Normal And Pathologic Mechanisms Of Inflammation, Innate And Acquired Immunity

炎症、先天性和获得性免疫的正常和病理机制

• 批准号: 7967008
• 负责人: SHARON M WAHL
• 金额: $ 163.29万
• 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/7967008
• 关键词: ANXA2 gene; Affect; Alteplase; Animal Model; Anthracenes; Anti-Inflammatory Agents; Anti-inflammatory; Antigen-Presenting Cells; Asthma; Autoantibodies; Autoimmune Diseases; Autoimmune Process; Autoimmune Responses; Autoimmunity; Automobile Driving; Bacteria; Basic Science; Binding Proteins; Biochemical; Biological Markers; Biopsy; Blood Circulation; Bone Resorption; C57BL/6 Mouse; Cardiovascular Diseases; Cell Lineage; Cells; Chronic; Clinical; Coagulation Process; Collaborations; Complement Activation; Complex; Coronary Arteriosclerosis; DNA Microarray Chip; Data; Dendritic Cells; Dentition; Development; Disease; Docking; Etanercept; Etiology; Event; Exocrine Glands; Experimental Animal Model; Failure; Fibrinolysis; Gene Expression; Gene Expression Profile; Generations; Genes; Head and Neck Squamous Cell Carcinoma; Healed; Host Defense; Immune; Immune response; Immunity; Immunosuppressive Agents; Impaired wound healing; Incidence; Infection; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injury; Interleukin-17; Interleukin-6; Kinetics; Knockout Mice; Lesion; Life; Link; Low Birth Weight Infant; Lung; Lymphoid Cell; Malignant Neoplasms; Modeling; Molecular; Mouth Diseases; Mus; Myelogenous; Myeloid Cells; Nature; Neoplasm Metastasis; Oral mucous membrane structure; Organism; Papilloma; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Periodontal Diseases; Peripheral; Pharmaceutical Preparations; Physiological; Plasmin; Plasminogen; Population; Porphyromonas gingivalis; Predisposition; Premature Birth; Process; Production; Protective Agents; Proteomics; Regulation; Regulation of Proteolysis; Relative (related person); Research Personnel; Role; Salivary Glands; Seminal; Signal Pathway; Sjogren' s Syndrome; Skin Papilloma; Squamous cell carcinoma; Syndrome; System; T-Lymphocyte; T-Lymphocyte Subsets; Therapeutic Intervention; Tissues; Transforming Growth Factor beta; Tumor Cell Invasion; Tumor Immunity; Virus; Wound Healing; acquired immunity; antileukoprotease; base; chemokine; cytokine; healing; human SLPI protein; immune function; immunopathology; inhibitor/antagonist; insight; interleukin-23; laser capture microdissection; macrophage; malignant mouth neoplasm; microorganism; migration; monocyte; neoplastic; novel strategies; oral pathogen; osteoclastogenesis; outcome forecast; prevent; programs; receptor-mediated signaling; repaired; response; skin squamous cell carcinoma; tumor; tumorigenesis

SLPI regulation of proteolytic cascade in inflammation and cancer (40%) Enhanced inflammation, delayed healing and susceptibility to infection in the absence of SLPI suggested a link between SLPI and regulation of inflammatory responses. Following identification of annexin A2 as a SLPI-binding protein and that annexin A2 serves as a docking station for tissue-type plasminogen activator (tPA) and plasminogen to facilitate plasminogen activation and generation of plasmin, we demonstrated that the SLPI-annexin A2 interaction interrupts the proteolytic cascade responsible for plasmin generation. Moreover, we have shown that in SLPI null mice, macrophages process unregulated levels of plasmin, which contributes to excess tissue degradation, increased inflammation and failed healing following tissue injury. By inhibiting plasmin, which has a myriad of downstream consequences relative to complement activation, fibrinolysis, cytokine regulation, TGF-β activation and other inflammatory sequelae, SLPI can exert control over proteolytic and inflammatory activation networks. Because fibrinolytic processes are fundamental not only to inflammation, coagulation and tissue repair, but growing evidence highlights their dysregulation in the pathogenesis of multiple chronic inflammatory disorders, cardiovascular disease and malignancy, further defining and regulating these pathways is of major significance. Relevant to these findings, we have explored the role of SLPI as a protective agent in metastatic head and neck squamous cell carcinoma (HNSCC), which remains one of the 10 most frequently occurring cancers worldwide with a very poor prognosis, in collaboration with investigators at UMD. Our data imply that SLPI has anti-tumor activities, which may include interference with requisite proteolytic steps underlying tumor cell invasion and affecting the cell-associated plasminogen activation system, suggesting a strategy to predict (biomarker) and prevent (therapy) tumor invasive potential in oral cancer. In a model of squamous cell carcinoma in SLPI null and WT C57BL/6 mice, our evidence confirms that the absence of SLPI is associated with more rapid and extensive development of tumors, enabling characterization of the cellular and molecular pathways involved. Utilizing the 7,12-dimethylbenzαanthracene (DMBA)-induced murine skin squamous cell carcinoma (SCC) model in exploring SLPIs function in tumor development and progression, evidence supported a role for SLPI in the kinetics and incidence of skin papilloma, and in significantly inhibiting lung metastasis. However, SLPI did not appear to influence the conversion of papilloma to cancer, thereby focusing our efforts on its role in invasion, migration and metastasis. Autoimmunity and Sjogrens syndrome(SS) (30%) An important objective of our studies is to define the underlying mechanisms by which inflammatory and immune responses go awry, resulting in autoimmune pathogenesis, as exemplified by Sjogren's syndrome. The autoimmune hallmarks of SS include focal lymphoid cell infiltration of the exocrine glands (focus score) and the production of autoantibodies. The etiology of SS remains largely unknown and a conundrum is presented by the failure of many drugs that are effective in other autoimmune disorders, particularly TNFα antagonists, such as etanercept. A potential clue to the lack of efficacy of TNFα inhibitors is the elevated IL-17 in SS patients peripheral circulation, which is also unmodulated by anti-TNFα therapy. In salivary gland biopsies, we performed gene profiling to correlate specific gene expression with T cell subsets and/or their products within the inflamed tissues. At the cellular level, we have shown that expression of IL-17 and also TGF-β progressively increases with higher biopsy focus scores. Thus, we have connected IL-17 to SS tissue pathology, along with several cytokines that have been shown to be supporting molecules for development of this lineage (TGF-β, IL-23 and IL-6) without a corresponding increase in Treg with severe disease. This imbalance between pro- and anti-inflammatory forces may perpetuate the autoimmune lesions. Infectious inflammation and injury in oral mucosa (30%) Chronic periodontal diseases, which are complex, inflammatory oral diseases involving gram-negative microorganisms such as P. gingivalis (Pg), are characterized by destruction of supporting tissues surrounding the dentition. However, it has become increasingly clear that while the presence of these organisms is central to pathogenesis, the destructive process is largely host-derived. Given the seminal role of antigen presenting cells (APC) in innate and adaptive immunity, and the common lineage of monocytes, macrophages and dendritic cells, we exposed these cells to Pg and identified a Pg LPS-inducible convergent, transcriptional core response among these populations, reflected by a shared, but quantitatively distinct proteomic response. Nonetheless, despite these similarities, clear differences emerged between the myeloid populations. The finding that long-lived myeloid inflammatory cells, particularly DC, rapidly and aggressively respond to Pg LPS with generation of chemokines, proteases and cytokines capable of driving T helper cell lineage (Th17) polarization without evidence of corresponding immunosuppressive pathways highlights their prominent role in host defense and progressive tissue pathogenesis. The shared, unique and/or complementary transcriptional and proteomic profiles may frame the context of the host response to P. gingivalis, contributing to the destructive nature of periodontal inflammation, including osteoclastogenesis. In pursuing this link, we are further examining the interaction between intact Pg bacteria (both fimbriated and non-fimbriated) and myeloid cells to delineate receptor-mediated signaling pathways linked to osteoclastogenesis and bone resorption. These studies are of additional importance since P. gingivalis, once considered to be primarily an oral pathogen, has more recently been linked to several systemic conditions, such as coronary artery disease and preterm delivery of low birth weight infants.

SLPI 对炎症和癌症中蛋白水解级联的调节 (40%) 在没有 SLPI 的情况下，炎症增强、愈合延迟和对感染的易感性表明 SLPI 与炎症反应调节之间存在联系。在鉴定膜联蛋白 A2 为 SLPI 结合蛋白并且膜联蛋白 A2 作为组织型纤溶酶原激活剂 (tPA) 和纤溶酶原的对接站以促进纤溶酶原激活和纤溶酶生成后，我们证明 SLPI-膜联蛋白 A2 相互作用会中断负责纤溶酶生成的蛋白水解级联。此外，我们还发现，在 SLPI 缺失小鼠中，巨噬细胞处理不受调节水平的纤溶酶，这会导致组织过度降解、炎症增加以及组织损伤后无法愈合。通过抑制纤溶酶（纤溶酶具有与补体激活、纤维蛋白溶解、细胞因子调节、TGF-β激活和其他炎症后遗症相关的众多下游后果），SLPI 可以控制蛋白水解和炎症激活网络。由于纤溶过程不仅是炎症、凝血和组织修复的基础，而且越来越多的证据强调纤溶过程在多种慢性炎症性疾病、心血管疾病和恶性肿瘤的发病机制中的失调，因此进一步定义和调节这些途径具有重要意义。与这些发现相关，我们与 UMD 的研究人员合作，探讨了 SLPI 作为转移性头颈鳞状细胞癌 (HNSCC) 保护剂的作用，这种癌症仍然是全球 10 种最常见的癌症之一，预后非常差。我们的数据表明 SLPI 具有抗肿瘤活性，其中可能包括干扰肿瘤细胞侵袭所需的蛋白水解步骤并影响细胞相关的纤溶酶原激活系统，这提出了一种预测（生物标志物）和预防（治疗）口腔癌肿瘤侵袭潜力的策略。在 SLPI 缺失和 WT C57BL/6 小鼠的鳞状细胞癌模型中，我们的证据证实，SLPI 的缺失与肿瘤更快、更广泛的发展相关，从而能够表征所涉及的细胞和分子途径。利用 7,12-二甲基苯并蒽 (DMBA) 诱导的小鼠皮肤鳞状细胞癌 (SCC) 模型探索 SLPI 在肿瘤发生和进展中的功能，证据支持 SLPI 在皮肤乳头状瘤的动力学和发病率以及显着抑制肺转移中的作用。然而，SLPI 似乎并不影响乳头状瘤向癌症的转化，因此我们将重点放在其在侵袭、迁移和转移中的作用。 自身免疫和干燥综合征 (SS) (30%) 我们研究的一个重要目标是确定炎症和免疫反应出错并导致自身免疫发病机制的潜在机制，如干燥综合征所示。 SS 的自身免疫标志包括外分泌腺的局灶性淋巴细胞浸润（病灶评分）和自身抗体的产生。 SS 的病因在很大程度上仍不清楚，并且许多对其他自身免疫性疾病有效的药物（特别是 TNFα 拮抗剂，如依那西普）均失败，这是一个难题。 TNFα 抑制剂缺乏疗效的一个潜在线索是 SS 患者外周循环中 IL-17 升高，而抗 TNFα 治疗也无法调节该水平。在唾液腺活检中，我们进行了基因分析，将特定基因表达与发炎组织内的 T 细胞亚群和/或其产物相关联。在细胞水平上，我们发现 IL-17 和 TGF-β 的表达随着活检焦点分数的升高而逐渐增加。因此，我们将 IL-17 与 SS 组织病理学以及已被证明是该谱系发育的支持分子的几种细胞因子（TGF-β、IL-23 和 IL-6）联系起来，而严重疾病时 Treg 细胞不会相应增加。促炎和抗炎力量之间的这种不平衡可能会使自身免疫病变永久存在。 口腔粘膜感染性炎症和损伤（30%） 慢性牙周病是一种复杂的炎症性口腔疾病，涉及革兰氏阴性微生物，例如牙龈卟啉单胞菌（Pg），其特征是牙列周围支持组织的破坏。然而，越来越清楚的是，虽然这些生物体的存在是发病机制的核心，但破坏过程很大程度上是源自宿主的。鉴于抗原呈递细胞 (APC) 在先天性和适应性免疫中的重要作用，以及单核细胞、巨噬细胞和树突状细胞的共同谱系，我们将这些细胞暴露于 Pg 中，并在这些群体中鉴定出 Pg LPS 诱导的聚合转录核心反应，这反映在共享但数量上不同的蛋白质组反应上。尽管如此，尽管有这些相似之处，骨髓细胞群之间还是出现了明显的差异。研究发现，长寿命的骨髓炎症细胞，特别是 DC，能够快速、积极地对 Pg LPS 作出反应，产生能够驱动 T 辅助细胞谱系 (Th17) 极化的趋化因子、蛋白酶和细胞因子，而没有相应的免疫抑制途径的证据，这突显了它们在宿主防御和进行性组织发病机制中的突出作用。共享的、独特的和/或互补的转录和蛋白质组谱可能构成了宿主对牙龈卟啉单胞菌反应的背景，有助于牙周炎症的破坏性，包括破骨细胞生成。在研究这种联系的过程中，我们正在进一步研究完整的 Pg 细菌（包括菌毛和非菌毛）和骨髓细胞之间的相互作用，以描绘与破骨细胞生成和骨吸收相关的受体介导的信号通路。这些研究具有额外的重要性，因为牙龈卟啉单胞菌曾经被认为主要是一种口腔病原体，最近被认为与多种全身性疾病有关，例如冠状动脉疾病和低出生体重婴儿的早产。