Studies in the Pathogenesis of Systemic Capillary Leak Syndrome

免疫失调可能导致scs的病理生理表现。MGUS是多发性骨髓瘤（MM）的恶性前体细胞，是一种以分泌单克隆免疫球蛋白（Ig，也称为副蛋白）的克隆浆细胞群为特征的疾病。在大多数scs病例中可检测到MGUS。一些MGUS演变为骨髓瘤或浆细胞白血病的scs患者在造血功能障碍化疗后毛细血管渗漏事件较少。这些发现表明，来自失调浆细胞群的单克隆副蛋白可能是观察到的病理生理结果的直接或间接来源。目前的研究主要集中在使用高通量蛋白质阵列识别SCLS单克隆Ig的靶抗原（如果有的话）。

Immune dysregulation may contribute to the pathophysiologic findings seen in SCLS. A MGUS, a premalignant precursor to multiple myeloma (MM), is a condition characterized by a clonal plasma cell population that secretes monoclonal immunoglobulins (Ig, also referred to as a paraprotein). MGUS is detectable in a majority of SCLS cases. Several patients with SCLS in whom MGUS evolved into 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. Current studies are focused on identifying the target antigen of SCLS monoclonal Ig, if any, using high-throughput protein arrays. 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 SCLS symptoms and/or etiological factors can be identified. We have now evaluated more than 75 patients with a confirmed diagnosis of SCLS under this protocol in the last 12 years. We are the primary referral center in the U.S. for SCLS. Circulating permeability factors, vascular endothelial growth factor (VEGF), angiopoietin 2 (Angpt-2), CXCL10, CCL2, and IL-6, were elevated in episodic SCLS sera compared to remission sera. These findings suggest that angiogenic proteins and proinflammatory cytokines that induce endothelial cell (EC) hyper-permeability may contribute to transient EC barrier dysfunction around SCLS flares. The transient episodes of hypotensive shock and anasarca in SCLS are thought to arise from reversible microvascular barrier dysfunction. Application of episodic but not convalescent SCLS sera to human microvascular ECs caused vascular endothelial cadherin internalization, disruption of interendothelial junctions, actin stress fiber formation, and increased permeability in complementary functional assays. EC contraction and temporary attenuation of adherens junctions may thus permit leakage of solutes and proteins into the extravascular space during acute episodes. In fiscal year 21, we continued to study mechanisms underlying the discovery that the skin microvasculature and endothelial cell lines from SCLS patients are hyper-responsive to routine inflammatory mediators such as VEGF and histamine. Preliminary data suggest that SCLS cells have a primary metabolic abnormality that renders them more susceptible to these mediators. The role of specific gene defects in SCLS, if any, is unknown; e.g. whether the endothelium is genetically programmed for hyper-responsiveness to routine stimuli. There are no consistent familial aggregations in SCLS, and whole exome sequencing of unrelated adults with SCLS and several parent-child trios failed to reveal a uniform exotic etiology. 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. From unbiased high-density mapping of single-nucleotide polymorphisms (SNPs), a small genetic interval, 3p25.3, was identified as the highest-ranking candidate susceptibility locus (p 10-6) with an odds ratio of 41. Odds ratios (7-41) and p values (10-4 and 10-6) for the top SCLS-associated variants were outsized for such a small sample size. These results imply high penetrance for a rare disease allele that remains to be identified. Currently, we are performing whole genome sequencing of DNA from patients to test the hypothesis that they are prone to exaggerated responses to otherwise mundane inflammatory stressors due to underlying genetic defect(s). In FY22, we reported a case series describing the treatment of acute SCLS episodes with high dose intravenous immunoglobulins (IVIG). In this retrospective study, administration of IVIG with minimal additional IV fluids was safe and associated with rapid clinical improvement. We concluded that IVIG given close to the onset of ISCLS-related symptoms is associated with a favorable outcome. Secondly, in FY22, we reported the use of exome sequencing for the study of rare diseases of unknown etiology including SCLS. We participated in a program that integrates exome sequencing, chromosomal microarray, phenotyping, results return with genetic counseling, and reanalysis in 1505 individuals from 1000 families with suspected or known inborn errors of immunity. The study demonstrated that comprehensive analysis of exome data has diagnostic and clinical utility for patients with suspected immune disorders. Finally, we pursued a collaboration with PharmAbcine, Inc. to study a monoclonal antibody, PMC-403, which targets the Angpt1/2 receptor, Tie2. PMC-403 is a novel agonistic antibody that binds to Tie2 receptor and promotes stabilization of leaky blood vessels in tumor models in mice. The key findings were presented recently at the American Association of Cancer Research (AACR) meeting 2020, and PMC-403 is expected to enter a phase I clinical trial in 2022. We demonstrated that PMC-403 ameliorates vascular leakage in preclinical mouse models of SCLS and in SCLS-patient derived ECs challenged with inflammatory mediators in vitro.