Humanized NSG mouse model to study combinatorial leukemogenic effects of inherited ELANE and acquired CSF3R/RUNX1 mutations in congenital neutropenia

人源化 NSG 小鼠模型，用于研究先天性中性粒细胞减少症中遗传性 ELANE 和获得性 CSF3R/RUNX1 突变的组合致白血病效应

基本信息

批准号：
290677262
负责人：
Professorin Dr. Julia Skokowa, Ph.D.
金额：
--
依托单位：
Klinik für Hals-Nasen-Ohrenheilkunde
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2016
资助国家：
德国
起止时间：
2015-12-31 至 2020-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/290677262?language=en
关键词：
Humanized NSG mouse model study

项目摘要

Severe congenital neutropenia (CN) is a pre-leukemic bone marrow failure syndrome with a high risk of evolving into leukemia or myelodysplastic syndrome (MDS). Recently we demonstrated a very high frequency of cooperating RUNX1 and CSF3R mutations in CN patients who developed leukemia or MDS. CSF3R mutations alone are unable to induce leukemia in CN patients or in mice expressing a transgenic mutated G-CSFR (d715) and we hypothesized, that co-acquisition of RUNX1 on the background of inherited ELANE mutations is an essential step in the leukemogenic transformation in CN. Moreover, we recently found that 27 % of ELANE-mutated CN patients who developed leukemia harbor same ELANE mutation at the position p.C151Y. Interestingly, sequential analysis of the gene expression signature of hematopoietic cells of CN patients who harbors C151Y ELANE mutation and developed leukemia revealed rapid transformation of CSF3R-mutated hematopoietic progenitors into more primitive hematopoietic progenitors after co-acquisition of RUNX1 mutation. Based on these observations, we hypothesized, that co-acquisition of RUNX1 and CSF3R mutations on the pathological background due to inheritance of ELANE mutations, especially C151Y mutation, shifts the hematopoietic differentiation program towards more primitive hematopoietic progenitors with elevated proliferative capacity and reduced myeloid differentiation, which ultimately lead to leukemia. In the present study we aimed to further investigate the role of cooperative inherited ELANE and acquired CSF3R and RUNX1 mutations on the leukemogenic transformation of HSCs in humanized mouse model in vivo. We plan to study the importance of the time sequence of acquisition of RUNX1 and CSF3R mutations in leukemogenic transformation and the role of inherited CN-specific ELANE mutation (C151Y) in this process. We also plan to delineate leukemogenic intracellular signal transduction pathway/s activated in HSCs in the presence of ELANE, CSF3R and RUNX1 mutations. Our long-term aim is to establish methods of correction of the leukemogeinc factors/signaling pathways using small molecules in order to prevent or to treat leukemia in CN patients harboring ELANE mutations.

严重的先天性中性粒细胞减少症（CN）是一种美质的骨髓衰竭综合征，具有高风险，即变成白血病或骨髓增生性塑料综合征（MDS）。最近，我们证明了患有白血病或MDS的CN患者的RUNX1和CSF3R突变的频率很高。仅CSF3R突变就无法诱导CN患者或表达转基因突变G-CSFR（D715）的小鼠中的白血病（D715），我们假设，Runx1在遗传的Elane突变的背景下共同获得Runx1在CN中的白血病转化中是必不可少的步骤。此外，我们最近发现，在P.C151Y位置出现了白血病相同的岩岩突变的Elane-Muthated CN患者。有趣的是，对C151Y Elane突变和发展性白血病的CN患者的基因表达信号的顺序分析表明，CSF3R突变的造血祖细胞快速转化为更具原始的造血祖细胞后，是在Runx1突变共同的造血祖细胞。基于这些观察，我们假设，由于梯形突变的遗传，尤其是C151Y突变，Runx1和CSF3R突变在病理背景上的共同出现，将造血分化程序转移到更原始的造血祖细胞上，而造血祖细胞具有更高的能力和降低的基因差异，并导致了我的升级和降低，这是较终的造成物质。在本研究中，我们旨在进一步研究合作遗传的Elane的作用，并获得了人体化小鼠模型中HSC的白血病转化中CSF3R和RUNX1突变的作用。我们计划研究runx1和CSF3R突变在白血病转化中获取的时间顺序的重要性，以及在此过程中遗传的CN特异性基质突变（C151Y）的作用。我们还计划在存在Elane，CSF3R和Runx1突变的情况下在HSC中激活的细胞内信号转导途径。我们的长期目的是建立使用小分子校正白血病因子/信号通路的方法，以预防或治疗具有浮力突变的CN患者中的白血病。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Ultra-Sensitive CSF3R Deep Sequencing in Patients With Severe Congenital Neutropenia

DOI：
10.3389/fimmu.2019.00116
发表时间：
2019-02-28
期刊：
FRONTIERS IN IMMUNOLOGY
影响因子：
7.3
作者：
Klimiankou, Maksim;Uenalan, Murat;Welte, Karl
通讯作者：
Welte, Karl

Gene Knockout in Hematopoietic Stem and Progenitor Cells Followed by Granulocytic Differentiation.

造血干细胞和祖细胞中的基因敲除随后进行粒细胞分化

DOI：
10.1007/978-1-0716-0290-4_26
发表时间：
2020
期刊：
Methods in molecular biology
影响因子：
0
作者：
Ritter M;Welte K;Skokowa J;Klimiankou M
通讯作者：
Klimiankou M

CRISPR/Cas9-mediated ELANE knockout enables neutrophilic maturation of primary hematopoietic stem and progenitor cells and induced pluripotent stem cells of severe congenital neutropenia patients

CRISPR/Cas9介导的ELANE敲除使严重先天性中性粒细胞减少症患者的原代造血干细胞和祖细胞以及诱导多能干细胞的中性粒细胞成熟

DOI：
10.3324/haematol.2019.221804
发表时间：
2020
期刊：
Haematologica
影响因子：
10.1
作者：
Nasri M;Ritter M;Dannenmann B;Aghaallaei N;Amend D;Makaryan V;Fletcher B;Bernhard R;Steiert I;Hahnel K;Berger J;Koch I;Sailer B;Hipp K;Zeidler C;Klimiankou M;Bajoghli B;Dale DC;Welte K;Skokowa J
通讯作者：
Skokowa J

New insights into the pathomechanism of cyclic neutropenia