Molecular basis of MED12 in the pathogenesis of uterine fibroids
Molecular basis of MED12 in the pathogenesis of uterine fibroids
批准号:
10672272
负责人:
THOMAS G BOYER
金额:
$36.37万
依托单位国家:
美国
项目类别:
财政年份:
2017
资助国家:
美国
项目状态:
未结题
起止时间:
2017-08-01 至 2027-04-30
关键词:
AccountingAddressBenignBindingBiochemicalCell ReprogrammingCellsChemicalsChromosomal BreaksChromosomal InstabilityCryoelectron MicroscopyDefectDevelopmentDrug TargetingDrug usageEnhancersEnzyme KineticsEnzymesEtiologyFibroid TumorFunctional disorderGene ExpressionGeneticGenetic TranscriptionGenetic TransformationGenomic InstabilityHabitual AbortionHealth Care CostsImpairmentInduced MutationInfertilityLeiomyomaLinkMass Spectrum AnalysisMeasuresMediatorMedicalMenorrhagiaMitoticMolecularMolecular ConformationMonitorMorbidity - disease rateMuscleMutationMyometrialOperative Surgical ProceduresOutputPathogenesisPathogenicityPathway interactionsPelvic PainPharmaceutical PreparationsPhenotypePhosphotransferasesPolymerasePre-Clinical ModelPremature LaborRNA Polymerase IIRecurrenceRoleRouteSOX8 geneSomatic MutationStructural BiochemistryStructureTestingTherapeuticTherapeutic InterventionTranscription InitiationTranscriptional RegulationTreatment EfficacyUterine FibroidsWomen&aposs Healthatomic interactionsbiological adaptation to stresscell growthcell transformationclinically relevantcrosslinkdosagedriver mutationeffective therapyefficacy evaluationgenome-widein vivoinhibitorinsightmouse modelmutantneoplasticnovelnovel therapeuticspre-clinicalprogramspromoterreplication stresssegregationsmall moleculestem cell differentiationstem cell fatestem cell self renewalstem cellsstress kinasestructural biologytranscription factortumortumor growthtumor initiationtumor progressiontumorigenic
中文摘要
项目摘要/摘要
子宫肌瘤(UF)是威胁全球妇女健康的最重要的良性肿瘤。不会再有了-
UF存在长期非侵入性治疗选择,对肿瘤病因的更深入了解是开发更多的关键
有效的治疗方法。因此,这一建议是有影响的,因为它提出了一种新的病因基础
主要的UF亚型,进一步为本病的治疗干预提供了概念证明
遗传背景。UFS源于单个子宫肌层干细胞(MM SC)向肿瘤的遗传转化
种子单克隆性肿瘤生长的起始细胞(UF SC)。值得注意的是,RNA中反复出现的体细胞突变
聚合酶II(RNAPII)介体亚单位MED12约占UF的70%,但这些突变如何驱动细胞
转化和肿瘤形成尚不清楚。此前,我们发现MED12突变会破坏CycC-
介体中CDK8激酶的活性,揭示了第一例也是迄今为止唯一已知的由
这些致病突变进一步暗示了CDK8在UF发病机制中的新病因学作用。这
突破性的发现是我们最初应用程序的基础,它催生了重大进展,证明
在这一新的应用中阐明介体蛋白激酶的分子基础和治疗意义的研究
MED12突变尿路综合征发病机制中的功能障碍。在这里,我们发现MED12突变会损害CDK8
通过T环失稳的活性,导致磷蛋白质组的深刻变化和调节失调
决定多发性骨髓间充质干细胞命运的细胞生长和肌源性基因表达程序。此外,我们还证明了
MED12突变诱导的CDK8失活触发R环依赖的复制应激,提示
在这一主要的UF亚类中,基因组不稳定和新的治疗脆弱性的可能基础。
因此,我们假设MED12突变诱导的介体激酶破坏驱动肿瘤的启动
通过异常MM SC重新编程和复制应激依赖的染色体而进展
不稳定。我们进一步提出,临床上相关的ATR轴抑制剂将在
MED12突变型尿失禁的临床前模型。为了验证这一点,我们将:(1)阐明生物化学基础
MED12突变破坏了介体激活酶的活性。利用结构生物学和生物化学,我们将确定
突变体MED12对CDK8 T环稳定性和构象动力学及CycC-CDK8的影响
底物结合和催化效率;(2)阐明介体激酶破坏的分子基础
驱动UF启动。我们将把多发性骨髓间充质干细胞自我更新和分化过程中介体蛋白依赖的变化联系起来。
全基因组的增强子重新编程和改变的转录输出,并进一步询问介体蛋白激酶
破坏可以重新编程MM SCs以在体内形成UF肿瘤;(3)阐明介体的分子基础
激酶的破坏推动了UF的进展。我们将调查RNAPII启动子暂停缺陷作为基础
R-环异常增加,确定R-环诱导的复制应激是否触发有丝分裂染色体断裂,以及
评估ATR轴抑制剂在MED12突变UF临床前小鼠模型中的疗效。
英文摘要
PROJECT SUMMARY/ABSTRACT
Uterine fibroids (UFs) are the most important benign neoplastic threat to women’s health worldwide. As no long-
term non-invasive treatment option exists for UFs, deeper insight into tumor etiology is key to develop more
effective therapies. Accordingly, this proposal is impactful as it suggests a novel etiological basis for the
predominant UF subtype and further offers proof of concept for therapeutic intervention in this specific
genetic setting. UFs arise from the genetic transformation of a single myometrial stem cell (MM SC) into a tumor
initiating cell (UF SC) that seeds monoclonal tumor growth. Notably, recurrent somatic mutations in the RNA
polymerase II (RNAPII) Mediator subunit MED12 account for ~70% of UFs, but how these mutations drive cell
transformation and tumor formation is unclear. Previously, we showed that MED12 mutations disrupt CycC-
CDK8 kinase activity in Mediator, revealing the first and heretofore only known biochemical defect arising from
these pathogenic mutations and further implying a new etiological role for CDK8 in UF pathogenesis. This
breakthrough discovery was the basis for our original application which spawned major advances that justify
studies in this renewal application to clarify the molecular basis and therapeutic implications of Mediator kinase
dysfunction in the pathogenesis of MED12-mutant UFs. Herein, we show that MED12 mutations impair CDK8
activity through T-loop destabilization, leading to a profoundly altered phosphoproteome and dysregulation
of cell growth and myogenic gene expression programs that dictate MM SC fate. Further, we show that
MED12 mutation-induced CDK8 inactivation triggers R-loop-dependent replication stress, suggesting a
possible basis for genomic instability and a new therapeutic vulnerability in this dominant UF subclass.
Accordingly, we hypothesize that MED12 mutation-induced Mediator kinase disruption drives tumor initiation
and progression through aberrant MM SC reprogramming and replication stress-dependent chromosomal
instability. We further propose that clinically relevant ATR axis inhibitors will provide therapeutic benefit in a
preclinical model of MED12-mutant UFs. To test this, we will: (1) Elucidate the biochemical basis by which
MED12 mutations disrupt Mediator kinase activity. Using structural biology and biochemistry, we will determine
the impact of mutant MED12 on CDK8 T-loop stability and conformational dynamics as well as CycC-CDK8
substrate binding and catalytic efficiency; (2) Elucidate the molecular basis by which Mediator kinase disruption
drives UF initiation. We will link Mediator kinase-dependent changes in MM SC self-renewal and differentiation
with genome-wide enhancer reprogramming and altered transcriptional output and further ask if Mediator kinase
disruption can reprogram MM SCs to form UF tumors vivo; (3) Elucidate the molecular basis by which Mediator
kinase disruption drives UF progression. We will investigate RNAPII promoter pausing defects as a basis for
aberrant R-loop accrual, determine if R-loop-induced replication stress triggers mitotic chromosomal breaks, and
evaluate the efficacy of ATR axis inhibitors in a preclinical mouse model of MED12-mutant UFs.
期刊论文(2)
专著(0)
科研奖励(0)
会议论文
CDK8 and CDK19 regulate intestinal differentiation and homeostasis via the chromatin remodeling complex SWI/SNF.
CDK8和CDK19通过染色质重塑复合物SWI/SNF调节肠道分化和稳态。
DOI:
10.1172/jci158593
发表时间:
2022-10-17
期刊:
JOURNAL OF CLINICAL INVESTIGATION
影响因子:
15.9
作者:
[Dannappel, Marius, V, Zhu, Danxi, Sun, Xin, Chua, Hui Kheng, Poppelaars, Marle, Suehiro, Monica, Khadka, Subash, Sian, Terry C. C. Lim Kam, Sooraj, Dhanya, Loi, Melissa, Gao, Hugh, Croagh, Daniel, Daly, Roger J., Faridi, Pouya, Boyer, Thomas G., Firestein, Ron]
通讯作者:
Firestein, Ron
Molecular basis of MED12 in the pathogenesis of uterine fibroids
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批准号:10539362
-
项目类别:
-
资助金额:$36.37万
-
财政年份:2017
-
负责人:THOMAS G BOYER
-
依托单位:
Molecular basis of MED12 in the pathogenesis of uterine fibroids
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批准号:9237368
-
项目类别:
-
资助金额:$31.85万
-
财政年份:2017
-
负责人:THOMAS G BOYER
-
依托单位:
Molecular basis of MED12 in the pathogenesis of uterine fibroids
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批准号:9927654
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项目类别:
-
资助金额:$31.03万
-
财政年份:2017
-
负责人:THOMAS G BOYER
-
依托单位:
Mediator and epigenetic control of neuronal gene expression and differentiation
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批准号:8015297
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项目类别:
-
资助金额:$36.75万
-
财政年份:2009
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负责人:THOMAS G BOYER
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Mediator and epigenetic control of neuronal gene expression and differentiation
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资助金额:$35.28万
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财政年份:2009
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负责人:THOMAS G BOYER
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依托单位:
Mediator and epigenetic control of neuronal gene expression and differentiation
-
批准号:7590982
-
项目类别:
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资助金额:$37.07万
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财政年份:2009
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Mediator and epigenetic control of neuronal gene expression and differentiation
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资助金额:$37.13万
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依托单位:
Mediator and epigenetic control of neuronal gene expression and differentiation
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资助金额:$29.11万
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财政年份:2008
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负责人:THOMAS G BOYER
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依托单位:
Osteoblast differentiation: Interactions of Wnt, Runx2 and FGF
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批准号:6729066
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资助金额:$24.05万
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依托单位:
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批准号:7030951
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资助金额:$23.7万
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财政年份:2003
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负责人:THOMAS G BOYER
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资助金额:$24.05万
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依托单位:
海外基金