Functional Biointegration of Bioengineered Salivary Tissues in Irradiated Animal Models

该项目的最终目标是为患者开发一种全功能、可植入的人类唾液腺。 头颈癌放射治疗后患口干症或慢性口干。 我们的团队最近开发了一种免疫抑制的、受辐射的人在迷你葡萄酒中的动物模型，用于临床前 翻译一种专利组织工程唾液组织替代物，我们称之为3D-ST。这么大 动物模型适用于测试基于细胞的项目，该项目旨在恢复唾液功能，包括 启动消化和维持口腔健康所需的水分分泌和蛋白质/酶的产生 空洞。作为补充，我们开发的辐射裸鼠模型是一个有用的测试模型 旨在最大化生物整合的产品设计，包括长期所需的血管和神经 器官成功。我们成功的跨学科团队包括UTHealth的Farach-Carson/Wu团队， 密歇根州的隆巴特团队和匹兹堡的斯威格外科团队。在三个地点运作，我们 开发了演示的实验成功工作流，该工作流利用了 每个站点。这项建议建立在我们令人兴奋的支持数据的基础上，使用辐照模型来演示 移植HS/PC在3D-ST中恢复唾液分泌功能的能力虽然我们到目前为止的工作 显示在3D-ST中植入的人/干祖细胞(HS/PC)具有长期(3-4个月)的活性 在免疫抑制的迷你葡萄酒和裸鼠中构建，一个重要的障碍仍然存在， 3D-ST与宿主种植床的永久生物整合。血管系统与唾液的生物整合 腺泡是提供营养和提供唾液的液体成分所必需的。稳定的神经支配对 腺体形态发生、细胞极性实现定向分泌和自主神经恢复 刺激分泌。这项建议建立在我们令人兴奋的支持数据的基础上，以鼓励移植人类 HS/PC重建唾液分泌功能，将重点从设计优化和植入转移到 使用我们的两个辐照模型进行成功的功能生物整合的可行性。我们假设两者都是 可以实现3D-ST的血管和周围神经整合，以稳定分化 唾液表型。这两个目标将：1)使用量化评分系统来评估生物集成 3D-STS植入受照动物模型的血管系统和自主神经系统 对唾液细胞表型的影响；2)评价移植的3D-ST修复唾液的能力 功能。这些目标的成功实现将改进现有的异种移植模型，用于测试 各种基于成人干细胞/祖细胞的治疗方法，以取代外分泌器官，并为首次非人类敞开大门 缓解口干症的试验。

The ultimate goal of this project is to develop a fully functional, implantable human salivary gland for patients suffering from xerostomia, or chronic dry mouth, subsequent to radiation therapy for head and neck cancer. Our team recently developed an immunosuppressed, irradiated human-in-miniswine animal model for preclinical translation of a patented tissue engineered salivary tissue replacement we call the 3D-ST. This large animal model is suitable for testing cell-based projects designed to restore salivary function that includes both water secretion and protein/enzyme production needed to initiate digestion and maintain health of the oral cavity. Complementing this, a radiated nude rat model we developed serves as a useful model for testing product designs purposed to maximize biointegration including both vasculature and nerve needed for longterm organ success. Our successful interdisciplinary team includes the Farach-Carson/Wu team at UTHealth, the Lombaert team in Michigan, and the Swegal surgical team in Pittsburgh. Functioning at three sites, we developed a demonstrated workflow for experimental success that takes advantage of the unique facilities at each site. This proposal builds on our exciting supporting data using irradiated models to demonstrate the ability of transplanted hS/PCs in the 3D-ST to restore salivary secretory function. While our work to date showed long term (3-4 months) viability of implanted human/stem progenitor cells (hS/PCs) in the 3D-ST construct in both immunosuppressed miniswine and nude rats, a significant hurdle remains to foster complete, permanent biointegration of the 3D-ST with the host implant bed. Biointegration of vasculature with salivary acini is needed to provide nutrition and to supply the fluid component of saliva. Stable innervation is critical for glandular morphogenesis, achievement of cell polarity for directional secretion, and restoration of autonomic stimulation of secretion. This proposal builds on our exciting supporting data to encourage transplanted human hS/PCs to reestablish salivary secretory function and moves our focus from design optimization and implant viability to successful functional biointegration using our two irradiated models. We hypothesize that both vasculature and peripheral nerve integration of the 3D-ST can be achieved to stabilize the differentiated salivary phenotype. The two aims will: 1) use a quantitative scoring system to evaluate biointegration of vasculature and autonomic nervous system into implanted 3D-STs in irradiated animal models and determine impact on salivary cell phenotype; and 2) evaluate the ability of the transplanted 3D-ST to restore salivary function. Successful achievement of these aims will improve existing xenotransplant models for testing a variety of adult stem/progenitor cell-based therapies to replace exocrine organs and open the door to first-inhuman trials for relief of xerostomia.