Drug Development for Pediatric Capillary Malformation

治疗小儿毛细血管畸形的药物开发

基本信息

批准号：
10536819
负责人：
Matthew P Vivero
金额：
$ 7.17万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10536819
关键词：
ANGPT2 gene Adenoviruses Adult Affect Alleles Angiopoietin-2 Animal Model Animals Antiepileptic Agents Appearance Applications Grants Area Biological Biological Assay Birth Blindness Blood Vessels Blood capillaries Brain CRISPR/Cas technology Cells Clinical Trials Clothing Color Complementary DNA Cutaneous Deformity Diffuse Dose Dyes Eating Embryo Endothelial Cells Endothelium Enterobacteria phage P1 Cre recombinase Excision Eye FDA approved Face Fluorescence Functional disorder Future G alpha q Protein GNAQ gene Genes Genetic Glaucoma Goals Green Fluorescent Proteins Growth Hemorrhage Heterotrimeric G Protein Subunit Histologic Human Impairment In Vitro Individual Infant Injectable Injections Intervention Laboratories Laser Surgery Lasers Lead Lesion Libraries Life Limb structure Methods Modeling Molecular Morbidity - disease rate Mus Mutation Neurologic Newborn Infant Nodule Operative Surgical Procedures Oral PIK3CA gene Pain Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Pharmacotherapy Phenotype Phospholipase Physiologic pulse Port-Wine Stain Pre-Clinical Model Preclinical Testing Proteins Recurrence Safety Seizures Site Skin Sturge-Weber Syndrome System TNFSF5 gene Tamoxifen Telomerase Testing Therapeutic Uses Thick Time Translating Virus Walking Work base bone drug candidate drug testing experimental study fetal functional disability high throughput screening human disease in vivo in vivo Model malformation mouse model mutant overexpression pediatric drug development postnatal prenatal prevent promoter protein expression psychosocial skeletal small molecule soft tissue venule

项目摘要

Project Summary This grant application is focused on capillary malformation (CM), a sporadic, non-hereditary vascular anomaly affecting 1/300 newborns. CMs are present at birth and may affect any area of skin. They grow darker and thicker over time. The lesions contain excessive, enlarged capillary-like vessels and cause soft-tissue and skeletal overgrowth. Patients suffer severe psychosocial morbidity from the appearance of the lesions and associated overgrowth can cause bleeding and functional disability. Sturge-Weber syndrome (SWS) affects 1 in 20,000 to 50,000 individuals and is characterized by a facial CM with extension to either the brain and/or eyes. Patients with SWS may develop neurological impairment, seizures, glaucoma, and blindness. CM is caused by a somatic activating mutation in GNAQ (p.R183Q) that is enriched in the endothelial cell (EC). GNAQ encodes Gαq, the α- subunit of the heterotrimeric Gq protein that activates phospholipase Cβ. The overactivation of Gαq leads to a strong increase in ANGPT2 expression. Drugs do not exist for CM and management consists of pulse-dye laser to lighten its color and surgical removal. Seizures in SWS are controlled by anti-epileptic drugs. Pharmacotherapy is desperately needed to prevent CM progression and recurrence following traditional treatments. Completion of these studies will be major steps towards this goal. Aim 1 will create a cell-based assay for EC dysfunction in CM/SWS using GFP knocked into the ANGPT2 locus as an easily detectable readout. We will use this cell system for high-throughput screening of FDA- approved drugs and bioactive compounds. This could lead to the identification of druggable pathways. Our understanding of how CM forms and grows, as well as our ability to test potential drug treatments, is hampered by the absence of a mouse model. Aim 2 will focus on creating CMs in mice. We have generated a mouse line in which we can activate expression of Gαq-R183Q in ECs using Cdh5CreER. To turn on Gαq-R183Q expression in a manner that produces CMs resembling the human condition we will use topical tamoxifen. We will test different doses of tamoxifen and time points (new-born to adult). We also will induce CM formation by injection of Adenovirus-Cdh5Cre into the limbs of prenatal and postnatal animals, as well as into the brain of ROSA-GT-GNAQ-R183Q animals to obtain a SWS phenotype. Creation of an animal model will enable future studies to test drug candidates from Aim 1 for their ability to stop the formation and growth of CMs. The most efficacious drugs will be translated to humans and undergo clinical trials.

项目概要该拨款申请的重点是毛细血管畸形（CM），一种散发性的非遗传性血管异常影响 1/300 的新生儿出生时就存在 CM，并且可能影响任何皮肤区域。随着时间的推移，病变部位会变得越来越厚，毛细血管样血管增多，导致软组织和血管变厚。骨骼过度生长，患者因病变的出现而遭受严重的社会心理疾病。相关的过度生长可导致出血和功能障碍 1。在 20,000 至 50,000 个人中，其特征是面部 CM 延伸至大脑和/或 SWS 患者可能会出现神经功能障碍、癫痫、青光眼和失明。由在内皮细胞 (EC) 中富集的 GNAQ (p.R183Q) 体细胞激活突变引起。 GNAQ 编码 Gαq，即激活磷脂酶 Cβ 的异源三聚体 Gq 蛋白的 α 亚基。 Gαq 过度激活会导致 ANGPT2 表达强烈增加，但不存在针对 CM 和 CM 的药物。治疗方法包括脉冲染料激光淡化其颜色和手术切除 SWS 癫痫发作。迫切需要药物治疗来预防 CM 进展和控制。完成这些研究将是实现这一目标的重要步骤。目标 1 将使用敲入 ANGPT2 的 GFP 创建基于细胞的 CM/SWS 中 EC 功能障碍检测位点作为易于检测的读数，我们将使用该细胞系统进行 FDA-的高通量筛选。该药物可能导致我们的药物途径的鉴定。对 CM 如何形成和生长的理解以及我们测试潜在药物治疗的能力受到阻碍由于缺乏小鼠模型，目标 2 将专注于在小鼠中创建 CM。其中我们可以使用 Cdh5CreER 激活 EC 中 Gαq-R183Q 的表达来打开 Gαq-R183Q。以产生类似于人类状况的 CM 的方式表达，我们将使用局部他莫昔芬。我们将测试不同剂量的他莫昔芬和时间点（新生儿到成人）我们还将通过诱导 CM 形成。将腺病毒-Cdh5Cre注射到产前和产后动物的四肢以及大脑中 ROSA-GT-GNAQ-R183Q 动物获得 SWS 表型将使未来能够创建动物模型。研究测试目标 1 候选药物阻止 CM 形成和生长的能力。有效的药物将被转化为人体并进行临床试验。