Development of novel therapeutic approaches for treatment of Alveolar Capillary Dysplasia

开发治疗肺泡毛细血管发育不良的新疗法

基本信息

批准号：
10218489
负责人：
Tanya Kalin
金额：
$ 53.95万
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：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10218489
关键词：
ACVRL1 gene Alleles Alveolar Alveolar capillary dysplasia with misalignment of pulmonary veins Applications Grants Birth Blood Circulation Blood capillaries Bronchopulmonary Dysplasia Cell Lineage Cell Proliferation Cell Therapy Cell Transplantation Cells Complementary DNA Congenital Disorders Congenital alveolar dysplasia Cyanosis Data Development Diaphragmatic Hernia Diffuse Disease Donor person Embryo Endothelial Cells Endothelium Engraftment Exhibits FOXF1 gene Gene Delivery Genes Genetic Transcription Hemorrhage Heterogeneity Human In Vitro Infant Inherited Laboratories Life Link LoxP-flanked allele Lung Lung Transplantation Modeling Mus Mutation Neonatal Newborn Infant PECAM1 gene PTPRC gene Pathologic Pathway interactions Patients Prognosis Protocols documentation Publishing Pulmonary Circulation Pulmonary veins Resistance Respiratory Failure Respiratory Insufficiency Respiratory physiology STAT3 gene Signal Pathway Signal Transduction Stem cell transplant TEK gene Testing Therapeutic Transgenic Mice Transplantation Vascular Endothelial Growth Factors angiogenesis base bone morphogenetic protein 9 c-myc Genes clinically relevant density effective therapy embryonic stem cell endothelial stem cell expression vector gene delivery system genomic locus human embryonic stem cell improved in vivo innovation lung development mortality mouse model nanoparticle nanoparticle delivery neonate novel novel therapeutic intervention postnatal preservation prevent pulmonary arterial hypertension respiratory self-renewal single-cell RNA sequencing transcription factor

项目摘要

PROJECT SUMMARY. Alveolar Capillary Dysplasia with Misalignment of Pulmonary Veins (ACDMPV) is a fatal congenital disorder of neonates and infants which leads to respiratory insufficiency due to reduced numbers of alveolar capillaries and is associated with lung hypoplasia, hemorrhage, malposition of pulmonary veins and pulmonary arterial hypertension (PAH). ACDMPV is linked to mutations in the Forkhead Box F1 (FOXF1) gene and is resistant to all available therapies and respiratory support, causing cyanosis and respiratory failure in the first month after birth. In rare cases, ACDMPV patients survive several months or even years, but they require lung transplantation early in life. Given the lack of effective treatments for ACDMPV, there is an urgent need for innovative therapeutic approaches to stimulate pulmonary angiogenesis and preserve respiratory function in ACDMPV infants. In our preliminary data, we have generated a clinically relevant model of ACDMPV by introducing the S52F FOXF1 mutation (found in ACDMPV patients) into the endogenous mouse Foxf1 gene locus. Foxf1WT/S52F newborn mice exhibited alveolar capillary dysplasia, misalignment of pulmonary veins, PAH, and increased mortality, all key features of human ACDMPV. Endothelial proliferation and STAT3 signaling were decreased in Foxf1WT/S52F mice and human ACDMPV lungs. In the present grant application, we will test the hypothesis that increasing neonatal lung angiogenesis will decrease PAH, improve survival and prevent lung remodeling in mouse ACDMPV models. In Aim 1, we have developed a novel nanoparticle gene delivery system targeting >85% of pulmonary endothelial cells in vivo when delivered into the neonatal blood circulation. Nanoparticle delivery of STAT3 cDNA after birth induced endothelial proliferation and increased alveolar microvascular density in Foxf1WT/S52F neonatal lungs. We will use two mouse models of ACDMPV (Foxf1WT/S52F and Foxf1+/-) to test whether nanoparticle delivery of STAT3 or FOXF1 will decrease PAH, improve survival and prevent lung remodeling. We will also determine whether the FOXF1/STAT3/cMYC transcriptional cascade is required for neonatal lung angiogenesis. In Aim 2, we provided preliminary data demonstrating that a cell transplantation with pulmonary endothelial progenitor cells (EPCs) (FOXF1+cKit+CD31+CD45-) increases the capillary density in Foxf1WT/S52F lungs. We will investigate heterogeneity of pulmonary EPCs and test their therapeutic potential in mouse ACDMPV models. We will test if EPCs stimulate neonatal lung angiogenesis via the BMP-9/ACVRL1 signaling pathway. Finally, we will use a novel protocol for in vitro differentiation of EPCs from mouse and human embryonic stem cells (ES) and determine if a cell therapy with ES-derived EPCs will be beneficial in mouse ACDMPV models. Altogether, the proposed studies will directly test whether endothelial delivery of STAT3 or cell therapy with EPCs have therapeutic potential in ACDMPV.

项目摘要。肺泡毛细血管发育不良伴肺静脉错位 (ACDMPV) 是一种新生儿和婴儿致命的先天性疾病，由于呼吸功能减少而导致呼吸功能不全肺泡毛细血管的数量，与肺发育不全、出血、肺错位有关静脉和肺动脉高压（PAH）。 ACDMPV 与 Forkhead Box F1 突变有关 (FOXF1) 基因，对所有可用的治疗和呼吸支持都有抵抗力，导致发绀和出生后第一个月呼吸衰竭。在极少数情况下，ACDMPV 患者可以存活数月甚至数月岁，但他们在生命早期就需要进行肺移植。鉴于 ACDMPV 缺乏有效的治疗方法，迫切需要创新的治疗方法来刺激肺血管生成和保护 ACDMPV 婴儿的呼吸功能。在我们的初步数据中，我们已经生成了临床通过将 S52F FOXF1 突变（在 ACDMPV 患者中发现）引入到 ACDMPV 的相关模型中内源性小鼠 Foxf1 基因位点。 Foxf1WT/S52F新生小鼠表现出肺泡毛细血管发育不良，肺静脉错位、PAH 和死亡率增加，这些都是人类 ACDMPV 的关键特征。 Foxf1WT/S52F 小鼠和人 ACDMPV 的内皮增殖和 STAT3 信号传导减少肺。在本拨款申请中，我们将检验以下假设：增加新生儿肺血管生成将减少小鼠 ACDMPV 模型中的 PAH、提高存活率并预防肺重塑。在目标 1 中，我们开发了一种新型纳米颗粒基因传递系统，靶向 85% 以上的肺内皮细胞当输送到新生儿血液循环中时体内。出生后 STAT3 cDNA 的纳米颗粒递送 Foxf1WT/S52F 新生儿肺中诱导内皮增殖并增加肺泡微血管密度。我们将使用两种 ACDMPV 小鼠模型（Foxf1WT/S52F 和 Foxf1+/-）来测试纳米粒子是否递送 STAT3 或 FOXF1 将减少 PAH、提高生存率并防止肺重塑。我们还将确定 FOXF1/STAT3/cMYC 转录级联是否是新生儿肺血管生成所必需的。瞄准 2、我们提供的初步数据表明肺内皮祖细胞移植细胞 (EPC) (FOXF1+cKit+CD31+CD45-) 增加 Foxf1WT/S52F 肺中的毛细血管密度。我们将研究肺 EPC 的异质性并测试其在小鼠 ACDMPV 模型中的治疗潜力。我们将测试 EPC 是否通过 BMP-9/ACVRL1 信号通路刺激新生儿肺血管生成。最后，我们将使用一种新的方案对小鼠和人类胚胎干细胞中的 EPC 进行体外分化 (ES) 并确定使用 ES 衍生的 EPC 进行细胞疗法是否对小鼠 ACDMPV 模型有益。总而言之，拟议的研究将直接测试 STAT3 的内皮传递或细胞疗法是否有效 EPCs 在 ACDMPV 中具有治疗潜力。