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: 8686276
• 负责人: Cristina Maria Alvira
• 金额: $ 43.9万
• 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/8686276
• 关键词: Address; Adult; Affect; Air Sacs; Alveolar; Angiogenic Factor; Animal Model; Area; Blood Vessels; Blood capillaries; Bronchopulmonary Dysplasia; Cell Maturation; Cell Survival; Cells; Complement Factor B; Complication; Conditioned Culture Media; Data; Development; Disease; Distal; Endothelial Cells; Endothelium; Gases; Gene Expression Profile; Growth; Immigration; In Vitro; Infant; Inflammation; Injury; Knowledge; Link; Lung; Lung diseases; Malignant Neoplasms; Mediating; Mediator of activation protein; Molecular; Mus; Natural regeneration; Neonatal; Nuclear; Organ Culture Techniques; Pathogenesis; Pathway interactions; Patients; Phenotype; Physiological; Premature Birth; Process; Proteins; Proteomics; Role; Staging; Surface; TGFBI gene; Testing; Therapeutic; Transforming Growth Factor-Beta Induced Protein IGH3; Transforming Growth Factors; Translating; Tumor Angiogenesis; Vascularization; Work; Wound Healing; alpha-Fetoproteins; angiogenesis; capillary; density; in vivo; innovation; loss of function; lung development; migration; novel; novel therapeutics; postnatal; prevent; public health relevance; research study; response to injury; 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 ?-B (NF?B) is a key regulator of inflammation, and growing evidence suggests that NF?B regulates angiogenesis in wound healing in cancer. In contrast, little is known about the role of NF?B in the developing lung. We recently demonstrated that NF?B is essential for alveolarization. NF?B is constitutively active in the neonatal, but quiescent in the adult lung, and inhibiting NF?B disrupts pulmonary angiogenesis and alveolarization in neonatal mice, but has no effect in adults. Blocking NF?B in neonatal primary pulmonary endothelial cells (PEC) impairs survival, proliferation, and in vitro angiogenesis. Activation of NF?B 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 NF?B 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 NF?B 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 NF?B activation and migration, allowed the identification of two putative activators of NF?B in the early alveolar lung: transforming growth factor-¿ 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 NF? B 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 NF?B-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 NF?B impairs alveolar growth during development and injury. The successful completion of these studies will establish a novel, physiologic role for NF?B 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（由申请方提供）：通过血管生成的肺毛细血管生长对于肺泡化至关重要，并且肺血管生成的破坏有助于支气管肺发育不良（BPD）的发病机制，BPD是早产最常见的并发症。然而，在发育和疾病过程中协调肺血管生成的分子途径尚未完全了解。这种知识上的差距继续混淆了开发靶向疗法以治疗由血管生成受损引起的肺部疾病（包括BPD）的努力。核因素？- B（NF？B）是炎症的关键调节因子，越来越多的证据表明NF？B调节癌症伤口愈合中的血管生成。相比之下，很少有人知道NF的作用？B在发育中的肺。我们最近证明了NF？B对肺泡形成至关重要。NF？B在新生儿肺中是组成性活性的，但在成人肺中是静止的，并且抑制NF？B破坏新生小鼠的肺血管生成和肺泡形成，但对成年小鼠无影响。阻止NF？新生儿原代肺内皮细胞（PEC）中的B损害存活、增殖和体外血管生成。激活NF？PEC中的B对应于发育中的肺中的血管生成期，在肺泡化开始时达到峰值，并在肺泡化中期降至低水平。然而，NF的能力？B被激活的PEC是不是固有的细胞的成熟，作为条件培养基获得早期肺泡肺器官培养（EA-LCM），强烈激活NF？B和增强迁移成人PEC。全球蛋白质组学分析比较早期肺泡和成人肺分泌物组确定的因素分泌的早期肺泡肺（EAL）。研究，以检查中和这些蛋白质的EA-LCM的能力，以促进NF？B激活和迁移，允许确定两个假定的NF激活剂？早期肺泡肺中的B：转化生长因子诱导蛋白（TGFBI）和甲胎蛋白（AFP）。这两种因子在体内EAL中表达，但在成人肺中表达最低，并且两者都受到破坏肺泡化的损伤的抑制。因此，我们的数据表明，整体的工作假设，独特的EAL分泌的因素，促进肺血管生成和肺泡活化NF？B在肺内皮细胞通过解决两个相互关联的具体目标。在目的1中，我们将研究是否TGFBI和AFP分泌的EAL诱导前血管生成表型的PEC通过NF？B依赖的途径，并探讨TGFBI和AFP是否激活共同或互补的基因表达模式。目的2将利用在体内获得和功能的策略，以确定是否破坏TGFBI或AFP介导的NF激活？B在发育和损伤期间损害肺泡生长。这些研究的成功完成将为NF？B在发育中的肺，并确定新的血管生成因子，可以直接转化为治疗策略，以促进肺生长和再生的疾病标志着受损的肺血管生成。