Engineered salivary gland tissue chips

工程唾液腺组织芯片

• 批准号: 10224168
• 负责人: Danielle S. Benoit
• 金额: $ 98.25万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224168
• 关键词: 3-Dimensional; Acinar Cell; Adhesives; Affect; Amylases; Benchmarking; Biochemical; Capsid Proteins; Cell Count; Cell Survival; Cell physiology; Cells; Characteristics; Chronic; Coupling; Crosslinker; Cues; DNA Damage; Data; Dependence; Development; Diagnosis; Duct (organ) structure; Ductal Epithelial Cell; Elastomers; Encapsulated; Endothelium; Engineering; Exhibits; Extracellular Matrix Proteins; FDA approved; Gel; Genetic; Gland; Goals; Head and Neck Cancer; Human; Hydrogels; Immune system; In Vitro; Label; Length; Libraries; Maintenance; Matrix Metalloproteinases; Microbubbles; Modeling; Molds; Mus; Nature; Nerve; Optics; Outcome; Paracrine Communication; Patients; Peptides; Pharmaceutical Preparations; Pharmacological Treatment; Phase; Phenotype; Plasma; Polymers; Prevention; Prolactin; Proteins; Radiation; Radiation Protection; Radiation Tolerance; Radiation induced damage; Radiation therapy; Radiation-Protective Agents; Resolution; Saliva; Salivary; Salivary Gland Tissue; Salivary Glands; Signal Transduction; Slice; Stains; Structure; Technology; Testing; Therapeutic; Tissue Microarray; Tissues; Xerostomia; autocrine; base; biomaterial compatibility; cholinergic; chromatin immunoprecipitation; density; drug efficacy; drug repurposing; ethylene glycol; head and neck cancer patient; high throughput screening; human tissue; improved; in vivo; irradiation; microphysiology system; mimetics; mouse model; paracrine; polydimethylsiloxane; preservation; regenerative; regenerative approach; screening; secretory protein; self assembly; stem; stem cells; tool

Abstract: For more than 550,000 patients annually diagnosed with head and neck cancers worldwide, severe loss of salivary gland function (xerostomia) is an unavoidable outcome of radiation therapy. There are presently no reliable and safe pharmacologic treatments for the resolution or prevention of radiation-induced xerostomia. Efforts to study radiosensitivity to discover effective radioprotective and regenerative strategies have been hampered by the inability to culture salivary gland mimetics in vitro, due to loss of secretory acinar cell phenotype. The principal milestone of this proposal is to engineer functional human salivary gland tissue chips to overcome this obstacle. Our labs have pioneered the use of hydrogel encapsulation to culture salivary gland cells in vitro. We have successfully demonstrated salivary gland cell survival up to 1 month post- encapsulation. Furthermore, cells organize into structures with apicobasal polarity and express secretory acinar markers, including Mist1. Although these data are promising, secretory marker expression is reduced compared to the native gland. Furthermore, the macroscale nature of hydrogels precludes their high- throughput use. Thus, we will utilize our microbubble (MB) array technology as a high-throughput, modular platform for the tissue chips. MBs are micron-scale spherical cavities molded in polydimethylsiloxane (PDMS). MBs have the advantage of length scales and curvatures similar to the secretory acinar unit of glands, providing a niche that promotes cell-cell contact and the concentration autocrine and paracrine factors that have been shown to enhance tissue assembly. Furthermore, MBs can be integrated with other microphysiological systems such as endothelial, nerve, and immune system chips. During the UG3 phase of this project, the go/no-go criteria will be the use of the MB platform to develop human gland tissue mimetics capable of long-term secretory function. Specifically for UG3, Aim 1 will use genetically labeled mouse acinar and duct cells to identify culture characteristics that maximize gland tissue mimetic function. Acinar and duct cell seeding ratios and densities will be varied in ‘blank’, extracellular matrix protein-functionalized, and in hydrogels all within MBs. Aim 2 (UG3) will validate the ability of human salivary gland cells to cellularly organize and maintain function in our previously developed macrogels and in within hydrogels in MBs, similarly to mouse cells in Aim 1. Our goal is to demonstrate functional human gland mimetic development in MB arrays by end of UG3. If successful, the UH3 phase will investigate hydrogel microenvironmental cues to further promote gland mimetic organization and function. Finally, Aim 3 will demonstrate the utility of gland mimetics by screening FDA-approved drugs to identify effective radioprotective agents. These compounds will be retroductally injected into mice to validate radioprotective potential. Successful development of salivary tissue chips will be transformative; by enabling in vitro analysis of functional gland mimetics, our ability to pursue therapeutic strategies, radioprotective and regenerative, will be dramatically improved.

摘要：全世界每年有超过55万名被诊断为头颈癌的患者，严重的 唾液腺功能丧失(口干症)是放射治疗不可避免的结果。确实有 目前还没有可靠和安全的药物治疗来解决或预防辐射引起的 口干燥症。努力研究辐射敏感性，以发现有效的辐射防护和再生策略 由于分泌腺泡的丧失，无法在体外培养唾液腺模拟物 细胞表型。这项提案的主要里程碑是设计出具有功能性的人类唾液腺组织 克服这一障碍的芯片。我们的实验室已经率先使用水凝胶胶囊来培养唾液 体外培养的腺体细胞。我们成功地证明了唾液腺细胞在移植后1个月内存活。 封装。此外，细胞组织成具有根尖基极的结构，并表达分泌 腺泡标志物，包括Mist1。虽然这些数据是有希望的，但分泌标记的表达减少了 与本土腺体相比。此外，水凝胶的宏观性质排除了它们的高性能。 吞吐量使用率。因此，我们将利用我们的微泡(MB)阵列技术作为高通量、模块化 纸巾芯片的平台。MBS是在聚二甲基硅氧烷(PDMS)中模制的微米级球形空腔。 MBS具有类似于腺体分泌腺泡单位的长度比例和曲率的优点， 提供促进细胞间接触和浓缩自分泌和旁分泌因子的利基， 已被证明可以增强组织组装。此外，MBS还可以与其他 微生理系统，如内皮、神经和免疫系统芯片。在UG3阶段 在这个项目中，去/不去的标准将是利用MB平台来开发人类腺体组织模拟物 具有长期的分泌功能。针对UG3，AIM 1将使用基因标记的小鼠腺泡细胞 和导管细胞，以确定最大限度地发挥腺组织模拟功能的培养特性。腺泡和导管 细胞播种率和密度将在空白、细胞外基质蛋白功能化以及在 水凝胶都在MBS中。目的2(UG3)将验证人唾液腺细胞对细胞的能力 类似地，在我们之前开发的宏凝胶和MBS中的水凝胶中组织和维持功能 目标1中的小鼠细胞。我们的目标是在MB阵列中展示模拟人类腺体的功能发育 到UG3结束时。如果成功，UH3阶段将调查水凝胶微环境线索，以进一步 促进腺体模拟组织和功能。最后，目标3将演示腺体模拟的实用性。 通过筛选FDA批准的药物来确定有效的辐射防护剂。这些化合物将是 经导管逆行注射到小鼠体内，以验证其辐射防护能力。唾液组织的成功发育 芯片将是变革性的；通过使功能腺体模拟的体外分析成为可能，我们追求 放射防护和再生的治疗策略将得到极大的改进。