Modeling Novel Treatments for Dry Eye

干眼症新疗法建模

• 批准号: 7250497
• 负责人: Gordon William Laurie
• 金额: $ 56.58万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7250497
• 关键词: Acinar Cell; Acute; Address; Affect; Alternative Splicing; Animal Model; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Autoantibodies; Autoimmune Process; Binding; Binding Sites; Biological; Biological Assay; Biology; Biomanufacturing; Brain; CA-125 Antigen; CASP8 and FADD-like apoptosis regulating protein; Carbohydrates; Caspase; Cell Death; Cell Survival; Cell surface; Cells; Cessation of life; Chronic; Collaborations; Complex; Cornea; Cultured Cells; Cyclosporine; Cyclosporins; Data; Detection; Development; Disease; Drug Prescriptions; Dry Eye Syndromes; Educational process of instructing; Engineering; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Epithelium; Epitopes; Exons; Experimental Animal Model; Eye; Eye diseases; Faculty; Funding; G Protein-Coupled Receptor Genes; Gelatinase B; Goals; Human; Individual; Inflammation; Inflammatory; Interleukin-1; Lacrimal gland structure; Link; Liquid substance; MAPK8 gene; Mediator of activation protein; Methods; Mitogens; Modeling; Molecular; Mucins; Mus; Muscarinic M3 Receptor; N-terminal; NK Cell Activation; Oryctolagus cuniculus; Pathway interactions; Patients; Pharmacologic Substance; Phase; Phosphotransferases; Population; Preclinical Testing; Principal Investigator; Protein Engineering; Proteins; RNA Splicing; Recombinant Proteins; Research; Research Personnel; Saliva; Salivary Glands; Serum; Signal Pathway; Signal Transduction; Site; Sjogren' s Syndrome; Small Business Technology Transfer Research; Small Interfering RNA; Students; Testing; Tetanus Helper Peptide; Thinking; Time; Topical application; Toxic effect; Transgenes; Translations; Universities; Variant; Virginia; Work; autocrine; base; caspase-8; conjunctiva; corneal epithelium; cytokine; day; deletion analysis; enhancing factor; eye dryness; heparanase; human FRAP1 protein; inhibitor/antagonist; lacrimal; medical schools; meibomian gland; mutant; novel; novel therapeutics; ocular surface; ovarian neoplasm; preclinical study; prescription document; prescription procedure; prevent; programs; receptor; research study; syndecan; tear proteins; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): Our multi-disciplinary and multi-institutional application addresses dry eye syndromes from the perspective of the new human prosecretory mitogen 'lacritin' discovered by us. Lacritin is restrictively expressed in lacrimal and meibomian glands and in the cornea and conjunctiva, where it appears to be capable of protecting epithelia of the lacrimal-corneal axis against inflammation-associated cell death. Increased levels of proinflammatory cytokine TNFa in tears of dry eye patients is associated with damage to ocular surface epithelia. Indeed, adding TNFa to cultured human corneal epithelial cells promotes death via caspases-8 and -3. Recently we observed that caspase activation and death is completely prevented by inclusion of 10 nM lacritin. This observation has been reinforced by lacritin deletion analysis that reveals a cytoprotective site within lacritin's C-terminus. Lacritin's utilizes a unique cell targeting mechanism: heparanase unblocks a lacritin binding site within the N-terminal ectodomain of cell surface syndecan-1. Bound lacritin is then likely presented to a GPCR. Since heparanase has a lower pH optimum, it is possible that lacritin is protective against the hypothetical sudden low pH 'danger signal' thought to underlie the initiation of primary Sjogren's syndrome dry eye in some individuals. In rabbit preclinical studies, topical application of lacritin promotes increased tear flow for at least 4 hr without toxicity - even over 30 days of continuous treatment. In cell culture, lacritin stimulates tear secretion by lacrimal acinar cells - the same cells from which it is secreted. It also promotes corneal epithelial MUC16 and lacritin expression. Since lacritin is a natural tear protein, this suggests mechanisms of upstream and downstream autocrine stimulation that prolong lacritin's cytoprotective and prosecretory effects. Tear proteins likely function as bioactive complexes. These activities can be harnessed by recombinant protein engineering. Our working hypothesis is that lacritin is naturally protective against dry eye inflammation. Our immediate goal is to optimize lacritin's cytoprotective activity and understand its mechanism of action. Our first aim is to engineer the smallest and most cytoprotective form of lacritin. Our second aim is to work out biological pathways that underlie its cytoprotective activity in a search for treatment synergies or counterindications. Our third aim is to preclinically test topically applied or genetically induced lacritin in animal models of dry eye.

描述（由申请人提供）：我们的多学科和多机构申请从我们发现的新人类分泌促细胞分裂原“乳泌素”的角度解决干眼综合症。泪素在泪腺和睑板腺以及角膜和结膜中受到限制性表达，它似乎能够保护泪腺-角膜轴的上皮细胞免受炎症相关的细胞死亡。干眼症患者泪液中促炎细胞因子 TNFa 水平升高与眼表上皮损伤有关。事实上，向培养的人角膜上皮细胞中添加 TNFa 可通过 caspase-8 和 -3 促进细胞死亡。最近我们观察到，加入 10 nM 乳泌素可以完全阻止 caspase 激活和死亡。乳泌素缺失分析进一步证实了这一观察结果，该分析揭示了乳泌素 C 末端内的细胞保护位点。 Lacritin 利用独特的细胞靶向机制：乙酰肝素酶可解锁细胞表面 syndecan-1 N 末端胞外域内的 lacritin 结合位点。然后结合的泪泌素可能被呈递给 GPCR。由于乙酰肝素酶的最佳 pH 值较低，因此乳泌素可能可以防止假设的突然低 pH“危险信号”，该信号被认为是某些个体引发原发性干燥综合征干眼的基础。在兔子临床前研究中，局部应用泪泌素可促进泪液流量增加至少 4 小时且无毒性 - 甚至连续治疗超过 30 天。在细胞培养中，泪泌素刺激泪腺腺泡细胞分泌泪液，泪腺腺细胞与分泌泪泌素的细胞相同。它还促进角膜上皮 MUC16 和泪泌素的表达。由于泪泌素是一种天然的泪液蛋白，这表明上游和下游自分泌刺激的机制可以延长泪泌素的细胞保护和促分泌作用。泪液蛋白可能作为生物活性复合物发挥作用。这些活性可以通过重组蛋白质工程来利用。我们的工作假设是，泪泌素具有天然的预防干眼炎症的作用。我们的近期目标是优化乳泌素的细胞保护活性并了解其作用机制。我们的首要目标是设计出最小且最具细胞保护作用的泪泌素形式。我们的第二个目标是找出其细胞保护活性的生物学途径，以寻找治疗协同作用或反适应症。我们的第三个目标是在干眼动物模型中对局部应用或基因诱导的泪泌素进行临床前测试。