BRIGE: Delivery of Silencing RNA and TGF-b1 from Biopolymer Hydrogels to Inhibit Osteogenesis and Promote Chondrogenesis in Growth Plate Injuries

BRIGE：从生物聚合物水凝胶中传递沉默 RNA 和 TGF-b1，抑制生长板损伤中的成骨并促进软骨形成

• 批准号: 1342222
• 负责人: Melissa Krebs
• 金额: $ 17.5万
• 依托单位: Colorado School of Mines
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1342222&HistoricalAwards=false
• 关键词: BRIGE; Delivery; Silencing; RNA; TGF

Project Description: This research seeks to investigate a controlled, sustained delivery system for two different types of bioactive factors that could aid in the repair of injured growth plates in children. If the cartilage tissue in the growth plate is injured, bony tissue is deposited at the injured site, thus disrupting the growth plate and leading to abnormal bone growth. A new biomaterial delivery system that provides local, sustained delivery of silencing RNA and TGF-1 growth factor will be engineered, and the influence of the co-delivery of these bioactive factors on the chondrogenic differentiation of mesenchymal stem cells examined. Short interfering RNA (siRNA) provides a powerful gene silencing mechanism, inhibiting gene expression at the translational level by targeted destruction of specific mRNA sequences. Due to this ability to silence the expression of specific proteins, and since siRNA functions directly in the cytoplasm of the cell rather than being incorporated into the genomic DNA and its effect is transient, the impact of siRNA in clinical applications could be enormous. Unfortunately, the delivery of siRNA is currently a major hurdle to its clinical use, as siRNA is readily degraded and cleared from the body. The work in this proposal will provide new methods for delivering siRNA to a local site in a sustained, controlled manner. Co-delivery of growth factor to improve cartilage repair of growth plate injuries will also be examined. Thus, the engineering solution proposed here is the development of a delivery system capable of releasing two different bioactive factors, one to inhibit the formation of the bony tissue at the injury site and another to encourage the chondrogenic differentiation of stem cells that are present in the area so they form cartilage tissue. Broader Significance and Importance: Ultimately this technology could offer a much-needed therapeutic option for growth plate injuries in children. As children grow, their bones lengthen by mineralization of cartilage at the bone-cartilage interface in the growth plate. Injury to the growth plate can lead to the deposition of bony tissue within the cartilage, thus disrupting the growth plate. This disruption can result in serious consequences, including the bone not growing to its full length or the growth occurring abnormally such that the bone curves as it grows. In addition to addressing a clear clinical need, the results of the proposed work will provide new methods for localized and sustained delivery of silencing RNA, which could potentially benefit other applications in the future as well.Broader Impacts: This award will support both the increase and continuation of outreach efforts by Professor Krebs's lab. It will directly support the training of one graduate student for 2 years and the involvement of at least one undergraduate student and one high school intern in the project. The PI and graduate student will be participating in a summer camp for K-6 dyslexic students, designing science modules that will be presented to the students over the course of 2 of the 5 weeks of the summer camp. This will have a substantial impact on the students at the camp in awakening their scientific interest, and also on the PI and graduate student in learning more about dyslexia and ways that individuals can overcome it and/or use it to their benefit. The PI will also continue her support of hosting 1-2 female high school interns in the lab during the academic year through internship programs of local high schools. These students will work side-by-side with a graduate student and the PI on a specific project, and see it develop over the course of the year. Finally, the project would have broad societal impacts by providing a new biomaterial delivery system that could improve the treatment of growth plate injuries and also be used for other tissue regeneration applications in the future.This research has been funded through the Broadening Participation Research Initiation Grants in Engineering solicitation, which is part of the Broadening Participation in Engineering Program of the Engineering Education and Centers Division.

项目描述：本研究旨在调查两种不同类型的生物活性因素的受控，持续的分娩系统，这些因素可以帮助修复儿童受伤的生长板。如果生长板中的软骨组织受伤，则将骨组织沉积在受伤部位，从而破坏生长板并导致异常的骨骼生长。将设计一个新的生物材料输送系统，该系统将设计局部，持续的沉默RNA和TGF-1生长因子，并设计这些生物活性因子的共传递对所检查间充质干细胞软骨的软骨分化的影响。短干扰RNA（siRNA）提供了强大的基因沉默机制，通过针对特定的mRNA序列的靶向破坏来抑制转化水平的基因表达。由于这种能力使特定蛋白的表达保持沉默，并且由于siRNA直接在细胞的细胞质中起作用而不是被掺入基因组DNA中，其作用是短暂的，因此siRNA在临床应用中的影响可能是巨大的。不幸的是，siRNA的交付目前是其临床用途的主要障碍，因为siRNA很容易从体内退化并清除。该提案中的工作将提供新的方法，以持续，受控的方式将siRNA运送到当地地点。还将检查生长因子的共同传递以改善生长板损伤的软骨修复。因此，此处提出的工程解决方案是开发能够释放两个不同生物活性因子的输送系统，一种是抑制损伤部位骨组织的形成，另一个是为了鼓励在该区域中存在的干细胞的软骨分化，以形成软骨组织。更广泛的意义和重要性：最终，这项技术可以为儿童的生长板损伤提供急需的治疗选择。随着儿童的成长，他们的骨骼通过在生长板中的骨 - 果肉界面的软骨矿化而延长。生长板的损伤会导致软骨内骨组织的沉积，从而破坏生长板。这种破坏会导致严重的后果，包括骨头不长到其全长或异常生长，使骨曲线随着其生长而产生。除了满足明确的临床需求外，拟议的工作的结果还将为局部和持续交付沉默的RNA提供新的方法，这也可能在将来受益于其他应用。BROADER的影响：该奖项将支持Krebs教授实验室的外展工作的增加和延续。它将直接支持一名研究生2年的培训，并参与至少一名本科生和一名高中实习生。 PI和研究生将参加K-6阅读障碍学生的夏令营，设计科学模块，这些模块将在夏令营的5周内向学生展示给学生。这将对营地的学生产生重大影响，以唤醒他们的科学兴趣，以及对PI和研究生的了解更多有关阅读障碍的信息以及个人可以克服和/或使用它的方式的方式。 PI还将继续支持通过当地高中的实习计划在实验室内在实验室举办1-2位女性高中实习生。这些学生将与研究生和PI并排工作，并在一年中看到它的发展。最后，该项目将通过提供一种新的生物材料输送系统，可以改善生长板损伤的治疗，并在未来用于其他组织再生应用。这项研究是通过扩大的参与研究启动授予工程解决方案的研究来资助的，这是工程学实体的一部分。