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: 7594209
• 负责人: Karel Pacak
• 金额: $ 44.35万
• 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/7594209
• 关键词: Abdomen; Adrenal Glands; Aggressive behavior; Animal Model; Animals; Area; Basic Science; Behavior; Biochemical; Biological Markers; Blood Pressure; Body Regions; CGB gene; CHGA gene; Caliber; Cardiovascular system; Catecholamines; Cell membrane; Cells; Characteristics; Chemistry; Chest; Chromaffin granule; Chromogranin A; Clinical; Collaborations; Deoxyglucose; Detection; Development; Diagnosis; Disease; Etiology; Family history of; Frequencies; Functional Imaging; Functional disorder; Gene Expression Profile; Gene Mutation; Genes; Genetic; Genetic Markers; Genomics; Germ-Line Mutation; Goals; Health Professional; Hepatic; High Prevalence; Hour; Hypertension; Image; In 111 Pentetreotide; Incidence; Injection of therapeutic agent; Institutes; Interdisciplinary Study; Knowledge; Laboratories; Lesion; Link; Liver; Liver neoplasms; Localized; Location; Low Prevalence; Magnetic Resonance Imaging; Malignant - descriptor; Malignant Neoplasms; Malignant Pheochromocytoma; Medical; Medical center; Messenger RNA; Metastatic Lesion; Metastatic Pheochromocytoma; Methods; Modality; Molecular; Molecular Biology; Molecular Genetics; Monitor; Morphologic artifacts; Motion; Mus; Mutation; Neoplasm Metastasis; Neuroendocrinology; Noise; Numbers; Organ; Paraganglioma; Pathogenesis; Pathway interactions; Patients; Peptides; Phenotype; Pheochromocytoma; Play; Positron-Emission Tomography; Primary Neoplasm; Proteins; Radionuclide Imaging; Rate; Reference Standards; Research; Research Personnel; Role; Scanning; Secretory Vesicles; Series; Signal Transduction; Staging; Symptoms; Syndrome; Technetium Tc 99m Medronate; Technology; Testing; Time; Tissues; Translational Research; Translations; United States National Institutes of Health; Vesicle; base; bone; clinical Diagnosis; clinical application; concept; driving force; improved; insight; interest; mouse model; neuropeptide Y; noradrenergic; patient oriented; patient oriented research; pentetreotide; radiofrequency; respiratory; secretory protein; size; subcutaneous; symposium; tumor; tumorigenesis; uptake

The Section is conducting patient-oriented research about the etiology, pathophysiology, genetics, diagnosis, and treatment of pheochromocytoma. 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. SDHB-related pheochromocytoma and paraganglioma SDHB mutations appear to be associated with more aggressive tumor behavior and a higher rate of malignancy. In our initial study of this area, we examined the frequency of SDHB mutations in patients with malignant pheochromocytomas/paragangliomas. Pathogenic SDHB mutations were found in 30% of these patients. In those patients who presented initially with primary abdominal paragangliomas, mutations of the SDHB gene were associated with about one half of all malignancies justifying a high priority for SDHB germline mutation testing in these patients. In a further study, we aimed to gain more detailed insight into the clinical and biochemical characteristics of SDHB-associated paragangliomas. Thirty patients with abdominal or thoracic paragangliomas and SDHB mutations were studied. At presentation, 21% of patients lacked any symptoms of catecholamine excess. Family history was positive for paraganglioma in only 10% of patients. Primary tumors were found in extra-adrenal locations in 97% of patients and had a mean diameter of about 8 cm. In 30% of patients, metastatic disease was already apparent at the initial diagnosis and 97% of patients eventually developed metastases after 2.64.1 years. The biochemical phenotype indicated hypersecretion of both NE and DA in about half of the patients. There was no correlation between the particular SDHB gene mutation type and clinical presentations, including malignant potential. The importance of functional imaging was demonstrated by an additional study. SDHB gene mutations carry a high malignant potential; thus, timely and accurate localization of SDHB related pheochromocytomas and paragangliomas is critical for implementing optimal treatment. Sensitivities for detection of metastases were compared between 18F-fluorodopamine (18F-FDA) and 18F-fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET), 123I- and 131I-metaiodobenzylguanidine (MIBG), 111In-Pentetreotide (Octreoscan) and Tc-99m-methylene diphosphonate (MDP) bone scintigraphy in 30 patients with SDHB-associated paragangliomas. Lesions detected by CT and MRI were used as a standard of reference. Sensitivity according to patient/body region were as follows: 123/131I-MIBG 57/68; Octreoscan: 59/81; 18F-FDA: 70/99; FDG: 97/100. At least 90% of regions that were false negative on 123I-MIBG scintigraphy or 18F-FDA PET were detected by FDG PET. We concluded that FDG PET is the preferred functional imaging modality for staging and treatment monitoring of SDHB-related metastatic paraganglioma. VHL- and MEN-related pheochromocytoma Pheochromocytomas in MEN 2 produce EPI; those in VHL syndrome produce NE. Patients with MEN 2 pheochromocytomas have a high incidence of paroxysmal attacks and have a higher prevalence of hypertension and other cardiovascular problems than do patients with VHL pheochromocytomas. Therefore, we hypothesized that these distinctions relate to differences in expression of the transporters responsible for uptake and storage of catecholamines, namely neuropeptide Y (NPY), a vasoactive peptide with influences on blood pressure, and of chromogranin A and B (CGA, CGB), major secretory proteins of chromaffin granules. We demonstrated that MEN 2-related pheochromocytomas expressed more cell membrane noradrenergic transporter mRNA and protein than tumors from VHL patients. This difference was associated with larger numbers of storage vesicles and higher tissue content of catecholamines in MEN 2 than in VHL tumors. In another study we found that tumor NPY levels in VHL patients were significantly lower than in those from MEN 2 patients for both mRNA and the peptide. We also showed that pheochromocytomas from MEN 2 patients expressed substantially more CGA and CGB than tumors from VHL patients at both the mRNA and protein level. We concluded that the differences in tumor CGA expression could contribute to differences in secretory vesicle formation and secretion in the two types of tumors. This may contribute, for example, to the lower prevalence of hypertension in VHL patients than in patients with MEN 2-related pheochromocytoma. We also evaluated seven VHL patients with adrenal pheochromocytomas using CT, MRI, 123/131I-MIBG scintigraphy, and 18F-FDA PET. We concluded that 18F-FDA PET in conjunction with CT/MRI should be considered an effective method for the proper localization of VHL-related adrenal pheochromocytoma. An animal model of pheochromocytoma As a continuation of our first anatomical studies, we have further optimized the use of microCT and MRI to localize organ and bone metastatic lesions in the mouse model of pheochromocytoma. To improve localization of liver lesions using microCT, we combined two time points of scanning. The first microCT scan was performed immediately after FenestraTMLC injection where both liver lesions and vessels were detected. The second scan was performed 3 hours after FenestraTMLC injection when the contrast disappeared from the vessels and allowed detection of the liver metastatic lesions. By combining these two scans we were able to precisely localize liver lesions at sizes of 300-500 ?m in diameter. To detect other organ lesions, we used MRI. To optimize the signal-to-noise ratio, we used a small animal dedicated radiofrequency coil on 3T MRI; and to reduce the motion artifacts, we applied respiratory triggering on anesthetized mice while scanning. In addition to developing and optimizing methods for tumor imaging in mice, we have also used the mouse model of metastatic pheochromocytoma to study the gene expression profile of hepatic and subcutaneous lesions derived from mouse pheochromocytoma cells. It is of great interest to develop biomarkers or genomic screens that may predict the aggressiveness of pheochromocytoma. We therefore used the mouse model of metastatic pheochromocytoma to study the gene expression profile of hepatic and subcutaneous lesions. Comparison of subcutaneous and liver tumors revealed 8 genes (Pten, Mdm2, Metap2, Rb1, Reck, S100a4, Timp2 and Timp3) with 2-fold increases in expression in the liver compared to subcutaneous tumors. QT-PCR analysis confirmed 5 of these genes. This study provides initial information about which genes could play an important role in the aggressive behavior of metastatic pheochromocytoma cells. This study provides initial information about which genes could play an important role in the aggressive behavior of metastatic pheochromocytoma cells. In 2006, we organized the 1st SDHB Related Pheochromocytoma conference as a series of interactive sessions for health professionals and patients. This year we are organizing the Pheochromocytoma 2007 conference.

该科正在以患者为中心开展有关嗜铬细胞瘤的病因、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。 SDHB相关的嗜铬细胞瘤和副神经节瘤 SDHB 突变似乎与更具侵袭性的肿瘤行为和更高的恶性肿瘤发生率相关。在我们对该领域的初步研究中，我们检查了恶性嗜铬细胞瘤/副神经节瘤患者的 SDHB 突变频率。其中 30% 的患者发现了致病性 SDHB 突变。在那些最初患有原发性腹部副神经节瘤的患者中，SDHB 基因突变与大约一半的恶性肿瘤相关，这证明对这些患者进行 SDHB 种系突变检测具有高度优先性。 在进一步的研究中，我们旨在更详细地了解 SDHB 相关副神经节瘤的临床和生化特征。对 30 名患有腹部或胸部副神经节瘤和 SDHB 突变的患者进行了研究。就诊时，21% 的患者没有任何儿茶酚胺过量的症状。只有 10% 的患者有副神经节瘤家族史。 97%的患者原发性肿瘤发现于肾上腺外位置，平均直径约为8厘米。 30% 的患者在初次诊断时已出现明显的转移性疾病，97% 的患者最终在 2.64.1 年后出现转移。生化表型表明约一半患者的 NE 和 DA 分泌过多。特定的 SDHB 基因突变类型与临床表现（包括恶性潜能）之间没有相关性。 另一项研究证明了功能成像的重要性。 SDHB基因突变具有高度恶性潜力；因此，及时准确地定位SDHB相关的嗜铬细胞瘤和副神经节瘤对于实施最佳治疗至关重要。比较了 18F-氟多巴胺 (18F-FDA) 和 18F-氟-2-脱氧-D-葡萄糖 (FDG) 正电子发射断层扫描 (PET)、123I- 和 131I-间碘苄基胍 (MIBG)、111In-戊曲肽 (Octreoscan) 和 Tc-99m-亚甲基之间的转移检测灵敏度 二磷酸盐 (MDP) 骨闪烁扫描对 30 名 SDHB 相关副神经节瘤患者进行了研究。 CT和MRI检测到的病变作为参考标准。根据患者/身体区域的敏感性如下：123/131I-MIBG 57/68；奥克斯特扫描：59/81； 18F-FDA：70/99； FDG：97/100。 FDG PET 检测到至少 90% 的 123I-MIBG 闪烁扫描或 18F-FDA PET 假阴性区域。我们的结论是，FDG PET 是 SDHB 相关转移性副神经节瘤分期和治疗监测的首选功能成像方式。 VHL 和 MEN 相关嗜铬细胞瘤 MEN 2 中的嗜铬细胞瘤产生 EPI；那些患有VHL综合征的人会产生NE。与 VHL 嗜铬细胞瘤患者相比，MEN 2 嗜铬细胞瘤患者阵发性发作的发生率较高，高血压和其他心血管问题的患病率也较高。因此，我们假设这些差异与负责儿茶酚胺摄取和储存的转运蛋白的表达差异有关，即神经肽 Y (NPY)（一种影响血压的血管活性肽）以及嗜铬粒蛋白 A 和 B (CGA、CGB)（嗜铬颗粒的主要分泌蛋白）的表达差异。我们证明，MEN 2 相关的嗜铬细胞瘤比 VHL 患者的肿瘤表达更多的细胞膜去甲肾上腺素转运蛋白 mRNA 和蛋白质。这种差异与 MEN 2 中比 VHL 肿瘤中更多的储存囊泡和更高的儿茶酚胺组织含量有关。在另一项研究中，我们发现 VHL 患者的肿瘤 NPY mRNA 和肽水平均显着低于 MEN 2 患者。我们还表明，MEN 2 患者的嗜铬细胞瘤在 mRNA 和蛋白质水平上比 VHL 患者的肿瘤表达更多的 CGA 和 CGB。我们得出的结论是，肿瘤 CGA 表达的差异可能导致两类肿瘤中分泌囊泡形成和分泌的差异。 例如，这可能导致 VHL 患者的高血压患病率低于 MEN 2 相关嗜铬细胞瘤患者。 我们还使用 CT、MRI、123/131I-MIBG 闪烁扫描和 18F-FDA PET 对 7 名患有肾上腺嗜铬细胞瘤的 VHL 患者进行了评估。 我们的结论是，18F-FDA PET 结合 CT/MRI 应被视为正确定位 VHL 相关肾上腺嗜铬细胞瘤的有效方法。 嗜铬细胞瘤动物模型 作为我们首次解剖学研究的延续，我们进一步优化了 microCT 和 MRI 的使用，以定位嗜铬细胞瘤小鼠模型中的器官和骨转移灶。为了利用 microCT 提高肝脏病变的定位，我们结合了两个时间点的扫描。注射 FenestraTMLC 后立即进行第一次 microCT 扫描，检测到肝脏病变和血管。第二次扫描在 FenestraTMLC 注射后 3 小时进行，此时造影剂从血管中消失，从而可以检测到肝脏转移病灶。通过结合这两次扫描，我们能够精确定位直径为 300-500 微米的肝脏病变。为了检测其他器官病变，我们使用了 MRI。为了优化信噪比，我们在3T MRI上使用了小动物专用射频线圈；为了减少运动伪影，我们在扫描时对麻醉小鼠应用呼吸触发。 除了开发和优化小鼠肿瘤成像方法外，我们还使用转移性嗜铬细胞瘤小鼠模型来研究小鼠嗜铬细胞瘤细胞来源的肝脏和皮下病变的基因表达谱。开发可以预测嗜铬细胞瘤侵袭性的生物标志物或基因组筛选非常有意义。因此，我们使用转移性嗜铬细胞瘤小鼠模型来研究肝脏和皮下病变的基因表达谱。皮下肿瘤和肝脏肿瘤的比较显示，与皮下肿瘤相比，8 个基因（Pten、Mdm2、Metap2、Rb1、Reck、S100a4、Timp2 和 Timp3）在肝脏中的表达量增加了 2 倍。 QT-PCR 分析证实了其中 5 个基因。这项研究提供了有关哪些基因在转移性嗜铬细胞瘤细胞的侵袭行为中发挥重要作用的初步信息。这项研究提供了有关哪些基因在转移性嗜铬细胞瘤细胞的侵袭行为中发挥重要作用的初步信息。 2006 年，我们组织了第一届 SDHB 相关嗜铬细胞瘤会议，为卫生专业人员和患者举办了一系列互动会议。今年我们将组织 2007 年嗜铬细胞瘤会议。