Defining the epigenetic landscape in human prostate cancer

定义人类前列腺癌的表观遗传景观

• 批准号: 9438502
• 负责人: MYLES A BROWN
• 金额: $ 60.08万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9438502
• 关键词: Affect; Androgen Receptor; Androgen Suppression; Androgens; Atlases; Automobile Driving; Binding; Binding Sites; Biological; Biopsy; Cancer Etiology; Castration; Cell Line; Cells; Cessation of life; Clinical; Complex; DNA; DNA Binding; Dana-Farber Cancer Institute; Data; Data Quality; Data Set; Development; Disease; Disease Progression; Disease Resistance; Drug resistance; Epigenetic Process; Epithelial Cells; Epithelium; Experimental Models; Freezing; Gene Targeting; Genes; Genetic; Goals; High-Throughput Nucleotide Sequencing; Human; Investigation; LNCaP; Link; Localized Disease; Lymph Node Tissue; Malignant Neoplasms; Malignant neoplasm of prostate; Maps; Measures; Metastatic Neoplasm to Lymph Nodes; Metastatic to; Modeling; Neoplasm Metastasis; Nuclear; Pathway interactions; Play; Process; Prostate; Protocols documentation; Reproducibility; Research Infrastructure; Resistance; Sampling; Signal Transduction; Site; Specimen; Techniques; Therapeutic Intervention; Time; Tissue Sample; Tissues; Transcriptional Regulation; Tumor Tissue; United States; VCaP; Work; Xenograft Model; androgen sensitive; biomarker development; cancer drug resistance; chromatin immunoprecipitation; clinically relevant; disease natural history; epigenome; experimental study; genome editing; genome-wide; human tissue; innovation; insight; knock-down; men; new therapeutic target; novel; overexpression; programs; prostate cancer cell line; prostate cancer progression; prostate carcinogenesis; public health relevance; small hairpin RNA; therapy development; transcription factor; transcriptome sequencing; tumor; tumorigenesis

 DESCRIPTION (provided by applicant): The androgen receptor (AR) is central to prostate cancer development, progression, and drug resistance. The AR is a nuclear transcription factor (TF) that binds to DNA and regulates gene activity. The set of genome-wide AR-DNA binding sites is termed the AR cistrome. A complex interplay between AR and its co-regulators determines the genes targeted for transcriptional regulation. Using newly developed techniques for AR chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) in human prostate specimens, we unambiguously show that the AR program is highly dynamic and is determined, at least in part, by the presence of available co-regulators. In particular, the pioneer TF FOXA1 and the prostate lineage- specific TF HOXB13 appear to co-localize at tumor tissue-specific AR sites. The overall objectives of the present proposal are to characterize the mechanisms underlying AR reprogramming during tumorigenesis and - for the first time - to characterize the AR program in human tissue during progression from localized prostate cancer to metastatic, drug-resistant disease. In the first aim, ChIP-seq will be performed in cell line models for AR-expressing normal prostate epithelium (LHSAR) and prostate cancer (LNCaP and VCaP). Changes in the cistromes of AR, FOXA1 and HOXB13 will be measured as each TF is knocked down via shRNA and genome editing or overexpressed via lentiviral transduction. RNA-seq will also be performed for each cell line condition to determine the genes affected by epigenetic reprogramming. In the second and third aims, the epigenetic landscape of the AR will be systematically charted via ChIP-seq in human specimens: localized tissue, untreated metastatic tissue, castration-resistant metastases and enzalutamide-resistant disease. Completion of the novel experiments outlined in this proposal will provide an unparalleled look at how master transcription factors drive prostate cancer progression. Specifically, we will discover: (i) how the AR is reprogrammed during prostate cancer development and progression, and which co-regulators are facilitating this process, (ii) how the AR is reprogrammed during the acquisition of resistance to enzalutamide and, if so, which co-regulators are facilitating this process, and (iii) other non-AR master regulators that are driving prostate cancer progression. The proposed experiments will also enable us to identify the target genes that are affected by AR reprogramming. The project will result in an atlas of the epigenetic landscape as the disease progresses to the castration-resistant metastatic state, which is uniformly fatal. The genetic loci and target genes comprising this dataset will stimulate new targets for therapeutic intervention.

 描述（由适用提供）：雄激素受体（AR）对于前列腺癌发展，进展和耐药性至关重要。 AR是与DNA结合并调节基因活性的核转录因子（TF）。全基因组AR-DNA结合位点的集合称为AR cistrome。 AR及其共同调节器之间的复杂相互作用决定了针对转录调控的基因。使用新开发的AR染色质免疫沉淀技术，然后在人前列腺样本中进行高通量测序（CHIP-SEQ），我们明确地表明，AR程序具有高度动态性，并且至少部分地确定了可用的共同监管因子。特别是 先锋TF FOXA1和前列腺谱系 - 特异性TF HOXB13似乎在肿瘤组织特异性AR位置共定位。本提案的总体目标是表征肿瘤发生过程中AR重新编程的基本机制，以及 - 首次表征从局部前列腺癌发展到转移性，耐药性疾病的人类组织中AR程序的表征。在第一个目标中，将在细胞系模型中进行chip-seq，以表达AR的正常前列腺上皮（LHSAR）和前列腺癌（LNCAP和VCAP）。当每个TF通过shRNA和基因组编辑击倒或通过慢病毒转导过表达时，将测量AR，FOXA1和HOXB13的Cistromes的变化。还将针对每个细胞系条件进行RNA-seq，以确定受表观遗传重编程影响的基因。在第二和第三目标中，AR的表观遗传景观将通过人类规格中的芯片seq系统地绘制：局部组织，未处理的转移组织，耐Cantastration抗性转移酶和耐恩拉托胺耐药性疾病。该提案中概述的新实验的完成将无与伦比地观察主转录因子如何驱动前列腺癌的进展。具体而言，我们将发现：（i）在前列腺癌的发展和进展过程中如何重新编程AR，以及哪些共同调节器支持这一过程，（ii）在获得对依Zalutamide的抵抗力期间如何对AR进行重编程，以及（如果是这样），则在此过程中支持此过程，以及（III）其他非AR主管驱动癌症的癌症。提出的实验还将使我们能够识别受AR重编程影响的靶基因。随着疾病发展到耐Castration的转移状态，该项目将导致表观遗传景观的地图集，这是致命的。遗传基因座 完成该数据集的目标基因将刺激治疗干预的新目标。