Thrombotic Disorder Banking Core

虽然在血栓性疾病和血栓性微血管病的病因学的理解方面已经取得了进展，但大多数患病患者没有确定的突变或诱发风险因素。为了更好地了解血栓性疾病和血栓性微血管病的分子缺陷，未来的工作将必须确定存在于DNA中的突变和血浆蛋白的特异性活性。因此，将建立一个核心，以便于从合格患者中采集血液样本。该核心将用于存档血栓性微血管病和其他血栓性疾病患者的血浆和DNA样本，并将在华盛顿大学现有和成功的转化病理学和分子表型核心（TPMPC）的参数范围内运行。 核心还将建立和维护一个数据库，该数据库将采集相关的流行病学、疾病相关、预后、治疗和结局数据，并与相应的库存血浆和DNA样本进行去识别化关联。 血栓性疾病库核心将负责确定符合样本采集条件的患者，获得合格受试者的知情同意书，采集血液并使用血液样本获得血浆和从血液样本中的白色血细胞分离的DNA。样本采集的入选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、非典型溶血性尿毒综合征和与血栓性血小板减少性紫癜相关的HELLP综合征。 项目1和2研究的妊娠。将在诊断时采集血样，并在门诊随访期间再次采集血样。 核心是必不可少的个人研究项目，因为它将提供DNA的补体途径蛋白，凝血途径蛋白，和其他候选基因可能参与血栓性微血管病的发病机制的整个外显子组测序。具体而言，这一核心将是工具性的， 本申请中项目1和项目2的目的。 存档的血浆和血清样本对于项目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。来自对照患者和患有血栓性血小板减少性紫癜（TTP）的患者的血浆将用于开发ADAMTS13活性的快速和灵敏的测定， ADAMTS13的抑制剂。此外，血浆和DNA样本将用于临床表型分析和特定治疗的评价。样本还将用于表征与先前未识别的血栓性微血管病机制相关的缺陷。 项目2（血栓性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（阿胡斯）（一种血栓性微血管病）中其他补体和凝血蛋白的突变。迄今为止，在阿胡斯患者中已经鉴定了五种补体蛋白和止血调节剂的突变。因为只有70%的患者发现了突变， 需要对另外30%的阿胡斯患者的缺陷进行表征。也有数据表明，这些蛋白质的突变也可能存在于抗磷脂综合征和妊娠HELLP综合征患者中。 我们将利用华盛顿大学Mark沃森博士领导的临床和转化科学研究所现有转化病理学和分子表型核心（TPMPC）的服务。核心将负责冷冻保存血样中的血浆， 白色血细胞，用于从白色血细胞中纯化DNA和用于数据管理。在签署知情同意书时，将为血样分配一个唯一的、随机生成的6位数患者识别号。带有六位数字的标签将用于血液样本和纸质主列表，其中包含患者姓名、病历编号、出生日期和知情同意日期。总名单将保存在一个上锁的保险箱里。这个保险箱的钥匙将由研究协调员、核心主任和主要研究者持有。血液样本将在2小时内手动运送至组织采集实验室。此外，将在血液学部门处理并储存一式两份样本，以确保样本完整性。 转化病理学和分子表型核心已被许多小组成功使用，包括华盛顿大学的急性白血病小组和乳腺癌小组，用于前瞻性收集和处理与临床试验相关的人类血液和组织标本。

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.