Regulation of adult human oligodendrocyte progenitors

成人少突胶质细胞祖细胞的调节

• 批准号: 7234391
• 负责人: STEVEN Alan GOLDMAN
• 金额: $ 34.21万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-12-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7234391
• 关键词: Adult; Astrocytes; Binding; Brain; Calcium; Cell Differentiation process; Cell Nucleus; Cells; Cellular biology; Characteristics; Chondroitin Sulfate Proteoglycan; Competence; Complex; Cues; Demyelinating Diseases; Elements; End Point; Environment; FGFR3 gene; Family; Fluorescence-Activated Cell Sorting; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Goals; Grant; Growth; In Vitro; Individual; Instruction; Integral Membrane Protein; Lead; Ligand Binding; Ligands; Maintenance; Mediating; Mitotic; Neurons; Numbers; Oligodendroglia; Output; Pathway interactions; Phenotype; Platelet-Derived Growth Factor; Population; Process; Production; Protein Overexpression; Protein Tyrosine Phosphatase; Protein-Serine-Threonine Kinases; Proteins; Proteolysis; RNA; RNA Interference; Receptor Activation; Recruitment Activity; Regulation; Relative (related person); Reporter; Role; Sampling; Signal Pathway; Signal Transduction; Sorting - Cell Movement; Spinal Cord; Stem cells; System; Testing; Tissues; Transcript; Transplantation; Undifferentiated; Work; autocrine; base; combinatorial; density; gliogenesis; immunoreactivity; in vivo; inhibitor/antagonist; knock-down; neurogenesis; paracrine; pleiotrophin; progenitor; promoter; receptor; relating to nervous system; research study; secretase; self-renewal; syndecan; transcription factor; white matter

DESCRIPTION (provided by applicant): In the first 4 years of this grant, we assessed the cell biology and transplantation characteristics of adult human oligodendrocyte progenitor cells. We established means for their specific isolation, using CNP2:GFP and A2B5 based fluorescence-activated cell sorting (FACS), and then assessed their lineage potential. We discovered that when removed to low density culture, the progenitors were multipotential, and gave rise to neurons as well as to gila. Thus, absent autocrine and paracrine influences on their differentiation, adult WMPCs were not restricted to oligodendrocytic fate. To assess the role of the tissue environment in regulating progenitor fate, we then investigated gene expression by adult human WMPCs. We focused on identifying those transcripts that are differentially expressed by the WMPC, relative to its local tissue environment. This process enabled us to predict ligand-receptor interactions that maintain the progenitor state, as well as those that determine whether a given cell develops into an astrocyte, oligodendrocyte, or neuron. This analysis identified a set of parallel pathways that together appear to regulate the maintenance, mobilization and differentiated fate of parenchymal progenitor cells. In particular, we noted a complex interaction of: 1) receptor tyrosine phosphatase beta/zeta signaling, as regulated by its ligands pleiotrophin and NrCAM; 2) a parallel avenue of syndecan3 regulated signaling through CASK and tbr1; 3) FGFR3-dependent signaling, likely mediated through syndecan3 proteolysis and release of CASK; and 4) a neuralin and BAMBI-suppression of BMP signals. It appears that the net output of these pathways biases adult progenitors to either self-renewal or differentiation. In this application, we propose to use a combination of protein delivery, adenoviral overexpression and lentiviral RNAi knock-down to evaluate the individual elements of these candidate systems. By this means, we intend to better define the niche for gliogenesis in the adult human white matter, and by so doing to establish both necessary and sufficient genetic targets for directing the phenotypes generated by resident progenitor cells.

描述（由申请人提供）：在该资助的前4年，我们评估了成人少突胶质细胞祖细胞的细胞生物学和移植特性。我们采用基于CNP2:GFP和A2B5的荧光激活细胞分选（FACS）建立了特异性分离方法，然后评估了它们的谱系潜力。我们发现，当移到低密度培养时，祖细胞是多电位的，并产生神经元和gila。因此，没有自分泌和旁分泌对其分化的影响，成年WMPCs并不局限于少突胶质细胞的命运。为了评估组织环境在调节祖细胞命运中的作用，我们随后研究了成人WMPCs的基因表达。我们专注于识别那些由WMPC差异表达的转录本，相对于其局部组织环境。这一过程使我们能够预测维持祖细胞状态的配体-受体相互作用，以及那些决定给定细胞是否发育成星形胶质细胞、少突胶质细胞或神经元的相互作用。该分析确定了一组平行通路，它们共同调节实质祖细胞的维持、动员和分化命运。特别是，我们注意到一个复杂的相互作用：1)受体酪氨酸磷酸酶β /zeta信号，由其配体多营养蛋白和NrCAM调节；2) syndecan3通过CASK和tbr1调控信号的平行通路；3) fgfr3依赖性信号，可能通过syndecan3蛋白水解和CASK释放介导；4)神经素和bambi抑制BMP信号。这些途径的净输出似乎使成年祖细胞倾向于自我更新或分化。在这个应用中，我们建议使用蛋白质递送、腺病毒过表达和慢病毒RNAi敲除的组合来评估这些候选系统的单个元素。通过这种方法，我们打算更好地确定成人白质中胶质细胞形成的生态位，并通过这样做来建立必要和充分的遗传靶点来指导由常驻祖细胞产生的表型。