Regulation of endothelial gene expression by cerebral cavernous malformation complex
Regulation of endothelial gene expression by cerebral cavernous malformation complex
批准号:
10430055
负责人:
Miguel Alejandro Lopez-Ramirez
金额:
$18.07万
依托单位国家:
美国
项目类别:
财政年份:
2018
资助国家:
美国
项目状态:
已结题
起止时间:
2018-05-01 至 2023-04-30
关键词:
ActinsActomyosinAffectAmericanAngiogenic ProteinsBindingBinding SitesBiologicalBiologyBlood VesselsCCM1 geneCaliforniaCardiovascular systemCell Culture TechniquesCell LineCellsClinical MedicineComplementComplexCultured CellsCytoskeletonDefectDevelopmentDiseaseDown-RegulationDsRedEndothelial CellsEndotheliumEnterobacteria phage P1 Cre recombinaseEnvironmentErythrocytesEventFacultyGene ExpressionGene ProteinsGeneral PopulationGenesGeneticGenetic TranscriptionGoalsHumanIn VitroIntercellular JunctionsK-Series Research Career ProgramsKnowledgeLabelLaboratoriesLiteratureLoxP-flanked alleleMediatingMentorsMessenger RNAModelingMolecularMolecular BiologyMolecular GeneticsMolecular and Cellular BiologyMorbidity - disease rateMorphologyMusMutationMyocardiumNucleic Acid Regulatory SequencesOperative Surgical ProceduresOrganismPhenotypePositioning AttributeProcessPromoter RegionsProteinsProteomicsRecombinant ProteinsRecombinantsRegulationRepressionResearchResearch MethodologyResistanceRoleSP1 geneSignal TransductionTHBS1 geneTamoxifenTemperatureTestingThrombospondin 1Tight JunctionsTimeTrainingTransgenic MiceTransgenic OrganismsUniversitiesUp-RegulationVascular Endothelial Growth FactorsZebrafishanalogangiogenesisbrain endothelial cellcardiogenesiscell motilitycerebral cavernous malformationscollaborative environmentdesignexperienceexperimental studygain of functiongenome-widein vivoinsightmRNA Expressionmalformationmedical schoolsmortalitymultidisciplinarymutantoverexpressionpreventpromoterresponsesmall moleculestructural biologytooltranscription factortranscriptome sequencingvascular abnormality
中文摘要
项目概要/摘要
导师职业发展奖将使我有机会接受额外的培训
和必要的研究经验,以实现我的长期目标,获得一个独立的教师
在一所研究型大学的职位,并有助于心血管生物学领域。的
建议的项目是一个有价值的工具,以扩大我的知识和专业知识,在复杂的研究
方法,包括小鼠和斑马鱼遗传学/分子生物学,应用于
心血管领域加州大学圣地亚哥分校(UCSD)医学院
通过与同事互动,在多学科和高度协作的环境中进行研究,
从临床医学到结构和分子生物学。此外,金斯伯格实验室
配备了细胞,蛋白质组学和分子生物学工具,以了解细胞和分子
涉及脑海绵状畸形(CCM)的基因的蛋白产物的生物学(KRIT 1,
Krev-Interaction Trapped-1、CCM 2和PDCD 10)。为了确定主要的分子过程
参与Krit1诱导的内皮细胞表型和功能改变的损失,
以时间控制的方式在小鼠内皮中缺失Krit1。为此,转基因
携带Krit1 floxed等位基因(Krit1fl/fl)和内皮特异性他莫昔芬调节Cre的小鼠
重组酶(Pdgfb-iCreERT2)。在初步研究中,
Kruppel样因子2(KLF2),一种与Krit1对斑马鱼心脏作用有关的转录因子
血栓反应蛋白1(TSP1),一种抗血管生成蛋白,
拮抗VEGF信号传导。将进行功能增益实验,
评估TSP1的再表达是否可以逆转KRIT 1的损失影响内皮表型,
功能此外,这将通过调查TSP1基因转录是否是
在培养的细胞中由于Krit1的缺失而被抑制。由于SP1/KLF结合位点在
人和小鼠TSP1启动子区的鉴定,KLF2在TSP1下调中的作用,
对Krit1基因失活的反应。此外,斑马鱼CCM模型
将用于评估体内心血管畸形期间Tsp1丢失的影响。的
本建议中描述的研究和UCSD的环境将完成我在复杂的
研究方法,并提供有关KRIT 1基因功能的基本问题的见解
在内皮细胞和生物体水平。
英文摘要
Project Summary/Abstract
The Mentored Career Development Award will give me the opportunity to receive the additional training
and research experience necessary to achieve my long-term goal of obtaining an independent faculty
position at a research-oriented university, and contribute to the field of Cardiovascular Biology. The
proposed project is a valuable tool to extent my knowledge and expertise in complex research
methods, including mouse and zebrafish genetics/molecular biology, that are applied in the
cardiovascular field. The University of California, San Diego (UCSD) School of Medicine conducts
research in a multidisciplinary and highly collaborative environment by interacting with colleagues that
span clinical medicine to structural and molecular biology. Moreover, the Ginsberg laboratory is well
equipped with cellular, proteomic, and molecular biology tools to understand cellular and molecular
biology of the protein products of genes implicated in cerebral cavernous malformations (CCM) (KRIT1,
Krev- Interaction Trapped-1, CCM2, and PDCD10). In order to identify the major molecular processes
involved in loss of Krit1-induced altered endothelial phenotype and function, a cell culture model to
delete Krit1 in mouse endothelium in a time-controlled manner was established. To this end, transgenic
mice bearing floxed alleles of Krit1 (Krit1fl/fl) and an endothelial-specific tamoxifen-regulated Cre
recombinase (Pdgfb-iCreERT2) were used. In preliminary studies, an increase in expression of
Kruppel-like factor 2 (KLF2), a transcription factor implicated in the effects of Krit1 on zebrafish heart
development, and a decrease in expression of Thrombospondin 1(TSP1), an anti-angiogenic protein
that antagonizes VEGF signaling, was observed. Gain-of-function experiments will be performed to
assess whether re-expression of TSP1 can reverse loss of KRIT1 affects endothelial phenotype and
function. Moreover, this will be extended by investigating whether TSP1 gene transcription is
suppressed as a consequence of deletion of Krit1 in cultured cells. Since SP1/KLF binding sites in
human and mouse TSP1 promoter region were identified, the role of KLF2 in downregulation of TSP1
in response to genetic inactivation of Krit1 will be investigated. In addition, the zebrafish CCM model
will be used to assess the impact of loss of Tsp1 during cardiovascular malformations in vivo. The
studies described in this proposal and the environment at UCSD will complete my training in complex
research methods and provide insight into fundamental questions about the function of the KRIT1 gene
at the endothelial cell and organism level.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
Mechanisms of hypoxia induced exacerbation of cerebral cavernous malformations
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批准号:10520059
-
项目类别:
-
资助金额:$47.17万
-
财政年份:2021
-
负责人:Miguel Alejandro Lopez-Ramirez
-
依托单位:
Mechanisms of hypoxia induced exacerbation of cerebral cavernous malformations
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批准号:10367164
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项目类别:
-
资助金额:$48.62万
-
财政年份:2021
-
负责人:Miguel Alejandro Lopez-Ramirez
-
依托单位:
Genetic and pharmacological manipulation of HEG1-KRIT1 protein complex modulates endothelial vasoprotective functions
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批准号:10676909
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项目类别:
-
资助金额:$37.35万
-
财政年份:2020
-
负责人:Miguel Alejandro Lopez-Ramirez
-
依托单位:
Genetic and pharmacological manipulation of HEG1-KRIT1 protein complex modulates endothelial vasoprotective functions
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批准号:10229372
-
项目类别:
-
资助金额:$37.46万
-
财政年份:2020
-
负责人:Miguel Alejandro Lopez-Ramirez
-
依托单位:
Regulation of endothelial gene expression by cerebral cavernous malformation complex
-
批准号:10198991
-
项目类别:
-
资助金额:$18.07万
-
财政年份:2018
-
负责人:Miguel Alejandro Lopez-Ramirez
-
依托单位:
Regulation of endothelial gene expression by cerebral cavernous malformation complex
-
批准号:9919443
-
项目类别:
-
资助金额:$18.07万
-
财政年份:2018
-
负责人:Miguel Alejandro Lopez-Ramirez
-
依托单位:
国内基金
海外基金
由actomyosin介导的集体性细胞迁移对唇腭裂发生的影响的研究
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批准号:82360313
-
项目类别:地区科学基金项目
-
资助金额:32万元
-
批准年份:2023
-
负责人:滕藤
-
依托单位: