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：
10543407
负责人：
Richard Shane Barton
金额：
$ 31.49万
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/10543407
关键词：
Address Affect Allografting Animals Autologous Transplantation Binding Biological Assay Blood Vessels Bone Marrow Bone Marrow Aspiration 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 Induction 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 biomechanical test bone bone healing bone lead bone marrow mesenchymal stem cell 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 transdifferentiation

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：节段性的长骨缺陷手术需要大量的转移同种异体移植移植以取代缺失的宿主骨骼段；但是，经常重大问题由于失去活力的同种异体移植物掺入宿主骨骼的能力受损而产生。最近，我们实验室中的几个发现为选择，扩展和提供大量的间充质干细胞（MSC），它们保留其原始干细胞特征，可用于骨骼治疗应用：i）Notch2选择和重组JAGGED1（JAG1）配体扩展MSC的Notch激活具有增强软骨和成骨分化的潜力； ii）在温度响应性培养皿对同种异体移植的融合具有更强的效果宿主骨作为组织工程骨膜移植到缺陷部位时围绕植入的移植物； iii）Notch信号传导在MSC中诱导快速扭曲1的表达。基于这些新颖的发现，我们假设用JAG1生成的细胞表扩展了热响应性培养皿上的Notch2阳性MSC将显着增强关键分段的骨缺损可在股骨同种异体修复小鼠模型中愈合。为了检验这一假设，我们将首先确定Notch2选择和JAG1是否大大扩展了MSC表使用我们的链式股骨同种异体移植小鼠模型增强同种异体移植愈合和掺入与传统上孤立和扩展的MSC得出的细胞表相比。第二，我们将检查twist1是否需要JAG1介导的MSC增殖和维护在文化中。最后，我们将确定如何参与MSC纸的JAG1激活Notch激活诱导血管生成。该提案生成的数据可能会1）确定优化的单元格生成组织工程骨膜的种群（下一代MSC表），2）验证一种新型的治疗方法，以增强大规模退还骨的修复同种异体移植并导致理解MSC纸张诱导的机制的进步组织修复和3）为大型临床前动物研究和随后的临床铺平道路试验。