A Novel Large Animal Model for Studying the Developmental Potential and Function of LGR5 Stem Cells in Vivo and in Vitro

用于研究 LGR5 干细胞体内外发育潜力和功能的新型大型动物模型

• 批准号: 10575566
• 负责人: Jorge A Piedrahita
• 金额: $ 19万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10575566
• 关键词: Ablation; Address; Alleles; Anatomy; Animal Model; Basic Science; Bleomycin; Cell Lineage; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Cochlea; Complex; Data; Development; Disease; Dose; Family suidae; Fetal Lung; Future; Gene Expression; Generations; Genes; Genetic Transcription; Guidelines; Hair follicle structure; Health; Homeostasis; Human; In Vitro; Injury; Kidney; LGR5 gene; LOX gene; Label; Liver; Lung; Maintenance; Malignant Neoplasms; Modeling; Molecular; Morphogenesis; Mus; Natural regeneration; Oral; Organ; Organoids; Pathologic; Performance; Phenotype; Physiological; Physiology; Play; Population; Preparation; Process; Regenerative capacity; Regenerative research; Regulation; Reporter; Research; Resolution; Role; Seminal; Skin; Spatial Distribution; Study models; System; Tamoxifen; Tissues; Tracer; Transgenes; Transgenic Organisms; Translating; Transplantation; Validation; adult stem cell; body system; design; dosage; fetal stem cell; gastrointestinal system; human disease; improved; in vivo; injury and repair; interest; lung injury; novel; promoter; response; somatic cell nuclear transfer; stem cells; therapy development; tissue repair; tool

Project Summary LGR5, a marker of adult and fetal stem cells in various tissues and organs, has been studied extensively for the past decades. Cells expressing LGR5 play key roles in organ/tissue development, homeostasis, regeneration, and disease, including cancer. Therefore, having the ability to identify, track, and manipulate these cells in vitro and in vivo, will allow their detailed role in normal and pathological organ function. To date, most of the research on LGR5 cells has been performed using mice, which have significant anatomical, physiological, and molecular differences from the human. Utilizing a newly developed LGR5-H2B-GFP pig line we have demonstrated its wide utility for studying the role of these unique cells and have now made seminal observations in a variety of tissues/organs. However, we have also been unable to address some key aspects due to the inability of the existing model to analyze the differentiated progeny (lineage tracking) of LGR5 cells during development, after transplantation, or during the process of injury and repair. In addition, while we have been able to inactivate both alleles of the LGR5 gene, we do not have the ability to delete the cell itself. Being able to delete the LGR5 expressing cell will increase our understanding of the importance of this unique cell (versus the LGR5 gene itself) in the phenotypes of interest. Thus, to overcome these two deficiencies, and further increase the value and impact of the original LGR5-H2B-GFP animal model, we propose to develop a highly improved LGR5 pig line that will allow labeling of the LGR5 cell, tracking of its progeny after injury, and delete it when desired. The high concordance between human and pig results to date with respect to LGR5 expression and function, not only increases the value of this line for basic research, but also for rapidly translating clinical findings to humans, further increasing the impact of this unique animal model. Successful completion of the proposed aims will generate a powerful animal model for study of translational aspects of LGR5 stem cells of the gut, lung, skin, liver, cochlea, and kidney among others.

项目摘要 LGR5是各种组织和器官中成人和胎儿干细胞的标记，已广泛研究 过去几十年。表达LGR5的细胞在器官/组织发育，体内平衡，再生中起关键作用 和疾病，包括癌症。因此，具有识别，跟踪和操纵这些细胞的能力 在体内，将允许其在正常和病理器官功能中详细的作用。迄今为止，大多数研究 在LGR5细胞上已使用小鼠进行，这些小鼠具有明显的解剖学，生理和分子 与人类的差异。利用新开发的LGR5-H2B-GFP猪线，我们已经证明了它的宽度 研究这些独特细胞的作用的实用程序，现在已经在各种 组织/器官。但是，由于无法解决 现有模型，分析发育过程中LGR5细胞的分化后代（谱系跟踪），之后 移植，或在受伤和修复过程中。另外，尽管我们能够使两者都失活 LGR5基因的等位基因，我们没有删除细胞本身的能力。能够删除LGR5 表达细胞将增加我们对这个独特细胞的重要性的理解（与LGR5基因本身相比） 在感兴趣的表型中。因此，要克服这两个缺陷，并进一步增加了价值和 原始LGR5-H2B-GFP动物模型的影响，我们建议开发高度改进的LGR5猪线 这将允许标记LGR5细胞，在受伤后跟踪其后代，并在需要时将其删除。高 关于LGR5的表达和功能，人与猪之间的一致性不仅 增加了这条线对基础研究的价值，但也用于将临床发现迅速转化为人类 进一步增加了这种独特的动物模型的影响。成功完成拟议的目标将 生成一个强大的动物模型，用于研究肠道，肺，皮肤的LGR5干细胞的翻译方面 肝脏，耳蜗和肾脏等。