Targeting oncogenic Myb fusions in salivary gland cancer with the elongation inhibitor SVC112

使用延伸抑制剂 SVC112 靶向唾液腺癌中的致癌 Myb 融合

基本信息

批准号：
10592292
负责人：
Antonio Jimeno
金额：
$ 49.79万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10592292
关键词：
ATP Hydrolysis Adenoid Cystic Carcinoma Affect Biochemical Biology CRISPR/Cas technology Cancer Cluster Cancer Model Cancer Patient Cancer cell line Catalogs Cell Line Cells Chimeric Proteins Clinical Clustered Regularly Interspaced Short Palindromic Repeats Colorado Complex Cyclin D1 Databases Dependence Disease Disease model Distant Metastasis Dose Drug Kinetics Event FDA approved FRAP1 gene Frequencies Gene Fusion Genes Genetic Genetic Markers Goals Growth Hour Human Hypoxia Implant Internal Ribosome Entry Site Invaded Lentivirus Vector Malignant Neoplasms Malignant neoplasm of salivary gland Measures Mediating Messenger RNA Modeling Molecular Molecular Target Mucoepidermoid Carcinoma Mus Mutate Mutation NFIB gene Oncogenic Orphan Other Genetics PIK3CG gene Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Physiological Population Predisposition Production Proliferating Property Protein Biosynthesis Protein Inhibition Protein Overexpression Protein Synthesis Inhibition Proteins Proteome Proteomics Radiation Rare Diseases Recurrence Reporting Ribosomes Role Seminal Signal Transduction Solid Neoplasm Stress Testing Therapeutic Therapeutic Index Toxic effect Transcript Translating Translations Tumorigenicity Unresectable Up-Regulation antitumor effect cancer cell clinically significant cohort exome sequencing gene product human model in vitro Assay in vitro Model in vivo in vivo Model in vivo evaluation inhibitor neoplastic cell novel novel therapeutics overexpression patient derived xenograft model pharmacologic predictive marker protein biomarkers ribosome profiling small molecule spectrograph targeted treatment transcription factor transcriptome sequencing tumor tumor behavior tumor growth tumor initiation tumorigenic

项目摘要

SUMMARY. Salivary gland cancer (SGC) is an orphan disease for which no targeted therapies are approved. SGCs are divided into histotypes, the most common being adenoid cystic carcinoma (ACC) and mucoepidermoid carcinoma (MEC). We have generated one of the largest reported SGC PDX banks including major histotypes like ACC and the first reported PDX models of MEC. Our SGC PDX bank includes several cases from the same patient, collected after subsequent relapses, which has allowed an exploration of the acquisition of oncogenic gene events. Similar to other reports, our SGC models had low mutation burden by whole exome sequencing, and RNA-seq analysis identified known (MYB-NFIB), novel (NFIB-MTFR2), and even dual gene fusions in ACC cases that gave rise to oncogenic fusion protein products. Other genetic events in SGC clustered in the PI3K and mTOR pathways, which impinge upon protein synthesis. Both MEC and ACC overexpress key transcription factors such as Myc or SOX2 that dictated tumorigenicity and growth in SGC. Overall, we propose that protein synthesis is an unexplored target in SGC. SVC112 is a small molecule that inhibits protein synthesis at the elongation step by inhibiting eEF2. In our seminal studies where SVC112 was first reported, protein synthesis and growth inhibition were associated; SVC112 had greater effect on cancer over non-cancer cells; SVC112 depleted SOX2, Myc, and Cyclin D1, and arrested growth in vivo. In both ACC and MEC cell lines we found that SVC112 inhibits both native Myb and the protein products of MYB fusions and key proteins like Myc. SVC112 inhibited proliferation and sphere formation in both ACC and MEC in vitro assays, and had notable single agent activity (including actual tumor shrinkage) in 2 ACC PDX, one with a MYB fusion and another one showing non- fusion mediated Myb upregulation. We propose to study the role of oncogenic fusions in SGC, the mechanism of action of SVC112 in SGC, and the efficacy of SVC112 in complex SGC models. First, we will catalogue and prioritize MYB fusions from large patient and PDX cohorts. Then, to understand key genetic events we will insert fusions in non-cancer cell lines, and deploy CRISPR and lentiviral vectors in fusion-and non-fusion-bearing SGC cell lines, respectively; we will also test SVC112 inhibition to add an additional layer of testing of the impact of protein modulation. The hypothesis that SVC112’s selective effect is due to selective depletion of key proteins will be tested by ribosome profiling (to identify mRNA targets) and proteomics analysis (to identify proteins targets). Lastly, we will test the in vivo efficacy of SVC112 using both native SGC PDX models bearing MYB and other fusions and wild-tpy, as well as in vivo models with the CRISPR-depleted MYB fusions strains. Overall our goal is to examine dependence on fusion events vs protein overexpression by other molecular mechanisms for SVC112 susceptibility. This project will propel the translation of SVC112, a new drug for SGC discovered in Colorado, by dissecting the basis for its effect and therapeutic window, and identifying its efficacy in advanced SGC models with a plan that enables identification of which subset of SGC patients may derive more benefit.

概括。唾液腺癌（SGC）是一种未批准靶向疗法的孤儿疾病。 SGC分为组织型，最常见的是腺样囊性癌（ACC）和粘膜外皮样癌（MEC）。我们已经生成了报告的最大的SGC PDX银行之一，包括主要的组织型像ACC和第一个报告的MEC的PDX模型一样。我们的SGC PDX银行包括来自同一案例患者，随后的继电器后收集，这允许探索对致癌性的获取基因事件。与其他报告类似，我们的SGC模型通过整个外显子组测序的突变伯嫩低， RNA-seq分析已鉴定出已知（MYB-NFIB），新颖（NFIB-MTFR2），甚至是ACC中的双基因融合引起致癌融合蛋白产物的病例。 SGC中的其他遗传事件聚集在PI3K中和MTOR途径，会影响蛋白质合成。 MEC和ACC过表达密钥转录诸如SGC中肿瘤性和生长的MYC或SOX2之类的因素。总体而言，我们提出了该蛋白质合成是SGC中意外的目标。 SVC112是一个小分子，可抑制蛋白质合成通过抑制EEF2进行伸长一步。在我们的第二项研究中，首先报道了SVC112，蛋白质合成与生长抑制相关； SVC112对非癌细胞的癌症影响更大。 SVC112 Sox2，Myc和Cyclin D1耗尽，并在体内捕捉生长。在ACC和MEC细胞系中，我们发现 SVC112抑制了本地MYB和MYB融合的蛋白质产品和MyC（如MYC）的关键蛋白质。 SVC112 在体外测定中抑制ACC和MEC中的增殖和球体形成，并具有显着的单药在2个ACC PDX中的活性（包括实际肿瘤收缩），一个具有MYB融合融合介导的MYB上调。我们建议研究致癌融合在SGC（机制）中的作用 SVC112在SGC中的作用，以及复杂SGC模型中SVC112的效率。首先，我们将分类和优先考虑大型患者和PDX队列的MYB融合。然后，要了解关键的遗传事件，我们将插入非癌细胞系中的融合，以及在融合和非融合的SGC中部署CRISPR和慢病毒载体细胞系分别；我们还将测试SVC112抑制作用，以增加对影响的额外测试层蛋白质调节。 SVC112的选择性作用是由于关键蛋白的选择性耗竭引起的假设将通过核糖体分析（以识别mRNA靶标）和蛋白质组学分析来测试（鉴定蛋白质目标）。最后，我们将使用带有MYB和MYB的本机SGC PDX模型来测试SVC112的体内效率其他融合和野生型，以及带有CRISPR融合的MYB融合菌株的体内模型。总体而言目标是通过其他分子机制来检查对融合事件与蛋白过表达的依赖 SVC112敏感性。该项目将推动SVC112的翻译，这是一种针对SGC的新药。科罗拉多州，通过剖析其效果和治疗窗口的基础，并确定其在高级中的有效具有计划的SGC模型可以识别哪些SGC患者子集可能会获得更多的好处。