Evaluation of pigment epithelium derived factor and its derived peptide for bone healing

色素上皮衍生因子及其衍生肽对骨愈合的评价

• 批准号: 10264787
• 负责人: CHRISTOPHER NIYIBIZI
• 金额: $ 17.3万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10264787
• 关键词: Acceleration; Adult; Affinity; Allografting; Amino Acids; Angiogenesis Inhibitors; Animal Model; Autologous Transplantation; BMP2 gene; BMP7 gene; Biomechanics; Bone Morphogenetic Proteins; Bone neoplasms; Cells; Cephalic; Clinical; Data; Defect; Development; Diagnostic radiologic examination; Evaluation; Face; Femoral Fractures; Femur; Forteo; Fracture; Fracture Healing; Genes; Greater sac of peritoneum; Harvest; Histologic; Histology; Impaired healing; In Vitro; Individual; Injections; Knock-out; Knockout Mice; Lead; Mesenchymal Stem Cells; Methods; Minerals; Morbidity - disease rate; Mus; Mutation; Natural regeneration; Osteoblasts; Osteocytes; Osteogenesis; Osteogenesis Imperfecta; Osteoid; Pathology; Patients; Peptides; Protocols documentation; Reporting; Saline; Site; Testing; Tissue Harvesting; Tissues; Trauma; Wild Type Mouse; base; bone; bone healing; disease transmission; fetal; healing; microCT; mineralization; mouse model; novel; novel therapeutic intervention; osteoblast differentiation; pigment epithelium-derived factor; pigment epithelium-derived factor receptor; stem cell differentiation

Abstract The present exploratory application proposes to evaluate the use of two novel factors, Pigment epithelium derived factor (PEDF) and a 17 amino acid peptide derived from it (P17) for the acceleration of healing femur fractures created in serpinf1 knockout mice or wildtype mice. PEDF encoded by serpinf1 gene is synthesized by a wide variety of cells in fetal and adult tissues. Mutations in serpinf1 gene cause type VI osteogenesis imperfecta (OI) whose hallmark is excessive osteoid build up that fails to mineralize. Bone defects resulting from trauma, bone tumors, and cranial facial defects may heal but a large number of them result in delayed healing or nonunion. Autografts are usually used but there is a limited supply and tissue harvest may lead to donor site morbidity. Allografts can be used but there is potential for disease transmission. Bone morphogenetic proteins (BMPs) specifically, BMP-2 and BMP-7 have been approved for clinical use to augment bone fracture healing. BMPs are very expensive, their mode of application remains problematic and they are prone to inducing ectopic bone formation. We and others reported that PEDF promoted mesenchymal stem cell (MSCs) differentiation and increased osteoblast mineralization and reduced expression of Sost/Sclerostin by osteocytes. Preliminary data showed that P17, a peptide with affinity for PEDF-receptor promoted osteoblast differentiation, increased matrix mineralization in vitro, and suppressed Sost expression by osteocytes. P17 infused into serpinf1 knockout mice via the peritoneal cavity, increased stiffness of the femurs which were reported to possess reduced stiffness. We are thus proposing to explore whether PEDF or P17 possess ability to accelerate healing of delayed or nonunion bone fractures using two mice models. The global hypothesize is PEDF or the P17 peptide will accelerate healing of critical sized mouse femoral defects created in serpinf1 knockout mice or wildtype mice. PEDF possess antiangiogenic activity but the P17 peptide does not, the peptide may possess superior ability to accelerate healing and regeneration of murine femoral fractures. We will test the hypothesis through two aims. In aim 1, we will determine whether P17 or PEDF delivered via the peritoneal cavity will accelerate healing of femur fractures created in serpinf1 knockout mice. Critical sized fractures will be created in serpinf1 mice femurs; the mice will be injected with P17 or PEDF via peritoneal cavity. Control mice will be injected with saline. Some mice will be followed by radiographic analysis at weekly intervals to determine the extent of healing. At week 6, all mice will be sacrificed, femurs harvested and evaluated by histology, micro CT and biomechanical testing. In aim 2, similar approaches will be carried out but using wildtype mice. We predict that Both P17 and PEDF will accelerate healing of femur fractures in both mice models but P17 may be more effective because it does not possess antiangiogenic activity as PEDF. Completion of the exploratory studies will lead to the development of novel therapeutic approaches to treat bone fractures as well as other bone pathologies based on PEDF or PEDF-derived peptide or peptides. .

抽象的 目前的探索性应用建议评估两个新因素的使用：色素上皮 衍生因子（PEDF）和17个氨基酸肽（p17），用于加速股骨 在Serpinf1敲除小鼠或野生型小鼠中产生的裂缝。由Serpinf1基因编码的PEDF合成 通过胎儿和成人组织中的各种细胞。 SERPINF1基因的突变导致VI型成骨发生 Imperfecta（OI）的标志过多的骨质骨质积聚，无法矿化。导致骨缺损 由于创伤，骨肿瘤和颅面缺陷可能会愈合，但其中大量导致延迟 康复或骨不连。通常使用自体移植，但供应有限，组织收获可能会导致 供体现场发病率。可以使用同种异体移植物，但可能会传播疾病。骨 形态发生蛋白（BMP）具体，BMP-2和BMP-7已被批准用于临床用途 增强骨折愈合。 BMP非常昂贵，其应用方式仍然有问题，并且 它们容易诱导异位骨形成。我们和其他人报告说PEDF促进了间充质 干细胞（MSC）分化和成骨细胞矿化的增加，并降低 骨细胞的刺激性/硬化蛋白。初步数据表明，P17，一种对PEDF受体亲和力的肽 促进了成骨细胞分化，体外基质矿化增加并抑制了SOST表达 由骨细胞。 P17通过腹膜腔注入Serpinf1敲除小鼠，增加了 据报道具有降低刚度的股骨。因此，我们建议探索PEDF还是 P17具有使用两种小鼠模型加速延迟或骨不连骨骨折的愈合的能力。这 全球假设是PEDF或P17肽将加速关键大小的小鼠股骨缺陷的愈合 用Serpinf1敲除小鼠或野生型小鼠创建。 PEDF具有抗血管生成活性，但p17肽 不，该肽可能具有较高的股毛发愈合和再生的能力 断裂。我们将通过两个目标检验假设。在AIM 1中，我们将确定是P17还是PEDF 通过腹膜腔传递的将加速在Serpinf1敲除小鼠中产生的股骨骨折的愈合。 将在股骨serpinf1小鼠中产生关键尺寸的裂缝；小鼠将通过P17或PEDF通过 腹膜腔。对照小鼠将注射盐水。一些小鼠将进行射线照相分析 每周间隔以确定愈合程度。在第6周，所有小鼠将被牺牲，股骨收获 并通过组织学，微CT和生物力学测试进行评估。在AIM 2中，将采用类似的方法 外出，但使用野生型小鼠。我们预测P17和PEDF都会加速股骨骨折的愈合 两种小鼠模型，但p17可能更有效，因为它不具有抗血管生成活性 佩夫。探索性研究的完成将导致新的治疗方法发展 根据PEDF或PEDF衍生的肽或肽治疗骨骨折以及其他骨骼病理。 。