Bioengineering New Lungs from Cadaveric Lung Scaffolds

描述(由申请人提供)：永远不会有足够的供体肺来满足当前和未来的移植需求。相比之下，整个身体肺的去细胞化将产生一个完整的支架，可以用胚胎干细胞或成人干细胞重新细胞化，包括来自个别患者的诱导多能干细胞(IPS)，然后用于自体移植。值得注意的是，去细胞过程去除了导致免疫排斥反应的细胞抗原，去细胞肺保持了天然的呼吸道和肺泡结构、细胞外基质蛋白组成和肺血管网络。这将潜在地无限供应无关的捐赠者身体肺，用于肺气肿和其他目前尚无有效治疗方法的疾病。我们的初步数据表明，去细胞的小鼠肺可以用成体干细胞和胚胎干细胞重新细胞化。此外，当肺重新细胞化时，可以通过组织学和功能结果来评估去细胞肺制剂，包括肺力学、表面活性物质的产生、血管灌流和上皮屏障功能。最重要的是，初步数据表明，在动物模型中，肺可以通过手术重新植入。这些数据为拟议的多机构合作研究提供了坚实的平台，在这些研究中，将利用最先进的生物工程技术来开发在脱细胞肺(包括身体肺)中生长功能性肺组织的最佳方法，并设计出最佳的外科植入方法。因此，这项提议的中心目标是利用一种综合的多学科多机构方法来生成、功能鉴定和随后的外科手术植入利用脱细胞肺中的干细胞产生的可存活的肺组织。这将通过以下两个具体目标来实现：1)优化胚胎和/或诱导的多能干细胞在去细胞小鼠肺中生长的条件，以获得具有功能的三维肺组织。2)为成功的体内移植和再细胞化肺的功能评估制定策略。 公共卫生相关性：永远不会有足够的供体肺来满足当前和未来的移植需求。相比之下，整个身体肺的去细胞化将产生一个完整的支架，可以用来自个别患者的干细胞(IPS)重新细胞化，然后用于自体移植。我们的初步数据表明，去细胞的小鼠肺可以用成体干细胞和胚胎干细胞重新细胞化。拟议的多机构合作研究将利用最先进的生物工程技术来开发在去细胞肺(包括身体肺)中生长功能肺组织的最佳方法，并设计出通过外科手术植入重新细胞肺的最佳方法。这将潜在地提供无限的无关捐赠者身体肺的供应，用于肺气肿、肺纤维化等疾病，以及目前尚无有效治疗方法的其他疾病。

DESCRIPTION (provided by applicant): There will never be enough donor lungs available to meet current and future transplantation needs. In contrast, de-cellularization of whole cadaveric lungs will result in an intact scaffold that can be re-cellularized with embryonic stem cells or with adult stem cells, including induced pluripotent stem cells (iPS) derived from individual patients and subsequently utilized for autologous transplantation. Notably, the de-cellularization process removes cellular antigens responsible for immune rejection and the de-cellularized lungs maintain native airway and alveolar architecture, extracellular matrix protein composition, and pulmonary vascular network. This will provide a potentially limitless supply of unrelated donor cadaveric lungs for use in diseases such as emphysema and other diseases for which there is currently no effective cure. Our preliminary data demonstrates that de-cellularized mouse lungs can be re-cellularized with both adult and embryonic stem cells. Further, the de-cellularized lung preparations can be assessed by both histologic and functional outcomes including lung mechanics, surfactant production, vascular perfusion, and epithelial barrier function, as the lungs re-cellularize. Most importantly, preliminary data demonstrates that the lungs can be surgically re-implanted in animal models. These data provide a firm platform for the proposed multi-institutional collaborative studies in which state-of-the-art bioengineering techniques will be utilized to develop optimal methods for growing functional lung tissue in de-cellularized lungs, including cadaveric human lungs, and to devise optimal approaches for surgical implantation. The central goal of this proposal is therefore to utilize a comprehensive multidisciplinary multi- institutional approach for generation, functional characterization, and subsequent surgical implantation of viable lung tissue produced using stem cells in the de-cellularized lungs. This will be accomplished in the following two Specific Aims: 1) To optimize conditions for development of functional three dimensional lung tissue from embryonic and/or induced pluripotent stem cells grown in de-cellularized mouse lungs. 2) To develop strategies for successful in vivo implantation and functional assessment of re- cellularized lungs. PUBLIC HEALTH RELEVANCE: There will never be enough donor lungs available to meet current and future transplantation needs. In contrast, de-cellularization of whole cadaveric lungs will result in an intact scaffold that can be re-cellularized with stem cells (iPS) derived from individual patients and subsequently utilized for autologous transplantation. Our preliminary data demonstrates that de-cellularized mouse lungs can be re-cellularized with both adult and embryonic stem cells. The proposed multi-institutional collaborative studies will utilize state-of-the-art bioengineering techniques to develop optimal methods for growing functional lung tissue in de-cellularized lungs, including cadaveric human lungs, and to devise optimal approaches to surgically implant the re-cellularized lungs. This will provide a potentially limitless supply of unrelated donor cadaveric lungs for use in diseases such as emphysema, pulmonary fibrosis, and others for which there is currently no effective cure.