Studies in the Pathogenesis of Systemic Capillary Leak Syndrome

全身毛细血管渗漏综合征发病机制的研究

基本信息

批准号：
8946466
负责人：
Kirk m Druey
金额：
$ 61.43万
依托单位：
NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
依托单位国家：
美国
项目类别：
财政年份：
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/8946466
关键词：
2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole 3p25.3 Actins Acute Address Anaphylaxis Angioneurotic Edema Angiopoietin-2 Antigen Targeting Antihypertensive Agents B-Cell Neoplasm Binding Biological Assay Biological Markers Blood Blood Cells Blood Vessels Blood capillaries Bradykinin Candidate Disease Gene Capillary Leak Syndrome Chromosomes, Human, Pair 6 Clinical Clinical Protocols Data Development Disease Disease remission Edema Endothelial Cells Etiology Event Extravasation Functional disorder Gene Expression Profile Generations Genetic Genomic DNA Goals Growth and Development function Hematopoietic Histamine Human Hypersensitivity Hypotension Hypovolemia IL8 gene Immune Immunoglobulin G Immunoglobulins In Vitro Inbred Strains Mice Intravenous Immunoglobulins Link Liquid substance Maps Mass Spectrum Analysis Mediating Mediator of activation protein Molecular Monoclonal gammopathy of uncertain significance Morbidity - disease rate Multiple Myeloma Mus Nature Oliguria Paraproteins Pathogenesis Pathology Patients Permeability Phase Phenotype Plasma Plasma Cells Plasmacytic Leukemia Polymorphism Analysis Population Predisposition Premalignant Process Property Proteins Proteome Protocols documentation RNA Reporting Research Resistance SJL Mouse Sepsis Serotonin Serum Shock Signs and Symptoms Single Nucleotide Polymorphism Source Stress Fibers Symptoms Syndrome Tissues Tumor Necrosis Factor-alpha Two-Dimensional Gel Electrophoresis United States National Institutes of Health Ursidae Family Validation Vascular Endothelial Growth Factors Xenograft Model anasarca angiogenesis base cadherin 5 capillary cell type chemotherapy cohort cytokine exome experience genetic association genome sequencing genome-wide human TNF protein macromolecule mortality novel therapeutic intervention response

项目摘要

Immune dysregulation may contribute to the pathophysiologic findings seen in SCLS. A monoclonal gammopathy of unknown significance (MGUS, which is a premalignant precursor to multiple myeloma (MM), in which a clonal plasma cell population secretes monoclonal immunoglobulin (Ig, also referred to as a paraprotein) detectable in patient sera, is present in a majority of SCLS cases. Several patients with SCLS in whom MGUS evolved into frank myeloma or plasma cell leukemia experienced fewer capillary leak episodes after chemotherapy for their hematopoietic disorder. These findings suggest that the monoclonal paraprotein from the dysregulated plasma cell population may be the direct or indirect source of the pathophysiologic findings observed. We are exploring the molecular mechanisms of SCLS by examining the function of the monoclonal Ig in the development of vascular pathology in vitro. Using monoclonal Ig from SCLS patients, we will determine whether it binds to a specific cell type, target antigen (if any), and resulting effect (direct or indirect) on endothelial cells. Using 2D gel electrophoresis and mass spectrometry, we have identified proteins that appear to bind to IgG from subjects with SCLS, but not healthy controls. Confirmatory studies of potential paraprotein targets are underway. We are also characterizing the transcriptome of blood cell RNA and the proteome of SCLS serum/plasma, both pre- and post-attack, to determine whether specific biomarkers of acute symptoms can be identified. We have now evaluated nearly 40 patients at the NIH clinical center under this protocol in the last 5 years. We are the primary worldwide referral center for research on SCLS. The transient episodes of hypotensive shock and anasarca in SCLS are thought to arise from reversible microvascular barrier dysfunction. Application of episodic SCLS sera, but neither the purified immunoglobulin fraction nor sera obtained from patients during remission, to normal human microvascular endothelial cells caused vascular endothelial cadherin internalization, disruption of interendothelial junctions, actin stress fiber formation, and increased permeability in complementary functional assays. Circulating permeability factors, vascular endothelial growth factor (VEGF) and angiopoietin 2 (Ang2), were elevated in episodic SCLS sera but not in remission sera. These results suggest that humoral factors such as VEGF and Ang2 contribute to transient endothelial contraction in SCLS, suggesting a molecular mechanism for this highly lethal disorder. Current studies are aimed at examined responses of endothelial cells derived from subjects with SCLS and controls to permeability mediators. In FY 2014, we characterized a subject with atypical disease features using detailed mechanistic studies. Microvascular barrier function assays corroborated our atypical patient's clinical resistance to IVIG and suggested that his episodes were driven by acute phase cytokines IL-8 and TNF-alpha. Thus, our data suggest that SCLS may have clinically varying forms of presentation and that within the group of patients with SCLS, different cytokines may mediate capillary leak. In FY 2014, using Affymetrix Single Nucleotide Polymorphism (SNP) microarrays, we performed the first genome-wide SNP analysis of SCLS in a cohort of 12 disease subjects and 18 controls. Exome capture sequencing was performed on genomic DNA from nine of these patients as validation for the SNP-chip discoveries and de novo data generation. We identified candidate susceptibility loci for SCLS. An inbred mouse strain was identified (SJL/J), whose phenotype bears similarity to SCLS, including spontaneous vascular hypersensitivity to a variety of permeability factors including histamine (HA), serotonin, and bradykinin, important mediators of endothelial hyperpermeability in humans, and B cell neoplasms characterized by the presence of serum paraproteins. A recessive locus in SJL mice controlling systemic vascular hypersensitivity to HA (Shs) was mapped to an interval on mouse chromosome 6 syntenic with human 3p25.3, which harbors the primary genetic association with SCLS in humans. This interval contains several gene candidates, which will be analyzed in the SCLS cohort.

免疫失调可能有助于SCL中看到的病理生理发现。具有未知意义的单克隆性γ（MGU，是多发性骨髓瘤（MM）的前体前体（MG），其中克隆浆细胞群体分泌单克隆免疫球蛋白（Ig）（Ig，Ig，也称为paraprotein）在SCL中均可在SCL中检测到多个患者，以此为大多数患者。骨髓瘤或血浆细胞白血病的造血疾病后，毛细血管泄漏发作较少。我们正在通过检查单克隆Ig在体外血管病理发展中的功能来探索SCL的分子机制。使用SCLS患者的单克隆IG，我们将确定它是否与特定细胞类型结合，靶抗原（如果有）以及对内皮细胞的产生影响（直接或间接）。使用2D凝胶电泳和质谱法，我们发现了似乎与具有SCLS受试者但没有健康对照受试者的IgG结合的蛋白质。潜在的副蛋白靶标的验证性研究正在进行中。我们还表征了血细胞RNA的转录组和攻击前和攻击后SCLS血清/血浆的蛋白质组，以确定是否可以鉴定出急性症状的特定生物标志物。在过去的5年中，根据该方案，我们在NIH临床中心评估了近40名患者。我们是SCLS研究的主要全球转诊中心。 SCL中降压冲击和Anasarca的短暂发作被认为是由可逆的微血管屏障功能障碍引起的。发作性SCLS血清的应用，但在缓解过程中纯化的免疫球蛋白分数和从患者中获得的血清均未导致人类微血管内皮细胞引起血管内皮内皮核蛋白内在化，破坏内皮间固体结合的破坏，肌动蛋白压力纤维形成，肌动蛋白的形成，辅助功能性疾病的持久性增加。循环渗透性因子，血管内皮生长因子（VEGF）和血管生成素2（ANG2）在情节性SCLS血清中升高，但在缓解血清中却没有升高。这些结果表明，诸如VEGF和ANG2之类的体液因素有助于SCL中的短暂内皮收缩，这表明这种高度致命的疾病的分子机制。当前的研究旨在检查来自具有SCL和对照对渗透性介质的受试者的内皮细胞的反应。在2014财年，我们使用详细的机械研究表征了具有非典型疾病特征的受试者。微血管屏障功能分析证实了我们非典型患者对IVIG的临床耐药性，并表明他的发作是由急性相细胞因子IL-8和TNF-Alpha驱动的。因此，我们的数据表明，SCL可能具有临床上不同的表现形式，并且在SCLS患者组中，不同的细胞因子可能介导毛细血管泄漏。在2014财年，使用Affymetrix单核苷酸多态性（SNP）微阵列，我们在12个疾病受试者和18个对照组中对SCLS进行了首次全基因组SNP分析。外外捕获测序是对来自这些患者的9名患者的基因组DNA进行的，作为SNP-CHIP发现和从头数据生成的验证。我们确定了SCLS的候选敏感性基因座。 An inbred mouse strain was identified (SJL/J), whose phenotype bears similarity to SCLS, including spontaneous vascular hypersensitivity to a variety of permeability factors including histamine (HA), serotonin, and bradykinin, important mediators of endothelial hyperpermeability in humans, and B cell neoplasms characterized by the presence of serum paraproteins.控制系统性血管超敏性HA（SHS）的SJL小鼠中的隐性基因座，将其与人类3P25.3同步的小鼠染色体6偶然地映射到一个间隔，后者将其与人类中的SCL相关。此间隔包含几个基因候选物，将在SCLS队列中进行分析。