Pathobiology and Treatment of the UBTF E210K Neuroregression Syndrome

UBTF E210K 神经退行综合征的病理学和治疗

基本信息

批准号：
10416149
负责人：
MARK S LEDOUX
金额：
$ 41.47万
依托单位：
UNIVERSITY OF MEMPHIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10416149
关键词：
Affect Affinity Amino Acids Animal Model Apoptosis Apoptotic Architecture Awareness Behavioral Binding Bioavailable Biogenesis Biological Biological Assay Biological Models Brain Cell Cycle Cell Cycle Progression Cell Differentiation process Cell Nucleolus Cell model Cells ChIP-seq Chromatin Chromatin Structure ClinVar Clinical Treatment Cognitive Cognitive deficits Complex DNA DNA Damage DNA Double Strand Break DNA Polymerase II DNA-Binding Proteins DNA-Directed RNA Polymerase Data Deterioration Disease Drosophila genus Embryo Exhibits Failure Fibroblasts Functional disorder Gene Expression Gene Silencing Genes Genetic Recombination Genetic Transcription Genetically Engineered Mouse Genome Genome Stability Goals HMGB Proteins Histones Human Label Maps Messenger RNA Metabolic Molecular Motor Mus Mutation Nature Nerve Degeneration Nervous System Physiology Neuraxis Neurologic Neurons Onset of illness Oral Pathway interactions Patients Polymerase Preclinical Testing Protein Isoforms Psoralens RNA Processing RNA, Ribosomal, 18S Recombinant DNA Recurrence Research Reverse Transcriptase Polymerase Chain Reaction Ribosomal RNA Ribosomes Role Site Stress Structure Switch Genes Syndrome Techniques Transcription Elongation Transcription Initiation Transgenic Organisms Up-Regulation cell type crosslink early childhood experimental study factor C genome-wide improved inhibitor/antagonist motor deficit motor disorder mouse model mutant nervous system disorder neuron loss pre-clinical promoter rRNA Genes transcription factor transcriptome sequencing

项目摘要

Recurrent de novo dominant mutations in UBTF (NM_014233.3:c.628G>A, p.Glu210Lys) were recently associated with a distinct neurological disorder dubbed the UBTF E210K neuroregression syndrome. UBTF exists as two major isoforms, UBTF1 and UBTF2. UBTF1 regulates ribosomal RNA (rRNA) transcription by RNA polymerase 1 (Pol I), whereas UBTF2 regulates mRNA transcription by RNA polymerase 2 (Pol II). UBTF c.628G>A results in the same amino acid change in both UBTF1 and UBTF2 (E210K). Disease onset is at 2.5 to 3 yrs and characterized by slow progression of global motor, cognitive and behavioral dysfunction. UBTF E210K modified binding to the rDNA promoter that affects formation of the preinitiation complex by the Pol1-specific TBP-complex SL1. Since both UBTF1 and UBTF2 contribute to DNA accessibility and genomic stability, the increased binding affinity of the UBTF E210K mutant to DNA could alter the interchange with histone chromatin that occurs when genes are switched off, potentially leaving the underlying DNA poorly protected. UBTF E210K fibroblasts show increased expression of pre-rRNA and 18S rRNA, nucleolar abnormalities, markedly increased numbers of DNA double-strand breaks (DSBs), defective cell-cycle progression, and apoptosis. Ubtf-/- is early embryonic lethal in mice and transgenic expression of mutant human UBTF E210K in Drosophila neurons is also lethal. There are no known disease-modifying treatments for this disorder. Furthermore, we have not yet (i) clarified the relationships among increase expression of rRNA, DNA DSBs and cell-cycle abnormalities identified in patient fibroblasts, (ii) determined the mechanistic connections between nucleolar dysfunction and non-nucleolar DNA damage, or (iii) fully characterized a UBTF E210K mammalian model system. Our overall goals are to understand the mechanisms by which UBTF E210K causes progressive cellular dysfunction and develop a treatment for this devastating disorder. We hypothesize that Ubtf+/E210K mice will exhibit progressive motor and cognitive deficits, nucleolar abnormalities and apoptotic neuronal loss in association with accumulated DNA damage. Next, we will determine the effects of UBTF E210K on rRNA transcription, RNA processing, and RNA Pol I transcription elongation. Integrated analysis of ChIP-Seq and RNA-Seq will be used to determine the effects of UBTF E210K on chromatin structure and genome-wide gene expression. We will quantify the effects of UBTF E210K on rDNA, non-rDNA, and nucleolar structure. Psoralen cross-linking will be used to determine the ratio of active to inactive genes. ChIP-Seq mapping of γH2A.X will be used to map sites of DNA damage at the genome-wide level. Finally, we will test pre-clinical treatment of UBTF E210K cellular and animal models with orally-bioavailable and central nervous system penetrant Pol I inhibitors.

UBTF 中复发的从头显性突变 (NM_014233.3:c.628G>A, p.Glu210Lys) 最近被发现与一种称为 UBTF E210K 神经退行综合征的独特神经系统疾病有关。 UBTF1 和 UBTF2 两种主要同工型存在，UBTF1 通过调节核糖体 RNA (rRNA) 转录。 RNA 聚合酶 1 (Pol I)，而 UBTF2 通过 RNA 聚合酶 2 (Pol II) 调节 mRNA 转录。 UBTF c.628G>A 导致 UBTF1 和 UBTF2 (E210K) 中相同的氨基酸变化。 2.5 至 3 岁时，其特征是整体运动、认知和行为功能障碍进展缓慢。 UBTF E210K 修饰了与 rDNA 启动子的结合，从而影响了预起始复合物的形成 Pol1 特异性 TBP 复合物 SL1 因为 UBTF1 和 UBTF2 都有助于 DNA 可及性和基因组。稳定性，UBTF E210K 突变体与 DNA 的结合亲和力增加可能会改变与当基因关闭时发生的组蛋白染色质，可能会导致底层 DNA 质量不佳 UBTF E210K 成纤维细胞显示前 rRNA 和 18S rRNA 的表达增加，核仁。异常、DNA 双链断裂 (DSB) 数量显着增加、细胞周期缺陷 Ubtf-/-在小鼠中是早期胚胎致死的，并且是突变体的转基因表达。果蝇神经元中的人类 UBTF E210K 也是致命的。目前尚无已知的缓解疾病的治疗方法。此外，我们尚未阐明 (i) 表达增加之间的关系。在患者成纤维细胞中鉴定出 rRNA、DNA DSB 和细胞周期异常，(ii) 确定了机制核仁功能障碍和非核仁 DNA 损伤之间的联系，或 (iii) 充分表征 UBTF E210K 哺乳动物模型系统。我们的总体目标是了解 UBTF 的机制。 E210K 会导致进行性细胞功能障碍，并开发出针对这种破坏性疾病的治疗方法。 Ubtf+/E210K 小鼠将表现出进行性运动和认知缺陷、核仁异常以及与累积的 DNA 损伤相关的细胞凋亡神经元损失接下来，我们将确定其影响。 UBTF E210K 对 rRNA 转录、RNA 加工和 RNA Pol I 转录集成延伸的影响。 ChIP-Seq 和 RNA-Seq 分析将用于确定 UBTF E210K 对染色质的影响我们将量化 UBTF E210K 对 rDNA、非 rDNA、和核仁结构将用于确定活性基因与非活性基因的比率。 γH2A.X 的 ChIP-Seq 作图将用于在全基因组水平上绘制 DNA 损伤位点图谱。将测试具有口服生物利用度和中枢活性的 UBTF E210K 细胞和动物模型的临床前治疗神经系统渗透性 Pol I 抑制剂。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

HMG-boxes, ribosomopathies and neurodegenerative disease.

DOI：
10.3389/fgene.2023.1225832
发表时间：
2023
期刊：
FRONTIERS IN GENETICS
影响因子：
3.7
作者：
Moss, Tom;LeDoux, Mark S.;Crane-Robinson, Colyn
通讯作者：
Crane-Robinson, Colyn

Ribosomal DNA promoter recognition is determined in vivo by cooperation between UBTF1 and SL1 and is compromised in the UBTF-E210K neuroregression syndrome.

DOI：
10.1371/journal.pgen.1009644
发表时间：
2022-03
期刊：
PLoS genetics
影响因子：
4.5
作者：
Tremblay MG;Sibai DS;Valère M;Mars JC;Lessard F;Hori RT;Khan MM;Stefanovsky VY;LeDoux MS;Moss T
通讯作者：
Moss T

Presynaptic PRRT2 Deficiency Causes Cerebellar Dysfunction and Paroxysmal Kinesigenic Dyskinesia.

DOI：
10.1016/j.neuroscience.2020.08.034
发表时间：
2020-11-10
期刊：
NEUROSCIENCE
影响因子：
3.3
作者：
Calame, Dylan J.;Xiao, Jianfeng;Khan, Mohammad Moshahid;Hollingsworth, T. J.;Xue, Yi;Person, Abigail L.;LeDoux, Mark S.
通讯作者：
LeDoux, Mark S.

No Neuron Operates in Isolation.

没有神经元是孤立运作的。