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Genome-wide pan-haematopoietic transposon screening for oncogenic network discovery in mice

全基因组泛造血转座子筛选以发现小鼠致癌网络

基本信息

批准号：
386625870
负责人：
Professor Dr. Roland Rad
金额：
--
依托单位：
Institut für Molekulare Onkologie und Funktionelle Genomik
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2017
资助国家：
德国
起止时间：
2016-12-31 至 2020-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/386625870?language=en
关键词：
Genome wide pan haematopoietic transposon

项目摘要

Recent years have brought tremendous advances in the understanding of cancer genetics. Next generation sequencing efforts, for example, are creating catalogues of somatic mutations for principally all cancer types. Such novel insights are however also revealing that the complexity of molecular processes driving tumorigenesis is still far from being understood. For example, there is a lack of tools to systematically search for targets or effector pathways downstream of mutated human cancer genes, or to pinpoint cancer drivers that are not mutated but are among the thousands of epigenetically, transcriptionally or post-transcriptionally dysregulated genes in a cancer cell. To address these obstacles, we have developed PiggyBac transposon tools in mice and showed their application for in vivo genetic screening and cancer gene discovery. Using PiggyBac mouse lines specifically designed for screening in the haematopoietic system we were able to induce various types of haematologic malgnanices in pilot experiments, including T cell, B cell, myeloid or undifferentiated neoplasms and others. We propose to now extend this approach to perform systematic genome-wide surveys for cancer drivers in various types of heamatologic malignancies. Our pilot screens have already revealed novel cancer genes for some entities, including the splicing factor Mbnl1 in poorly differentiated cancers. Mbnl1 and few other selected novel cancer genes emerging from these screens will be validated and functionally characterized in vitro and in vivo using novel unpublished CRISPR/Cas9 vectors/mice developed in our lab. Our initial experiments using RNAi and CRISPR/Cas9-based gene editing in the Hoxb8 cell system of bone-marrow derived progenitor cells point towards a role of Mbnl1 in differentiation control, which will be explored further. Finally, cross-species mouse/human analyses will be performed using large leukaemia/lymphoma patient cohorts in order to establish the clinical relevance of novel candidate cancer genes. These genome-wide screens will discover cancer drivers that cannot be identified with other approaches to cancer genome analysis, and will thus complement the sequencing-based census of human cancer genes across haematologic malignancies.

近年来，对癌症遗传学的理解取得了巨大进展。例如，下一代测序工作正在为主要所有类型的癌症创建体细胞突变目录。然而，这些新颖的见解也揭示了驱动肿瘤发生的分子过程的复杂性仍远未被理解。例如，缺乏工具来系统地搜索突变的人类癌症基因下游的靶标或效应途径，或查明没有突变但在癌细胞中数千个表观遗传、转录或转录后失调基因中的癌症驱动因素。为了解决这些障碍，我们开发了小鼠PiggyBac转座子工具，并展示了它们在体内遗传筛选和癌症基因发现方面的应用。使用PiggyBac专门设计用于造血系统筛选的小鼠系，我们能够在中试实验中诱导各种类型的血液恶性肿瘤，包括T细胞，B细胞，骨髓或未分化肿瘤等。我们现在建议将这种方法扩展到对各种类型的血液恶性肿瘤的癌症驱动因素进行系统的全基因组调查。我们的试点筛选已经揭示了一些实体的新癌症基因，包括低分化癌症中的剪接因子Mbnl1。我们将利用实验室开发的新型未发表的CRISPR/Cas9载体/小鼠，在体外和体内验证Mbnl1和其他一些从这些筛选中出现的新型癌症基因并对其进行功能表征。我们在骨髓源性祖细胞Hoxb8细胞系统中使用RNAi和基于CRISPR/ cas9的基因编辑的初步实验表明Mbnl1在分化控制中的作用，将进一步探索。最后，将使用大型白血病/淋巴瘤患者队列进行跨物种小鼠/人类分析，以建立新的候选癌症基因的临床相关性。这些全基因组筛选将发现其他癌症基因组分析方法无法识别的癌症驱动因素，从而补充基于测序的人类恶性血液肿瘤癌症基因普查。