Correlation of the in vitro formation of endogenous erythroid and megakaryocytic circulating colonies with HUMARA clonality assay and PRV-1 expression in polycythemia vera and essential thrombocythemia

Florensa, L.; Zamora, L.; Besses, C.; Bellosillo, B.; Espinet, B.; Serrano, S.; Woessner, S.; Sole, F.

Blood 102(11): 337b

2003


ISSN/ISBN: 0006-4971
Accession: 034655546

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Abstract
Introduction: Polycythemia vera (PV) and essential thrombocythemia (ET) are clonal myeloproliferative disorders that are sometimes difficult to distinguish from secondary erythrocytosis and reactive thrombocytosis. In this context, some laboratory tests have been used for differentiating clonal from non clonal myeloid proliferations. The aim of the present study was to analyze the possible correlations among several diagnostic assays (in vitro culture, HUMARA clonality and expression of PRV-1) which provide myeloproliferative hallmarks to polycythemic and thrombocythemic disorders. Patients and methods: Patients: 28 females (range 30-86 years) affected of PV (n=11) and ET (n=17) were included in this study. All patients fulfilled the diagnosis criteria of PVSG. Methods: endogenous growth of BFU-E (without EPO) and CFU-MK (without LCM) were studied. XCIP by HUMARA-PCR was performed on DNA of purified granulocytes and CD3+ lymphocytes. Expression of PRV-1 and the housekeeping gene (GUS) from RNA of granulocytes was quantified by real time RT-PCR. Results: In PV patients, 11/11 (100%) showed endogenous BFU-E growth and 4/11 (36%) also had endogenous CFU-MK growth. HUMARA clonality was demonstrated in 5 out of 10 patients, the remaining patient was homozygous. PRV-1 overexpression was detected in 10/11 patients (91%). Regarding ET patients, 5/17 (29%) had endogenous BFU-E growth, 7/17 (41%) showed endogenous CFU-MK growth and 9/17 (53%) had endogenous BFU-E and/or CFU-MK growth. Clonal hematopoiesis was demonstrated in 6/14 patients (43%), in the other three patients the technique was non informative. PRV-1 overexpression was observed in 10 out of 17 patients (59%). In PV patients, 10/11 (91%) showed a simultaneous positive result between endogenous BFU-E and PRV-1 expression. The correlation among the three assays gave a concurrent positve result in two patients, but if the three assays were considered separately, we found a positive result in at least one of them in all PV. Concerning ET patients, from the 9 patients with endogenous growth (BFU-E and/or CFU-MK), 7 had overexpression of PRV-1 (78%). The correlation among the three assays gave a concurrent positve result in two patients, but if the three assays are considered separately, we found a positive result in at least one of them in 15 out of 17 ET (88%).