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. .

摘要 目前的探索性应用提出了评估两个新的因素，色素上皮细胞， 衍生因子（PEDF）和由其衍生的17个氨基酸的肽（P17），用于加速股骨愈合 在SerpINF 1敲除小鼠或野生型小鼠中产生的骨折。合成由serpinf 1基因编码的PEDF 由胎儿和成人组织中的多种细胞所控制。Serpinf 1基因突变导致VI型成骨 骨质疏松症（OI），其标志是类骨质过度堆积，无法矿化。骨缺损导致 创伤、骨肿瘤和颅面缺损可能会愈合，但其中大部分会导致延迟愈合。 愈合或不愈合。通常使用自体移植物，但供应有限，组织采集可能会导致 供区发病率。可以使用同种异体移植物，但有可能传播疾病。骨 形态发生蛋白（BMP），特别是BMP-2和BMP-7已被批准用于临床， 促进骨折愈合。BMP非常昂贵，其应用模式仍然存在问题， 它们易于诱导异位骨形成。我们和其他人报道了PEDF促进间充质干细胞增殖， 干细胞（MSC）分化和增加成骨细胞矿化和减少表达 骨细胞的Sost/Sclerostin。初步数据显示，P17，一种与PEDF受体具有亲和力的肽， 促进成骨细胞分化，增加体外基质矿化，抑制Sost表达 由骨细胞。通过腹膜腔将P17注入serpinf 1敲除小鼠，增加了 据报告，股骨刚度降低。因此，我们建议探讨是否PEDF或 使用两种小鼠模型，P17具有加速延迟或不愈合骨折愈合的能力。的 总体假设是PEDF或P17肽将加速临界尺寸小鼠股骨缺损的愈合 在Serpinf 1敲除小鼠或野生型小鼠中产生。PEDF具有抗血管生成活性，但P17肽 没有，该肽可能具有上级能力，加速小鼠股骨的愈合和再生。 骨折我们将通过两个目标来检验这个假设。在目标1中，我们将确定P17或PEDF 通过腹膜腔递送将加速SerpINF 1敲除小鼠中产生的股骨骨折的愈合。 将在serpinf 1小鼠股骨中产生临界尺寸的骨折;将通过皮下注射给小鼠注射P17或PEDF。 腹腔对照小鼠将注射生理盐水。一些小鼠将进行放射学分析 以确定愈合的程度。在第6周，处死所有小鼠，收获股骨 通过组织学、显微CT和生物力学测试进行评价。在目标2中，将采用类似的方法 但使用野生型小鼠。我们预测P17和PEDF都将加速股骨骨折的愈合， 这两种小鼠模型，但P17可能更有效，因为它不具有抗血管生成活性， PEDF。探索性研究的完成将导致开发新的治疗方法， 基于PEDF或PEDF衍生的肽或多种肽治疗骨折以及其它骨病变。 .