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（由申请人提供）：本申请的目的是发展Christian Badr博士的职业生涯，以促进其向稳定的独立阶段过渡，并实现他成为一名独立科学家并制定自己的研究发展计划的最终目标。候选人创建的项目将为他的科学发展做出重大贡献，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。K22 中拟议的机会将为 PI 提供受保护的时间，同时开始他的独立教职职位，并引导他走向成功的职业生涯。在发展中拟议的研究也将作为该奖项第二年结束之前的 R01 提案的基础，神经胶质瘤干细胞 (GSC) 代表恶性神经胶质瘤的一个子集，并且本质上具有高度致瘤性。 GSCs 有两种主要亚型：前神经细胞 (PN) 和间充质细胞 (MES)。我们已经鉴定出一种 GSC 亚群，称为浮动细胞 (FC)，可以从体外培养的神经胶质瘤细胞中分离出来，这些细胞具有类似干细胞的特征，具有显着的间质特征。与亲代 GSC 系相比，当植入裸鼠大脑时，它们往往更具攻击性，并且对放射和治疗具有优异的抵抗力，因此 FC 代表了识别和测试新 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法对包括 FC 群体在内的 PN 和 MES GSC 的不同模型进行蛋白质组分析。这项工作将提供 FC 的更深入的表征，并识别 PN 和 MES GSC 中差异表达的蛋白质/通路。 MES GSC，可用于开发新疗法在目标 2 中，我们将使用基于成像的生产者并进行 RNAi 筛选来识别 GSC 调节剂，这些调节剂可以推动 GSC 发挥作用。最后，我们将测试一种基于腺相关病毒载体 (vAAV) 中包装的细胞外囊泡的新型基因传递方法，以将我们的治疗性 RNAi 传递到脑肿瘤中。这项工作可以为了解这种难以捉摸的肿瘤群体的生物学和 GSC 亚型之间的可塑性提供有价值的信息，它还有可能确定 PN 和 MES 的新漏洞和治疗途径。神经胶质瘤。