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（由申请人提供）：肺毛细血管的血管生成对肺泡形成至关重要，而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，这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不完全清楚。这种知识上的差距继续困扰着开发靶向治疗方法来治疗由血管生成受损引起的肺部疾病的努力，包括BPD。核因素？(NF - b吗?B)是炎症的关键调节因子，越来越多的证据表明NF？B调节肿瘤创面愈合过程中的血管生成。相比之下，人们对NF的作用知之甚少。B在发育中的肺中。我们最近证明了NF？B是肺泡形成所必需的。NF吗?B在新生儿中具有组成性活性，但在成人肺中处于静止状态，并抑制NF？B破坏新生小鼠的肺血管生成和肺泡形成，但对成年小鼠没有影响。阻塞NF吗?新生儿原发性肺内皮细胞（PEC）中的B损害存活、增殖和体外血管生成。活化NF？PEC中的B与发育中的肺血管生成时期相对应，在肺泡化开始时达到峰值，在肺泡化中期下降到较低水平。然而，NF的容量？B在PEC中被激活并不是细胞成熟所固有的，因为从早期肺泡肺器官培养（EA-LCM）中获得的条件培养基强有力地激活了NF？B并增强成人PEC的迁移。通过全球蛋白质组学分析比较早期肺泡肺和成人肺分泌组，确定了仅由早期肺泡肺（EAL）分泌的因子。研究这些蛋白的中和作用对EA-LCM促进NF的能力的影响？B激活和迁移，允许鉴定两种假定的NF？早期肺泡肺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中均有表达，但在成人肺中表达极少，且均被破坏肺泡化的损伤所抑制。因此，我们的数据表明，EAL分泌的独特因子通过激活NF - 3来促进肺血管生成和肺泡形成。B通过解决两个相互关联的特定目的在肺内皮。在Aim 1中，我们将检验EAL分泌的TGFBI和AFP是否通过NF诱导PEC中促血管生成表型？并探讨TGFBI和AFP是否激活了共同或互补的基因表达模式。目的2将利用体内功能的获得和丧失策略来确定是否破坏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确立NF的一种新的生理作用。并发现新的血管生成因子，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标志的疾病的肺生长和再生。