Clinical Investigations In Infectious And Autoimmune Dis

感染性和自身免疫性疾病的临床研究

• 批准号: 6966494
• 负责人: SHARON M WAHL
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6966494
• 关键词: Sjogren' s syndrome; aphthous stomatitis; autoimmune disorder; autoimmunity; biopsy; complementary DNA; human subject; immunodeficiency; inflammation; oral mucosa; periodontium disorder; polymerase chain reaction; wound healing

The objectives of this program focus on the cellular, molecular and biochemical characterization of the underlying pathologic events in infectious and autoimmune diseases, with the long-term intent to develop novel approaches to therapeutic intervention. Nonhealing mucosal and cutaneous lesions due to infection, inflammation or neoplasia can become chronic and debilitating. Despite the profound health effects that chronic wound healing problems and excessive scarring pose, the mechanisms underlying normal and pathologic healing are little understood. Moreover, mucosal epithelial healing is typically more rapid and nearly scarless, prompting a comparison of cutaneous and oral mucosal wound repair to identify controlling factors which may be of benefit in clinical delayed healing responses. In recent studies, not only were the kinetics, cellular composition, and outcome of the healing response found to be substantially altered between cutaneous and oral wounds, but secretory leukocyte protease inhibitor (SLPI) was identified as a pivotal regulatory factor in oral wounds. Age, hormones and immune deficiency play a significant role in inflammatory lesion repair, both in terms of rate of healing and the degree of scarring incurred, attributed in part, to altered inflammatory responses in the aged, with a predominance of neutrophils, excess elastase and a delay in monocyte influx. The delayed cutaneous healing response can be mimicked in animal models including ovariectomized rodents and SLPI knockout mice. Using these pre-clinical models, delayed healing can be reversed by the topical application of TNF antagonists (TNFalpha antibody, thalidomide), recombinant SLPI and synthetic fibronectin peptides, which block downstream inflammatory events and disrupt signal transduction pathways to ameliorate the overzealous leukocyte recruitment and promote matrix degradation. Based on the successful topical delivery of these and other agents in reversing nonhealing wounds, additional novel approaches for inhibiting the persistent leukocyte recruitment and overabundant elastase released at the inflammatory site are being evaluated for their efficacy in facilitating the healing process. Tissue injury in the oral mucosa of humans also activates a cascade of transcriptional events important during the healing process that are not yet clearly defined. To characterize these events and identify potential gene targets for future studies, we used cDNA expression arrays in a clinical model of tissue injury. Mucosal biopsies were taken before 3rd molar extraction, 2-4 hours post-operatively, or at 48 hours. Hybridization patterns were analyzed and validated using real-time PCR. Prior to extraction, the biopsied mucosal tissues were characterized by a panoply of genes that were constitutively expressed. After injury, analysis revealed differential expression of genes involved in transcription, inflammation and remodeling. At 2-4 hours after injury, genes such as Fos, Jun and early growth response protein were upregulated, while genes responsible for inter-cellular adhesion were downregulated. At 48 hours after injury, the gene profile had shifted towards tissue remodeling. In these new studies, we identify genes constitutively expressed in normal oral mucosa and transcriptional events following mucosal tissue injury, which may be useful in identifying new therapeutic targets. In clinically-derived chronic inflammatory lesions, perpetuated by bacteria and/or their products (periodontal tissues, etc.), viruses (HIV-1), other pathogens, or autoimmune responses (Sjogren's Syndrome), characterization of the systemic immune response, as well as the lesion cellular constituents using laser capture microdissection, and proteome and transcriptome responses (cDNA expression array), offers new insight into targets for amelioration of tissue degradation and delayed repair.

该计划的目标集中在感染性和自身免疫性疾病中潜在病理事件的细胞，分子和生物化学表征，长期意图是开发新的治疗干预方法。由于感染、炎症或瘤形成而导致的不愈合粘膜和皮肤损伤可变为慢性和使人衰弱的。尽管慢性伤口愈合问题和过度瘢痕形成对健康有深远的影响，但对正常和病理愈合的机制知之甚少。此外，粘膜上皮愈合通常更快且几乎无瘢痕，这促使比较皮肤和口腔粘膜伤口修复，以确定可能有益于临床延迟愈合反应的控制因素。在最近的研究中，不仅是动力学，细胞组成，和愈合反应的结果被发现在皮肤和口腔伤口之间的实质性改变，但分泌性白细胞蛋白酶抑制剂（SLPI）被确定为一个关键的调节因子在口腔伤口。年龄、激素和免疫缺陷在炎症性病变修复中起重要作用，无论是在愈合率和瘢痕形成程度方面，部分归因于老年人炎症反应的改变，中性粒细胞占主导地位，弹性蛋白酶过量和单核细胞流入延迟。延迟的皮肤愈合反应可以在包括卵巢切除的啮齿动物和SLPI敲除小鼠的动物模型中模拟。使用这些临床前模型，通过局部应用TNF拮抗剂（TNF α抗体、沙利度胺）、重组SLPI和合成纤连蛋白肽可以逆转延迟愈合，这些拮抗剂阻断下游炎症事件并破坏信号转导途径以改善过度活跃的白细胞募集并促进基质降解。基于这些和其他药物在逆转不愈合伤口中的成功局部递送，正在评估用于抑制持续性白细胞募集和在炎症部位释放的过量弹性蛋白酶的其他新方法在促进愈合过程中的功效。 人类口腔粘膜中的组织损伤也激活了在愈合过程中重要的级联转录事件，这些事件尚未明确定义。为了表征这些事件并确定未来研究的潜在基因靶点，我们在组织损伤的临床模型中使用cDNA表达阵列。在第三磨牙拔除前、术后2-4小时或48小时进行粘液组织活检。使用实时PCR分析和验证杂交模式。在提取之前，活检的粘膜组织的特征在于组成型表达的一系列基因。损伤后，分析显示参与转录、炎症和重塑的基因的差异表达。损伤后2-4 h，Fos、Jun和早期生长反应蛋白等基因表达上调，而细胞间粘附相关基因表达下调。在损伤后48小时，基因谱已经转向组织重塑。在这些新的研究中，我们确定了在正常口腔粘膜中组成型表达的基因和粘膜组织损伤后的转录事件，这可能有助于确定新的治疗靶点。 在由细菌和/或其产物（牙周组织等）持续存在的临床来源的慢性炎性病变中，病毒（HIV-1）、其他病原体或自身免疫反应（Sjogren综合征），使用激光捕获显微切割以及蛋白质组和转录组反应（cDNA表达阵列）表征全身免疫反应以及病变细胞成分，为改善组织降解和延迟修复的靶点提供了新的见解。