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

• 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
• 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

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.

免疫功能失调可能导致SCLS的病理生理学改变。意义不明的单克隆丙种球蛋白病（MGUS）是多发性骨髓瘤（MM）的癌前病变，其中克隆浆细胞群分泌可在患者血清中检测到的单克隆免疫球蛋白（IG，也称为副蛋白），存在于大多数SCLS病例中。 几名MGUS演变为明显骨髓瘤或浆细胞白血病的SCLS患者在化疗后发生的毛细血管渗漏事件较少。 这些结果表明，单克隆副蛋白从失调的浆细胞群体可能是直接或间接的来源，观察到的病理生理学结果。 我们正在通过检测单克隆IG在体外血管病理学发展中的作用来探索SCLS的分子机制。使用SCLS患者的单克隆IG，我们将确定其是否与特定细胞类型、靶抗原（如果有）结合，以及对内皮细胞的影响（直接或间接）。使用2D凝胶电泳和质谱，我们已经确定了似乎与SCLS受试者的IgG结合的蛋白质，但不是健康对照。潜在副蛋白靶点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SCLS血清/血浆的蛋白质组，包括发作前和发作后，以确定是否可以识别急性症状的特定生物标志物。在过去的5年里，我们已经在NIH临床中心根据该方案评估了近40例患者。我们是SCLS研究的主要全球转诊中心。 SCLS中短暂发作的水肿性休克和全身水肿被认为是由可逆性微血管屏障功能障碍引起的。在补充功能测定中，将发作性SCLS血清应用于正常人微血管内皮细胞引起血管内皮钙粘蛋白内化、内皮间连接破坏、肌动蛋白应力纤维形成和渗透性增加，但既不应用纯化的免疫球蛋白组分也不应用从缓解期患者获得的血清。循环通透性因子血管内皮生长因子（VEGF）和血管生成素2（Ang 2）在SCLS发作期血清中升高，但在缓解期血清中未升高。这些结果表明，体液因子，如VEGF和Ang 2有助于短暂的内皮细胞收缩SCLS，提示这种高致死性疾病的分子机制。目前的研究旨在检查来自SCLS受试者和对照组的内皮细胞对渗透性介质的反应。 在2014财年，我们使用详细的机制研究描述了具有非典型疾病特征的受试者。微血管屏障功能测定证实了我们的非典型患者对IVIG的临床耐药性，并表明他的发作是由急性期细胞因子IL-8和TNF-α驱动的。因此，我们的数据表明，SCLS可能有临床上不同形式的介绍，并与SCLS患者组内，不同的细胞因子可能介导毛细血管渗漏。 在2014财年，我们使用Affyscore单核苷酸多态性（SNP）微阵列，在12名疾病受试者和18名对照者的队列中进行了SCLS的首次全基因组SNP分析。对这些患者中的9名的基因组DNA进行外显子组捕获测序，作为SNP芯片发现和从头数据生成的验证。我们确定了SCLS的候选易感基因位点。确定了一种近交系小鼠品系（SJL/J），其表型与SCLS相似，包括对多种渗透性因子（包括组胺（HA）、5-羟色胺和缓激肽（人体内皮细胞高渗透性的重要介质））的自发性血管超敏反应，以及以血清副蛋白存在为特征的B细胞肿瘤。SJL小鼠中控制对HA的全身血管超敏反应（Shs）的隐性基因座被定位于小鼠6号染色体上与人类3p25.3共线的区间，该区间与人类SCLS具有主要的遗传关联。该区间包含几个候选基因，将在SCLS队列中进行分析。