Ad5 Fiber Entry and Trafficking in Lacrimal Acini

Ad5 纤维在泪腺泡中的进入和运输

• 批准号: 7447508
• 负责人: Sarah F Hamm-Alvarez
• 金额: $ 3.65万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-03 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7447508
• 关键词: Acinar Cell; Acinus organ component; Adenoviruses; Affect; American; Antisense Oligonucleotides; Autoimmune Diseases; Autoimmune Process; Binding; Capsid; Capsid Proteins; Cells; Coxsackie Viruses; Cytosol; DNA; Data; Dependence; Development; Diagnosis; Disease; Disease Progression; Distant; Drug Delivery Systems; Endocytosis; Etiology; Eye diseases; Fiber; Functional disorder; Future; Goals; Heparin; Heparitin Sulfate; Histocompatibility Antigens Class I; Intracellular Membranes; Knowledge; Lacrimal gland structure; Lead; Liquid substance; Mediating; Membrane; Molecular; Morbidity - disease rate; Oligonucleotides; Oryctolagus cuniculus; Participant; Pathway interactions; Peptide antibodies; Peptides; Pharmaceutical Preparations; Proteins; Recombinants; Relative (related person); Research; Serotyping; Site; Sjogren' s Syndrome; Small Interfering RNA; Surface; Symptoms; System; Testing; Therapeutic; Viral; Virus; Woman; Work; adenovirus penton protein; adenovirus receptor; base; eye dryness; glycosaminoglycan receptor; insight; lacrimal; lacrimal acini; next generation; novel; ocular surface; polysulfated glycosaminoglycan; prospective; receptor; receptor binding; receptor function; remediation; research study; therapeutic target; trafficking; uptake

A major contribution to ocular morbidity is lacrimal dysfunction, affecting over 10 million Americans. The principal cell of the lacrimal gland and primary contributor of proteins into ocular surface fluid is the lacrimal acinar cell, which is the target of much of the ocular research into the etiology of dry eye diseases including the severe autoimmune disease, Sjogren's syndrome. Ongoing studies are now shedding insights into the precise mechanisms involved in initiation, development and progression of disease, suggesting that dentification of prospective therapeutic targets is likely in the not so distant future. However, we are still very limited in our ability to specifically target the next generation of macromolecular drugs, particularly DNA-, protein- or peptide based drugs, to the lacrimal gland, raising the possibility that we may soon identify advanced therapies for treatment of severe dry eye diseases but will be unable to deliver these drugs to the target site. Our focus here is to explore the unusual and possibly unique uptake mechanism utilized for lacrimal acinar internalization of adenovirus serotype 5 (Ad5). We have demonstrated in lacrimal acinar cells that Ad5 utilizes a unique fiber-dependent internalization pathway, in contrast to the penton-dependent internalization described in other systems. We hypothesize that Ad5 may use multiple fiber receptors for binding and entry in lacrimal acini, and further that one or more of these entry pathways is either unusually robust or unique to lacrimal acini, explaining the unusual fiber-dependence of entry in tandem with the high efficiency of viral transduction in lacrimal acini. We propose to characterize the participants in this novel fiber-dependent internalization pathway at the molecular level, with a particular focus on coxsackievirus adenovirus receptor, major histocompatibility complex class 1 and heparin sulfate-glycosaminoglycan receptors, and their different modes of endocytosis. The aims are as follows: Aim #1. Does coxsackievirus adenovirus receptor mediate fiber-dependent Ad5 internalization or free fiber internalization in lacrimal acini? Aim #2. What other receptors participate in fiber-dependent Ad5 internalization or free fiber internalization in lacrimal acini? Aim #3. What intracellular trafficking pathways mediate fiber-dependent Ad5 internalization or free fiber internalization in lacrimal acini? At the conclusion of this work, we will have elucidated the contributions of specific receptors and endocytic internalization pathways responsible for fiber-dependent internalization of Ad5. We will have determined to what extent the recombinant fiber or knob, the terminal region of the fiber protein, can recapitulate these pathways. Finally, we will have tested proof-of-principle experiments to determine whether fiber or knob can facilitate entry of antisense oligonucleotides and proteins into lacrimal acinar cytosol.

对眼部发病率的主要贡献是泪功能障碍，影响了超过1000万美国人。这 泪腺的主要细胞和蛋白质对眼表液的主要因素是泪 腺泡细胞，这是对干眼症病因的大部分眼部研究的靶标 严重的自身免疫性疾病，Sjogren综合征。现在正在进行的研究正在洞悉 疾病的启动，发展和发展涉及的精确机制，表明 预期治疗靶标的牙齿化可能在不久的将来。但是，我们仍然非常 我们专门针对下一代大分子药物的能力，尤其是DNA-， 基于蛋白质或肽的药物，到泪腺，提高了我们很快识别的可能性 用于治疗严重干眼疾病的晚期疗法，但将无法将这些药物输送到 目标站点。我们在这里的重点是探索用于用于 腺病毒血清型5（AD5）的泪腺泡内在化。我们已经在泪腺细胞中证明了 AD5使用了独特的纤维依赖性内在化途径，与Penton依赖性相反 其他系统中描述的内在化。我们假设AD5可能会使用多个纤维受体进行 在泪acini中结合和进入，进一步，这些进入途径中的一个或多个是异常的 坚固或独特的泪acini，解释了与高的纤维依赖性相关的纤维依赖性 泪液中病毒转导的效率。我们建议在这本小说中表征参与者 纤维依赖性的内在化途径在分子水平上，特别关注Coxsackievivirus 腺病毒受体，主要的组织相容性复合物1类和硫酸硫酸盐 - 糖胺聚糖 受体及其不同的内吞作用模式。目的如下： 目标＃1。 Coxsackievivirus腺病毒受体是否介导了纤维依赖的AD5内在化或游离纤维 泪acini的内在化？ 目标＃2。其他受体参与纤维依赖性AD5内在化或自由纤维内在化中的哪些受体 泪acini？ 目标＃3。哪种细胞内运输途径介导纤维依赖性AD5内在化或游离纤维 泪acini的内在化？ 在这项工作结束时，我们将阐明特定受体和内吞作用的贡献 内部化途径负责AD5的纤维依赖性内在化。我们将决心 重组纤维或旋钮是纤维蛋白的末端区域的多大程度，可以概括这些 途径。最后，我们将进行测试的原理证明实验，以确定纤维还是旋钮是否可以 有助于将反义寡核苷酸和蛋白质进入泪液中的细胞质。