Cellular and molecular regulators of melanocyte regeneration

黑素细胞再生的细胞和分子调节剂

• 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
• 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; 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

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

项目摘要： 拟议的研究试图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基因表达将是 使用单细胞RNA测序（SCRNASEQ）表征。在斑马鱼中，我们将进行纵向 在再生期间对MCSC进行采样，以发现涉及的细胞固有和外在的途径 确定MCSC命运。还将进行白癜风患者样品的screnaseq，以识别人类 MCSC并评估这些细胞中活性的信号传导。初步研究已经确定了新型候选人 MCSC生物学的途径，包括NGFR信号传导。该途径很有趣，因为NGFR是 黑色素瘤引发细胞，最近被发现是去分化的黑色素瘤细胞的定义特征 对免疫检查点抑制剂和BRAF抑制剂疗法具有抗性。我们的数据导致了假设 NGFR信号传导支持MCSCS细胞状态，并且该活性由肿瘤中的黑色素瘤细胞选择 维护和耐药性。斑马鱼中的功能研究将用于询问NGFR和其他 候选途径。怀疑在MCSC命运执行，试剂盒和巨噬细胞中很重要的途径 - 介导的，还将使用类似的方法进行探测。组合途径的操作将是 进行的以评估这些不同途径如何相互依赖以调节再生。 scrnaseq 将告知目标途径研究，作为确定新的MCSC调节信号的资源，以及 提供有关跨物种MCSC信号的保存的洞察力。实验的组合将 提供对黑素细胞干细胞的详细理解以及指导它们期间引导的对照 再生过程。 该提案的具体目的是： AIM 1：定义MCSC和其他细胞的转录变化和细胞轨迹 再生 假设：再生的特征是转录变化的广泛变化，反映了细胞状态的变化和 调节剂响应组织损伤。 Subaim 1A：绘制黑色素再生期间发生的转录和细胞变化 斑马鱼 Subaim 1B：使用白癜风和正常皮肤的Scrnaseq定义人MCSC及其后代 AIM 2：确定并在功能上分析MCSC维护中涉及的信号通路和 黑素细胞再生 假设：一个细胞信号系统网络协同调节MCSC维护，激活和 命运执行。 Subaim 2a：确定其活动在MCSC及其后代发生变化的信号通路 在再生期间 Subaim 2b：在功能上分析MCSC维护和激活中涉及的途径 Subaim 2C：研究巨噬细胞和其他中介细胞类型的作用