Experimental hematology最新文献_第3页

Emerging paradigms in redox regulation: the role of selenoproteins in normal and malignant hematopoiesis 氧化还原调控的新范式：硒蛋白在正常和恶性造血中的作用。

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-12-07 DOI: 10.1016/j.exphem.2025.105346

Yumi Aoyama , Hiromi Yamazaki , Daichi Inoue

Selenoproteins—a unique class of antioxidant enzymes characterized by the incorporation of selenocysteine at their catalytic core—function as pivotal regulators of redox homeostasis in hematopoiesis. This review elucidates how selenoprotein-mediated redox control orchestrates hematopoietic stem cell (HSC) fate decisions, maintaining the critical balance between self-renewal and differentiation. The glutathione peroxidase (GPX) family, particularly GPX1 and GPX4, plays indispensable roles in hydrogen peroxide detoxification and protection against lipid peroxidation-induced ferroptotic cell death, respectively. Similarly, thioredoxin reductases (TXNRDs) sustain critical redox equilibrium via thioredoxin regeneration. Recent studies demonstrate that these redox regulators facilitate the proliferation and differentiation of HSCs and mature lineages. Selenoprotein deficiency disrupts HSC fitness, impairs B- and erythroid-lineage maturation, and induces B-to-myeloid lineage switching—pathogenic features observed in aged hematopoiesis, highlighting the critical roles of selenoproteins in balanced, healthy hematopoiesis. Emerging evidence demonstrates that leukemic cells also exploit selenoprotein pathways to mitigate oxidative stress, suggesting that selective modulation of specific selenoproteins may constitute a promising therapeutic approach, provided we delineate their differential utilization between normal and malignant hematopoiesis. Selenoproteins function at the intersection of several transcriptional networks, including NRF2, whose orchestrated antioxidant responses may alter during aging and malignant transformation. Indeed, selenoproteins possess unique properties and function with tissue-specific expression patterns and nonredundant or redundant functions across different hematopoietic lineages. Understanding the contribution of selenoproteins to hematopoietic regulation offers promising avenues for developing targeted therapeutic strategies in hematologic disorders and rejuvenating aged hematopoiesis, potentially through precision-guided modulation of selenoprotein-dependent pathways.

硒蛋白是一类独特的抗氧化酶，其催化核心是硒半胱氨酸的结合，在造血过程中作为氧化还原稳态的关键调节因子。这篇综述阐明了硒蛋白介导的氧化还原控制如何协调造血干细胞（HSC）的命运决定，维持自我更新和分化之间的关键平衡。谷胱甘肽过氧化物酶（GPX）家族，特别是GPX1和GPX4，分别在过氧化氢解毒和保护脂质过氧化诱导的铁致细胞死亡中发挥着不可或缺的作用。类似地，硫氧还蛋白还原酶（TXNRDs）通过硫氧还蛋白再生维持临界氧化还原平衡。最近的研究表明，这些氧化还原调节剂促进造血干细胞和成熟谱系的增殖和分化。硒蛋白缺乏会破坏HSC的适应性，损害B和红系的成熟，并诱导B到髓系的转换——在老年造血中观察到的致病特征，突出了硒蛋白在平衡、健康的造血中的关键作用。新出现的证据表明，白血病细胞也利用硒蛋白途径来减轻氧化应激，这表明选择性调节特定的硒蛋白可能是一种很有前途的治疗方法，前提是我们描述了正常和恶性造血之间硒蛋白的不同利用。硒蛋白在几个转录网络的交叉点起作用，包括NRF2，其精心安排的抗氧化反应可能在衰老和恶性转化过程中发生改变。事实上，硒蛋白具有独特的性质和功能，具有组织特异性表达模式和在不同造血谱系中的非冗余或冗余功能。了解硒蛋白对造血调节的贡献，为开发针对血液系统疾病的靶向治疗策略和恢复老年造血提供了有希望的途径，可能通过精确引导硒蛋白依赖途径的调节。

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引用次数: 0

Integrin-dependence of extramedullary erythropoiesis in the spleen of Jak2-V617F positive myeloproliferative neoplasm in mice 小鼠Jak2-V617F阳性骨髓增殖性肿瘤脾脏中整合素依赖性的髓外红细胞生成。

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-12-06 DOI: 10.1016/j.exphem.2025.105340

Conny K. Baldauf , Linda Poschmann , Bärbel Edelmann-Stephan , Frank Angenstein , Tobias R. Haage , Vikas Bhuria , Lars Philipsen , Hannes Berlin , Daniela C. Dieterich , Martin Böttcher , Dimitrios Mougiakakos , Burkhart Schraven , Thomas Fischer

The molecular mechanisms driving splenomegaly in myeloproliferative neoplasms (MPNs) remain poorly understood. Utilizing the Jak2-V617F knock-in mouse model, we investigated the role of β1- and β2-integrins in regulating spleen volume and spleen weight. The response to neutralizing antibodies against VLA-4 and the β2-integrin chain, as well as to isotype controls, was evaluated by serial intraindividual magnetic resonance imaging, by assessment of spleen weight and by analysis of the cellular composition of spleens. Short-term anti-VLA-4/β2-integrin treatment (applied on day 1 and evaluated at day 8) significantly reduced the spleen volume by 30% compared with the immunoglobulin G (IgG) control. At the cellular level, anti-integrin treatment led to a substantial 30% decrease in erythroblast counts and a 23% reduction in basophilic erythroblasts within the spleen, as compared with the isotype control. Furthermore, immunohistochemistry analysis of spleen sections revealed that CD71 (= Transferrin receptor protein 1) expression in spleen remained largely unchanged, whereas there was a clear reduction in Ter119 expression upon anti-integrin treatment. These data suggest that the substantial decrease in erythroblasts following anti-integrin treatment is a primary factor contributing to the overall reduction in spleen size. To study the spleen architecture, multiepitope ligand cartography (MELC) analysis of spleen sections was applied. This demonstrated that the spatial distribution of the marginal zone, red pulp, and white pulp remained unaltered upon anti-integrin treatment in JAK2-V617F knock-in mice. In summary, the present study identified a previously unrecognized role of the β1-integrin VLA-4 and of β2-integrin chains in extramedullary erythropoiesis of the spleen in JAK2-V617F-induced disease.

骨髓增生性肿瘤（MPN）脾肿大的分子机制尚不清楚。利用Jak2-V617F敲入小鼠模型，我们研究了β1和β2整合素在调节脾脏体积和脾脏重量中的作用。通过连续的个体内MRI、脾脏重量评估和脾脏细胞组成分析来评估抗VLA-4和β2整合素链中和抗体以及同型对照的反应。短期抗vla4 /β2整合素治疗（第1天应用，第8天评估）与IgG对照组相比，脾脏体积显著减少30%。在细胞水平上，与同型对照相比，抗整合素治疗导致脾脏内红细胞计数显著减少30%，嗜碱性红细胞减少23%。此外，脾脏切片的免疫组化分析显示，CD71（=转铁蛋白受体蛋白1）在脾脏中的表达基本保持不变，而抗整合素治疗后Ter119的表达明显降低。这一数据表明，抗整合素治疗后红细胞的大量减少是导致脾脏大小总体缩小的主要因素。为了研究脾脏的结构，应用了脾脏切片的多表位配体图谱（MELC）分析。这表明JAK2-V617F敲入小鼠的边缘区、红髓和白髓的空间分布在抗整合素处理后保持不变。总之，本研究确定了β1-整合素VLA-4和β2-整合素链在jak2 - v617f诱导的疾病中脾髓外红细胞生成中先前未被认识到的作用。

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引用次数: 0

Tumor necrosis factor from leukemic environment stimulates hematopoietic stem/progenitor cells toward megakaryocyte/myeloid lineage bias 来自白血病环境的肿瘤坏死因子刺激造血干细胞/祖细胞向巨核细胞/髓细胞谱系倾斜。

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-12-04 DOI: 10.1016/j.exphem.2025.105332

Hidekazu Nishikii , Riko Kikuchi , Takaharu Kimura , Mizuki Saito , Yusuke Kiyoki , Saki Tanaka , Yamato Sasaki , Takayasu Kato , Tatsuhiro Sakamoto , Mamiko Sakata-Yanagimoto , Naoshi Obara , Satoshi Yamazaki , Shigeru Chiba

Acute myeloid leukemia (AML) is characterized by the proliferation of malignant myeloid progenitor cells and impairment of hematopoiesis. Although genetic abnormalities within leukemic cells have been investigated in detail, definitive explanations for the damage to the normal hematopoietic system are lacking. Here, we investigated the mechanisms underlying the impairment of the residual hematopoietic system in the bone marrow in AML. We evaluated the function of residual nonleukemic (nl)-hematopoietic stem/progenitor cells (HSPCs) from the bone marrow of mice with MLL-AF9-induced AML. The nl-HSPCs in the leukemic marrow showed a megakaryocyte (MgK) and myeloid-biased gene expression signature, with enrichment of tumor necrosis factor (TNF) signaling and reduced repopulation ability. To investigate whether the upregulation of TNF signaling causes the MgK/myeloid lineage bias, we investigated the effects of TNF-α in normal hematopoietic stem cells (HSCs)/HSPCs under ex vivo expansion condition. Single-cell transcriptome analysis of these cells revealed an increased frequency of cells expressing genes related to the MgK lineage and decreased repopulation capacity compared with those of ex vivo expanded HSCs/HSPCs without TNF-α. Our data suggest that increased TNF-α in the leukemic bone marrow environment at least in part drives HSPCs toward MgK/myeloid differentiation, resulting in the exhaustion of residual normal HSCs/HSPCs. These findings offer valuable insights into leukemic biology and normal hematopoiesis.

急性髓系白血病（AML）的特点是恶性髓系祖细胞增殖和造血功能受损。虽然对白血病细胞内的遗传异常进行了详细的研究，但对正常造血系统的损害还缺乏明确的解释。在这里，我们研究了AML患者骨髓中残余造血系统受损的机制。我们评估了mll - af9诱导的AML小鼠骨髓中残留的非白血病(nl)-造血干细胞/祖细胞（HSPC）的功能。白血病骨髓中的nl-HSPC表现出巨核细胞（MgK）和骨髓偏倚基因表达特征，TNF信号富集，再生能力降低。为了研究TNF信号的上调是否会导致MgK /髓系偏倚，我们在离体扩增条件下研究了TNF-α对正常造血干细胞(HSC) / HSPC的影响。这些细胞的单细胞转录组分析显示，与没有TNF-α的体外扩增HSC / HSPC相比，表达MgK谱系相关基因的细胞频率增加，再生能力下降。我们的数据表明，白血病骨髓环境中TNF-α的增加至少在一定程度上推动了HSPC向MgK /髓样分化，导致剩余的正常HSC / HSPC耗尽。这些发现为白血病生物学和正常造血提供了有价值的见解。

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引用次数: 0

Pig and human adult hematopoietic stem and progenitor cells are overall transcriptionally similar 猪和人成人造血干细胞和祖细胞在转录上总体上是相似的。

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-12-01 DOI: 10.1016/j.exphem.2025.105334

Emma Bailey , Foteini Kalampalika , Raúl Sánchez-Lanzas , Justin Barclay , Amanda Jiménez-Pompa , Jun Wang , Miguel Ganuza

Over the recent years, pigs have re-emerged as an alternative source of organs for xenotransplantation into humans with the promise to overcome a worldwide shortage of human donors. Xenotransplantation still faces critical issues with immune rejection that could be potentially solved by the generation of lymphohematopoietic chimeras. Moreover, pig hematopoietic stem and progenitor cells (HSPCs) can constitute an unlimited source of HSPCs for lifesaving HSPC transplantation in bone marrow (BM) failure and after chemotherapy, among other cell therapies. The generation of these hematopoietic chimeras requires a profound study of pig hematopoiesis including pig HSPCs. Importantly, through single-cell RNA sequencing of pig BM cells we identified pig HSPC populations transcriptionally similar to those in humans and many common transcriptional regulators of hematopoiesis evolutionarily preserved in erythromyeloid and lymphoid differentiation. This supports that hematopoiesis in pigs is hierarchically organized and regulated in a very similar fashion as in humans. We also provided a sorting strategy for the identification and isolation of several putative pig HSPC populations, which should open a new means to functionally study pig hematopoiesis.

近年来，猪再次成为异种人体器官移植的替代来源，有望克服全球范围内人类供体短缺的问题。异种移植仍然面临着免疫排斥的关键问题，这可能通过产生淋巴造血嵌合体来解决。此外，猪造血干细胞和祖细胞（HSPCs）可以作为造血干细胞的无限来源，用于骨髓衰竭和化疗后的造血干细胞移植，以及其他细胞疗法。这些造血嵌合体的产生需要对猪造血包括猪造血干细胞和祖细胞（HSPCs）进行深入的研究。重要的是，通过对猪骨髓细胞的单细胞RNA测序，我们发现猪HSPC群体在转录上与人类相似，并且在红细胞和淋巴细胞分化中进化地保留了许多常见的造血转录调节因子。这支持了猪的造血系统以与人类非常相似的方式分层组织和调节。我们还提供了一种分类策略，用于鉴定和分离几个假定的猪HSPC群体，这将为功能性研究猪造血开辟新的手段。

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引用次数: 0

Germline heterozygous SH2B3 p.Glu78Lys variant: a three-patient case series with myeloproliferative neoplasms 种系杂合SH2B3 p.g u78lys变异：骨髓增生性肿瘤（mpn）的3例病例系列

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-11-29 DOI: 10.1016/j.exphem.2025.105333

Giovanni Iaquinta , Alessandro Laganà , Anna Tamburini , Caterina Tatarelli , Patrizia Chiusolo , Elena Rossi , Monica Rossi , Michele Ragazzo , Emanuele Savino , Massimo Breccia , Paola Grammatico

We investigated the clinical significance of a rare germline SH2B3 variant (c.232G>A; p.Glu78Lys) identified by targeted next-generation sequencing (NGS) in patients with myeloproliferative neoplasms (MPNs). Among approximately 330 patients, three heterozygous carriers (≈1.0% prevalence) were identified by NGS and confirmed as germline (buccal swab) by Sanger sequencing. Two of the carriers presented with essential thrombocythemia that progressed to secondary myelofibrosis, and one presented with primary myelofibrosis that evolved to acute myeloid leukemia. The variant co-occurred with canonical somatic drivers (CALR or MPL) in the first two cases and with MPL plus additional somatic alterations (SRSF2, TET2) in the third. The p.Glu78Lys substitution localizes in the N-terminal dimerization domain of SH2B3. This germline variant is rare in population databases (allele frequency ∼1.1–2.2 per 1,000 inhabitants), and is currently classified as a variant of uncertain significance. In silico predictions were discordant, whereas structural modeling predicts disruption of critical hydrogen bonding at the dimer interface, suggesting potential functional impact. Although heterozygosity alone appears insufficient to drive disease, the enrichment of this variant in our MPN cohort and its occurrence in relatively young patients support a possible low-penetrance predisposition role. Functional assays, larger case–control series, and assessment of genetic/epigenetic modifiers are needed to define pathogenicity and clinical utility.

背景：我们研究了一种罕见的种系SH2B3变异（c.232G> a; p.Glu78Lys）在骨髓增殖性肿瘤（mpn）患者中通过靶向下一代测序（NGS）鉴定的临床意义。方法：在330例患者中，通过NGS鉴定出3例杂合携带者（患病率≈1.0%），通过Sanger测序（SS）确认为种系（口腔拭子）。结果：两名携带者表现为原发性血小板增多症（ET），发展为继发性骨髓纤维化（SMF），一名携带者表现为原发性骨髓纤维化（PMF），发展为急性髓性白血病（AML）。该变异在前两例中与典型体细胞驱动因子（CALR或MPL）共同发生，在第三例中与MPL加额外的体细胞改变（SRSF2, TET2）共同发生。p.Glu78Lys取代定位于SH2B3的n端二聚化结构域（DD）。这种种系变异在种群数据库中是罕见的（等位基因频率为每1000名居民1.1-2.2），目前被归类为不确定意义变异（VUS）。在计算机上的预测是不一致的，而结构模型预测在二聚体界面的关键氢键的破坏，表明潜在的功能影响。结论：虽然杂合性本身不足以驱动疾病，但该变异在我们的MPN队列中的富集及其在相对年轻的患者中的发生支持了可能的低外显率易感性作用。需要功能测定、更大的病例对照系列和遗传/表观遗传修饰因子的评估来确定致病性和临床应用。摘要：我们在3例骨髓增殖性肿瘤患者中报道了一种罕见的杂合种系SH2B3 （c.232G> a; p.Glu78Lys）。结构模型表明，该变体的SH2B3二聚体被破坏，这可能具有潜在的功能影响。虽然杂合性本身似乎不足以驱动疾病，但我们认为SH2B3 p.g u78lys变异可能决定了一种可能的低外显率易感性，需要功能验证和更广泛的筛选。

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引用次数: 0

ERK-mTOR crosstalk suppresses autophagy and upregulates proteasomal degradation pathway to confer chronic myeloid leukemia cells resistant to imatinib ERK-mTOR串扰抑制自噬和上调蛋白酶体降解途径，使慢性髓系白血病细胞对伊马替尼产生耐药性。

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-11-28 DOI: 10.1016/j.exphem.2025.105330

Rajdeep Roy , Tamalika Paul , Pritam Kumar Das , Samraj Sinha , Siddhartha Sankar Ray , Maitreyee Bhattacharyya , Nabendu Biswas

Drug resistance remains a critical barrier in effective cancer therapy. Previously, we demonstrated that expression of antiapoptotic protein (X‐linked inhibitor of apoptosis protein [XIAP]), contributes to the development of TRAIL resistance in chronic myeloid leukemia (CML) cells. However, upon acquiring drug resistance (K562R and KCL22R), XIAP degradation shifted from the lysosomal to the proteasomal pathway. Consistently, XIAP expression was markedly elevated in tumor samples compared with normal controls and was significantly higher in patients with an imatinib failure (IMA-FL) than in their counterparts who were imatinib responsive (IMA-RP) within the patient cohort. Moreover, we have found that proteasomal activity increased in imatinib-resistant cells and lysosomal pathway is inhibited. Mechanistically, we found that H₂O₂-induced activation of the ERK-mTOR axis suppressed autophagy in resistant cells, facilitating this shift in degradation pathway. Interestingly, dual intervention by restoring autophagic flux via mTOR inhibition and inducing XIAP degradation using H₂O₂ reverted imatinib resistance in K562R cells. Thus, our findings uncover a novel ERK–mTOR–axis for upregulation of proteasomal degradation of XIAP, which could be targeted to overcome imatinib resistance by combinatorial inhibition of mTOR and XIAP in CML. This study holds the promise of a new therapeutic strategy for overcoming drug resistance in cancer.

耐药性仍然是有效治疗癌症的关键障碍。先前，我们证实抗凋亡蛋白XIAP的表达有助于慢性髓性白血病（CML）细胞TRAIL耐药的发展。然而，在获得耐药（K562R和KCL22R）后，XIAP的降解从溶酶体途径转移到蛋白酶体途径。与正常对照相比，肿瘤样本中的XIAP表达明显升高，并且在患者队列中，伊马替尼失效（IMA-FL）患者中的XIAP表达明显高于伊马替尼应答（IMA-RP）患者。此外，我们发现伊马替尼耐药细胞的蛋白酶体活性增加，溶酶体途径受到抑制。在机制上，我们发现h2o2诱导的ERK-mTOR轴的激活抑制了抗性细胞的自噬，促进了这种降解途径的转变。非常有趣的是，通过mTOR抑制恢复自噬通量和H2O2诱导XIAP降解的双重干预可以恢复K562R细胞对伊马替尼的耐药性。因此，我们的研究结果揭示了一个新的erk -mTOR轴上调XIAP的蛋白酶体降解，可以通过联合抑制mTOR和XIAP来克服CML中的伊马替尼耐药。这项研究为克服癌症耐药性提供了一种新的治疗策略。

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引用次数: 0

Mouse Gata1 3′UTR modulates Gata1 levels to affect erythropoiesis 小鼠Gata1 3'UTR调节Gata1水平影响红细胞生成。

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-11-19 DOI: 10.1016/j.exphem.2025.105327

Ling Ling , Jiawen Huang , Zhichen Dai , Lan Yang , Fan Yang , Fangyu Gong , Xinhui Qiu , Mengying Lv , Fangfang Wang , Jingyan Liang , Sheng He , Duonan Yu

The 3′ untranslated region (3′UTR) of mRNA is crucial for post-transcriptional gene regulation, primarily through miRNAs. However, the overall role of the Gata1 3′UTR in mammals remains unclear. In this study, we knocked out the Gata1 3′UTR and observed a defect in erythropoiesis in mutant mice, evidenced by macrocytic anemia at baseline. The deletion of the Gata1 3′UTR also caused deficiencies in erythropoiesis within fetal livers. Mechanistically, removing the Gata1 3′UTR destabilizes Gata1 mRNA, leading to decreased levels of Gata1 protein. This reduced stability results from the dissociation of AU-rich elements in the 3′UTR from a trans-acting factor called ELAV-like family 1 (ELAVL1). Specifically, we conducted an RNA pulldown assay followed by mass spectrometry to identify proteins that bind to the Gata1 3′UTR. Gene Ontology analysis revealed that Elavl1 is a binding partner across nearly all categories related to mRNA stabilization. Western blotting, RNA immunoprecipitation, and mutagenesis assays confirmed the direct interaction between the Gata1 3′UTR and Elavl1. Modulating Elavl1 activity or protein levels with the small molecule inhibitor dihydro-tanshinone-I, or through ectopic expression in erythroid cells, validated Elavl1 as a stabilizing factor for Gata1 mRNA. Our results highlight the important role of the Gata1 mRNA 3′UTR in erythroid development.

mRNA的3‘非翻译区（3’ utr）对转录后基因调控至关重要，主要通过mirna进行调控。然而，Gata1 3'UTR在哺乳动物中的总体作用尚不清楚。在这项研究中，我们敲除了Gata1 3'UTR，并在突变小鼠中观察到红细胞生成缺陷，在基线时表现为大细胞性贫血。Gata1 3'UTR的缺失也会导致胎儿肝脏内红细胞生成的缺陷。从机制上讲，去除Gata1 3'UTR会破坏Gata1 mRNA的稳定性，导致Gata1蛋白水平降低。这种稳定性的降低是由于3'UTR中富含au的元素与称为ELAVL1的反式作用因子分离所致。具体来说，我们进行了RNA下拉分析，然后进行质谱分析，以鉴定与Gata1 3'UTR结合的蛋白质。基因本体分析显示，ELAVL1是几乎所有类别与mRNA稳定相关的结合伴侣。Western blotting、RNA免疫沉淀和诱变实验证实了Gata1 3'UTR和ELAVL1之间的直接相互作用。用小分子抑制剂二氢丹参酮- 1或通过红细胞中的异位表达调节ELAVL1的活性或蛋白水平，证实了ELAVL1是Gata1 mRNA的稳定因子。我们的研究结果强调了Gata1 mRNA 3'UTR在红细胞发育中的重要作用。摘要：哺乳动物Gata1 3'UTR的作用尚不清楚。在这项研究中，我们产生了Gata1 3'UTR敲除小鼠，并观察到红细胞生成缺陷，在基线时表现为大细胞性贫血。从机制上讲，去除Gata1 3'UTR会破坏Gata1 mRNA的稳定性，导致Gata1蛋白的表达降低。这种mRNA的低稳定性是由于3'UTR中富含au的元素与反式作用因子ELAVL1分离，而不是由于miRNA结合或聚A序列的缺失。我们的研究结果强调了Gata1 3'UTR在红细胞发育中的关键作用。

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引用次数: 0

IFC Editorial Board IFC编委会

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-11-19 DOI: 10.1016/S0301-472X(25)00605-8

引用次数: 0

Endothelial protein C receptor CD201 is a better marker than stem cell antigen-1 to identify mouse long-term reconstituting hematopoietic stem cells following septic challenge 内皮蛋白C受体CD201是一种比SCA1更好的标志物，用于鉴定脓毒症后小鼠长期重建造血干细胞。

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-11-19 DOI: 10.1016/j.exphem.2025.105326

Kavita Bisht, Valérie Barbier, Svetlana Shatunova, Ingrid G. Winkler, Jean-Pierre Lévesque

Stem cell antigen-1 (SCA1) is widely used to identify mouse hematopoietic stem cells (HSCs) and multipotent progenitors (MPPs) among lineage-negative KIT⁺ (LK) cells. However, SCA1 is expressed only in a few inbred mouse strains and becomes strongly upregulated in LK cells following in vivo challenge with interferons, lipopolysaccharide (LPS), or pathogens, leading to incorrect analysis of HSC functional subsets and delineation of HSC, MPP, and lineage-restricted progenitor phenotypes. Endothelial protein C receptor CD201 can be used as an alternative marker for mouse and even human HSC. However, whether CD201 expression changes following infectious challenge is unknown. Unlike SCA1, CD201 expression did not change on mouse LK cells in response to LPS in vivo. Long-term competitive transplantations with CD201⁺, CD201⁻, or SCA1⁺ LK cells showed that most reconstituting HSCs are within the LK CD201⁺ population after LPS challenge. However, the long-term competitive repopulation potential of LK SCA1⁺ cells from LPS-treated mice was much more severely reduced than that of LK CD201⁺ cells from the same LPS-treated donors, suggesting that the LK SCA1⁺ population in challenged donors becomes contaminated with CD201⁻ progenitors devoid of long-term repopulation potential. Based on the CD201 gating strategy, we reassessed the effect of LPS on HSC and MPP cycling and mobilization and their dependency on MY88 and TRIF adaptors. In conclusion, CD201 enables a more accurate analysis of mouse HSC and MPP subsets in all inbred strains in septic conditions or steady state.

干细胞抗原-1 （SCA1）被广泛用于鉴定谱系阴性KIT+ (LK)细胞中的小鼠造血干细胞（HSC）和多能祖细胞（MPP）。然而，SCA1仅在少数近交系小鼠品系中表达，在体内受到干扰素、脂多糖（LPS）或病原体的攻击后，SCA1在LK细胞中被强烈上调，导致HSC功能亚群的分析和HSC、MPP和谱系受限祖细胞亚群的描绘不正确。内皮蛋白C受体cd201可作为小鼠甚至人HSC的替代标记物。然而，CD201表达是否在感染后发生变化尚不清楚。与SCA1不同，CD201在LPS作用下在小鼠LK细胞上的表达没有变化。CD201+、CD201-或SCA1+ LK细胞的长期竞争移植表明，在LPS刺激后，大多数重组hsc都在LK CD201+群体内。然而，来自lps处理小鼠的LK SCA1+细胞的长期竞争性再生潜力比来自相同lps处理的供体的LK CD201+细胞的长期竞争性再生潜力要严重得多，这表明攻击供体中的LK SCA1+群体受到缺乏长期再生潜力的CD201祖细胞的污染。基于CD201门控策略，我们重新评估了LPS对HSC和MPP循环和动员的影响，以及它们对MY88和TRIF适配器的依赖性。总之，CD201能够更准确地分析所有近交系在脓毒症或稳态下的小鼠HSC和MPP亚群。

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引用次数: 0

Cytoreductive treatment differentially affects platelet size and cytoskeletal megakaryocyte organization during thrombopoiesis in myeloproliferative neoplasms 在骨髓增殖性肿瘤的血小板形成过程中，细胞减少治疗对血小板大小和细胞骨架巨核细胞组织有不同的影响。

IF 2.1 4区医学 Q2 HEMATOLOGY

Experimental hematology

Pub Date : 2025-11-07 DOI: 10.1016/j.exphem.2025.105288

Adela S. Cellucci , Danila B. Yañuk , Paola R. Lev , Ana C. Glembotsky , Nora P. Goette , María C. Lira , Geraldine De Luca , Laureano J. Kamiya , Paula G. Heller , Rosana F. Marta

Cytoreductive treatment is a main strategy to reduce thrombotic complications and ameliorate symptoms in Phi-negative myeloproliferative neoplasms (MPNs) comprising essential thrombocythemia, polycythemia vera, and primary myelofibrosis. Based on the observation of differences in platelet size during microscopic analysis of blood smears from MPN patients, in this work we studied whether these differences could be dependent on the type of cytoreductive drug used for patients’ treatment and whether changes in platelet size could be induced by the effect of these drugs on thrombopoiesis. Maximum platelet diameter (MPD) was measured in 120 patients with MPN. The effect of drugs on thrombopoiesis was evaluated in normal megakaryocytes (MKs) obtained from cord blood–derived CD34+ hematopoietic progenitors. Anagrelide (ANA), α-interferon (IFN), and ruxolitinib (Ruxo) increased, whereas hydroxyurea (HU) decreased platelet size. MK incubation with these drugs revealed that ANA and IFN induced abnormal proplatelet (PP) architecture and affected microtubular structure, but only ANA altered actin organization, whereas neither Ruxo nor HU modified MK cytoskeleton. By bioinformatic analysis, RANTES downregulation was identified as a candidate responsible for ANA-induced abnormalities. RANTES downregulation was confirmed in MK incubated with ANA but not with IFN. Addition of recombinant RANTES reverted ANA-induced cytoskeletal abnormalities. Evaluation of RANTES plasmatic levels and platelet RNA expression in patients with MPN showed RANTES decrease in both samples during ANA treatment, suggesting that in vitro findings could reflect ANA action in vivo. In conclusion, this study demonstrates the influence of cytoreductive drugs on platelet size and reveals their differential mechanisms of action during platelet production.

细胞减少治疗是减少血小板并发症和改善ph阴性骨髓增殖性肿瘤（mpn）症状的主要策略，包括原发性血小板增多症、真性红细胞增多症和原发性骨髓纤维化。基于我们在对MPN患者血液涂片的常规显微镜分析中观察到血小板大小的差异，在这项工作中，我们研究了这些差异是否取决于用于患者治疗的细胞减少药物的类型，以及血小板大小的变化是否可能由这些药物对血小板生成的影响引起。测量120例MPN患者的最大血小板直径。在脐带血来源的CD34+造血祖细胞中获得的正常巨核细胞（MK）中评估药物对血小板生成的影响。阿纳格列特（ANA）、α-干扰素（IFN）、鲁索利替尼（ruxolitinib）的浓度升高，羟基脲（HU）降低血小板大小。与这些药物孵育MK发现，ANA和IFN诱导异常的前血小板（PP）结构和影响微管结构，但只有ANA改变肌动蛋白组织，而Ruxo和HU没有改变MK细胞骨架。通过生物信息学分析，RANTES下调被确定为ana诱导异常的候选原因。与ANA孵育的MK证实RANTES下调，但与IFN孵育的MK未证实RANTES下调。添加重组RANTES可逆转ana诱导的细胞骨架异常。对MPN患者血浆RANTES水平和血小板RNA表达的评估显示，在ANA治疗期间，两种样本的RANTES均有所下降，表明体外研究结果可以反映ANA在体内的作用。总之，本研究证明了细胞减少药物对血小板大小的影响，并揭示了它们在血小板产生过程中的不同作用机制。结论：骨髓增殖性患者血小板大小在阿纳格列特、α-干扰素和鲁索利替尼组增加，在羟脲组减少。正常成熟巨核细胞与阿纳格列特和α-干扰素孵育，而不与鲁索利替尼和羟基脲孵育，改变前血小板结构。阿纳格列特和α-干扰素诱导微管破坏，但只有阿纳格列特改变肌动蛋白细胞骨架，降低巨核细胞RANTES的表达和释放。anagrelide诱导的异常可以通过RANTES的加入而恢复。与未治疗的患者相比，阿纳格列特治疗组RANTES血浆水平和血小板RNA表达降低，表明体外研究结果可以反映阿纳格列特在体内的作用。

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