Elucidating the function of PAX3-FOXO1 in rhabdomyosarcoma with molecular reporters and next-generation genome editing
利用分子报告基因和下一代基因组编辑阐明 PAX3-FOXO1 在横纹肌肉瘤中的功能
基本信息
- 批准号:10064134
- 负责人:
- 金额:$ 22.44万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2019
- 资助国家:美国
- 起止时间:2019-12-02 至 2022-11-30
- 项目状态:已结题
- 来源:
- 关键词:AddressBiological AssayCRISPR screenCategoriesCell LineCell SeparationCellsChemicalsChromatinChromosomal RearrangementClinical ManagementClustered Regularly Interspaced Short Palindromic RepeatsCodeCombination Drug TherapyComplementComplementary DNAComplexDNA Binding DomainDegradation PathwayDependenceDevelopmentDiseaseDropoutEnzymesEpigenetic ProcessEventExcisionExonsExposure toExpression LibraryFOXO1A geneFlow CytometryFluorescenceFollow-Up StudiesFosteringFoundationsFundingFusion Oncogene ProteinsGenesGeneticGenetic ScreeningGenetic TranscriptionGoalsGrowthHydrophobicityInterventionKnock-inKnock-outLaboratoriesLibrariesMaintenanceMalignant Childhood NeoplasmMalignant NeoplasmsMapsMeasurementMediatingMethodologyModernizationMolecularMolecular ProfilingMuscleOncogenesOncologyOncoproteinsOperative Surgical ProceduresPAX3 genePathogenesisPathway interactionsPatient-Focused OutcomesPatientsPediatric NeoplasmPharmacologyPhenocopyPhenotypePoint MutationPositioning AttributeProteinsRadiation therapyReporterReportingResearchResearch PersonnelResearch Project GrantsResolutionResourcesRhabdomyosarcomaScanningStructureSurfaceTechniquesTechnologyTestingTherapeuticTimeUndifferentiatedWithdrawaladdictionbasecofactordrug discoveryeffective therapyexperienceexperimental studygene complementationgene functiongene inductiongenetic approachgenetic resourcegenome editinggenome-wideinnovationlogarithmmulticatalytic endopeptidase complexnew therapeutic targetnext generationnovel therapeuticsparalogous genepreventresponsescreeningsoft tissuetargeted treatmenttherapeutic targettherapy developmenttranscription factortumorubiquitin-protein ligase
项目摘要
Project Abstract:
Rhabdomyosarcoma is a highly metastatic soft tissue malignancy of childhood for which new therapies are
desperately needed. The clinical management of RMS patients has been largely unchanged over the past three
decades, and is currently limited to surgical resection, radiotherapy, and combination chemotherapy. In this
context, a mechanism-based targeted therapy would have potential for a transformative impact on RMS patient
outcomes. The most common genetic event in RMS pathogenesis is a chromosomal rearrangement that
produces the PAX-fusion oncoprotein, which is a chimeric transcription factor that deregulates chromatin and
transcription to promote transformation. Our domain-focused CRISPR screens validate that RMS tumors retain
a powerful addiction to the PAX-fusion, yet strategies for direct or indirect targeting of this ‘undruggable’ protein
have yet to be successful. One obstacle in this endeavor is our incomplete understanding of the upstream and
downstream factors that support the function of the PAX-fusion, which we seek to address with the research
proposed here. Through deep molecular profiling of RMS cell lines depleted of the PAX-fusion, we have recently
developed reporters which are compatible with flow cytometry-based measurements and cell sorting. This now
allows us to perform saturating genetic screens to delineate all components of the PAX-fusion pathway in this
disease. In the first aim of this study, we will perform CRISPR exon-scanning of the endogenous PAX-fusion
locus, which is an assay we previously developed for exposing functionally important domains of cancer
maintenance genes. These experiments will define the critical subregions of the fusion oncoprotein that
deregulate transcription to sustain the block in myo-differentiation. The second aim of this proposal will leverage
our recently developed paralog domain co-targeting methodology to expose all of the critical genes, and
redundant paralogous gene pairs, that are critical for the PAX-fusion to carry out its function. The final aim of this
project will identify the critical E3 ligase that acts to restrain PAX-fusion expression in RMS cells, whose function
could be stimulated to degrade this oncoprotein. This two-year research project will employ the latest innovations
in CRISPR-based genetic screening to establish an important resource for the RMS field; a genetic foundation
for mechanism-based research of the PAX-fusion oncoprotein that will enable its pharmacological modulation
with therapeutic intent.
项目摘要:
横纹肌肉瘤是一种高度转移的儿童软组织恶性肿瘤,目前正在开发新的治疗方法
过去三年来,RMS 患者的临床管理基本上没有改变。
几十年来,目前仅限于手术切除、放疗和联合化疗。
背景下,基于机制的靶向治疗将有可能对 RMS 患者产生变革性影响
RMS 发病机制中最常见的遗传事件是染色体重排。
产生 PAX 融合癌蛋白,这是一种嵌合转录因子,可以解除染色质和
我们以领域为中心的 CRISPR 筛选验证了 RMS 肿瘤的保留。
对 PAX 融合的强烈成瘾,但直接或间接靶向这种“不可成药”蛋白质的策略
这项努力尚未取得成功的一个障碍是我们对上游和下游的不完全了解。
支持 PAX-fusion 功能的下游因素,我们寻求通过研究解决这些问题
通过对 PAX 融合蛋白耗尽的 RMS 细胞系进行深入的分子分析,我们最近发现了这一点。
开发了与基于流式细胞术的测量和细胞分选兼容的生产商。
使我们能够进行饱和遗传筛选,以描绘出 PAX 融合途径的所有组成部分
在本研究的第一个目标中,我们将对内源性 PAX 融合蛋白进行 CRISPR 外显子扫描。
位点,这是我们之前开发的一种检测方法,用于暴露癌症的重要功能域
这些实验将定义融合癌蛋白的关键亚区域。
该提案的第二个目标将利用解除转录调控以维持肌分化的阻断。
我们最近开发了旁系同源域共同靶向方法来暴露所有关键基因,以及
冗余的旁系同源基因对,对于 PAX 融合发挥其功能至关重要。
该项目将鉴定关键的 E3 连接酶,该酶可抑制 RMS 细胞中的 PAX 融合表达,其功能
这个为期两年的研究项目将采用最新的创新技术。
基于 CRISPR 的遗传筛选,为 RMS 领域建立重要资源;
用于 PAX 融合癌蛋白的基于机制的研究,这将使其药理学调节成为可能
具有治疗目的。
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)
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CHRISTOPHER VAKOC其他文献
CHRISTOPHER VAKOC的其他文献
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{{ truncateString('CHRISTOPHER VAKOC', 18)}}的其他基金
Elucidating the SCP4 pathway as a multi-catalytic signaling dependency in acute myeloid leukemia
阐明 SCP4 通路作为急性髓系白血病的多催化信号传导依赖性
- 批准号:
10753227 - 财政年份:2023
- 资助金额:
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10221648 - 财政年份:2019
- 资助金额:
$ 22.44万 - 项目类别:
Exploiting POU2F3 addiction in the tuft cell variant of small cell lung cancer
在小细胞肺癌簇细胞变体中利用 POU2F3 成瘾
- 批准号:
9980811 - 财政年份:2019
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Targeting aberrant enhancer landscapes in pancreatic cancer
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10199961 - 财政年份:2019
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Exploiting POU2F3 addiction in the tuft cell variant of small cell lung cancer
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- 批准号:
10693821 - 财政年份:2019
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Targeting aberrant enhancer landscapes in pancreatic cancer
靶向胰腺癌中的异常增强子景观
- 批准号:
9816984 - 财政年份:2019
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Exploiting POU2F3 addiction in the tuft cell variant of small cell lung cancer
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Targeting aberrant enhancer landscapes in pancreatic cancer
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