Uuantimcatiun of bcr-abl ikanscripts by competitive polymerase chain reaction in leukapheresis-collected blood progenitor cells from early diagnosed patients with chronic myeloid leukemia

M. T. Corsetti, F. Frassoni, M. Podestà, E. Lerma, A. Dejana, L. Celesti, C. Parodi, F. Manca, A. M. Carella

Research output: Contribution to journalArticle

Abstract

Reduction of clonal genetically unstable cells is the main therapeutic goal in chronic myeloid leukemia (CML) as it is thought to bt necessary for reducing the rate of secondary genetic changes and, thereby, for postponing blast crisis. We have shown that Ph-negative blood progenitor cells (BPC) can be collected by leukapheresis during early recovery after chemoterapy-induced marrow aplasia and that these cells are able to restore normal polyclonal hemopoiesis after autografting. As we have also shown that the best clinical results are achieved in patients autografted with Ph-negative BPC, the degree of the tumor burden in BPC could correlate with a prolongation in survival. In order to estimate the level of contamination with residual BCR-ABL positive cells, leukapheresis-collected peripheral blood mononuclear cells (PBMC) have been monitored by competitive reverse transcriptase-polymerase chain reaction (RT-PCR) for Phchromosome originated BCR-ABL transcripts. The tested leukapheresis-collected PBMC were from four patients; 100% cytogenetically Ph-negative PBMC were recovered from three of them, while PBMC from the fourth patient showed to have degrees of cytogenetically detectable Ph-positive cells. In the three patients with Ph-negative PBMC, competitive RT-PCR showed that, in cells from the first leukapheresis, BCR-ABL transcripts were under the threshold of 104/jig RNA, while the number of BCR-ABL transcripts increased in successive leukaphereses from two patients and remained under the threshold in the third one. PBMC from the partially Ph-positive patient showed little variation in level of BCR-ABL transcripts as in all leukaphereses at least 3 × 104 transcripts/ng RNA were found. According to other previous observations, also these preliminary data seem to suggest that normal Ph-negative progenitor cells increase in peripheral blood earlier than leukemic Ph-positive ones and that the combination of cytogenetics and molecular methods may significantly improve the ability to monitor the tumor burden in leukapheresiscollected PBMC.

Original languageEnglish
Pages (from-to)869
Number of pages1
JournalExperimental Hematology
Volume25
Issue number8
Publication statusPublished - 1997

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Leukapheresis
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Blood Cells
Stem Cells
Polymerase Chain Reaction
Tumor Burden
Reverse Transcriptase Polymerase Chain Reaction
RNA
Blast Crisis
Autologous Transplantation
Cytogenetics
Bone Marrow

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

@article{8de28d35745b4243a19fdc63589a1332,
title = "Uuantimcatiun of bcr-abl ikanscripts by competitive polymerase chain reaction in leukapheresis-collected blood progenitor cells from early diagnosed patients with chronic myeloid leukemia",
abstract = "Reduction of clonal genetically unstable cells is the main therapeutic goal in chronic myeloid leukemia (CML) as it is thought to bt necessary for reducing the rate of secondary genetic changes and, thereby, for postponing blast crisis. We have shown that Ph-negative blood progenitor cells (BPC) can be collected by leukapheresis during early recovery after chemoterapy-induced marrow aplasia and that these cells are able to restore normal polyclonal hemopoiesis after autografting. As we have also shown that the best clinical results are achieved in patients autografted with Ph-negative BPC, the degree of the tumor burden in BPC could correlate with a prolongation in survival. In order to estimate the level of contamination with residual BCR-ABL positive cells, leukapheresis-collected peripheral blood mononuclear cells (PBMC) have been monitored by competitive reverse transcriptase-polymerase chain reaction (RT-PCR) for Phchromosome originated BCR-ABL transcripts. The tested leukapheresis-collected PBMC were from four patients; 100{\%} cytogenetically Ph-negative PBMC were recovered from three of them, while PBMC from the fourth patient showed to have degrees of cytogenetically detectable Ph-positive cells. In the three patients with Ph-negative PBMC, competitive RT-PCR showed that, in cells from the first leukapheresis, BCR-ABL transcripts were under the threshold of 104/jig RNA, while the number of BCR-ABL transcripts increased in successive leukaphereses from two patients and remained under the threshold in the third one. PBMC from the partially Ph-positive patient showed little variation in level of BCR-ABL transcripts as in all leukaphereses at least 3 × 104 transcripts/ng RNA were found. According to other previous observations, also these preliminary data seem to suggest that normal Ph-negative progenitor cells increase in peripheral blood earlier than leukemic Ph-positive ones and that the combination of cytogenetics and molecular methods may significantly improve the ability to monitor the tumor burden in leukapheresiscollected PBMC.",
author = "Corsetti, {M. T.} and F. Frassoni and M. Podest{\`a} and E. Lerma and A. Dejana and L. Celesti and C. Parodi and F. Manca and Carella, {A. M.}",
year = "1997",
language = "English",
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pages = "869",
journal = "Experimental Hematology",
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T1 - Uuantimcatiun of bcr-abl ikanscripts by competitive polymerase chain reaction in leukapheresis-collected blood progenitor cells from early diagnosed patients with chronic myeloid leukemia

AU - Corsetti, M. T.

AU - Frassoni, F.

AU - Podestà, M.

AU - Lerma, E.

AU - Dejana, A.

AU - Celesti, L.

AU - Parodi, C.

AU - Manca, F.

AU - Carella, A. M.

PY - 1997

Y1 - 1997

N2 - Reduction of clonal genetically unstable cells is the main therapeutic goal in chronic myeloid leukemia (CML) as it is thought to bt necessary for reducing the rate of secondary genetic changes and, thereby, for postponing blast crisis. We have shown that Ph-negative blood progenitor cells (BPC) can be collected by leukapheresis during early recovery after chemoterapy-induced marrow aplasia and that these cells are able to restore normal polyclonal hemopoiesis after autografting. As we have also shown that the best clinical results are achieved in patients autografted with Ph-negative BPC, the degree of the tumor burden in BPC could correlate with a prolongation in survival. In order to estimate the level of contamination with residual BCR-ABL positive cells, leukapheresis-collected peripheral blood mononuclear cells (PBMC) have been monitored by competitive reverse transcriptase-polymerase chain reaction (RT-PCR) for Phchromosome originated BCR-ABL transcripts. The tested leukapheresis-collected PBMC were from four patients; 100% cytogenetically Ph-negative PBMC were recovered from three of them, while PBMC from the fourth patient showed to have degrees of cytogenetically detectable Ph-positive cells. In the three patients with Ph-negative PBMC, competitive RT-PCR showed that, in cells from the first leukapheresis, BCR-ABL transcripts were under the threshold of 104/jig RNA, while the number of BCR-ABL transcripts increased in successive leukaphereses from two patients and remained under the threshold in the third one. PBMC from the partially Ph-positive patient showed little variation in level of BCR-ABL transcripts as in all leukaphereses at least 3 × 104 transcripts/ng RNA were found. According to other previous observations, also these preliminary data seem to suggest that normal Ph-negative progenitor cells increase in peripheral blood earlier than leukemic Ph-positive ones and that the combination of cytogenetics and molecular methods may significantly improve the ability to monitor the tumor burden in leukapheresiscollected PBMC.

AB - Reduction of clonal genetically unstable cells is the main therapeutic goal in chronic myeloid leukemia (CML) as it is thought to bt necessary for reducing the rate of secondary genetic changes and, thereby, for postponing blast crisis. We have shown that Ph-negative blood progenitor cells (BPC) can be collected by leukapheresis during early recovery after chemoterapy-induced marrow aplasia and that these cells are able to restore normal polyclonal hemopoiesis after autografting. As we have also shown that the best clinical results are achieved in patients autografted with Ph-negative BPC, the degree of the tumor burden in BPC could correlate with a prolongation in survival. In order to estimate the level of contamination with residual BCR-ABL positive cells, leukapheresis-collected peripheral blood mononuclear cells (PBMC) have been monitored by competitive reverse transcriptase-polymerase chain reaction (RT-PCR) for Phchromosome originated BCR-ABL transcripts. The tested leukapheresis-collected PBMC were from four patients; 100% cytogenetically Ph-negative PBMC were recovered from three of them, while PBMC from the fourth patient showed to have degrees of cytogenetically detectable Ph-positive cells. In the three patients with Ph-negative PBMC, competitive RT-PCR showed that, in cells from the first leukapheresis, BCR-ABL transcripts were under the threshold of 104/jig RNA, while the number of BCR-ABL transcripts increased in successive leukaphereses from two patients and remained under the threshold in the third one. PBMC from the partially Ph-positive patient showed little variation in level of BCR-ABL transcripts as in all leukaphereses at least 3 × 104 transcripts/ng RNA were found. According to other previous observations, also these preliminary data seem to suggest that normal Ph-negative progenitor cells increase in peripheral blood earlier than leukemic Ph-positive ones and that the combination of cytogenetics and molecular methods may significantly improve the ability to monitor the tumor burden in leukapheresiscollected PBMC.

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