Molecular mechanisms controlling skin heterogeneity

控制皮肤异质性的分子机制

• 批准号: 10504647
• 负责人: Sarah E. Millar
• 金额: $ 57.11万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10504647
• 关键词: ATAC-seq; Acne; Address; Adipocytes; Adult; Affect; Area; Candidate Disease Gene; Cells; Characteristics; Chromatin; Chromatin Structure; DNA Methylation; DNA Modification Process; Data; Dermal; Dermis; Development; Disease; Dorsal; Ear; Embryo; Embryonic Development; Enhancers; Epidermis; Epigenetic Process; Extensor; Fetal Development; Fibroblasts; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Genetic study; Growth; Hair; Hair follicle structure; Heterogeneity; Hi-C; Histology; Human; Leg; Life; Location; Maintenance; Male Pattern Baldness; Molecular; Multiomic Data; Mus; Natural regeneration; Nature; Palmoplantar Keratosis; Pathway interactions; Pattern; Phenotype; Pigmentation physiologic function; Population; Psoriasis; Quality of life; Rare Diseases; Sampling; Scalp structure; Signal Transduction; Site; Skin; Skin graft; Sweat Glands; System; Testing; Therapeutic; Thick; Tissue-Specific Gene Expression; Variant; Vitiligo; XCL1 gene; base; bisulfite sequencing; cell type; comparative; density; differential expression; experimental study; face skin; fetal; genetic analysis; histone modification; improved; in vivo; inhibitor; loss of function; mouse genetics; mouse model; multiple omics; nerve supply; new therapeutic target; novel; postnatal; programs; regional difference; response to injury; single cell analysis; single-cell RNA sequencing; skin disorder; skin organogenesis; therapeutic target; tongue papilla; tool; transcription factor; transcriptome sequencing; wound healing

Different regions of the skin vary in their characteristics such as thickness, pigmentation, innervation, and presence, size and density of hair follicles and sweat glands, that are reflected in differential responses to injury and disease. As examples, androgenetic alopecia is limited to the scalp; acne predominates in facial skin; psoriasis is often most prominent in extensor regions; palmoplantar keratoderma is limited to palms and soles; and vitiligo can appear in symmetrical patterns. While regional characteristics of the skin are established during fetal development, positional information must be retained in the skin throughout life to allow for maintenance of regional characteristics and their re-establishment in wound healing. Positional information is known to reside in the skin dermis, but its molecular basis is poorly understood. To address this question, we propose the following Specific Aims. AIM 1: To identify candidate factors and areas of chromatin involved in establishing skin heterogeneity in embryogenesis we will analyze transcriptional profiles through single cell RNA-seq, and chromatin structure via single cell ATAC-seq, in distinct regions of developing skin to identify those that show region-specific expression or openness, respectively. AIM 2: (i) To determine which of these candidate factors and chromatin areas may also be responsible for maintaining regional skin heterogeneity in adult life, we will perform the same analyses on the corresponding areas of adult skin. (ii) We hypothesize that epigenetic mechanisms contribute to maintenance of regional skin characteristics. To test this, we will first identify patterns of DNA methylation and histone modifications that characterize developing dermis in specific skin regions by carrying out Bisulfite-seq to reveal sites of DNA methylation, and CUT&RUN for histone modifications that mark enhancers and active, repressed, or poised genes. We will then ask which of these patterns are maintained in adult dermal cells from the same regions. AIM 3: To test the functions of candidate regulators in directing and maintaining region-specific differentiation programs, we will use inducible genetic tools to delete the corresponding genes in developing or adult mouse dermis in vivo. Together, these experiments provide a comprehensive and unbiased approach to identify novel mechanisms that establish and maintain skin heterogeneity. Improved understanding of these mechanisms has potential to reveal new therapeutic targets in wound healing and in common and rare diseases that affect specific skin regions and have a major negative impact on quality of life; data obtained in this project will also inform strategies for generating specific skin types, including hair follicle- and sweat gland-bearing skin, for reparative skin grafting.

皮肤的不同区域其特征各不相同，例如厚度、色素沉着、神经分布和 毛囊和汗腺的存在、大小和密度，反映在对毛囊和汗腺的不同反应中 伤害和疾病。例如，雄激素性脱发仅限于头皮；痤疮主要集中在面部 皮肤;牛皮癣通常在伸肌区域最为突出；掌跖角化病仅限于手掌和 鞋底；白癜风可以对称出现。当皮肤的区域特征确定后 在胎儿发育期间，位置信息必须终生保留在皮肤中，以便 保持区域特征并在伤口愈合中重建它们。位置信息是 已知存在于皮肤真皮中，但对其分子基础知之甚少。为了解决这个问题，我们 提出以下具体目标。目标 1：确定参与染色质的候选因素和区域 建立胚胎发生中的皮肤异质性我们将通过单细胞分析转录谱 RNA-seq 和通过单细胞 ATAC-seq 的染色质结构，在发育中皮肤的不同区域进行识别 那些分别表现出区域特异性表达或开放性的。目标 2： (i) 确定其中哪一个 候选因素和染色质区域也可能是维持区域皮肤异质性的原因 成年生活中，我们会对成人皮肤的相应区域进行同样的分析。 (ii) 我们假设 表观遗传机制有助于维持区域皮肤特征。为了测试这一点，我们首先 识别 DNA 甲基化和组蛋白修饰的模式，这些模式是特定真皮发育的特征 通过亚硫酸氢盐测序揭示皮肤区域的 DNA 甲基化位点，并对组蛋白进行 CUT&RUN 标记增强子和活跃、抑制或平衡基因的修饰。然后我们会询问其中哪一个 来自同一区域的成体真皮细胞中维持着这种模式。目标 3：测试候选人的功能 监管者在指导和维持区域特异性分化程序时，我们将使用诱导遗传 在体内删除发育中或成年小鼠真皮中相应基因的工具。在一起，这些 实验提供了一种全面且公正的方法来确定新的机制，从而建立和 保持皮肤的异质性。加深对这些机制的理解有可能揭示新的 伤口愈合以及影响特定皮肤区域的常见和罕见疾病的治疗目标 对生活质量产生重大负面影响；该项目中获得的数据还将为以下战略提供信息： 生成特定的皮肤类型，包括带有毛囊和汗腺的皮肤，用于修复性植皮。