Role of alveolar epithelial cell-derived cellular communication network factor 2 (CCN2) in alveologenesis and bronchopulmonary dysplasia

项目摘要/摘要 本提案描述了一项为期5年的培训计划，旨在为Deepthi Alapati博士、医学博士、医学硕士提供 Nemour AI DuPont儿童医院的新生儿专家与导师、培训和研究 成为一名独立的临床科学家和新生儿肺部疾病领导者所需的经验 研究。她获得了宾夕法尼亚大学翻译研究硕士学位。 候选人的长期职业目标是开发新的疗法，以促进肺修复和再生 治疗新生儿肺部疾病，如支气管肺发育不良(BPD)。为了实现这些目标，她 将由一个国际公认的肺基础科学研究专家团队指导 Nemour和宾夕法尼亚大学，她在这两所学校有成功的指导记录 和生产力。她将接受严格的教学和实践培训计划，以获得专业知识 应用复合体阐明肺微环境中肺细胞特异性分子机制 遗传小鼠模型和3D共培养系统；作为机制的基因编辑技术的应用 和治疗工具，用于调节BPD等复杂肺部疾病的候选基因；以及 先进生物信息学。她的研究将集中在(1)阐明细胞通讯的功能 肺泡上皮细胞(AEC)中网络因子2(Ccn2)在肺泡形成过程中的表达 调节AEC中Ccn2的表达将改善高氧诱导的BPD表型。这项研究建议 是建立在候选人的初步研究基础上的，该研究表明Ccn2在严重的 BPD和CRISPR-Cas9基因编辑技术操纵致病的治疗潜力 基因在发育中的肺上皮细胞中表达。BPD是最常见的死亡原因，严重 早产儿的神经发育障碍和再次住院。尽管在临床上取得了进步 护理、临床预防和治疗BPD的努力大多不成功。作为接触最多的细胞类型 在外部环境方面，AEC已成为包括BPD在内的许多肺部疾病的中心焦点。 此外，AEC可以直接作为治疗药物的靶点，通过羊水、鼻腔和 气管内路径。因此，更好地了解在AEC中驱动肺泡形成的机制 正常的肺发育和对出生后早期肺损伤的反应将为靶向治疗铺平道路 对于严重的BPD。通过精确检测Ccn2在AEC中的时空功能 高氧诱导的肺泡生成和早期肺损伤，本研究将提供新的机制 洞察其在调节肺泡生成中的作用并导致针对AEC来源的Ccn2的新疗法 重症BPD的防治。应聘者有权访问所有必需的技术、实验室和 在理想的协作环境中提供智力资源，以成功开展研究和铺平道路 作为一名独立调查员的发展之路。

PROJECT SUMMARY/ABSTRACT This proposal describes a 5-year training program to provide Dr. Deepthi Alapati, MD, MS, Attending Neonatologist at Nemours AI duPont Hospital for Children with the mentorship, training and research experience required to become an independent clinician scientist and a leader in neonatal lung disease research. She has received a Master’s degree in Translational Research from University of Pennsylvania. Candidate’s long-term career goal is to develop novel therapies to promote lung repair and regeneration for treatment of neonatal lung diseases such as bronchopulmonary dysplasia (BPD). To achieve these goals, she will be mentored by a team of internationally recognized experts in pulmonary basic science research from Nemours and University of Pennsylvania with whom she has a proven track record of successful mentorship and productivity. She will undergo a rigorous didactics and hands-on training program to acquire expertise in elucidating pulmonary cell-specific molecular mechanisms within the lung microenvironment using complex genetic mouse models and 3D co-culture systems; application of gene editing technologies as a mechanistic and therapeutic tool to modulate candidate genes in complex lung diseases such as BPD; and expertise in advanced bioinformatics. Her research will focus on (1) elucidating the function of Cellular communication network factor 2 (Ccn2) in alveolar epithelial cells (AEC) during alveologenesis and (2) evaluating whether modulating Ccn2 expression in AEC will improve a hyperoxia-induced BPD phenotype. This research proposal is built upon candidate’s preliminary research that demonstrates an important causative role for Ccn2 in severe BPD and the therapeutic potential of CRISPR-Cas9 gene editing technology to manipulate disease causing genes expressed in the developing pulmonary epithelium. BPD is the most common cause of death, severe neurodevelopmental impairment, and hospital readmissions in preterm infants. In spite of advances in clinical care, clinical efforts to prevent and treat BPD have been largely unsuccessful. As the cell type most exposed to the external environment, AEC have emerged as a central focus in many lung diseases, including BPD. Moreover, AEC can be directly targeted by therapeutic agents delivered through intra-amniotic, intra-nasal and intra-tracheal routes. Thus, a better understanding of mechanisms in AEC that drive alveologenesis during normal lung development and in response to early postnatal lung injury, would pave path for targeted therapies for severe BPD. By precisely examining the spatial and temporal function of Ccn2 in AEC during alveologenesis and early postnatal lung injury induced by hyperoxia, this study will provide novel mechanistic insights into its role in regulating alveologenesis and lead to novel therapies targeted at AEC-derived Ccn2 for the prevention and treatment of severe BPD. Candidate has access to all required technical, laboratory and intellectual resources in an ideal collaborative environment for successfully conducting her research and pave the path to develop as an independent investigator.