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（由应用程序提供）：本申请的目的是发展Christian Badr博士的职业，以促进过渡到稳定的独立阶段，并实现他成为独立科学家并建立自己的研究计划的最终目标。候选人制定的职业发展计划将为他的科学发展做出重大贡献，并进一步培训蛋白质组学，RNAi和癌症干细胞生物学。在K22中提议的机会将提供受PI保护的时间，同时开始他的独立教师职位，并指导他成功地为开发脑肿瘤治疗的职业生涯。拟议的研究还将在此奖项的第二年结束之前成为R01提案的基础。神经胶质瘤干细胞（GSC）代表恶性神经胶质瘤中的子集群体，具有高度肿瘤性，对当前疗法具有内在抗性，并且能够自我更新和分化为成熟的神经胶质瘤细胞（GCS）。 GSC有两个主要的亚型：胸膜（PN）和间质（MES），后者更具侵略性，对常规治疗具有更大的抵抗力。我们已经确定了GSC的亚群，该子群称为浮动细胞（FC），可以从体外培养的神经胶质瘤细胞中分离出来。这些细胞具有具有突出间充质特征的茎状特征。与父母GSC系列相比，将它们植入裸鼠的大脑时，它们往往更具侵略性，并且对放射线和治疗具有较高的抗药性。由于这种特性，FC代表了识别和测试新GBM治疗的理想模型。在该提案的目标1中，我们将使用定量质谱法对包括FC种群在内的PN和MES GSC的不同模型进行蛋白质组学分析。这项工作将为FCS提供更深入的表征，并确定PN和MES GSC中不同表达的蛋白质/途径，可以探索以开发新的疗法。在AIM 2中，我们将使用基于成像的记者并执行RNAi屏幕来识别GSC调节剂，该调节剂要么将GSC推向更具差异化的状态，使其容易受到常规治疗的影响，要么简单地杀死这些细胞。最后，我们将基于包装在腺相关病毒载体（VAAV）中的细胞外蔬菜（VAAV）测试一种新型的基因递送方法，以将我们的治疗性RNAi递送至小鼠的脑肿瘤。这项工作可以增加有价值的信息，以了解这种难以捉摸的肿瘤种群的生物学以及GSC亚型之间的可塑性。它还有潜力确定PN和MES Gliomas的新漏洞和治疗途径。