A distinct subpopulation of leukemia initiating cells in acute precursor B lymphoblastic leukemia: quiescent phenotype and unique transcriptomic profile

In leukemia, a distinct subpopulation of cancer-initiating cells called leukemia stem cells (LSCs) is believed to drive population expansion and tumor growth. Failing to eliminate LSCs may result in disease relapse regardless of the amount of non-LSCs destroyed. The first step in targeting and eliminating LSCs is to identify and characterize them. Acute precursor B lymphoblastic leukemia (B-ALL) cells derived from patients were incubated with fluorescent glucose analog 2-(N-(7-Nitrobenz-2-oxa-1, 3-diazol-4-yl) Amino)-2-Deoxyglucose (NBDG) and sorted based on NBDG uptake. Cell subpopulations defined by glucose uptake were then serially transplanted into mice and evaluated for leukemia initiating capacity. Gene expression profiles of these cells were characterized using RNA-Sequencing (RNA-Seq). A distinct population of NBDG-low cells was identified in patient B-ALL samples. These cells are a small population (1.92% of the entire leukemia population), have lower HLA expression, and are smaller in size (4.0 to 7.0 μm) than the rest of the leukemia population. All mice transplanted with NBDG-low cells developed leukemia between 5 and 14 weeks, while those transplanted with NBDG-high cells did not develop leukemia (p ≤ 0.0001-0.002). Serial transplantation of the NBDG-low mouse model resulted in successful leukemia development. NBDG-medium (NBDG-med) populations also developed leukemia. Interestingly, comprehensive molecular characterization of NBDG-low and NBDG-med cells from patient-derived xenograft (PDX) models using RNA-Seq revealed a distinct profile of 2,162 differentially-expressed transcripts (DETs) (p<0.05) with 70.6% down-regulated in NBDG-low cells. Hierarchical clustering of DETs showed distinct segregation of NBDG-low from NBDG-med and NBDG-high groups with marked transcription expression alterations in the NBDG-low group consistent with cancer survival. In conclusion, A unique subpopulation of cells with low glucose uptake (NBDG-low) in B-ALL was discovered. These cells, despite their quiescence characteristics, once transplanted in mice, showed potent leukemia initiating capacity. Although NBDG-med cells also initiated leukemia, gene expression profiling revealed a distinct signature that clearly distinguishes NBDG-low cells from NBDG-med and the rest of the leukemia populations. These results suggest that NBDG-low cells may represent quiescent LSCs. These cells can be activated in the appropriate environment in vivo, showing leukemia initiating capacity. Our study provides insight into the biologic mechanisms of B-ALL initiation and survival.

[1]  Huihui Liu,et al.  Lipid metabolism of cancer stem cells , 2022, Oncology letters.

[2]  Gary D Bader,et al.  The reactome pathway knowledgebase 2022 , 2021, Nucleic Acids Res..

[3]  B. Beutler,et al.  Emerging roles of spliceosome in cancer and immunity , 2021, Protein & Cell.

[4]  S. Ogawa,et al.  Chromatin-Spliceosome Mutations in Acute Myeloid Leukemia , 2021, Cancers.

[5]  Fan Yang,et al.  The regulation of protein translation and its implications for cancer , 2021, Signal Transduction and Targeted Therapy.

[6]  Anushya Muruganujan,et al.  The Gene Ontology resource: enriching a GOld mine , 2020, Nucleic Acids Res..

[7]  Y. Dou,et al.  ER associated degradation preserves hematopoietic stem cell quiescence and self-renewal by restricting mTOR activity. , 2020, Blood.

[8]  E. Vanzyl,et al.  The spliceosome inhibitors isoginkgetin and pladienolide B induce ATF3-dependent cell death , 2019, bioRxiv.

[9]  Justin Taylor,et al.  Mutations in spliceosome genes and therapeutic opportunities in myeloid malignancies , 2019, Genes, chromosomes & cancer.

[10]  T. Rando,et al.  Stem Cell Quiescence: Dynamism, Restraint, and Cellular Idling. , 2019, Cell stem cell.

[11]  Y. Ho,et al.  HDAC inhibitor suppresses proliferation and tumorigenicity of drug‐resistant chronic myeloid leukemia stem cells through regulation of hsa‐miR‐196a targeting BCR/ABL1 , 2018, Experimental cell research.

[12]  S. Wingett,et al.  FastQ Screen: A tool for multi-genome mapping and quality control. , 2018, F1000Research.

[13]  R. Mancini,et al.  Metabolic features of cancer stem cells: the emerging role of lipid metabolism , 2018, Oncogene.

[14]  Matthew R. Gazzara,et al.  Aberrant splicing in B-cell acute lymphoblastic leukemia , 2017, bioRxiv.

[15]  C. Sette,et al.  Alternative splicing and cell survival: from tissue homeostasis to disease , 2016, Cell Death and Differentiation.

[16]  Yangqiu Li,et al.  Heterogeneity of CD34 and CD38 expression in acute B lymphoblastic leukemia cells is reversible and not hierarchically organized , 2016, Journal of Hematology & Oncology.

[17]  F. Sotgia,et al.  Cancer stem cell metabolism , 2016, Breast Cancer Research.

[18]  S. Minucci,et al.  Inhibition of histone deacetylases in cancer therapy: lessons from leukaemia , 2016, British Journal of Cancer.

[19]  A. Syme,et al.  Phosphorylation of eIF2α Is a Translational Control Mechanism Regulating Muscle Stem Cell Quiescence and Self-Renewal. , 2016, Cell stem cell.

[20]  Minoru Kanehisa,et al.  KEGG as a reference resource for gene and protein annotation , 2015, Nucleic Acids Res..

[21]  David Allman,et al.  Convergence of Acquired Mutations and Alternative Splicing of CD19 Enables Resistance to CART-19 Immunotherapy. , 2015, Cancer discovery.

[22]  Enric Llorens-Bobadilla,et al.  Single-Cell Transcriptomics Reveals a Population of Dormant Neural Stem Cells that Become Activated upon Brain Injury. , 2015, Cell stem cell.

[23]  M. Rieger,et al.  Stem Cell Hierarchy and Clonal Evolution in Acute Lymphoblastic Leukemia , 2015, Stem cells international.

[24]  J. Li,et al.  A New Strategy to Target Acute Myeloid Leukemia Stem and Progenitor Cells Using Chidamide, a Histone Deacetylase Inhibitor. , 2015, Current cancer drug targets.

[25]  C. Schürch,et al.  Regulation of hematopoietic and leukemic stem cells by the immune system , 2014, Cell Death and Differentiation.

[26]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[27]  S. Morrison,et al.  Cellular Differences in Protein Synthesis Regulate Tissue Homeostasis , 2014, Cell.

[28]  W. Du,et al.  Binding to WGR domain by salidroside activates PARP1 and protects hematopoietic stem cells from oxidative stress. , 2014, Antioxidants & redox signaling.

[29]  Jeffrey A. Magee,et al.  Haematopoietic stem cells require a highly regulated protein synthesis rate , 2014, Nature.

[30]  M. Olivotto,et al.  Hypoxia-resistant profile implies vulnerability of cancer stem cells to physiological agents, which suggests new therapeutic targets , 2013, Cell cycle.

[31]  M. Teitell,et al.  Techniques to monitor glycolysis. , 2014, Methods in enzymology.

[32]  J. Haybaeck,et al.  Eukaryotic translation initiation factors in cancer development and progression. , 2013, Cancer letters.

[33]  John M. Ashton,et al.  BCL-2 inhibition targets oxidative phosphorylation and selectively eradicates quiescent human leukemia stem cells. , 2013, Cell stem cell.

[34]  Thomas R. Gingeras,et al.  STAR: ultrafast universal RNA-seq aligner , 2013, Bioinform..

[35]  W. Du,et al.  Salidroside stimulates DNA repair enzyme Parp-1 activity in mouse HSC maintenance. , 2012, Blood.

[36]  H. Coller,et al.  Staying alive , 2012, Cell cycle.

[37]  H. Boswell,et al.  Novel combination treatments targeting chronic myeloid leukemia stem cells. , 2012, Clinical lymphoma, myeloma & leukemia.

[38]  Stephen L. Abrams,et al.  Targeting the translational apparatus to improve leukemia therapy: roles of the PI3K/PTEN/Akt/mTOR pathway , 2011, Leukemia.

[39]  D. Charron,et al.  Age‐related changes in human hematopoietic stem/progenitor cells , 2011, Aging cell.

[40]  M. Assanah,et al.  HnRNP proteins controlled by c-Myc deregulate pyruvate kinase mRNA splicing in cancer , 2010, Nature.

[41]  I. Sánchez-García,et al.  B‐cell acute lymphoblastic leukaemia: towards understanding its cellular origin , 2009, BioEssays : news and reviews in molecular, cellular and developmental biology.

[42]  Timothy J. Muldoon,et al.  Molecular imaging of glucose uptake in oral neoplasia following topical application of fluorescently labeled deoxy‐glucose , 2009, International journal of cancer.

[43]  Jing Chen,et al.  ToppGene Suite for gene list enrichment analysis and candidate gene prioritization , 2009, Nucleic Acids Res..

[44]  W. Evans,et al.  Inhibition of glycolysis modulates prednisolone resistance in acute lymphoblastic leukemia cells. , 2009, Blood.

[45]  Y. Liu,et al.  p53 regulates hematopoietic stem cell quiescence. , 2009, Cell stem cell.

[46]  Scott A Armstrong,et al.  Leukemia stem cells and human acute lymphoblastic leukemia. , 2009, Seminars in hematology.

[47]  M. Baker Cancer stem cells, becoming common , 2008 .

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

[49]  M. Loh,et al.  Risk- and response-based classification of childhood B-precursor acute lymphoblastic leukemia: a combined analysis of prognostic markers from the Pediatric Oncology Group (POG) and Children's Cancer Group (CCG). , 2007, Blood.

[50]  Chi V Dang,et al.  Cancer's molecular sweet tooth and the Warburg effect. , 2006, Cancer research.

[51]  Martin J Firth,et al.  Altered glucose metabolism in childhood pre-B acute lymphoblastic leukaemia , 2006, Leukemia.

[52]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[53]  Dagmar Ringe,et al.  “Sleeping Beauty”: Quiescence in Saccharomyces cerevisiae , 2004, Microbiology and Molecular Biology Reviews.

[54]  M. Trigg Hematopoietic stem cells. , 2004, Pediatrics.

[55]  Ioannis S Vizirianakis,et al.  Mechanisms involved in the induced differentiation of leukemia cells. , 2003, Pharmacology & therapeutics.

[56]  M. D. Boer,et al.  Patient stratification based on prednisolone-vincristine-asparaginase resistance profiles in children with acute lymphoblastic leukemia. , 2003, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[57]  T. Flores,et al.  A primitive hematopoietic cell is the target for the leukemic transformation in human philadelphia-positive acute lymphoblastic leukemia. , 2000, Blood.

[58]  K. Moore,et al.  Single adult human CD34(+)/Lin-/CD38(-) progenitors give rise to natural killer cells, B-lineage cells, dendritic cells, and myeloid cells. , 1999, Blood.

[59]  Ph,et al.  Primordial role of CD34+38− cells in early and late trilineage haemopoietic engraftment after autologous blood cell transplantation , 1998, British journal of haematology.

[60]  R. Pieters,et al.  Prednisolone resistance in childhood acute lymphoblastic leukemia: vitro-vivo correlations and cross-resistance to other drugs. , 1998, Blood.

[61]  L. Ailles,et al.  Detection and characterization of primitive malignant and normal progenitors in patients with acute myelogenous leukemia using long-term coculture with supportive feeder layers and cytokines. , 1997, Blood.

[62]  J. Dick,et al.  Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell , 1997, Nature Medicine.

[63]  T. Hongo,et al.  In vitro drug sensitivity testing can predict induction failure and early relapse of childhood acute lymphoblastic leukemia. , 1997, Blood.

[64]  M. Caligiuri,et al.  A cell initiating human acute myeloid leukaemia after transplantation into SCID mice , 1994, Nature.

[65]  C. Jordan,et al.  Primitive hemopoietic stem cells: direct assay of most productive populations by competitive repopulation with simple binomial, correlation and covariance calculations. , 1993, Experimental hematology.

[66]  A. Eaves,et al.  Rapid decline of chronic myeloid leukemic cells in long-term culture due to a defect at the leukemic stem cell level. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[67]  C. Eaves,et al.  Quantitative assay for totipotent reconstituting hematopoietic stem cells by a competitive repopulation strategy. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[68]  D. Harrison Competitive repopulation: a new assay for long-term stem cell functional capacity. , 1980, Blood.

[69]  M. Minden,et al.  Self-renewal in culture of proliferative blast progenitor cells in acute myeloblastic leukemia. , 1979, Blood.