Identification of CD34+ and CD34- leukemia-initiating cells in MLL-rearranged human acute lymphoblastic leukemia.

Translocation of the mixed-lineage leukemia (MLL) gene with AF4, AF9, or ENL results in acute leukemia with both lymphoid and myeloid involvement. We characterized leukemia-initiating cells (LICs) in primary infant MLL-rearranged leukemia using a xenotransplantation model. In MLL-AF4 patients, CD34(+)CD38(+)CD19(+) and CD34(-)CD19(+) cells initiated leukemia, and in MLL-AF9 patients, CD34(-)CD19(+) cells were LICs. In MLL-ENL patients, either CD34(+) or CD34(-) cells were LICs, depending on the pattern of CD34 expression. In contrast, in patients with these MLL translocations, CD34(+)CD38(-)CD19(-)CD33(-) cells were enriched for normal hematopoietic stem cells (HSCs) with in vivo long-term multilineage hematopoietic repopulation capacity. Although LICs developed leukemic cells with clonal immunoglobulin heavy-chain (IGH) rearrangement in vivo, CD34(+)CD38(-)CD19(-)CD33(-) cells repopulated recipient bone marrow and spleen with B cells, showing broad polyclonal IGH rearrangement and recipient thymus with CD4(+) single positive (SP), CD8(+) SP, and CD4(+)CD8(+) double-positive (DP) T cells. Global gene expression profiling revealed that CD9, CD32, and CD24 were over-represented in MLL-AF4, MLL-AF9, and MLL-ENL LICs compared with normal HSCs. In patient samples, these molecules were expressed in CD34(+)CD38(+) and CD34(-) LICs but not in CD34(+)CD38(-)CD19(-)CD33(-) HSCs. Identification of LICs and LIC-specific molecules in primary human MLL-rearranged acute lymphoblastic leukemia may lead to improved therapeutic strategies for MLL-rearranged leukemia.

[1]  Yi Zheng,et al.  Microenvironment determines lineage fate in a human model of MLL-AF9 leukemia. , 2008, Cancer cell.

[2]  R. Balderas,et al.  Repression of the genome organizer SATB1 in regulatory T cells is required for suppressive function and inhibition of effector differentiation , 2011, Nature Immunology.

[3]  S. Iwata,et al.  CD9 correlates with cancer stem cell potentials in human B-acute lymphoblastic leukemia cells. , 2009, Biochemical and biophysical research communications.

[4]  A. Perkins,et al.  Homeobox gene expression plus autocrine growth factor production elicits myeloid leukemia. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[5]  K. Anderson,et al.  Distinct patterns of hematopoietic stem cell involvement in acute lymphoblastic leukemia , 2005, Nature Medicine.

[6]  J. Rowley,et al.  International workshop on the relationship of prior therapy to balanced chromosome aberrations in therapy‐related myelodysplastic syndromes and acute leukemia: Overview report , 2002, Genes, chromosomes & cancer.

[7]  Michael R. Green,et al.  Initiation of zebrafish haematopoiesis by the TATA-box-binding protein-related factor Trf3 , 2007, Nature.

[8]  Brown Mh,et al.  The SIRP family of receptors and immune regulation. , 2006 .

[9]  A. Ullrich,et al.  Human signal-regulatory protein is expressed on normal, but not on subsets of leukemic myeloid cells and mediates cellular adhesion involving its counterreceptor CD47. , 1999, Blood.

[10]  T. Kohwi-Shigematsu,et al.  SATB1 packages densely looped, transcriptionally active chromatin for coordinated expression of cytokine genes , 2006, Nature Genetics.

[11]  T. K. van den Berg,et al.  Signal-regulatory protein is selectively expressed by myeloid and neuronal cells. , 1998, Journal of immunology.

[12]  Yi Zhang,et al.  DOT1L, the H3K79 methyltransferase, is required for MLL-AF9-mediated leukemogenesis. , 2011, Blood.

[13]  M. Ono,et al.  Augmented humoral and anaphylactic responses in FcγRII-deficient mice , 1996, Nature.

[14]  Akihiko Yokoyama,et al.  Menin critically links MLL proteins with LEDGF on cancer-associated target genes. , 2008, Cancer cell.

[15]  Rainer Breitling,et al.  RankProd: a bioconductor package for detecting differentially expressed genes in meta-analysis , 2006, Bioinform..

[16]  R. Pieters,et al.  A treatment protocol for infants younger than 1 year with acute lymphoblastic leukaemia (Interfant-99): an observational study and a multicentre randomised trial , 2007, The Lancet.

[17]  Yi Zhang,et al.  hDOT1L Links Histone Methylation to Leukemogenesis , 2005, Cell.

[18]  D. Maloney.  Anti-CD20 antibody therapy for B-cell lymphomas. , 2012, The New England journal of medicine.

[19]  M. Kotb,et al.  Human Lymphoid and Myeloid Cell Development in NOD/LtSz-scid IL2Rγnull Mice Engrafted with Mobilized Human Hemopoietic Stem Cells 12 , 2004, The Journal of Immunology.

[20]  Verena I Gaidzik,et al.  TET2 mutations in acute myeloid leukemia (AML): results from a comprehensive genetic and clinical analysis of the AML study group. , 2012, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[21]  S. Korsmeyer,et al.  Definitive hematopoiesis requires the mixed-lineage leukemia gene. , 2004, Developmental cell.

[22]  M. Eguchi,et al.  Outcome of risk-based therapy for infant acute lymphoblastic leukemia with or without an MLL gene rearrangement, with emphasis on late effects: a final report of two consecutive studies, MLL96 and MLL98, of the Japan Infant Leukemia Study Group , 2007, Leukemia.

[23]  Chuan He,et al.  Tet Proteins Can Convert 5-Methylcytosine to 5-Formylcytosine and 5-Carboxylcytosine , 2011, Science.

[24]  M Hummel,et al.  Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: Report of the BIOMED-2 Concerted Action BMH4-CT98-3936 , 2003, Leukemia.

[25]  E. Bertolino,et al.  Transcription factor EBF restricts alternative lineage options and promotes B cell fate commitment independently of Pax5 , 2008, Nature Immunology.

[26]  A. Gewirtz,et al.  c-Myb binds MLL through menin in human leukemia cells and is an important driver of MLL-associated leukemogenesis. , 2010, The Journal of clinical investigation.

[27]  Rob Pieters,et al.  In childhood acute lymphoblastic leukemia, blasts at different stages of immunophenotypic maturation have stem cell properties. , 2008, Cancer cell.

[28]  R. Wenger,et al.  Altered erythrocytes and a leaky block in B-cell development in CD24/HSA-deficient mice. , 1997, Blood.

[29]  David R. Liu,et al.  Conversion of 5-Methylcytosine to 5- Hydroxymethylcytosine in Mammalian DNA by the MLL Partner TET1 , 2009 .

[30]  Peng Fei Wang,et al.  Molecular Regulation of H3K4 Trimethylation by Wdr82, a Component of Human Set1/COMPASS , 2008, Molecular and Cellular Biology.

[31]  Xiaobo Xia,et al.  H3K79 methylation profiles define murine and human MLL-AF4 leukemias. , 2008, Cancer cell.

[32]  Satoshi Tanaka,et al.  Chemotherapy-resistant human AML stem cells home to and engraft within the bone-marrow endosteal region , 2007, Nature Biotechnology.

[33]  C. So,et al.  β-Catenin mediates the establishment and drug resistance of MLL leukemic stem cells. , 2010, Cancer cell.

[34]  J. Dick,et al.  Disruption of SIRPα signaling in macrophages eliminates human acute myeloid leukemia stem cells in xenografts , 2012, The Journal of experimental medicine.

[35]  C. Pilarsky,et al.  CD24 is expressed in ovarian cancer and is a new independent prognostic marker of patient survival. , 2002, The American journal of pathology.

[36]  R. Stam,et al.  Gene expression profiling-based dissection of MLL translocated and MLL germline acute lymphoblastic leukemia in infants. , 2010, Blood.

[37]  N. Socci,et al.  Prognostic relevance of integrated genetic profiling in acute myeloid leukemia. , 2012, The New England journal of medicine.

[38]  A. Barclay,et al.  The SIRP family of receptors and immune regulation , 2006, Nature Reviews Immunology.

[39]  Howard Y. Chang,et al.  Identification, molecular characterization, clinical prognosis, and therapeutic targeting of human bladder tumor-initiating cells , 2009, Proceedings of the National Academy of Sciences.

[40]  Ash A. Alizadeh,et al.  Therapeutic antibody targeting of CD47 eliminates human acute lymphoblastic leukemia. , 2011, Cancer research.

[41]  K. Akashi,et al.  Development of functional human blood and immune systems in NOD/SCID/IL2 receptor γ chainnull mice , 2005 .

[42]  J. Zuber,et al.  A reconfigured pattern of MLL occupancy within mitotic chromatin promotes rapid transcriptional reactivation following mitotic exit. , 2009, Molecular cell.

[43]  Rafael A. Irizarry,et al.  A Model-Based Background Adjustment for Oligonucleotide Expression Arrays , 2004 .

[44]  J. Hsieh,et al.  MLL fusions: Pathways to leukemia , 2009, Cancer biology & therapy.

[45]  R. Grosschedl,et al.  Failure of B-cell differentiation in mice lacking the transcription factor EBF , 1995, Nature.

[46]  D. Campana,et al.  Age-related differences in leukemia biology and prognosis: the paradigm of MLL-AF4-positive acute lymphoblastic leukemia , 2007, Leukemia.

[47]  Bernd Hauck,et al.  Chimeric antigen receptor-modified T cells for acute lymphoid leukemia. , 2013, The New England journal of medicine.

[48]  Giacomo Cavalli,et al.  Trithorax group proteins: switching genes on and keeping them active , 2011, Nature Reviews Molecular Cell Biology.

[49]  O. Ohara,et al.  Identification of Therapeutic Targets for Quiescent, Chemotherapy-Resistant Human Leukemia Stem Cells , 2010, Science Translational Medicine.

[50]  L. Zon,et al.  An Mll-Dependent Hox Program Drives Hematopoietic Progenitor Expansion , 2004, Current Biology.

[51]  Jens-Peter Volkmer,et al.  The CD47-signal regulatory protein alpha (SIRPa) interaction is a therapeutic target for human solid tumors , 2012, Proceedings of the National Academy of Sciences.

[52]  M. Kasuga,et al.  A novel membrane glycoprotein, SHPS-1, that binds the SH2-domain-containing protein tyrosine phosphatase SHP-2 in response to mitogens and cell adhesion , 1996, Molecular and cellular biology.

[53]  Yang Wang,et al.  Tet-Mediated Formation of 5-Carboxylcytosine and Its Excision by TDG in Mammalian DNA , 2011, Science.

[54]  Takuro Nakamura,et al.  Cooperative activation of Hoxa and Pbx1-related genes in murine myeloid leukaemias , 1996, Nature Genetics.

[55]  K. Maruyama,et al.  Deletion of Tetraspanin CD9 Diminishes Lymphangiogenesis in Vivo and in Vitro* , 2012, The Journal of Biological Chemistry.

[56]  A. Kohlmann,et al.  Landscape of TET2 mutations in acute myeloid leukemia , 2012, Leukemia.

[57]  A. Feinberg,et al.  Fusion of the nucleoporin gene NUP98 to HOXA9 by the chromosome translocation t(7;11)(p15;p15) in human myeloid leukaemia , 1996, Nature Genetics.

[58]  T. Schlange,et al.  CD24 controls Src/STAT3 activity in human tumors , 2012, Cellular and Molecular Life Sciences.

[59]  X. Hua,et al.  In search of tumor suppressing functions of menin , 2007, Molecular and Cellular Endocrinology.

[60]  J. Chen,et al.  Association between the Polymorphism rs3217927 of CCND2 and the Risk of Childhood Acute Lymphoblastic Leukemia in a Chinese Population , 2014, PloS one.

[61]  K. Akashi,et al.  Development of functional human blood and immune systems in NOD/SCID/IL2 receptor {gamma} chain(null) mice. , 2005, Blood.

[62]  J. Fisher,et al.  Unique and independent roles for MLL in adult hematopoietic stem cells and progenitors. , 2007, Cell stem cell.

[63]  Hui Zhang,et al.  MLL-AF9-induced leukemogenesis requires coexpression of the wild-type Mll allele. , 2010, Cancer cell.

[64]  Todd D. Westergard,et al.  Phosphorylation of MLL by ATR is Required for Execution of Mammalian S Phase Checkpoint , 2010, Nature.

[65]  A. Ullrich,et al.  A family of proteins that inhibit signalling through tyrosine kinase receptors , 1997, Nature.

[66]  N. Heerema,et al.  Analysis of prognostic factors of acute lymphoblastic leukemia in infants: report on CCG 1953 from the Children's Oncology Group. , 2006, Blood.

[67]  Masato Tanaka,et al.  Construction of an open-access database that integrates cross-reference information from the transcriptome and proteome of immune cells , 2007, Bioinform..

[68]  Gordon K Smyth,et al.  Statistical Applications in Genetics and Molecular Biology Linear Models and Empirical Bayes Methods for Assessing Differential Expression in Microarray Experiments , 2011 .

[69]  Andrew H. Beck,et al.  Antibody therapy targeting the CD47 protein is effective in a model of aggressive metastatic leiomyosarcoma , 2012, Proceedings of the National Academy of Sciences.

[70]  Nadia Passon,et al.  Histone post-translational modifications associated to BAALC expression in leukemic cells. , 2012, Biochemical and biophysical research communications.

[71]  N. Goulden,et al.  Characterization of acute lymphoblastic leukemia progenitor cells. , 2004, Blood.

[72]  T. Sasaki,et al.  Identification of Zfp521/ZNF521 as a cooperative gene for E2A-HLF to develop acute B-lineage leukemia , 2010, Oncogene.

[73]  Dafydd G. Thomas,et al.  Requirement for Dot1l in murine postnatal hematopoiesis and leukemogenesis by MLL translocation. , 2011, Blood.

[74]  C. Pilarsky,et al.  CD24 expression is a new prognostic marker in breast cancer. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[75]  M. Ono,et al.  Augmented humoral and anaphylactic responses in Fc gamma RII-deficient mice. , 1996, Nature.

[76]  Verena I Gaidzik,et al.  RUNX1 mutations in acute myeloid leukemia: results from a comprehensive genetic and clinical analysis from the AML study group. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[77]  M. Cleary,et al.  Immortalization and leukemic transformation of a myelomonocytic precursor by retrovirally transduced HRX–ENL , 1997, The EMBO journal.

[78]  Nikolaus Rajewsky,et al.  Modeling the Initiation and Progression of Human Acute Leukemia in Mice , 2022 .