Matrix and Bioreactors for Human Lung Regeneration

描述（由申请人提供）：晚期或终末期肺部疾病，如肺纤维化、肺气肿和囊性纤维化，是美国发病率和死亡率的主要原因。尽管肺移植对一些患者来说是可行的替代方案，但大约50%的5年生存率（主要是由于慢性同种异体移植排斥和/或感染）限制了这种方式作为长期治疗。此外，供体肺的稀缺和移植后存活时间短是主要的限制。因此，迫切需要了解慢性肺部疾病的病理生理机制，以及设计创新的方法来修复或生物工程功能肺。因此，本研究将验证肺细胞外基质（ECM）驱动肺细胞的正确定位、分化和功能，是正常肺修复和再生的关键因素的假设。为了实现功能性人肺再生的长期目标，我们建议严格检查脱细胞肺外基质的组成，并开发评估肺基质结构和功能的新工具。此外，我们建议开发一种人肺生物反应器，在保持生理通气和灌注的同时，可以最佳地制备脱细胞的人肺基质。需要解决的问题包括：1)驱动细胞行为和功能的肺ECM的关键成分是什么？2)如何更好地评估肺ECM的屏障功能和结构？3)我们能否开发出一种生物反应器，在进行机械通气和血管灌注的同时，让人的肺进行脱细胞和再细胞化？这项U01应用的工作将有助于肺再生联盟的成功，因为我们将对用作肺生长底物的肺基质有一个基本和全面的了解。我们还将开发工具和生物反应器，使其他联盟成员能够利用和利用我们的发现。相关性：本研究的相关性是开始研究在实验室中再生功能性人肺的可行性。所提出的研究将全面定义正常人类肺的蛋白质组成，将设计新的方法来成像人类肺组织的细胞再生，并将开发一种新的生物反应器，在这种反应器中，人类肺可以再生。

DESCRIPTION (provided by applicant): Advanced or end-stage lung diseases, such as pulmonary fibrosis, emphysema, and cystic fibrosis, are a major cause of morbidity and mortality in the United States. Although lung transplantation is a viable alternative for some patients, the 5-year survival rate of approximately 50% (primarily due to chronic allograft rejection and/or infection) limits this modality as a long-term therapy. Moreover, the scarcity of donated lungs and the short graft viability times post-explant are major limitations. Thus, there i a desperate need both for understanding pathophysiologic mechanisms in chronic lung diseases as well as for devising innovative methods to repair or bioengineer functional lungs. Thus, this proposal will test the hypothesis that the lung extracellular matrix (ECM) drives the proper localization, differentiation, and function of lung cells and is the key contributor to norml lung repair and regeneration. With a long-term goal of regenerating a functional human lung, we propose to critically examine the constitutive makeup of the decellularized lung ECM and to develop new tools for assessing lung matrix structure and function. Further, we propose to develop a human lung bioreactor that allows for optimal preparation of decellularized human lung matrices while maintaining physiologic ventilation and perfusion. Among the questions to be addressed are: 1) what are the key components of lung ECM that drive cellular behavior and function? 2) How can we better assess the barrier function and structure of lung ECM? 3) Can we develop a bioreactor that will allow a human lung to be decellularized and subsequently recellularized while undergoing mechanical ventilation and vascular perfusion? Work in this U01 application will be instrumental for the success of the Lung Regeneration Consortium, because we will generate a basic and comprehensive understanding of the lung matrix that is used as a substrate for lung growth. We will also develop tools and bioreactors that will enable other Consortium members to utilize and leverage our findings. RELEVANCE: The relevance of this research is to begin studying the feasibility of regenerating a functional human lung in the laboratory. Studies proposed will comprehensively define the protein makeup of the normal human lung, will devise novel methods of imaging the cellular repopulation of human lung tissues, and will develop a novel bioreactor in which human lungs can be regenerated.