Cell-Based Therapy in Minipig Model of Radiation-Induced Xerostomia

小型猪辐射诱发口干症模型的细胞疗法

• 批准号: 10214978
• 负责人: MARY C FARACH-CARSON
• 金额: $ 40.45万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214978
• 关键词: Address; Allografting; Animal Disease Models; Animal Model; Animals; Artificial Organs; Autologous Transplantation; Beds; Biological; Biology; Blood; Cell Therapy; Cells; Clinic; Collection; Comparative Pathology; Cues; Data; Development; Device Designs; Devices; Digestion; Duct (organ) structure; Encapsulated; Ensure; Environment; Enzymes; Epithelial; Epithelium; Evaluation; Excision; Family suidae; Feasibility Studies; Fluids and Secretions; Formulation; Genetic; Gland; Goals; Graft Survival; Growth; Head and Neck Cancer; Homologous Transplantation; Human; Hydrogels; Immunosuppression; Implant; Laboratories; Liquid substance; Lupus; Miniature Swine; Modeling; Modification; Myoepithelial; Nerve; Nude Rats; Oral cavity; Oral health; Organ; Paper; Parotid Gland; Patients; Population; Positioning Attribute; Production; Proteins; Protocols documentation; Publishing; Radiation; Radiation therapy; Radiology Specialty; Research; Research Personnel; Resources; Rodent Model; Saliva; Salivary; Salivary Glands; Sampling; Sjogren' s Syndrome; Source; Standardization; Stem cell transplant; Structure; Study models; System; Techniques; Testing; Therapeutic; Time; Tissue Engineering; Tissue Transplantation; Tissues; Transplantation; Water; Work; Xenograft Model; Xerostomia; aquaporin 5; base; biomaterial compatibility; clinical translation; design; digital; facial transplantation; fluid flow; functional restoration; gene therapy; genetic manipulation; head and neck cancer patient; human stem cells; hyaluronate; immunosuppressed; improved; in vivo; man; new technology; novel; post-transplant; prototype; reconstruction; restoration; side effect; skills; stem; stem cells; success

This project will develop a fully functional, implantable human salivary gland for patients suffering from xerostomia/dry mouth after radiotherapy for head and neck cancer. Despite the regulatory requirement for a close-to-human animal model for clinical translation of tissue engineered organs, large animal models do not exist to test stem-cell approaches to restore salivary function including fluid secretion and protein production needed for digestion and oral health. Rodent models, while useful to test product designs, lack key attributes needed to deliver an implantable biological device to the clinic. To address these shortcomings, we assembled an interdisciplinary team that includes the Farach-Carson/Harrington team in Houston, the Passineau laboratory in Pittsburgh to develop an approach using a radiated pig model amenable to genetic manipulation, and the Lombaert laboratory in Ann Arbor to assess implant integration. Our recent demonstration that a human-in-pig model under immunosuppression is well tolerated presents an unprecedented opportunity to develop xenograft model for the salivary human stem/progenitor cell (hS/PC) transplantation studies and avoid the need to develop a pig-in-pig autograft. This proposal builds on our exciting preliminary results to develop and characterize a novel radiated immunosuppressed mini-pig model for testing the ability of transplanted hS/PCs to restore salivary secretory function. We hypothesize that this model will recapitulate the environment of the radiated human salivary bed, and provide the type of information needed to evaluate the function of human S/P cells one day to be transplanted into patients suffering from xerostomia. Specific Aims are: 1) Establish the radiated immunosuppressed minipig as a suitable host animal to evaluate the long term stability, biocompatibility and fate of matrix-modified hyaluronate (HA) hydrogel materials containing encapsulated hS/PCs; a) Develop and standardize optimized strategies to place the salivary neotissue prototype in the host parotid bed; b) Evaluate stability of implanted acellular hydrogel constructs at various times after placement in the radiated parotid and make formulational changes as needed to ensure long term biointegration; c) Use a quantitative scoring system to evaluate the fate over time of encapsulated hS/PC-loaded constructs after transplantation into the radiated salivary bed including viability, growth without overgrowth, host integration including vasculature and nerve, and differentiation into salivary structures expressing acinar, ductal and myoepithelial markers. 2) Evaluate the ability of the transplanted tissue to restore salivary function: a) Demonstrate functional protein delivery from the salivary neotissues into the irradiated minipig by combining restorative gene therapy with implanted salivary neotissues; b) Demonstrate restoration of fluid flow by evaluating aquaporin 5 expression in neotissues and fluid production using direct saliva collection techniques. The evaluation of salivary functional restoration will be aided by use of a genetic modification to create a secreted Met-Luc S/P cell that when implanted would provide a noninvasive means of following graft survival over time simply by sampling Luc in host blood or saliva.

该项目将为患有患者 放射治疗后颈部和颈部癌后口干/口干。尽管有监管要求 用于组织工程器官的临床翻译的近接到人类动物模型，大型动物模型不会 存在用于恢复唾液功能（包括液体分泌和蛋白质产生）的测试干细胞方法 消化和口腔健康所需。啮齿动物模型虽然可用于测试产品设计，但缺乏关键属性 需要向诊所提供可植入的生物装置。为了解决这些缺点，我们组装了 一个跨学科的团队，包括休斯敦的Farach-Carson/Harrington团队，Passineau实验室 在匹兹堡，使用辐射的猪模型开发一种方法， Ann Arbor的Lombaert实验室评估植入物整合。我们最近的演示是人类的猪 在免疫抑制下的模型非常容忍为开发异种移植的前所未有的机会 唾液人类干/祖细胞（HS/PC）移植研究的模型，并避免需要发展 猪中的自体移植。该提案以我们令人兴奋的初步结果为基础 辐射的免疫抑制微杆杆模型，用于测试移植的HS/PC恢复唾液的能力 分泌功能。我们假设该模型将概括辐射人的环境 唾液床，并提供一日评估人类S/P细胞功能所需的信息类型 被移植到患有小心脏的患者中。具体目的是：1）建立辐射 免疫抑制的小ip作为合适的宿主动物，以评估长期稳定性，生物相容性和命运 基质修饰的透明质酸（HA）水凝胶材料，其中包含封装的HS/PC； a）发展和 标准化优化的策略，将唾液新疗法原型放在宿主腮腺床上； b）评估 放置在辐射腮腺和 根据需要进行公式更改，以确保长期生物整合； c）使用定量评分系统 评估封装的HS/PC负载构造的命运，将其移植到辐射中 唾液床，包括生存能力，生长没有过度生长，宿主整合，包括脉管和神经，以及 分化为表达腺泡，导管和肌上皮标记的唾液结构。 2）评估能力 移植组织以恢复唾液功能：a）表现出功能性蛋白质的递送 通过将恢复性基因治疗与植入的唾液结合起来，唾液新疗法进入了辐照的小杂志 新疗请； b）通过评估Neotissues和流体中的水通道蛋白5的表达来证明流体流的恢复 使用直接唾液收集技术生产。唾液功能恢复的评估将得到帮助 通过使用遗传修饰来创建一个分泌的MET-LUC S/P细胞，植入后会提供无创的 仅通过在宿主血液或唾液中采样LUC来遵循移植物生存的手段。

项目成果

Bringing hydrogel-based craniofacial therapies to the clinic.

• DOI: 10.1016/j.actbio.2021.10.056
• 发表时间: 2022-01-15
• 期刊: ACTA BIOMATERIALIA
• 影响因子: 9.7
• 作者: Trubelja, Alen;Kasper, F. Kurtis;Farach-Carson, Mary C.;Harrington, Daniel A.
• 通讯作者: Harrington, Daniel A.

Systematic Analysis of Actively Transcribed Core Matrisome Genes Across Tissues and Cell Phenotypes.

• DOI: 10.1016/j.matbio.2022.06.003
• 发表时间: 2022-06
• 期刊: Matrix biology : journal of the International Society for Matrix Biology
• 作者: Tristen V. Tellman;Merve Dede;V. Aggarwal;Duncan Salmon;A. Naba;M. Farach-Carson

Immunosuppressed Miniswine as a Model for Testing Cell Therapy Success: Experience With Implants of Human Salivary Stem/Progenitor Cell Constructs.

• DOI: 10.3389/fmolb.2021.711602
• 发表时间: 2021
• 期刊: Frontiers in molecular biosciences
• 影响因子: 5
• 作者: Wu D;Lombaert IMA;DeLeon M;Pradhan-Bhatt S;Witt RL;Harrington DA;Trombetta MG;Passineau MJ;Farach-Carson MC
• 通讯作者: Farach-Carson MC

MUC1 and Polarity Markers INADL and SCRIB Identify Salivary Ductal Cells.

MUC1 和极性标记 INADL 和 SCRIB 识别唾液管细胞。

• DOI: 10.1177/00220345221076122
• 发表时间: 2022
• 期刊: Journal of dental research
• 影响因子: 7.6
• 作者: Wu,D;Chapela,PJ;Barrows,CML;Harrington,DA;Carson,DD;Witt,RL;Mohyuddin,NG;Pradhan-Bhatt,S;Farach-Carson,MC
• 通讯作者: Farach-Carson,MC

Microfluidic coaxial 3D bioprinting of cell-laden microfibers and microtubes for salivary gland tissue engineering

用于唾液腺组织工程的载有细胞的微纤维和微管的微流控同轴 3D 生物打印

• DOI: 10.1016/j.bioadv.2023.213588
• 发表时间: 2023
• 期刊: Biomaterials Advances
• 作者: Yin, Yu;Vázquez-Rosado, Ephraim J.;Wu, Danielle;Viswananthan, Vignesh;Farach, Andrew;Farach-Carson, Mary C.;Harrington, Daniel A.
• 通讯作者: Harrington, Daniel A.