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

• 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

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.

重度先天性中性粒细胞减少症（CN）是一种白血病前期骨髓衰竭综合征，具有演变为白血病或骨髓增生异常综合征（MDS）的高风险。最近，我们证明了一个非常高的频率合作RUNX 1和CSF 3R突变的CN患者谁发展白血病或MDS。单独的CSF 3R突变不能在CN患者或表达转基因突变G-CSFR（d 715）的小鼠中诱导白血病，并且我们假设，在遗传ELANE突变的背景下共同获得RUNX 1是CN中致白血病转化的重要步骤。此外，我们最近发现，27%的ELANE突变的CN患者发生白血病，在p.C151Y位置具有相同的ELANE突变。有趣的是，对携带C151 Y ELANE突变并发生白血病的CN患者的造血细胞的基因表达特征的序列分析揭示了在共获得RUNX 1突变后，CSF 3R突变的造血祖细胞快速转化为更原始的造血祖细胞。基于这些观察结果，我们假设，由于ELANE突变，特别是C151 Y突变的遗传，在病理背景上共同获得RUNX 1和CSF 3R突变，使造血分化程序向具有升高的增殖能力和降低的髓样分化的更原始的造血祖细胞转移，这最终导致白血病。在本研究中，我们的目的是进一步研究协同遗传ELANE和获得性CSF 3R和RUNX 1突变对人源化小鼠模型体内HSC白血病转化的作用。我们计划研究在白血病转化中获得RUNX 1和CSF 3R突变的时间顺序的重要性以及遗传CN特异性ELANE突变（C151 Y）在此过程中的作用。我们还计划描绘在ELANE、CSF 3R和RUNX 1突变存在下HSC中激活的致白血病细胞内信号转导通路。我们的长期目标是建立使用小分子校正白血病原因子/信号通路的方法，以预防或治疗携带ELANE突变的CN患者的白血病。

项目成果

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
• 作者: Ritter M;Welte K;Skokowa J;Klimiankou M
• 通讯作者: Klimiankou M

New insights into the pathomechanism of cyclic neutropenia

• DOI: 10.1111/nyas.14309
• 发表时间: 2020-02-21
• 期刊: ANNALS OF THE NEW YORK ACADEMY OF SCIENCES
• 影响因子: 5.2
• 作者: Mir, Perihan;Klimiankou, Maksim;Welte, Karl
• 通讯作者: Welte, Karl

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