Modelling the erythroblastic island niche of dyserythropoietic anaemia type IV patients using induced pluripotent stem cells.
Modelling the erythroblastic island niche of dyserythropoietic anaemia type IV patients using induced pluripotent stem cells.
复制标题
DOI：
10.3389/fcell.2023.1148013
复制
发表时间：
2023
影响因子：
5.5
通讯作者：
Forrester, Lesley M.
中科院分区：
文献类型：
--
作者：
May, Alisha;Ventura, Telma;Fidanza, Antonella;Volmer, Helena;Taylor, Helen;Romano, Nicola;D'Souza, Sunita L.;Bieker, James J.;Forrester, Lesley M.
关键词：
简介：IV 型先天性红细胞生成障碍性贫血 (CDA) 与转录因子 KLF1 中的氨基酸取代 Glu325Lys (E325K) 有关。这些患者表现出一系列症状，包括外周血中持续存在有核红细胞 (RBC)，这反映了 KLF1 在红细胞谱系中的已知作用。红细胞成熟和去核的最后阶段发生在与 EBI 巨噬细胞密切相关的成红细胞岛 (EBI) 生态位内。目前尚不清楚 KLF1 中 E325K 突变的有害影响是否仅限于红细胞谱系，或者与其生态位相关的巨噬细胞的缺陷是否也会导致疾病病理。 方法：为了解决这个问题，我们使用来自一名 CDA IV 型患者的诱导多能干细胞 (iPSC) 以及两个经过基因改造以表达可被 4OH-他莫昔芬激活的 KLF1-E325K-ERT2 蛋白的 iPSC 系，生成了人类 EBI 生态位的体外模型。将一名患者 iPSC 系与来自两名健康供体的对照系进行比较，并将 KLF1-E325K-ERT2 iPSC 系与从同一亲本 iPSCS 生成的一种诱导型 KLF1-ERT2 系进行比较。 结果：CDA 患者来源的 iPSC 和表达激活的 KLF1-E325K-ERT2 蛋白的 iPSC 在红细胞生成方面表现出显着缺陷，并伴有一些已知 KLF1 靶基因的破坏。所有 iPSC 系均可产生巨噬细胞，但当 E325K-ERT2 融合蛋白被激活时，我们注意到生成了由 CD93 标记的稍微不太成熟的巨噬细胞群。支持红细胞去核能力下降的微妙趋势也与携带 E325K-ERT2 转基因的巨噬细胞有关。 讨论：总的来说，这些数据支持这样的观点，即 KLF1-E325K 突变的临床显着影响主要与红系谱系缺陷相关，但生态位缺陷可能有可能加剧病情。我们描述的策略提供了一种强大的方法来评估 KLF1 中其他突变以及与 EBI 生态位相关的其他因素的影响。
Introduction: Congenital dyserythropoietic anaemia (CDA) type IV has been associated with an amino acid substitution, Glu325Lys (E325K), in the transcription factor KLF1. These patients present with a range of symptoms, including the persistence of nucleated red blood cells (RBCs) in the peripheral blood which reflects the known role for KLF1 within the erythroid cell lineage. The final stages of RBCs maturation and enucleation take place within the erythroblastic island (EBI) niche in close association with EBI macrophages. It is not known whether the detrimental effects of the E325K mutation in KLF1 are restricted to the erythroid lineage or whether deficiencies in macrophages associated with their niche also contribute to the disease pathology. Methods: To address this question, we generated an in vitro model of the human EBI niche using induced pluripotent stem cells (iPSCs) derived from one CDA type IV patient as well as two iPSC lines genetically modified to express an KLF1-E325K-ERT2 protein that could be activated with 4OH-tamoxifen. The one patient iPSC line was compared to control lines from two healthy donors and the KLF1-E325K-ERT2 iPSC line to one inducible KLF1-ERT2 line generated from the same parental iPSCS. Results: The CDA patient-derived iPSCs and iPSCs expressing the activated KLF1-E325K-ERT2 protein showed significant deficiencies in the production of erythroid cells with associated disruption of some known KLF1 target genes. Macrophages could be generated from all iPSC lines but when the E325K-ERT2 fusion protein was activated, we noted the generation of a slightly less mature macrophage population marked by CD93. A subtle trend in their reduced ability to support RBC enucleation was also associated with macrophages carrying the E325K-ERT2 transgene. Discussion: Taken together these data support the notion that the clinically significant effects of the KLF1-E325K mutation are primarily associated with deficiencies in the erythroid lineage but it is possible that deficiencies in the niche might have the potential to exacerbate the condition. The strategy we describe provides a powerful approach to assess the effects of other mutations in KLF1 as well as other factors associated with the EBI niche.
登录
查看更多内容
影响因子：
64.5
作者：
MADTES, DK;RAINES, EW;ROSS, R
通讯作者：
ROSS, R
影响因子：
46.9
作者：
Hockemeyer, Dirk;Soldner, Frank;Beard, Caroline;Gao, Qing;Mitalipova, Maisam;DeKelver, Russell C.;Katibah, George E.;Amora, Ranier;Boydston, Elizabeth A.;Zeitler, Bryan;Meng, Xiangdong;Miller, Jeffrey C.;Zhang, Lei;Rebar, Edward J.;Gregory, Philip D.;Urnov, Fyodor D.;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf
影响因子：
82.9
作者：
通讯作者：
--
影响因子：
4
作者：
Bernecker, Claudia;Ackermann, Mania;Dorn, Isabel
通讯作者：
Dorn, Isabel
影响因子：
5.4
作者：
El-Battrawy I;Lan H;Cyganek L;Zhao Z;Li X;Buljubasic F;Lang S;Yücel G;Sattler K;Zimmermann WH;Utikal J;Wieland T;Ravens U;Borggrefe M;Zhou XB;Akin I
通讯作者：
Akin I