Deletion of Asxl1 results in myelodysplasia and severe developmental defects in vivo

Journal of Experimental Medicine

Volumn 210, Issue 12, 2013, Pages 2641-2659

  Abdel Wahab, Omar ^a   Gao, Jie ^c   Adli, Mazhar ^d   Dey, Anwesha ^g   Trimarchi, Thomas ^c   Chung, Young Rock ^a   Kuscu, Cem ^d   Hricik, Todd ^a   Ndiaye Lobry, Delphine ^c   LaFave, Lindsay M ^a,b   Koche, Richard ^e,f   Shih, Alan H ^a   Guryanova, Olga A ^a   Kim, Eunhee ^a   Li, Sheng ^h   Pandey, Suveg ^a   Shin, Joseph Y ^a   Telis, Leon ^a   Liu, Jinfeng ^g   Bhatt, Parva K ^a   Monette, Sebastien ⁱ   Zhao, Xinyang ^j   Mason, Christopher E ^h   Park, Christopher Y ^a   Bernstein, Bradley E ^e,f   Aifantis, Iannis ^c   Levine, Ross L ^a,b,h   more..

^a MEMORIAL SLOAN KETTERING CANCER CENTER   (United States)

^b Gerstner Sloan Kettering Graduate School of Biomedical Sciences   (United States)

^c NEW YORK UNIVERSITY SCHOOL OF MEDICINE   (United States)

^d UNIVERSITY OF VIRGINIA   (United States)

^e BROAD INSTITUTE OF MIT AND HARVARD   (United States)

^f MASSACHUSETTS GENERAL HOSPITAL   (United States)

^g GENENTECH INC   (United States)

^h WEILL CORNELL MEDICAL COLLEGE   (United States)

ⁱ ROCKEFELLER UNIVERSITY   (United States)

^j UNIVERSITY OF ALABAMA AT BIRMINGHAM   (United States)

more..

Author keywords

[No Author keywords available]

Indexed keywords

ADDITION OF SEX COMB LIKE 1 PROTEIN; HISTONE H3; HISTONE H3 LYSINE 27; POLYCOMB GROUP PROTEIN; UNCLASSIFIED DRUG;

ALLELE; ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ANOPHTHALMIA; ARTICLE; ASXL1 GENE; CELL DIFFERENTIATION; CELL RENEWAL; CHIMERA; CHROMATIN IMMUNOPRECIPITATION; CLEFT PALATE; CONTROLLED STUDY; DEATH; DEVELOPMENTAL DISORDER; DNA SEQUENCE; EPIGENETICS; GENE; GENE DELETION; GENE EXPRESSION; GENE LOSS; GENETIC ASSOCIATION; HEMATOPOIESIS; HEMATOPOIETIC STEM CELL; HEMATOPOIETIC STEM CELL TRANSPLANTATION; HISTONE METHYLATION; JAW MALFORMATION; KNOCKOUT MOUSE; LATENT PERIOD; LETHALITY; MICROCEPHALY; MOUSE; MYELODYSPLASTIC SYNDROME; NONHUMAN; PHENOTYPE; PRIORITY JOURNAL; PROTEIN EXPRESSION; REGULATORY MECHANISM; RNA SEQUENCE; TEN ELEVEN TRANSLOCATION 2 GENE;

ABNORMALITIES, MULTIPLE; ANIMALS; DISEASE MODELS, ANIMAL; DNA-BINDING PROTEINS; EPIGENESIS, GENETIC; FEMALE; GENE DELETION; GERM-LINE MUTATION; HEMATOPOIESIS; HEMATOPOIETIC STEM CELLS; HUMANS; MICE; MICE, 129 STRAIN; MICE, INBRED C57BL; MICE, KNOCKOUT; MUTATION; MYELODYSPLASTIC SYNDROMES; PHENOTYPE; PREGNANCY; PROTEIN BINDING; PROTO-ONCOGENE PROTEINS; REPRESSOR PROTEINS;

EID: 84888116023     PISSN: 00221007     EISSN: 15409538     Source Type: Journal    
DOI: 10.1084/jem.20131141     Document Type: Article

References (55)

1. Abdel-Wahab, O., M. Adli, L.M. LaFave, J. Gao, T. Hricik, A.H. Shih, S. Pandey, J.P. Patel, Y.R. Chung, R. Koche, et al. 2012. ASXL1 mutations promote myeloid transformation through loss of PRC2-mediated gene repression. Cancer Cell. 22, 180-193 http://dx.doi.org/10.1016/j.ccr.2012.06.032.
2. Bejar, R., K. Stevenson, O. Abdel-Wahab, N. Galili, B. Nilsson, G. Garcia-Manero, H. Kantarjian, A. Raza, R.L. Levine, D. Neuberg, and B.L. Ebert. 2011. Clinical effect of point mutations in myelodysplastic syndromes. N. Engl. J. Med. 364, 2496-2506 http://dx.doi.org/10.1056/NEJMoa1013343.
3. Bejar, R., K.E. Stevenson, B.A. Caughey, O. Abdel-Wahab, D.P. Steensma, N. Galili, A. Raza, H. Kantarjian, R.L. Levine, D. Neuberg, et al. 2012. Validation of a prognostic model and the impact of mutations in patients with lower-risk myelodysplastic syndromes. J. Clin. Oncol. 30, 3376-3382 http://dx.doi.org/10.1200/JCO.2011.40.7379.
4. Boultwood, J., J. Perry, A. Pellagatti, M. Fernandez-Mercado, C. Fernandez-Santamaria, M.J. Calasanz, M.J. Larrayoz, M. Garcia-Delgado, A. Giagounidis, L. Malcovati, et al. 2010. Frequent mutation of the polycomb-associated gene ASXL1 in the myelodysplastic syndromes and in acute myeloid leukemia. Leukemia. 24, 1062-1065 http://dx.doi.org/10.1038/leu.2010.20.
5. Bracken, A.P., D. Kleine-Kohlbrecher, N. Dietrich, D. Pasini, G. Gargiulo, C. Beekman, K. Theilgaard-Mönch, S. Minucci, B.T. Porse, J.C. Marine, et al. 2007. The Polycomb group proteins bind throughout the INK4A-ARF locus and are disassociated in senescent cells. Genes Dev. 21, 525-530 http://dx.doi.org/10.1101/gad.415507.
6. Choi, C.W., Y.J. Chung, C. Slape, and P.D. Aplan. 2008. Impaired differentiation and apoptosis of hematopoietic precursors in a mouse model of myelodysplastic syndrome. Haematologica. 93, 1394-1397 http://dx.doi.org/10.3324/haematol.13042.
7. de Pontual, L., E. Yao, P. Callier, L. Faivre, V. Drouin, S. Cariou, A. Van Haeringen, D. Geneviève, A. Goldenberg, M. Oufadem, et al. 2011. Germline deletion of the miR-17?92 cluster causes skeletal and growth defects in humans. Nat. Genet. 43, 1026-1030 http://dx.doi.org/10.1038/ng.915.
8. Dey, A., D. Seshasayee, R. Noubade, D.M. French, J. Liu, M.S. Chaurushiya, D.S. Kirkpatrick, V.C. Pham, J.R. Lill, C.E. Bakalarski, et al. 2012. Loss of the tumor suppressor BAP1 causes myeloid transformation. Science. 337, 1541-1546 http://dx.doi.org/10.1126/science.1221711.
9. Fisher, C.L., I. Lee, S. Bloyer, S. Bozza, J. Chevalier, A. Dahl, C. Bodner, C.D. Helgason, J.L. Hess, R.K. Humphries, and H.W. Brock. 2010a. Additional sex combs-like 1 belongs to the enhancer of trithorax and polycomb group and genetically interacts with Cbx2 in mice. Dev. Biol. 337, 9-15 http://dx.doi.org/10.1016/j.ydbio.2009.10.004.
10. Fisher, C.L., N. Pineault, C. Brookes, C.D. Helgason, H. Ohta, C. Bodner, J.L. Hess, R.K. Humphries, and H.W. Brock. 2010b. Loss-of-function Additional sex combs like 1 mutations disrupt hematopoiesis but do not cause severe myelodysplasia or leukemia. Blood. 115, 38-46 http://dx.doi .org/10.1182/blood-2009-07-230698.
11. Gelsi-Boyer, V., V. Trouplin, J. Adélaïde, J. Bonansea, N. Cervera, N. Carbuccia, A. Lagarde, T. Prebet, M. Nezri, D. Sainty, et al. 2009. Mutations of polycomb-associated gene ASXL1 in myelodysplastic syndromes and chronic myelomonocytic leukaemia. Br. J. Haematol. 145, 788-800 http://dx.doi.org/10.1111/j.1365-2141.2009.07697.x.
12. Gilliland, D.G. 2001. The diverse role of the ETS family of transcription factors in cance. Clin. Cancer Res. 7:451-453.
13. Haferlach, C., U. Bacher, S. Schnittger, T. Alpermann, M. Zenger, W. Kern, and T. Haferlach. 2012. ETV6 rearrangements are recurrent in myeloid malignancies and are frequently associated with other genetic events. Genes Chromosomes Cancer. 51, 328-337 http://dx.doi.org/10.1002/gcc.21918.
14. Hidalgo, I., A. Herrera-Merchan, J.M. Ligos, L. Carramolino, J. Nuñez, F. Martinez, O. Dominguez, M. Torres, and S. Gonzalez. 2012. Ezh1 is required for hematopoietic stem cell maintenance and prevents senescencelike cell cycle arrest. Cell Stem Cell. 11, 649-662 http://dx.doi.org/10 .1016/j.stem.2012.08.001.
15. Hoischen, A., B.W. van Bon, B. Rodríguez-Santiago, C. Gilissen, L.E. Vissers, P. de Vries, I. Janssen, B. van Lier, R. Hastings, S.F. Smithson, et al. 2011. De novo nonsense mutations in ASXL1 cause Bohring-Opitz syndrome. Nat. Genet. 43, 729-731 http://dx.doi.org/10.1038/ng.868.
16. Itzykson, R., O. Kosmider, A. Renneville, V. Gelsi-Boyer, M. Meggendorfer, M. Morabito, C. Berthon, L. Adès, P. Fenaux, O. Beyne-Rauzy, et al. 2013. Prognostic score including gene mutations in chronic myelomonocytic leukemia. J. Clin. Oncol. 31, 2428-2436 http://dx.doi.org/10 .1200/JCO.2012.47.3314.
17. Ivanova, N.B., J.T. Dimos, C. Schaniel, J.A. Hackney, K.A. Moore, and I.R. Lemischka. 2002. A stem cell molecular signature. Science. 298, 601-604 http://dx.doi.org/10.1126/science.1073823.
18. Jacobs, J.J., K. Kieboom, S. Marino, R.A. DePinho, and M. van Lohuizen. 1999. The oncogene and Polycomb-group gene bmi-1 regulates cell proliferation and senescence through the ink4a locus. Nature. 397, 164-168 http://dx.doi.org/10.1038/16476.
19. Jankowska, A.M., H. Makishima, R.V. Tiu, H. Szpurka, Y. Huang, F. Traina, V. Visconte, Y. Sugimoto, C. Prince, C. O'Keefe, et al. 2011. Mutational spectrum analysis of chronic myelomonocytic leukemia includes genes associated with epigenetic regulation: UTX, EZH2, and DNMT3A. Blood. 118, 3932-3941 http://dx.doi.org/10.1182/blood-2010-10-311019.
20. Klinakis, A., C. Lobry, O. Abdel-Wahab, P. Oh, H. Haeno, S. Buonamici, I. van De Walle, S. Cathelin, T. Trimarchi, E. Araldi, et al. 2011. A novel tumour-suppressor function for the Notch pathway in myeloid leukaemia. Nature. 473, 230-233 http://dx.doi.org/10.1038/nature09999.
21. Ko, M., H.S. Bandukwala, J. An, E.D. Lamperti, E.C. Thompson, R. Hastie, A. Tsangaratou, K. Rajewsky, S.B. Koralov, and A. Rao. 2011. Ten-Eleven-Translocation 2 (TET2) negatively regulates homeostasis and differentiation of hematopoietic stem cells in mice. Proc. Natl. Acad. Sci. USA. 108, 14566-14571 http://dx.doi.org/10.1073/pnas.1112317108.
22. Koschmieder, S., F. Rosenbauer, U. Steidl, B.M. Owens, and D.G. Tenen. 2005. Role of transcription factors C/EBPalpha and PU.1 in normal hematopoiesis and leukemia. Int. J. Hematol. 81, 368-377 http://dx.doi.org/10.1532/IJH97.05051.
23. Kühn, R., F. Schwenk, M. Aguet, and K. Rajewsky. 1995. Inducible gene targeting in mice. Science. 269, 1427-1429 http://dx.doi.org/10.1126/science.7660125.
24. Lakso, M., J.G. Pichel, J.R. Gorman, B. Sauer, Y. Okamoto, E. Lee, F.W. Alt, and H. Westphal. 1996. Efficient in vivo manipulation of mouse genomic sequences at the zygote stage. Proc. Natl. Acad. Sci. USA. 93, 5860-5865 http://dx.doi.org/10.1073/pnas.93.12.5860.
25. Li, Z., X. Cai, C.L. Cai, J. Wang, W. Zhang, B.E. Petersen, F.C. Yang, and M. Xu. 2011. Deletion of Tet2 in mice leads to dysregulated hematopoietic stem cells and subsequent development of myeloid malignancies. Blood. 118, 4509-4518 http://dx.doi.org/10.1182/blood-2010-12-325241.
26. Lin, Y.W., C. Slape, Z. Zhang, and P.D. Aplan. 2005. NUP98-HOXD13 transgenic mice develop a highly penetrant, severe myelodysplastic syndrome that progresses to acute leukemia. Blood. 106, 287-295 http://dx.doi.org/10.1182/blood-2004-12-4794.
27. Magini, P., M. Della Monica, M.L. Uzielli, P. Mongelli, G. Scarselli, E. Gambineri, G. Scarano, and M. Seri. 2012. Two novel patients with Bohring-Opitz syndrome caused by de novo ASXL1 mutations. Am. J. Med. Genet. A. 158A:917-921. http://dx.doi.org/10.1002/ajmg.a.35265
28. Metzeler, K.H., H. Becker, K. Maharry, M.D. Radmacher, J. Kohlschmidt, K. Mrózek, D. Nicolet, S.P. Whitman, Y.Z. Wu, S. Schwind, et al. 2011. ASXL1 mutations identify a high-risk subgroup of older patients with primary cytogenetically normal AML within the ELN Favorable genetic category. Blood. 118, 6920-6929 http://dx.doi.org/10.1182/blood-2011-08-368225.
29. Mizuki, M., J. Schwable, C. Steur, C. Choudhary, S. Agrawal, B. Sargin, B. Steffen, I. Matsumura, Y. Kanakura, F.D. Böhmer, et al. 2003. Suppression of myeloid transcription factors and induction of STAT response genes by AML-specific Flt3 mutations. Blood. 101, 3164-3173 http://dx.doi .org/10.1182/blood-2002-06-1677.
30. Moran-Crusio, K., L. Reavie, A. Shih, O. Abdel-Wahab, D. Ndiaye-Lobry, C. Lobry, M.E. Figueroa, A. Vasanthakumar, J. Patel, X. Zhao, et al. 2011. Tet2 loss leads to increased hematopoietic stem cell self-renewal and myeloid transformation. Cancer Cell. 20, 11-24 http://dx.doi.org/10 .1016/j.ccr.2011.06.001.
31. Mueller, B.U., T. Pabst, M. Osato, N. Asou, L.M. Johansen, M.D. Minden, G. Behre, W. Hiddemann, Y. Ito, and D.G. Tenen. 2002. Heterozygous PU.1 mutations are associated with acute myeloid leukemia. Blood. 100, 998-1007 http://dx.doi.org/10.1182/blood.V100.3.998.
32. Mullighan, C.G., A. Kennedy, X. Zhou, I. Radtke, L.A. Phillips, S.A. Shurtleff, and J.R. Downing. 2007. Pediatric acute myeloid leukemia with NPM1 mutations is characterized by a gene expression profile with dysregulated HOX gene expression distinct from MLL-rearranged leukemias. Leukemia. 21, 2000-2009 http://dx.doi.org/10.1038/sj.leu.2404808.
33. Nilsson, L., I. Astrand-Grundström, I. Arvidsson, B. Jacobsson, E. Hellström-Lindberg, R. Hast, and S.E. Jacobsen. 2000. Isolation and characterization of hematopoietic progenitor/stem cells in 5q-deleted myelodysplastic syndromes: evidence for involvement at the hematopoietic stem cell level. Blood. 96:2012-2021.
34. Nilsson, L., P. Edén, E. Olsson, R. Månsson, I. Astrand-Grundström, B. Strömbeck, K. Theilgaard-Mönch, K. Anderson, R. Hast, E. Hellström-Lindberg, et al. 2007. The molecular signature of MDS stem cells supports a stem-cell origin of 5q myelodysplastic syndromes. Blood. 110, 3005-3014 http://dx.doi.org/10.1182/blood-2007-03-079368.
35. Ohta, H., A. Sawada, J.Y. Kim, S. Tokimasa, S. Nishiguchi, R.K. Humphries, J. Hara, and Y. Takihara. 2002. Polycomb group gene rae28 is required for sustaining activity of hematopoietic stem cells. J. Exp. Med. 195, 759-770 http://dx.doi.org/10.1084/jem.20011911.
36. Pang, W.W., J.V. Pluvinage, E.A. Price, K. Sridhar, D.A. Arber, P.L. Greenberg, S.L. Schrier, C.Y. Park, and I.L. Weissman. 2013. Hematopoietic stem cell and progenitor cell mechanisms in myelodysplastic syndromes. Proc. Natl. Acad. Sci. USA. 110, 3011-3016 http://dx.doi.org/10.1073/pnas.1222861110.
37. Patel, J.P., M. Gönen, M.E. Figueroa, H. Fernandez, Z. Sun, J. Racevskis, P. Van Vlierberghe, I. Dolgalev, S. Thomas, O. Aminova, et al. 2012. Prognostic relevance of integrated genetic profiling in acute myeloid leukemia. N. Engl. J. Med. 366, 1079-1089 http://dx.doi.org/10.1056/NEJMoa1112304.
38. Quivoron, C., L. Couronné, V. Della Valle, C.K. Lopez, I. Plo, O. Wagner-Ballon, M. Do Cruzeiro, F. Delhommeau, B. Arnulf, M.H. Stern, et al. 2011. TET2 inactivation results in pleiotropic hematopoietic abnormalities in mouse and is a recurrent event during human lymphomagenesis. Cancer Cell. 20, 25-38 http://dx.doi.org/10.1016/j.ccr.2011.06.003.
39. Ramalho-Santos, M., S. Yoon, Y. Matsuzaki, R.C. Mulligan, and D.A. Melton. 2002. "Stemness": transcriptional profiling of embryonic and adult stem cells. Science. 298, 597-600 http://dx.doi.org/10.1126/science.1072530.
40. Raza-Egilmez, S.Z., S.N. Jani-Sait, M. Grossi, M.J. Higgins, T.B. Shows, and P.D. Aplan. 1998. NUP98-HOXD13 gene fusion in therapy-related acute myelogenous leukemia. Cancer Res. 58:4269-4273.
41. Rosenbauer, F., K. Wagner, J.L. Kutok, H. Iwasaki, M.M. Le Beau, Y. Okuno, K. Akashi, S. Fiering, and D.G. Tenen. 2004. Acute myeloid leukemia induced by graded reduction of a lineage-specific transcription factor, PU.1. Nat. Genet. 36, 624-630 http://dx.doi.org/10.1038/ng1361.
42. Ross, M.E., X. Zhou, G. Song, S.A. Shurtleff, K. Girtman, W.K. Williams, H.-C. Liu, R. Mahfouz, S.C. Raimondi, N. Lenny, et al. 2003. Classification of pediatric acute lymphoblastic leukemia by gene expression profiling. Blood. 102, 2951-2959 http://dx.doi.org/10.1182/blood-2003-01-0338.
43. Sanada, M., and S. Ogawa. 2012. Genome-wide analysis of myelodysplastic syndromes. Curr. Pharm. Des. 18, 3163-3169 http://dx.doi.org/10.2174/1381612811209023163.
44. Sawada, K., N. Sato, T. Tarumi, N. Sakai, K. Koizumi, S. Sakurama, M. Ieko, T. Yasukouchi, Y. Koyanagawa, M. Yamaguchi, et al. 1993. Proliferation and differentiation of myelodysplastic CD34+ cells in serum-free medium:response to individual colony-stimulating factors. Br. J. Haematol. 83, 349-358 http://dx.doi.org/10.1111/j.1365-2141.1993.tb04656.x.
45. Sawada, K., N. Sato, A. Notoya, T. Tarumi, S. Hirayama, H. Takano, K. Koizumi, T. Yasukouchi, M. Yamaguchi, and T. Koike. 1995. Proliferation and differentiation of myelodysplastic CD34+ cells: phenotypic subpopulations of marrow CD34+ cells. Blood. 85:194-202.
46. Shih, A.H., O. Abdel-Wahab, J.P. Patel, and R.L. Levine. 2012. The role of mutations in epigenetic regulators in myeloid malignancies. Nat. Rev. Cancer. 12, 599-612 http://dx.doi.org/10.1038/nrc3343.
47. Stadtfeld, M., and T. Graf. 2005. Assessing the role of hematopoietic plasticity for endothelial and hepatocyte development by non-invasive lineage tracing. Development. 132, 203-213 http://dx.doi.org/10.1242/dev.01558.
48. Steidl, U., F. Rosenbauer, R.G. Verhaak, X. Gu, A. Ebralidze, H.H. Otu, S. Klippel, C. Steidl, I. Bruns, D.B. Costa, et al. 2006. Essential role of Jun family transcription factors in PU.1 knockdown-induced leukemic stem cells. Nat. Genet. 38, 1269-1277 http://dx.doi.org/10.1038/ng1898.
49. Subramanian, A., P. Tamayo, V.K. Mootha, S. Mukherjee, B.L. Ebert, M.A. Gillette, A. Paulovich, S.L. Pomeroy, T.R. Golub, E.S. Lander, and J.P. Mesirov. 2005. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl. Acad. Sci. USA. 102, 15545-15550 http://dx.doi.org/10.1073/pnas.0506580102.
50. Tanaka, S., S. Miyagi, G. Sashida, T. Chiba, J. Yuan, M. Mochizuki-Kashio, Y. Suzuki, S. Sugano, C. Nakaseko, K. Yokote, et al. 2012. Ezh2 augments leukemogenicity by reinforcing differentiation blockage in acute myeloid leukemia. Blood. 120, 1107-1117 http://dx.doi .org/10.1182/blood-2011-11-394932.
51. Tehranchi, R., P.S. Woll, K. Anderson, N. Buza-Vidas, T. Mizukami, A.J. Mead, I. Astrand-Grundström, B. Strömbeck, A. Horvat, H. Ferry, et al. 2010. Persistent malignant stem cells in del(5q) myelodysplasia in remission. N. Engl. J. Med. 363, 1025-1037 http://dx.doi.org/10.1056/NEJMoa0912228.
52. Thol, F., I. Friesen, F. Damm, H. Yun, E.M. Weissinger, J. Krauter, K. Wagner, A. Chaturvedi, A. Sharma, M. Wichmann, et al. 2011. Prognostic significance of ASXL1 mutations in patients with myelodysplastic syndromes. J. Clin. Oncol. 29, 2499-2506 http://dx.doi.org/10.1200/JCO.2010.33.4938.
53. Vangala, R.K., M.S. Heiss-Neumann, J.S. Rangatia, S.M. Singh, C. Schoch, D.G. Tenen, W. Hiddemann, and G. Behre. 2003. The myeloid master regulator transcription factor PU.1 is inactivated by AML1-ETO in t(8;21) myeloid leukemia. Blood. 101, 270-277 http://dx.doi .org/10.1182/blood-2002-04-1288.
54. Vannucchi, A.M., T.L. Lasho, P. Guglielmelli, F. Biamonte, A. Pardanani, A. Pereira, C. Finke, J. Score, N. Gangat, C. Mannarelli, et al. 2013. Mutations and prognosis in primary myelofibrosis. Leukemia. 27, 1861-1869 http://dx.doi.org/10.1038/leu.2013.119.
55. Will, B., L. Zhou, T.O. Vogler, S. Ben-Neriah, C. Schinke, R. Tamari, Y. Yu, T.D. Bhagat, S. Bhattacharyya, L. Barreyro, et al. 2012. Stem and progenitor cells in myelodysplastic syndromes show aberrant stagespecific expansion and harbor genetic and epigenetic alterations. Blood. 120, 2076-2086 http://dx.doi.org/10.1182/blood-2011-12-399683.