Identifying regulatory uORFs as a targetable axis for hereditary disease

识别调节性 uORF 作为遗传性疾病的靶向轴

• 批准号: 10797954
• 负责人: Yoseph Barash
• 金额: $ 7.64万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-23 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10797954
• 关键词: 5' Untranslated Regions; Affect; BMPR2 gene; Cells; Code; Data; Disease; Evolution; Exhibits; Gene Proteins; Genes; Genetic Databases; Genetic Diseases; Goals; Hereditary Disease; Heritability; Human; Initiator Codon; Intervention; Knowledge; Messenger RNA; Open Reading Frames; Output; Peptides; Proteins; Publishing; Regulatory Element; Terminator Codon; Therapeutic Intervention; Translations; Variant; Work; clinically relevant; disease phenotype; mouse model; novel; parent grant; preservation; protein expression; pulmonary arterial hypertension; targeted treatment; translational approach

Abstract (from parent grant) The broad goal of this proposal it to computationally identify, validate, then target for therapeutic intervention regulatory elements within the 5' untranslated regions (UTR) of protein coding genes, known as upstream open reading frames (uORFs). In so doing, we aim to modulate the protein output from selected genes, offering a novel, translational approach with broad potential. uORFs are segments of 5′UTR mRNA sequences that can initiate and terminate translation upstream of protein-coding (CDS) start codons. Rare uORFs have been shown to produce potentially functional micro-peptides, while others affect protein expression by tuning translation rates of downstream protein-coding sequences. Our recent work showed that uORFs exhibit strong negative selection for preserving their start and stop codons, but not the encoded sequence, favoring the notion that uORFs have been retained through evolution because of their cis regulatory effect on protein gene products. Using genetic databases, we showed that variants affecting uORF start/stop codons associate with disease phenotypes (PheWAS analysis) and validated their expected effect on the protein (but not RNA) levels of the downstream genes. Potential uORFs have been identified in ~50% of all human protein-coding genes, many of which harbor more than one uORF. While blocking some uORFs decreases CDS protein output, our unpublished data show that targeting uORFs can be used to increase CDS translation. As a potential case for increasing protein levels, we focused on heritable pulmonary arterial hypertension (PAH), a fatal condition with no current cure, most commonly driven by BMPR2 haploinsufficiency. By blocking a BMPR2 uORF, we were able to increase BMPR2 protein levels by up to 220%. Based on our published and unpublished results we propose an integrative set of computational and experimental approaches to systematically detect, prioritize, validate and target uORFs for therapeutic intervention, focusing on genes associated with haploinsufficiency and ASOs as an intervention.

摘要（来自父母赠款） 该建议的广泛目标是在计算上识别，验证，然后进行治疗目标 蛋白质编码基因的5'未翻译区域（UTR）内的干预调节元件， 被称为上游的开放式阅读帧（UORFS）。这样，我们的目标是调节蛋白质 来自选定基因的输出，提供具有广泛潜力的新颖的翻译方法。 UORF是5'UTR mRNA序列的段，可以启动和终止翻译 蛋白质编码（CD）起始密码子的上游。罕见的UORF已显示出产生 潜在的功能性微肽，而另一些则通过调整翻译影响蛋白质表达 下游蛋白质编码序列的速率。我们最近的工作表明UORFS展览 强烈的否定选择以保留其起始和停止密码子，但没有编码 序列，偏爱通过进化来保留UORF的观念 顺式调节对蛋白质基因产物的影响。使用遗传数据库，我们显示了变体 影响UORF的开始/停止密码子与疾病表型（PHEWAS分析）和 验证了其对下游基因蛋白质（但不是RNA）水平的预期作用。 在所有人类蛋白质编码基因的约50％中已经确定了潜在的UORF，其中许多 藏有多个Uorf。在阻止某些UORF的同时降低了CDS蛋白的输出，我们 未发表的数据表明，靶向UORF可用于增加CDS翻译。作为 蛋白质水平升高的潜在病例，我们专注于遗传性肺动脉 高血压（PAH），一种致命状况，没有电流治愈，最常见的是BMPR2驱动 单倍不足。通过阻止BMPR2 UORF，我们能够提高BMPR2蛋白水平 高达220％。根据我们已发表和未发表的结果，我们提出了一组集成的集合 系统检测，优先级，验证和实验方法的计算和实验方法 针对治疗干预的目标UORF，重点关注与单倍不足有关的基因 和ASO作为干预措施。