Role of newly identified, thyroid-specific LincRNA in BRAFV600E thyroid carcinoma

新发现的甲状腺特异性 LincRNA 在 BRAFV600E 甲状腺癌中的作用

基本信息

批准号：
10368959
负责人：
Carmelo Nucera
金额：
$ 39.23万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10368959
关键词：
3-Dimensional ATAC-seq Affect Antisense RNA Binding Sites Biology CDK4 gene Cancer Patient Cell Death Cell Proliferation Cell Survival Cells Chromatin Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Code Combined Modality Therapy Complex DNA Binding Data Disease Progression Down-Regulation Drug resistance E2F transcription factors ERBB3 gene EZH2 gene Early Diagnosis Enhancers Enzymes Epigenetic Process Extracellular Matrix Proteins FRAP1 gene Genes Genetic Transcription Genomic Instability Histone Acetylation Histones Human Hydrogen Peroxide In Vitro Iodide Peroxidase Iodine Link Malignant Neoplasms Maps Metabolism Mutation Neoplasm Metastasis Normal tissue morphology Oxidative Stress Papillary thyroid carcinoma Paracrine Communication Pathway interactions Patient-Focused Outcomes Pericytes Pharmacotherapy Play Proliferating RNA Recurrence Regulation Reporting Resistance Resistance development Role Sampling Secondary to Signal Transduction Survival Rate Technology Therapeutic Thrombospondin 1 Thyroid Gland Thyroid carcinoma Time Tissues Transcription Repressor Untranslated RNA Up-Regulation XCL1 gene cancer cell clinically significant density design drug development epigenetic silencing histone methylation in vivo inhibitor mortality mouse model neoplastic cell novel novel therapeutics overexpression promoter radioiodine therapy restoration screening synergism targeted treatment transcriptome transcriptome sequencing translational impact tumor tumor growth tumor microenvironment tumor progression uptake

项目摘要

BRAFV600E-papillary thyroid carcinoma (V600E-PTC) is particularly aggressive, and as it deregulates iodine metabolism, its victims are unresponsive to radioiodine treatment, and suffer high rates of metastasis and recurrence, and low survival rates. BRAFV600E inhibitors such as vemurafenib (vemu), which at first showed clinical promise, are the only therapeutic option, but resistance to these drugs is widely reported. Our premise is that the epigenetic silencing of the first identified thyroid-specific long intergenic non-coding RNA (lincRNA), Xloc001313 (Xloc13) deregulates iodine metabolism, sustains tumor cell survival, and promotes progression and drug resistance. Xloc13 is active downstream of the coding gene for TPO, a key enzyme for iodine metabolism, and its expression is far lower in V600E-PTC than normal tissue. Our project is designed to determine the cellular mechanisms behind these effects, and use these findings to better target clinical trials of new treatments. Vemu treatment induces Xloc13 expression, indicating it is silenced by BRAFV600E, and its inactivation by CRISPR in normal thyroid cells reduces TPO levels and iodine uptake, while increasing TSP-1 levels (extracellular matrix protein) and cell proliferation. Xloc13 RNA enhancer maps to DNA-binding motifs on the TSP-1 promoter, leading to down-regulation of TSP-1, which inactivates TGFβ1 to inhibit PTC cell and pericyte viability. We recently demonstrated that V600E-PTC samples are enriched in pericytes that co- express TSP-1 and TGFβ1 to sustain V600E-PTC cell survival. Xloc13 silencing correlates with high pericyte density, linking its action to pericyte biology. We also identified regulatory motifs in the Xloc13 promoter for EZH2 and E2F1 as transcriptional repressors up-regulated in vemu-resistant V600E-PTC cells and ChIP- enriched at these DNA-binding motifs. Critically, combining inhibitors of BRAFV600E+EZH2+CDK4/6 durably rescued Xloc13, and reduced PTC cell and pericyte viability. Xloc13 down-regulation therefore plays a key role in V600E-PTC, and our central objective is to determine its mechanisms of regulation and their impact on tumor progression and resistance. We hypothesize that BRAFV600E synergizes with EZH2 and the CDK4/6 (upstream of E2F1) pathway to down-regulate Xloc13, which deregulates iodine metabolism, enhances tumor viability, and increases invasion signaling through pericyte action, which leads to secondary drug resistance and tumor progression. Using cutting-edge technologies (CRISPR, ChIP, ATAC, RAP) and new cancer mouse model, our specific Aims are: 1) To identify the mechanisms regulating Xloc13 in V600E-PTC and their impact on tumor aggressiveness; 2) To determine the role of Xloc13 in the regulation of TPO in V600E-PTC and its impact on iodine metabolism and PTC cell survival; 3) To assess the efficacy of anti-BRAFV600E-EZH2-CDK4/6 combined therapy on human V600E-PTC in vivo. Our studies will have high clinical significance by supporting new clinical trials of trimodal combined therapies for advanced V600E-PTC patients, and identifying new mechanisms in PTC and its microenvironment which could serve as drug development targets for this tumor.

BRAFV600E毛细血管甲状腺癌（V600E-PTC）特别有侵略性，并且由于它消除了碘代谢对放射性碘治疗，并且遭受了较高的转移和复发率，并且最初表现出临床前景。，是唯一的故事，但是这些药物被广泛报道。进度和耐药性。新的新新的新新的新新的新新的新新的新新的新新的新新的新新型新的新TS。蛋白质）和cel序言。表示维持V600E-PTC细胞存活。密度，我们还鉴定了XLOC13启动子中的EZH2启动子的调节基序，并在抗Vemu的V600E-PTC细胞中上调，并在这些DNA结合基序中富含芯片/6持续的XLOC13，PTC和周细胞生存力降低。 EZH2和CDK4/6（E2F1的上游）OC13，它消除了碘代谢，增强肿瘤生存力，并增加了在周围的耐药性耐药性和肿瘤进展的范围内的发明信号。肿瘤敏捷性； 2）确定XLOC13在TPO中的作用，在V600E对碘代谢和PTC细胞存活中的影响；山丘高临床意义，通过支持三摩德疗法的新临床临床疗法，用于晚期V600E-PTC Ients，并在PTC及其微环境中进行了新机制，该机制可以用作Servd Servd Servd Servd作为开发TAR TAR TARGERGERES肿瘤。