CALR frameshift mutations in MPN patient-derived iPSCs accelerate maturation of megakaryocytes.
CALR frameshift mutations in MPN patient-derived iPSCs accelerate maturation of megakaryocytes.
复制标题
DOI：
10.1016/j.stemcr.2021.09.019
复制
发表时间：
2021-11-09
影响因子：
5.9
通讯作者：
Koschmieder S
中科院分区：
文献类型：
--
作者：
Olschok K;Han L;de Toledo MAS;Böhnke J;Graßhoff M;Costa IG;Theocharides A;Maurer A;Schüler HM;Buhl EM;Pannen K;Baumeister J;Kalmer M;Gupta S;Boor P;Gezer D;Brümmendorf TH;Zenke M;Chatain N;Koschmieder S
钙网蛋白(CALR)突变是骨髓增生性肿瘤(MPN)的驱动突变，导致血小板生成素受体激活，导致异常巨核细胞生成。在这里，我们产生了患者来源的CALRins5或CALRdel52阳性的诱导多能干细胞(IPSCs)，以建立用于分子和机制研究的MPN疾病模型。我们证明了来自纯合CALR突变的IPSCs的粒细胞中存在髓过氧化物酶缺乏症，通过使用CRISPR/Cas9修复突变而被挽救。IPSC来源的巨核细胞具有原代巨核细胞的特征，如形成分界膜系统和胞浆前血小板突起。重要的是，CALR突变导致巨核细胞生成增强，并以不依赖于血小板生成素的方式加速巨核细胞的发育。从机制上讲，我们的研究确定了突变的巨核细胞和未突变的巨核细胞的不同调控途径，如缺氧信号，这代表了治疗干预的潜在靶点。总之，我们证明了突变的CALR驱动的致病机制的关键方面取决于其合子，并发现了新的治疗靶点，使我们的模型成为MPNS临床药物筛选的有价值的工具。CALR突变的IPSCs可以有效地对人类MPN疾病进行建模CRISPR介导的CALR突变的修复挽救了正常的IPSC功能CALR突变的IPSCs中的巨核生成是增殖和加速转录在突变的巨核细胞的筛选确定新的治疗方案在本文中，Koschmieder及其同事建立了一种新的MPN患者专用的IPSC模型，以研究钙网蛋白(CALR)移码突变在造血中的影响。他们报告了由CALR突变引起的增生和加速的巨核细胞生成。IPSC来源的巨核细胞的RNA测序突出了突变和未突变的巨核细胞之间的转录差异，例如低氧相关途径的上调。
Calreticulin (CALR) mutations are driver mutations in myeloproliferative neoplasms (MPNs), leading to activation of the thrombopoietin receptor and causing abnormal megakaryopoiesis. Here, we generated patient-derived CALRins5- or CALRdel52-positive induced pluripotent stem cells (iPSCs) to establish an MPN disease model for molecular and mechanistic studies. We demonstrated myeloperoxidase deficiency in granulocytic cells derived from homozygous CALR mutant iPSCs, rescued by repairing the mutation using CRISPR/Cas9. iPSC-derived megakaryocytes showed characteristics of primary megakaryocytes such as formation of demarcation membrane system and cytoplasmic pro-platelet protrusions. Importantly, CALR mutations led to enhanced megakaryopoiesis and accelerated megakaryocytic development in a thrombopoietin-independent manner. Mechanistically, our study identified differentially regulated pathways in mutated versus unmutated megakaryocytes, such as hypoxia signaling, which represents a potential target for therapeutic intervention. Altogether, we demonstrate key aspects of mutated CALR-driven pathogenesis dependent on its zygosity, and found novel therapeutic targets, making our model a valuable tool for clinical drug screening in MPNs. CALR-mutated iPSCs allow efficient modeling of human MPN disease CRISPR-mediated repair of CALR mutations rescues normal iPSC function Megakaryopoiesis in CALR-mutated iPSCs is hyperplastic and accelerated Transcriptome screen of mutated megakaryocytes identifies novel therapeutic options In this article, Koschmieder and colleagues establish a novel MPN patient-specific iPSC model to investigate the impact of calreticulin (CALR) frameshift mutations in hematopoiesis. They report hyperplastic and accelerated megakaryopoiesis caused by CALR mutations. RNA sequencing of iPSC-derived megakaryocytes highlights transcriptomic differences between mutated and unmutated megakaryocytes, such as upregulation of hypoxia-related pathways.
登录
查看更多内容
影响因子：
12.8
作者：
Barbui T;Thiele J;Gisslinger H;Kvasnicka HM;Vannucchi AM;Guglielmelli P;Orazi A;Tefferi A
通讯作者：
Tefferi A
影响因子：
21.3
作者：
Decker M;Martinez-Morentin L;Wang G;Lee Y;Liu Q;Leslie J;Ding L
通讯作者：
Ding L
影响因子：
28.2
作者：
Elf S;Abdelfattah NS;Chen E;Perales-Patón J;Rosen EA;Ko A;Peisker F;Florescu N;Giannini S;Wolach O;Morgan EA;Tothova Z;Losman JA;Schneider RK;Al-Shahrour F;Mullally A
通讯作者：
Mullally A
影响因子：
6
作者：
Liu, Yanfeng;Wang, Ying;Wang, Zack Z.
通讯作者：
Wang, Zack Z.
影响因子：
158.5
作者：
Klampfl, Thorsten;Gisslinger, Heinz;Kralovics, Robert
通讯作者：
Kralovics, Robert