Childhood-onset hypomyelinating leukodystrophy and the multi-tRNA synthetase complex
儿童期发病的低髓鞘性脑白质营养不良和多 tRNA 合成酶复合物
基本信息
- 批准号:10582441
- 负责人:
- 金额:$ 62.66万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-12-15 至 2027-11-30
- 项目状态:未结题
- 来源:
- 关键词:AffectAmino AcidsAmino Acyl-tRNA SynthetasesAnticodonAtaxiaAutomobile DrivingBehavioralBindingCRISPR/Cas technologyCell NucleusCellsCentral Nervous SystemCentral Nervous System DiseasesCerebellar DiseasesCerebrumChildChildhoodClinicalCodeCognitiveComplexCytoplasmDefectDevelopmental Delay DisordersDisparateDistantElementsEtiologyExhibitsFunctional disorderGait abnormalityGenesGenetic CodeGlutamatesGrowthHeterozygoteHuman InbreedingImpaired cognitionIntellectual functioning disabilityKnock-in MouseLamin Type ALettersLife ExpectancyLigaseLigationMediatingMessenger RNAMethylationMicrocephalyMolecularMotorMusMutant Strains MiceMutateMutationMyelinNeural ConductionNeurodegenerative DisordersNeurologicNeurologic DeficitNomenclatureNuclearNuclear ExportOligodendrogliaParentsPathologicPathologic NystagmusPathologyPathway interactionsPeripheral NervesPhenotypePoint MutationPoly(ADP-ribose) PolymerasesPositioning AttributePost-Transcriptional RegulationProlineProtein BiosynthesisProteinsRecombinantsRepressionRoleSiblingsSpecificitySystemTestingTherapeuticTherapeutic InterventionTrans-ActivatorsTranscriptTransfer RNATransfer RNA AminoacylationTranslationsVariantadverse outcomebrain magnetic resonance imagingcell typecurative treatmentsearly adolescenceexome sequencingexperimental studyglutamic acid-tRNAglutamyl-prolyl-tRNA synthetasein vivoin vivo Modelknock-downleukodystrophylymphoblastoid cell linemotor impairmentmouse modelmutantmyelinationnoveloligodendrocyte precursorpreclinical evaluationprecursor cellprenatalprogramsproline-tRNAprotein expressionsingle nucleus RNA-sequencingsmall hairpin RNAspasticitytherapeutic RNAtranscriptomicsvirtual
项目摘要
Project Summary/Abstract
Child-onset hypomyelinating leukodystrophy (HLD) is a genetically heterogeneous group of neurodegenerative
diseases characterized by reduced cerebral myelin formation. Clinical features include cognitive as well as
motor impairment appearing in childhood. There are no curative treatments. Our collaborator, Dr. Grace Yoon,
evaluated two siblings presenting with severe neurological deficit and a shared phenotype consisting of global
developmental delay and intellectual disability, prenatal onset undergrowth and microcephaly, rotatory
nystagmus, ataxia and progressive gait disturbance/spasticity, and hypomyelination on brain MRI. Whole
exome sequencing revealed homozygosity of a c.4444C>A mutation in the EPRS1 gene. The mutation is in
the coding sequence (cds) of the gene causing a Pro-to-Thr point mutation at aa position 1482. The EPRS1
gene encodes the bifunctional, glutamyl-prolyl tRNA synthetase (EPRS1) that resides in the cytoplasmic multi-
tRNA synthetase complex (MSC). The P1482T mutation is located near the C-terminus of the protein in a
region of the Pro synthetase outside the catalytic or anti-codon recognition domains. The specific activity of
recombinant P1482T mutant EPRS1 is indistinguishable from wild-type. In contrast, EPRS1 protein expression
in immortalized lymphoblastoid cell lines (LCL) from affected siblings is about 20% of that in unaffected
controls. EPRS1 mRNA levels are identical in siblings and controls indicating post-transcriptional regulation.
Our preliminary studies show a dual mechanism determining diminished EPRS1 level in affected LCLs,
namely, decreased nuclear export of mutant mRNA, followed by decreased cytoplasmic translation. We show
that decreased EPRS1 expression causes release of other MSC constituents. Remarkably, suppression of
several other MSC components causally implicated in HLD also induce MSC constituent release, suggesting a
common etiology of an entire class of HLDs. We hypothesize that inefficient nuclear export and translation of
c.4444C>A EPRS1 mRNA reduces MSC-bound EPRS1, causing release of MSC constituents and driving the
HLD phenotype. Likewise, defects in genes encoding other MSC constituents share a common pathway and
etiology of HLD. We will test this hypothesis by pursuing these Specific Aims: In Aim 1 we will elucidate
molecular mechanisms underlying reduced expression of EPRS1P1482T variant, focusing on the role of m6A
methylation. Aim 2 determines the HLD-related phenotype of our newly generated, genetically-modified
Eprs1P1482T mice. In Aim 3 we will investigate effect of deficiency of MSC constituents on MSC integrity and
adverse consequences in myelinating oligodendrocytes. A CNS-specific transcriptomic analysis will be done by
single-nucleus RNA-sequencing. Completion of these studies will elucidate a unique mechanism of gene
dysregulation that induces HLD, and will provide a unique mouse model of HLD that will permit detailed in vivo
analysis of the cellular defects in HLD. Importantly, these results suggest the possibility of specific, RNA-based
therapeutic intervention to rescue translation of the variant EPRS1 mRNA and rescue the neurologic defect.
项目概要/摘要
儿童发病的低髓鞘性脑白质营养不良(HLD)是一组遗传异质性神经退行性疾病
以脑髓磷脂形成减少为特征的疾病。临床特征包括认知以及
儿童时期出现的运动障碍。没有治愈方法。我们的合作者 Grace Yoon 博士,
评估了两个患有严重神经缺陷和共同表型的兄弟姐妹
发育迟缓和智力障碍、产前生长发育不良和小头畸形、旋转
眼球震颤、共济失调和进行性步态障碍/痉挛,以及脑 MRI 上的髓鞘形成不足。所有的
外显子组测序揭示了 EPRS1 基因中 c.4444C>A 突变的纯合性。突变是在
导致第 1482 位氨基酸发生 Pro-to-Thr 点突变的基因的编码序列 (cds)。EPRS1
该基因编码双功能谷氨酰脯氨酰 tRNA 合成酶 (EPRS1),该酶位于细胞质多
tRNA 合成酶复合物 (MSC)。 P1482T 突变位于蛋白质 C 末端附近
Pro 合成酶的催化域或反密码子识别域之外的区域。具体活动为
重组 P1482T 突变体 EPRS1 与野生型没有区别。相反,EPRS1蛋白表达
来自受影响兄弟姐妹的永生化淋巴母细胞系 (LCL) 约为未受影响的 20%
控制。兄弟姐妹和对照中的 EPRS1 mRNA 水平相同,表明转录后调节。
我们的初步研究表明,双重机制决定了受影响 LCL 中 EPRS1 水平的降低,
也就是说,突变体 mRNA 的核输出减少,随后细胞质翻译减少。我们展示
EPRS1 表达减少会导致其他 MSC 成分的释放。值得注意的是,抑制
与 HLD 有因果关系的其他几种 MSC 成分也会诱导 MSC 成分释放,这表明
整个 HLD 类别的共同病因。我们假设低效的核输出和翻译
c.4444C>A EPRS1 mRNA 减少 MSC 结合的 EPRS1,导致 MSC 成分释放并驱动
HLD表型。同样,编码其他 MSC 成分的基因缺陷具有共同的途径,并且
HLD 的病因学。我们将通过追求这些具体目标来检验这一假设:在目标 1 中,我们将阐明
EPRS1P1482T 变体表达减少的分子机制,重点关注 m6A 的作用
甲基化。目标 2 确定我们新生成的转基因细胞的 HLD 相关表型
Eprs1P1482T 小鼠。在目标 3 中,我们将研究 MSC 成分缺乏对 MSC 完整性的影响和
髓鞘少突胶质细胞的不良后果。 CNS 特异性转录组分析将由
单核RNA测序。这些研究的完成将阐明基因的独特机制
诱导 HLD 的失调,并将提供独特的 HLD 小鼠模型,该模型将允许详细的体内研究
HLD 中的细胞缺陷分析。重要的是,这些结果表明基于 RNA 的特异性研究的可能性
治疗干预以挽救变异 EPRS1 mRNA 的翻译并挽救神经系统缺陷。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Ranjan Dutta其他文献
Ranjan Dutta的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Ranjan Dutta', 18)}}的其他基金
Molecular correlates of sub-regional thalamic degeneration in multiple sclerosis
多发性硬化症亚区域丘脑变性的分子相关性
- 批准号:
10449474 - 财政年份:2022
- 资助金额:
$ 62.66万 - 项目类别:
Understanding role of circadian disruption in pathogenesis of MS
了解昼夜节律紊乱在多发性硬化症发病机制中的作用
- 批准号:
10442857 - 财政年份:2022
- 资助金额:
$ 62.66万 - 项目类别:
Molecular correlates of sub-regional thalamic degeneration in multiple sclerosis
多发性硬化症亚区域丘脑变性的分子相关性
- 批准号:
10553206 - 财政年份:2022
- 资助金额:
$ 62.66万 - 项目类别:
Understanding role of circadian disruption in pathogenesis of MS
了解昼夜节律紊乱在多发性硬化症发病机制中的作用
- 批准号:
10574570 - 财政年份:2022
- 资助金额:
$ 62.66万 - 项目类别:
MicroRNAs as critical regulators of remyelination in Multiple Sclerosis
MicroRNA 作为多发性硬化症髓鞘再生的关键调节因子
- 批准号:
9272452 - 财政年份:2016
- 资助金额:
$ 62.66万 - 项目类别:
相似海外基金
Double Incorporation of Non-Canonical Amino Acids in an Animal and its Application for Precise and Independent Optical Control of Two Target Genes
动物体内非规范氨基酸的双重掺入及其在两个靶基因精确独立光学控制中的应用
- 批准号:
BB/Y006380/1 - 财政年份:2024
- 资助金额:
$ 62.66万 - 项目类别:
Research Grant
Quantifying L-amino acids in Ryugu to constrain the source of L-amino acids in life on Earth
量化 Ryugu 中的 L-氨基酸以限制地球生命中 L-氨基酸的来源
- 批准号:
24K17112 - 财政年份:2024
- 资助金额:
$ 62.66万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
Molecular recognition and enantioselective reaction of amino acids
氨基酸的分子识别和对映选择性反应
- 批准号:
23K04668 - 财政年份:2023
- 资助金额:
$ 62.66万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Basic research toward therapeutic strategies for stress-induced chronic pain with non-natural amino acids
非天然氨基酸治疗应激性慢性疼痛策略的基础研究
- 批准号:
23K06918 - 财政年份:2023
- 资助金额:
$ 62.66万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Molecular mechanisms how arrestins that modulate localization of glucose transporters are phosphorylated in response to amino acids
调节葡萄糖转运蛋白定位的抑制蛋白如何响应氨基酸而被磷酸化的分子机制
- 批准号:
23K05758 - 财政年份:2023
- 资助金额:
$ 62.66万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Design and Synthesis of Fluorescent Amino Acids: Novel Tools for Biological Imaging
荧光氨基酸的设计与合成:生物成像的新工具
- 批准号:
2888395 - 财政年份:2023
- 资助金额:
$ 62.66万 - 项目类别:
Studentship
Collaborative Research: RUI: Elucidating Design Rules for non-NRPS Incorporation of Amino Acids on Polyketide Scaffolds
合作研究:RUI:阐明聚酮化合物支架上非 NRPS 氨基酸掺入的设计规则
- 批准号:
2300890 - 财政年份:2023
- 资助金额:
$ 62.66万 - 项目类别:
Continuing Grant
Structurally engineered N-acyl amino acids for the treatment of NASH
用于治疗 NASH 的结构工程 N-酰基氨基酸
- 批准号:
10761044 - 财政年份:2023
- 资助金额:
$ 62.66万 - 项目类别:
Lifestyle, branched-chain amino acids, and cardiovascular risk factors: a randomized trial
生活方式、支链氨基酸和心血管危险因素:一项随机试验
- 批准号:
10728925 - 财政年份:2023
- 资助金额:
$ 62.66万 - 项目类别:
Single-molecule protein sequencing by barcoding of N-terminal amino acids
通过 N 端氨基酸条形码进行单分子蛋白质测序
- 批准号:
10757309 - 财政年份:2023
- 资助金额:
$ 62.66万 - 项目类别: