A novel prospective isolation of murine fetal liver progenitors to study in utero hematopoietic defects.
A novel prospective isolation of murine fetal liver progenitors to study in utero hematopoietic defects.
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DOI：
10.1371/journal.pgen.1007127
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发表时间：
2018-01
期刊：
影响因子：
4.5
通讯作者：
Lacaud G
中科院分区：
文献类型：
--
作者：
Draper JE;Sroczynska P;Fadlullah MZH;Patel R;Newton G;Breitwieser W;Kouskoff V;Lacaud G
近年来，已经实现了对成人骨髓（BM）髓系祖细胞的高度详细的表征，因此，可以精确地确定体细胞缺陷对不同造血谱系命运决定的影响。胎儿肝脏（FL）造血祖细胞（HPC）的特点是比较差，潜在地阻碍了遗传变异对妊娠中期造血的影响的研究。许多疾病，例如婴儿急性白血病，具有子宫内起源，因此它们的研究将受益于分离高度纯化的祖细胞亚群的能力。我们先前证明Runx 1远端启动子（P1）-GFP：：近端启动子（P2）-hCD 4双报告基因小鼠（小家鼠）模型可用于鉴定具有不同谱系偏好的成年BM祖细胞亚群。在这项研究中，我们进行了FL中Runx 1-P1-GFP和P2-hCD 4表达的表征。FL免疫表型巨核细胞-红系祖细胞（MEP）和共同髓系祖细胞（CMP）隔室中P2-hCD 4的表达对应于粒细胞/单核细胞/巨核细胞的增加和红系特异性的减少。此外，Runx 1-P2-hCD 4表达与几种内源性细胞表面标志物的表达相关，包括CD 31和CD 45，为在转基因小鼠模型中前瞻性地鉴定高度纯化的胎儿髓系祖细胞提供了新的策略。我们利用这种方法来比较单独缺失总RUNX 1或RUNX 1C的影响，并确定受影响最大的胎儿HPC谱系。因此，FL祖细胞的这种新的前瞻性鉴定提高了以比以前更高的精度鉴定潜在基因网络的前景。红细胞、产生血小板的巨核细胞以及免疫应答导向的粒细胞和单核细胞的产生在发育中的胚胎的早期阶段开始，并在整个生命过程中持续。每种细胞类型的比例取决于生物体的特定需求。我们知道，在小鼠胚胎中，特化的血液祖细胞出现在胎儿肝脏中，并产生成熟的血细胞以响应不同的信号。然而，很难以足够高的准确度区分红细胞和白色血细胞生成祖细胞以研究这些线索。例如，我们知道几种儿童血液疾病，如白血病，是由出生前血液祖细胞的基因突变引起的，但如果我们不知道收集哪种血液祖细胞，那么在小鼠疾病模型中研究这些突变的影响就会受到阻碍。我们使用了不同的遗传标记来帮助区分红细胞，巨核细胞和产生粒细胞/单核细胞的祖细胞，其精确度比以前更高。此外，为了说明这种技术如何用于研究血液疾病，我们证明了影响转录因子Runx 1的突变会损害不同祖细胞以不同方式产生成熟血细胞的能力。
In recent years, highly detailed characterization of adult bone marrow (BM) myeloid progenitors has been achieved and, as a result, the impact of somatic defects on different hematopoietic lineage fate decisions can be precisely determined. Fetal liver (FL) hematopoietic progenitor cells (HPCs) are poorly characterized in comparison, potentially hindering the study of the impact of genetic alterations on midgestation hematopoiesis. Numerous disorders, for example infant acute leukemias, have in utero origins and their study would therefore benefit from the ability to isolate highly purified progenitor subsets. We previously demonstrated that a Runx1 distal promoter (P1)-GFP::proximal promoter (P2)-hCD4 dual-reporter mouse (Mus musculus) model can be used to identify adult BM progenitor subsets with distinct lineage preferences. In this study, we undertook the characterization of the expression of Runx1-P1-GFP and P2-hCD4 in FL. Expression of P2-hCD4 in the FL immunophenotypic Megakaryocyte-Erythroid Progenitor (MEP) and Common Myeloid Progenitor (CMP) compartments corresponded to increased granulocytic/monocytic/megakaryocytic and decreased erythroid specification. Moreover, Runx1-P2-hCD4 expression correlated with several endogenous cell surface markers’ expression, including CD31 and CD45, providing a new strategy for prospective identification of highly purified fetal myeloid progenitors in transgenic mouse models. We utilized this methodology to compare the impact of the deletion of either total RUNX1 or RUNX1C alone and to determine the fetal HPCs lineages most substantially affected. This new prospective identification of FL progenitors therefore raises the prospect of identifying the underlying gene networks responsible with greater precision than previously possible. The production of red blood cells, platelet-producing megakaryocytes, and immune response-directing granulocytes and monocytes is initiated at an early stage in the developing embryo and continues throughout life. The proportion of each cell type varies depending on the specific needs of the organism. We know that in the mouse embryo, specialized blood progenitor cells emerge in the fetal liver and produce mature blood cells in response to different cues. However, it is difficult to distinguish between red blood cell and white blood cell-producing progenitors with sufficiently high accuracy to study these cues. For example, we know that several childhood blood disorders, such as leukemias, are caused by genetic mutations in blood progenitor cells before birth, but studying the effects of these mutations in a mouse disease model is hampered if we don’t know which blood progenitor cells to collect. We have used different genetic markers to help distinguish red blood cell, megakaryocyte and granulocyte/monocyte-producing progenitor cells with a greater precision than was previously possible. Furthermore, to illustrate how this technique can be used to study blood disorders, we demonstrated that mutations affecting the transcription factor Runx1 impair the abilities of different progenitors to produce mature blood cells in different ways.
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影响因子：
64.8
作者：
Chen, Michael J.;Yokomizo, Tomomasa;Zeigler, Brandon M.;Dzierzak, Elaine;Speck, Nancy A.
通讯作者：
Speck, Nancy A.
DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
64.5
作者：
Adolfsson, J;Månsson, R;Jacobsen, SEW
通讯作者：
Jacobsen, SEW
影响因子：
5.2
作者：
Cherqui, S;Kurian, SM;Salomon, DR
通讯作者：
Salomon, DR
影响因子：
14.9
作者：
Edgar, R;Domrachev, M;Lash, AE
通讯作者：
Lash, AE