Bioengineering a Chimeric Human Lung

描述(申请人提供)：美国有近2500万人患有终末期肺部疾病。肺移植是这些患者的唯一治疗选择，但由于供体器官短缺、长期排斥反应和免疫抑制治疗的需要，肺移植仍然受到阻碍。产生含有患者自身细胞的肺的能力将从根本上改变我们目前治疗终末期肺部疾病的方式。我们建议用人类多能干细胞(HPSC)来源的肺祖细胞种植部分脱细胞的供体肺，以构建嵌合的人肺。我们的目标是(1)获得hPSC对肺谱系和肺组织形成的新见解，(2)利用天然肺基质对移植功能肺进行生物工程。我们设想，通过用患者的hPSC来源的肺细胞部分替代细胞材料，可以改善排斥/边缘质量供体肺的功能。我们的假设是，种植到肺脱细胞区域的hPSC前体细胞将被诱导再生和重塑供体肺，以响应来自组织基质和残留细胞的部位特异性信号。我们的方法是取被拒绝移植的供体肺，从肺的有限区域移除细胞，同时保留脱细胞基质和周围完整实质的成分、结构和机械性能。通过同时通过肺实质灌流去细胞液体和通过门静脉注入Perfadex溶液，我们将保存完整的肺血管。然后，脱细胞区域将被hPSC来源的肺前体细胞重新填充，以产生能够进行最低限度可接受的气体交换的肺，这种气体交换将在重塑后改善。目的1是从hPSCs中分离和鉴定不同的肺和呼吸道上皮细胞系。目的二是将人PSC来源的肺细胞接种于脱细胞基质切片上，构建人肺组织生物工程模型。目的3是通过用hPSC来源的肺细胞重新填充肺的去细胞区域形成嵌合的人肺，并使用临床肺灌流系统研究功能的恢复。这项建议的最终目的是通过结合三个主要的创新部分来实现排斥移植的供肺的功能恢复：(1)从hPSC中获得肺细胞，(2)保留血管和实质结构的供体肺的局部去细胞，以及(3)通过将受者的细胞重新植入肺来恢复功能。

DESCRIPTION (provided by applicant): Nearly 25 million people in the United States suffer from end-stage lung disease. Lung transplantation, the only curative option for these patients, remains hampered by donor organ shortages, long-term rejection, and the need for immunosuppressive therapy. The ability to generate lungs containing a patient's own cells would radically change the way we currently treat end-stage lung disease. We propose to bioengineer chimeric human lungs by seeding partially decellularized donor lungs with human pluripotent stem cell (hPSC) derived pulmonary progenitors. Our goals are to (1) obtain new insights into hPSC specification towards pulmonary lineages and the formation of lung tissue using native lung matrix, and (2) bioengineer functional lungs for transplantation. We envision that the function of rejected/marginal quality donor lungs can be improved by partial replacement of cellular material by a patient's hPSC-derived pulmonary cells. Our hypothesis is that hPSC-progenitors seeded into the decellularized regions of the lung will be induced to regenerate and remodel the donor lung in response to site-specific signals from the tissue matrix and residual cells. Our approach will be to take donor lungs rejected for transplantation and remove cells from limited regions of the lung while preserving the composition, architecture, and mechanical properties of the decellularized matrix and the surrounding intact parenchyma. By simultaneously perfusing decellularization fluids through the lung parenchyma and Perfadex solution through the portal vein, we will preserve intact lung vasculature. The decellularized regions will then be repopulated by hPSC-derived lung progentor cells to produce a lung that is capable of some minimally acceptable level of gas exchange that will improve upon remodeling. Aim 1 is to derive and fully characterize the different lineages of the lung and airway epithelium from hPSCs. Aim 2 is to bioengineer human lung tissue by hPSC-derived pulmonary cells cultured on slices of decellularized matrix. Aim 3 is to form a chimeric human lung by repopulating decellularized regions of the lung with hPSC- derived pulmonary cells and investigate functional recovery using a clinical lung perfusion system. This proposal ultimately aims to achieve functional recovery of donor lungs rejected for transplantation by combining three major innovative components: (1) Derivation of pulmonary cells from the hPSCs, (2) Regional decellularization of the donor lung with the preservation of vasculature and parenchymal architecture, and (3) Functional recovery through repopulation of the lung with the recipient's cells.