Genetic Analysis of Neuronal Hypoxia Resistance
神经元耐缺氧的遗传分析
基本信息
- 批准号:10835277
- 负责人:
- 金额:$ 10.44万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2012
- 资助国家:美国
- 起止时间:2012-04-15 至 2025-07-31
- 项目状态:未结题
- 来源:
- 关键词:AddressAdministrative SupplementAntioxidantsAutophagocytosisBindingBiologicalCOVID-19CRISPR/Cas technologyCaenorhabditis elegansCellsCerebral PalsyDiseaseDrug Metabolic DetoxicationEnhancersEnvironmentEnzymesEquipmentEquipment FailureEtiologyFree RadicalsFundingGenesGeneticGenetic Enhancer ElementGluconeogenesisHydroxylationHypoxiaHypoxia Inducible FactorIschemic StrokeLifeMalignant NeoplasmsMetabolicMetabolismMitochondriaMyocardial InfarctionNerve DegenerationNeuronal HypoxiaNeuronsOrganismOxidative StressPathway interactionsPlayProcollagen-Proline DioxygenaseProductionPulmonary HypertensionReporterReproducibilityResistanceRoleSourceStressTestingTissuesTranscriptional Regulationcombatdeprivationdisorder preventionexperimental studygenetic analysishuman diseasehypoxia inducible factor 1in vivomodel organismnovelprotective pathwayreceptorresponsetherapeutic targettissue culturetranscription factortranscriptome
项目摘要
PROJECT SUMMARY
Hypoxia (O2 deprivation) plays a central role in diverse human diseases, including ischemic stroke, myocardial
infarction, pulmonary hypertension, Cerebral Palsy, COVID-19, and cancer. Metazoans respond to hypoxia by
employing the conserved hypoxia response pathway. The pathway senses O2 through a prolyl hydroxylase
(PHD) enzyme, which negatively regulates the transcription factor Hypoxia Inducible Factor α (HIFα). When
hypoxia ensues, PHD enzymes lack O2 to hydroxylate HIFα, resulting in HIFα stabilization and the
transcriptional regulation of multiple target genes that help the organism survive. While the HIFα pathway has
been well studied in tissue culture, a full understanding of how it operates in specific tissues (particularly
neurons) in vivo to provide tailored responses is needed. This proposal takes advantage of genetics and an
intact, isogenic model organism (C. elegans) that can thrive under hypoxia, and whose environment and
genetics can be controlled with fidelity and reproducibility. The overall premise of this proposal is that the
hypoxia response pathway pathway protects against hypoxic damage by (1) removing mitochondria through
mitophagy, which eliminates a source of oxidative stress, and by (2) mobilizing antioxidant metabolism, which
detoxifies free radicals during hypoxia and reoxygenation.
We hypothesize that HIF-1 promotes this metabolic reprograming by binding an enhancer sequence
and activating the expression of the PEP carboxykinase pck-1, a key enzyme for moving metabolites through
gluconeogenesis. Aim 1 tests this hypothesis by using CRISPR/Cas9 editing to remove this enhancer, then
testing for the effects on HIF-1 binding, pck-1 and global gene expression, metabolism, oxidative stress
resistance, neurodegeneration, and hypoxia survival.
Preliminary cell biological approaches with a genetically encoded fluorescent reporter for mitophagy
suggest that HIF-1 promotes mitophagy. We hypothesize that HIF-1 promotes mitophagy by binding
enhancer sequences and activating the expression of the mitophagy receptors fndc-1 and dct-1. Aim 2 tests
this hypothesis by using CRISPR/Cas9 editing to remove these enhancers, then testing for the effects on HIF-
1 binding, global gene expression, mitophagy and bulk autophagy, metabolism, oxidative stress resistance,
neurodegeneration, and hypoxia survival.
Recently, our media sterilizer and automated Petri dish pourer reached the end of its life. This large
equipment failure is inhibiting our progress towards both aims, as this fundamental equipment is essential for
every experiment we conduct. An administrative supplement is requested to replace this equipment and
restore our progress towards understanding how the hypoxia response pathway operates. A better
understanding of the pathway will provide therapeutic targets for diseases associated with hypoxia.
项目概要
缺氧(缺氧)在多种人类疾病中发挥着核心作用,包括缺血性中风、心肌病
梗塞、肺动脉高压、脑瘫、COVID-19 和癌症。后生动物对缺氧的反应是
采用保守的缺氧反应途径。该途径通过脯氨酰羟化酶感知 O2
(PHD) 酶,负向调节转录因子缺氧诱导因子 α (HIFα)。什么时候
当缺氧发生时,PHD 酶缺乏 O2 来羟基化 HIFα,从而导致 HIFα 稳定和
帮助生物体生存的多个靶基因的转录调控。虽然 HIFα 通路有
在组织培养中得到了充分的研究,充分了解它如何在特定组织(特别是
神经元)在体内提供定制的反应是必要的。该提议利用了遗传学和
完整的等基因模式生物(秀丽隐杆线虫),可以在缺氧下茁壮成长,其环境和
可以以保真度和再现性控制遗传学。该提案的总体前提是
缺氧反应途径通过(1)去除线粒体来防止缺氧损伤
线粒体自噬,消除氧化应激的来源,并通过(2)调动抗氧化代谢,
在缺氧和复氧过程中解毒自由基。
我们假设 HIF-1 通过结合增强子序列来促进这种代谢重编程
并激活 PEP 羧激酶 pck-1 的表达,这是一种将代谢物转移到体内的关键酶
糖异生。目标 1 通过使用 CRISPR/Cas9 编辑删除该增强子来测试该假设,然后
测试对 HIF-1 结合、pck-1 和整体基因表达、代谢、氧化应激的影响
抵抗力、神经退行性变和缺氧生存。
使用基因编码荧光报告基因进行线粒体自噬的初步细胞生物学方法
表明 HIF-1 促进线粒体自噬。我们假设 HIF-1 通过结合促进线粒体自噬
增强子序列并激活线粒体自噬受体 fndc-1 和 dct-1 的表达。目标 2 测试
这个假设是通过使用 CRISPR/Cas9 编辑去除这些增强子,然后测试对 HIF 的影响
1 结合、全局基因表达、线粒体自噬和大量自噬、代谢、氧化应激抵抗、
神经退行性变和缺氧生存。
最近,我们的培养基灭菌器和自动培养皿倾倒器达到了其使用寿命。这个大
设备故障阻碍了我们实现这两个目标的进展,因为这种基本设备对于
我们进行的每一个实验。需要行政补充来更换该设备,并且
恢复我们在理解缺氧反应途径如何运作方面的进展。更好的
了解该途径将为缺氧相关疾病提供治疗靶点。
项目成果
期刊论文数量(12)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
The hypoxia response pathway promotes PEP carboxykinase and gluconeogenesis in C. elegans.
- DOI:10.1038/s41467-022-33849-x
- 发表时间:2022-10-18
- 期刊:
- 影响因子:16.6
- 作者:
- 通讯作者:
Anoxia-reoxygenation regulates mitochondrial dynamics through the hypoxia response pathway, SKN-1/Nrf, and stomatin-like protein STL-1/SLP-2.
- DOI:10.1371/journal.pgen.1004063
- 发表时间:2013
- 期刊:
- 影响因子:4.5
- 作者:Ghose P;Park EC;Tabakin A;Salazar-Vasquez N;Rongo C
- 通讯作者:Rongo C
Going mobile: AMPA receptors move synapse to synapse in vivo.
- DOI:10.1016/j.neuron.2013.11.031
- 发表时间:2013-12-18
- 期刊:
- 影响因子:16.2
- 作者:Rongo C
- 通讯作者:Rongo C
RAB-6.1 and RAB-6.2 Promote Retrograde Transport in C. elegans.
RAB-6.1 和 RAB-6.2 促进线虫逆行运输。
- DOI:10.1371/journal.pone.0149314
- 发表时间:2016
- 期刊:
- 影响因子:3.7
- 作者:Zhang,Donglei;Dubey,Jyoti;Koushika,SandhyaP;Rongo,Christopher
- 通讯作者:Rongo,Christopher
RAB-6.2 and the retromer regulate glutamate receptor recycling through a retrograde pathway.
- DOI:10.1083/jcb.201104141
- 发表时间:2012-01-09
- 期刊:
- 影响因子:0
- 作者:Zhang D;Isack NR;Glodowski DR;Liu J;Chen CC;Xu XZ;Grant BD;Rongo C
- 通讯作者:Rongo C
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Christopher G Rongo其他文献
Christopher G Rongo的其他文献
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{{ truncateString('Christopher G Rongo', 18)}}的其他基金
Multi-Omic Analysis of BMP-Insulin Signaling Crosstalk in Lipid Metabolism during Aging
衰老过程中脂质代谢中 BMP-胰岛素信号串扰的多组学分析
- 批准号:
10351581 - 财政年份:2022
- 资助金额:
$ 10.44万 - 项目类别:
Multi-Omic Analysis of BMP-Insulin Signaling Crosstalk in Lipid Metabolism during Aging
衰老过程中脂质代谢中 BMP-胰岛素信号串扰的多组学分析
- 批准号:
10553134 - 财政年份:2022
- 资助金额:
$ 10.44万 - 项目类别:
Genetic Analysis of Neuronal Hypoxic Stress Resistance
神经元耐缺氧应激的遗传分析
- 批准号:
9753252 - 财政年份:2012
- 资助金额:
$ 10.44万 - 项目类别:
Genetics Analysis of Neuronal Hypoxic Stress Resistance
神经元耐缺氧应激的遗传学分析
- 批准号:
8650508 - 财政年份:2012
- 资助金额:
$ 10.44万 - 项目类别:
Genetic Analysis of Neuronal Hypoxic Stress Resistance
神经元耐缺氧应激的遗传分析
- 批准号:
9979647 - 财政年份:2012
- 资助金额:
$ 10.44万 - 项目类别:
Genetics Analysis of Neuronal Hypoxic Stress Resistance
神经元耐缺氧应激的遗传学分析
- 批准号:
8457043 - 财政年份:2012
- 资助金额:
$ 10.44万 - 项目类别:
Genetics Analysis of Neuronal Hypoxic Stress Resistance
神经元耐缺氧应激的遗传学分析
- 批准号:
8629773 - 财政年份:2012
- 资助金额:
$ 10.44万 - 项目类别:
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