Co-opting Lef-1 and miR-26b activities to regulate dental stem cells and their progeny

共同选择 Lef-1 和 miR-26b 活性来调节牙齿干细胞及其后代

• 批准号: 10453572
• 负责人: BRAD A AMENDT
• 金额: $ 43.57万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10453572
• 关键词: Adult; Affect; Architecture; Artificial Organs; Binding; Bioinformatics; Biological Assay; Biomedical Engineering; Bromodeoxyuridine; Cell Compartmentation; Cell Differentiation process; Cell Line; Cell Maintenance; Cell Proliferation; Cell division; Cells; Cervical; ChIP-seq; Complex; Daughter; Defect; Dental; Dental Enamel; Development; Disease; Embryo; Embryonic Development; Enhancers; Epigenetic Process; Epithelial; Epithelial Cells; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Genomics; Glean; Goals; Growth; Human; In Situ Hybridization; Incisor; Injury; Knockout Mice; Knowledge; Life; Lung; Mesenchyme; MicroRNAs; Molecular; Mus; Natural regeneration; Odontoblasts; Odontogenesis; Oral; Organ; Outcome; Pathway interactions; Pattern; Phenotype; Play; Pluripotent Stem Cells; Population; Process; Production; Regulation; Regulatory Pathway; Reporter; Role; Savings; Signal Pathway; Signal Transduction; Stains; Stem Cell Factor; Testing; Tissues; Tooth regeneration; Tooth structure; Trachea; Western Blotting; Work; base; cell behavior; cell injury; cell type; craniofacial; daughter cell; epithelial stem cell; experimental study; in vivo; injured; knock-down; morphogens; mouse model; novel therapeutics; oral cavity epithelium; oral tissue; organ repair; programs; regenerative; repaired; single-cell RNA sequencing; stem; stem cell differentiation; stem cell function; stem cell niche; stem cell proliferation; stem cells; tissue regeneration; tool; transcription factor; transcriptome; transcriptome sequencing

Project Summary Tissue specific regeneration requires the function of progenitor cells that can differentiate to repair diseased or injured tissues. Affected tissues can contain stem cells and their growth and patterning are controlled by transcription factors and signaling pathways. Stem cells derived from stem cell niches contribute to the regeneration of mature tissue types in many different organs, including the trachea, lungs and teeth, amongst others. These niches are formed in developing embryos, and must be maintained throughout life by symmetric cellular divisions that produce daughter pluripotent stem cells. Another equally important behavior of the cells in a stem cell niche is the production of differentiated daughter cells by asymmetric cell division, which then take the place of damaged cells in regenerative organs, in order to allow the organ to continue to function. However, the roles of specific microRNAs (miRs) in these processes are unclear. miRs have become appreciated as playing a role in stem cell differentiation. The ability to coopt miR expression in order to reprogram and control the differentiation of naive cells into different cell types will be an important tool required to create artificial organs and repair diseased tissues, saving millions of lives and public dollars. We hypothesize that miR-26b is acting in part to regulate the stem cell niche of the lower and upper incisor by regulating Lef-1 expression, and that this mechanism may be common to many tissues that house stem cell niches. Thus, the absence of miR-26b expression prior to E14.5 allows Lef-1 expression in the oral epithelium (placode) and LaCL (stem cell niche) during development. However, after E14.5, miR-26b expression in the dental epithelium restricts Lef-1 expression and then other factors (Sox2) regulate stem cell maintenance. In order to determine how Lef-1 and miR-26b is affecting gene expression during development and cells we will pursue three aims: 1) to use our murine models to understand in vivo stem cell proliferation and differentiation by Lef-1 and miR-26b OE; and use miR-26b OE to rescue the Lef-1 OE phenotype and define the role of miR-26b OE in dental development; 2) validate molecular interactions between miR-26b, Lef- 1 and known factors involved in DESC proliferation and differentiation; 3) isolate the dental epithelial cells from WT, Lef-1 OE, miR-26b OE and rescue mice to identify new genetic pathways using RNA-Seq, ChIP-Seq. and Single cell Seq.

项目摘要 组织特定的再生需要祖细胞的功能，祖细胞可以区分以修复患病或 受伤的组织。受影响的组织可以包含干细胞，其生长和模式受到控制 转录因子和信号通路。源自干细胞壁ches的干细胞有助于 在许多不同的器官中的成熟组织类型的再生，包括气管，肺和牙齿 其他的。这些壁ni是在开发胚胎中形成的，必须通过对称来维持一生 产生女儿多能干细胞的细胞分裂。细胞的另一个同样重要的行为 在干细胞生态位中是通过非对称细胞分裂产生分化的子细胞的生产，然后 取代受损的细胞在再生器官中，以使器官继续起作用。 但是，特定microRNA（miR）在这些过程中的作用尚不清楚。 mirs已成为 赞赏在干细胞分化中发挥作用。为了使mir表达的能力 重新编程和控制幼稚细胞分为不同的细胞类型将是必需的重要工具 创建人造器官并修复患病的组织，以节省数百万的生命和公共资金。 我们假设miR-26b的作用部分是为了通过 调节LEF-1表达，并且这种机制可能是许多含有干细胞的组织 利基。因此，在E14.5之前没有miR-26b表达，允许口服LEF-1表达 发育过程中上皮（placode）和LACL（干细胞生态位）。但是，在E14.5之后，miR-26b 牙齿上皮中的表达限制LEF-1表达，然后其他因素（SOX2）调节干细胞 维护。为了确定LEF-1和miR-26b在发育过程中如何影响基因表达 细胞我们将追求三个目标：1）使用我们的鼠模型了解体内干细胞增殖 LEF-1和miR-26b OE的分化；并使用miR-26b OE营救LEF-1 OE表型和 定义miR-26b OE在牙齿发育中的作用； 2）验证miR-26b，lef-之间的分子相互作用 1和已知因素涉及DESC增殖和分化； 3）将牙齿上皮细胞从 WT，LEF-1 OE，miR-26b OE和救出小鼠，使用RNA-Seq，Chip-Seq鉴定新的遗传途径。和 单细胞seq。