Stem Cells

干细胞

• 批准号: 8382154
• 负责人: HARLEY IAN KORNBLUM
• 金额: $ 14.18万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8382154
• 关键词: Accounting; Adult; Animal Model; Astrocytes; Biomedical Research; Brain Neoplasms; Cell Culture Techniques; Cell model; Cells; Childhood Brain Neoplasm; Clinical; Collection; Communities; Comparative Study; Consensus; Derivation procedure; Development; Developmental Disabilities; Developmental Process; Disease; Disease model; Elements; Embryo; Ensure; Environment; Explosion; Functional disorder; Gene Expression; Generations; Genes; Genetic; Genetic Predisposition to Disease; Genotype; Goals; Human; Human Genetics; Ice; Intellectual functioning disability; Joints; Knowledge; Link; Maintenance; Malignant Neoplasms; Mental Retardation and Developmental Disabilities Research Centers; Methodology; Mission; Molecular; Molecular Models; Mus; Mutation; Mutation Spectra; Nature; Neuroglia; Neurons; Oligodendroglia; Oncogenic; Pathologic Processes; Pathway interactions; Patients; Phenotype; Pluripotent Stem Cells; Population; Preclinical Drug Evaluation; Process; Production; Property; Quality Control; RNA; Rattus; Reproducibility; Research; Research Design; Research Personnel; Resected; Resources; Sampling; Scientist; Source; Specimen; Standardization; Stem cells; Synapses; Technology; The Cancer Genome Atlas; The Sun; Time; Tumor Stem Cells; Uncertainty; Variant; Work; base; developmental disease; disease phenotype; driving force; effective therapy; embryonic stem cell; experience; functional genomics; gene discovery; gene function; genetic manipulation; genome sequencing; genome wide association study; human disease; human embryonic stem cell; in vitro Model; induced pluripotent stem cell; interest; molecular modeling; mortality; nerve stem cell; novel; novel therapeutic intervention; stem cell differentiation; tool; tumor; tumorigenic

Rationale: Biomedical research at the present time is dominated by the paradigm of linking genes and environment to function. Recent advances in human genetics through genome-wide association analyses have greatly accelerated the disease gene discovery process. However, in this post-genome sequencing era, we are faced with the challenge of determining the cellular and organismal functions of these genes and how gene dysfunction leads or contributes to the phenotype of the disease (i.e. functional genomics). In the past, most functional genomics work was carried out through using genetic manipulations to build animal models that carry the same mutations in genes as in human diseases. However, this approach is often laborious and, more importantly, how much of the human disease can be recapitulated in those animal models remains a huge uncertainty. However, until recently, no viable alternative approaches were available. The establishment of human pluripotent embryonic stem cells (hESCs) and human induced pluripotent stem cells (h-iPSCs) is beginning to revolutionize the way to approach functional genomics, disease modeling, disease mechanistic studies, drug screening, and development of novel therapeutic interventions. Particularly, with iPSC technology, where patient-specific cells are utilized as research objects, we are finally able to utilize the genetic manipulations that nature has already generated, as well as taking into account the enormous genetic predispositions/variations that exist in the population, to develop population stratified or even personalized effective therapies. To utilize this expanding technology, IDDRC investigators have expressed the need for centralized expertise, coordination, and help with stem cell/iPSC generation, maintenance, lineage differentiation, and standardization, with the aims of building novel cellular and molecular models relevant to IDD. A strong internal consensus within the UCLA IDDRC community about the importance of these cells has become the driving force for the establishing of this new Stem Cell Core, and we have all the required expertise in place at UCLA to provide such a sen/ice. A number of IDDRC investigators are studying pediatric brain tumors with the goal of alleviating the mortality and developmental disability associated with them. In an analogous fashion to the explosion in knowledge of the genotype/phenotype relationship in genetically-based developmental disorders, similar breakthroughs are being made in the study of cancer. The Cancer Genome Atlas (TCGA) project is delineating the spectrum of mutations present in human brain tumors (http://cancergenome.nih.gov/), and there has been a large increase in the understanding of oncogenic pathways in brain tumors. However, similar to genetic developmental disorders, the study of pediatric brain tumors has been hampered by the lack of appropriate in vitro models. The recent discovery of stem cell-like cells in brain tumors (Hemmati et al., 2003), including pediatric brain tumors and the ability to propagate these highly relevant, tumorigenic cultures permits the study of molecular processes that drive these cells, the correlation of genotype and phenotype, and the development of novel potential therapies. The purpose of this new Core is to provide excellent technical support and expertise in the generation, characterization, maintenance, expansion, and lineage differentiation of human pluripotent stem cells including primarily IPSCs from patients as well as previously established hESCs (as controls and for comparative studies). In addition, due to the additional joint interest among our IDDRC investigators on brain tumors, methodologies of growing brain tumor stem cells, together with prepared tumor stem cell cultures from resected tumor specimens will also be provided by the core. In addition to the rationale outlined above, there are additional reasons for establishing a Stem Cell Core within the IDDRC. Previously, based on the consensus among scientists conducting hESC work, researchers worid-wide submitted RNA samples from their brew of cultured hESCs and a large scale gene expression array analysis was carried out. The results indicated that the most important element that accounts for variation among the different samples depended upon who had been handling the cells. Different investigators handle cells differently, which probably changed the molecular/cellular properties of the cells. Therefore, a centralized effort for stem cell production, characterization, maintenance, and expansion is very beneficial for subsequent research. This Core will provide standardization and quality control of the cells to ensure reproducibility and stability of the cell sources. In addition, based on many years of experience in studying neural stem cell (NSC), differentiation from various sources including NSCs derived from developing mouse, rat, and human embryos and adult, NSCs derived from mouse and human ESCs, as well as NSCs derived from mouse and human iPSCs, Drs. Sun and Zeng are well-situated to provide expertise concerning how to effectively differentiate human iPSCs and human ESCs first into expandable NSCs, and then subsequently into functional neurons that form synaptic network and glial cells (i.e., astrocytes and oligodendrocytes). Finally, Dr. Kornblum is among the earliest investigators studying brain tumor stem cells. He and Dr. Le Belle are very familiar with the sample (brain tumor) collection as well as the subsequent derivation of brain tumor stem cell cultures. It would be difficult for an average scientist in the IDDRC to interact with the clinicians and to have access to clinical samples in a regulated manner. Drs. Kornblum and Le Belle represent an enormous resource for the IDDRC community and will be able to handle the technical or scientific issues related to brain tumor stem cells, as well as distribution of brain tumor stem cells for many types of studies.

基本原理：目前生物医学研究的主导模式是将基因和环境与功能联系起来。人类遗传学通过全基因组关联分析的最新进展极大地加速了疾病基因的发现过程。然而，在这个后基因组测序时代，我们面临着确定这些基因的细胞和生物功能以及基因功能障碍如何导致或促成疾病表型的挑战（即功能基因组学）。在过去，大多数功能基因组学工作是通过使用基因操作来建立携带与人类疾病相同的基因突变的动物模型。然而，这种方法通常是费力的，更重要的是，在这些动物模型中可以重现多少人类疾病仍然是一个巨大的不确定性。然而，直到最近，还没有可行的替代办法。人类多能胚胎干细胞（hESC）和人类诱导多能干细胞（h-iPSC）的建立开始彻底改变功能基因组学、疾病建模、疾病机制研究、药物筛选和新型治疗干预措施开发的方式。特别是，利用iPSC技术，其中患者特异性细胞被用作研究对象，我们最终能够利用自然已经产生的遗传操作，以及考虑到群体中存在的巨大遗传倾向/变异，以开发群体分层甚至个性化的有效疗法。为了利用这种不断扩展的技术，IDDRC研究人员表示需要集中的专业知识，协调和帮助干细胞/iPSC的生成，维护，谱系分化和标准化，目的是建立与IDD相关的新型细胞和分子模型。UCLA IDDRC社区内部对这些细胞重要性的强烈共识已成为建立这种新干细胞核心的驱动力，我们在UCLA拥有提供这种服务所需的所有专业知识。 一些IDDRC研究人员正在研究儿童脑肿瘤，目的是减轻 死亡率和与之相关的发育障碍。与基于遗传的发育障碍中基因型/表型关系知识的爆炸类似，癌症研究也取得了类似的突破。癌症基因组图谱（TCGA）项目正在描绘人类脑肿瘤中存在的突变谱（http：//cancergenome.nih.gov/），并且对脑肿瘤中致癌途径的理解有了很大的增加。然而，与遗传发育障碍相似，儿科脑肿瘤的研究因缺乏合适的体外模型而受到阻碍。最近在脑肿瘤中发现了干细胞样细胞（Hemmati等人，2003），包括儿科脑肿瘤和繁殖这些高度相关的致瘤培养物的能力，允许研究驱动这些细胞的分子过程、基因型和表型的相关性以及新的潜在疗法的开发。 该新核心的目的是为人类多能干细胞的生成、表征、维持、扩增和谱系分化提供卓越的技术支持和专业知识，主要包括来自患者的IPSC以及先前建立的hESC（作为对照和用于比较研究）。此外，由于我们的IDDRC研究人员对脑肿瘤的额外共同兴趣，培养脑肿瘤干细胞的方法，以及从切除的肿瘤标本制备的肿瘤干细胞培养物也将由核心提供。 除了上述基本原理外，在IDDRC内建立干细胞核心还有其他原因。此前，基于进行hESC工作的科学家之间的共识，研究人员在世界范围内提交了来自他们培养的hESC的酿造物的RNA样品，并进行了大规模的基因表达阵列分析。结果表明，解释不同样本之间差异的最重要因素取决于谁处理了细胞。不同的研究人员处理细胞的方式不同，这可能改变了细胞的分子/细胞特性。因此，干细胞生产、表征、维持和扩增的集中努力对于后续的细胞培养是非常有益的。 research.该核心将提供细胞的标准化和质量控制，以确保细胞来源的重现性和稳定性。此外，基于多年研究神经干细胞（NSC）的经验，从各种来源分化，包括衍生自发育中的小鼠、大鼠和人胚胎和成人的NSC，衍生自小鼠和人ESC的NSC，以及衍生自小鼠和人iPSC的NSC，Sun博士和Zeng博士非常适合提供关于如何有效地将人iPSC和人ESC首先分化为可扩增的NSC的专业知识，随后转化为形成突触网络的功能性神经元和神经胶质细胞（即，星形胶质细胞和少突胶质细胞）。最后，Kornblum博士是最早研究脑肿瘤干细胞的研究者之一。他和Le贝儿博士非常熟悉样本（脑肿瘤）收集以及脑肿瘤干细胞培养物的后续衍生。IDDRC的普通科学家很难与临床医生互动，也很难以受管制的方式获得临床样本。Kornblum和Le贝儿博士为IDDRC社区提供了巨大的资源，将能够处理与脑肿瘤干细胞相关的技术或科学问题，以及脑肿瘤干细胞在许多类型研究中的分布。