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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：
10406329
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/10406329
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 Persons 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.

项目总结味觉感受器细胞(TrCs)不断地从成体干细胞/祖细胞中被取代，而这一点的保真度过程是我们味觉相对恒定的基础。然而，许多癌症治疗方法令人不安。味觉，我们假设这是由于味觉细胞更新的扰动。Wnt/？-catenin和Hedgehog通路与数十种癌症有关，许多药物已经并将继续被开发来针对这些肿瘤中的通路；这些药物总是会导致患者的味觉障碍。味觉干细胞亚群表达Wnt靶基因Lgr5和Hedgehog靶基因Gli1，以及Wnt和Hedgehog途径已经被证明在体内调节味觉细胞的更新。因此，从长远来看，理解功能性 Wnt和Hedgehog敏感干细胞在味觉动态平衡中的关系，如本文所述，将有助于揭示关于这些祖细胞是如何被导致味觉障碍的化疗药物干扰的，并允许制定缓解地貌障碍的战略。在我们的应用程序中，我们建议测试显式假设 LGR5+和GLI1+干细胞在小鼠环状味觉乳头中的功能关系假设1：表达高水平LGR5的祖细胞是循环缓慢的多能干细胞，能产生快速增殖的GLI1+/LGR5low/neg祖细胞，可直接分化为TRCs。假设2：在LGR5+细胞消融后，GLI1+祖细胞扩大其重建环状细胞的潜力并分化为新的LGR5+干细胞。为了测试这些想法，我们结合了体内分子遗传学、体外生产舌质器官和单一的细胞转录组分析--我们已经熟练使用的所有方法。在目标1中，我们测试 LGR5与GLI1前体细胞产生味觉细胞丰富的有机化合物的能力，并进一步评估每种祖细胞类型的谱系生产在多大程度上依赖于Wnt信号。在目标2中，我们探讨GLI1+祖细胞在遗传后再生味觉细胞和LGR5+干细胞的能力 LGR5+细胞的消融。在目标3中，我们将时间上的细粒度谱系追踪与单细胞rna相结合。对持续产生每一种功能性味道的细胞谱系进行转录定义的测序细胞类型，即胶质样细胞和甜、苦、鲜味、盐和酸的TRC。总括而言，我们建议的研究将会导致我们对细胞和分子机制的理解取得重大进展，这些机制维持我们的味觉。