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

• 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

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genes emerging from these screens will be validated and functionally characterized in vitro and in vivo using novel unpublished CRISPR/Cas9 vectors/mice developed in our lab.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癌症类型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了解。例如，缺乏工具来系统地搜索突变人类癌症基因下游的靶点或效应通路，或者查明未突变但属于癌症中数千个表观遗传、转录或转录后失调基因的癌症驱动因素。癌细胞。为了解决这些障碍，我们在小鼠身上开发了 PiggyBac 转座子工具，并展示了它们在体内遗传筛查和癌症基因发现中的应用。使用专为造血系统筛选而设计的 PiggyBac 小鼠品系，我们能够在预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。我们现在建议扩展这种方法，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。 Mbnl1 和其他一些从这些筛选中选出的新型癌症基因将使用我们实验室开发的新型未发表的 CRISPR/Cas9 载体/小鼠在体外和体内进行验证和功能表征。我们在骨髓来源的祖细胞的 Hoxb8 细胞系统中使用 RNAi 和基于 CRISPR/Cas9 的基因编辑进行的初步实验表明了 Mbnl1 在分化控制中的作用，这将进一步探讨。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，以确定新候选癌症基因的临床相关性。这些全基因组筛查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，从而补充血液系统恶性肿瘤中基于测序的人类癌症基因普查。

项目成果

PiggyBac Transposon-Based Insertional Mutagenesis in Mice.

基于 PiggyBac 转座子的小鼠插入诱变。

• DOI: 10.1007/978-1-4939-8967-6_14
• 发表时间: 2018-12-13
• 期刊: Methods in molecular biology
• 作者: Mathias J Friedrich;I. Bronner;Pentao Liu;A. Bradley;R. Rad
• 通讯作者: R. Rad

Genetic alterations of the SUMO isopeptidase SENP6 drive lymphomagenesis and genetic instability in diffuse large B-cell lymphoma

SUMO 异构肽酶 SENP6 的遗传改变驱动淋巴瘤发生和弥漫性大 B 细胞淋巴瘤的遗传不稳定性

• DOI: 10.1038/s41467-021-27704-8
• 发表时间: 2022-01-12
• 期刊: Nature Communications
• 影响因子: 16.6
• 作者: Schick M;Zhang L;Maurer S;Maurer HC;Isaakaidis K;Schneider L;Patra U;Schunck K;Rohleder E;Hofstetter J;Baluapuri A;Scherger AK;Slotta-Huspenina J;Hettler F;Weber J;Engleitner T;Maresch R;Slawska J;Lewis R;Istvanffy R;Habringer S;Steiger K;Baiker A;Oostendorp RAJ;Miething C;Lenhof HP;Bassermann F;Chapuy B;Wirth M;Wolf E;Rad R;Müller S;Keller U
• 通讯作者: Keller U

Editorial overview: Functionalizing cancer genomes in the era of big data.

编辑概述：大数据时代的癌症基因组功能化

• DOI: 10.1016/j.gde.2019.06.011
• 发表时间: 2019
• 期刊: Current opinion in genetics & development
• 影响因子: 4
• 作者: Rad R; Boutros M
• 通讯作者: Boutros M

Engineering CRISPR mouse models of cancer.

改造 CRISPR 小鼠癌症模型。

• DOI: 10.1016/j.gde.2019.04.001
• 发表时间: 2019-02-01
• 期刊: Current opinion in genetics & development
• 影响因子: 4
• 作者: J. Weber;R. Rad
• 通讯作者: R. Rad

Genome-wide transposon screening and quantitative insertion site sequencing for cancer gene discovery in mice

全基因组转座子筛选和定量插入位点测序用于发现小鼠癌症基因

• DOI: 10.1038/nprot.2016.164
• 发表时间: 2017-01-12
• 期刊: Nature Protocols
• 影响因子: 14.8
• 作者: Mathias J Friedrich;Lena Rad;I. Bronner;Ale;er J. Strong;er;Wei Wang;J. Weber;Matthew Mayho;H. Ponstingl;T. Engleitner;C. Grove;A. Pfaus;D. Saur;J. Cadiñanos;M. Quail;G. Vassiliou;Pentao Liu;A. Bradley;R. Rad
• 通讯作者: R. Rad