Overcoming Barriers to Bioengineering 3-D Human Lung

克服生物工程 3-D 人肺的障碍

• 批准号: 8478182
• 负责人: Angela Panoskaltsis-Mortari
• 金额: $ 70.56万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-05 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8478182
• 关键词: 3-Dimensional; Activins; Adult; Air; Alveolar; Autologous; Biology; Biomedical Engineering; Biomimetics; Bioreactors; Blood Vessels; Bone Marrow; Cell-Matrix Junction; Cells; Characteristics; Chronic; Clinic; Clinical Research; Coculture Techniques; Cues; Development; Edema; Endothelial Cells; Epithelial; Epithelial Cells; Evaluation; Extracellular Matrix; Family suidae; Fetal Lung; Fibroblast Growth Factor 2; Gases; Gel; Goals; Growth; Head; Heart; Hemorrhage; Human; Immunosuppression; Implant; Liquid substance; Lung; Lung Transplantation; Mechanics; Mesenchymal; Organ; Organ Donor; Patients; Pattern; Pericytes; Physiological; Physiology; Property; Proteins; Rattus; Research Personnel; Source; Staging; Stem cells; Stretching; Stromal Cells; Structure; Structure of parenchyma of lung; Support System; Thoracic Surgical Procedures; Trachea; Transplantation; Vascularization; abstracting; embryonic stem cell; fetal; fibroblast growth factor 10; human adult stem cell; induced pluripotent stem cell; interest; lung development; lung repair; mortality; progenitor; scaffold

DESCRIPTION (provided by applicant): The overall goal of this application is to define conditions that are conducive to generating transplantable, 3-D human lung. Major obstacles to lung transplantation include the paucity of donor lungs available, chronic rejection the requirement for lifelong immunosuppression and high mortality. A new source of transplantable lung tissue that will not be subject to rejection is needed. Using decellularized lung matrix (DLM) as the natural foundational scaffold, our central hypothesis is that building functional, transplantable lung requires a combination of geometry that allows for appropriate cell localization and biomimetic cues superimposed on cells as they align to drive functional integration and maturation. The PI of this application developed a "decellularized, ventilated lung bioreactor" system that supports the growth of fetal lung cells and is amenable to evaluation of the potential to rebuild the lung. In addition to the use of DLM, the novelty of this proposal entails the use of autologous cells with lung recellularization potential all from the same BM source, i.e. all cells are autologous to each other and have the potential to be used in translational clinical studies. Post natal/adult human iPS cells are cells that have been reprogrammed to have characteristics of embryonic stem cells. We have evidence that DLMs can be seeded with induced pluripotent stem cells (iPS cells) that give rise to alveolar type II-like cells. We wish to evaluate the use of autologous BM-derived mesenchymal stromal cells (MSCs) and iPS-derived endothelial cells (ECs) in completing the recellularization of DLM. A team of investigators representing a convergence of the fields of bioengineering, pulmonary biology, physiology, stem cells, vascular biology and thoracic surgery has been assembled. Our aims encompass the recellularization of DLM in a developmental stage-appropriate manner that results in correct cell patterning with tight junctional barriers, gas exchange and fluid clearance functional properties. We will take a stepwise approach to determine the contribution of matrix geometry, physiological stretch and vascularization on re-epithelialization and function of recellularized constructs using human autologous progenitors and compare to native lung. We reason that by comparing different conditions head-to-head and in combination, several fundamental "rules" for bioengineering an autologous functional lung will be defined. There are 3 Specific Aims in this proposal. In Aim 1 we will determine the contribution of matrix geometry and physiological mechanical stretch on the differentiation of human progenitor cells into pulmonary cells. In Aim 2, we will evaluate if recellularization of DLMs is best achieved with mesenchymal induction using BM-derived MSCs. Aim 3 will determine if prior endothelialization of the DLM vascular network potentiates subsequent epithelial progenitor cell attachment to DLM, differentiation and function. We will use BM-derived endothelial cells (ECs) infused through the vasculature access to re-endothelialize DLM. Since vessels are stabilized by pericytes that are of MSC origin, we will use human BM-derived MSCs to study if they can potentiate the attachment of ECs. (End of Abstract)

描述（由申请人提供）： 该应用的总体目标是定义有利于生成可移植的3-D人肺的条件。肺移植的主要障碍包括可用的供体肺部缺乏，慢性拒绝终身免疫抑制和高死亡率的要求。需要一种不受排斥反应的可移植肺组织的新来源。使用脱细胞肺基质（DLM）作为天然基础支架，我们的中心假设是，构建功能性的，可移植的肺需要几何形状的组合，从而使细胞定位和仿生提示在细胞上叠加在细胞上以驱动功能整合和成熟。该应用的PI开发了一种“脱细胞，通风的肺生物反应器”系统，该系统支持胎儿肺细胞的生长，并且可以评估重建肺的潜力。除了使用DLM外，该提案的新颖性还包括使用具有肺部质细胞电位的自体细胞全部来自相同的BM来源，即所有细胞都是彼此自体的，并且有可能在转化临床研究中使用。出生时/成年人类IPS细胞是已重编程以具有胚胎干细胞特征的细胞。我们有证据表明，DLM可以用诱导的多能干细胞（IPS细胞）接种，从而导致肺泡类型II型细胞。我们希望评估自体BM衍生的间充质基质细胞（MSC）和IPS衍生的内皮细胞（EC）的使用在完成DLM的覆盖层。一组研究人员组成了代表生物工程，肺部生物学，生理学，干细胞，血管生物学和胸腔手术领域的融合。我们的目标包括DLM以发育阶段适当的方式覆盖的覆盖，从而导致具有紧密的连接屏障，气体交换和流体清除功能性能的正确细胞模式。我们将采用逐步的方法来确定基质几何，生理拉伸和血管化对使用人自体祖细胞的再细胞化构建体重新上皮化和功能的贡献，并与天然肺进行比较。我们认为，通过比较不同的条件并结合起来，将定义几种生物工程的基本“规则”。该提案中有3个具体目标。在AIM 1中，我们将确定基质几何形状和生理机械拉伸对人祖细胞分化为肺细胞的贡献。在AIM 2中，我们将评估使用BM衍生的MSC通过间充质诱导最好地实现DLM的覆盖化。 AIM 3将确定DLM血管网络的事先内皮化是否会增强随后的上皮祖细胞的附着DLM，分化和功能。我们将使用通过脉管系统进入重新内皮化DLM的BM来源的内皮细胞（EC）。由于血管的稳定在MSC起源的周细胞中，我们将使用人类BM衍生的MSC研究是否可以增强EC的附着。 （抽象的结尾）