGL 患者进行了评估。在这 263 名患者中，125 名出现或被发现患有转移性疾病；在这 125 名患者中，有 32 名在 20 岁之前就患有肿瘤。对先前未发现基因突变的患者进行了 VHL、MEN 和 SDHB/C/D 基因突变的基因检测。在 32 名患有转移性疾病并在儿童或青少年时期患有原发肿瘤的患者中，对种系 DNA 的序列分析发现，23 名患者 (71.9%) 存在 SDHB 突变，3 名患者 (9.4%) 存在 SDHD 突变，2 名患者 (6.3%) 存在 VHL 突变，4 名患者 (12.5%) 不存在已知突变。这 32 名患者中的大多数（78.1%）原发肿瘤位于肾上腺外（腹膜后和头颈部）。我们的结论是，大多数在儿童期或青春期出现原发性肿瘤的转移性 PHEO/PGL 患者患有原发性肾上腺外肿瘤并携带 SDHB 突变。除了建议进行 SDHD 基因检测的头颈部原发性肿瘤外，我们建议儿童或青少年时期出现转移性 PHEO/PGL 的患者接受 SDHB 基因检测。 另一项研究评估了血浆变肾上腺素、去甲变肾上腺素和甲氧基酪胺（儿茶酚胺的 O-甲基化代谢物）的测量是否有助于区分不同的遗传性肿瘤形式。 采用液相色谱法和电化学检测法测量了 173 名 PHEO/PGL 患者的 O-甲基化代谢物血浆浓度，其中包括 38 名 MEN2 患者、10 名 NF1 患者、66 名 VHL 综合征患者和 59 名 SDHB 或 D 基因突变患者。与具有 VHL 和 SDH 突变的患者相比，所有 MEN2 和 NF1 患者均出现肿瘤，其特征是血浆变肾上腺素浓度升高（表明肾上腺素产生）。 VHL 患者通常表现出去甲肾上腺素单独增加（表明去甲肾上腺素产生），而 70% 的 SDH 突变患者则表现为甲氧基酪胺（表明多巴胺产生）额外或单独增加。在 99% 的病例中，通过去甲肾上腺素和变肾上腺素的组合，可以将 NF1 和 MEN2 患者与 VHL 和 SDH 突变患者区分开来。在另外 78% 的病例中，血浆甲氧基酪胺的测量可将 SDH 突变患者与 VHL 突变患者区分开来。遗传性 PHEO/PGL 患者血浆儿茶酚胺 O-甲基化代谢物的独特模式提供了一种易于使用的工具，用于指导潜在致病突变的经济有效的基因分型。 转移性PHEO/PGL 这项回顾性研究的重点是原发性转移性 PHEO 与原发性良性 PHEO 的临床、遗传和组织病理学特征。我们确定了 41 名患有转移性 PHEO 的受试者和 108 名患有明显良性 PHEO 的受试者。我们评估了原发肿瘤的尺寸和生化特征、就诊年龄以及发生转移的时间。与患有转移性 PHEO 的受试者相比，转移性 PHEO 受试者的发病年龄明显更年轻（41.414.7 岁 vs. 50.213.7 岁；P<0.001），原发肿瘤较大（8.383.27 cm vs. 6.182.75 cm；P<0.001），且分泌去甲肾上腺素的频率更高（95.1% vs. 83.3%；P=0.046）。显然是良性的 PHEO。两组受试者基因突变发生率无显着差异（转移组为25.7%，良性组为14.7%；P=0.13）。根据现有的潜在恶性肿瘤的组织病理学标志物，只有坏死在转移性 PHEO 受试者中发生的频率更高（27.6% vs. 0%；P<0.001）。发生转移的中位时间为3.6年，最长间隔为24年。总之，无论遗传背景如何，原发性 PHEO 的大小和首次出现的年龄是与转移性疾病发生相关的两个独立的危险因素 PHEO 动物模型：实验治疗方法 对于 PHEO 新疗法的快速进展和临床前评估，开发可靠且敏感的动物模型非常重要。为此，我们生成了两种荧光素酶模型：（1）将表达荧光素酶的生物发光 MTT-luc PHEO 细胞注射到尾静脉中进行实验转移；（2）皮下植入以监测肿瘤生长。 美国专利申请号61/260,991于2009年11月13日提交。标题：使用组蛋白脱乙酰酶抑制剂和放射性标记的间碘苄基胍诊断和治疗癌症的方法。