Rebuilding the Lung

描述(由申请人提供)：我们的最终目标是使用去细胞肺作为肺前体细胞的自然三维(3-D)基质基础来重建肺。肺移植通常是不可逆的结构性肺损伤患者的唯一选择，但受到主要障碍的阻碍，如缺乏肺器官捐赠者和慢性排斥反应。人工支架不能复制肺的分支或细胞外基质的组成、排列和拉伸。我们将专注于利用通风的、去细胞的全肺作为自然的3-D基质来重建肺，并用具有肺上皮祖细胞和内皮祖细胞潜能的细胞重新填充它。我们有数据表明，啮齿动物的肺可以成功地去细胞，同时保持细胞外基质在正确的地理空间分支模式，即使在长时间的呼吸。成人和胎儿的肺细胞可以成功地在脱细胞的、通风的肺中输注和培养。我们假设，在适当的生长条件下，假定的祖细胞将重新上皮化和重新内皮化基质，并产生肺细胞特有的蛋白质。我们还预计，细胞输入的时机、生长因子和发育阶段适当的呼吸拉伸将对这一过程至关重要。在目标1中，我们将确定可能的肺祖细胞群，如诱导多能(IPS)细胞、骨髓来源细胞和EpCAM+肺来源细胞，是否会在我们新的“肺生物反应器”系统中的通风、脱细胞、全肺基质的环境中生长和分化。我们将比较在成人肺和胎儿肺制备的脱细胞基质中单独和联合培养这些细胞。在目标2中，我们将确定是否可以使用血液生长内皮细胞(BOECs)和目标1中最好的细胞来重新分化上皮和内皮。我们处于进行这些研究的独特位置。我们有一个独特的模式系统和经验丰富的合作者，他们为这项赠款申请提供新颖的细胞、关键的专业知识和建议。 公共卫生相关性：肺移植通常是患有不可逆转的结构性肺损伤的患者的唯一选择，但受到主要障碍的阻碍，如缺乏肺器官捐赠者和慢性排斥反应。我们将专注于使用通风的、脱细胞的整个啮齿动物肺作为自然的3-D基质来重建肺，并用具有向肺组织生长的潜力的细胞重新填充它，并产生肺细胞特有的蛋白质。

DESCRIPTION (provided by applicant): Our ultimate goal is to rebuild the lung using decellularized lung as a natural 3-dimensional (3-D) matrix base for lung progenitor cells. Lung transplantation is usually the only option for patients with irreversible structural lung damage but is hindered by major obstacles such as lack of lung organ donors and chronic rejection. Artificial scaffolds cannot replicate the branching or the composition, arrangement and stretch of the extracellular matrix of the lung. We will focus on rebuilding the lung using ventilated, decellularized whole lungs as a natural 3-D matrix and repopulate it with cells having potential as lung epithelial and endothelial progenitors. We have data that rodent lungs can successfully be decellularized while maintaining extracellular matrix in the correct geospatial branching pattern even after prolonged ventilation. Adult and fetal lung cells can successfully be infused and cultured in decellularized, ventilated lungs. We hypothesize that, given the right growth conditions, putative progenitors will re-epithelialize and re-endothelialize the matrix and produce proteins characteristic of lung cells. We also expect that timing of cell input, growth factors and developmental stage-appropriate ventilatory stretch will be critical to this process. In Aim 1, we will determine if putative lung progenitor cell populations such as induced pluripotent (iPS) cells, BM-derived cells and EpCAM+ lung-derived cells, will grow and differentiate in the setting of a ventilated, decellularized, whole lung matrix in our new "lung bioreactor" system. We will compare culturing these cells, alone and in combination, in decellularized matrices prepared from adult and fetal lungs. In Aim 2, we will determine if both epithelium and endothelium can be recellularized using blood outgrowth endothelial cells (BOECs) and the best cells from Aim 1. We are in a unique position to perform these studies. We have a unique model system and highly experienced collaborators who provide novel cells, critical expertise and advice for this grant application. PUBLIC HEALTH RELEVANCE: Lung transplantation is usually the only option for patients with irreversible structural lung damage but is hindered by major obstacles such as lack of lung organ donors and chronic rejection. We will focus on rebuilding the lung using ventilated, decellularized whole rodent lungs as a natural 3-D matrix and repopulate it with cells having the potential to grow into lung tissue and produce proteins characteristic of lung cells.