Expression Profiling In Acute and Chronic Cardiac Allograft Rejection

急性和慢性心脏同种异体移植排斥反应中的表达谱

• 批准号: 8565288
• 负责人: Michael Solomon
• 金额: --
• 依托单位: CLINICAL CENTER
• 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/8565288
• 关键词: Acute; Allograft Tolerance; Allografting; Bioinformatics; Biological Markers; Biopsy; Blood Vessels; Cardiac; Cardiac Catheterization Procedures; Chronic; Clinical; Diagnosis; Diagnostic tests; Dyspnea; Echocardiography; Electrocardiogram; Enrollment; Fatigue; Future; Gene Expression Profile; Genes; Gold; Heart Transplantation; Heart failure; Histopathology; Image; Immunohistochemistry; Immunologics; Individual; Institutional Review Boards; Investigation; Laboratories; Laboratory Procedures; Low Grade Fever; Methodology; Methods; Microarray Analysis; Molecular Profiling; Morbidity - disease rate; Myocardial Infarction; Nuclear; Oligonucleotide Microarrays; Pathogenesis; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phase; Phosphorus; Procedures; Process; Proteomics; Protocols documentation; RNA; Regulatory Pathway; Research Infrastructure; Reverse Transcriptase Polymerase Chain Reaction; Risk; Sampling; Source; Spectrum Analysis; Survivors; Symptoms; T-Lymphocyte; Techniques; Testing; Therapeutic immunosuppression; Translating; Transplant Recipients; Transplantation; Ultrasonography; Universities; Vascular Diseases; Ventricular; Western Blotting; allograft rejection; base; density; diagnosis standard; drug development; experience; functional genomics; heart allograft; mortality; programs; success; sudden cardiac death; tool

Acute cardiac allograft cellular rejection remains a significant source of morbidity and mortality within the first year after heart transplantation. Afterwards, cardiac allograft vasculopathy (CAV), as a result of chronic vascular rejection, is the major cause of morbidity and mortality. Within the first year post-transplantation, almost two-thirds of recipients will experience at least one rejection episode. At five years post-transplantation, nearly 50% of survivors will have CAV. In the first year after transplantation, nearly 63% of patients experience at least one episode of cardiac rejection and approximately one third of these patients will have multiple episodes. The clinical symptoms of acute cardiac rejection are relatively nonspecific (fatigue, dyspnea, low grade fever). Most CAV patients remain asymptomatic until they develop serious problems such as myocardial infarction, heart failure, ventricular dysrhythmias or sudden cardiac death. No widely accepted noninvasive method exists for the accurate diagnosis of acute or chronic cardiac rejection. Noninvasive methods such as electrocardiography, echocardiography, and nuclear studies all have been studied, but have been unsuccessful, thus far, for either condition. Several methodologies have been studied including electrocardiography, echocardiography, nuclear imaging, and phosphorus spectroscopy without success. The current gold standard for the diagnosis of acute cellular rejection remains right ventricular endomyocardial biopsy. This is an invasive method of diagnosis subject to morbidity and random sampling and interpretation error. Likewise, the gold standard for diagnosing CAV is cardiac catheterization with intravascular ultrasound, an invasive procedure also subject to morbidity. We are applying functional genomics to study acute cardiac allograft cellular rejection and CAV. Our group has developed and tested standard laboratory procedures for sample processing and if necessary amplification. We have established laboratory and bioinformatics infrastructure to support oligonucleotide microarray investigations. Two major local transplant programs (Johns Hopkins University and INOVA-Fairfax) have been or are currently collaborating. We have an IRB approved protocol and are currently actively enrolling patients (184 individuals enrolled to date). We hypothesize that large scale expression profiling of circulating peripheral blood mononuclear cells (PBMC, predominantly T lymphocytes) will identify genes that can serve as reliable biomarkers of acute and chronic cardiac cellular rejection. In the initial bench phase of the project, PBMCs are obtained from heart transplant recipients during periods of immunological tolerance of the allograft (no acute rejection) and immunologic intolerance of the allograft (acute rejection) and from heart transplant recipients with and without CAV to determine whether unique gene expression patterns are associated with each state. Other analytic tools that may be employed include proteomics, RT-PCR, Western blot, insitu-hybridization, immunohistochemistry, and histopathology. In the latter phase of the project we hope to translate these profiles into an acceptable bedside test for acute and chronic cardiac allograft cellular rejection. In addition to developing a biomarker approach to the diagnosis of rejection in cardiac transplant patients, expression profiling has the potential to identify immunoregulatory pathways that can serve as new targets for immunosuppressive therapy (rational drug development). In 2010, we processed 64 samples for high density oligonucleotide microarray analysis. In the latter part of 2010, a second batch containing 168 samples was processed to TRNA, the required step before going to microarrays. In 2011, an additional 139 samples were processed to TRNA and 89 samples were further processed to high density oligonucleotide microarray analysis. Since October 2011 through August 2012 we have processed an additional 44 samples to TRNA and processed 397 TRNA samples to an RNA stable state. The protocol continues to actively enroll subjects.

心脏移植后第一年，急性心脏同种异体移植细胞排斥仍然是发病率和死亡率的重要来源。之后，由于慢性血管排斥反应而导致心脏同种异体血管病（CAV）是发病率和死亡率的主要原因。 转移后的第一年，几乎三分之二的接收者将至少经历一个拒绝情节。 移植后五年，将近50％的幸存者将拥有CAV。在移植后的第一年，将近63％的患者至少经历了心脏排斥的至少一集，其中大约三分之一的患者会发生多个发作。急性心脏排斥的临床症状是相对非特异性的（疲劳，呼吸困难，低级发烧）。大多数CAV患者一直无症状，直到他们出现严重的问题，例如心肌梗死，心力衰竭，心室心律失常或心脏猝死。 对于准确诊断急性或慢性心脏排斥，没有广泛接受的非侵入性方法。诸如心电图，超声心动图和核研究之类的无创方法都已进行了研究，但到目前为止，对于这两种情况都没有成功。 已经研究了几种方法，包括心电图，超声心动图，核成像和磷光谱法而没有成功。当前诊断急性细胞排斥反应的金标准仍然是右心室内膜活检。 这是一种侵入性的诊断方法，受发病率和随机抽样和解释误差的影响。 同样，用于诊断CAV的金标准是心脏导管插入术，具有血管内超声，这是一种侵入性手术，也需要发病。 我们正在应用功能基因组学来研究急性心脏同种异体移植细胞排斥和CAV。我们的小组已经制定并测试了标准实验室程序，以进行样品处理和必要时的放大。 我们已经建立了实验室和生物信息学基础设施，以支持寡核苷酸微阵列调查。 目前或目前正在合作的两个主要本地移植计划（约翰·霍普金斯大学和Inova-Fairfax）。 我们有一个IRB批准的方案，目前正在积极招募患者（迄今为止有184名个人入学）。 我们假设循环外周血单核细胞（PBMC，主要是T淋巴细胞）的大规模表达分析将鉴定可以用作急性和慢性心脏细胞排斥的可靠生物标志物的基因。在项目的初始基准阶段中，PBMC是在同种异体移植物免疫耐受性（无急性排斥）和同种异体移植物（急性抑制）的免疫学不耐受性的情况下从心脏移植受者获得的，并且是从心脏移植受者和无CAV的情况下确定独特的基因表达模式是否与每个状态相关的。 可以使用的其他分析工具包括蛋白质组学，RT-PCR，Western印迹，腹膜杂交，免疫组织化学和组织病理学。 在该项目的后期，我们希望将这些曲线转化为可接受的床边测试，以进行急性和慢性心脏同种异体移植细胞排斥反应。除了开发一种生物标志物方法来诊断心脏移植患者的排斥反应外，表达分析还具有鉴定免疫调节途径的潜力，可以作为免疫抑制治疗（理性药物开发）的新靶标。 2010年，我们处理了64个样品，用于高密度寡核苷酸微阵列分析。 在2010年的下半年，将第二批包含168个样品处理到tRNA，这是前往微阵列之前所需的步骤。 2011年，将另外139个样品处理到tRNA中，并将89个样品进一步处理至高密度寡核苷酸微阵列分析。 自2011年10月至2012年8月以来，我们已经处理了44个样品，并将397个tRNA样品处理到RNA稳定状态。 该协议继续积极注册受试者。