A Zebrafish Model of 5q-Syndrome Using CRISPR/Cas9 Targeting RPS14 Reveals a p53-Independent and p53-Dependent Mechanism of Erythroid Failure

Jason Ear; Jessica Hsueh; Melinda Nguyen; QingHua Zhang; Victoria Sung; Rajesh Chopra; Kathleen M. Sakamoto; Shuo Lin

doi:10.1016/j.jgg.2016.03.007

Volume 43 Issue 5

May 2016

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Article Navigation > Journal of Genetics and Genomics > 2016 > 43(5): 307-318

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A Zebrafish Model of 5q-Syndrome Using CRISPR/Cas9 Targeting RPS14 Reveals a p53-Independent and p53-Dependent Mechanism of Erythroid Failure

doi: 10.1016/j.jgg.2016.03.007

a
Department of Molecular, Cell and Developmental Biology, University of California Los Angeles, Los Angeles, CA 90095, USA
b
Celgene Corporation, San Francisco 94158, USA
c
Celgene Corporation, Summit, NJ 07901, USA
d
Division of Hematology/Oncology, Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94304, USA

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Corresponding author: E-mail address: shuolin@ucla.edu (Shuo Lin)
Received Date: 2016-01-28
Accepted Date: 2016-03-06
Rev Recd Date: 2016-02-21

Available Online: 2016-04-02

Publish Date: 2016-05-20

Abstract

Abstract

5q-syndrome is a distinct form of myelodysplastic syndrome (MDS) where a deletion on chromosome 5 is the underlying cause. MDS is characterized by bone marrow failures, including macrocytic anemia. Genetic mapping and studies using various models support the notion that ribosomal protein S14 (RPS14) is the candidate gene for the erythroid failure. Targeted disruption of RPS14 causes an increase in p53 activity and p53-mediated apoptosis, similar to what is observed with other ribosomal proteins. However, due to the higher risk for cancer development in patients with ribosome deficiency, targeting the p53 pathway is not a viable treatment option. To better understand the pathology of RPS14 deficiency in 5q-deletion, we generated a zebrafish model harboring a mutation in the RPS14 gene. This model mirrors the anemic phenotype seen in 5q-syndrome. Moreover, the anemia is due to a late-stage erythropoietic defect, where the erythropoietic defect is initially p53-independent and then becomes p53-dependent. Finally, we demonstrate the versatility of this model to test various pharmacological agents, such as RAP-011, L-leucine, and dexamethasone in order to identify molecules that can reverse the anemic phenotype.
- 5q-syndrome,
- RPS14,
- Ribosomopathy,
- Myelodysplastic syndrome

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References(25)

References

[1]	Barlow, J.L., Drynan, L.F., Hewett, D.R. et al. A p53-dependent mechanism underlies macrocytic anemia in a mouse model of human 5q-syndrome Nat. Med., 16 (2010),pp. 59-66
[2]	Barlow, J.L., Drynan, L.F., Trim, N.L. et al. New insights into 5q-syndrome as a ribosomopathy Cell Cycle, 9 (2010),pp. 4286-4293
[3]	Boultwood, J. The role of haploinsufficiency of RPS14 and p53 activation in the molecular pathogenesis of the 5q-syndrome Pediatr. Rep., 3 (2011),p. e10
[4]	Boultwood, J., Fidler, C., Lewis, S. et al. Molecular mapping of uncharacteristically small 5q deletions in two patients with the 5q-syndrome: delineation of the critical region on 5q and identification of a 5q-breakpoint Genomics, 19 (1994),pp. 425-432
[5]	Boultwood, J., Fidler, C., Strickson, A.J. et al. Narrowing and genomic annotation of the commonly deleted region of the 5q-syndrome Blood, 99 (2002),pp. 4638-4641
[6]	Boultwood, J., Pellagatti, A., Cattan, H. et al. Br. J. Haematol., 139 (2007),pp. 578-589
[7]	Chang, N., Sun, C., Gao, L. et al. Genome editing with RNA-guided Cas9 nuclease in zebrafish embryos Cell Res., 23 (2013),pp. 465-472
[8]	Danilova, N., Sakamoto, K.M., Lin, S. Ribosomal protein S19 deficiency in zebrafish leads to developmental abnormalities and defective erythropoiesis through activation of p53 protein family Blood, 112 (2008),pp. 5228-5237
[9]	, Kieran, M.W., Chan, F.Y., Barone, L.M. et al. Intraembryonic hematopoietic cell migration during vertebrate development Proc. Natl. Acad. Sci. USA, 92 (1995),pp. 10713-10717
[10]	Draptchinskaia, N., Gustavsson, P., Andersson, B. et al. The gene encoding ribosomal protein S19 is mutated in Diamond-Blackfan anaemia Nat. Genet., 21 (1999),pp. 169-175
[11]	Ear, J., Huang, H., Wilson, T. et al. RAP-011 improves erythropoiesis in zebrafish model of Diamond-Blackfan anemia through antagonizing lefty1 Blood, 126 (2015),pp. 880-890
[12]	Ebert, B.L., Pretz, J., Bosco, J. et al. Nature, 451 (2008),pp. 335-339
[13]	Ganapathi, K.A., Austin, K.M., Lee, C.S. et al. The human Shwachman-Diamond syndrome protein, SBDS, associates with ribosomal RNA Blood, 110 (2007),pp. 1458-1465
[14]	Ganis, J.J., Hsia, N., Trompouki, E. et al. Zebrafish globin switching occurs in two developmental stages and is controlled by the LCR Dev. Biol., 366 (2012),pp. 185-194
[15]	Jowett, T. Analysis of protein and gene expression Methods Cell Biol., 59 (1999),pp. 63-85
[16]	Kok, F.O., Shin, M., Ni, C.W. et al. Reverse genetic screening reveals poor correlation between morpholino-induced and mutant phenotypes in zebrafish Dev. Cell, 32 (2015),pp. 97-108
[17]	List, A., Dewald, G., Bennett, J. et al. Lenalidomide in the myelodysplastic syndrome with chromosome 5q deletion N. Engl. J. Med., 355 (2006),pp. 1456-1465
[18]	List, A., Kurtin, S., Roe, D.J. et al. Efficacy of lenalidomide in myelodysplastic syndromes N. Engl. J. Med., 352 (2005),pp. 549-557
[19]	Luzzatto, L., Karadimitris, A. Dyskeratosis and ribosomal rebellion Nat. Genet., 19 (1998),pp. 6-7
[20]	Miliani de Marval, P.L., Zhang, Y. The RP-Mdm2-p53 pathway and tumorigenesis Oncotarget, 2 (2011),pp. 234-238
[21]	Narla, A., Payne, E.M., Abayasekara, N. et al. L-Leucine improves the anaemia in models of Diamond Blackfan anaemia and the 5q-syndrome in a TP53-independent way Br. J. Haematol., 167 (2014),pp. 524-528
[22]	Thiel, C.T., Rauch, A. The molecular basis of the cartilage-hair hypoplasia-anauxetic dysplasia spectrum Best Pract. Res. Clin. Endocrinol. Metab., 25 (2011),pp. 131-142
[23]	Wei, S., Chen, X., McGraw, K. et al. Lenalidomide promotes p53 degradation by inhibiting MDM2 auto-ubiquitination in myelodysplastic syndrome with chromosome 5q deletion Oncogene, 32 (2013),pp. 1110-1120
[24]	Wu, L., Li, X., Xu, F. et al. Low RPS14 expression in MDS without 5q – aberration confers higher apoptosis rate of nucleated erythrocytes and predicts prolonged survival and possible response to lenalidomide in lower risk non-5q-patients Eur. J. Haematol., 90 (2013),pp. 486-493
[25]	Zhang, Y., Ear, J., Yang, Z. et al. Defects of protein production in erythroid cells revealed in a zebrafish Diamond-Blackfan anemia model for mutation in RPS19 Cell Death Dis., 5 (2014),p. e1352