Cholesterol regulation of smooth muscle BK channel proteins and consequent control of cerebral artery diameter
胆固醇对平滑肌 BK 通道蛋白的调节以及随后对脑动脉直径的控制
基本信息
- 批准号:10063416
- 负责人:
- 金额:$ 64.6万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2020
- 资助国家:美国
- 起止时间:2020-07-01 至 2024-06-30
- 项目状态:已结题
- 来源:
- 关键词:AddressAmino AcidsAnimal ModelAnteriorArteriesArteriosclerosisAtherosclerosisBindingBiological AssayBlood VesselsBrainBrain regionCalciumCaliberCell membraneCell modelCellsCerebrovascular CirculationCerebrovascular DisordersCerebrumChemosensitizationCholesterolCognitive deficitsComplementary DNAComputer ModelsConsensusCouplingDataDeformityDementiaDiseaseElectrophysiology (science)ElectroporationEndotheliumEngineeringEnsureFunctional disorderHigh PrevalenceIn VitroIncidenceInflammatoryIon ChannelKnowledgeLeadLinkLipid BilayersLipidsMass Spectrum AnalysisMediatingMethodsMicrocirculationMicrovascular DysfunctionModificationMolecularMorphologyMusMuscle ContractionMuscle TonusMuscle relaxation phaseMutateOrganPathologicPathway interactionsPatientsPharmaceutical PreparationsPharmacologyPhenotypePhysiologicalPhysiologyPotassiumPotassium ChannelPropertyProteinsRed CrossRegulationResistanceRisk FactorsRoleSarcoplasmic ReticulumScanningSecondary toSmooth MuscleSmooth Muscle MyocytesStrokeStructureTailTestingTherapeutic AgentsTissuesVasodilationVasodilator AgentsVoltage-Gated Potassium Channelanimal databasecerebral arterycholesterol controlconfocal imagingconstrictiondesignhemodynamicshuman datain vivointracranial arterylarge-conductance calcium-activated potassium channelsmiddle cerebral arterynovelparenchymal arteriolespatch clampstroke patienttraffickingvascular factorvoltage
项目摘要
Human and animal data demonstrate that cholesterol (CLR) may disrupt the smooth muscle (SM) tone of
resistance-size cerebral arteries in absence of any sign of arteriosclerosis or anatomical abnormalities, this CLR
action contributing to a vascular component of stroke. In a further step, stroke patients with dysfunction of large
cerebral arteries and high CLR will present a higher prevalence of arteriosclerosis. CLR actions on cerebral
artery SM tone have been attributed to endothelial, microcirculation, inflammatory and other circulating factors.
Departing from this current paradigm and supported by preliminary data, our central hypothesis is that CLR may
control resistance-size, cerebral artery SM tone and diameter via regulation of calcium- and voltage-gated
potassium channels of big conductance (BK) located in the cerebral artery SM plasmalemma. Here, BK function
primarily results from the coupling of channel-forming α (cbv1) and regulatory β1 subunits. The concerted action
of cbv1+β1 and eventual BK activation generates outward potassium current that opposes depolarization-
mediated calcium influx, limits SM contraction and favors artery dilation. Based on preliminary data, we predict
that under low β1 expression, CLR interaction (through direct binding and allosteric modulation) with selected
Cholesterol Recognition Amino acid Consensus motifs (CRACs) identified in the cbv1 cytosolic domain will lead
to CLR-induced reduction of BK current, SM contraction and cerebral artery constriction. In contrast, under high
β1 levels, CLR-driven increases in the plasmalemmal fraction of β1 and cbv1-β1 functional coupling will prevail,
leading to increased BK current, SM relaxation and artery dilation. We predict that brain arteries that differ in β1
expression will display a differential vulnerability to CLR, and that CLR levels in SM will condition the efficacy of
β1-dependent vasodilators. Our predictions will be tested along three specific aims (SA). SA1 (molecular level):
determine the structural bases and gating mechanisms that lead to CLR-induced hindering of BK (cbv1
homotetramer) function through CLR-cbv1 CRAC interactions. SA2 (cellular level): determine the mechanisms
that underlie CLR activation of cbv1+β1 channels. Knowledge from SA1 and SA2 will be integrated in SA3 (organ
level): determine the consequences of CLR-BK subunit interactions on the function of native BKs in cerebral
artery SM and on artery diameter under physiological conditions using in vitro and in vivo CLR delivery methods.
We combine unique expertise in computational modeling, binding assays, mass spectroscopy, differential
scanning fluorimetry, patch-clamp and lipid bilayer electrophysiology, allosteric gating analysis, electroporation
of SM cells and cerebral arteries from engineered mice with mutated cDNAs, subunit trafficking, confocal imaging
and cerebral artery diameter determinations, ensuring feasibility. We expect to challenge the paradigm that CLR
modulation of BK is secondary to nonspecific perturbation of bilayer physico-chemical properties, and to provide
milestone information on CLR control of cerebral artery function, which will be necessary to design small drugs
that adjust BK channel function to variable CLR levels and counteract CLR-associated cerebrovascular disease.
人类和动物数据表明,胆固醇(CLR)可能会扰乱血管平滑肌(SM)的张力
阻力大小的脑动脉,没有任何动脉硬化或解剖异常的迹象,这项CLR
导致中风的血管成分的动作。在进一步的步骤中,中风患者功能障碍较大
脑动脉和高CLR将呈现更高的动脉硬化患病率。CLR对大脑的作用
动脉Sm张力与内皮、微循环、炎症等循环因素有关。
从目前的范式出发,并得到初步数据的支持,我们的中心假设是,CLR可能
通过调节钙和电压门控来控制阻力-大小、脑动脉Sm音调和直径
大电导钾通道(BK)位于大脑动脉SM质膜。这里,BK函数
主要由通道形成α(CBV1)和调节β1亚基的偶联所致。协同行动
Cbv1+β1和最终的BK激活产生与去极化相反的外向钾电流-
调节钙内流,限制SM收缩,有利于动脉扩张。根据初步数据,我们预测
在β-1低表达情况下,CLR通过直接结合和变构调节与
在cbv1胞浆结构域中发现的胆固醇识别氨基酸共识基序(CRAC)将导致
对CLR引起的BK电流、SM收缩和脑动脉收缩的抑制作用。相比之下,在高
β-1水平,CLR驱动的β-1和CBV1-β-1功能偶联的血浆成分增加将占优势,
导致BK电流增加、SM松弛和动脉扩张。我们预测在β1中不同的脑动脉
表达式将显示CLR的不同漏洞,SM中的CLR水平将决定
β-1依赖的血管扩张剂。我们的预测将根据三个具体目标(SA)进行测试。SA1(分子水平):
确定导致CLR诱导的BK(Cbv1)障碍的结构基础和门控机制
同源四聚体)通过CLR-cbv1 CRAC相互作用发挥作用。SA2(细胞水平):确定机制
这是CBV1+β1通道CLR激活的基础。来自SA1和SA2的知识将被整合到SA3(器官
水平):确定CLR-BK亚基相互作用对大脑中天然BK功能的影响
动脉SM和生理条件下对动脉内径的影响采用体外和体内CLR给药方法。
我们结合了计算建模、结合分析、质谱学、差示等方面的独特专业知识
扫描荧光法、膜片钳和脂质双层电生理学、变构门控分析、电穿孔
基因工程小鼠的SM细胞和脑动脉的突变、亚单位运输、共聚焦成像
和大脑动脉直径的测定,以确保可行性。我们希望挑战CLR
BK的调制次于双层物理化学性质的非特异性扰动,并提供
CLR控制大脑动脉功能的里程碑信息,这将是设计小型药物所必需的
调节BK通道功能以适应不同的CLR水平,并对抗CLR相关性脑血管疾病。
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 }}
Anna Bukiya其他文献
Anna Bukiya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('Anna Bukiya', 18)}}的其他基金
Fetal cerebral arteries and prenatal alcohol exposure
胎儿脑动脉和产前酒精暴露
- 批准号:
10337722 - 财政年份:2022
- 资助金额:
$ 64.6万 - 项目类别:
Ionic mechanisms of toluene cerebrovascular actions
甲苯脑血管作用的离子机制
- 批准号:
10627927 - 财政年份:2022
- 资助金额:
$ 64.6万 - 项目类别:
Ionic mechanisms of toluene cerebrovascular actions
甲苯脑血管作用的离子机制
- 批准号:
10434289 - 财政年份:2022
- 资助金额:
$ 64.6万 - 项目类别:
Fetal cerebral arteries and prenatal alcohol exposure
胎儿脑动脉和产前酒精暴露
- 批准号:
10590708 - 财政年份:2022
- 资助金额:
$ 64.6万 - 项目类别:
Fetal alcohol exposure and cerebrovascular development
胎儿酒精暴露与脑血管发育
- 批准号:
10582618 - 财政年份:2021
- 资助金额:
$ 64.6万 - 项目类别:
Fetal alcohol exposure and cerebrovascular development
胎儿酒精暴露与脑血管发育
- 批准号:
10359771 - 财政年份:2021
- 资助金额:
$ 64.6万 - 项目类别:
Cholesterol regulation of smooth muscle BK channel proteins and consequent control of cerebral artery diameter
胆固醇对平滑肌 BK 通道蛋白的调节以及随后对脑动脉直径的控制
- 批准号:
10627854 - 财政年份:2020
- 资助金额:
$ 64.6万 - 项目类别:
Cholesterol regulation of smooth muscle BK channel proteins and consequent control of cerebral artery diameter
胆固醇对平滑肌 BK 通道蛋白的调节以及随后对脑动脉直径的控制
- 批准号:
10413935 - 财政年份:2020
- 资助金额:
$ 64.6万 - 项目类别:
Fetal cerebrovascular eCB system as a target of maternal alcohol consumption
胎儿脑血管eCB系统作为母体饮酒的目标
- 批准号:
8570401 - 财政年份:2014
- 资助金额:
$ 64.6万 - 项目类别:
Role of BK subunits in ethanol-cholesterol synergistic inhibition of BK channel
BK亚基在乙醇-胆固醇协同抑制BK通道中的作用
- 批准号:
8146995 - 财政年份:2010
- 资助金额:
$ 64.6万 - 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:
$ 64.6万 - 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:
$ 64.6万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
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:
$ 64.6万 - 项目类别:
Continuing Grant
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:
$ 64.6万 - 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:
$ 64.6万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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:
$ 64.6万 - 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:
$ 64.6万 - 项目类别:
Studentship
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:
$ 64.6万 - 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:
$ 64.6万 - 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:
$ 64.6万 - 项目类别: