Cellular and molecular regulators of melanocyte regeneration

黑素细胞再生的细胞和分子调节剂

基本信息

批准号：
10659536
负责人：
Craig Joseph Ceol
金额：
$ 35.55万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10659536
关键词：
Ablation Adopted Affect Autoimmune Diseases Autologous Transplantation BRAF gene Biological Assay Biological Models Bulla Cell Lineage Cell Maintenance Cells Data Disease Dissection Drug resistance Gene Expression Genes Genetic Genetic Transcription Homeostasis Human Hypopigmentation Immersion Immune checkpoint inhibitor Individual Knowledge Lead Lesion Macrophage Maintenance Maps Measures Mediating Melanoma Cell Methodology Molecular NGFR gene Natural regeneration Pathway interactions Patients Pharmacotherapy Phototherapy Population Procedures Process Recovery Resistance Resources Role Sampling Signal Pathway Signal Transduction Skin Skin Pigmentation Stigmatization System Techniques Time Tissues Ultraviolet Rays Vitiligo Zebrafish cell type combinatorial experimental study inhibitor therapy insight melanocyte melanoma novel patient subsets psychologic regenerative cell repaired response single-cell RNA sequencing social stem cell biology stem cell fate stem cell genes stem cell self renewal stem cells tissue injury tissue regeneration transcriptomics tumor

项目摘要

PROJECT SUMMARY: The proposed studies seek to isolate and characterize melanocyte stem cells (McSCs) as well as identify the signals and pathways that govern McSC-mediated melanocyte regeneration. McSC gene expression will be characterized using single-cell RNA sequencing (scRNAseq). In zebrafish we will perform longitudinal sampling of McSCs during regeneration to uncover pathways, both cell intrinsic and extrinsic, involved in determining McSC fates. scRNAseq of samples from vitiligo patients will also be performed to identify human McSCs and assess signaling active in these cells. Preliminary studies have identified novel candidate pathways in McSC biology, including NGFR signaling. This pathway is intriguing because NGFR is a marker of melanoma initiating cells and was recently found as a defining feature of dedifferentiated melanoma cells that are resistant to both immune checkpoint inhibitor and BRAF inhibitor therapies. Our data lead to the hypothesis that NGFR signaling supports the McSCs cell state and this activity is co-opted by melanoma cells in tumor maintenance and drug resistance. Functional studies in zebrafish will be used to interrogate NGFR and other candidate pathways. Pathways suspected to be important in McSC fate execution, KIT and macrophage- mediated, will be also probed using similar methodologies. Combinatorial pathway manipulations will be performed to assess how these different pathways depend on each other to regulate regeneration. scRNAseq will inform the targeted pathway studies, serve as a resource to identify new McSC-regulatory signals, and provide insight into the conservation of McSC signaling across species. This combination of experiments will provide a detailed understanding of melanocyte stem cells and the controls that guide them during the regeneration process. The specific aims of this proposal are: Aim 1: Define transcriptional changes and cellular trajectories of McSCs and other cells during regeneration Hypothesis: Regeneration is characterized by broad transcriptional changes that reflect cell state changes and engagement of regulators in response to tissue injury. Subaim 1A: Map the transcriptional and cellular changes that occur during melanocyte regeneration in zebrafish Subaim 1B: Define human McSCs and their descendants using scRNAseq of vitiligo and normal skin Aim 2: Determine and functionally analyze signaling pathways involved in McSC maintenance and melanocyte regeneration Hypothesis: A network of cellular signaling systems coordinately regulates McSC maintenance, activation and fate execution. Subaim 2A: Identify signaling pathways whose activities change in McSCs and their descendants during regeneration Subaim 2B: Functionally analyze pathways involved in McSC maintenance and activation Subaim 2C: Investigate the roles of macrophages and other intermediary cell types in regeneration

项目概要：拟议的研究旨在分离和表征黑素细胞干细胞 (McSC)，并确定控制 McSC 介导的黑素细胞再生的信号和途径。 McSC 基因表达将使用单细胞 RNA 测序 (scRNAseq) 进行表征。在斑马鱼中，我们将进行纵向在再生过程中对 McSC 进行采样，以揭示参与细胞内在和外在的途径决定MCSC的命运。还将对白癜风患者的样本进行 scRNAseq 鉴定人类 McSC 并评估这些细胞中的信号传导活性。初步研究已经确定了新的候选者 McSC 生物学中的通路，包括 NGFR 信号传导。这条通路很有趣，因为 NGFR 是黑色素瘤起始细胞，最近被发现是去分化黑色素瘤细胞的一个定义特征，对免疫检查点抑制剂和 BRAF 抑制剂疗法均具有耐药性。我们的数据得出了假设 NGFR 信号传导支持 McSC 细胞状态，并且这种活性被肿瘤中的黑色素瘤细胞所选择维持和耐药性。斑马鱼的功能研究将用于询问 NGFR 和其他候选途径。疑似在 McSC 命运执行、KIT 和巨噬细胞中重要的途径介导的，也将使用类似的方法进行探讨。组合途径操作将是进行评估这些不同的途径如何相互依赖来调节再生。单链RNA测序将为目标途径研究提供信息，作为识别新的 McSC 监管信号的资源，以及提供对跨物种 McSC 信号保护的深入了解。这种实验组合将提供对黑素细胞干细胞以及在黑素细胞干细胞生长过程中指导它们的控制的详细了解再生过程。该提案的具体目标是：目标 1：定义 McSC 和其他细胞在转录过程中的转录变化和细胞轨迹再生假设：再生的特点是广泛的转录变化，反映细胞状态的变化和监管机构参与应对组织损伤。 Subaim 1A：绘制黑色素细胞再生过程中发生的转录和细胞变化图斑马鱼 Subaim 1B：使用白癜风和正常皮肤的 scRNAseq 定义人类 McSC 及其后代目标 2：确定和功能分析参与 McSC 维护和治疗的信号通路黑色素细胞再生假设：细胞信号系统网络协调调节 McSC 的维持、激活和命运执行。 Subaim 2A：识别在 McSC 及其后代中活性发生变化的信号通路再生期间 Subaim 2B：功能分析涉及 McSC 维持和激活的途径 Subaim 2C：研究巨噬细胞和其他中间细胞类型在再生中的作用