Exploring the Therapeutic Potential of Stem Cell Biology in Gliomas

探索干细胞生物学在神经胶质瘤中的治疗潜力

• 批准号: 10262378
• 负责人: Mark Gilbert
• 金额: $ 31.18万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10262378
• 关键词: Bioinformatics; Biological; Biological Models; Biology; Brain Neoplasms; Cancer cell line; Cell Culture Techniques; Cell Line; Cells; Cellular biology; Characteristics; Ciliary Neurotrophic Factor; Clinic; Clinical; Collaborations; Computer Analysis; Cytostatics; Data; Database Management Systems; Embryo; Equilibrium; Expression Profiling; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genomics; Genotype; Glioblastoma; Glioma; Homeostasis; Human; In Vitro; Laboratories; Messenger RNA; Metabolic; MicroRNAs; Modeling; Molecular; Mus; Mutate; Normal tissue morphology; Oncogenes; Pathway interactions; Patients; Phenotype; Primary Neoplasm; Research; Resources; Serum; Signal Pathway; Signal Transduction; Study models; System; Technical Expertise; Therapeutic; Therapeutic Agents; Tretinoin; Tumor Stem Cells; Tumor Suppressor Proteins; Tumor-Derived; Ursidae Family; base; exome sequencing; experimental study; gene function; genome-wide; glioma cell line; immunoreaction; improved; in vivo; material transfer agreement; methylome; neoplastic cell; nerve stem cell; new therapeutic target; novel therapeutics; personalized medicine; pre-clinical; preclinical study; screening; self-renewal; stem cell biology; stem cell differentiation; stem cells; tool; transcriptome; transcriptome sequencing; translational study; tumor; tumor xenograft; tumorigenesis

We have performed experiments to improve our understanding of the molecular mechanisms of deregulated differentiation pathways in TSCs: The delicate balance between stem cell self-renewal and differentiation is controlled by various cell intrinsic and extrinsic factors that are critical for normal tissue homeostasis. Despite extensive phenotypic and functional similarities between GSCs and normal stem cells, the differentiation potentials of GSCs are not entirely normal. Elucidation of the differentiation pathways that are operative in both normal stem cells and GSCs will be critical for fully understanding tumorigenesis and will likely lead to novel therapeutic targets. We have also identified a set of deregulated differentiation pathways in GSCs derived from human primary GBM. Elucidation of underlying molecular mechanism will provide important clues for predicting sensitivity of differentiation therapeutic approach. Characterization of TSCs in aspect of differentiation-inducing agents further revealed the limitations of traditional glioma cell lines grown in serum. For example, retinoic acid treatment and CNTF exposure potently induce differentiation in most GBM tumor initiate cells (TICs) but not of traditional cell lines. This prompted us to question whether many of potential tumor suppressors and/or cytostatic genes previously studied in cell lines, were not recognized. Given the ever-increasing number of potential GSGs and oncogenes in glioblastoma TSCs identified from bioinformatics approaches and technical expertise of stem cell culture accumulated in the laboratories, we have set up screening systems to study the function of these genes in stem cell cultures. In addition, we have made significant progress on one of keystone projects that is to understand the genomic and molecular signaling similarities and differences between our glioma TSCs and normal neural stem cells (NSC). We have performed a very large scale study of 7 different GBM-derived TSCs and normal embryonic NSC lines under both proliferative and differentiating conditions and derived high-throughput mRNA and microRNAs profiling. Since November of 2014, when Dr Gilbert initiated the new glioma stem cell translational study project, we have created 6 more glioma-derived TSCs, introduced 3 IDH- mutated TSCs, and subsequently performed the computational analyses for characterization of the genetics and signaling pathways in these GSCs, enabling these cell lines to be used to explore the therapeutic potential of glioma stem cell biology, such as metabolic changes and immunological reactions. Additionally, we have through collaborations and Material Transfer Agreements obtained important cell lines that provide an outstanding resource for the laboratory. In total, there are now 62 human tumor lines in the NOB Laboratory. As part of this project, each of these cell lines has been extensively molecularly characterized including whole exome sequencing, analysis of the methylome and RNA sequencing. These data are being uploaded into a database system to enable analysis and selection of optimal model systems for preclinical studies for all of the research groups in the NOB.

我们已经进行了实验来提高我们对TSCs中去调控分化途径的分子机制的理解：干细胞自我更新和分化之间的微妙平衡是由各种细胞内在和外在因素控制的，这些因素对正常组织的稳态至关重要。尽管GSCs与正常干细胞之间存在广泛的表型和功能相似性，但GSCs的分化潜能并不完全正常。阐明在正常干细胞和GSCs中起作用的分化途径对于充分理解肿瘤发生至关重要，并可能导致新的治疗靶点。我们还在人类原发性GBM衍生的GSCs中发现了一组不受调控的分化途径。阐明其分子机制将为预测分化治疗方法的敏感性提供重要线索。从诱导分化剂的角度对TSCs进行表征，进一步揭示了传统胶质瘤细胞系在血清中生长的局限性。例如，维甲酸处理和CNTF暴露可有效诱导大多数GBM肿瘤起始细胞（tic）的分化，但传统细胞系却不能。这促使我们质疑以前在细胞系中研究的许多潜在的肿瘤抑制因子和/或细胞抑制基因是否未被识别。鉴于生物信息学方法和实验室积累的干细胞培养技术经验在胶质母细胞瘤TSCs中发现的潜在GSGs和致癌基因数量不断增加，我们建立了筛选系统来研究这些基因在干细胞培养中的功能。此外，我们在一个关键项目上取得了重大进展，即了解胶质瘤TSCs与正常神经干细胞（NSC）之间基因组和分子信号的异同。我们对7种不同的gbm衍生的TSCs和正常胚胎NSC系在增殖和分化条件下进行了大规模研究，并获得了高通量mRNA和microRNAs谱。自2014年11月Gilbert博士启动新的胶质瘤干细胞转化研究项目以来，我们已经创建了6个胶质瘤衍生的TSCs，引入了3个IDH突变的TSCs，并随后对这些GSCs的遗传和信号通路进行了计算分析，使这些细胞系能够用于探索胶质瘤干细胞生物学的治疗潜力，如代谢变化和免疫反应。此外，我们通过合作和材料转移协议获得了重要的细胞系，为实验室提供了出色的资源。目前，NOB实验室共有62种人类肿瘤系。作为该项目的一部分，每个细胞系都进行了广泛的分子表征，包括全外显子组测序、甲基组分析和RNA测序。这些数据正在被上传到数据库系统中，以便为NOB所有研究小组的临床前研究分析和选择最佳模型系统。