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，以提高对TSC中放大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受到各种细胞内在和外在因子的控制，这对于正常组织稳态至关重要。尽管GSC和正常干细胞之间的表型和功能相似性很大，但GSC的分化势并不完全正常。在正常干细胞和GSC中阐明分化途径对于完全理解肿瘤发生至关重要，并且可能导致新的治疗靶标。我们还确定了来自人类原发性GBM的GSC中的一组放松调节的分化途径。阐明潜在的分子机制将为预测分化治疗方法的敏感性提供重要的线索。 TSC在分化诱导剂方面的表征进一步揭示了在血清中生长的传统神经胶质瘤细胞系的局限性。例如，视黄酸治疗和CNTF暴露在大多数GBM肿瘤中有效地诱导细胞（TICS），但没有传统的细胞系。这促使我们质疑是否尚未认识到先前在细胞系中研究的许多潜在抑制肿瘤和/或细胞抑制基因。鉴于从生物信息学方法和实验室中积累的干细胞培养物的技术专业知识中确定的胶质母细胞瘤TSC中的潜在GSG和癌基因不断增加，我们已经建立了筛选系统来研究这些基因在干细胞培养物中的功能。此外，我们在基石项目之一上取得了重大进展，该项目是了解我们的神经胶质瘤TSC和正常神经干细胞（NSC）之间的基因组和分子信号传导相似性和差异。我们对7种不同的GBM衍生的TSC和正常的胚胎NSC系进行了非常大规模的研究，并在增殖和分化条件下以及衍生的高通量mRNA和microRNAS分析。 Since November of 2014, when Dr Gilbert initiated the new glioma stem cell tr​​anslational 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单元线，为实验室提供了出色的资源。现在，NOB实验室中总共有62条人类肿瘤线。作为该项目的一部分，这些细胞系中的每一个都经过广泛的分子表征，包括整个外显子组测序，甲基甲基分析和RNA测序。这些数据被上传到数据库系统中，以启用用于NOB中所有研究组的临床前研究的最佳模型系统的分析和选择。