NEURODEVELOPMENTAL FUNCTION OF HCFC1
HCFC1 的神经发育功能
基本信息
- 批准号:10541351
- 负责人:
- 金额:$ 4.14万
- 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:2022-07-01 至 2023-06-30
- 项目状态:已结题
- 来源:
- 关键词:AKT inhibitionAchievementAdultAllelesAnimalsAwardBehaviorBehavioralBindingBrainBrain DiseasesCell CountCell Differentiation processCell ProliferationCell SeparationCobalaminDataDensitometryDevelopmentDiseaseEpilepsyFRAP1 geneFellowshipFishesFlow CytometryFluorescence-Activated Cell SortingFutureGene ExpressionGenesGenetic TranscriptionGoalsHereditary DiseaseHumanImageImpairmentIn VitroIntellectual functioning disabilityIntractable EpilepsyKnowledgeLabelLarvaLinkManuscriptsMetabolismMolecularMolecular BiologyMusMutationNeurodevelopmental DisorderNeurogliaNeurologicNeuronsNeurosciencesNeurosphereNonsense MutationOrthologous GenePathway interactionsPentylenetetrazolePhasePhenotypePhosphorylationPostdoctoral FellowPredispositionPreparationProliferatingProteinsProteomicsProto-Oncogene Proteins c-aktReporterResearchRoleS phaseScienceSeizuresSignal TransductionSignaling MoleculeSirolimusSyndromeTechniquesTestingTimeTrainingTraining ProgramsTransgenic OrganismsWestern BlottingZebrafishbrain abnormalitiescell typechromatin immunoprecipitationcongenital anomalycritical periodhost cell factor C1in vivoinhibitorknock-downloss of function mutationmTOR InhibitormTOR inhibitionmotor disordermouse modelmutantnerve stem cellneuron developmentnovelpreventprofessorpromotersingle-cell RNA sequencingskillsstem cellstherapeutic targettranscription factortranscriptomicsupstream kinase
项目摘要
Project Summary
Mutation of HCFC1 causes a multiple congenital anomaly syndrome characterized by inborn errors of cobalamin
metabolism, intractable epilepsy, intellectual disability, and motor dysfunction. Despite an implication for HCFC1
in these neurological impairments, a mechanism describing the function of HCFC1 during brain development
has not been completely elucidated. HCFC1 encodes for a transcriptional co-factor protein known to regulate
cellular proliferation of various progenitor cells including neural precursor cells (NPC). NPCs undergo rapid
expansion during early brain development and differentiate into all the major cell types in the brain (i.e. neurons,
glia). To begin to elucidate a putative mechanism for HCFC1 in NPC expansion, we created the Co60 allele
which introduces a loss of function mutation in the zebrafish hcfc1a ortholog. Through immunohistochemical
labeling and cell counts, we demonstrated that heterozygous carriers of the Co60 allele (Co60/+) had increased
proliferation of NPCs. We next used transcriptomics of Co60/+ whole brain homogenates to reveal a 14-fold
increase in the expression of asxl1, a transcription factor critical for cell proliferation and activation of AKT
signaling. We found that inhibition of PI3K, an upstream activator of AKT, in Co60/+ mutants restored asxl1
dependent NPC over proliferation. Moreover, preliminary western blotting and densitometry analysis of our
mutants confirm hyperphosphorylation of AKT (Thr308). We next used chromatin immunoprecipitation to confirm
a direct binding of human HCFC1 to the zebrafish asxl1 promoter. Together, these findings indirectly link for the
first time hcfc1a function and asxl1 expression with AKT activation and NPC proliferation. What remains to be
understood is the level of AKT signaling in isolated NPCs derived from hcfc1a mutants and which AKT
downstream signaling molecules, like mTOR, regulate proliferation. Completion of this study will help to identify
novel molecular pathways that regulate brain development downstream of HCFC1 and pinpoint mechanisms as
to how its dysregulation contributes to neurodevelopmental disorders. During the F99 phase of this award, I seek
to gain working knowledge of techniques in molecular biology that include protein isolation, western
immunoblotting, fluorescence activated cell sorting (FACS), flow cytometry, advanced imaging, and behavioral
neuroscience. Additionally, I aim to refine my skills and obtain professional development in grantsmanship,
manuscript review and preparation, build and maintain my science network, and finalize my dissertation
research.
项目概要
HCFC1突变导致多种先天性异常综合征,其特征是钴胺素先天性缺陷
新陈代谢、顽固性癫痫、智力障碍和运动功能障碍。尽管对 HCFC1 有影响
在这些神经损伤中,描述了 HCFC1 在大脑发育过程中的功能的机制
尚未完全阐明。 HCFC1 编码已知可调节的转录辅助因子蛋白
包括神经前体细胞(NPC)在内的各种祖细胞的细胞增殖。 NPC 经历快速
在早期大脑发育过程中扩张并分化成大脑中的所有主要细胞类型(即神经元、
胶质细胞)。为了开始阐明 HCFC1 在 NPC 扩增中的假定机制,我们创建了 Co60 等位基因
这在斑马鱼 hcfc1a 直向同源物中引入了功能丧失突变。通过免疫组化
标记和细胞计数,我们证明 Co60 等位基因 (Co60/+) 的杂合携带者有所增加
NPC 的增殖。接下来,我们使用 Co60/+ 全脑匀浆的转录组学揭示了 14 倍
asxl1 的表达增加,asxl1 是一种对细胞增殖和 AKT 激活至关重要的转录因子
发信号。我们发现,在 Co60/+ 突变体中抑制 PI3K(AKT 的上游激活剂)可以恢复 asxl1
依赖NPC过度增殖。此外,我们的初步蛋白质印迹和光密度分析
突变体证实 AKT (Thr308) 过度磷酸化。接下来我们使用染色质免疫沉淀来确认
人 HCFC1 与斑马鱼 asxl1 启动子的直接结合。总之,这些发现间接链接到
首次 hcfc1a 功能和 asxl1 表达与 AKT 激活和 NPC 增殖有关。还剩下什么
了解了源自 hcfc1a 突变体的分离 NPC 中 AKT 信号传导的水平,以及 AKT 信号传导水平
下游信号分子,如 mTOR,调节增殖。完成本研究将有助于确定
调节 HCFC1 下游大脑发育的新分子途径,并查明机制
其失调如何导致神经发育障碍。在这个奖项的 F99 阶段,我寻求
获得分子生物学技术的工作知识,包括蛋白质分离、西方
免疫印迹、荧光激活细胞分选 (FACS)、流式细胞术、高级成像和行为学
神经科学。此外,我的目标是提高我的技能并获得资助方面的专业发展,
稿件审查和准备,建立和维护我的科学网络,并完成我的论文
研究。
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 }}
Victoria L Castro其他文献
Victoria L Castro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('Victoria L Castro', 18)}}的其他基金
Gene regulatory mechanisms governed by the ASXL1/HCF1/OGT complex during neurogenesis
神经发生过程中 ASXL1/HCF1/OGT 复合物控制的基因调控机制
- 批准号:
10794902 - 财政年份:2022
- 资助金额:
$ 4.14万 - 项目类别:
相似海外基金
Collaborative Research: Using Adaptive Lessons to Enhance Motivation, Cognitive Engagement, And Achievement Through Equitable Classroom Preparation
协作研究:通过公平的课堂准备,利用适应性课程来增强动机、认知参与和成就
- 批准号:
2335802 - 财政年份:2024
- 资助金额:
$ 4.14万 - 项目类别:
Standard Grant
Collaborative Research: Using Adaptive Lessons to Enhance Motivation, Cognitive Engagement, And Achievement Through Equitable Classroom Preparation
协作研究:通过公平的课堂准备,利用适应性课程来增强动机、认知参与和成就
- 批准号:
2335801 - 财政年份:2024
- 资助金额:
$ 4.14万 - 项目类别:
Standard Grant
A Longitudinal Study of the Relationship between Participation in a Comprehensive Exercise Program and Academic Achievement
参加综合锻炼计划与学业成绩之间关系的纵向研究
- 批准号:
24K14615 - 财政年份:2024
- 资助金额:
$ 4.14万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Collaborative Research: Characterizing Best Practices of Instructors who Have Narrowed Performance Gaps in Undergraduate Student Achievement in Introductory STEM Courses
合作研究:缩小本科生 STEM 入门课程成绩差距的讲师的最佳实践
- 批准号:
2420369 - 财政年份:2024
- 资助金额:
$ 4.14万 - 项目类别:
Standard Grant
Collaborative Research: Using Adaptive Lessons to Enhance Motivation, Cognitive Engagement, And Achievement Through Equitable Classroom Preparation
协作研究:通过公平的课堂准备,利用适应性课程来增强动机、认知参与和成就
- 批准号:
2335800 - 财政年份:2024
- 资助金额:
$ 4.14万 - 项目类别:
Standard Grant
WTG: Diffusion of Research on Supporting Mathematics Achievement for Youth with Disabilities through Twitter Translational Visual Abstracts
WTG:通过 Twitter 翻译视觉摘要传播支持残疾青少年数学成就的研究
- 批准号:
2244734 - 财政年份:2023
- 资助金额:
$ 4.14万 - 项目类别:
Standard Grant
The Impact of Emotional Experiences of Pride on Long-Term Goal Achievement Behaviors in Elite Athletes
骄傲的情感体验对优秀运动员长期目标实现行为的影响
- 批准号:
23K16740 - 财政年份:2023
- 资助金额:
$ 4.14万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
Meta-Analysis of the Instructional-Relational Model of Student Engagement and Math Achievement: A Moderation and Mediation Approach
学生参与度和数学成绩的教学关系模型的元分析:一种调节和中介方法
- 批准号:
2300738 - 财政年份:2023
- 资助金额:
$ 4.14万 - 项目类别:
Standard Grant
Improving maths achievement in children with speech, language, and communication needs through 'collaborative vocabulary teaching'
通过“协作词汇教学”提高有言语、语言和交流需求的儿童的数学成绩
- 批准号:
2890475 - 财政年份:2023
- 资助金额:
$ 4.14万 - 项目类别:
Studentship
HSI Institutional Transformation Project: Retention and Achievement for Introductory STEM English Learners (RAISE)
HSI 机构转型项目:STEM 英语入门学习者的保留和成就 (RAISE)
- 批准号:
2225178 - 财政年份:2023
- 资助金额:
$ 4.14万 - 项目类别:
Continuing Grant