Molecular and cellular mechanism of ONJ related to osteoclast inhibition

ONJ抑制破骨细胞相关的分子细胞机制

基本信息

批准号：
8638613
负责人：
CHARLES E MCKENNA
金额：
$ 25.18万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8638613
关键词：
Acute-Phase Reaction Address Adsorption Affect Affinity Antibodies Apatites Apoptosis Area Biology Bone necrosis Buffers Cells Cessation of life Chemistry Clinical Complication Culture Media Dentures Development Energy Transfer Evaluation Excretory function Fluorescence Fluorescent Dyes Functional disorder Future Generations Generic Drugs Goals Healed In Vitro Inflammation Intestines Investigation Jaw Kinetics Laboratories Link Mandible Maxilla Mediating Modality Modeling Molecular Molecular Probes Monitor Mus Natural Immunity Necrosis Nitrogen Oral Oral mucous membrane structure Osteoclasts Outcome Study Pathogenesis Patients Peptides Peritonitis Pharmaceutical Preparations Prevention Preventive Property Reagent Replacement Therapy Reporter Research Research Project Grants Risedronate Risk Role Route Side Signal Transduction Site Source Stomach Structure Surface Surgical wound TNFSF11 gene Technology Testing Therapeutic Tissues Tooth Extraction Trauma Ulcer Venous Wound Healing Zoledronate acquired immunity alveolar bone aqueous base bisphosphonate bone calcium phosphate cathepsin K design drug mechanism experience flu healing in vitro Model in vivo novel oral irritation osteoclastogenesis peripheral blood prenylation public health relevance response scaffold sensor therapeutic target tool wound

项目摘要

DESCRIPTION (provided by applicant): The long-term objectives of this project are to determine the pathological mechanism of osteonecrosis of the jaw (ONJ) and to develop preventive and therapeutic modalities. ONJ is an oral complication experienced by some patients treated with nitrogen-containing bisphosphonates (BPs) and more recently with humanized anti- RANKL antibody. Confirmed clinical ONJ cases are characterized by the prolonged exposure of partially necrotic jawbone facilitated by abnormal wound healing of oral mucosa. Unlike the well-established pharmacological effect on osteoclasts, the role of BP on oral mucosa has not been fully elucidated. A number of studies have demonstrated that BP affects the healing of intestinal ulceration, the development of peritonitis, and the activation of peripheral blood innate and acquired immunity. The administration routes (PO, IP and IV, respectively) allow the direct exposure of BP to these tissues, prior to rapid adsorption of BP to bone or excretion. However, the source of "free" BP affecting the oral mucosa has not been identified. To elucidate the mechanism of oral mucosa abnormality associated with ONJ, we have hypothesized that BP pre-adsorbed on the jawbone can be removed by osteoclasts and transiently released to the oral mucosa tissue. We envision that the amount of released BP may depend on the degree of osteoclastogenesis localized at the surface of alveolar bone interfacing oral mucosa; and upon reaching a critical concentration, the released BP may directly affect oral mucosa resident cells and adversely influence the healing of oral mucosa wound. To address this hypothesis, this exploratory R21 project proposes the synthesis and application of novel BP-based probes capable of delivering a Förster resonance energy transfer (FRET)-quenched reporter function to osteoclasts, where the probes will generate a fluorescent signal only in response to osteoclast-derived cathepsin K (CatK) activity which causes separation of emitter and quencher. Novel molecular probes consisting of a short CatK- cleavable peptide bridging a fluorescent dye and a quencher will be attached to a biologically active or inactive BP scaffold through unique BP-linker chemistry developed at USC. The silent BP-CatK-FRET probe will be strongly adsorbed to bone or synthetic calcium phosphate, but osteoclastic CatK will remove the quencher and activate the fluorescent signal. BP-CatK-FRET activation by osteoclasts will be characterized in vitro using synthetic apatite-coated plates and mouse osteoclasts (correlating fluorescent signal with resorption pit area, Rap1a prenylation and osteoclast apoptosis). The proposed probes will also be powerful tools for in vivo evaluation to establish the critical link between BP mobilized by osteoclasts and oral mucosa resident cells. They will also be useful to assess the role of legacy BP drugs in determining ONJ risk and to examine future replacement therapy approaches. In implementing this project, the research teams of McKenna (USC) and Nishimura (UCLA) will leverage their complementary strengths in BP probe design and chemistry (USC) and expertise in bone biology models (UCLA).

描述（由申请人提供）：该项目的长期目标是确定颌骨坏死（ONJ）的病理机制并开发预防和治疗方法 ONJ 是一些接受过治疗的患者所经历的口腔并发症。含氮双磷酸盐 (BP) 以及最近的人源化抗 RANKL 抗体已确诊的临床 ONJ 病例的特点是异常导致的部分坏死颌骨长期暴露。与已明确的破骨细胞药理作用不同，BP 对口腔粘膜的作用尚未完全阐明，许多研究表明 BP 会影响肠道溃疡的愈合、腹膜炎的发生和发展。的激活外周血先天免疫和获得性免疫在 BP 快速吸附到骨骼或排泄之前，给药途径（分别为 PO、IP 和 IV）允许 BP 直接暴露于这些组织。为了阐明与 ONJ 相关的口腔粘膜异常的机制，我们开发了预吸附在颌骨上的 BP 可以被破骨细胞去除并短暂释放到口腔。我们预计，释放的 BP 量可能取决于位于口腔粘膜界面的牙槽骨表面的破骨细胞生成程度；达到临界浓度后，释放的 BP 可能会直接影响口腔粘膜驻留细胞，并对口腔粘膜组织产生不利影响。为了解决这一假设，这个探索性的 R21 项目提出了合成和应用能够提供福斯特共振能量转移的新型 BP 探针。 (FRET) 猝灭报告功能对破骨细胞的作用，其中探针仅响应破骨细胞衍生的组织蛋白酶 K (CatK) 活性而产生荧光信号，从而导致发射器和猝灭剂分离，由短 CatK 可裂解肽组成。桥接荧光染料和猝灭剂将通过南加州大学 The Silent 开发的独特 BP 连接化学连接到生物活性或非活性 BP 支架上。 BP-CatK-FRET 探针将强烈吸附在骨或合成磷酸钙上，但破骨细胞 CatK 将去除猝灭剂并激活荧光信号。破骨细胞对 BP-CatK-FRET 的激活作用将在体外使用合成磷灰石涂层板进行表征。小鼠破骨细胞（将荧光信号与吸收坑面积、Rap1a 异戊二烯化和破骨细胞凋亡相关联）也将成为体内评估建立的强大工具。破骨细胞动员的 BP 和口腔粘膜驻留细胞之间的关键联系也将有助于评估传统 BP 药物在确定 ONJ 风险中的作用，并检查未来的替代治疗方法。 USC）和 Nishimura（加州大学洛杉矶分校）将利用其在 BP 探针设计和化学（USC）方面的互补优势以及骨生物学模型（UCLA）方面的专业知识。