Defining Barriers to Gene Therapy

定义基因治疗的障碍

• 批准号: 10163849
• 负责人: Stephen H Tsang
• 金额: $ 34.69万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10163849
• 关键词: Ablation; Address; Adult; Affect; Age; Age-Years; Alleles; American; Anabolism; Binding; Catabolism; Cessation of life; Clinical Trials; Combined Modality Therapy; Cone; Cone dystrophy; Cre driver; Data; Degenerative Disorder; Diagnosis; Disease; Electroretinography; Engineering; European; Functional disorder; Funding; GTP-Binding Proteins; Gene therapy trial; Genes; Genetic; Genotype; Glycolysis; Goals; Grant; Guanine Nucleotides; Human; Injections; Intervention; Light; LoxP-flanked allele; Measurement; Mediating; Metabolic; Methods; Modeling; Mus; Mutagenesis; Mutation; Onset of illness; Outcome; Pathologic; Pathway interactions; Patients; Peer Review; Phase; Phenotype; Photoreceptors; Public Health; Publications; Quality of life; Retina; Retinal Cone; Retinal Degeneration; Retinal Diseases; Retinal Dystrophy; Risk; Rod; Sirtuins; Study models; Supplementation; System; Tamoxifen; Testing; Therapeutic; Therapeutic Intervention; Time; Transcription Repressor; Vertebrate Photoreceptors; Viral Vector; Vision; alpha Subunit Transducin; base; brain circuitry; clinically relevant; cost efficient; design; effectiveness evaluation; efficacious treatment; experience; experimental study; gene function; gene therapy; human model; improved; innovation; meetings; metabolome; mouse model; novel; phosphoric diester hydrolase; photoreceptor degeneration; programs; restoration; small molecule; tool

PROJECT SUMMARY Of the retinal degenerative diseases that affect 9 million Americans, cone photoreceptor dystrophies are arguably the most devastating. Gene therapy is a potential means to strengthen photoreceptor viability. However, the first human gene therapy trial for retinal degeneration found improved visual function but did not slow degeneration of photoreceptors. The goal of this gene therapy-oriented proposal is to determine whether therapy is achievable in the context of an already diseased retina and if metabolic reprogramming could be an efficacious treatment option. During the previous funding period, we succeeded in restoring retinal function for more than 11 months in a mouse model of rod degeneration even after the onset of degeneration and at late-stage disease. We now intend to determine whether the same outcomes are achievable in cone-based dystrophies. To do this, we will generate a novel, inducible genetic rescue system in the cone-specific G-protein, guanine nucleotide binding α- transducin 2 (Gnat2), which will allow us to conditionally reverse GNAT2-deficiency while controlling the temporal and spatial aspects of phenotypic reversal. Using Gnat2floxSTOP/Gnat2CreERT2, we will establish that the model faithfully recapitulates cone-mediated dystrophies (Aim 1). We will restore the model to wild type via tamoxifen injection at early, middle, and late disease stages and assess effects on the rate of degeneration (Aim 2) to determine the temporal limitations of gene therapy. Finally, we will induce metabolic reprogramming and assess its utility as a possible non-gene- specific strategy for treating cone degenerations -based dystrophies (Aim 3). The Gnat2floxSTOP/Gnat2CreERT2 programmable model will provide a platform for contributing to ongoing efforts aimed at increasing restoration of visual function following gene therapy for cone-mediated dystrophies. It will also allow us to address several compelling, clinically relevant questions: Is the brain’s circuitry sufficiently plastic to recover from the pathological changes caused by the Gnat2 mutation? Is there a point of no return after which, despite reversion of the genotype to wild type, cones cannot be salvaged? Can temporal barriers to gene therapy be relieved by metabolic reprogramming? Taken together, this proposal is certain to 1) define the factors limiting interventional therapy; 2) validate a new, inducible model of cone-mediated retinal degeneration; and 3) determine whether metabolic reprogramming can serve as an efficacious, non-gene-specific strategy for treating retinal degeneration.

项目摘要 在影响900万美国人的视网膜退行性疾病中 可以说是最毁灭性的。基因治疗是增强感光活性的潜在手段。 然而，第一个永久性变性的人类基因疗法试验发现了视觉功能的改善，但没有 光感受器的缓慢变性。该基因疗法的建议的目的是确定是否是否 在已经剖析的视网膜的背景下，治疗是可以实现的，如果代谢重编程可能是 易于治疗选择。 在上一个资金期间，我们成功地恢复了剩余功能超过11个月 在变性和晚期疾病开始后，在杆变性的小鼠模型中。我们现在 打算确定在基于锥体的营养不良中是否可以实现相同的结果。为此，我们将 在锥体特异性的G蛋白，鸟嘌呤核苷酸结合α-中产生一种新型的，可诱导的遗传救援系统 跨多霉素2（GNAT2），这将使我们能够在控制临时性的同时有条件地逆转GNAT2缺乏效率 和表型逆转的空间方面。 使用gnat2floxstop/gnat2creert2，我们将忠实地概括锥体介导的模型 营养不良（AIM 1）。我们将在早期，中和晚期通过Tamoxifer注入将模型恢复为野生型 疾病阶段和评估对变性率的影响（AIM 2），以确定 基因疗法。最后，我们将诱导代谢重编程，并评估其用作可能的非基因 - 治疗基于锥变性的营养不良的特定策略（AIM 3）。 gnat2floxstop/gnat2creert2可编程模型将为正在进行的贡献提供一个平台 旨在增加锥体介导的营养不良的基因治疗后，旨在增加视觉功能的努力。 它还将允许我们解决几个引人注目的临床相关问题：是大脑的电路 足够的塑性从GNAT2突变引起的病理变化中恢复过来？有什么 之后没有回报，基因型的目的地反向野生型，无法挽救锥体？可以暂时 通过代谢重编程可以缓解基因疗法的障碍？ 综上所述，该提议可以肯定地1）定义限制介入疗法的因素； 2）验证a 锥体介导的视网膜变性的新型，可诱导的模型； 3）确定代谢重新编程是否 可以作为治疗永久性变性的有效，非基因的策略。