Eltrombopag Improves Erythroid Differentiation in a Human Induced Pluripotent Stem Cell Model of Diamond Blackfan Anemia

被引:9
作者
Qanash, Husam [1 ,2 ,3 ]
Li, Yongqin [1 ]
Smith, Richard H. [1 ]
Linask, Kaari [4 ]
Young-Baird, Sara [5 ,6 ]
Hakami, Waleed [1 ,2 ,7 ]
Keyvanfar, Keyvan [8 ]
Choy, John S. [2 ]
Zou, Jizhong [4 ]
Larochelle, Andre [1 ]
机构
[1] NHLBI, Cellular & Mol Therapeut Branch, NIH, Bldg 10, Bethesda, MD 20892 USA
[2] Catholic Univ Amer, Dept Biol, Washington, DC 20064 USA
[3] Univ Hail, Dept Med Lab Sci, Coll Appl Med Sci, Hail 55476, Saudi Arabia
[4] NHLBI, IPSC Core Facil, NIH, Bldg 10, Bethesda, MD 20892 USA
[5] Eunice Kennedy Shriver Natl Inst Child Hlth & Hum, NIH, Bethesda, MD 20892 USA
[6] NIGMS, NIH, Bethesda, MD 20892 USA
[7] Jazan Univ, Dept Med Labs Technol, Coll Appl Med Sci, Jazan 45142, Saudi Arabia
[8] NHLBI, Clin Flow Core Facil, NIH, Bldg 10, Bethesda, MD 20892 USA
关键词
diamond blackfan anemia; induced pluripotent stem cells; eltrombopag; genome editing; disease modeling; drug testing;
D O I
10.3390/cells10040734
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Diamond Blackfan Anemia (DBA) is a congenital macrocytic anemia associated with ribosomal protein haploinsufficiency. Ribosomal dysfunction delays globin synthesis, resulting in excess toxic free heme in erythroid progenitors, early differentiation arrest, and pure red cell aplasia. In this study, DBA induced pluripotent stem cell (iPSC) lines were generated from blood mononuclear cells of DBA patients with inactivating mutations in RPS19 and subjected to hematopoietic differentiation to model disease phenotypes. In vitro differentiated hematopoietic cells were used to investigate whether eltrombopag, an FDA-approved mimetic of thrombopoietin with robust intracellular iron chelating properties, could rescue erythropoiesis in DBA by restricting the labile iron pool (LIP) derived from excessive free heme. DBA iPSCs exhibited RPS19 haploinsufficiency, reduction in the 40S/60S ribosomal subunit ratio and early erythroid differentiation arrest in the absence of eltrombopag, compared to control isogenic iPSCs established by CRISPR/Cas9-mediated correction of the RPS19 point mutation. Notably, differentiation of DBA iPSCs in the presence of eltrombopag markedly improved erythroid maturation. Consistent with a molecular mechanism based on intracellular iron chelation, we observed that deferasirox, a clinically licensed iron chelator able to permeate into cells, also enhanced erythropoiesis in our DBA iPSC model. In contrast, erythroid maturation did not improve substantially in DBA iPSC differentiation cultures supplemented with deferoxamine, a clinically available iron chelator that poorly accesses LIP within cellular compartments. These findings identify eltrombopag as a promising new therapeutic to improve anemia in DBA.
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页数:18
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共 61 条
[1]   Eltrombopag maintains human hematopoietic stem and progenitor cells under inflammatory conditions mediated by IFN-γ [J].
Alvarado, Luigi J. ;
Huntsman, Heather D. ;
Cheng, Hai ;
Townsley, Danielle M. ;
Winkler, Thomas ;
Feng, Xingmin ;
Dunbar, Cynthia E. ;
Young, Neal S. ;
Larochelle, Andre .
BLOOD, 2019, 133 (19) :2043-2055
[2]   A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture [J].
Beers, Jeanette ;
Linask, Kaari L. ;
Chen, Jane A. ;
Siniscalchi, Lauren I. ;
Lin, Yongshun ;
Zheng, Wei ;
Rao, Mahendra ;
Chen, Guokai .
SCIENTIFIC REPORTS, 2015, 5
[3]   TNF-mediated inflammation represses GATA1 and activates p38 MAP kinase in RPS19-deficient hematopoietic progenitors [J].
Bibikova, Elena ;
Youn, Min-Young ;
Danilova, Nadia ;
Ono-Uruga, Yukako ;
Konto-Ghiorghi, Yoan ;
Ochoa, Rachel ;
Narla, Anupama ;
Glader, Bertil ;
Lin, Shuo ;
Sakamoto, Kathleen M. .
BLOOD, 2014, 124 (25) :3791-3798
[4]   Mechanisms and therapeutic prospects of thrombopoietin receptor agonists [J].
Bussel, James ;
Kulasekararaj, Austin ;
Cooper, Nichola ;
Verma, Amit ;
Steidl, Ulrich ;
Semple, John W. ;
Will, Britta .
SEMINARS IN HEMATOLOGY, 2019, 56 (04) :262-278
[5]   Eltrombopag for the treatment of chronic idiopathic thrombocytopenic purpura [J].
Bussel, James B. ;
Cheng, Gregory ;
Saleh, Mansoor N. ;
Psaila, Bethan ;
Kovaleva, Lidia ;
Meddeb, Balkis ;
Kloczko, Janusz ;
Hassani, Habib ;
Mayer, Bhabita ;
Stone, Nicole L. ;
Arning, Michael ;
Provan, Drew ;
Jenkins, Julian M. .
NEW ENGLAND JOURNAL OF MEDICINE, 2007, 357 (22) :2237-2247
[6]  
Chen GK, 2011, NAT METHODS, V8, P424, DOI [10.1038/NMETH.1593, 10.1038/nmeth.1593]
[7]   Deferasirox improves hematologic and hepatic function with effective reduction of serum ferritin and liver iron concentration in transfusional iron overload patients with myelodysplastic syndrome or aplastic anemia [J].
Cheong, June-Won ;
Kim, Hyeoung-Joon ;
Lee, Kyoo-Hyung ;
Yoon, Sung-Soo ;
Lee, Jae Hoon ;
Park, Hee-Sook ;
Kim, Ho Young ;
Shim, Hyeok ;
Seong, Chu-Myung ;
Kim, Chul Soo ;
Chung, Jooseop ;
Hyun, Myung Soo ;
Jo, Deog-Yeon ;
Jung, Chul Won ;
Sohn, Sang Kyun ;
Yoon, Hwi-Joong ;
Kim, Byung Soo ;
Joo, Young-Don ;
Park, Chi-Young ;
Min, Yoo Hong .
TRANSFUSION, 2014, 54 (06) :1542-1551
[8]   The role of the DNA damage response in zebrafish and cellular models of Diamond Blackfan anemia [J].
Danilova, Nadia ;
Bibikova, Elena ;
Covey, Todd M. ;
Nathanson, David ;
Dimitrova, Elizabeth ;
Konto, Yoan ;
Lindgren, Anne ;
Glader, Bertil ;
Radu, Caius G. ;
Sakamoto, Kathleen M. ;
Lin, Shuo .
DISEASE MODELS & MECHANISMS, 2014, 7 (07) :895-905
[9]   Single-cell analyses demonstrate that a heme-GATA1 feedback loop regulates red cell differentiation [J].
Doty, Raymond T. ;
Yan, Xiaowei ;
Lausted, Christopher ;
Munday, Adam D. ;
Yang, Zhantao ;
Yi, Danielle ;
Jabbari, Neda ;
Liu, Li ;
Keel, Sioban B. ;
Tian, Qiang ;
Abkowitz, Janis L. .
BLOOD, 2019, 133 (05) :457-469
[10]   Drug discovery for Diamond-Blackfan anemia using reprogrammed hematopoietic progenitors [J].
Doulatov, Sergei ;
Vo, Linda T. ;
Macari, Elizabeth R. ;
Wahlster, Lara ;
Kinney, Melissa A. ;
Taylor, Alison M. ;
Barragan, Jessica ;
Gupta, Manav ;
McGrath, Katherine ;
Lee, Hsiang-Ying ;
Humphries, Jessica M. ;
DeVine, Alex ;
Narla, Anupama ;
Alter, Blanche P. ;
Beggs, Alan H. ;
Agarwal, Suneet ;
Ebert, Benjamin L. ;
Gazda, Hanna T. ;
Lodish, Harvey F. ;
Sieff, Colin A. ;
Schlaeger, Thorsten M. ;
Zon, Leonard I. ;
Daley, George Q. .
SCIENCE TRANSLATIONAL MEDICINE, 2017, 9 (376)