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

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

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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（由适用提供）：该项目的长期目标是确定颌骨（ONJ）骨坏死的病理机制并发展预防和治疗方式。 ONJ是一些接受含氮双膦酸盐（BPS）治疗的患者的口服并发症经历，最近使用人源化的抗RANKL抗体。确认的临床ONJ病例的特征是通过口服粘膜异常伤口愈合制备的部分坏死的颚骨长期暴露。与公认的对破骨细胞的制药作用不同，BP在口服粘膜上的作用尚未完全阐明。许多研究表明，BP会影响肠溃疡的愈合，腹膜炎的发展以及激活的激活外围血液先天并获得免疫力。在快速将BP吸附到骨头或极端之前，给药路线（分别为PO，IP和IV）允许BP直接暴露于这些时间。但是，尚未确定影响口服粘膜的“自由” BP的来源。为了阐明与ONJ相关的口服粘膜异常的机制，我们假设可以通过破骨细胞移除jawbone预吸收的BP，并瞬时释放到口腔粘膜组织。我们设想，释放的BP的量可能取决于位于肺泡骨接口粘膜表面的破骨细胞生成程度。在达到临界浓度后，释放的BP可能直接影响口服粘膜居民细胞，并对口腔粘膜伤口的愈合产生不利影响。 To address this hypothesis, this exploratory R21 project proposals 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 problems will generate a fluorescent signal only in response to osteoclast-derived cathepsin K (CatK) activity which causes separation of emitter and quencher.由短的CATK可裂解的肽组成的新分子问题将通过独特的BP-Catk-Fret探针连接到生物活性或无效的BP脚手架上，将其连接到骨头或合成钙的磷酸盐，但将catk的信号catk emence catect和quencherative quencheration ementation和Activation。通过合成磷灰石涂层板和小鼠破骨细胞（将荧光信号与分辨率坑区域，Rap1a前丙基化和骨Osteoplast proptosis相关联），将在体外表征破骨细胞的BP-CATK-FRET激活。提出的问题还将是用于体内评估的强大工具，以建立由破骨细胞和口服粘膜居民细胞动员的BP之间的关键联系。它们也将有助于评估传统BP药物在确定ONJ风险和检查未来替代疗法方法中的作用。在实施该项目时，McKenna（USC）和Nishimura（UCLA）的研究团队将利用其在BP探针设计和化学（USC）（USC）和骨骼生物学模型（UCLA）方面的完整优势。