Notch-enriched Mesenchymal Stromal Cells for Bone Allograft Repair

富含缺口的间充质基质细胞用于同种异体骨移植修复

基本信息

批准号：
10269889
负责人：
Richard Shane Barton
金额：
$ 31.17万
依托单位：
LOUISIANA STATE UNIV HSC SHREVEPORT
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10269889
关键词：
Address Affect Allografting Animals Aspirate substance Autologous Transplantation Biological Assay Blood Vessels Bone Marrow Bone Tissue Bone callus Bromodeoxyuridine Cell Aging Cell Culture Techniques Cell Differentiation process Cell Lineage Cell Maintenance Cells Cellular Metabolic Process Characteristics Clinical Trials Data Defect Endothelial Cells Enzyme-Linked Immunosorbent Assay Exhibits Extracellular Matrix Femur Flow Cytometry Future Generations Genes Goals Human Immunohistochemistry Impairment Implant Label Ligands Mediating Mesenchymal Stem Cells Methodology Molecular Monitor Mus Mutagenesis Operative Surgical Procedures Osteogenesis Oxidative Stress Pathway interactions Patients Periosteum Population Process Recombinants Regulation Role Signal Transduction Site Skeletal Development Stains Stromal Cells TWIST1 gene Temperature Testing Therapeutic Tissue Engineering Transfection Transplantation United States Up-Regulation Vascularization Work allogenic bone transplantation angiogenesis base biomechanical test bone bone healing bone lead bone repair chromatin immunoprecipitation experimental study failure Implantation gain of function healing histological stains in vivo laser capture microdissection long bone loss of function mesenchymal stromal cell microCT mouse model next generation notch protein novel novel therapeutic intervention osteogenic pre-clinical promoter repaired skeletal stem cell proliferation stem cells stemness success tissue regeneration tissue repair transcription factor

项目摘要

Abstract: Segmental long bone defect surgeries require large devitalized allograft transplantations to replace missing host bone segments; however, significant problems often arise due to the impaired ability of the devitalized allograft to incorporate into the host bone. Recently, several discoveries in our lab have provided the basis for selecting, expanding and delivering of a substantial number of mesenchymal stem cells (MSCs), which retain their original stem cell characteristics, to be utilized in skeletal therapeutic applications: i) Notch2-selected and Notch activation by recombinant Jagged1 (JAG1) ligand expanded MSCs possess enhanced potentials for chondrogenic and osteogenic differentiation; ii) MSC sheets generated in temperature-responsive culture dishes exhibit a more robust effect on allograft incorporation into the host bone when transplanted to the site of the defect as a tissue-engineered periosteum surrounding the implanted graft; iii) Notch signaling induces rapid TWIST1 expression in MSCs. Based on these novel findings, we hypothesize that cell sheets generated with JAG1 expanded Notch2 positive MSCs on thermo-responsive culture dishes will significantly enhance critical segmented bone defects healing in a femoral allograft repair mouse model. To test this hypothesis, we will first to determine whether Notch2-selected and JAG1 expanded MSC sheets significantly enhance allograft healing and incorporation using our pre-clinical femoral allograft mouse model, as compared to cell sheets derived from traditionally isolated and expanded MSCs. Second, we will examine whether TWIST1 is require for JAG1-mediated MSC proliferation and maintenance in culture. Finally, we will determine how Notch activation by JAG1 involved in MSC sheet- induced angiogenesis. Data generated by this proposal will likely 1) identify optimized cell populations for generating tissue engineered periosteum (next generation MSC sheets), 2) validate a novel therapeutic approach for enhancing the repair of massive devitalized bone allograft and lead to advances in understanding the mechanism underlying MSC sheet-induced tissue repair, and 3) pave the path for large pre-clinical animal studies and subsequent clinical trials.

摘要：节段性长骨缺损手术需要大量失活的同种异体骨移植物移植以替代缺失的宿主骨段；然而，经常会出现重大问题由于失活的同种异体移植物融入宿主骨的能力受损而产生。近年来，我们实验室的多项发现为筛选、拓展和应用提供了基础。输送大量间充质干细胞 (MSC)，保留其原始状态干细胞特征，用于骨骼治疗应用：i) Notch2 选择和重组 Jagged1 (JAG1) 配体扩增的 MSC 的 Notch 激活具有增强软骨形成和成骨分化的潜力； ii) 生成的 MSC 表温度响应型培养皿对同种异体移植物掺入表现出更强的影响作为组织工程骨膜移植到缺损部位时的宿主骨围绕植入的移植物； iii) Notch 信号传导诱导 MSC 中快速 TWIST1 表达。基于这些新发现，我们假设 JAG1 生成的细胞片层扩展了热响应培养皿上的 Notch2 阳性 MSC 将显着增强关键能力股骨同种异体移植修复小鼠模型中分段骨缺损的愈合。为了检验这个假设，我们首先要确定Notch2选择和JAG1是否显着扩展了MSC片材使用我们的临床前股骨同种异体移植小鼠模型增强同种异体移植物的愈合和融合，与传统分离和扩增的 MSC 衍生的细胞片层相比。第二，我们将检查 JAG1 介导的 MSC 增殖和维持是否需要 TWIST1 在文化中。最后，我们将确定 JAG1 的 Notch 激活如何参与 MSC 表 - 诱导血管生成。该提案生成的数据可能会 1) 识别优化的单元格用于生成组织工程骨膜（下一代 MSC 片）的群体，2) 验证一种增强大量失活骨修复的新治疗方法同种异体移植并导致对 MSC 片诱导机制的理解取得进展组织修复，3）为大型临床前动物研究和后续临床铺平道路试验。