Dynamic 3D interplay of primary human salivary cells and the basement membrane

人类原代唾液细胞和基底膜的动态 3D 相互作用

• 批准号: 8783875
• 负责人: Danielle Wu
• 金额: $ 5.41万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8783875
• 关键词: Acinar Cell; Acinus organ component; Address; Basement membrane; Biological; Biological Models; Biomechanics; Cell Survival; Cell physiology; Cells; Collaborations; Computer Simulation; Connexins; Cues; Deposition; Development; Diagnosis; Encapsulated; Engineering; Evaluation; Excision; Exertion; Extracellular Matrix; Feedback; Fluorescence Microscopy; Frequencies; Future; Gel; Gland; Goals; Grant; Growth; Head and Neck Cancer; Health; Human; Hyaluronic Acid; Hydrogels; Image; In Vitro; Integrins; Intervention; Lead; Life; Maintenance; Manuscripts; Maps; Mastication; Measures; Mechanics; Mediating; Mentorship; Modeling; Molecular; Movement; Natural regeneration; Neurotransmitters; Nuclear; Oral health; Parotid Gland; Patients; Positioning Attribute; Process; Proteins; Quality of life; Radiation therapy; Rattus; Research; Research Personnel; Resected; Resolution; Risk; Role; Rotation; Salivary; Salivary Gland Tissue; Salivary Glands; Serous; Shapes; Signal Transduction; Signaling Molecule; Staging; Standardization; Structure; Surgical Replantation; Symptoms; System; Tissue Engineering; Tissues; Traction; Training; Translations; Writing; Xerostomia; base; cancer therapy; career; cell assembly; cell motility; cell type; driving force; extracellular; fluorescence imaging; head and neck cancer patient; in vitro Model; in vivo; irradiation; light microscopy; loss of function; migration; multidisciplinary; novel; polarized cell; response; salivary cell; scaffold; skills; time use; tumor

DESCRIPTION (provided by applicant): Head and neck cancer treatments that require the resection of glandular tissue in combination with irradiation therapy cause significant damage to target and surrounding tissues. Salivary glands are vulnerable and when compromised, a loss of function causes hyposalivation and 'dry mouth' (xerostomia) that lead to an increase in oral health risk and a decline in quality of life. Currently, there is no cure for xerostomia, only interventions for alleviating the discomfort associated with loss of salivary function. A novel therapy is underway to address this unmet need using hyaluronic acid (HA) hydrogels and primary cells from resected human salivary gland. Repeatable organization of salivary acini into functional secretory units is a key step toward the standardization of our model system. Our goal is to mimic the microenvironment of parotid salivary gland tissue in development to best support the organization of serous acini. Determining factors for organized acini are basement membrane (BM) deposition and lumen formation. Using live imaging light and fluorescence microscopy, we have observed the coordinated motility of acinar cells in HA hydrogels, prior to acini organization. In early stage organization, acinar cells are in a dynamic microenvironment continuously influenced by mechanical forces, and we hypothesize that mechanical forces drive the BM deposition, lumen formation, and structural integrity of the acini in 3D. In Aim 1, we intend to identify the signaling mechanisms involved in the net coordination of cell motility durin BM deposition and growth of the acini. Signaling mechanisms proposed in this coordination include integrin signaling at the cell-ECM interface, connexin mediated intra- and intercellular signaling, and nesprin4 nuclear repositioning. In Aim 2, we will measure the traction forces required to initiate the coordinated movement of a multicellular structure in 3D. Live-fluorescence imaging and computational modeling will be used to develop displacement field and cellular traction maps, and reconstruct cellular traction forces of organizing acini as a function of their size and microenvironment. In Aim 3, we will evaluate acini organization, lumen formation, and structural integrity in response to mechanical loading of the hydrogel. Effects of varying magnitude and frequency loads on acini organization and integrity will be evaluated. Successful completion of these specific aims will (1) standardize the model system used to engineer serous acini of the salivary parotid gland as well as the evaluation process for optimizing iterations of the HA hydrogel herein, (2) yield fundamental understanding of salivary acini structure/function relations, and (3) advance the translational potential of this tissue engineered system. Additionally, the PI, Dr. Danielle Wu, will gain experimental and computational training in 3D, crucial for her future as an independent researcher in tissue engineering, will acquire skills and perspective from operating at a multidisciplinary interface, and will advance her long-term career goals with training in manuscript and grant writing, mentorship, and collaboration skills.

描述（由申请人提供）：需要切除腺组织与辐照疗法结合切除的头颈癌治疗，对目标和周围组织造成了重大损害。唾液腺很脆弱，当受到损害时，功能损失会导致缺乏症和“干嘴”（静态），这会导致口腔健康风险增加和生活质量下降。目前，尚无治愈静态的治疗方法，仅是减轻与唾液功能丧失相关的不适的干预措施。正在进行一种新的疗法，可以使用透明质酸（HA）水凝胶和切除的人类唾液腺的原代细胞来解决这种未满足的需求。可重复的唾液acini将其重复组织为功能分泌单位是朝着模型系统标准化的关键步骤。我们的目标是模仿发育中腮腺唾液腺组织的微环境，以最大程度地支持浆液性刺激的组织。有组织的acini的确定因素是基底膜（BM）沉积和管腔形成。使用实时成像光和荧光显微镜，我们已经观察到在ACINI组织之前，HA水凝胶中腺泡细胞的协调运动性。在早期的组织中，腺泡细胞处于由机械力不断影响的动态微环境中，我们假设机械力驱动了ACINI在3D中的BM沉积，管腔形成和结构完整性。在AIM 1中，我们打算确定细胞运动净协调性杜林BM沉积和ACINI生长的信号传导机制。该协调中提出的信号传导机制包括在细胞ECM界面处的整联蛋白信号传导，连接蛋白介导的细胞内信号传导和NESPRIN4核重复作用。在AIM 2中，我们将测量启动3D多细胞结构协调运动所需的牵引力。实时荧光成像和计算建模将用于发展位移场和细胞牵引图，以及重建组织acini的细胞牵引力，作为其大小和微环境的函数。在AIM 3中，我们将根据水凝胶的机械负载来评估ACINI组织，管腔形成和结构完整性。将评估不同大小和频率负载对ACINI组织和完整性的影响。这些特定目的的成功完成将（1）标准化用于设计唾液腮腺的浆液性卵泡的模型系统，以及用于优化此处HA水凝胶迭代的评估过程，（2）对唾液ACINI结构/功能关系的基本了解，以及（3）推进此组织工程系统的转化潜力。此外，PI，Danielle Wu博士将在3D中获得实验和计算培训，这对于她作为组织工程领域的独立研究人员的未来至关重要，将通过在多学科界面中获得技能和观点，并将通过手稿，指导，导师和协作技能来促进她的长期职业目标。