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Capillary malformation: From somatic GNAQ mutations to disrupted endothelial biology

毛细血管畸形：从体细胞 GNAQ 突变到内皮生物学破坏

基本信息

批准号：
10630310
负责人：
Joyce E. Bischoff
金额：
$ 84.07万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10630310
关键词：
Affect Alleles Angiopoietin-2 Animal Model Automobile Driving Bar Codes Biology Blood Vessels Blood capillaries Brain Cell Separation Cell model Cell physiology Cells Choroid Cognitive Collaborations Coupled Cutaneous Data Defect Dermis Development Endothelial Cells Endothelium Event Experimental Models Eye Face Functional disorder G alpha q Protein G-substrate GNAQ gene Gene Expression Profile Genes Genomics Genotype Glaucoma Goals Growth Hematopoietic Hemorrhage Heterogeneity Heterotrimeric G Protein Subunit Histologic Human Image Impairment Implant In Situ Hybridization In Vitro Infant Inflammatory Knock-in Lead Leptomeninges Lesion Malignant Neoplasms Massachusetts Mediator Medical Medical center Modeling Molecular Morphogenesis Mus Mutation NF-kappa B Neurocutaneous Syndromes Neurologic Nodule Nude Mice Pathogenesis Pathology Pathway interactions Patients Pharmaceutical Preparations Pharmacotherapy Phenotype Phospholipase Play Port-Wine Stain Protein Kinase C Reporting Resistance Retinal Detachment Role Seizures Signal Transduction Skin Small Interfering RNA Somatic Mutation Sorting Specimen Stains Sturge-Weber Syndrome Surface TNFSF5 gene Tamoxifen Technology Testing Time Tissue Engineering Transcript Universities Work Zebrafish causal variant cell type cerebral capillary drug candidate drug testing high risk improved in vivo insight knock-down malformation mouse model mutant nervous system disorder novel prevent response shear stress single-cell RNA sequencing skeletal small hairpin RNA small molecule inhibitor soft tissue stroke-like episode subcutaneous tissue repair transcriptome sequencing venule

项目摘要

Project Abstract Our studies are focused on capillary malformation (CM) (previously referred to as “port-wine stain”), the most common type of vascular malformation. CM, excessive, enlarged capillary-like vessels just below the surface of the skin, are sporadic congenital lesions that darken, form nodules, and cause soft-tissue and skeletal overgrowth beneath the stain. Sturge-Weber syndrome (SWS) is a neurocutaneous disorder associated with CMs of the face, leptomeninges, and the choroid of the eye; patients suffer from neurological defects and glaucoma. Importantly, drug treatment for CMs does not exist and there is no cure. The 2013 discovery of a somatic activating mutation in GNAQ (p.R183Q) in non-syndromic cutaneous CMs and SWS CMs set the stage for molecular studies of this understudied vascular malformation. GNAQ encodes Gαq, the α-subunit of the heterotrimeric Gq protein that activates phospholipase Cβ. We showed that the GNAQ R183Q allele is enriched in the endothelial cell (EC) sorted from cutaneous CM and SWS brain specimens. We have worked on creating cellular and mouse models to elucidate how the GNAQ mutation affects EC function, how these alterations lead to CM, and how we can prevent the formation or growth of CM. We show that human ECs with the R183Q mutation do not respond properly to laminar shear stress, fail to form an endothelial barrier, and form enlarged CM-like vessels when implanted into mice. We implicate protein kinase C (PKC) and angiopoietin-2 (ANGPT2) as potential targets to reverse the GNAQ R183Q-driven CM. We are making strong progress towards an inducible, endothelial-specific knock-in of Gnaq R183Q in mice in which we have found CM-like lesions upon tamoxifen-induced expression of the knocked-in mutant allele. In this proposal we will identify the breadth of cell types that carry the somatic GNAQ R183Q allele and how the mutation alters the transcriptional profile versus non-mutant cells of the same phenotype (Aim 1). We will develop novel animal models in mice and zebrafish to elucidate the cellular steps leading to CM and will use them as platforms for testing candidate drugs (Aim 2). We will deeply interrogate the role of (ANGPT2) as a downstream functional mediator of constitutively active, mutant Gαq (Aim 3). These studies will deepen our understanding of how Gαq activity participates in capillary morphogenesis, result in the first animal models for CM/SWS, and provide a platform to test drugs that can prevent or regress CM. Discoveries about the pathophysiology of CM will also help us understand the mechanisms that underlie additional vascular lesions and improve our ability to identify new pathways for preventing vascular overgrowth (e.g., cancer) and promoting vascular growth during tissue repair or engineering.

项目摘要我们的研究主要集中在毛细血管畸形（CM）（以前称为"葡萄酒斑"），血管畸形的常见类型。CM，表面下过度、扩大的毛细血管样血管是散发性的先天性病变，会变暗，形成结节，并导致软组织和骨骼污渍下的过度生长Sturge-Weber综合征（SWS）是一种神经皮肤疾病，面部、软脑膜和眼脉络膜的CM;患者患有神经缺陷，青光眼重要的是，CM的药物治疗不存在，也没有治愈方法。 2013年在非综合征性皮肤CM中发现GNAQ（p.R183Q）的体细胞激活突变 SWS CM为这种未充分研究的血管畸形的分子研究奠定了基础。GNAQ编码 G α q，异源三聚体Gq蛋白的α亚基，可激活磷脂酶C β。我们证明了 GNAQ R183Q等位基因在从皮肤CM和SWS脑中分选的内皮细胞（EC）中富集标本我们一直致力于创建细胞和小鼠模型，以阐明GNAQ突变是如何影响EC功能，这些改变如何导致CM，以及我们如何预防CM的形成或生长。我们发现，具有R183Q突变的人内皮细胞对层流剪切应力没有适当的反应，形成内皮屏障，并在植入小鼠体内时形成扩大的CM样血管。我们认为蛋白质激酶C（PKC）和血管生成素-2（ANGPT 2）作为逆转GNAQ R183Q驱动的CM的潜在靶点。我们在小鼠中的Gnaq R183Q的诱导性、内皮特异性敲入方面取得了很大进展，我们已经发现在他莫昔芬诱导的敲入突变等位基因表达后出现CM样病变。在这个建议中，我们将确定携带体细胞GNAQ R183Q等位基因的细胞类型的广度，以及如何与相同表型的非突变细胞相比，突变改变了转录谱（Aim 1）。我们将在小鼠和斑马鱼中开发新的动物模型，以阐明导致CM的细胞步骤，并将使用作为测试候选药物的平台（目标2）。我们将深入询问（ANGPT2）作为一个组成型活性突变体G α q的下游功能性介质（Aim 3）。这些研究将加深我们的了解G α q活性如何参与毛细血管形态发生，导致第一个动物模型， CM/SWS，并提供一个平台来测试可以预防或消退CM的药物。发现关于 CM的病理生理学也将帮助我们理解其他血管病变的机制并提高我们识别防止血管过度生长的新途径的能力（例如，癌症）和在组织修复或工程中促进血管生长。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A somatic missense mutation in GNAQ causes capillary malformation.

DOI：
10.1097/moh.0000000000000500
发表时间：
2019-05
期刊：
Current opinion in hematology
影响因子：
3.2
作者：
Bichsel C;Bischoff J
通讯作者：
Bischoff J

MRC1 and LYVE1 expressing macrophages in vascular beds of GNAQ p.R183Q driven capillary malformations in Sturge Weber syndrome.

Sturge Weber 综合征中 GNAQ p.R183Q 血管床中表达 MRC1 和 LYVE1 的巨噬细胞导致毛细血管畸形。

DOI：
10.1186/s40478-024-01757-4
发表时间：
2024
期刊：
Acta neuropathologica communications
影响因子：
7.1
作者：
Nasim,Sana;Bichsel,Colette;Dayneka,Stephen;Mannix,Robert;Holm,Annegret;Vivero,Mathew;Alexandrescu,Sanda;Pinto,Anna;Greene,ArinK;Ingber,DonaldE;Bischoff,Joyce
通讯作者：
Bischoff,Joyce