HIF-1alpha Mediated Wound Healing in Periapical Lesion

HIF-1alpha 介导的根尖周病变伤口愈合

基本信息

批准号：
8692733
负责人：
HAJIME SASAKI
金额：
$ 24.64万
依托单位：
ADA FORSYTH INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-02 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8692733
关键词：
Abscess Acute Anti-Inflammatory Agents Anti-inflammatory Bone Regeneration Chronic Code Decontamination Dental Research Development Disease Elements Endodontics Enzyme-Linked Immunosorbent Assay Event Exhibits Failure Gene Expression Genes Goals Granuloma Granulomatous HIF1A gene Healed Histology Immunocompetent Immunohistochemistry Individual Infection Inflammation Inflammatory Injection of therapeutic agent Interleukin-10 Jaw Knock-out Knockout Mice Laboratories Lentivirus Vector Lesion Macrophage Activation Measures Mediating Modeling Molecular Mus Osteitis Outcome Pathogenesis Pathway interactions Phenotype Process Procollagen-Proline Dioxygenase Pulp Canals Refractory Reporting Resolution Retreatment Role Secure System Therapeutic Toll-Like Receptor 2 Tooth Extraction Tooth root structure Treatment outcome Up-Regulation Virus Wild Type Mouse Wound Healing YC-1 bone bone loss diabetic healing hypoxia inducible factor 1 improved in vivo inhibitor/antagonist innovation macrophage novel novel strategies novel therapeutic intervention novel therapeutics pathogen periapical public health relevance recombinant virus response seal small hairpin RNA treatment strategy

项目摘要

DESCRIPTION (provided by applicant): A periapical lesion is an infection-induced inflammation within the jaws that ultimately results in the destruction of bone that surrounds the apex of the tooth root. Periapical lesions are often refractory to treatment and require retreatments or, in worst case, tooth extraction. Thus, there is an acute need for novel therapeutic approaches that improve the healing of periapical lesions. The long-term goal of this project is to identify the molecular networks that regulate wound healing after infection-induced inflammation and bone destruction. Our preliminary studies showed that the Toll-like receptor 2 (TLR2)/Interleukin-10 (IL-10) double deficient (dKO) mouse infected with common endodontic pathogens exhibits an acute periapical lesion that subsequently exhibiting spontaneous wound healing principally bone regeneration The healing response present in dKO mice is strongly associated with up-regulation of HIF-1a (hypoxia inducible factor 1 alpha) expression in the lesion and sequential activation of proinflammatory M1 and anti-inflammatory M2 macrophages. These findings suggest that HIF-1a activation is a key element for improved periapical wound healing. The central hypothesis of this proposal is that HIF-1a activation leads to periapical wound healing via induction of M1 macrophage activation followed by emergence of M2 to facilitate final clearance of the lesion. Conversely, failure of HIF-1a activation results in failue of M1 activation followed by delayed and polarized M2 macrophage activation, which leads to chronic inflammation and radicular granuloma. Thus, the HIF-1a- mediated response is a target pathway for the development of novel therapeutics for improving periapical wound healing. The overall goal of this exploratory proposal is to determine whether HIF-1a activation will predictably induce periapical lesion in immunocompetent (WT) mice. The goal will be achieved through two specific aims. AIM 1 is to assess the impact of HIF-1a modulation on periapical lesion formation and healing in WT mice. Lentiviral vectors coding HIF-1a shRNA and constitutively active form of HIF-1a will be used to modulate HIF-1a activity. AIM 2 is to determine the role of endogenous HIF-1a on macrophage activation and phenotype in vivo. HIF-1a will be inactivated in dKO mice, and endogenous HIF-1a will activated by chemically inhibiting prolyl hydroxylase that is essential for HIF-1 activation in order to measure the effect on macrophage activation and the course infection. The proposed studies will provide new information about the fundamental role HIF-1a in periapical lesions, which is a novel wound healing pathway that involves M1/M2 sequential macrophage activation, and demonstrate a therapeutic potential of HIF-1a in dentoalveolar inflammation. These outcomes are targeted to novel treatment strategies to improve the healing process in dentoalveolar infections that will have potential application in other chronic fibrotic and granulomatous diseases.

描述（由申请人提供）：根尖周病变是颌骨内感染引起的炎症，最终导致牙根尖周围的骨质破坏。根尖周病变通常难以治疗，需要再治疗，或者在最坏的情况下需要拔牙。因此，迫切需要改善根尖周病变愈合的新治疗方法。该项目的长期目标是确定在感染引起的炎症和骨质破坏后调节伤口愈合的分子网络。我们的初步研究表明，感染常见牙髓病原体的 Toll 样受体 2 (TLR2)/白细胞介素 10 (IL-10) 双缺陷 (dKO) 小鼠表现出急性根尖周病变，随后表现出自发伤口愈合，主要是骨再生。 dKO 小鼠中存在的反应与病灶中 HIF-1a（缺氧诱导因子 1 α）表达的上调以及随后激活促炎性 M1 巨噬细胞和抗炎性 M2 巨噬细胞。这些发现表明 HIF-1a 激活是改善根尖周伤口愈合的关键因素。该提议的中心假设是，HIF-1a 激活通过诱导 M1 巨噬细胞激活，随后出现 M2 促进病变的最终清除，从而导致根尖周伤口愈合。相反，HIF-1a 激活失败会导致 M1 激活失败，随后 M2 巨噬细胞激活延迟和极化，从而导致慢性炎症和根性肉芽肿。因此，HIF-1a介导的反应是开发改善根尖周伤口愈合的新疗法的目标途径。该探索性提案的总体目标是确定 HIF-1a 激活是否会在免疫活性 (WT) 小鼠中可预测地诱发根尖周病变。该目标将通过两个具体目标来实现。目的 1 是评估 HIF-1a 调节对 WT 小鼠根尖周病变形成和愈合的影响。编码 HIF-1a shRNA 和 HIF-1a 组成型活性形式的慢病毒载体将用于调节 HIF-1a 活性。目的 2 是确定内源性 HIF-1a 对体内巨噬细胞活化和表型的作用。 HIF-1a 在 dKO 小鼠中将失活，并且内源性 HIF-1a 将通过化学抑制脯氨酰羟化酶来激活，脯氨酰羟化酶对于 HIF-1 激活至关重要，以便测量对巨噬细胞激活和感染过程的影响。拟议的研究将提供有关 HIF-1a 在根尖周病变中基本作用的新信息，这是一种涉及 M1/M2 顺序巨噬细胞激活的新型伤口愈合途径，并证明 HIF-1a 在牙槽炎中的治疗潜力。这些结果旨在制定新的治疗策略，以改善牙槽感染的愈合过程，这将在其他慢性纤维化和肉芽肿疾病中具有潜在的应用。