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 这项资助申请的重点是毛细血管畸形(CM)，这是一种散发的、非遗传性的血管畸形 影响到1/300的新生儿。CMS在出生时就存在，可能会影响皮肤的任何区域。它们变得更暗了， 随着时间的推移会变得更厚。病变含有过多、增大的毛细血管，并导致软组织和 骨骼过度生长。患者因皮损的出现而遭受严重的心理社会障碍 相关的过度生长会导致出血和功能障碍。斯特奇-韦伯综合征(SWS)影响1 在20,000到50,000人中，其特征是面部CM延伸到大脑和/或 眼睛。系统性红斑狼疮患者可能会出现神经功能障碍、癫痫、青光眼和失明。Cm是 由GNAQ(p.R183Q)中富含于内皮细胞(EC)的体细胞激活突变引起。 GNAQ编码GαQ，它是异源三聚体GQ蛋白的α亚基，可激活磷脂酶Cβ。这个 GαQ的过度激活导致血管生成素2的表达显著增加。不存在针对CM和 治疗包括脉冲染料激光淡化其颜色和手术切除。SWS中的癫痫发作是 由抗癫痫药物控制。迫切需要药物治疗来防止CM的进展和 传统治疗后复发。完成这些研究将是迈向这一目标的重要步骤。 AIM 1将使用敲入ANGPT2的GFP为CM/SWS的EC功能障碍创建一种基于细胞的检测方法 轨迹作为一种容易检测的读数。我们将使用这个细胞系统进行FDA的高通量筛选- 批准的药物和生物活性化合物。这可能导致识别出可用药途径。我们的 对CM形成和生长的理解，以及我们测试潜在药物治疗的能力，都受到了阻碍 因为没有老鼠模型。AIM 2将专注于在小鼠身上创造CMS。我们已经产生了一条老鼠线 其中G-αQ-R183Q在ECs中的表达可以用CdH_5Creer激活。打开GαQ-R183Q 表达的方式产生的CMS类似于人类的情况，我们将使用他莫昔芬外用。我们 将测试不同剂量的他莫昔芬和时间点(从新生儿到成年人)。我们还将通过以下方式诱导CM的形成 将腺病毒-CDH5Cre注射到出生前和出生后动物的四肢以及小鼠的脑内 ROSA-GT-GNAQ-R183Q动物获得SWS表型。动物模型的创建将使未来 从目标1测试候选药物阻止CMS形成和生长的能力的研究。最多的 有效的药物将被转化到人类身上，并进行临床试验。