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.

项目概要味觉受体细胞 (TRC) 不断地从成体干细胞/祖细胞中被替换，并且这种细胞的保真度这个过程是我们味觉相对稳定的基础。然而，许多癌症疗法会扰乱我们的味觉。味觉，我们认为这是由于味觉细胞更新的扰动造成的。与多种癌症有关，并且已经并继续开发许多药物来针对这些癌症肿瘤中的通路；这些药物总是会导致患者味觉干细胞亚群的味觉功能障碍。表达Wnt靶基因Lgr5和Hedgehog靶基因Gli1，以及Wnt和Hedgehog通路已被证明可以调节体内味觉细胞更新，因此，从长远来看，了解其功能。正如本文所提出的，Wnt 和 Hedgehog 敏感干细胞在味觉稳态中的关系将阐明关于这些祖细胞如何被导致味觉功能障碍的化疗药物破坏，并允许制定减轻味觉障碍的策略。在我们的应用中，我们建议测试以下明确的假设。小鼠周围味觉乳头中 LGR5+ 和 GLI1+ 干细胞的功能关系。假设 1：表达高水平 LGR5 的祖细胞是循环缓慢的多能干细胞，可产生快速增殖的 GLI1+/LGR5low/neg 祖细胞直接产生 TRC。假设 2：LGR5+ 细胞消融后，GLI1+ 祖细胞扩大其重建外周细胞的潜力上皮细胞并产生新的 LGR5+ 干细胞。为了测试这些想法，我们结合了体内分子遗传学、舌类器官的体外生产以及单一细胞转录组分析——我们已经熟练使用的所有方法在目标 1 中，我们测试了 LGR5 与 GLI1 祖细胞产生味觉细胞丰富类器官的能力，并进一步评估在目标 2 中，每种祖细胞类型的谱系产生依赖于 Wnt 信号传导的程度。探索 GLI1+ 祖细胞在遗传后再生味觉细胞和 LGR5+ 干细胞的能力在目标 3 中，我们将时间细粒度谱系追踪与单细胞 RNA 结合起来。测序以转录组学方式定义持续产生每种功能味道的细胞谱系细胞类型，即神经胶质样细胞和甜、苦、鲜、盐和酸 TRC。总之，我们提出的研究将。导致我们对维持我们的细胞和分子机制的理解取得重大进展味觉。