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

肺泡上皮细胞源性细胞通讯网络因子 2 (CCN2) 在肺泡发生和支气管肺发育不良中的作用

• 批准号: 10684698
• 负责人: Deepthi Alapati
• 金额: $ 16.2万
• 依托单位: ALFRED I. DU PONT HOSP FOR CHILDREN
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10684698
• 关键词: 3-Dimensional; Abnormal Cell; Address; Adverse effects; Affect; Alveolar; Animal Model; Attenuated; Basic Science; Bioinformatics; Biological Assay; Bronchopulmonary Dysplasia; CRISPR/Cas technology; Candidate Disease Gene; Cause of Death; Cell Communication; Cell Lineage; Cell physiology; Cells; Characteristics; Chronic lung disease; Clinical; Coculture Techniques; Communication; Complex; DNA; Data Set; Disease; Endothelial Cells; Endothelium; Environment; Epithelial Cells; Epithelium; Equilibrium; Exposure to; Extracellular Matrix; Fibroblasts; Future; Gene Expression; Genes; Genetic; Goals; Hyperoxia; Impairment; In Vitro; Infant; Integrins; International; Knock-out; Knockout Mice; Label; Laboratories; Lung; Lung diseases; Master of Science; Mediating; Mentors; Mentorship; Mesenchymal; Modeling; Modification; Molecular; Monoclonal Antibodies; Morbidity - disease rate; Mutation; Neonatal; Neurodevelopmental Impairment; Outcome; Pathologic Processes; Pathway interactions; Pediatric Hospitals; Pennsylvania; Phenotype; Premature Infant; Prevention; Prevention therapy; Production; Productivity; Proteins; Rattus; Reagent; Repression; Research; Research Personnel; Research Proposals; Resources; Role; Route; Scientist; Signal Transduction; System; Technology; Therapeutic; Therapeutic Agents; Time; Training; Training Programs; Transgenic Organisms; Translational Research; Universities; Vascularization; Wild Type Mouse; alveolar epithelium; antagonist; career; cell type; clinical care; collaborative environment; combinatorial; effective therapy; experience; experimental study; genetic manipulation; hospital readmission; hyperoxia induced lung injury; improved; in utero; innovation; insight; knock-down; lung development; lung injury; lung regeneration; lung repair; mortality; mouse model; new therapeutic target; novel; novel strategies; novel therapeutics; overexpression; postnatal; prevent; response; selective expression; single-cell RNA sequencing; skills; spatiotemporal; targeted treatment; therapeutic genome editing; tool

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.

项目概要/摘要 该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、理学硕士、主治医师）提供培训 Nemours AI 杜邦儿童医院的新生儿科医生提供指导、培训和研究 成为独立临床科学家和新生儿肺病领导者所需的经验 研究。她获得了宾夕法尼亚大学转化研究硕士学位。 候选人的长期职业目标是开发新疗法来促进肺部修复和再生 治疗新生儿肺部疾病，例如支气管肺发育不良（BPD）。为了实现这些目标，她 将由国际公认的肺基础科学研究专家团队指导 内穆尔和宾夕法尼亚大学与她有着良好的指导记录 和生产力。她将接受严格的教学和实践培训计划，以获得以下方面的专业知识 使用复杂的方法阐明肺微环境内肺细胞特异性分子机制 遗传小鼠模型和 3D 共培养系统；基因编辑技术作为机制的应用 调节 BPD 等复杂肺部疾病候选基因的治疗工具；和专业知识 先进的生物信息学。她的研究重点是（1）阐明细胞通信的功能 肺泡发生期间肺泡上皮细胞 (AEC) 中的网络因子 2 (Ccn2) 以及 (2) 评估是否 调节 AEC 中 Ccn2 的表达将改善高氧诱导的 BPD 表型。本研究提案 建立在候选人的初步研究基础上，该研究表明 Ccn2 在严重的疾病中具有重要的致病作用 BPD 和 CRISPR-Cas9 基因编辑技术操纵疾病的治疗潜力 在发育中的肺上皮中表达的基因。 BPD 是最常见的死亡原因，严重者 神经发育障碍和早产儿再入院。尽管临床取得了进步 护理方面，预防和治疗 BPD 的临床努力基本上不成功。作为最常接触的细胞类型 在外部环境中，AEC 已成为许多肺部疾病（包括 BPD）的焦点。 此外，AEC 可以通过羊膜内、鼻内和腔内递送的治疗剂直接靶向。 气管内途径。因此，更好地了解 AEC 期间驱动肺泡生成的机制 正常的肺部发育和对产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。通过精确检查 Ccn2 在 AEC 过程中的空间和时间功能 肺泡发生和高氧引起的早期产后肺损伤，这项研究将提供新的机制 深入了解其在调节肺泡生成中的作用，并开发出针对 AEC 衍生的 Ccn2 的新疗法 严重 BPD 的预防和治疗。候选人可以使用所有必需的技术、实验室和 理想的协作环境中的智力资源，为成功开展研究奠定了基础 独立调查员的发展之路。

项目成果

Pulmonary immune cell transcriptome changes in double-hit model of BPD induced by chorioamnionitis and postnatal hyperoxia.

• DOI: 10.1038/s41390-020-01319-z
• 发表时间: 2021-09
• 期刊: Pediatric research
• 影响因子: 3.6
• 作者: Shrestha D;Ye GX;Stabley D;Betal SGN;Zhu Y;Glazewski L;Holbrook J;Sethi M;Hesek A;Shaffer TH;Aghai ZH;Addya S;Alapati D
• 通讯作者: Alapati D

Administration of Drugs/Gene Products to the Respiratory System: A Historical Perspective of the Use of Inert Liquids.

• DOI: 10.3389/fphys.2022.871893
• 发表时间: 2022
• 期刊: FRONTIERS IN PHYSIOLOGY
• 影响因子: 4
• 作者: Alapati, Deepthi;Shaffer, Thomas H.
• 通讯作者: Shaffer, Thomas H.