Understanding and targeting molecular and cellular events responsible for pulmonary arteriovenous malformation development, growth and regression

了解和靶向导致肺动静脉畸形发生、生长和消退的分子和细胞事件

• 批准号: 10718086
• 负责人: Edda Frauke Spiekerkoetter
• 金额: $ 69.62万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10718086
• 关键词: Ablation; Abnormal Endothelial Cell; Adult; Animal Model; Apoptosis; Arteries; Arteriovenous malformation; BMPR2 gene; Blood capillaries; Brain; Bypass; CRISPR correction; Catheters; Cell physiology; Cells; Cerebral Abscesses; Child; Childhood; Clinical Trials; Color; Conceptions; Data; Development; Disease; Disease model; Dominant Genetic Conditions; Endoglin; Endothelial Cells; Endothelium; Epistaxis; Event; FK506; Fluorescent in Situ Hybridization; Functional disorder; Gastrointestinal tract structure; Genes; Genetic Diseases; Growth; Growth and Development function; Health; Hemorrhage; Hereditary hemorrhagic telangiectasia; Heterozygote; Human; Image; In Vitro; Intervention; Knowledge; Label; Life; Liver; Loss of Heterozygosity; Lung; MADH4 gene; Maps; Mediating; Medical; Migraine; Mission; Molecular; Molecular Target; Morbidity - disease rate; Mus; Mutation; Organ; Organ Culture Techniques; Pathologic; Pathway interactions; Patients; Penetrance; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phenotype; Physiologic pulse; Population; Preclinical Testing; Prevalence; Prevention; Proliferating; Radiation exposure; Reperfusion Therapy; Research; Resolution; Risk; Role; Signal Pathway; Signal Transduction; Skin; Slice; Small Interfering RNA; Stroke; Structure of mucous membrane of nose; Tacrolimus; Techniques; Testing; Therapeutic; Therapeutic Embolization; Three-Dimensional Imaging; Tissues; Tube; United States National Institutes of Health; Vascular Endothelial Cell; Veins; Venous; Visceral; autosome; capillary bed; cell behavior; cell type; clinically relevant; common symptom; disability; drug candidate; drug development; drug repurposing; high throughput screening; high-throughput drug screening; improved; in silico; in vivo; induced pluripotent stem cell; innovation; knock-down; lead candidate; loss of function mutation; lung imaging; lung microvascular endothelial cells; mortality; mortality risk; mouse model; novel; novel strategies; novel therapeutic intervention; novel therapeutics; pharmacologic; postnatal; prevent; pulmonary arterial hypertension; single molecule; therapeutic evaluation; therapeutic target; tissue culture; tool; transcriptome sequencing; transcriptomics; vascular bed

Hereditary hemorrhagic telangiectasia (HHT) is a genetic disease characterized by multiple arteriovenous malformations (AVMs) which are direct connections between arteries and veins, bypassing the capillary bed. Pulmonary AVMs (PAVMs) are the most common visceral AVMs in adult (10-45%) and pediatric HHT patients (60%) and cause significant morbidity and mortality due to an increased risk for cerebral abscesses, stroke, pulmonary hemorrhage and migraines. Current treatment for PAVMs consists of catheter mediated embolization with a re-perfusion rate of up to 25%, necessitating frequent imaging (radiation exposure) as well as repeat interventions. While heterozygous loss-of function mutations in ENDOGLIN, ALK1 and SMAD4 are responsible for the development of HHT in 85% of patients, we still do not know precisely how PAVMs develop. In particular, we do not know exactly from which vascular bed (arterial, capillary, venous) PAVMs arise, and which downstream signaling pathway is most important for PAVM development or growth that could be harnessed as a therapeutic target. No medical therapy exists that is able to prevent, arrest growth or even reverse PAVMs. Furthermore, we are lacking precise animal models of PAVMs or in vitro disease models, necessary for pre-clinical testing of therapeutic approaches. We therefore hypothesized that understanding the cellular and molecular mechanisms governing PAVM development paired with the identification of clinically relevant, pathological signaling abnormalities will allow us to develop and test novel therapeutic approaches that prevent and potentially reverse disease. Our proposal has three significant parts, which are represented by our three specific aims: First, to develop and characterize a novel mouse model of PAVM formation by deleting HHT causing genes in different endothelial cell subpopulations and study their role in PAVM development and growth. Second, to differentiate induced pluripotent stem cells (iPSCs) from HHT patients into arterial and venous endothelial cells (ECs), to identify novel common or unique pathways altered in HHT as a direct consequence of mutations in ENG, ALK1 and SMAD4, to predict repurposed drugs (in silico) and test whether they target the newly identified pathways in iPSC-ECs and tissue culture. Third, to test whether lead candidate drugs, FK506 and Enzastaurin, and novel drugs identified in Aim 2 (ie Brivanib, see preliminary data) positively influence PAVM formation, growth and potential regression. Our proposal is innovative because it combines a conceptionally novel approach (understanding PAVM development by focusing on disease-causing alterations in subpopulations of lung endothelial cells) with cutting edge techniques (multiplex single-molecule fluorescence in situ hybridization, spatial transcriptomics, multicolor labeling and high resolution 3-D imaging of the lung) and novel pharmacological interventions (drugs identified by High-Throughput Screening, predicting novel drugs in silico).The short-term impact will be a better understanding of how AVMs form in the lung and potentially in other organs (brain, skin). The long-term impact will be the identification of potential novel treatments for AVMs.

遗传性出血性毛细血管扩张症是一种以多发性动静脉为特征的遗传性疾病。 动静脉畸形(AVM)是动脉和静脉之间绕过毛细血管床的直接连接。 肺动静脉畸形(PAVM)是成人(10%-45%)和儿童HHT患者最常见的内脏AVM (60%)，并导致显著的发病率和死亡率，因为脑脓肿、中风、 肺出血和偏头痛。目前对肺静脉畸形的治疗包括导管介入性栓塞 再灌注率高达25%，需要频繁成像(辐射照射)以及重复 干预措施。虽然endoglin、ALK1和Smad4的杂合性功能丧失突变是原因 对于85%的患者发生HHT，我们仍然不确切地知道PAVMS是如何发生的。在……里面 具体地说，我们不确切地知道哪些血管床(动脉、毛细血管、静脉)产生了PAVM，以及哪些 下游信号通路对PAVM的发育或生长最重要，可以被利用为 一个治疗靶点。目前还没有能够预防、阻止生长甚至逆转的药物疗法。 PAVM。此外，我们缺乏精确的PAVMS动物模型或体外疾病模型，这是必要的 用于治疗方法的临床前测试。因此，我们假设，理解细胞 控制PAVM发展的分子机制与临床相关的 病理信号异常将使我们能够开发和测试新的治疗方法，以防止 并有可能逆转疾病。我们的提案有三个重要部分，它们由我们的三个 具体目标：首先，通过缺失HHT，建立和鉴定一种新的PAVM形成的小鼠模型 导致不同内皮细胞亚群中的基因，并研究它们在PAVM发育和生长中的作用。 第二，将HHT患者的诱导多能干细胞(IPSCs)分化为动脉和静脉 内皮细胞(ECs)，以确定HHT中新的常见或独特的通路作为HHT的直接结果 ENG、ALK1和Smad4的突变，以预测改变用途的药物(在硅胶中)并测试它们是否针对 新发现的IPSC-ECs和组织培养途径。第三，为了测试主要候选药物FK506 和enzastaurin，以及AIM 2中发现的新药(即brivanib，见初步数据)对PAVM有积极影响 形成、成长和潜在的回归。我们的建议是创新的，因为它结合了概念上的 新的方法(通过关注引起疾病的改变来了解PAVM的发展 肺内皮细胞亚群)和尖端技术(多重单分子荧光 原位杂交、空间转录、多色标记和高分辨率肺三维成像)和 新的药理干预措施(通过高通量筛选确定的药物，预测新药 短期影响将是更好地了解动静脉动静脉畸形是如何在肺内形成的，并可能在 其他器官(脑、皮肤)。长期影响将是确定潜在的动静脉畸形的新治疗方法。