Dermal fibroblast/ATF3 control of skin homeostasis

真皮成纤维细胞/ATF3 控制皮肤稳态

• 批准号: 9517742
• 负责人: GIAN-PAOLO DOTTO
• 金额: $ 37.66万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-10 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9517742
• 关键词: Actinic keratosis; Address; Age; Aging; Animal Model; Atrophic; Behavior; Binding; Biochemical; Biological Assay; Biological Process; CREB1 gene; Cannabinoids; Cell Aging; Cells; Characteristics; Chronic; Clinical; Cutaneous; Cyclooxygenase Inhibitors; Dermal; Development; Diffuse; Ear; Epithelial; Family; Fibroblasts; Follow-Up Studies; Gene Deletion; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Growth; Homeostasis; Human; Image Analysis; Immune system; In Situ; Inflammation; Inflammatory; Injections; Label; Lead; Lesion; Link; Malignant Neoplasms; Mesenchymal; Methods; Mus; Neoplastic Keratinocyte; Nucleic Acid Regulatory Sequences; Play; Predisposition; Premalignant; Process; Production; Recurrence; Regulation; Regulator Genes; Risk; Role; Signal Pathway; Signal Transduction; Skin; Skin Cancer; Squamous cell carcinoma; Stromal Cells; System; Testing; Time; Tissues; Transcription Factor AP-1; Tumor Expansion; UV Radiation Exposure; Up-Regulation; Work; activating transcription factor 3; base; biological adaptation to stress; cancer risk; cell type; cellular imaging; cytokine; gene induction; in vitro Assay; in vivo; in vivo imaging; keratinocyte; knock-down; mouse model; notch protein; novel; paracrine; programs; public health relevance; screening; senescence; skin organogenesis; transcription factor; tumor; tumor growth; tumor progression; tumorigenic

DESCRIPTION (provided by applicant): Exogenous insults, like UV exposure, lead to skin changes associated with aging and increased cancer risk. We recently showed that compromised Notch/CSL signaling, in dermal fibroblasts can lead to cutaneous field cancerization, an important clinical condition, consisting of multifocal premalignant keratinocyte lesions (like actinic keratosis, AK) preceded by more widespread changes in surrounding epithelial and stromal tissues. In preliminary studies we have found that, in stroma of AK lesions in which matrix remodeling and pro-inflammatory genes are up-regulated, expression of ATF3 is decreased. ATF3 is a key stress response transcription factor with a tumor promoting function in keratinocytes. Our working hypothesis is that ATF3 plays an opposite function in dermal cells, maintaining normal skin homeostasis and suppressing keratinocyte tumor progression. 1) We will test the hypothesis that dermal fibroblast expression of ATF3 plays a suppressive function against keratinocyte tumor development. In our preliminary work we have found that mice with ubiquitous ATF3 gene deletion have increased susceptibility to dysplastic/malignant skin tumor formation and that ATF3 -/- dermal fibroblasts enhance tumorigenic behavior of weakly transformed keratinocytes. We will extend the findings by analysis of mice with dermal fibroblast- specific deletion of the ATF3 gene and extend the results to human cells, utilizing a novel in vivo imaging assay for tumor expansion that we have developed. 2) We will test the hypothesis that ATF3 is involved in control of dermal fibroblast senescence and CAF activation in antagonism with compromised Notch/CSL signaling. Stromal cells senescence results in production of diffusible factors inducing paracrine tumor growth stimulation. We have found that, in dermal fibroblasts, many senescence and CAF (S-CAF) genes are up-regulated by ATF3 silencing, while increased ATF3 suppresses their expression. We will assess whether ATF3 controls expression of these genes by direct binding in parallel with Notch/CSL and/or functioning in concert with other transcription repressing mechanisms. 3) We will test the hypothesis that methods involving up-regulation of ATF3 expression and/or function can be used for suppression of dermal fibroblast senescence and/or CAF activation. Efficacy of compounds identified by in vitro assays will be validated in vivo, in animal models and by use of human cell imaging assays.

描述（由申请人提供）：外源性损伤（例如紫外线照射）会导致与衰老相关的皮肤变化和癌症风险增加。我们最近表明，真皮成纤维细胞中的 Notch/CSL 信号传导受损可导致皮肤区域癌化，这是一种重要的临床病症，包括多灶性癌前角质形成细胞病变（如光化性角化病，AK），随后周围上皮和基质组织发生更广泛的变化。在初步研究中，我们发现，在基质重塑和促炎基因上调的 AK 病变基质中，ATF3 的表达降低。 ATF3 是一种关键的应激反应转录因子，在角质形成细胞中具有促肿瘤功能。我们的工作假设是，ATF3 在真皮细胞中发挥相反的功能，维持正常的皮肤稳态并抑制角质形成细胞肿瘤的进展。 1) 我们将检验 ATF3 的真皮成纤维细胞表达对角质形成细胞肿瘤发展发挥抑制作用的假设。在我们的初步工作中，我们发现普遍存在 ATF3 基因缺失的小鼠对发育不良/恶性皮肤肿瘤形成的敏感性增加，并且 ATF3 -/- 真皮成纤维细胞增强弱转化角质形成细胞的致瘤行为。我们将通过分析真皮成纤维细胞特异性缺失 ATF3 基因的小鼠来扩展研究结果，并将结果扩展到人类细胞，利用我们开发的一种新型体内肿瘤扩增成像测定法。 2) 我们将测试以下假设：ATF3 参与控制真皮成纤维细胞衰老和 CAF 激活，以对抗 Notch/CSL 信号转导受损。基质细胞衰老导致产生诱导旁分泌肿瘤生长刺激的扩散因子。我们发现，在真皮成纤维细胞中，许多衰老和 CAF (S-CAF) 基因因 ATF3 沉默而上调，而 ATF3 增加则抑制其表达。我们将评估 ATF3 是否通过与 Notch/CSL 平行直接结合和/或与其他转录抑制机制协同发挥作用来控制这些基因的表达。 3)我们将检验以下假设：涉及上调ATF3表达和/或功能的方法可用于抑制真皮成纤维细胞衰老和/或CAF活化。通过体外测定鉴定的化合物的功效将在体内、动物模型中以及通过使用人体细胞成像测定进行验证。