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.

该项目将开发一种全功能、可植入的人类唾液腺，用于患有 头颈部肿瘤放疗后口干/口干。尽管监管机构要求 接近人类的动物模型用于临床移植组织工程器官，大型动物模型则不适用 存在测试干细胞恢复唾液功能的方法，包括液体分泌和蛋白质生产 对消化和口腔健康都有帮助。啮齿动物模型虽然对测试产品设计很有用，但缺乏关键属性 需要把可植入的生物装置送到诊所。为了解决这些缺点，我们集合了 一个跨学科团队，包括休斯顿的Farach-Carson/Harrington团队，Passineau实验室 在匹兹堡开发了一种使用辐射猪模型的方法，该模型可以进行基因操作，并且 位于安娜堡的Lombaert实验室评估种植体集成。我们最近的研究表明，猪体内的人 免疫抑制模型耐受性良好为发展异种移植提供了前所未有的机会 唾液人干/祖细胞(HS/PC)移植研究的模型和避免开发的需要 猪与猪之间的自体移植。这项提议建立在我们令人兴奋的初步结果的基础上，以开发和描述一部小说 辐射免疫抑制小型猪模型检测移植HS/PC修复唾液的能力 分泌功能。我们假设这个模型将重现辐射人体的环境。 唾液床，并提供了有朝一日评估人类S/P细胞功能所需的信息 移植到患有口干症的患者体内。具体目标是：1)建立辐射 免疫抑制小型猪作为适宜的宿主动物评价其长期稳定性、生物相容性和归宿 含有包裹的HS/PC的基质改性透明质酸(HA)水凝胶材料；a)开发和 标准化优化策略以将唾液肿瘤组织原型放置在宿主腮腺床中；b)评估 脱细胞水凝胶植入腮腺后不同时间的稳定性 根据需要进行配方更改，以确保长期的生物整合；c)使用量化评分系统 评价微囊化HS/PC构建物移植到放射损伤后随时间的变化 唾液床包括生存能力，生长而不过度生长，宿主整合包括血管和神经，以及 分化为涎腺结构，表达腺泡、导管和肌上皮标志物。2)评估能力 移植组织以恢复唾液功能：a)展示从 恢复性基因治疗联合唾液移植治疗受照射小型猪的唾液肿瘤组织 肿瘤组织；b)通过评估水通道蛋白5在肿瘤组织和液体中的表达来显示液体流动的恢复 使用直接唾液采集技术进行生产。唾液功能恢复的评估将得到帮助 通过使用基因修饰来创造分泌的Met-Luc S/P细胞，当被植入时将提供一个非侵入性的 只需采集宿主血液或唾液中的LUC样本，即可跟踪移植物随时间存活的情况。

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.