Collaborative Research: RECODE: On-line Feedback Control of Human Mesenchymal Stem Cell Chondrogenesis

合作研究：RECODE：人类间充质干细胞软骨形成的在线反馈控制

• 批准号: 2225559
• 负责人: Lawrence Bonassar
• 金额: $ 30万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2225559&HistoricalAwards=false
• 关键词: Collaborative; Research; RECODE; line; Feedback

Regenerating healthy cartilage from stem cells is challenging. In this Reproducible Cells and Organoids via Directed-Differentiation Encoding (RECODE) project a team of researchers from Washington State University and Cornell University aim to provide a new tissue engineering strategy to make cartilage. Adult stem cells will be stimulated to become mature chondrocytes, the cells that make cartilage, to produce the type of tissue needed for repairing damaged cartilage. The project's approaches seek to prevent cells from transitioning to osteocytes, the cells that make bone. This will be done by using small molecules that interfere with the genes that control the production of bone. Real time imaging of fluorescing molecules will monitor whether the cells are likely to make cartilage or bone. Feedback is used to change the cell growth conditions to keep cells within an ideal range. The properties of the tissues created will be tested for their flexibility and composition. Both Washington State and Cornell will work with minority participation programs to include underrepresented students in the research.This RECODE project aims to enhance tissue engineering by driving mesenchymal stem cells (MSCs) toward a stable chondrogenic lineage. The goal is to move tissue constructs in the direction of a functionally organized articular cartilage extracellular matrix (ECM) and prevent osteogenesis. It is hypothesized that cell proliferation and differentiation can be controlled by feedback bioprocess shear stresses, growth factor concentration, oscillating hydrostatic pressure, and gene silencing. The team will interrogate MSC maturation with a fiber optic bundle to sense, in real-time, co-upregulation of Sox9 mRNA, critically involved in orchestrating chondrogenesis, and Runx2 mRNA, which if overexpressed will promote unwanted progression toward hypertrophic calcification and ossification. Process variables will be tied to the cellular mRNA architecture and ECM manufacture through an experimental design model that will further refine feedback control rules. Spatial and temporal validation will be performed through bulk and single cell mRNA transcriptomics, and confocal strain and Fourier-transform infrared spectroscopy tissue mapping. The team will leverage Research Experience for Undergraduates (REU) support and partnerships with Louis Stokes Alliance for Minority Participation programs at Washington State University and Cornell University to recruit underrepresented students with the goal to see these students transition into tissue engineering related PhD programs.This RECODE project is jointly funded by the Engineering Biology and Health Cluster in the Division of Chemical, Bioengineering, Environmental, and Transport Systems and the Biomechanics and Mechanobiology Program in the Division of Civil, Mechanical, and Manufacturing Innovation.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

从干细胞再生健康的软骨是具有挑战性的。在这个通过定向分化编码(Recode)可复制的细胞和器官(Recode)项目中，来自华盛顿州立大学和康奈尔大学的一组研究人员旨在提供一种新的组织工程策略来制造软骨。成人干细胞将被刺激成为成熟的软骨细胞，即形成软骨的细胞，以产生修复受损软骨所需的组织类型。该项目的方法试图阻止细胞向骨细胞过渡，骨细胞是构成骨骼的细胞。这将通过使用小分子来实现，这些小分子干扰控制骨骼产生的基因。荧光分子的实时成像将监测细胞是否可能生成软骨或骨骼。反馈用于改变细胞的生长条件，使细胞保持在理想的范围内。将对所创建的组织的属性进行柔韧性和组成测试。华盛顿州和康奈尔大学都将与少数族裔参与计划合作，将代表人数不足的学生纳入研究。这项RECODE项目旨在通过推动间充质干细胞(MSCs)向稳定的软骨化谱系发展来加强组织工程。其目标是将组织结构朝着功能组织的关节软骨细胞外基质(ECM)的方向移动，并防止成骨。假设细胞的增殖和分化可以通过反馈的生物过程剪切力、生长因子浓度、振荡静水压力和基因沉默来控制。该团队将用光纤束询问MSC的成熟情况，以实时检测与软骨形成密切相关的Sox9和Runx2基因的共同上调，如果过度表达，将促进不必要的肥厚性钙化和骨化进展。过程变量将通过实验设计模型与细胞mRNA体系结构和ECM制造联系在一起，该模型将进一步完善反馈控制规则。空间和时间验证将通过批量和单细胞mRNA转录，以及共聚焦应变和傅里叶变换红外光谱组织图谱进行。该团队将利用本科生研究经验(REU)的支持以及与华盛顿州立大学和康奈尔大学的路易斯·斯托克斯少数民族参与联盟项目的合作伙伴关系来招募代表不足的学生，目标是看到这些学生过渡到与组织工程相关的博士课程。该RECODE项目由化学、生物工程、环境和运输系统部门的工程生物学和健康集群以及土木、机械和制造创新部门的生物力学和机械生物学项目联合资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。