Characterization of progenitor populations in adult taste epithelium

成人味觉上皮祖细胞群的表征

基本信息

批准号：
10190884
负责人：
Linda A Barlow
金额：
$ 50.93万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10190884
关键词：
Ablation Adult Affect Aging Bioinformatics Body Weight decreased Cancer Patient Cell Differentiation process Cell Lineage Cells Competence Data Desire for food Development Diphtheria Toxin Drug Targeting Dysgeusia Eating Epithelial Erinaceidae Functional disorder GLI gene Genes Genetic Grain Homeostasis Housing In Vitro Infection Injury Intestines LGR5 gene Lateral Life Light Malignant Neoplasms Mediating Mental Depression Modeling Molecular Molecular Genetics Multipotent Stem Cells Mus Natural regeneration Oranges Organoids Pathway interactions Patients Pharmaceutical Preparations Population Process Production Proliferating Publishing Quality of life Receptor Cell Signal Transduction Skin Social isolation Sodium Chloride Sum Supporting Cell Surface Taste Bud Cell Taste Buds Taste Perception Testing Tongue WNT Signaling Pathway base beta catenin cancer therapy cell regeneration cell type in vivo in vivo evaluation intestinal crypt keratinocyte patient population prevent progenitor reconstitution sensory system single-cell RNA sequencing stem stem cell function stem cell survival stem cells tongue papilla transcriptome transcriptomics tumor

项目摘要

PROJECT SUMMARY Taste receptor cells (TRCs) are continually replaced from adult stem/progenitor cells, and the fidelity of this process underlies the relative constancy of our sense of taste. However, a host of cancer therapies perturb taste and we posit this is due to perturbation of taste cell renewal. The Wnt/ß-catenin and Hedgehog pathways are implicated in scores of cancers, and many drugs have been and continue to be developed to target these pathways in tumors; these drugs invariably cause taste dysfunction for patients. Subsets of taste stem cells express the Wnt target gene Lgr5 and the Hedgehog target gene Gli1, and both Wnt and Hedgehog pathways have been shown to regulate taste cell renewal in vivo. Thus, in the long term, understanding the functional relationship of Wnt- and Hedgehog-sensitive stem cells in taste homeostasis, as proposed here, will shed light on how these progenitors are disrupted by chemotherapeutics that cause taste dysfunction, and allow development of strategies to mitigate dysgeusia. In our application, we propose to test explicit hypotheses of the functional relationship of LGR5+ and GLI1+ stem cells in the circumvallate taste papillae of mice. Hypothesis 1: Progenitors expressing high levels of LGR5 are slow cycling, multipotent stem cells that produce rapidly proliferating GLI1+/LGR5low/neg progenitors that give rise directly to TRCs. Hypothesis 2: Upon LGR5+ cell ablation, GLI1+ progenitors expand their potential to reconstitute circumvallate epithelium and give rise to new LGR5+ stem cells. To test these ideas, we combine in vivo molecular genetics, in vitro production of lingual organoids, and single cell transcriptome profiling – all approaches with which we have become skilled. In Aim 1, we test the competency of LGR5 vs GLI1 progenitors to produce taste cell-replete organoids, and further assess the degree to which lineage production by each progenitor type is dependent upon Wnt signaling. In Aim 2, we explore the capacity of GLI1+ progenitors to regenerate both taste cells and LGR5+ stem cells following genetic ablation of LGR5+ cells. In Aim 3, we combine temporally fine-grained lineage tracing with single cell RNA sequencing to transcriptomically define the cell lineages that continually produce each of the functional taste cell types, i.e., glial-like cells and sweet, bitter, umami, salt and sour TRCs. In sum, our proposed studies will lead to significant advances in our understanding of the cellular and molecular mechanisms that maintain our sense of taste.

项目摘要味觉受体细胞（TRC）连续地从成年的茎/祖细胞中取代，并且这种保真度过程是我们品味感的相对统一性的基础。但是，许多癌症疗法扰动味道，我们肯定这是由于味觉细胞更新的扰动。 Wnt/ß-catenin和刺猬途径隐含在数十种癌症中，并且许多药物已经并继续开发以针对这些肿瘤的途径；这些药物总是会引起患者的味道功能障碍。味道干细胞的子集表达Wnt靶基因LGR5和刺猬靶基因GLI1，以及Wnt和刺猬途径已显示在体内调节味觉细胞更新。从长远来看，这是理解功能的如这里提出的那样关于这些祖细胞如何受到引起味道功能障碍的化学治疗剂的破坏，并允许制定减轻肿瘤障碍的策略。在我们的应用中，我们建议检验 LGR5+和GLI1+干细胞在小鼠的味道乳头状中的功能关系。假设1：表达高水平LGR5的祖细胞是缓慢循环的多能干细胞，产生迅速增殖的GLI1+/LGR5LOW/NEG祖细胞直接引起TRC。假设2：在LGR5+细胞消融时，Gli1+祖细胞扩大了重新构造的潜力上皮并引起新的LGR5+干细胞。为了测试这些想法，我们结合了体内分子遗传学，体外产生舌形器官和单一的生产细胞转录组分析 - 我们已经熟练的所有方法。在AIM 1中，我们测试 LGR5与Gli1祖细胞产生味觉细胞恢复类器官的能力，并进一步评估每种祖细胞类型产生谱系的程度取决于Wnt信号传导。在AIM 2中，我们探索Gli1+祖细胞在遗传之后再生味细胞和LGR5+干细胞的能力 LGR5+细胞的消融。在AIM 3中，我们将暂时细粒的谱系跟踪与单细胞RNA结合在一起进行转录的测序可以定义连续产生每个功能性味道的细胞谱系细胞类型，即神经胶质细胞和甜，苦，鲜味，盐和酸的TRC。总而言之，我们提出的研究将在我们对维持我们的细胞和分子机制的理解方面取得了重大进步味觉。