Thrombotic Disorder Banking Core

虽然对血栓性疾病和血栓性微血管病变的病因的了解已经取得了进展，但大多数患者没有已确定的突变或易感危险因素。为了更好地了解血栓性疾病和血栓性微血管病变中的分子缺陷，未来的工作将不得不确定DNA中存在的突变和血浆蛋白的特定活性。因此，我们会设立一个中心，以便从合资格的病人身上收集血液样本。该核心将对血栓性微血管病和其他血栓性疾病患者的血浆和DNA样本进行存档，并将在华盛顿大学现有的、成功的翻译病理学和分子表型核心(TPMPC)的参数范围内运行。 该核心还将建立和维护一个数据库，该数据库将收集相关的流行病学、疾病相关、预后、治疗和结果数据，并与相应的银行血浆和DNA样本建立未查明的联系。 血栓性疾病银行核心将负责确定有资格采集样本的患者、获得符合条件的参与者的知情同意、采集血液并使用血液样本获取血浆和从血液样本中的白细胞中分离出的DNA。样本收集的纳入标准将包括血栓性微血管病的诊断，如血栓性血小板减少性紫癜、非典型溶血性尿毒症综合征和HELLP综合征与 项目1和2中的研究中的怀孕。将在诊断时和门诊随访期间再次采集血样。 核心对单个研究项目至关重要，因为它将为补体途径蛋白、凝血途径蛋白和其他可能参与血栓性微血管病变发病机制的候选基因的整个外显子组测序提供DNA。具体地说，这个核心将是有益的 符合本申请中项目1和2的目标。 存档的血浆和血清样本将对项目1(血栓性微血管病的病理生理学和治疗(SADLER))的成功至关重要，该项目旨在了解血栓性微血管病的病理生理学和治疗。来自对照患者和血栓性血小板减少性紫癜(TTP)患者的血浆将被用来建立快速和敏感的ADAMTS13活性和 ADAMTS13的抑制剂。此外，血浆和DNA样本将用于临床表型和特定治疗的评估。样本也将被用来表征与以前未知的血栓性微血管病变机制相关的缺陷。 项目2(血栓微血管病的遗传易感性(Atkinson))将确定非典型溶血性尿毒症综合征(AHUS)中其他补体和凝血蛋白的突变，这是一种血栓性微血管病。到目前为止，已经在aHUS患者中发现了五种补体蛋白和一种止血调节因子的突变。因为只有70%的患者发现了突变，而且 需要开展工作来确定其他30%的aHUS患者的缺陷。也有数据表明，这些蛋白的突变也可能存在于那些患有抗磷脂综合征和妊娠HELLP综合征的人中。 我们将利用华盛顿大学马克·沃森博士领导的临床和翻译科学研究所现有的翻译病理学和分子表型核心(TPMPC)的服务。该核心将负责从血液样本中冷冻保存血浆，以分离 白血球，用于从白血球中提纯DNA和数据管理。血样将在同意时被分配一个唯一的、随机生成的六位数的患者识别码。血液样本和包含患者S姓名、病历编号、出生日期和同意日期的纸质主名单上将使用带有六位数字的标签。总名单将保存在一个上了锁的保险箱里。这个保险箱的钥匙将由研究协调员、核心主任和主要调查人员掌握。血样将在2小时内被手送到组织采购实验室。此外，复制的样本将被处理并存储在血液科，以确保样本的完整性。 包括华盛顿大学的急性白血病组和乳腺癌组在内的许多小组已经成功地使用了翻译病理学和分子表型分析核心，用于与临床试验相关的人类血液和组织样本的预期收集和处理。

Although advances have been made in the understanding of etiologies of thrombotic disorders and thrombotic microangiopathies, the majority of afflicted patients do not have an identified mutation or predisposing risk factor. To better understand the molecular defects in thrombotic disorders and thrombotic microangiopathies, future work will have to identify mutations that exist in DNA and the specific activity of plasma proteins. Therefore, a core will be established to facilitate the collection of blood samples from eligible patients. This core will function to archive plasma and DNA samples from patients with thrombotic microangiopathies and other thrombotic disorders and will operate within the parameters of the existing and successful Translational Pathology and Molecular Phenotyping Core (TPMPC) at Washington University. The core will also establish and maintain a database that will capture relevant epidemiological, disease-related, prognostic, therapy, and outcomes data with de-identified linkage to corresponding banked plasma and DNA samples. The Thrombotic Disorder Banking Core will be responsible for identifying patients eligible for sample collection, obtaining informed consent from eligible participants, collecting blood and using the blood samples to obtain plasma and DNA isolated from white blood cells within the blood sample. Inclusion criteria for the collection of samples will include the diagnosis of a thrombotic microangiopathy such as thrombotic thrombocytopenic purpura, atypical hemolytic uremic syndrome, and HELLP syndrome associated with pregnancy for the studies in Project 1 and 2. Blood samples will be taken at diagnosis and again during outpatient follow-up. The core is essential to individual research projects because it will provide DNA for whole exome sequencing of complement pathway proteins, coagulation pathway proteins, and other candidate genes likely to be involved in the pathogenesis of thrombotic microangiopathies. Specifically, this core will be instrumental to the aims of Projects 1 and 2 in this application. Archived plasma and serum samples will be critical to the success of Project 1 (Pathophysiology and Treatment of Thrombotic Microangiopathy (Sadler)) that seeks to understand the pathophysiology and treatment of thrombotic microangiopathies. Plasma from control patients and those with thrombotic thrombocytopenic purpura (TTP) will be used to develop rapid and sensitive assays of ADAMTS13 activity and inhibitors of ADAMTS13. Furthermore, plasma and DNA samples will be used for clinical phenotyping and for the evaluation of specific treatments. Samples will also be used to characterize defects associated with previously unrecognized mechanisms of thrombotic microangiopathy. Project 2 (Genetic Predisposition to the Thrombomicroangiopathies (Atkinson)) will identify mutations in other complement and coagulation proteins in atypical hemolytic uremic syndrome (aHUS), a type of thrombotic microangiopathy. To date, mutations in five complement proteins and in a regulator of hemostasis have been identified in patients with aHUS. Because only 70% of patients have identified mutations, further work is needed to characterize the defect in the other 30% of aHUS patients. There is also data to suggest that mutations in these proteins may also be present in those with antiphospholipid syndrome and the HELLP syndrome of pregnancy. We will utilize the services of the existing Translational Pathology and Molecular Phenotyping Core (TPMPC) within the Institute of Clinical and Translational Sciences headed by Dr. Mark Watson at Washington University. The core will be responsible for cryopreservation of plasma from the blood samples, for isolation of white blood cells, for purification of DNA from the white blood cells and for data management. Blood samples will be assigned a unique, randomly-generated six digit patient identification number at the time of consent. A label with the six digit number will be used on the blood samples and on a paper master list that contains the patient¿s name, medical record number, date of birth and date of consent. The master list will be kept in a locked safe. The key to this safe will be held by the research coordinator, the core director, and the principal investigators. The blood samples will be hand carried to the tissue procurement lab within 2 hours. In addition, duplicate samples will be processed and stored within the hematology division to insure sample integrity. The Translational Pathology and Molecular Phenotyping Core has been used successfully by many groups including an acute leukemia group and a breast cancer group at Washington University for the prospective collection and processing of human blood and tissue specimens associated with clinical trials.