Ribosomal and Hematopoietic Defects in Induced Pluripotent Stem Cells Derived from Diamond Blackfan Anemia Patients

Overview

Journal Blood

Publisher Elsevier

Specialty Hematology

Date 2013 Jun 8

PMID 23744582

Citations 52

Authors

Loic Garcon

Jingping Ge

Shwetha H Manjunath

Jason A Mills

Marisa Apicella

Shefali Parikh

Lisa M Sullivan

Gregory M Podsakoff

Paul Gadue

Deborah L French

Philip J Mason

Monica Bessler

Mitchell J Weiss

Affiliations

Soon will be listed here.

Abstract

Diamond Blackfan anemia (DBA) is a congenital disorder with erythroid (Ery) hypoplasia and tissue morphogenic abnormalities. Most DBA cases are caused by heterozygous null mutations in genes encoding ribosomal proteins. Understanding how haploinsufficiency of these ubiquitous proteins causes DBA is hampered by limited availability of tissues from affected patients. We generated induced pluripotent stem cells (iPSCs) from fibroblasts of DBA patients carrying mutations in RPS19 and RPL5. Compared with controls, DBA fibroblasts formed iPSCs inefficiently, although we obtained 1 stable clone from each fibroblast line. RPS19-mutated iPSCs exhibited defects in 40S (small) ribosomal subunit assembly and production of 18S ribosomal RNA (rRNA). Upon induced differentiation, the mutant clone exhibited globally impaired hematopoiesis, with the Ery lineage affected most profoundly. RPL5-mutated iPSCs exhibited defective 60S (large) ribosomal subunit assembly, accumulation of 12S pre-rRNA, and impaired erythropoiesis. In both mutant iPSC lines, genetic correction of ribosomal protein deficiency via complementary DNA transfer into the "safe harbor" AAVS1 locus alleviated abnormalities in ribosome biogenesis and hematopoiesis. Our studies show that pathological features of DBA are recapitulated by iPSCs, provide a renewable source of cells to model various tissue defects, and demonstrate proof of principle for genetic correction strategies in patient stem cells.

Citing Articles

Modeling primitive and definitive erythropoiesis with induced pluripotent stem cells.

Pavani G, Klein J, Nations C, Sussman J, Tan K, An H Blood Adv. 2024; 8(6):1449-1463.

PMID: 38290102 PMC: 10955655. DOI: 10.1182/bloodadvances.2023011708.

Establishment and characterization of CSCRi006-A: an induced pluripotent stem cell line generated from a patient with Diamond-Blackfan Anemia (DBA) carrying ribosomal protein S19 (RPS19) mutation.

Rani S, Thamodaran V, Nandy K, Fouzia N, Maddali M, Rajesh P Hum Cell. 2023; 36(6):2204-2213.

PMID: 37603219 DOI: 10.1007/s13577-023-00946-y.

De Novo Generation of Human Hematopoietic Stem Cells from Pluripotent Stem Cells for Cellular Therapy.

Ding J, Li Y, Larochelle A Cells. 2023; 12(2).

PMID: 36672255 PMC: 9857267. DOI: 10.3390/cells12020321.

Deficiency of ribosomal protein S26, which is mutated in a subset of patients with Diamond Blackfan anemia, impairs erythroid differentiation.

Piantanida N, La Vecchia M, Sculco M, Talmon M, Palattella G, Kurita R Front Genet. 2022; 13:1045236.

PMID: 36579335 PMC: 9790993. DOI: 10.3389/fgene.2022.1045236.

An RPS19-edited model for Diamond-Blackfan anemia reveals TP53-dependent impairment of hematopoietic stem cell activity.

Bhoopalan S, Yen J, Mayuranathan T, Mayberry K, Yao Y, Lillo Osuna M JCI Insight. 2022; 8(1).

PMID: 36413407 PMC: 9870085. DOI: 10.1172/jci.insight.161810.

References

Ball S . Diamond Blackfan anemia. Hematology Am Soc Hematol Educ Program. 2011; 2011:487-91. DOI: 10.1182/asheducation-2011.1.487. View

Jaako P, Flygare J, Olsson K, Quere R, Ehinger M, Henson A . Mice with ribosomal protein S19 deficiency develop bone marrow failure and symptoms like patients with Diamond-Blackfan anemia. Blood. 2011; 118(23):6087-96. DOI: 10.1182/blood-2011-08-371963. View

Matsson H, J Davey E, Draptchinskaia N, Hamaguchi I, Ooka A, Leveen P . Targeted disruption of the ribosomal protein S19 gene is lethal prior to implantation. Mol Cell Biol. 2004; 24(9):4032-7. PMC: 387766. DOI: 10.1128/MCB.24.9.4032-4037.2004. View

Quigley J, Gazda H, Yang Z, Ball S, Sieff C, Abkowitz J . Investigation of a putative role for FLVCR, a cytoplasmic heme exporter, in Diamond-Blackfan anemia. Blood Cells Mol Dis. 2005; 35(2):189-92. DOI: 10.1016/j.bcmd.2005.01.005. View

Fallon A, Jinks C, Yamamoto M, Nomura M . Expression of ribosomal protein genes cloned in a hybrid plasmid in Escherichia coli: gene dosage effects on synthesis of ribosomal proteins and ribosomal protein messenger ribonucleic acid. J Bacteriol. 1979; 138(2):383-96. PMC: 218189. DOI: 10.1128/jb.138.2.383-396.1979. View

Cretien A, Hurtaud C, Moniz H, Proust A, Marie I, Wagner-Ballon O . Study of the effects of proteasome inhibitors on ribosomal protein S19 (RPS19) mutants, identified in patients with Diamond-Blackfan anemia. Haematologica. 2008; 93(11):1627-34. DOI: 10.3324/haematol.13023. View

Angelini M, Cannata S, Mercaldo V, Gibello L, Santoro C, Dianzani I . Missense mutations associated with Diamond-Blackfan anemia affect the assembly of ribosomal protein S19 into the ribosome. Hum Mol Genet. 2007; 16(14):1720-7. DOI: 10.1093/hmg/ddm120. View

Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K . Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007; 131(5):861-72. DOI: 10.1016/j.cell.2007.11.019. View

Devlin E, Dacosta L, Mohandas N, Elliott G, Bodine D . A transgenic mouse model demonstrates a dominant negative effect of a point mutation in the RPS19 gene associated with Diamond-Blackfan anemia. Blood. 2010; 116(15):2826-35. PMC: 2974590. DOI: 10.1182/blood-2010-03-275776. View

10.

Maicas E, Pluthero F, Friesen J . The accumulation of three yeast ribosomal proteins under conditions of excess mRNA is determined primarily by fast protein decay. Mol Cell Biol. 1988; 8(1):169-75. PMC: 363097. DOI: 10.1128/mcb.8.1.169-175.1988. View

11.

Smith J, Maguire S, Davis L, Alexander M, Yang F, Chandran S . Robust, persistent transgene expression in human embryonic stem cells is achieved with AAVS1-targeted integration. Stem Cells. 2007; 26(2):496-504. DOI: 10.1634/stemcells.2007-0039. View

12.

Dutt S, Narla A, Lin K, Mullally A, Abayasekara N, Megerdichian C . Haploinsufficiency for ribosomal protein genes causes selective activation of p53 in human erythroid progenitor cells. Blood. 2010; 117(9):2567-76. PMC: 3062351. DOI: 10.1182/blood-2010-07-295238. View

13.

Danilova N, Sakamoto K, Lin S . Ribosomal protein S19 deficiency in zebrafish leads to developmental abnormalities and defective erythropoiesis through activation of p53 protein family. Blood. 2008; 112(13):5228-37. DOI: 10.1182/blood-2008-01-132290. View

14.

Flygare J, Kiefer T, Miyake K, Utsugisawa T, Hamaguchi I, Da Costa L . Deficiency of ribosomal protein S19 in CD34+ cells generated by siRNA blocks erythroid development and mimics defects seen in Diamond-Blackfan anemia. Blood. 2005; 105(12):4627-34. DOI: 10.1182/blood-2004-08-3115. View

15.

Hamaguchi I, Flygare J, Nishiura H, Brun A, Ooka A, Kiefer T . Proliferation deficiency of multipotent hematopoietic progenitors in ribosomal protein S19 (RPS19)-deficient diamond-Blackfan anemia improves following RPS19 gene transfer. Mol Ther. 2003; 7(5 Pt 1):613-22. DOI: 10.1016/s1525-0016(03)00091-1. View

16.

Taylor A, Humphries J, White R, Murphey R, Burns C, Zon L . Hematopoietic defects in rps29 mutant zebrafish depend upon p53 activation. Exp Hematol. 2011; 40(3):228-237.e5. PMC: 3310385. DOI: 10.1016/j.exphem.2011.11.007. View

17.

Raya A, Rodriguez-Piza I, Guenechea G, Vassena R, Navarro S, Barrero M . Disease-corrected haematopoietic progenitors from Fanconi anaemia induced pluripotent stem cells. Nature. 2009; 460(7251):53-9. PMC: 2720823. DOI: 10.1038/nature08129. View

18.

Fumagalli S, Ivanenkov V, Teng T, Thomas G . Suprainduction of p53 by disruption of 40S and 60S ribosome biogenesis leads to the activation of a novel G2/M checkpoint. Genes Dev. 2012; 26(10):1028-40. PMC: 3360559. DOI: 10.1101/gad.189951.112. View

19.

Li H, Haurigot V, Doyon Y, Li T, Wong S, Bhagwat A . In vivo genome editing restores haemostasis in a mouse model of haemophilia. Nature. 2011; 475(7355):217-21. PMC: 3152293. DOI: 10.1038/nature10177. View

20.

Chatr-Aryamontri A, Angelini M, Garelli E, Tchernia G, Ramenghi U, Dianzani I . Nonsense-mediated and nonstop decay of ribosomal protein S19 mRNA in Diamond-Blackfan anemia. Hum Mutat. 2004; 24(6):526-33. DOI: 10.1002/humu.20117. View