Molecular detection of c-mpl thrombopoietin receptor gene expression in chronic myeloproliferative disorders

Background-Chronic myeloproliferative disorders (CMPD) originate from a pluripotent haematopoietic progenitor cell but show a marked degree of heterogeneity, especially between Philadelphia chromosome positive and negative disease entities. Abnormal megakaryopoiesis is a frequent finding in CMPD, often associated with thrombocythaemic cell counts. Recent experimental data have suggested that the c-Mp1 thrombopoietin receptor, together with its ligand thrombopoietin, are not only the major physiological regulators of megakaryopoiesis and platelet production, but also play a crucial role in chronic myeloproliferation. Methods-A total of 18 peripheral blood mononuclear cell samples obtained from patients with CMPD (chronic myelocytic leukaemia (CML), n=10; polycythaemia vera (PV), n=6; and primary thrombocythaemia (PTH), n=2) were analysed for c-mp1 mRNA using the reverse transcriptase polymerase chain reaction (RT-PCR). In another 20 patients (CML, n=10; chronic megakaryocytic granulocytic myelosis (CMGM), n=3; PV, n=3; PTH, n=4), we compared the number of haematopoietic progenitors expressing c-Mp1, as characterised by coexpression with the CD34 antigen, in the bone marrow using double immunofluorescence staining. Results-c-mp1 mRNA was detected in all samples from patients with CML analysed, whereas only two of six PV and one of two PTH samples were positive (p less than or equal to 0.008; chi(2) test). Expression of the c-mp1 receptor gene was absent in healthy subjects used as controls. Similarly, an increase of c-Mp1 expressing CD34 positive haematopoietic cells was detected in seven of 10 bone marrow aspirates obtained from patients with CML. Increased numbers of c-Mp1 positive CD34 positive cells were found in only one of four patients with PTH, whereas in PV and CMGM the numbers of c-Mp1 positive CD34 positive cells did not exceed normal values, despite thrombocythaemic cell counts. Conclusions-These data confirm recent findings showing an impaired expression of the c-mp1 thrombopoietin receptor gene in Philadelphia chromosome negative CMPD when compared with patients with Philadelphia chromosome positive CML. The relevance of this observation to the functional and morphological characteristics of abnormal megakaryopoiesis remains unclear. Thrombocythaemic cell counts and a mature phenotype in megakaryocytes occur frequently in Philadelphia chromosome negative CMPD but require an intact c-Mp1 receptor under physiological conditions. Therefore, further studies are warranted to elucidate the mechanisms contributing to megakaryopoiesis in CMPD disease entities with decreased c-mp1 gene expression.