Gene expression profiling with principal component analysis depicts the biological continuum from essential thrombocythemia over polycythemia vera to myelofibrosis.

The recent discovery of the Janus activating kinase 2 V617F mutation in most patients with polycythemia vera (PV) and half of those with essential thrombocythemia (ET) and primary myelofibrosis (PMF) has favored the hypothesis of a biological continuum from ET over PV to PMF. We performed gene expression profiling of whole blood from control subjects (n = 21) and patients with ET (n = 19), PV (n = 41), and PMF (n = 9) using DNA microarrays. Applying an unsupervised method, principal component analysis, to search for patterns in the data, we demonstrated a separation of the four groups with biological relevant overlaps between the different entities. Moreover, the analysis separates Janus activating kinase 2-negative ET patients from Janus activating kinase 2-positive ET patients. Functional annotation analysis demonstrates that clusters of gene ontology terms related to inflammation, immune system, apoptosis, RNA metabolism, and secretory system were the most significantly deregulated terms in the three different disease groups. Our results yield further support for the hypothesis of a biological continuum originating from ET over PV to PMF. Functional analysis suggests an important implication of these gene ontology clusters in the pathogenesis of these neoplasms and in disease evolution from ET over PV to PMF.

[1]  T. Barbui,et al.  Leukocyte-platelet interaction in patients with essential thrombocythemia and polycythemia vera. , 2005, Experimental hematology.

[2]  J. Venables.  Aberrant and Alternative Splicing in Cancer , 2004, Cancer Research.

[3]  M. Jensen,et al.  Increased circulating platelet–leukocyte aggregates in myeloproliferative disorders is correlated to previous thrombosis, platelet activation and platelet count , 2001, European journal of haematology.

[4]  C. Richard,et al.  Expression of Bcl-x in erythroid precursors from patients with polycythemia vera. , 1998, The New England journal of medicine.

[5]  M. Björkholm,et al.  Autoimmunity and the risk of myeloproliferative neoplasms , 2010, Haematologica.

[6]  R. Hoffman,et al.  The renaissance of interferon therapy for the treatment of myeloid malignancies. , 2011, Blood.

[7]  W. Dameshek.  Editorial: Some Speculations on the Myeloproliferative Syndromes , 1951 .

[8]  T. Barbui,et al.  Inflammation and thrombosis in essential thrombocythemia and polycythemia vera: different role of C-reactive protein and pentraxin 3 , 2011, Haematologica.

[9]  R. Hoffman,et al.  Treatment with the Bcl-xL inhibitor ABT-737 in combination with interferon α specifically targets JAK2V617F-positive polycythemia vera hematopoietic progenitor cells. , 2010, Blood.

[10]  T. Barbui,et al.  V617F JAK-2 mutation in patients with essential thrombocythemia: relation to platelet, granulocyte, and plasma hemostatic and inflammatory molecules. , 2007, Experimental hematology.

[11]  R. Tiedt,et al.  Ratio of mutant JAK2-V617F to wild-type Jak2 determines the MPD phenotypes in transgenic mice. , 2007, Blood.

[12]  A. Kornblihtt,et al.  The connection between splicing and cancer , 2006, Journal of Cell Science.

[13]  Brad T. Sherman,et al.  Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists , 2008, Nucleic acids research.

[14]  Brad T. Sherman,et al.  Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources , 2008, Nature Protocols.

[15]  M. Cazzola,et al.  Altered gene expression in myeloproliferative disorders correlates with activation of signaling by the V617F mutation of Jak2. , 2005, Blood.

[16]  R. Levine,et al.  Expression of Jak2V617F causes a polycythemia vera-like disease with associated myelofibrosis in a murine bone marrow transplant model. , 2006, Blood.

[17]  M. Konopleva,et al.  Pegylated interferon alfa-2a yields high rates of hematologic and molecular response in patients with advanced essential thrombocythemia and polycythemia vera. , 2009, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[18]  T. Barbui,et al.  Survival and disease progression in essential thrombocythemia are significantly influenced by accurate morphologic diagnosis: an international study. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[19]  M. Jensen,et al.  Increased gene expression of histone deacetylases in patients with Philadelphia-negative chronic myeloproliferative neoplasms , 2010, Leukemia & lymphoma.

[20]  R. Zini,et al.  Molecular profile of CD34+ stem/progenitor cells according to JAK2V617F mutation status in essential thrombocythemia , 2009, Leukemia.

[21]  L. Arenillas,et al.  Gene expression profiling distinguishes JAK2V617F-negative from JAK2V617F-positive patients in essential thrombocythemia , 2008, Leukemia.

[22]  M. Møller,et al.  Minimal residual disease and normalization of the bone marrow after long-term treatment with alpha-interferon2b in polycythemia vera. A report on molecular response patterns in seven patients in sustained complete hematological remission , 2009, Hematology.

[23]  J. Thiele,et al.  A critical reappraisal of the WHO classification of the chronic myeloproliferative disorders , 2006, Leukemia & lymphoma.

[24]  W. Vainchenker,et al.  JAK2 stimulates homologous recombination and genetic instability: potential implication in the heterogeneity of myeloproliferative disorders. , 2008, Blood.

[25]  C. Langford,et al.  Gene expression profiling in polycythemia vera using cDNA microarray technology. , 2003, Cancer research.

[26]  D. Tenen,et al.  Apoptosis induced by JAK2 inhibition is mediated by Bim and enhanced by the BH3 mimetic ABT-737 in JAK2 mutant human erythroid cells. , 2010, Blood.

[27]  M. Jensen,et al.  High expression of carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 6 and 8 in primary myelofibrosis. , 2011, Leukemia research.

[28]  J. Fernández-Luna.  Apoptosis and polycythemia vera. , 1999, Current opinion in hematology.

[29]  W. Vainchenker,et al.  FLT3-mediated p38-MAPK activation participates in the control of megakaryopoiesis in primary myelofibrosis. , 2007, Cancer research.

[30]  Francesco Mannelli,et al.  Molecular Profiling of CD34+ Cells in Idiopathic Myelofibrosis Identifies a Set of Disease‐Associated Genes and Reveals the Clinical Significance of Wilms' Tumor Gene 1 (WT1) , 2007, Stem cells.

[31]  A. Tefferi,et al.  Novel mutations and their functional and clinical relevance in myeloproliferative neoplasms: JAK2, MPL, TET2, ASXL1, CBL, IDH and IKZF1 , 2010, Leukemia.

[32]  Cheng Li,et al.  Model-based analysis of oligonucleotide arrays: model validation, design issues and standard error application , 2001, Genome Biology.

[33]  W. Hofmann,et al.  Detection of Aberrant Gene Expression in CD34+ Hematopoietic Stem Cells from Patients with Agnogenic Myeloid Metaplasia Using Oligonucleotide Microarrays , 2005, Stem cells.

[34]  M. Jensen,et al.  Increased platelet activation and abnormal membrane glycoprotein content and redistribution in myeloproliferative disorders , 2000, British journal of haematology.

[35]  A. Tefferi.  Pathogenesis of myelofibrosis with myeloid metaplasia. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[36]  C. Ghigna,et al.  Alternative Splicing and Tumor Progression , 2008, Current genomics.

[37]  P. Guglielmelli,et al.  A quantitative assay for JAK2V617F mutation in myeloproliferative disorders by ARMS-PCR and capillary electrophoresis , 2006, Leukemia.

[38]  H. Hasselbalch,et al.  The JAK2 V617F mutation involves B‐ and T‐lymphocyte lineages in a subgroup of patients with Philadelphia‐chromosome negative chronic myeloproliferative disorders , 2007, British journal of haematology.

[39]  M. Monaghan,et al.  Class prediction models of thrombocytosis using genetic biomarkers. , 2010, Blood.

[40]  S. Chevret,et al.  Pegylated interferon-alfa-2a induces complete hematologic and molecular responses with low toxicity in polycythemia vera. , 2008, Blood.

[41]  A. Quintás-Cardama,et al.  New JAK2 inhibitors for myeloproliferative neoplasms , 2011, Expert opinion on investigational drugs.

[42]  R. Hoffman,et al.  Interferon-alpha targets JAK2V617F-positive hematopoietic progenitor cells and acts through the p38 MAPK pathway. , 2010, Experimental hematology.

[43]  C. Bloomfield,et al.  The 2008 revision of the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia: rationale and important changes. , 2009, Blood.

[44]  Mads Thomassen,et al.  Whole‐blood transcriptional profiling of interferon‐inducible genes identifies highly upregulated IFI27 in primary myelofibrosis , 2011, European journal of haematology.

[45]  Yi Li,et al.  Higher plant glycosyltransferases , 2001, Genome Biology.

[46]  Peter Harrison,et al.  Platelet a-granules , 1993 .

[47]  P. Campbell,et al.  The myeloproliferative disorders. , 2006, The New England journal of medicine.

[48]  M. Møller,et al.  The JAK2 V617F allele burden in essential thrombocythemia, polycythemia vera and primary myelofibrosis – impact on disease phenotype , 2007, European journal of haematology.

[49]  D. Gilliland,et al.  Transcriptional Profiling of Polycythemia Vera Identifies Gene Expression Patterns Both Dependent and Independent from the Action of JAK2V617F , 2010, Clinical Cancer Research.

[50]  S. Lewis,et al.  Transitional Myeloproliferative Disorder , 1979, British journal of haematology.

[51]  Peter J Campbell,et al.  Bone marrow pathology in essential thrombocythemia: interobserver reliability and utility for identifying disease subtypes. , 2008, Blood.