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集中于毛细血管畸形（CM）（以前称为“港口染色”），是最多的常见的血管畸形类型。 CM，超出表面下方的超级毛细管样血管皮肤的是零星的先天性病变，形成结节并引起软组织和骨骼在污渍下过度生长。 Sturge-Weber综合征（SWS）是一种与脸部的CM，瘦素和眼睛的脉络膜；患者患有神经系统缺陷，青光眼。重要的是，CMS的药物治疗不存在，也无法治愈。 2013年在非综合皮肤CMS中发现了GNAQ（P.R183Q）中的体细胞激活突变 SWS CM为这项理解的血管畸形的分子研究奠定了基础。 GNAQ编码 GαQ，激活磷脂酶Cβ的异三聚体GQ蛋白的α-亚基。我们证明了 GNAQ R183Q等位基因富集在皮肤CM和SWS脑的内皮细胞（EC）中标本。我们一直在创建细胞和小鼠模型，以阐明GNAQ突变如何影响EC功能，这些变化如何导致广告以及我们如何防止广告的形成或增长。我们表明，具有R183Q突变的人类EC对层状剪切应力没有正确的反应，无法形成一个内皮屏障，并植入小鼠时形成增强的CM样血管。我们暗示蛋白质激酶C（PKC）和Angiopoietin-2（Angpt2）作为逆转GNAQ R183Q驱动的商业的潜在目标。我们正在朝着小鼠的GNAQ R183Q进行诱导，内皮特异性敲击方面的进步我们发现了在他莫昔芬诱导的敲入突变等位基因表达上的CM样病变。在此提案中，我们将确定带有体细胞GNAQ R183Q等位基因的细胞类型的广度以及如何突变改变了相同表型的转录谱与非突变细胞（AIM 1）。我们将在小鼠和斑马鱼中开发新型动物模型，以阐明导致CM的细胞步骤，并将使用它们是测试候选药物的平台（AIM 2）。我们将深入询问（Angpt2）作为一个组成性活性，突变GαQ的下游功能介质（AIM 3）。这些研究将加深我们了解GαQ活性参与者如何参与毛细血管形态发生，从而导致了第一个动物模型 CM/SWS，并提供一个可以预防或回归CM的药物的平台。关于 CM的病理生理学还将帮助我们了解其他血管病变的机制并提高我们确定防止血管过度生长（例如癌症）和在组织修复或工程期间促进血管生长。

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