Novel Molecular Mechanisms Regulating Postnatal Pulmonary Angiogenesis

调节产后肺血管生成的新分子机制

• 批准号: 9265926
• 负责人: Cristina Maria Alvira
• 金额: $ 48.26万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-02 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9265926
• 关键词: Address; Adult; Affect; Air Sacs; Alveolar; Angiogenic Factor; Animal Model; Area; Blood Vessels; Blood capillaries; Bronchopulmonary Dysplasia; Cell Maturation; Cell Survival; Complication; Data; Development; Disease; Distal; Endothelial Cells; Endothelium; Gases; Gene Expression Profile; Growth; Impairment; In Vitro; Infant; Inflammation; Injury; Knowledge; Link; Lung; Lung diseases; Malignant Neoplasms; Mediating; Mediator of activation protein; Molecular; Mus; NF-kappa B; Natural regeneration; Neonatal; Organ Culture Techniques; Pathogenesis; Pathway interactions; Patients; Phenotype; Physiological; Premature Birth; Process; Proteins; Proteomics; Role; Surface; Testing; Therapeutic; Transforming Growth Factors; Translating; Tumor Angiogenesis; Vascularization; Work; Wound Healing; alpha-Fetoproteins; angiogenesis; blood vessel development; capillary; cell motility; density; experimental study; in vivo; innovation; loss of function; lung development; migration; novel; novel therapeutic intervention; postnatal; public health relevance; response to injury; targeted treatment; transcription factor

DESCRIPTION (provided by applicant): Growth of the pulmonary capillaries by angiogenesis is essential for alveolarization, and disruption of pulmonary angiogenesis contributes to the pathogenesis of bronchopulmonary dysplasia (BPD), the most common complication of premature birth. However, the molecular pathways that orchestrate pulmonary angiogenesis during development and disease are not fully understood. This gap in knowledge continues to confound efforts to develop targeted therapies to treat lung diseases caused by impaired angiogenesis, including BPD. Nuclear factor κ-B (NFκB) is a key regulator of inflammation, and growing evidence suggests that NFκB regulates angiogenesis in wound healing in cancer. In contrast, little is known about the role of NFκB in the developing lung. We recently demonstrated that NFκB is essential for alveolarization. NFκB is constitutively active in the neonatal, but quiescent in the adult lung, and inhibiting NFκB disrupts pulmonary angiogenesis and alveolarization in neonatal mice, but has no effect in adults. Blocking NFκB in neonatal primary pulmonary endothelial cells (PEC) impairs survival, proliferation, and in vitro angiogenesis. Activation of NFκB in the PEC corresponds to the period of angiogenesis in the developing lung, peaking at the onset of alveolarization, and decreasing to low levels by mid-alveolarization. However, the capacity for NFκB to be activated in the PEC is not intrinsic to the cell's maturation, as conditioned media obtained from early alveolar lung organ culture (EA-LCM), robustly activates NFκB and enhances migration in adult PEC. Global proteomic analysis to compare the early alveolar and adult lung secretomes identified factors secreted only by the early alveolar lung (EAL). Studies to examine the effect of neutralizing each of these proteins on the ability of the EA-LCM to promote NFκB activation and migration, allowed the identification of two putative activators of NFκB in the early alveolar lung: transforming growth factor-ß induced protein (TGFBI) and alpha-fetoprotein (AFP). Both factors are expressed in the EAL in vivo but minimally expressed in the adult lung, and both are suppressed by injuries that disrupt alveolarization. Therefore, our data suggest the overall working hypothesis that factors uniquely secreted by the EAL promote lung angiogenesis and alveolarization by activating NFκB in the pulmonary endothelium by addressing two inter-related specific aims. In Aim 1, we will examine if TGFBI and AFP secretion by the EAL induces a pro-angiogenic phenotype in the PEC via an NFκB-dependent pathway, and explore if TGFBI and AFP activate common or complementary patterns of gene expression. Aim 2 will utilize in vivo gain and loss of function strategies to determine if disrupting TGFBI- or AFP-mediated activation of NFκB impairs alveolar growth during development and injury. The successful completion of these studies will establish a novel, physiologic role for NFκB in the developing lung, and identify new angiogenic factors that can be directly translated into therapeutic strategies to promote lung growth and regeneration in diseases marked by impaired pulmonary angiogenesis.

描述（由申请人提供）：血管生成引起的肺毛细血管生长对于肺泡化至关重要，肺血管生成的破坏导致支气管肺发育不良（BPD）的发病机制，这是早产最常见的并发症。发育和疾病过程中的肺血管生成尚不完全清楚，这种知识上的差距继续困扰着开发靶向疗法来治疗由血管生成受损引起的肺部疾病的努力，包括。 BPD。核因子 κ-B (NFκB) 是炎症的关键调节因子，越来越多的证据表明 NFκB 在癌症伤口愈合中调节血管生成，而我们最近证明了 NFκB 在肺部发育中的作用。 NFκB 对于肺泡化至关重要。 NFκB 在新生小鼠中持续活跃，但在成年小鼠肺中处于静止状态，抑制 NFκB 会破坏新生小鼠的肺血管生成和肺泡化，但对成年小鼠的原代肺内皮细胞 (PEC) 阻断 NFκB 则没有影响。 PEC 中 NFκB 的激活对应于肺发育中的血管生成时期，峰值为然而，NFκB 在 PEC 中被激活的能力并不是细胞成熟所固有的，因为条件培养基来自早期肺泡肺器官培养物 (EA-LCM)。 ，强力激活 NFκB 并增强成人 PEC 中的迁移，以比较早期肺泡和成人肺分泌组，确定仅由早期肺泡肺分泌的因子。 (EAL). 研究中和这些蛋白质对 EA-LCM 促进 NFκB 激活和迁移的能力的影响，确定了早期肺泡肺中两种假定的 NFκB 激活剂：转化生长因子-ß。诱导蛋白（TGFBI）和甲胎蛋白（AFP）在体内 EAL 中表达，但在成人肺中表达最少，并且都受到破坏肺泡化的损伤的抑制。数据表明总体工作假设是，EAL 独特分泌的因子通过激活肺内皮中的 NFκB 来促进肺血管生成和肺泡化，通过解决两个相互关联的特定目标，我们将检查 EAL 是否诱导 TGFBI 和 AFP 分泌。通过 NFκB 依赖性途径在 PEC 中形成促血管生成表型，并探索 TGFBI 和 AFP 是否会激活基因表达的共同或互补模式 Aim 2。体内功能获得和丧失策略，以确定破坏 TGFBI 或 AFP 介导的 NFκB 激活是否会损害发育和损伤过程中的肺泡生长。这些研究的成功完成将确定 NFκB 在发育中的肺中的新生理作用。确定新的血管生成因子，这些因子可以直接转化为治疗策略，以促进以肺血管生成受损为特征的疾病的肺生长和再生。