Foxf1 Transcription Factor in Development of Pulmonary Capillaries

Foxf1转录因子在肺毛细血管发育中的作用

基本信息

批准号：
9065597
负责人：
Vladimir Kalinichenko
金额：
$ 39万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-01 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9065597
关键词：
Acute Lung Injury Alleles Alveolar Alveolar capillary dysplasia with misalignment of pulmonary veins Binding Binding Proteins Birth Blood Vessels Blood capillaries Boxing Bronchopulmonary Dysplasia Cell Proliferation Cells Congenital Disorders DNA Binding DNA-Binding Proteins Data Defect Development Disease Dominant-Negative Mutation Embryo Endothelial Cells FOXF1 gene Frameshift Mutation Functional disorder Funding Gene Expression Gene Targeting Genes Genetic Genetic Transcription Health Human Hypoxemia In Vitro Infant Infant Mortality Inherited KDR gene Knock-in Mouse Laboratories Link Lung Lung diseases Mass Spectrum Analysis Mediator of activation protein Modeling Molecular Molecular Abnormality Morphogenesis Mus Mutation Pathogenesis Patients Perinatal mortality demographics Point Mutation Proteins Pulmonary veins Regulation Reporting Respiratory Insufficiency Role STAT3 gene Severities Signal Transduction Testing Therapeutic Transgenes Transgenic Mice Transgenic Organisms Vascular Endothelial Growth Factors angiogenesis base capillary clinically relevant effective therapy efficacy testing gain of function improved in vivo insight knock-down loss of function migration mortality mutant neonate novel novel therapeutic intervention prevent promoter protein function protein protein interaction research study small molecule transcription factor

项目摘要

DESCRIPTION (provided by applicant): Alveolar Capillary Dysplasia with Misalignment of Pulmonary Veins (ACD/MPV) is a lethal congenital disorder of neonates and infants caused by severe defects in development of pulmonary vasculature. Due to the severity of developmental defects and progressive respiratory insufficiency in ACD/MPV infants, mortality usually occurs within the first month of birth. There is no effective treatment for ACD/MPV patients, and new therapeutic approaches are greatly needed. Recently, dominantly inherited heterozygous mutations in the Forkhead Box F1 (FOXF1) gene were found in 40% of ACD/MPV cases. My laboratory previously demonstrated that FOXF1 transcription factor is a critical regulator of pulmonary vascular development. Global deletion of the mouse Foxf1 gene is embryonic lethal, whereas mice heterozygous for the Foxf1-null allele (Foxf1+/-) had reduced numbers of pulmonary capillaries, lung hypoplasia and increased perinatal mortality, all key features of human ACD/MPV. During the previous funding period, we generated mice with endothelial-specific deletion of Foxf1 and demonstrated that FOXF1 acts in cell autonomous manner to regulate angiogenesis and VEGF signaling in pulmonary endothelial cells. While 42 distinct FOXF1 mutations have been linked to ACD/MPV, molecular mechanisms by which these mutations perturb pulmonary morphogenesis and function remain unknown. We will focus on five distinct FOXF1 mutations that may have different pathogenesis in ACD/MPV. Our hypothesis is that FOXF1 mutations disrupt lung vascular development by inhibiting DNA binding (S52F and G91E mutations), interfering with function of wild type (WT) FOXF1 protein (Del872-879) or inactivating STAT3 signaling (Y284A and I285Q). In Aim 1, we will introduce the FOXF1 mutations into primary lung endothelial cells to determine molecular mechanisms by which these mutations disrupt cellular proliferation, migration and angiogenesis. We will also generate knock-in mice expressing one WT Foxf1 allele and one mutant Foxf1 allele, mimicking genetic abnormalities in ACD/MPV patients. We will use these mice to identify molecular mechanisms by which the FOXF1 mutations perturb pulmonary morphogenesis and function. We will also use a novel small molecule compound, which stabilizes WT FOXF1 protein, to stimulate development of pulmonary vasculature and improve survival in mouse ACD models. In Aim 2, using mass spectrometry we have already found that FOXF1 directly binds to the STAT3 protein, a key transcriptional regulator of endothelial cells. Thus, we will determine if FOXF1 functions as a co- activator of STAT3 to induce STAT3 signaling. We will also test whether disruption of FOXF1-STAT3 interactions by Y284A and I285Q FOXF1 mutations impairs pulmonary vascular development in vivo. Altogether, since reduced FOXF1 and STAT3 have been reported in several lung diseases, FOXF1-stabilizing compounds, which were recently identified in my laboratory, could have broad therapeutic applications for treatment of currently incurable ACD/MPV and other lung diseases associated with vascular insufficiency.

描述（由申请人提供）：肺泡毛细血管发育不良伴肺静脉错位（ACD/MPV）是一种致命的新生儿和婴儿先天性疾病，由严重的肺血管发育缺陷和进行性呼吸功能不全引起。 ACD/MPV 婴儿的死亡通常发生在出生第一个月内。 ACD/MPV 患者尚无有效的治疗方法，需要新的治疗方法。最近，我的实验室在 40% 的 ACD/MPV 病例中发现了 Forkhead Box F1 (FOXF1) 基因的显性遗传杂合突变，FOXF1 转录因子是肺血管发育的关键调节因子。小鼠 Foxf1 基因具有胚胎致死性，而 Foxf1 缺失等位基因 (Foxf1+/-) 杂合的小鼠肺毛细血管数量减少、肺发育不全，肺毛细血管数量增加。围产期死亡率，人类 ACD/MPV 的所有关键特征在之前的资助期间，我们培育了内皮特异性缺失 Foxf1 的小鼠，并证明 FOXF1 以细胞自主方式调节肺内皮细胞中的血管生成和 VEGF 信号传导。不同的 FOXF1 突变已与 ACD/MPV 相关，但这些突变扰乱肺形态发生和功能的分子机制仍然未知，我们将重点关注可能与 ACD/MPV 相关的五种不同的 FOXF1 突变。我们的假设是，FOXF1 突变通过抑制 DNA 结合（S52F 和 G91E 突变）、干扰野生型 (WT) FOXF1 蛋白 (Del872-879) 的功能或失活 STAT3 信号传导 (Y284A) 来破坏肺血管发育。在目标 1 中，我们将 FOXF1 突变引入原代肺内皮细胞以确定分子机制。我们还将产生表达一种 WT Foxf1 等位基因和一种突变型 Foxf1 等位基因的敲入小鼠，模拟 ACD/MPV 患者的遗传异常。我们还将使用一种新型小分子化合物来稳定 WT FOXF1 蛋白，以刺激肺血管系统的发育并提高小鼠 ACD 的存活率。在目标 2 中，我们使用质谱法发现 FOXF1 直接与内皮细胞的关键转录调节因子 STAT3 蛋白结合，因此，我们将确定 FOXF1 是否作为 STAT3 的共激活剂来诱导 STAT3 信号传导。我们还将测试 Y284A 和 I285Q FOXF1 突变对 FOXF1-STAT3 相互作用的破坏是否会损害体内肺血管发育。由于已报道几种肺部疾病中 FOXF1 和 STAT3 减少，最近在我的实验室中鉴定出的 FOXF1 稳定化合物可能具有广泛的治疗应用，可用于治疗目前无法治愈的 ACD/MPV 和其他与血管功能不全相关的肺部疾病。