Vascular Pathogenesis of Port Wine Stain

波特酒色斑的血管发病机制

• 批准号: 10378523
• 负责人: Wenbin Tan
• 金额: $ 30.58万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-08 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10378523
• 关键词: Area; Basement membrane; Biological Markers; Biopsy; Blood Circulation; Blood Vessels; Cardiovascular system; Cell Proliferation; Cell model; Cells; Child; Congenital Abnormality; Cutaneous; Data; Dermal; Development; Dilatation - action; Disintegrins; Dyes; Endothelial Cells; Endothelium; Etiology; Excision; Exocytosis; Extracellular Matrix; Genes; Goals; Hemangioma; Human; In Vitro; Incidence; Individual; Infant; KDR gene; Knowledge; Lasers; Lesion; Live Birth; Mediator of activation protein; Metalloproteases; Molecular; Molecular Profiling; Molecular Target; Names; National Institute of Arthritis and Musculoskeletal and Skin Diseases; Nude Mice; PECAM1 gene; Pathogenesis; Pathologic; Pathway interactions; Patients; Pericytes; Permeability; Persons; Phenotype; Phosphotransferases; Physiologic pulse; Play; Port-Wine Stain; Prevalence; Protein Analysis; Proteins; Research; Role; Serum; Severities; Signal Transduction; Site; Skin; Testing; United States; Venous; Xenograft Model; Xenograft procedure; angiogenesis; clinically significant; effective therapy; exosome; in vivo; in vivo Model; malformation; migration; novel; novel therapeutic intervention; therapeutically effective; therapy outcome; transcriptome; treatment choice; venule

PROJECT SUMMARY Port Wine Stain (PWS) is a congenital, progressive vascular malformation of human skin which occurs in an estimated incidence of 3-5 infants per 1,000 live births. Approximately 1.2 million individuals in the United States have PWS birthmarks. Pulsed dye laser (PDL) remains the choice of treatment for PWS. However, only less than 10% of patients achieve complete lesion fading after PDL. Inadequate PWS therapeutic outcome is a clinically significant problem that requires an urgent solution. The etiology and pathogenesis of PWS are currently poorly understood. This knowledge gap is a major obstacle for developing any new effective treatments. PWS is one of the research areas under the NIAMS long-range plan listed as “understanding the cause of skin vasculature birth defects (e.g., hemangioma and port wine stain) and developing effective therapies.” The studies proposed herein are specifically directed towards understanding pathological mechanisms of PWS and facilitating the development of new, more effective approaches to treatment. In searching for causative substances for PWS, we have identified that circulating CD31+ exosomes, which are derived from human lesional dermal microvascular endothelial cells (hDMVECs), facilitate the formation of vascular phenotypes of PWS. Analysis of the protein content of the serum CD31+ exosomes reveals that PWS CD31+ exosomes harbor a unique molecular milieu that include EphB1, ephrinB2 (EfnB2), a-disintegrin-and- metalloprotease domain 30 (ADAM30), and exocytosis mediator synatotagamin like-1 (SYTL1). Functional analysis shows ADAM30 functions as a sheddase to cleave EfnB2 and its activity is regulated by vascular endothelial growth factor receptor 2 (VEGFR2) signaling. More interestingly, compared to normal controls, PWS serum CD31+ exosomes and hDMVEC culture-derived EphB1/EfnB2/ADAM30 exosomes possess much greater angiogenesis activity. They greatly increased the number and size of new blood vessels formed in a nude mouse xenograft model in vivo. These novel findings motivate us to test a hypothesis that hDMVEC- derived CD31+ exosomes enrich at lesional sites and cause the development and progression of PWS via disruption of cell-cell and cell-ECM interactions and EC barrier function through a molecular pathway involving exosomal EphB1/EfnB2/ADAM30 (Fig. 1). These exosomes are also released into the circulatory system where they can be identified and serve as potential serum biomarkers. In Aim 1, we will determine the specific molecular profiles of CD31+ exosomes derived from PWS ECs as compared to normal controls in vitro and in vivo. We will determine if secretion of CD31+ exosomes from PWS hDMVECs is regulated by exocytosis genes SYTL1 and synatotagamin-1 (SYT1) in vivo and in vitro; In Aim 2, we will determine the mechanisms by which EphB1/EfnB2 regulates VEGFR2 signaling to activate ADAM30 leading to subsequent EfnB2 cleavage; in Aim 3, we will determine if PWS CD31+ exosomal EphB1/EfnB2/ADAM30 induces the formation of PWS vascular phenotypes in vitro; and in Aim 4, we will determine if PWS CD31+ exosomal EphB1/EfnB2/ADAM30 induces angiogenesis and formation of PWS vascular phenotypes in vivo. The long- term goal of this research is to investigate the pathogenesis of PWS for developing new treatments of PWS. The immediate goal is to establish the role of lesional endothelial exosomes in the vascular phenotypes in the pathogenesis of PWS. Our proposed studies are totally novel and data so obtained will be important since they will define a novel molecular pathway for the pathogenesis of PWS and open new avenues for the development of more effective therapeutic strategies for PWS.

项目摘要 鲜红斑痣（PWS）是一种先天性、进行性的人类皮肤血管畸形， 估计发病率为每1 000名活产婴儿中有3-5名婴儿。美国约有120万人 美国有PWS胎记。脉冲染料激光（PDL）仍然是治疗PWS的选择。但只有 少于10%的患者在PDL后实现完全病变消退。PWS治疗结局不充分是一种 需要紧急解决的临床重大问题。PWS的病因和发病机制是 目前了解甚少。这种知识差距是开发任何新的有效的 治疗。PWS是NIAMS长期计划下的研究领域之一，被列为“了解 皮肤脉管系统出生缺陷的原因（例如，血管瘤和鲜红斑痣）和开发有效的 治疗”。本文提出的研究是专门针对了解病理 PWS的机制，并促进新的，更有效的治疗方法的发展。在 为了寻找PWS的致病物质，我们已经确定了循环CD 31+外泌体， 来源于人皮损真皮微血管内皮细胞（hDMVEC），促进 PWS的血管表型。对血清CD 31+外泌体的蛋白质含量的分析表明，PWS CD 31+外泌体具有独特的分子环境，包括EphB 1、肝配蛋白B2（EfnB 2）、α-去整合素和β-去整合素。 金属蛋白酶结构域30（ADAM 30）和胞吐介质突触素样蛋白-1（SYTL 1）。功能 分析表明，ADAM 30作为脱落酶切割EfnB 2，其活性受到血管内皮细胞的调节。 内皮生长因子受体2（VEGFR 2）信号传导。更有趣的是，与正常对照组相比， PWS血清CD 31+外泌体和hDMVEC培养物衍生的EphB 1/EfnB 2/ADAM 30外泌体具有许多 更强的血管生成活性。他们大大增加了新生血管的数量和大小， 裸鼠体内异种移植模型。这些新的发现促使我们测试一个假设，即hDMVEC- 衍生的CD 31+外泌体在病变部位富集，并通过以下途径引起PWS的发展和进展： 通过分子途径破坏细胞-细胞和细胞-ECM相互作用以及EC屏障功能 涉及外泌体EphB 1/EfnB 2/ADAM 30（图1）。这些外泌体也被释放到循环系统中 系统中，它们可以被识别并作为潜在的血清生物标志物。在目标1中，我们将确定 与正常对照相比，来自PWS EC的CD 31+外泌体的特异性分子谱， 体外和体内。我们将确定来自PWS hDMVEC的CD 31+外泌体的分泌是否受到以下因素的调节： 胞吐基因SYTL 1和synatotagamin-1（SYT 1）在体内和体外;在目的2中，我们将确定 EphB 1/EfnB 2调节VEGFR 2信号传导以激活ADAM 30的机制，导致随后的 EfnB 2切割;在目标3中，我们将确定PWS CD 31+外泌体EphB 1/EfnB 2/ADAM 30是否诱导EfnB 2切割。 在目标4中，我们将确定PWS CD 31+外泌体是否与PWS CD 31+外泌体融合。 EphB 1/EfnB 2/ADAM 30在体内诱导血管生成和PWS血管表型的形成。很长的- 本研究的长期目标是探讨PWS的发病机制，以开发PWS的新治疗方法。 当前的目标是确定病变内皮外泌体在血管表型中的作用， PWS的发病机制。我们提出的研究是完全新颖的，因此获得的数据将是重要的，因为它们 将为PWS的发病机制定义一个新的分子途径，并为PWS的治疗开辟新的途径。 开发更有效的PWS治疗策略。

项目成果

Reprogramming endothelial and vascular smooth muscle cells to prevent and treat hypertension.

• DOI: 10.1016/j.mehy.2023.111162
• 发表时间: 2023-09
• 期刊: Medical hypotheses
• 影响因子: 4.7
• 作者: Laena Pernomian;W. Tan;Cameron G. McCarthy;C. F. Wenceslau

Endothelial cells differentiated from patient dermal fibroblast-derived induced pluripotent stem cells resemble vascular malformations of port-wine birthmark.

由患者真皮成纤维细胞衍生的诱导多能干细胞分化而来的内皮细胞类似于葡萄酒胎记的血管畸形。

• DOI: 10.1093/bjd/ljad330
• 发表时间: 2023
• 期刊: The British journal of dermatology
• 作者: Nguyen,Vi;Gao,Chao;Hochman,MarceloL;Kravitz,Jacob;Chen,ElliottH;Friedman,HaroldI;Wenceslau,CamillaF;Chen,Dongbao;Wang,Yunguan;Nelson,JStuart;Jegga,AnilG;Tan,Wenbin
• 通讯作者: Tan,Wenbin

The Spike Protein of SARS-CoV-2 Impairs Lipid Metabolism and Increases Susceptibility to Lipotoxicity: Implication for a Role of Nrf2.

• DOI: 10.3390/cells11121916
• 发表时间: 2022-06-14
• 期刊: Cells
• 影响因子: 6

Membrane trafficking and exocytosis are upregulated in port wine stain blood vessels.

• DOI: 10.14670/hh-18-051
• 发表时间: 2019-05
• 期刊: Histology and histopathology
• 影响因子: 2
• 作者: Yin R;Rice SJ;Wang J;Gao L;Tsai J;Anvari RT;Zhou F;Liu X;Wang G;Tang Y;Mihm MC Jr;Belani CP;Chen DB;Nelson JS;Tan W
• 通讯作者: Tan W

Roles of Exosomes in Cardiac Fibroblast Activation and Fibrosis.

• DOI: 10.3390/cells10112933
• 发表时间: 2021-10-28
• 期刊: Cells
• 影响因子: 6
• 作者: Hohn J;Tan W;Carver A;Barrett H;Carver W
• 通讯作者: Carver W