Architecture and communication controlling the efficient generation of beta cells

控制β细胞有效生成的架构和通信

• 批准号: 8522280
• 负责人: Christopher V Wright
• 金额: $ 130.77万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-15 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8522280
• 关键词: 3-Dimensional; Apical; Architecture; Area; Basal lamina; Behavior; Beta Cell; Biological; Birth; Cell Differentiation process; Cell Polarity; Cell division; Cell surface; Cells; Cellular biology; Commit; Communication; Complement; DNA Sequence Rearrangement; Data; Data Set; Development; Duct (organ) structure; Endocrine; Epithelial; Epithelial Cells; Epithelium; Event; Extracellular Matrix; Flow Cytometry; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Generations; Genes; Genetic; In Vitro; Islet Cell; Learning; Light; Link; Location; Longevity; Maps; Mesenchymal; Molecular; Mono-S; Monoclonal Antibodies; Morphogenesis; Mus; Nature; Notch Signaling Pathway; Organogenesis; Pancreas; Population; Process; Proteins; Protocols documentation; RNA Interference; Reagent; Regulation; Regulator Genes; Resolution; Signal Pathway; Signal Transduction; Sorting - Cell Movement; Source; Staging; Stem cells; Stimulus; Structure; Testing; Time; Tissues; Transitional Cell; Work; cell type; computerized data processing; design; genetic pedigree; human embryonic stem cell; in vivo; insight; interest; intraepithelial; islet; member; migration; novel; novel marker; progenitor; programs; tool; transcriptome sequencing

DESCRIPTION (provided by applicant): We will generate translationally essential information on the process of ?-cell differentiation from progenitor cells, targeting a major outstanding issue: what stimuli and culture conditions to employ to produce mature ?-cells in vitro. Our team will focus solely on the tissue source of all ?-cell progenitors: the trunk domain epithelium of mid-pancreas organogenesis. The spatiotemporally orchestrated signaling networks and cellular division/migration processes that control ?-cell birth/maturation are very poorly defined. But, cell-autonomous and non-autonomous programs that give rise to mature b cells must work within the constraints of a defined epithelial structure, with its tightly linked morphogenetic program that produces the large numbers of budded, mature islets. We will perform a high-resolution structural and cell biological analysis of endocrine progenitor locations, cell-cell organization and signaling, and their behavior in yielding differentiating ?-cell progeny or remaining as cycling endocrine-biased cells within the epithelium. We will: (1) Define how the epithelial structure provides signaling niches that control the location, number and behavior of endocrine-biased progenitors. (2) Define how asymmetric division yields pro-endocrine precursors and endocrine-committed progeny. (3) Characterize the directional delamination of pro-endocrine cells and their progression to ?-cell or other endocrine types. (4) Localize intermediates marked by cell-instructive factors, and determine cell-autonomous and cell-non-autonomous effects. (5) Use mutational analysis and lineage tracing to detail the intraepithelial organization and signaling processes involved. (6) Perform gene expression profiling of selected cell populations in normal and perturbed situations to provide an unprecedented understanding of the gene regulatory networks involved, plus new insights into markers of critical transitional cell states. (7) Map new monoclonal antibodies from the BCBC cell-surface-marker project onto this framework, providing new tools for identifying/sorting progenitor-progeny states in differentiating hESC. (8) Test directly the functional information emerging from our studies as ?-cell formation/maturation stimuli on human ES cell (hESC) differentiating in vitro. Such a spatiotemporally resolved 'normal differentiation framework' will shed light on deficiencies in hESC differentiation protocols. In vitro hESC-derived ?-cells are immature and low in number, while partly differentiated, propancreatic cell clusters produce functional ?-cells on maturation in mice: we hypothesize that the in vitro deficit reflects an inability to build proper communication and progenitor qualities. We designed our project to integrate with and complement studies of other BCBC teams on hESC differentiation and endocrine progenitor identification. The team's pedigree in lineage analysis, cell biology and progenitor control, longevity and strong intra-BCBC interactions, led to our sharply defined common interest in epithelial endocrine progenitors. Novel tools, reagents, data sets will be shared with all BCBC members.

描述（由申请人提供）：我们将生成有关祖细胞分化过程的翻译基本信息，针对一个主要的杰出问题：在体外产生成熟的刺激和培养条件。我们的团队将仅专注于所有？ - 细胞祖细胞的组织来源：中胰腺中的躯干域上皮。控制的时空精心策划的信号网络和细胞分裂/迁移过程？ - 细胞出生/成熟的定义很差。但是，产生成熟B细胞的细胞自主和非自治程序必须在定义的上皮结构的约束中起作用，其紧密相连的形态发生程序会产生大量伙伴成熟的小岛。我们将对内分泌祖细胞位置，细胞组织和信号传导以及它们在产生区分的行为上进行高分辨率的结构和细胞生物学分析？ - 细胞后代或剩余作为上皮层内的循环内分泌偏置细胞。我们将：（1）定义上皮结构如何提供控制内分泌偏向祖细胞位置，数量和行为的信号壁ni。 （2）定义不对称的分裂如何产生前分泌前体和内分泌的后代。 （3）表征前分泌细胞的定向分层及其发展为？ - 细胞或其他内分泌类型。 （4）定位以细胞结构因子为标志的中间体，并确定细胞自主和细胞 - 非自治作用。 （5）使用突变分析和谱系跟踪来详细介绍所涉及的上皮内组织和信号传导过程。 （6）在正常和干扰情况下对选定细胞群体进行基因表达分析，以提供对所涉及的基因调节网络的前所未有的理解，以及对关键过渡细胞状态标记的新见解。 （7）将BCBC细胞表面标志物项目中的新单克隆抗体映射到该框架上，从而提供了新的工具，用于识别/分类在区分HESC中的祖细胞增强状态。 （8）直接测试我们的研究中从我们的研究中出现的功能信息，即在体外区分人类ES细胞（HESC）的细胞形成/成熟刺激。这样的时空解决的“正常分化框架”将阐明hESC分化方案中的缺陷。体外hESC衍生的？ - 细胞不成熟且数量较低，而部分分化的阳性细胞簇产生了小鼠成熟的功能？ - 我们假设体外赤字反映出无法建立适当的通信和祖细胞质量。我们设计了我们的项目，以与其他BCBC团队有关HESC分化和内分泌祖细胞鉴定的研究并补充研究。该团队在谱系分析，细胞生物学和祖细胞控制，寿命和强大的BCBC相互作用中的血统，这使我们对上皮内分泌祖细胞的共同兴趣急剧定义。新颖的工具，试剂，数据集将与所有BCBC成员共享。