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

• 批准号: 10895101
• 负责人: Tanya Kalin
• 金额: $ 53.16万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 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/10895101
• 关键词: ACVRL1 gene; Alleles; Alveolar; Alveolar capillary dysplasia with misalignment of pulmonary veins; Applications Grants; Birth; Blood Circulation; Blood Vessels; 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; Heterozygote; 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; Proliferating; 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; Vascular Endothelial Growth Factors; angiogenesis; bone morphogenetic protein 9; c-myc Genes; clinically relevant; density; directed differentiation; effective therapy; embryonic stem cell; endothelial stem cell; expression vector; fibrotic lung; 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

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。肺静脉不对对准肺静脉（ACDMPV）是一个 新生儿和婴儿的致命先天性疾病，导致由于减少而导致呼吸不足 肺泡毛细血管的数量，与肺部发育不全，出血，肺部错位有关 静脉和肺动脉高压（PAH）。 ACDMPV与Forkhead Box F1中的突变有关 （FOXF1）基因，并且对所有可用的疗法和呼吸支持具有抗性，从而导致氰化物和 出生后第一个月的呼吸衰竭。在极少数情况下，ACDMPV患者存活了几个月甚至 多年，但它们需要生命早期的肺移植。鉴于缺乏对ACDMPV的有效治疗 迫切需要创新的治疗方法来刺激肺血管生成和 保留ACDMPV婴儿的呼吸功能。在我们的初步数据中，我们在临床上生成了 通过将S52F FOXF1突变（在ACDMPV患者中发现）引入ACDMPV的相关模型 内源性小鼠FOXF1基因座。 FOXF1WT/S52F新生小鼠表现出肺泡毛细血管发育不良， 肺静脉，PAH和死亡率增加的未对准，这是人ACDMPV的所有关键特征。 FOXF1WT/S52F小鼠和人ACDMPV的内皮增殖和STAT3信号传导降低 肺。在当前的赠款应用中，我们将检验以下假设：新生儿血管生成增加 将减少PAH，提高生存率并防止小鼠ACDMPV模型中的肺重塑。在AIM 1中，我们 已经开发了一种新型的纳米颗粒基因递送系统，其靶向> 85％的肺内皮细胞 当体内输送到新生儿血液循环中时。出生后STAT3 cDNA的纳米颗粒输送 FOXF1WT/S52F新生儿肺中诱导的内皮增殖和肺泡微血管密度增加。 我们将使用两种ACDMPV的鼠标模型（FOXF1WT/S52F和FOXF1 +/-）来测试纳米颗粒是否输送 STAT3或FOXF1将降低PAH，提高生存率并防止肺重塑。我们还将确定 新生儿血管生成是否需要FOXF1/STAT3/CMYC转录级联反应。目标 2，我们提供了初步数据，表明用肺部内皮祖细胞移植细胞移植 细胞（EPC）（FOXF1+CKIT+CD31+CD45-）增加了FOXF1WT/S52F肺的毛细管密度。我们将 研究肺EPC的异质性，并在小鼠ACDMPV模型中测试其治疗潜力。 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将EPC与小鼠和人类胚胎干细胞的体外分化 （ES）并确定具有ES衍生EPC的细胞疗法在小鼠ACDMPV模型中是否有益。 总共提出的研究将直接测试STAT3的内皮递送或使用 EPC在ACDMPV中具有治疗潜力。