HEATR3 variants impair nuclear import of uL18 (RPL5) and drive Diamond-Blackfan anemia
O'Donohue, Marie-Françoise; Da Costa, Lydie M; Lezzerini, Marco; Unal, Sule; Joret, Clément; Bartels, Marije; Brilstra, Eva; Scheijde-Vermeulen, Marijn; Wacheul, Ludivine; De Keersmaecker, Kim; Vereecke, Stijn; Labarque, Veerle; Saby, Manon Juliette; LeFevre, Sophie D; Platon, Jessica; Montel-Lehry, Nathalie; Laugero, Nathalie; Lacazette, Eric; van Gassen, Koen; Houtkooper, Riekelt H; Simsek-Kiper, Pelin Ozlem; Leblanc, Thierry M; Yarali, Nese; Cetinkaya, Arda; Akarsu, Nurten A; Gleizes, Pierre-Emmanuel; Lafontaine, Denis L J; MacInnes, Alyson W
(2022) Blood, volume 139, issue 21, pp. 3111 - 3126
(Article)
Abstract
The congenital bone marrow failure syndrome Diamond-Blackfan anemia (DBA) is typically associated with variants in ribosomal protein (RP) genes impairing erythroid cell development. Here we report multiple individuals with biallelic HEATR3 variants exhibiting bone marrow failure, short stature, facial and acromelic dysmorphic features, and intellectual disability. These variants destabilize a
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protein whose yeast homolog is known to synchronize the nuclear import of RPs uL5 (RPL11) and uL18 (RPL5), which are both critical for producing ribosomal subunits and for stabilizing the p53 tumor suppressor when ribosome biogenesis is compromised. Expression of HEATR3 variants or repression of HEATR3 expression in primary cells, cell lines of various origins, and yeast models impairs growth, differentiation, pre-ribosomal RNA processing, and ribosomal subunit formation reminiscent of DBA models of large subunit RP gene variants. Consistent with a role of HEATR3 in RP import, HEATR3-depleted cells or patient-derived fibroblasts display reduced nuclear accumulation of uL18. Hematopoietic progenitor cells expressing HEATR3 variants or small-hairpin RNAs knocking down HEATR3 synthesis reveal abnormal acceleration of erythrocyte maturation coupled to severe proliferation defects that are independent of p53 activation. Our study uncovers a new pathophysiological mechanism leading to DBA driven by biallelic HEATR3 variants and the destabilization of a nuclear import protein important for ribosome biogenesis.
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Keywords: Hematology, Biochemistry, Cell Biology, Immunology, Journal Article
ISSN: 0006-4971
Publisher: American Society of Hematology
Note: Funding Information: The authors offer their most special thanks to the affected individuals, their families, and to the DBA patient associations. They thank all the people involved in the mutational screening analysis in France (Christine Bole and the Team from the Genomics Core Facility, Institut Imagine-Structure Fédérative de Recherche Necker, and technicians from the Hematology Laboratory in R. Debré Hospital, Paris), Turkey (Z. Ekim Taskiran for facilitating WES studies in Hacettepe University Exome Facility and Can Kosukcu for obtaining WES data), and to the registry data manager Isabelle Marie in France. They also thank Dirk Lebrecht and Marco Teller at the University Medical Center Freiburg for the generation of the LCLs and Patricia Veltman and Petra Mooyer at the Laboratory of Genetic Metabolic Disease at the Amsterdam University Medical Center for the generation of the fibroblasts. The authors are grateful for the expert assistance of Alexia Zakaroff and Elodie Riant (Genotoul-TRI, I2MC, Toulouse) to perform cell sorting and Frédéric Martins and Emeline Lhuillier (Genotoul-GET Santé, I2MC, Toulouse) for droplet digital polymerase chain reaction. Specific E-Rare grants funding EuroDBA researchers are as follows: ZonMW #40-44000-98-1008 in the Netherlands (A.W.M.), TUBITAK 315S192 in Turkey (S.U.), and ANR-15-RAR3-0007-04 (P.-E.G. M.-F.O. and L.D.C.) in France. In addition, this project has received funding from the European Union's Horizon 2020 research and innovation programme under the EJP RD COFUND-EJP No. 825575 to P.-E.G. L.D.C. D.L.J.L. R.H.H. and A.C. Specific grants under the frame of EJP RD programme JTC 2019 RiboEurope project are as follows: TUBITAK 319S062 in Turkey (A.C.), (EJP RD/JTC2019/PINT-MULTI) grant R.8015.19, and PDR grant T.0144.20 in Belgium (D.L.J.L.), as well as ANR-19-RAR4-0016 (P.-E.G.) and ANR-19-RAR4-0016-02 (L.D.C.) in France. P.-E.G. M.-F.O. and L.D.C. were also funded by ANR-2015-AAP générique-CE12 (DBA-Multigenes). L.D.C. is additionally supported by the Laboratory of Excellence for Red Cells [(LABEX GR-Ex)-ANR Avenir-11-LABX-0005-02], and The French National PHRC OFABD (DBA registry). D.L.J.L. is supported by the Belgian Fonds de la Recherche Scientifique (F.R.S./FNRS), the Université Libre de Bruxelles (ULB), the Région Wallonne (SPW EER) (“RIBOcancer” FSO grant 1810070, POC grant 1880014), the Fonds Jean Brachet, the Internationale Brachet Stiftung, and the Epitran COST action (CA16120). K.D.K. is supported by funding from the Foundation against Cancer (P2016-081 and 2016-112), and S.V. is a fellow at FWO (1S49817N). Funding Information: Specific E-Rare grants funding EuroDBA researchers are as follows: ZonMW #40-44000-98-1008 in the Netherlands (A.W.M.), TUBITAK 315S192 in Turkey (S.U.), and ANR-15-RAR3-0007-04 (P.-E.G., M.-F.O., and L.D.C.) in France. In addition, this project has received funding from the European Union's Horizon 2020 research and innovation programme under the EJP RD COFUND-EJP No. 825575 to P.-E.G., L.D.C., D.L.J.L., R.H.H., and A.C. Specific grants under the frame of EJP RD programme JTC 2019 RiboEurope project are as follows: TUBITAK 319S062 in Turkey (A.C.), (EJP RD/JTC2019/PINT-MULTI) grant R.8015.19, and PDR grant T.0144.20 in Belgium (D.L.J.L.), as well as ANR-19-RAR4-0016 (P.-E.G.) and ANR-19-RAR4-0016-02 (L.D.C.) in France. P.-E.G., M.-F.O., and L.D.C. were also funded by ANR-2015-AAP générique-CE12 (DBA-Multigenes). L.D.C. is additionally supported by the Laboratory of Excellence for Red Cells [(LABEX GR-Ex)-ANR Avenir-11-LABX-0005-02], and The French National PHRC OFABD (DBA registry). D.L.J.L. is supported by the Belgian Fonds de la Recherche Scientifique (F.R.S./FNRS), the Université Libre de Bruxelles (ULB), the Région Wallonne (SPW EER) (“RIBOcancer” FSO grant 1810070, POC grant 1880014), the Fonds Jean Brachet, the Internationale Brachet Stiftung, and the Epitran COST action (CA16120). K.D.K. is supported by funding from the Foundation against Cancer (P2016-081 and 2016-112), and S.V. is a fellow at FWO (1S49817N). Publisher Copyright: © 2022 American Society of Hematology
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