The RNA-activation platform to treat X-linked disease in a locus-specific manner.

RNA 激活平台以位点特异性方式治疗 X 连锁疾病。

• 批准号: 8737219
• 负责人: JEANNIE T LEE
• 金额: $ 30.58万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8737219
• 关键词: Antisense Oligonucleotides; Binding; Cells; Code; Complex; DNA Methylation; Deacetylation; Development; Disease; Disease model; Dosage Compensation (Genetics); Drug Design; Drug Industry; Duchenne muscular dystrophy; Elements; Epigenetic Process; Equilibrium; Female; Fragile X Syndrome; Gene Activation; Gene Expression; Gene Silencing; Generations; Genes; Heterochromatin; Histone H3; Histones; Human; Knowledge; Laboratories; Legal patent; Licensing; Link; Lysine; Malignant Neoplasms; Mammals; Mediating; Methodology; Methods; Methylation; MicroRNAs; Modality; Modeling; Modification; Mus; Mutation; Patients; Pharmacologic Substance; Plague; Polycomb; Positioning Attribute; Preclinical Testing; Process; Proteins; RNA; RNA Interference; Research; Rett Syndrome; Site; Small RNA; Specificity; Tail; Technology; Testing; Therapeutic; Tissues; Untranslated RNA; Work; X Chromosome; X Inactivation; gene repression; human disease; in vivo; innovation; interest; male; mouse model; next generation; pre-clinical; public health relevance; small molecule; success; tool; transcriptome sequencing

DESCRIPTION (provided by applicant): The proposed research will develop methods of upregulating X-linked genes through technical innovations applied to the original "RNA-activation" technology developed in my laboratory (RNA-a; patent pending, PCT- US2011-065939). If successful, this next-generation RNA-a methodology will have the potential to cure dozens of intractable X-linked diseases such as Rett syndrome, Fragile X syndrome, and Duchenne muscular dystrophy. In the pharmaceutical industry, current therapeutic strategies focus almost exclusively on protein and microRNA targets. Yet, lncRNAs confer a temporal and spatial specificity not possible with proteins and small RNAs, and disease-associated mutations occur more often in noncoding than coding space. Therapeutic strategies might be as effectively targeted at long noncoding RNAs (lncRNA). Indeed, designing drugs against site-specific cis-acting lncRNAs would circumvent the pleiotropic effects plaguing modalities that general activities of epigenetic complexes via small molecules. The first-generation RNA-a strategy reawakens epigenetically silenced genes by disrupting binding between Polycomb repressive complex 2 (PRC2) and the lncRNA that targets it to the locus of interest. The RNA-a technology achieves the opposite of RNAi and has proven efficacious for various disease genes in preclinical testing. However, X-linked disease genes have not been amenable to RNA-a, likely because of many additional layers of heterochromatin resulting from X- chromosome inactivation (XCI). XCI is the mechanism of dosage compensation in mammals in which one X- chromosome is silenced in female cells to balance gene expression with male cells. My laboratory has specialized in the study of XCI for the past 16 years and recently discovered X-linked elements involved in the spreading of XCI along the inactive X chromosome (Xi). From these discoveries, we hypothesize that it should be possible to selectively reawaken specific Xi loci, without reactivating most or all of the Xi. Indeed, reactivating the entire Xi would be undesirable, as we have shown that global X-reactivation results in cancer. Thus, treating X-linked diseases requires a locus-specific approach to gene activation. With newfound knowledge of how XCI spreads, the envisioned next-generation RNA-a technology has the potential to do this. Because of our expertise in XCI and RNA-a, my lab is in a unique position to further develop the RNA-a technology to unlock expression of Xi genes in a locus-specific manner. Herein we propose to achieve this by leveraging gene-specific control elements along the Xi to reactivate the model disease gene, MECP2.

描述（由申请人提供）：拟议的研究将通过应用于我实验室开发的原始“RNA激活”技术的技术创新来开发上调X连锁基因的方法（RNA-a；专利申请中，PCT-US2011-065939）。如果成功，这种下一代 RNA-a 方法将有可能治愈数十种棘手的 X 连锁疾病，如雷特综合征、脆性 X 综合征和杜氏肌营养不良症。在制药行业，当前的治疗策略几乎完全集中在蛋白质和 microRNA 靶标上。然而，lncRNA 具有蛋白质和小 RNA 不可能实现的时间和空间特异性，并且与疾病相关的突变在非编码空间中比在编码空间中更常见。治疗策略可能同样有效地针对长非编码 RNA (lncRNA)。事实上，设计针对位点特异性顺式作用lncRNA的药物将规避困扰表观遗传复合物通过小分子的一般活性的多效性效应。第一代 RNA-a 策略通过破坏 Polycomb 抑制复合物 2 (PRC2) 和将其靶向目标位点的 lncRNA 之间的结合来重新唤醒表观遗传沉默的基因。 RNA-a技术实现了与RNAi相反的效果，并已在临床前测试中证明对多种疾病基因有效。然而，X连锁疾病基因不适合RNA-a，可能是因为X染色体失活（XCI）产生了许多额外的异染色质层。 XCI是哺乳动物中的剂量补偿机制，其中雌性细胞中的一条X染色体被沉默，以平衡雄性细胞的基因表达。我的实验室在过去 16 年里专门研究 XCI，最近发现了 X 连锁元件，这些元件参与 XCI 沿着不活跃的 X 染色体 (Xi) 的传播。根据这些发现，我们假设应该有可能选择性地重新唤醒特定的 Xi 位点，而无需重新激活大部分或全部 Xi。事实上，重新激活整个 Xi 是不可取的，因为我们已经证明全局 X 重新激活会导致癌症。因此，治疗 X 连锁疾病需要采用位点特异性的基因激活方法。凭借对 XCI 如何传播的新知识，设想的下一代 RNA-a 技术有潜力做到这一点。由于我们在 XCI 和 RNA-a 方面的专业知识，我的实验室处于独特的地位，可以进一步开发 RNA-a 技术，以位点特异性方式解锁 Xi 基因的表达。在此，我们建议通过利用 Xi 上的基因特异性控制元件来重新激活模型疾病基因 MECP2 来实现这一目标。