Unfolded Protein Response as a Therapeutic Target for ADRP Animal Models

未折叠蛋白反应作为 ADRP 动物模型的治疗靶点

• 批准号: 8288847
• 负责人: Marina Gorbatyuk
• 金额: $ 3.53万
• 依托单位: UNIVERSITY OF NORTH TEXAS HLTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8288847
• 关键词: 11 cis Retinal; Accounting; Affect; Animal Model; Antibodies; Apoptosis; Apoptotic; Biological Preservation; Blindness; Caspase; Cell Death; Cessation of life; Comparative Study; Contrast Sensitivity; Data; Deterioration; Development; Disease; Electroretinography; Endoplasmic Reticulum; Ethnic group; Evaluation; GADD45; Gene Transfer; Genes; Genetic Variation; Goals; Golgi Apparatus; Inherited; Investigational Drugs; Knockout Mice; Lead; Location; Measures; Mediating; Messenger RNA; Methods; Modeling; Molecular; Molecular Chaperones; Monitor; Mus; Mutation; Nucleosomes; Opsin; Optometry; Pathogenesis; Pathway interactions; Patients; Photoreceptors; Proteins; Rattus; Resistance; Retina; Retinal; Retinal Degeneration; Retinitis Pigmentosa; Reverse Transcriptase Polymerase Chain Reaction; Rhodopsin; Role; Serotyping; Severities; Signal Transduction; Small Interfering RNA; Small RNA; Structure; Supplementation; Techniques; Testing; Therapeutic; Transgenic Mice; Viral; Virus; Vision; Visual impairment; Vitamin A; X inherited trait; base; biological adaptation to stress; caspase 12; caspase-7; defined contribution; design; dosage; drug discovery; endoplasmic reticulum stress; gene therapy; improved functioning; molecular chaperone GRP78; morphometry; mouse model; mutant; novel; programs; promoter; protein misfolding; protein transport; response; success; therapeutic target; trafficking; transcription factor CHOP; vector

Unfolded Protein Response as a Therapeutic Target for ADRP Animal Models This project is focused on the elucidation of the role of the Unfolded Protein Response (UPR) in autosomal dominant retinitis pigmentosa (ADRP) pathogenesis and development of the gene therapy based on modulation of the UPR signaling markers. Retinitis pigmentosa (RP) is the most common inherited form of blindness, affecting about 1 in every 4000 people in all ethnic groups worldwide. RP can be transmitted either as an autosomal dominant (ADRP), autosomal recessive (ARRP), or X-linked trait. More than 100 mutations in rhodopsin account for approximately 30% of ADRP cases with varying severity of visual impairment. Misfolded opsin interferes with the trafficking of wild-type rhodopsin, accumulates in the endoplasmic reticulum (ER) and stimulates a signal transduction cascade known as the Unfolded Protein Response (UPR). If unchecked, this pathway triggers photoreceptor death, presumably through apoptosis. Although supplementation with vitamin A may be beneficial in some cases, currently, there is no effective pharmacological therapy for ADRP. Therefore, the major objective of this proposal is to determine whether the gene therapy based on the re-programming of the ER stress response caused by aberrant rhodopsin is a viable treatment, unlimited by different localizations of rhodopsin mutations (P23H and T17M). In two mouse models of ADRP, we plan to reprogram the ER stress signaling by viral delivery of the molecular chaperone GRP78/BiP and delivery of small interfering siRNAs targeting caspase-7, caspase-12 and pro-apoptotic CHOP/GADD153 mRNAs to diminish the level of apoptosis in ADRP photoreceptors. For each ADRP model, we plan to: (1) modulate the UPR in favor of activation of pro-survival pathway by over- expression of BiP protein; (2) suppress apoptosis by diminishing levels of activated caspase-7 and caspase-12 and (3) inhibit the CHOP-associated apoptosis by targeting CHOP mRNA. We will monitor survival of photoreceptors using electroretinography and morphometry and will measure the activation of the ER stress and apoptosis using specific antibodies and RT-PCR. We will also measure improvement in vision using Optometry, a technique that can evaluate both acuity and contrast sensitivity in mice. We anticipate that the success of this approach will also require the appropriate combination of AAV serotype, vector dosage, photoreceptor specific promoter and optimized expression for the chaperone BiP. While AAV mediated gene transfer is being developed for treatment of RP, the suppression of ER stress and of apoptosis using chaperones is novel. This approach may overcome the genetic diversity of this disease and reveal the pathways of cell death that lead from mutation to retinal degeneration.

未折叠蛋白应答作为ADRP动物模型的治疗靶点 该项目的重点是阐明未折叠蛋白反应（UPR）在常染色体中的作用。 显性视网膜色素变性（ADRP）的发病机制及基于基因治疗的发展 UPR信号标记的调节。视网膜色素变性（RP）是最常见的遗传形式， 失明，影响全世界所有种族群体中每4000人中约有1人。RP可以通过以下方式传输： 常染色体显性（ADRP）、常染色体隐性（ARRP）或X连锁性状。超过100种突变， 视紫红质占ADRP病例的约30%，具有不同程度的视力损害。错误折叠 视蛋白干扰野生型视紫红质的运输，在内质网（ER）中积累， 刺激称为未折叠蛋白质反应（UPR）的信号转导级联。如果不检查， 这可能是通过细胞凋亡引起的。虽然补充维生素A 在某些情况下可能是有益的，目前，没有有效的药物治疗ADRP。因此，我们认为， 这项建议的主要目的是确定是否基因治疗的基础上重新编程的 由异常视紫红质引起ER应激反应是一种可行的治疗方法，不受不同定位的限制 视紫红质突变（P23 H和T17 M）。 在两种ADRP小鼠模型中，我们计划通过病毒传递ADRP基因来重新编程ER应激信号。 分子伴侣GRP 78/BiP和靶向caspase-7、caspase-12的小干扰siRNA的递送 和促凋亡CHOP/GADD 153 mRNA以降低ADRP光感受器中的凋亡水平。为 每个ADRP模型，我们计划：（1）调节UPR，有利于激活促生存途径，通过过度， BiP蛋白的表达;（2）通过降低活化的caspase-7和caspase-12的水平来抑制凋亡 （3）通过靶向CHOP mRNA抑制CHOP相关凋亡。我们将监测 使用视网膜电描记术和形态测量术测量光感受器，并将测量ER应激的激活 特异性抗体和RT-PCR检测细胞凋亡。我们还将测量视力的改善， 验光，一种可以评估小鼠敏锐度和对比敏感度的技术。我们预计 这种方法的成功还需要AAV血清型，载体剂量， 光感受器特异性启动子和分子伴侣BiP的优化表达。而AAV介导的基因 转移正在被开发用于治疗RP，抑制ER应激和细胞凋亡， chaperones是新奇的。这种方法可以克服这种疾病的遗传多样性，并揭示这种疾病的遗传多样性。 细胞死亡的途径，导致从突变到视网膜变性。