Unraveling the interplay between metabolism, epigenetics and stem cell fate in the hair follicle

揭示毛囊新陈代谢、表观遗传学和干细胞命运之间的相互作用

基本信息

批准号：
10266311
负责人：
Matthew Tierney
金额：
$ 2.79万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2020-12-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10266311
关键词：
Address Advanced Development Affect Aging Amino Acids Attenuated Behavior Bioenergetics Bioinformatics Biological Assay Biosensor Candidate Disease Gene Carbon Cell Compartmentation Cell Differentiation process Cell Lineage Cell Maintenance Cell physiology Cells Cellular Metabolic Process Chromatin Chromatin Structure Clustered Regularly Interspaced Short Palindromic Repeats Complex Disease Engineering Epigenetic Process Epithelial Epithelium Equilibrium Evaluation Fatty Acids Fluorescence-Activated Cell Sorting Gene Expression Genetic Transcription Glucose Glutamine Goals Growth Hair Hair follicle structure Homeostasis In Vitro Intervention Knock-out Knowledge Link Maintenance Malignant Neoplasms Mass Fragmentography Measures Mediating Metabolic Metabolic Pathway Metabolic stress Metabolic syndrome Metabolism Methods Molecular Monitor Mus Natural regeneration Nutrient Output Oxidation-Reduction Pathway interactions Pattern Periodicity Phenotype Physiological Play Population Property Recurrence Reporting Rest Role Signal Pathway Signal Transduction Skin Source Stimulus System Technology Tissues Transgenic Organisms Work adult stem cell aged attenuation cell behavior cell type chromatin immunoprecipitation chromatin remodeling comparative conditional knockout fitness histone modification improved in utero in vivo in vivo regeneration insight interest knowledge integration metabolomics new therapeutic target next generation sequencing novel preservation progenitor programs regenerative repaired response self-renewal sensor stable isotope stem cell biology stem cell fate stem cells sugar therapeutic target tissue regeneration tissue stem cells trait uptake

项目摘要

PROJECT SUMMARY Emerging evidence has uncovered a critical role for cellular metabolism as stem cells (SCs) balance self- renewal and differentiation to maintain tissue homeostasis. However, little is known mechanistically about how SCs integrate metabolic stimuli during cell fate decisions within their in vivo niches. Furthermore, several classes of metabolites are increasingly being recognized as able to influence epigenetic processes that control chromatin remodeling, including highly plastic chromatin domains that direct key transcriptional programs to maintain SC identity. Thus, achieving a better understanding of the complex crosstalk between metabolism, epigenetics and SC function is critical to identify essential signaling cascades necessary to preserve SC- mediated regenerative capacity. To achieve these goals, the murine hair follicle is an excellent system because it naturally undergoes recurrent and synchronous cycles of tissue homeostasis and regeneration. First, I will isolate and purify populations of hair follicle stem cells (HFSCs) during quiescence, activation and following their transition to short-lived, proliferative progenitors. I will then perform in vivo metabolomic profiling by directly measuring metabolite abundance using gas chromatography–mass spectrometry. Stable isotope tracing will determine metabolite turnover rates and their relative contributions from several carbon sources, including glucose sugars, fatty acids and the amino acid glutamine (Aim I). Next, I will engineer bioenergetic sensors to monitor the metabolic dynamics of HFSCs throughout the hair cycle and within in vivo physiological setting. This will be accomplished by exploiting my lab's powerful and rapid in utero lentiviral technology to deliver these sensors specifically to the skin epithelium and hair follicle (Aim II). I will apply CRISPR-mediated knockout strategies using the in utero lentiviral delivery platform to functionally assess the impact of metabolic perturbations to the metabolism and behavior of HFSCs. And finally, I will measure the impact of these interventions aimed at critical metabolic pathways on chromatin remodeling in HFSCs, clarifying the mechanistic contributions played by metabolic adaptations during the epigenetic switch governing the transition from self-renewal to lineage commitment and differentiation (Aim III). In total, this work will define the functional consequences and epigenetic mechanisms underlying metabolic reprogramming in the regenerating hair follicle, providing novel therapeutic targets that have the potential to help maintain long-term tissue homeostasis in diseases affected by SC imbalance, ranging from aging to cancer.

项目概要新的证据揭示了细胞代谢在干细胞 (SC) 平衡自我平衡过程中的关键作用。然而，对于如何维持组织稳态，人们知之甚少。 SC 在其体内生态位内的细胞命运决定过程中整合代谢刺激。越来越多的代谢物类别被认为能够影响控制表观遗传过程染色质重塑，包括高度可塑的染色质结构域，可指导关键转录程序从而更好地理解新陈代谢之间的复杂串扰，表观遗传学和 SC 功能对于识别维持 SC 所需的基本信号级联至关重要为了实现这些目标，小鼠毛囊是一个优秀的系统。因为它自然地经历组织稳态和再生的循环和同步循环。首先，我将分离和纯化处于静止、激活和激活状态的毛囊干细胞 (HFSC) 群体。在它们转变为短命的增殖祖细胞后，我将进行体内代谢组学分析。使用气相色谱-质谱法直接测量代谢物丰度。追踪将确定代谢周转率及其来自几种碳源的相对贡献，包括葡萄糖、脂肪酸和氨基酸谷氨酰胺（目标 I）接下来，我将设计生物能。传感器监测 HFSC 在整个毛发周期和体内生理内的代谢动态这将通过利用我的实验室强大而快速的子宫内慢病毒技术来实现。将这些特异性递送至皮肤上皮和毛囊（目标 II）我将应用 CRISPR 介导的传感器。使用子宫内慢病毒递送平台的敲除策略来功能评估代谢的影响最后，我将衡量这些因素的影响。针对 HFSC 染色质重塑关键代谢途径的运动，阐明了代谢适应在控制转变的表观遗传开关过程中发挥的机制贡献从自我更新到血统承诺和分化（目标三），这项工作将定义功能。再生毛发代谢重编程的后果和表观遗传机制毛囊，提供新的治疗靶点，有可能帮助维持长期组织受 SC 失衡影响的疾病（从衰老到癌症）中的稳态。