Modeling Novel Treatments for Dry Eye

干眼症新疗法建模

基本信息

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
项目类别：
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&apos 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

项目摘要

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应用程序从我们发现的新的人类起诉有丝分裂原'lacritin'的角度来解决干眼症综合症。酸性在泪中和meibomian腺以及角膜和结膜中限制性地表达，在那里它似乎能够保护富氏轴轴的上皮，以防止与炎症相关的细胞死亡。干眼症患者撕裂中促炎性细胞因子TNFA的水平增加与眼部表面上皮损伤有关。实际上，将TNFA添加到培养的人角膜上皮细胞中促进Caspases -8和-3促进死亡。最近，我们观察到胱天蛋白酶激活和死亡完全通过纳入10 nm的曲息蛋白来预防。通过透眼缺失分析来增强了这一观察结果，该分析揭示了细胞蛋白的C-末端中的细胞保护位点。流质蛋白利用一种独特的细胞靶向机制：肝素酶在细胞表面syndecan-1的N末端外生域内解阻到透明度。然后可能会将结合的透眼素呈现给GPCR。由于肝素酶的最佳pH值较低，因此透眼素有可能对假设突然的低pH“危险信号”的保护作用，这是在某些个体中启动原发性Sjogren综合征干眼的基础的基础。在兔子临床前研究中，局部应用透眼素会促进至少4小时的泪液流动增加，而无需毒性，即使在连续治疗的30天内也会增加。在细胞培养中，透眼素刺激泪腺泡细胞的撕裂分泌 - 与其分泌的相同细胞。它还促进了角膜上皮MUC16和基表达。由于酸性蛋白是一种天然泪液，因此这表明了上游和下游自分泌刺激的机制，可以延长良龙的细胞保护作用和起诉作用。泪蛋白可能充当生物活性复合物。这些活动可以通过重组蛋白工程来利用。我们的工作假设是，酸性蛋白天然可防止眼睛炎症。我们的近期目标是优化流质素的细胞保护活性并了解其作用机理。我们的第一个目的是设计最小，最细胞保护的曲息蛋白。我们的第二个目的是制定生物学途径，这些途径是其在寻找治疗协同作用或反向指标时的细胞保护活性的基础。我们的第三个目的是临时检验在干眼动物模型中局部施用或遗传诱导的卓蛋白。