Identifying vulnerabilities and therapeutic targets in Glioma Stem Cells

识别神经胶质瘤干细胞的脆弱性和治疗靶点

• 批准号: 8948656
• 负责人: Christian Elias Badr
• 金额: $ 20.09万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2018-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8948656
• 关键词: Adult; Automobile Driving; Award; Binding; Biological Process; Biology; Blood; Brain; Brain Neoplasms; Cell Death; Cell Differentiation process; Cell Line; Cell Survival; Cell model; Cells; Central Nervous System Neoplasms; Collecting Cell; Conditioned Culture Media; Dependovirus; Development; Development Plans; Ectopic Expression; Engineering; Etiology; Faculty; Foundations; Gene Delivery; Gene Expression; Gene Expression Profile; Gene Transfer; Genes; Genomic Library; Glial Fibrillary Acidic Protein; Glioblastoma; Glioma; Goals; Health; Image; Implant; In Vitro; Libraries; Luciferases; Malignant - descriptor; Malignant Glioma; Mass Spectrum Analysis; Mediating; Mesenchymal; Mesenchymal Differentiation; Mesenchymal Stem Cells; Modeling; Molecular Target; Monitor; Mus; Neuronal Differentiation; Nude Mice; Pathway interactions; Patients; Phase; Phenocopy; Phenotype; Phosphorylation; Population; Positioning Attribute; Preclinical Testing; Property; Proteins; Proteomics; RNA Interference; Radiation; Radiation therapy; Radio; Reporter; Reporting; Research; Resistance; Role; Scientist; Serum; Serum-Free Culture Media; Signal Pathway; Simian virus 40; Site; Stem cells; Subfamily lentivirinae; System; Testing; Therapeutic; Therapeutic Effect; Time; Training; Vesicle; Work; Xenograft procedure; adeno-associated viral vector; base; cancer stem cell; career; career development; cell killing; conventional therapy; differential expression; extracellular; gene delivery system; in vivo; internal control; monolayer; mortality; novel; novel therapeutics; outcome forecast; overexpression; programs; promoter; screening; self-renewal; small hairpin RNA; stem; stem cell biology; targeted treatment; temozolomide; therapeutic target; therapy resistant; transcription factor; tumor; tumor initiation; tumorigenic

 DESCRIPTION (provided by applicant): The objective of this application is to develop the career of Dr. Christian Badr to facilitate the transition to a stable independent phase and attain his ultimate goal to become an independent scientist and establish his own research program. The career development plan created by the candidate will contribute substantially to his scientific development and further his training in proteomics, RNAi and cancer stem cells biology. The proposed opportunity in the K22 will provide the PI protected time while starting his independent Faculty position and direct him towards a successful career in developing therapeutics for brain tumors. The proposed research will also serve as the foundation for an R01 proposal prior to the end of the second year of this award. Glioma stem cells (GSC) represent a subset population within malignant gliomas, and are highly tumorigenic, intrinsically resistant to current therapies, and capable of self-renewal and differentiation into mature glioma cells (GCs). There are two major subtypes of GSCs: Proneural (PN) and Mesenchymal (MES), the latter being more aggressive and more resistant to conventional treatment. We have identified a subpopulation of GSCs, termed floating cells (FC) that could be isolated from in vitro-cultured glioma cells. These cells possess a stem-like signature with prominent mesenchymal features. They tend to be more aggressive when implanted into the brain of nude mice as compared to their parental GSCs line and have a superior resistance to radiation and therapy. Due to such properties, FC represents an ideal model to identify and test new GBM therapeutics. In Aim 1 of this proposal we will use quantitative mass spectrometry to perform a proteomic analysis on different models of PN and MES GSCs including the FC population. This work will provide a deeper characterization of the FCs and identify differentially expressed proteins/pathways in PN and MES GSCs, which could be exploited to develop new therapeutics. In Aim 2, we will use imaging-based reporters and perform an RNAi screen to identify GSCs modulators which either push GSCs to a more differentiated state, making them susceptible to conventional therapy, or simply kill these cells. Finally we will test a novel gene delivery approach based on extracellular vesicles packaged in adeno-associated virus vectors (vAAV) to deliver our therapeutic RNAi to brain tumors in mice. This work can add valuable information for understanding the biology of this elusive tumor population and plasticity between GSCs subtypes. It also has the potential to identify new vulnerabilities and therapeutic avenues for PN and MES gliomas.

 描述(申请人提供)：本申请的目标是发展克里斯蒂安·巴德尔博士的职业生涯，以促进他过渡到稳定的独立阶段，并实现他成为一名独立科学家的最终目标，并建立自己的研究计划。候选人制定的职业发展计划将大大有助于他的科学发展，并进一步促进他在蛋白质组学、RNAi和癌症干细胞生物学方面的培训。在K22的拟议机会将为PI提供保护时间，同时开始他的独立学院职位，并指导他在开发脑瘤治疗方面的成功职业生涯。拟议的研究还将作为本奖项第二年结束前R01提案的基础。胶质瘤干细胞(GSC)是恶性胶质瘤中的一个亚群，具有高度的致瘤性，对目前的治疗方法具有内在的抵抗力，能够自我更新和分化为成熟的胶质瘤细胞(GC)。有两种主要的GSCs亚型：原神经干细胞(PN)和间充质干细胞(MES)，后者更具侵袭性，对传统治疗更具抵抗力。我们已经确定了一个GSCs亚群，称为浮动细胞(FC)，可以从体外培养的胶质瘤细胞中分离出来。这些细胞具有干细胞样的特征，具有突出的间充质特征。与其亲代GSCs株相比，它们被植入裸鼠的大脑中往往更具侵略性，并且对辐射和治疗具有更好的抵抗力。由于这些特性，FC是识别和测试新的GBM疗法的理想模型。在本提案的目标1中，我们将使用定量质谱学对包括FC群体在内的不同模型的PN和MES GSCs进行蛋白质组学分析。这项工作将提供FCS的更深层次的特征，并确定PN和MES GSCs中差异表达的蛋白质/通路，这可能被利用来开发新的治疗方法。在目标2中，我们将使用基于成像的记者并进行RNAi筛选来识别GSCs调节器，这些调节器要么将GSCs推向更分化的状态，使其对传统治疗更敏感，要么简单地杀死这些细胞。最后，我们将测试一种新的基因传递方法，该方法基于包装在腺相关病毒载体(VAAV)中的细胞外小泡，将我们的治疗性RNAi传递到小鼠脑瘤中。这项工作可以为了解这一难以捉摸的肿瘤群体的生物学和GSCs亚型之间的可塑性提供有价值的信息。它还有可能识别PN和MES胶质瘤的新脆弱性和治疗途径。