Drug Development for Pediatric Capillary Malformation

治疗小儿毛细血管畸形的药物开发

• 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
• 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

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.

项目摘要 这项拨款申请的重点是毛细血管畸形（CM），一种散发性，非遗传性血管异常 影响1/300的新生儿CM在出生时就存在，可能会影响皮肤的任何区域。它们变得更黑， 随着时间的推移越来越厚。病变包含过多的，扩大的毛细血管样血管，并导致软组织和 骨骼过度生长。患者因病变的出现而遭受严重的心理社会疾病， 相关的过度生长可导致出血和功能性残疾。Sturge-Weber综合征（SWS）累及1例 在20，000至50，000人中，其特征在于面部CM延伸到大脑和/或 眼睛SWS患者可能会出现神经功能障碍、癫痫发作、青光眼和失明。CM是 由内皮细胞（EC）中富集的GNAQ（p.R183Q）中的体细胞激活突变引起。 GNAQ编码Gαq，其为激活磷脂酶Cβ的异源三聚体Gq蛋白的α亚基。的 Gαq的过度激活导致ANGPT 2表达的强烈增加。CM不存在药物， 治疗包括脉冲染料激光以减轻其颜色和手术切除。SWS的缉获量为 由抗癫痫药物控制。迫切需要药物治疗来预防CM进展， 传统治疗后复发。完成这些研究将是实现这一目标的重要步骤。 目的1将使用敲入ANGPT 2的GFP建立CM/SWS中EC功能障碍的基于细胞的测定 作为一个容易检测的读数。我们将使用这种细胞系统进行FDA的高通量筛选- 批准的药物和生物活性化合物。这可能会导致确定药物途径。我们 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， 因为缺乏小鼠模型。目标2将专注于在小鼠中创建CM。我们已经生成了一条鼠标线 其中我们可以使用Cdh 5CreER激活EC中Gα q-R183 Q表达。开启Gα q-R183 Q 为了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）。我们还将诱导CM形成， 将腺病毒-Cdh 5Cre注射到产前和产后动物的四肢中，以及注射到 R 0 SA-GT-GNAQ-R183 Q动物以获得SWS表型。动物模型的创建将使未来成为可能 研究测试目标1中的候选药物阻止CM形成和生长的能力。最 有效的药物将转化为人类并进行临床试验。