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 该赠款申请的重点是毛细血管畸形（CM），这是一种零星的非遗传性血管异常 影响1/300名新生儿。 CM出生时存在，可能会影响皮肤的任何区域。他们变得黑暗 随着时间的流逝，更厚。病变中包含过多的，毛细管样的血管，并引起软组织，并 骨骼过度生长。患者因病变的出现而遭受严重的社会心理发病率 相关的过度生长会导致出血和功能障碍。 Sturge-Weber综合征（SWS）影响1 在20,000至50,000个人中，其特征是面部CM，向大脑和/或 眼睛。 SWS患者可能会出现神经系统障碍，癫痫发作，青光眼和失明。 CM是 由富含内皮细胞（EC）的GNAQ（P.R183Q）中的体细胞激活突变引起。 GNAQ编码激活磷脂酶Cβ的异三聚体GQ蛋白的α-亚基。这 GαQ的过度激活导致ANGPT2表达的强烈增加。 CM不存在药物，并且 管理由脉冲型激光器组成，以减轻其颜色和外科手术的去除。 SWS中的癫痫发作是 由抗癫痫药控制。迫切需要药物治疗以防止CM进展和 传统治疗后的复发。这些研究的完成将是实现这一目标的主要步骤。 AIM 1将使用GFP撞到ANGPT2的GFP来创建基于细胞的EC功能障碍的EC功能障碍 座位作为易于检测的读数。我们将使用此细胞系统进行FDA-的高通量筛选 批准的药物和生物活性化合物。这可能导致识别可吸毒途径。我们的 了解CM形成和生长以及我们测试潜在药物治疗的能力的理解受到阻碍 由于没有鼠标模型。 AIM 2将专注于在小鼠中创建CMS。我们已经生成了一条鼠标 其中我们可以使用CDH5CREER激活EC中GαQ-R183Q的表达。打开GαQ-R183Q 以产生重新组装人类状况的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（成人新生）。我们还将通过 将腺病毒-CDH5CRE注入产前和产后动物的四肢，以及 ROSA-GT-GNQ-R183Q动物获得SWS表型。动物模型的创建将使未来 从AIM 1测试候选药物的研究，以阻止CMS的形成和生长。最多 简单的药物将转化为人类，并接受临床试验。