Development Refractoriness of MLL-Rearranged Human B Cell Acute Leukemias to Reprogramming into Pluripotency.
Induced pluripotent stem cells (iPSCs) are a powerful tool for disease modeling. They are routinely generated from healthy donors and patients from multiple cell types at different developmental stages. However, reprogramming leukemias is an extremely inefficient process. Few studies generated iPSCs from primary chronic myeloid leukemias, but iPSC generation from acute myeloid or lymphoid leukemias (ALL) has not been achieved. We attempted to generate iPSCs from different subtypes of B-ALL to address the developmental impact of leukemic fusion genes. OKSM(L)-expressing mono/polycistronic-, retroviral/lentiviral/episomal-, and Sendai virus vector-based reprogramming strategies failed to render iPSCs in vitro and in vivo. Addition of transcriptomic-epigenetic reprogramming "boosters" also failed to generate iPSCs from B cell blasts and B-ALL lines, and when iPSCs emerged they lacked leukemic fusion genes, demonstrating non-leukemic myeloid origin. Conversely, MLL-AF4-overexpressing hematopoietic stem cells/B progenitors were successfully reprogrammed, indicating that B cell origin and leukemic fusion gene were not reprogramming barriers. Global transcriptome/DNA methylome profiling suggested a developmental/differentiation refractoriness of MLL-rearranged B-ALL to reprogramming into pluripotency.
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Department of Biomedicine, School of Medicine, University of Barcelona, 08036 Barcelona, Spain.
Alvaro
A
Muñoz-López
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Department of Biomedicine, School of Medicine, University of Barcelona, 08036 Barcelona, Spain.
Damià
D
Romero-Moya
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Department of Biomedicine, School of Medicine, University of Barcelona, 08036 Barcelona, Spain.
Cristina
C
Prieto
Genomic Oncology Department, Centre for Genomics and Oncology GENyO, 18016 Granada, Spain.
Verónica
V
Ramos-Mejía
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Department of Biomedicine, School of Medicine, University of Barcelona, 08036 Barcelona, Spain; IBBTEC, CSIC-University of Cantabria, 39011 Santander, Spain.
Antonio
A
Agraz-Doblas
IBBTEC, CSIC-University of Cantabria, 39011 Santander, Spain.
Ignacio
I
Varela
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain.
Marcus
M
Buschbeck
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain.
Anna
A
Palau
Cell Therapy Department, Centro de Investigación Médica Aplicada (CIMA), 31008 Pamplona, Spain.
Xonia
X
Carvajal-Vergara
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain.
Alessandra
A
Giorgetti
Centre for Evolution and Cancer, Institute of Cancer Research, London SW7 3RP, UK.
Anthony
A
Ford
Institute of Genetic Medicine, Newcastle University, Newcastle NE1 7RU, UK.
Majlinda
M
Lako
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Hematology Department, Hospital Germans Trias i Pujol, Institut Català d'Oncología, 08916 Badalona, Spain.
Isabel
I
Granada
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Hematology Department, Hospital Germans Trias i Pujol, Institut Català d'Oncología, 08916 Badalona, Spain.
Neus
N
Ruiz-Xivillé
Cytogenetics Group, Centro Nacional de Investigaciones Oncológicas (CNIO), 28029 Madrid, Spain.
Sandra
S
Rodríguez-Perales
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Cytogenetics Group, Centro Nacional de Investigaciones Oncológicas (CNIO), 28029 Madrid, Spain.
Raul
R
Torres-Ruíz
Department of Pediatric Oncology/Hematology, Erasmus Medical Center, Erasmus University, 3015 CN Rotterdam, the Netherlands.
Ronald W
RW
Stam
Department of Pediatric Oncohematology, Clinical University Hospital Virgen de la Arrixaca, 30120 Murcia, Spain.
Jose Luis
JL
Fuster
Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA-HUCA), Universidad de Oviedo, 33003 Oviedo, Spain.
Mario F
MF
Fraga
Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraka 305-0046, Japan.
Mahito
M
Nakanishi
University di Milano-Bicocca, Ospedale San Gerardo/Fondazione MBBM, 20052 Monza MB, Italy.
Gianni
G
Cazzaniga
University di Milano-Bicocca, Ospedale San Gerardo/Fondazione MBBM, 20052 Monza MB, Italy.
Michela
M
Bardini
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA-HUCA), Universidad de Oviedo, 33003 Oviedo, Spain.
Isabel
I
Cobo
Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA-HUCA), Universidad de Oviedo, 33003 Oviedo, Spain.
Gustavo F
GF
Bayon
Cancer Epigenetics Laboratory, Instituto Universitario de Oncología del Principado de Asturias (IUOPA-HUCA), Universidad de Oviedo, 33003 Oviedo, Spain.
Agustin F
AF
Fernandez
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Department of Biomedicine, School of Medicine, University of Barcelona, 08036 Barcelona, Spain. Electronic address: cbueno@carrerasresearch.org.
Clara
C
Bueno
Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Casanova 143, 08036 Barcelona, Spain; Department of Biomedicine, School of Medicine, University of Barcelona, 08036 Barcelona, Spain; Instituciò Catalana de Recerca i Estudis Avançats (ICREA), 08036 Barcelona, Spain. Electronic address: pmenendez@carrerasresearch.org.
Pablo
P
Menendez
D000818:Animals; D015415:Biomarkers; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D054338:Cell Transdifferentiation / Q000235:genetics*; D065150:Cellular Reprogramming; D016000:Cluster Analysis; D019175:DNA Methylation; D015870:Gene Expression; D020869:Gene Expression Profiling; D015321:Gene Rearrangement; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D064593:Heterografts; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D023461:Myeloid Progenitor Cells / Q000378:metabolism; D051788:Myeloid-Lymphoid Leukemia Protein / Q000235:genetics*; D015514:Oncogene Proteins, Fusion / Q000235:genetics; D010641:Phenotype; D015452:Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics*; D054448:Precursor Cells, B-Lymphoid / Q000378:metabolism; D059467:Transcriptome; D014178:Translocation, Genetic
D015415:Biomarkers; C113855:MLL-AF4 fusion protein, human; D015514:Oncogene Proteins, Fusion; D051788:Myeloid-Lymphoid Leukemia Protein
B-ALL; DNA methylome; MLL-AF4; Sendai virus; cancer reprogramming; iPSC; transcriptome
EMBO J. 2011 May 06;30(12):2373-87
21552206
Haematologica. 2014 Jul;99(7):1168-75
24727813
Leukemia. 2014 Mar;28(3):666-74
24240202
Nature. 2014 Mar 6;507(7490):99-103
24413397
Leukemia. 2008 Aug;22(8):1475-8
18698324
Cell Cycle. 2010 Oct 1;9(19):3887-91
20948296
Mol Ther. 2013 Jun;21(6):1242-50
23568261
Stem Cells Int. 2015;2015:794632
25922608
Mol Ther. 2012 Jul;20(7):1443-53
22491213
PLoS One. 2015 Mar 24;10 (3):e0120841
25803811
Cell Stem Cell. 2016 Feb 4;18(2):262-75
26686465
Cancer Res. 2007 Feb 1;67(3):876-80
17283117
Stem Cells. 2016 Mar;34(3):581-7
26850912
Stem Cells Dev. 2014 Jun 15;23(12):1285-300
24524728
Immunol Rev. 2000 Aug;176:30-46
11043766
Cell. 2009 Oct 30;139(3):573-86
19879843
PLoS One. 2013 Dec 12;8(12 ):e82838
24349377
Nat Commun. 2013;4:1565
23463008
Proc Natl Acad Sci U S A. 2005 May 24;102(21):7635-40
15897469
PLoS One. 2012;7(4):e35824
22545141
Stem Cells Dev. 2013 Aug 15;22(16):2268-77
23534850
Blood. 2009 Dec 24;114(27):5473-80
19797525
Nature. 2014 Feb 13;506(7487):235-9
24336202
Cancer Lett. 2015 Dec 1;369(1):1-8
26276716
Leukemia. 2016 Jul;30(7):1603-6
26710885
Nat Commun. 2014 Jun 03;5:3964
24888982
J Exp Med. 2009 Dec 21;206(13):3131-41
19995953
Mol Cell. 2010 May 28;38(4):576-89
20513432
Nat Cell Biol. 2011 May;13(5):497-505
21540845
Proc Natl Acad Sci U S A. 2015 Mar 31;112(13):4074-9
25775523
Stem Cells Transl Med. 2014 Dec;3(12):1429-34
25298370
Cancer Res. 2016 Apr 15;76(8):2478-89
26837759
Blood. 2012 Jun 28;119(26):6234-42
22592606
Mol Cancer. 2008 Oct 03;7:76
18834512
Genomics. 2011 Oct;98(4):288-95
21839163
Nat Cell Biol. 2015 Aug;17(8):1004-13
26214134
Stem Cells Dev. 2012 May 1;21(7):1145-55
21671728
Leukemia. 2016 Feb;30(2):508-13
26084867
World J Stem Cells. 2015 Mar 26;7(2):461-9
25815129
Nature. 2012 Mar 04;483(7391):598-602
22388813
Cell Stem Cell. 2010 Jan 8;6(1):71-9
20036631
Blood. 2010 May 20;115(20):4039-42
20233975
Trends Mol Med. 2012 May;18(5):245-7
22521522
Blood. 2013 Jun 13;121(24):4925-9
23620576
Cell Res. 2009 Jun;19(6):698-709
19308090
PLoS One. 2013 Aug 23;8(8):e71596
24058405
Carcinogenesis. 2009 Sep;30(9):1628-37
19587093
Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5116-21
11309499
Leukemia. 2016 Mar;30(3):674-82
26500142
Cell Mol Life Sci. 2013 Jul;70(14):2443-61
23052214
EMBO J. 2015 May 12;34(10):1296-308
25820261
Cytotherapy. 2006;8(6):530-41
17148029
Nucleic Acids Res. 2003 Feb 15;31(4):e15
12582260
Stem Cells Dev. 2012 Jun 10;21(9):1549-58
21936705
Blood. 2014 Nov 13;124(20):3065-75
25185710