Epigenetic control of skin homeostasis by the ULK3 nuclear kinase

ULK3核激酶对皮肤稳态的表观遗传控制

• 批准号: 10296312
• 负责人: GIAN-PAOLO DOTTO
• 金额: $ 47.71万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10296312
• 关键词: Acetylation; Address; Affect; Alleles; Arginine; Binding; Biochemical; Biological Assay; Cell Differentiation process; Cells; Cervical Squamous Cell Carcinoma; ChIP-seq; Chromatin; Chronic; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Co-Immunoprecipitations; Coculture Techniques; Cultured Cells; DNA-Protein Interaction; Dermal; Development; Differentiation and Growth; Disease Management; Drug Targeting; Ensure; Enzymes; Epidermis; Epigenetic Process; Erinaceidae; Fibroblasts; Future; GLI gene; GLI2 gene; Gene Deletion; Gene Expression; Gene Silencing; Gene set enrichment analysis; Genes; Genetic; Genetic Transcription; Head and Neck Squamous Cell Carcinoma; Head and neck structure; Histone H3; Histones; Homeostasis; Human; Immune; In Vitro; Individual; Lesion; Ligation; Link; Lung; Malignant Epithelial Cell; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolism; Methylation; Modeling; Morbidity - disease rate; Mus; Nuclear; Oncogenic; Organ Transplantation; Patients; Pharmacology; Phosphorylation; Phosphotransferases; Play; Prevention; Process; Promoter Regions; Protein-Arginine N-Methyltransferase; Proteins; Recurrent tumor; Regulation; Role; Signal Pathway; Signal Transduction; Site; Skin; Skin Aging; Skin Cancer; Squamous cell carcinoma; Stromal Cells; Testing; Tissues; Toxic effect; Transcriptional Regulation; Transgenes; Transgenic Mice; Transplant Recipients; UVB induced; Ultraviolet B Radiation; Work; base; carcinogenesis; cell type; clinically significant; genetic signature; in vivo; inhibitor/antagonist; interest; keratinocyte; methylome; overexpression; prevent; protein kinase inhibitor; response; self-renewal; skin cancer prevention; skin damage; skin squamous cell carcinoma; tumor progression; tumorigenic

SUMMARY Field cancerization is a major clinical condition resulting from combined keratinocyte and dermal fibroblast alterations. Identification of regulatory molecules to target alterations of both cell types is of substantial interest. We propose that the nuclear kinase ULK3 is one such a molecule, which we previously studied for its role in fibroblasts. Here we will pursue the following aims: (1) We will test the hypothesis that ULK3 impinges on key transcription regulatory networks determining keratinocyte self-renewal and oncogenic potential. ULK3 is up- regulated in keratinocyte-derived skin and head/neck Squamous Cell Carcinomas (SCCs) and silencing of the gene suppresses proliferative and tumorigenic potential of both human keratinocytes (HKCs) and SCC cells. We will explore the interplay between ULK3 and the p63 and GLI transcription networks in growth/differentiation of these cells and their oncogenicity, in orthotopic models of skin cancer expansion. 2) We will test the hypothesis that ULK3 functions at the intersection between the arginine methylome and chromatin organization. Arginine methylation is a little investigated mode of chromatin regulation in SCCs development. We have found that ULK3 associates with the arginine methylases PRMT1 and PRMT5 (PRMTs) and that ULK3 loss in SCC cells results in down-modulation of di-methylated forms of Histone 4 (H4R3), well established products of PRMTs activity. We will test whether ULK3 controls PRMT1 and 5 activity and/or links these enzymes to key substrates such as p63, GLI2 and histones (specifically H4). We will further determine whether ULK3 control of gene transcription in HKCs and SCC involves a broader impact on chromatin organization through convergent PRMT1- and 5- dependent mechanisms. 3) We will test the hypothesis that ULK3 is a target of translational relevance for skin field cancerization. We will undertake a dual genetic and pharmacological approach. Genetically, we have developed mice with Ulk3flox alleles and K14- CreER transgene for keratinocyte-specific Ulk3 deletion. We will examine the impact of Ulk3 deletion on normal skin homeostasis and, after transfer into an albino hairless background, assess the consequences for UVB-induced skin field cancerization. Pharmacologically, we found that ULK3 inhibitory compounds suppress proliferative potential of SCC cells as well as cancer/stromal cell expansion. We will extend our studies with ULK3 and PRMT1/5 inhibitors on cultured cells and further test the most promising compounds in an in vivo orthotopic model of skin cancer/stromal cells expansion.

概括 局部癌化是由角质形成细胞和真皮成纤维细胞联合引起的主要临床病症 变更。鉴定针对两种细胞类型的靶向改变的调节分子具有重大意义。 我们认为核激酶 ULK3 就是这样一种分子，我们之前研究过它在 成纤维细胞。在这里，我们将追求以下目标：（1）我们将检验 ULK3 影响关键的假设 转录调控网络决定角质形成细胞的自我更新和致癌潜力。 ULK3 已上线 - 在角质形成细胞衍生的皮肤和头/颈鳞状细胞癌 (SCC) 中受到调节，并沉默 该基因抑制人角质形成细胞 (HKC) 和 SCC 细胞的增殖和致瘤潜力。我们 将探索 ULK3 与 p63 和 GLI 转录网络在生长/分化中的相互作用 在皮肤癌扩张的原位模型中，这些细胞及其致癌性。 2）我们将检验假设 ULK3 在精氨酸甲基化组和染色质组织之间的交叉点发挥作用。精氨酸 甲基化是 SCC 发育过程中染色质调控的研究较少的模式。我们发现ULK3 与精氨酸甲基化酶 PRMT1 和 PRMT5 (PRMT) 相关，并且导致鳞状细胞癌细胞中 ULK3 缺失 下调组蛋白 4 (H4R3) 的二甲基化形式，组蛋白 4 (H4R3) 是 PRMT 活性的成熟产物。 我们将测试 ULK3 是否控制 PRMT1 和 5 活性和/或将这些酶与关键底物联系起来，例如 p63、GLI2 和组蛋白（特别是 H4）。我们将进一步确定ULK3是否控制基因转录 在 HKC 和 SCC 中，通过聚合 PRMT1- 和 5- 对染色质组织产生更广泛的影响 依赖机制。 3) 我们将检验 ULK3 是皮肤翻译相关性靶标的假设 野癌化。我们将采取双重遗传和药理学方法。从基因上来说，我们有 开发了带有 Ulk3flox 等位基因和 K14-CreER 转基因的小鼠，用于角质形成细胞特异性 Ulk3 缺失。我们将 检查 Ulk3 缺失对正常皮肤稳态的影响，并在转移到无毛白化病患者后 背景，评估 UVB 诱导的皮肤区域癌化的后果。从药理学上我们发现 ULK3 抑制化合物可抑制 SCC 细胞以及癌症/基质细胞的增殖潜力 扩张。我们将在培养细胞上扩展 ULK3 和 PRMT1/5 抑制剂的研究，并进一步测试 在皮肤癌/基质细胞扩张的体内原位模型中最有前途的化合物。