Defining and targeting the epigenetic programs involved in melanoma development

定义和瞄准参与黑色素瘤发展的表观遗传程序

基本信息

批准号：
10354080
负责人：
Keiran Smalley
金额：
$ 23.63万
依托单位：
H. LEE MOFFITT CANCER CTR & RES INST
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10354080
关键词：
Ablation Address BRAF gene Cell Death Cell Line Cell Survival Cells Cellular Stress Cutaneous Melanoma DNA Damage Detection Development Ensure Enzymes Epigenetic Process Gene Expression Profile Genetic Genetic Transcription Grant HDAC8 gene Histone Deacetylase Immune Immune Evasion Immune system Incidence Individual Lesion Link Malignant Neoplasms Mediating Melanoma Cell Modeling Mus Mutation Nature Oncogenic PTEN gene Pathogenicity Pathway interactions Pharmaceutical Preparations Prevention Prevention strategy Radiation induced damage Reporting Research Research Proposals Resistance Role Signal Transduction Skin Stress Testing The Sun Tumor-infiltrating immune cells UV Radiation Exposure UV induced UV-induced melanoma Ultraviolet Rays Work cell injury experience immunogenicity in vivo inhibitor innovation melanocyte melanoma mouse model multiple omics mutant neoantigens novel prevent programs response single-cell RNA sequencing small molecule sun damage transcriptional reprogramming tumor tumor-immune system interactions ultraviolet irradiation

项目摘要

Project summary Ultraviolet radiation (UVR) exposure has long been linked to melanoma development, with clear correlations seen between latitude and the incidence of melanoma, particularly in individuals with fair skin and a poor ability to tan. As such, sporadic cutaneous melanoma has one of the highest mutational burdens of all cancers. The high mutational burden following UVR exposure is associated with increased neoantigen load and greater immune recognition. These observations suggest that the melanocytes that initiate melanoma survive UVR - induced stress/cell damage and then avoid immune recognition. Unrelated studies in our lab focused on BRAF inhibitor resistance in melanoma identified a stress-activated epigenetic program mediated through histone deacetylase (HDAC)-8 that allowed escape from both cell death and immune recognition through transcriptional reprogramming. The objective of this proposal is to develop novel melanoma prevention strategies that target the epigenetic programs required for UVR-damaged melanocytes to avoid cell death and immune recognition. We will test the hypothesis that UVR-induced HDAC8 activation allows melanocytes to avoid cell death and immune attack, allowing cells with pathogenic mutations to survive and later develop into melanoma. This proposal is conceptually innovative in seeking to define the role of HDAC8 in the epigenetic reprogramming of melanocytes that experience UVR-induced stress and DNA damage. We will further address the role of immune evasion in the development of melanoma precursor lesions and will use unique mouse models, single cell RNA-Seq, and multi-Omics approaches to identify core pathways and regulators involved in the persistence of UVR-damaged melanocytes that constitute the precursor lesions for melanoma development. In Aim 1, we will define how HDAC8 modulates the transcriptional profile of melanocytes following UV-irradiation and will determine how this contributes to increased cell survival and reduced immunogenicity. We will then use our recently developed HDAC8 mouse model to demonstrate the role for HDAC8-induction in melanoma development and will perform single cell RNA-Seq and immune profiling to characterize how HDAC8 activation in melanocytes modulates the immune microenvironment of the skin. In Aim 2, we will determine whether inhibition of HDAC8 using genetic silencing or small molecule HDAC8 inhibitors reverses the survival and immune escape transcriptional programs that are activated in melanocytes following UVR exposure. In vivo studies will evaluate whether HDAC8 inhibitors or silencing of HDAC8 in mouse skin melanocytes are sufficient to delay the initiation of melanoma development in our BRAF mutant/PTEN-silenced melanoma model, and the role of the immune system in this response. At completion of this work we expect to have demonstrated the proof-of-principle for the development of HDAC8 inhibitors as a novel melanoma prevention strategy.

项目概要紫外线辐射 (UVR) 暴露长期以来一直与黑色素瘤的发展相关，且具有明显的相关性观察到纬度与黑色素瘤发病率之间的关系，特别是在皮肤白皙且能力较差的个体中晒黑。因此，散发性皮肤黑色素瘤是所有癌症中突变负担最高的癌症之一。这 UVR 暴露后的高突变负荷与新抗原负荷增加和更大的突变负荷相关。免疫识别。这些观察结果表明，引发黑色素瘤的黑色素细胞可以在紫外线照射下存活下来 - 诱导应激/细胞损伤，然后避免免疫识别。我们实验室的无关研究主要集中在 BRAF 黑色素瘤中的抑制剂耐药性确定了通过组蛋白介导的应激激活的表观遗传程序脱乙酰酶 (HDAC)-8 可以通过以下方式逃避细胞死亡和免疫识别：转录重编程。该提案的目的是开发新的黑色素瘤预防方法针对紫外线损伤的黑素细胞所需的表观遗传程序的策略，以避免细胞死亡和免疫识别。我们将测试以下假设：UVR 诱导的 HDAC8 激活使黑素细胞能够避免细胞死亡和免疫攻击，使带有致病突变的细胞得以生存并随后发育成黑色素瘤。该提案在概念上具有创新性，旨在定义 HDAC8 在表观遗传中的作用。经历 UVR 诱导的压力和 DNA 损伤的黑素细胞的重新编程。我们将进一步解决免疫逃避在黑色素瘤前体病变发展中的作用并将使用独特的小鼠模型、单细胞 RNA 测序和多组学方法来识别参与的核心途径和调节因子构成黑色素瘤先兆病变的紫外线损伤黑色素细胞的持续存在发展。在目标 1 中，我们将定义 HDAC8 如何调节黑素细胞的转录谱紫外线照射后，将确定这如何有助于增加细胞存活率并减少免疫原性。然后，我们将使用我们最近开发的 HDAC8 小鼠模型来演示 HDAC8 在黑色素瘤发展中的诱导，并将执行单细胞 RNA 测序和免疫分析描述黑素细胞中 HDAC8 激活如何调节皮肤的免疫微环境。在目标2，我们将确定是否使用基因沉默或小分子HDAC8来抑制HDAC8 抑制剂逆转黑色素细胞中激活的生存和免疫逃逸转录程序 UVR 照射后。体内研究将评估 HDAC8 抑制剂或 HDAC8 沉默是否在小鼠皮肤黑色素细胞足以延缓我们 BRAF 中黑色素瘤发展的开始突变/PTEN 沉默的黑色素瘤模型，以及免疫系统在该反应中的作用。完成时我们希望这项工作能够证明 HDAC8 抑制剂的开发原理新的黑色素瘤预防策略。