Anemia in low-risk myelodysplastic neoplasms (LR-MDS) derives from defective maturation and apoptosis of erythroid precursors [1] and its first-line (1L) treatment relies on erythropoiesis stimulating agents (ESAs) in case of serum EPO < 200 U/L. Still, 30–40% of patients selected with a high probability of response are resistant [2].

低风险骨髓增生异常肿瘤（LR-MDS）中的贫血源于红细胞前体的成熟和凋亡缺陷[1]，其一线（1L）治疗依赖于血清EPO<200U/L的红细胞生成刺激剂（ESAs）。仍然有30-40%的高反应概率患者具有耐药性[2]。

The knowledge of MDS mutational landscape has paved the way for molecular classification (WHO 2022 [3], ICC [4]) and IPSS-M prognostic risk stratification [5], but their role in ESA response has not been completely elucidated.The availability of novel agents like Luspatercept, which targets late erythroid precursors, and Imetelstat has in fact prompted this need of precision in order to avoid ineffective and costly treatments [6, 7].Since ESAs act on early erythroid precursors [8], we exploited multiparametric flow cytometry (MFC) to analyze baseline erythroid subpopulations [9] from ESA treated LR-MDS patients.

MDS突变景观的知识为分子分类（WHO 2022（3），ICC（4））和IPSS-M预后风险分层（5）铺平了道路，但它们在ESA反应中的作用尚未完全阐明。靶向晚期红细胞前体的新型药物如Luspatercept和Imetelstat的可用性实际上促使了对精确度的需求，以避免无效和昂贵的治疗[6，7]。由于ESA作用于早期红细胞前体（8），我们利用多参数流式细胞术（MFC）分析了ESA治疗的LR-MDS患者的基线红细胞亚群（9）。

Results were correlated with ESA response and validated in an external cohort.Composition of erythroid subpopulations and ESA response were also correlated with both MDS classification and IPSS-M risk categories. We believe that integrating MFC and molecular information at baseline would improve the decision-making for 1L treatment in anemic LR-MDS.Bone marrow (BM) cells from LR-MDS (n = 87 from the learning cohort and n = 54 from the validation cohort) were analyzed with MFC and targeted next generation sequencing (t-NGS, n = 83 and 44, respectively) analyzed presence of somatic mutations in 35 genes before ESA treatment.

结果与ESA反应相关，并在外部队列中得到验证。红细胞亚群的组成和ESA反应也与MDS分类和IPSS-M风险类别相关。我们认为，在基线时整合MFC和分子信息将改善贫血LR-MDS中1L治疗的决策。用MFC和靶向下一代测序（t-NGS，分别为83和44）分析了来自LR-MDS的骨髓（BM）细胞（来自学习队列的n=87和来自验证队列的n=54）在ESA处理之前分析了35个基因中体细胞突变的存在。

We evaluated by MFC the repartition of erythroid (ery) early precursors: ery-HPCs/CD34+cells, ery-CD117+/CD117+ and ery-CD117+/ery (see Suppl. for gating and definitions) and correlated with ESA response. In addi.

我们通过MFC评估了红系（ery）早期前体的重新分配：ery HPC/CD34+细胞，ery-CD117+/CD117+和ery-CD117+/ery（有关门控和定义，请参见附录），并与ESA反应相关。另外。

ReferencesClaessens YE, Bouscary D, Dupont JM, Picard F, Melle J, Gisselbrecht S, et al. In vitro proliferation and differentiation of erythroid progenitors from patients with myelodysplastic syndromes: evidence for Fas-dependent apoptosis. Blood. 2002;99:1594–601.Article

参考文献Claessens YE，Boutrarray D，Dupont JM，Picard F，Melle J，Gisselbrecht S等。骨髓增生异常综合征患者红系祖细胞的体外增殖和分化：Fas依赖性细胞凋亡的证据。血。2002年；99:1594-601.文章

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Park S, Grabar S, Kelaidi C, Beyne-Rauzy O, Picard F, Bardet V, et al. Predictive factors of response and survival in myelodysplastic syndrome treated with erythropoietin and G-CSF: the GFM experience. Blood. 2008;111:574–82.Article

Park S，Grabar S，Kelaidi C，Beyne Rauzy O，Picard F，Bardet V等。促红细胞生成素和G-CSF治疗骨髓增生异常综合征的反应和生存预测因素：GFM经验。血。2008年；111:574-82.文章

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Khoury JD, Solary E, Abla O, Akkari Y, Alaggio R, Apperley JF, et al. The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours: Myeloid and Histiocytic/Dendritic Neoplasms. Leukemia. 2022;36:1703–19.Article

Khoury JD，Solary E，Abla O，Akkari Y，Alaggio R，Apperley JF等。世界卫生组织血液淋巴肿瘤分类第5版：骨髓和组织细胞/树突状肿瘤。白血病。2022年；36:1703–19.文章

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Arber DA, Orazi A, Hasserjian RP, Borowitz MJ, Calvo KR, Kvasnicka HM, et al. International Consensus Classification of Myeloid Neoplasms and Acute Leukemias: integrating morphologic, clinical, and genomic data. Blood 2022;140:1200–28.Article

Arber DA，Orazi A，Hasserjian RP，Borowitz MJ，Calvo KR，Kvasnicka HM等。骨髓肿瘤和急性白血病的国际共识分类：整合形态学，临床和基因组数据。血液2022；140:1200–28.文章

CAS

中科院

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Bernard E, Tuechler H, Greenberg PL, Hasserjian RP, Arango OJE, Nannya Y, et al. Molecular International Prognostic Scoring System for Myelodysplastic Syndromes. NEJM Evid. 2022;1:EVIDoa2200008.Article

Bernard E，Tuechler H，Greenberg PL，Hasserjian RP，Arango OJE，Nannya Y等。骨髓增生异常综合征的分子国际预后评分系统。内杰姆·埃维德。2022年；1： EVIDoa2200008。文章

PubMed

PubMed

Google Scholar

谷歌学者

Della Porta M, Platzbecker U, Santini V, Garcia-Manero G, Komrokji RS, Ito R, et al. The commands trial: A Phase 3 study of the efficacy and safety of Luspatercept Versus Epoetin Alfa for the treatment of anemia due to IPSS-R very low-, low-, or intermediate-risk MDS in Erythropoiesis stimulating agent-naive patients who require RBC transfusions.

Della Porta M，Platzbecker U，Santini V，Garcia-Manero G，Komrokji RS，Ito R等。commands试验：Luspatercept与Epoetin Alfa治疗IPSS-R引起的贫血的疗效和安全性的3期研究非常低，低或中等风险的MDS在需要红细胞输注的未使用红细胞生成刺激剂的患者中。

Blood. 2020;136:1–2.Article .

血。2020年；136:1-2。文章。

Google Scholar

谷歌学者

Platzbecker U, Santini V, Fenaux P, Sekeres MA, Savona MR, Madanat YF, et al. Imetelstat in patients with lower-risk myelodysplastic syndromes who have relapsed or are refractory to erythropoiesis-stimulating agents (IMerge): a multinational, randomised, double-blind, placebo-controlled, phase 3 trial.

Platzbecker U，Santini V，Fenaux P，Sekeres MA，Savona MR，Madanat YF等。Imetelstat治疗复发或对红细胞生成刺激剂难治的低风险骨髓增生异常综合征患者（IMerge）：一项跨国，随机，双盲，安慰剂对照的3期临床试验。

Lancet. 2024;403:249–60.Article .

柳叶刀。2024年；403:249-60。文章。

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Bhoopalan SV, Huang LJS, Weiss MJ. Erythropoietin regulation of red blood cell production: from bench to bedside and back. F1000Research; 2020 [cited 2023 Jan 14]. Available from: https://f1000research.com/articles/9-1153.Orfao A, Matarraz S, Pérez-Andrés M, Almeida J, Teodosio C, Berkowska MA, et al.

Bhoopalan SV，Huang LJS，Weiss MJ。促红细胞生成素调节红细胞生成：从长凳到床边和背部。F1000研究；2020年[引自2023年1月14日]。可从以下地址获得：https://f1000research.com/articles/9-1153.OrfaoA，Matarraz S，Pérez AndréS M，Almeida J，Teodosio C，Berkowska MA等。

Immunophenotypic dissection of normal hematopoiesis. J Immunol Methods. 2019;475:112684.Article .

正常造血的免疫表型解剖。J免疫方法。2019年；475:112684。条款。

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Platzbecker U, Fenaux P, Adès L, Giagounidis A, Santini V, van de Loosdrecht AA, et al. Proposals for revised IWG 2018 hematological response criteria in patients with MDS included in clinical trials. Blood. 2019;133:1020–30.Article

Platzbecker U，Fenaux P，Adès L，Giagounidis A，Santini V，van de Loosdrecht AA等。临床试验中MDS患者修订IWG 2018血液学反应标准的建议。血。2019年；133:1020–30.文章

CAS

中科院

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Garderet L, Kobari L, Mazurier C, Witte CD, Giarratana MC, Pérot C, et al. Unimpaired terminal erythroid differentiation and preserved enucleation capacity in myelodysplastic 5q(del) clones: a single cell study. Haematologica. 2010;95:398–405.Article

Garderet L，Kobari L，Mazurier C，Witte CD，Giarratana MC，Pérot C等。骨髓增生异常5q（del）克隆中未受损的终末红细胞分化和保留的去核能力：单细胞研究。血液学。2010年；95:398-405.文章

PubMed

PubMed

Google Scholar

谷歌学者

Raimbault A, Itzykson R, Willems L, Rousseau A, Chapuis N, Mathis S, et al. The fraction of CD117/c-KIT-expressing erythroid precursors predicts ESA response in low-risk myelodysplastic syndromes. Cytom Part B: Clin Cytom. 2019;96:215–22.Article

Raimbault A，Itzykson R，Willems L，Rousseau A，Chapuis N，Mathis S等。表达CD117/c-KIT的红细胞前体的比例可预测ESA对低风险骨髓增生异常综合征的反应。Cytom B部分：临床Cytom。2019年；96:215–22.文章

CAS

中科院

Google Scholar

谷歌学者

Lieu YK, Liu Z, Ali AM, Wei X, Penson A, Zhang J, et al. SF3B1 mutant-induced missplicing of MAP3K7 causes anemia in myelodysplastic syndromes. Proc Natl Acad Sci. 2022;119:e2111703119.Article

LiueYK，Liu Z，Ali AM，Wei X，Penson A，Zhang J等。SF3B1突变体诱导的MAP3K7错配导致骨髓增生异常综合征贫血。美国国家科学院院刊。2022年；119:e2111703119.文章

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Visconte V, Mahfouz RZ, Barnard J, Tabarroki A, Zhang L, Hasrouni E, et al. Splicing Factor 3b Subunit 1 (SF3B1) mediates Mitochondrial Iron Overload In Myelodysplastic Syndromes With Ring Sideroblasts By Alternative Splicing Of Mitoferrin-1 (SLC25A37). Blood. 2013;122:1555.Article

Visconte V，Mahfouz RZ，Barnard J，Tabarroki A，Zhang L，Hasrouni E等。剪接因子3b亚基1（SF3B1）通过选择性剪接线粒体铁蛋白-1（SLC25A37）介导具有环状铁粒幼细胞的骨髓增生异常综合征中的线粒体铁超负荷。血。2013年；122:1555.文章

Google Scholar

谷歌学者

Rombaut D, Lefèvre C, Rached T, Bondu S, Letessier A, Mangione RM, et al. Accelerated DNA replication fork speed due to loss of R-loops in myelodysplastic syndromes with SF3B1 mutation. Nat Commun. 2024;15:3016.Article

Rombaut D，Lefèvre C，Rached T，Bondu S，Letessier A，Mangione RM等。由于SF3B1突变的骨髓增生异常综合征中R环的丢失，加速了DNA复制叉的速度。纳特公社。2024年；15： 3016条

CAS

中科院

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Download referencesAcknowledgementsThe authors are grateful to the patients who consented to participate in this study. This study was partially supported by Associazione Italiana per la Ricerca sul Cancro (AIRC) IG-26537-2021 Investigator Research Grant.Author informationAuthors and AffiliationsMDS Unit, DMSC, AOU Careggi, University of Florence, Florence, ItalyMarco G.

下载参考文献致谢作者感谢同意参加本研究的患者。这项研究得到了意大利协会（Associazione Italiana per la Ricerca sul Cancro）（AIRC）IG-26537-2021研究员研究资助的部分支持。作者信息作者和附属机构DMSC，AOU Careggi，佛罗伦萨大学，佛罗伦萨，ItalyMarco G。

Raddi, Giorgio Mattiuz, Sven De Pourcq, Angela Consagra, Luca Rigodanza, Cristina Amato, Elena Tofacchi, Enrico Attardi & Valeria SantiniDepartment of Medical Biotechnologies, University of Siena, Siena, ItalyMarco G. Raddi, Sven De Pourcq & Elena TofacchiFlow Cytometry and Immunotherapy Diagnostic Center, AOU Careggi, Florence, ItalySara Bencini, Benedetta Peruzzi & Francesco AnnunziatoDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, University of Florence, Florence, ItalyMichele TanturliUniversité Paris Cité and Assistance Publique-Hôpitaux de Paris.

Raddi，Giorgio Mattiuz，Sven De Pourcq，Angela Consagra，Luca Rigodanza，Cristina Amato，Elena Tofacchi，Enrico Attardi＆Valeria Santini医学生物技术系，锡耶纳大学，锡耶纳，ItalyMarco G.Raddi，Sven De Pourcq＆Elena Tofacchi流式细胞术和免疫治疗诊断中心，AOU Careggi，佛罗伦萨，ItalySara Bencini，Benedetta Peruzzi＆Francesco AnnunziatoDepartment of Experimental and Clinical Biomedical Sciences“Mario Serio”，佛罗伦萨大学，佛罗伦萨巴黎城市图尔留大学（TurliuniversitéParis Cité）和巴黎公共图书馆（Assistance Publique-Hôpitaux De Paris）。

Hôpital Cochin, Laboratory of Hematology, Paris, FranceNicolas Chapuis, Olivier Kosmider & Michaela FontenayAOU Careggi, Hematology department, Florence, ItalyLuca Rigodanza, Alessandro Sanna & Valeria SantiniGrenoble Alpes Hospital, Hematology Department, University Grenoble Alpes, Grenoble, FranceSophie ParkDepartment of Experimental and Clinical Medicine and DENOTHE Center, University of Florence, Florence, ItalyFrancesco AnnunziatoAuthorsMarco G.

Hôpital Cochin，巴黎血液学实验室，FranceNicolas Chapuis，Olivier Kosmider＆Michaela FontenayAOU Careggi，佛罗伦萨血液学系，ItalyLuca Rigodanza，Alessandro Sanna＆Valeria Santinigrenobe Alpes医院，血液学系，格勒诺布尔阿尔卑斯大学，格勒诺布尔，弗朗西斯科·帕克实验与临床医学系和佛罗伦萨大学佛罗伦萨分校DENOTHE中心，佛罗伦萨，ItalyFrancesco Annunziatoauthousmarco G。

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PubMed Google ScholarContributionsMGR and VS designed the study, followed patients, and analyzed data. FA approved flow cytometry analysis and SB and BP performed MFC analysis of cases from the learning cohort, while MGR and NC contributed to MFC analysis of cases from the validation cohort.

PubMed Google ScholarContributionsMGR和VS设计了这项研究，跟踪患者并分析了数据。FA批准的流式细胞仪分析，SB和BP对学习队列中的病例进行了MFC分析，而MGR和NC对验证队列中的病例进行了MFC分析。

MF and SP followed patients provided data for the validation cohort and participated to manuscript writing. OK performed NGS analysis. MT and SDP performed statistical analysis. GM contributed to the design of the study, the revision of the manuscript, and to the optimization of tables and figures. EA, ET, AC, LR, CA.

MF和SP跟踪患者为验证队列提供数据，并参与手稿撰写。OK进行了NGS分析。MT和SDP进行了统计分析。GM为研究的设计，手稿的修订以及表格和图形的优化做出了贡献。EA，ET，AC，LR，CA。

and AS contributed to the interpretation of results and approved the manuscript.Corresponding authorCorrespondence to.

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Reprints and permissionsAbout this articleCite this articleRaddi, M.G., Bencini, S., Peruzzi, B. et al. Flow cytometric analysis of erythroid precursors and mutational signatures of lower risk myelodysplastic syndromes identify responders to erythroid stimulating agents.

转载和许可本文引用本文Raddi，M.G.，Bencini，S.，Peruzzi，B。等人。红细胞前体的流式细胞术分析和低风险骨髓增生异常综合征的突变特征确定了对红细胞刺激剂的反应者。

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