The Use of Coacervate Technology as a New Drug Delivery System for Musculoskeleta

使用凝聚技术作为肌肉骨骼的新型药物输送系统

• 批准号: 9130004
• 负责人: Johnny Huard
• 金额: $ 17.85万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-04 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9130004
• 关键词: Acute; Age; Angiogenesis Inhibitors; Angiogenic Factor; Area; Arthritis; Autologous; Autologous Transplantation; BMP2 gene; BMP4; Biomedical Engineering; Biomimetics; Blood Circulation; Bone Regeneration; Bone Transplantation; Calvaria; Cell Culture Techniques; Cell Therapy; Childhood; Chondrocytes; Clinical; Craniofacial Abnormalities; Data; Defect; Degenerative polyarthritis; Development; Disease; Drug Delivery Systems; Elderly; Engineered Gene; Epidemic; Etiology; Fracture; Gene Delivery; Genetic; Genetic Engineering; Goals; Gold; Grant; Growth Factor; Harvest; Healed; Health; Heparin; Human; In Vitro; Injection of therapeutic agent; Injury; Iodoacetates; Joints; Knee Osteoarthritis; Knowledge; Lead; Mediating; Methods; Modality; Modeling; Modification; Morbidity - disease rate; Mus; Muscle; Muscle satellite cell; Musculoskeletal; Natural regeneration; Older Population; Outcome; Pathologic; Patients; Population; Process; Proteins; Reporting; Research Design; Site; Skeletal Muscle; Skeleton; Stem cells; Supporting Cell; Surgeon; System; Technology; Tissue Engineering; Tissues; Translating; Translations; Transplantation; United States; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factors; Viral; Woman; Work; angiogenesis; articular cartilage; base; bone; bone healing; bone morphogenetic protein 2; bone morphogenetic protein 4; cartilage repair; clinically relevant; disability; gene therapy; healing; high risk; implantation; improved; in vivo; injured; interest; long bone; men; novel; novel therapeutics; operation; osteochondral tissue; osteogenic; osteogenic protein; polycation; prevent; regenerative; repaired; research study; socioeconomics; standard of care; stem cell technology; success; therapeutic protein

DESCRIPTION (provided by applicant): Incomplete healing of critical size bone defects, including defects of the craniofacial skeleton, are common. Osteogenic proteins, including bone morphogenetic protein 2 and 4 (BMP2, BMP4), promote bone healing, but the proteins have short half-lives and are rapidly cleared by the bloodstream, which limits their utility. The goals f our previous work were to develop gene therapy and tissue engineering approaches to efficiently deliver osteogenic proteins and improve bone healing using muscle-derived stem cells (MDSCs). We have shown that murine and human MDSCs (hMDSCs) genetically engineered to express BMP2 or 4 could differentiate toward an osteogenic lineage and improve bone healing in calvarial and long bone defects. We also found that concomitant gene delivery of vascular endothelial growth factor (VEGF) improves bone healing after the implantation of BMP2 or 4 expressing MDSCs. Similarly, we have reported that MDSCs isolated from mouse and human skeletal muscle were also capable of chondrogenic differentiation and could be used to promote articular cartilage repair after acute injury (osteochondral defects) and disease (osteoarthritis [OA]), especially when genetically modified to express bone morphogenetic protein 4 (BMP4-MDSC). However, in contrast to bone, our findings also suggested that genetic modification of MDSCs to express both BMP4 and the angiogenic antagonist sFlt-1, could accelerate the AC repair capacity of the cells supporting the fact that blocking angiogenesis is beneficial for AC repair. Although we have made substantial progress in muscle stem cell based therapy for bone and AC healing over the past number of years, most of our previous work involved genetic modification of the stem cells prior to transplantation, a step that limits te clinical translation of the work. We therefore propose a new set of experiments which will aim to circumvent the necessity of using viral transduction via the use of a novel heparin-polycation coacervate delivery system capable of slowly releasing the required therapeutic proteins including BMP2, VEGF and/or sFlt-1 to promote bone and AC repair in conjunction with non- transduced MDSCs. In the first set of experiments we will utilize the heparin-polycation coacervate delivery system to deliver BMP2 and VEGF to enhance hMDSC mediated bone repair. The second set of experiment will aim to promote AC repair after the induction of OA utilizing the heparin-polycation coacervate delivery system to slowly release BMP2 and sFlt-1, in combination with hMDSCs, to enhance AC repair. The efficiency of bone and AC repair with the coacervate-hMDSCs technology will be compared to both hMDSC based gene therapy and hMDSC based free protein therapy (i.e. without the use of the heparin-polycation coacervate). This application outlines a new area of research designed to offer valuable clinically relevant approaches based on novel bioengineering concepts for the treatment of musculoskeletal tissues following injury and disease.

描述(由申请人提供)：临界大小的骨缺损，包括头面部骨骼的缺陷，愈合不全是很常见的。成骨蛋白，包括骨形态发生蛋白2和4(BMP2，BMP4)，可促进骨愈合，但半衰期短，可被血流迅速清除，限制了其应用。我们之前工作的目标是开发基因治疗和组织工程方法，以有效地传递成骨蛋白，并利用肌肉来源的干细胞(MDSCs)改善骨愈合。我们已经证明，通过基因工程表达BMP2或BMP4的小鼠和人类MDSCs(HMDSCs)可以分化为成骨细胞系，并促进颅骨和长骨缺损处的骨愈合。我们还发现，伴随着血管内皮生长因子(VEGF)的基因传递促进了表达BMP2或4的MDSCs植入后的骨愈合。同样，我们也报道了从小鼠和人骨骼肌分离的MDSCs也具有向软骨分化的能力，并可用于促进急性损伤(骨软骨缺损)和疾病(骨关节炎[OA])后的关节软骨修复，特别是当基因修饰表达骨形态发生蛋白4(BMP4-MDSC)时。然而，与骨相反，我们的发现也表明，对MDSCs进行基因改造以表达BMP4和血管生成拮抗剂sFlt-1，可以加速细胞的AC修复能力，支持阻断血管生成有利于AC修复的事实。尽管在过去的几年里，我们在骨骼和AC愈合的基于肌肉干细胞的治疗方面取得了实质性的进展，但我们之前的大部分工作都是在移植前对干细胞进行基因改造，这一步骤限制了这项工作的临床转化。因此，我们提出了一套新的实验，旨在通过使用一种新型的肝素-聚阳离子凝聚体递送系统来避免使用病毒转导的必要性，该系统能够缓慢释放所需的治疗蛋白，包括BMP2、VEGF和/或sFlt-1，以促进骨和AC修复与非转导的MDSCs。在第一组实验中，我们将利用肝素-聚阳离子凝聚体递送系统来传递BMP2和血管内皮生长因子，以增强hMDSC介导的骨修复。第二组实验旨在促进骨性关节炎诱导后的AC修复，利用肝素-聚阳离子凝聚体递送系统缓慢释放BMP2和sFlt-1，与hMDSCs结合，增强AC修复。使用凝聚-hMDSCs技术修复骨骼和AC的效率将与基于hMDSC的基因治疗和基于hMDSC的游离蛋白治疗(即不使用肝素-聚阳离子凝聚体)进行比较。这项申请概述了一个新的研究领域，旨在提供基于新的生物工程概念的有价值的临床相关方法，用于治疗损伤和疾病后的肌肉骨骼组织。

项目成果

The use of heparin/polycation coacervate sustain release system to compare the bone regenerative potentials of 5 BMPs using a critical sized calvarial bone defect model.

• DOI: 10.1016/j.biomaterials.2022.121708
• 发表时间: 2022-09
• 期刊: Biomaterials
• 影响因子: 14

Influence of fixation on CA4+ contrast enhanced microCT of articular cartilage and subsequent feasibility for histological evaluation.

• 发表时间: 2021
• 期刊: American journal of translational research
• 影响因子: 2.2
• 作者: Xueqin Gao;Amit N. Patwa;Zhenhan Deng;Hajime Utsunomiya;M. Grinstaff;Joseph J. Ruzbarsky;B. Snyder;S. Ravuri;M. Philippon;J. Huard