Endogenous regulation of huntingtin expression as a therapeutic target for Huntington's disease

亨廷顿蛋白表达的内源调节作为亨廷顿病的治疗靶点

• 批准号: 10214706
• 负责人: RUSSELL L MARGOLIS
• 金额: $ 47.09万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214706
• 关键词: Adult; Alleles; Antibodies; Attention; Brain; Brain-Derived Neurotrophic Factor; CAG repeat; Cell model; Cells; Cessation of life; Chromatin; Chromatin Structure; Clinical; Data; Development; Disease; Disease Progression; Effectiveness; Exons; Face; Funding; Future; Genes; Huntington Disease; Huntington gene; Huntington protein; Individual; Inherited; Lead; Mediating; Methods; Molecular; Molecular Biology; Mutation; Neurodegenerative Disorders; Oligonucleotides; Onset of illness; Pathogenesis; Pathogenicity; Pathway interactions; Phase; Pilot Projects; Promoter Regions; Property; Proteins; Protocols documentation; RNA Splicing; Regulation; Research; Role; Series; Small Interfering RNA; Specificity; Structure; Testing; Therapeutic; Therapeutic Agents; Toxic effect; Transcript; Variant; Withdrawal; Work; base; behavioral phenotyping; brain cell; chromatin remodeling; experimental study; high throughput screening; human disease; induced pluripotent stem cell; knock-down; lymphoblast; mouse model; mutant; neurotoxic; new therapeutic target; novel; overexpression; prevent; programs; promoter; protein expression; small molecule; therapeutic target; tool; validation studies

Summary. Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder, characterized by relentless progression to death ~20 years after disease onset. HD is caused by an expanded CAG repeat in exon 1 of the huntingtin (HTT) gene. Disease pathogenesis is largely a result of expression of the mutant transcript and protein, which have neurotoxic properties. Suppressing the expression of the mutant allele is therefore a promising therapeutic approach, thus far pursued with antibody, oligonucleotide and siRNA strategies. As with any knockdown approach, especially in the CNS, problems of delivery, reversibility, and off-target effects using these methods remain problematic. Surprisingly, relatively little is known about the regulation of the HD locus, and in particular on the mechanisms that regulate HTT expression. We have recently discovered a gene on the strand antisense to HTT at the HD locus, which we have termed huntingtin antisense (HTT-AS). We have demonstrated that increasing expression of HTT-AS decreases expression of HTT, and that HTT-AS expression can be manipulated using small molecules. We hypothesize that HTT-AS itself, and the components that regulate its expression and interaction with HTT, will provide novel therapeutic targets for suppression of HTT expression and hence treatment of HD. A corollary to this hypothesis is that a better understanding of the HD locus, in this case the role of HTT-AS, will be critical in the interpretation of any HTT suppression strategy. We will test this hypothesis with a series of experiments organized into three specific aims, each built on compelling preliminary data. In Aim 1, we will determine if additional HTT-AS exons, splice variants, and promoters exist, determine the effect of repeat expansion on HTT-AS expression, and identify protein factors that regulate HTT-AS promoter activity. In Aim 2, we will determine the mechanisms by which HTT-AS suppresses HTT, including the quantitative effect of different HTT-AS transcripts on the suppression of HTT, and the role of chromatin remodeling on HTT expression. In Aim 3, we will collaborate with NCATS and CHDI to perform a large scale high throughput screen to find and characterize compounds that decrease expression of HTT by specifically increasing expression of HTT-AS. Selected compounds will be validated in cell and mouse models.

概括。 亨廷顿氏病（HD）是一种常染色体显性神经退行性疾病，其特征是 疾病发作后20年无情地进展。高清是由扩展的CAG引起的 在Huntingtin（HTT）基因的外显子1中重复。疾病发病机理在很大程度上是表达的结果 突变转录本和具有神经毒性特性的蛋白质。抑制表达 因此，突变等位基因是一种有希望的治疗方法 寡核苷酸和siRNA策略。与任何敲低方法一样，尤其是在中枢神经系统中 使用这些方法的交付，可逆性和非目标效应的问题仍然有问题。 令人惊讶的是，关于高清基因座的调节，尤其是对 调节HTT表达的机制。我们最近在链上发现了一个基因 在高清基因座上对HTT的反义，我们称之为Huntingtin反义（HTT-AS）。我们有 证明htt-as的表达增加会降低htt的表达，并且htt-as 可以使用小分子来操纵表达。我们假设这是HTT-本身，以及 调节其表达和与HTT相互作用的组件将提供新的治疗靶标 用于抑制HTT表达并因此治疗HD。这一假设的必然是 更好地理解高清基因座，在这种情况下，HTT-AS的作用将至关重要 解释任何HTT抑制策略。我们将通过一系列实验检验该假设 组织成三个特定目标，每个目标都建立在引人注目的初步数据上。在AIM 1中，我们将 确定是否存在其他HTT-AS外显子，剪接变体和启动子，确定 在HTT-AS表达上重复扩展，并确定调节HTT-AS启动子的蛋白质因子 活动。在AIM 2中，我们将确定HTT-AS抑制HTT的机制，包括 不同HTT-AS转录本对HTT抑制的定量效应和染色质的作用 重塑HTT表达。在AIM 3中，我们将与NCAT和CHDI合作，以执行大型 缩放高吞吐量屏幕以查找和表征化合物，以减少HTT的表达 特别增加了htt-as的表达。选定的化合物将在细胞和鼠标中进行验证 型号。

项目成果

A high-throughput screening to identify small molecules that suppress huntingtin promoter activity or activate huntingtin-antisense promoter activity.

• DOI: 10.1038/s41598-021-85279-2
• 发表时间: 2021-03-17
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Khaled HG;Feng H;Hu X;Sun X;Zheng W;Li PP;Rudnicki DD;Ye W;Chen YC;Southall N;Marugan J;Ross CA;Ferrer M;Henderson MJ;Margolis RL
• 通讯作者: Margolis RL

Use of single guided Cas9 nickase to facilitate precise and efficient genome editing in human iPSCs.

• DOI: 10.1038/s41598-021-89312-2
• 发表时间: 2021-05-10
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Li PP;Margolis RL
• 通讯作者: Margolis RL