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9. Molecular purging of multiple myeloma cells by ex-vivo culture and retroviral transduction of mobilized-blood CD34+ cells

Author

Corneo Gianmarco, Pogliani Enrico, Monari Marta, Vai Sergio, Voena Claudia, Dando Jonathan, Ficara Francesca, Cergnul Massimiliano, Birolo Roberto, Scaramuzza Samantha, Deola Sara, Peccatori Jacopo, Selleri Silvia, Bordignon Claudio, Roncarolo Maria, Aiuti Alessandro, and Bregni Marco

Subjects
Medicine
Abstract

Abstract Background Tumor cell contamination of the apheresis in multiple myeloma is likely to affect disease-free and overall survival after autografting. Objective To purge myeloma aphereses from tumor contaminants with a novel culture-based purging method. Methods We cultured myeloma-positive CD34+ PB samples in conditions that retained multipotency of hematopoietic stem cells, but were unfavourable to survival of plasma cells. Moreover, we exploited the resistance of myeloma plasma cells to retroviral transduction by targeting the hematopoietic CD34+ cell population with a retroviral vector carrying a selectable marker (the truncated form of the human receptor for nerve growth factor, ΔNGFR). We performed therefore a further myeloma purging step by selecting the transduced cells at the end of the culture. Results Overall recovery of CD34+ cells after culture was 128.5%; ΔNGFR transduction rate was 28.8% for CD34+ cells and 0% for CD138-selected primary myeloma cells, respectively. Recovery of CD34+ cells after ΔNGFR selection was 22.3%. By patient-specific Ig-gene rearrangements, we assessed a decrease of 0.7–1.4 logs in tumor load after the CD34+ cell selection, and up to 2.3 logs after culture and ΔNGFR selection. Conclusion We conclude that ex-vivo culture and retroviral-mediated transduction of myeloma leukaphereses provide an efficient tumor cell purging.

Published
2007
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15. Role of [Alpha]1 Acid Glycoprotein in the In Vivo Resistance of Human BCR-[ABL.sup.+] Leukemic Cells to the Abl Inhibitor STI571

Author

Gambacorti-Passerini, Carlo, Barni, Rossella, le Coutre, Philipp, Zucchetti, Massimo, Cabrita, Goncalo, Cleris, Loredana, Rossi, Francesca, Gianazza, Elisabetta, Brueggen, Josef, Cozens, Robert, Pioltelli, Pietro, Pogliani, Enrico, Corneo, Gianmarco, Formelli, Franca, and D'Incalci, Maurizio

Subjects
Myelocytic leukemia -- Models, Glycoproteins -- Physiological aspects, Health
Abstract

Background: Chronic myeloid leukemia is caused by a chromosomal translocation that results in an oncogenic fusion protein, Bcr-Abl. Bcr-Abl is a tyrosine kinase whose activity is inhibited by the antineoplastic drug STI571. This drug can cure mice given an injection of human leukemic cells, but treatment ultimately fails in animals that have large tumors when treatment is initiated. We created a mouse model to explore the mechanism of resistance in vivo. Methods: Nude mice were injected with KU812 Bcr-[Abl.sup.+] human leukemic cells. After 1 day (no evident tumors), 8 days, or 15 days (tumors [is greater than] 1 g), mice were treated with STI571 (160 mg/kg every 8 hours). Cells recovered from relapsing animals were used for in vitro experiments. Statistical tests were two-sided. Results: Tumors regressed initially in all STI571-treated mice, but all mice treated 15 days after injection of tumor cells eventually relapsed. Relapsed animals did not respond to further STI571 treatment, and their Bcr-Abl kinase activity in vivo was not inhibited by STI571, despite high plasma concentrations of the drug. However, tumor cells from resistant animals were sensitive to STI571 in vitro, suggesting that a molecule in the plasma of relapsed animals may inactivate the drug. The plasma protein [Alpha]1 acid glycoprotein (AGP) bound STI571 at physiologic concentrations in vitro and blocked the ability of STI571 to inhibit Bcr-Abl kinase activity in a dose-dependent manner. Plasma AGP concentrations were strongly associated with tumor load. Erythromycin competed with STI571 for AGP binding. When animals bearing large tumors were treated with STI571 alone or with a combination of STI571 and erythromycin, greater tumor reductions and better long-term tumor-free survival (10 of 12 versus one of 13 at day 180; P [is less than] .001) were observed after the combination treatment. Conclusion: AGP in the plasma of relapsed animals binds to STI571, preventing this compound from inhibiting the Bcr/Abl tyrosine kinase. Molecules such as erythromycin that compete with STI571 for binding to AGP may enhance the therapeutic potential of this drug. [J Natl Cancer Inst 2000;92:1641-50]

Published
2000

17. Mechanisms of resistance to imatinib in chronic myeloid leukemia (CML) patients in chronic phase (CP) who achieved durable complete cytogenetic responses (CCR)

Author

PIAZZA, ROCCO GIOVANNI, MAGISTRONI, VERA, CAZZANIGA, GIOVANNI ITALO, BIONDI, ANDREA, POGLIANI, ENRICO MARIA, CORNEO, GIANMARCO, Tornaghi, L, Andreoni, F, Bungaro, S, Varella Garcia, M, GAMBACORTI PASSERINI, CARLO, Piazza, R, Magistroni, V, Tornaghi, L, Andreoni, F, Bungaro, S, Cazzaniga, G, Biondi, A, Pogliani, E, Varella Garcia, M, Corneo, G, and GAMBACORTI PASSERINI, C

Subjects
MED/15 - MALATTIE DEL SANGUE, Imatinib, Resistance, Complete Cytogenetic Response
Published
2003

18. Mechanisms of resistance to imatinib in chronic myeloid leukemia (CML) patients in chronic phase (CP) who achieved durable complete cytogenetic responses (CCR)

Author

Piazza, R, Magistroni, V, Tornaghi, L, Andreoni, F, Bungaro, S, Cazzaniga, G, Biondi, A, Pogliani, E, Varella Garcia, M, Corneo, G, GAMBACORTI PASSERINI, C, PIAZZA, ROCCO GIOVANNI, MAGISTRONI, VERA, CAZZANIGA, GIOVANNI ITALO, BIONDI, ANDREA, POGLIANI, ENRICO MARIA, CORNEO, GIANMARCO, GAMBACORTI PASSERINI, CARLO, Piazza, R, Magistroni, V, Tornaghi, L, Andreoni, F, Bungaro, S, Cazzaniga, G, Biondi, A, Pogliani, E, Varella Garcia, M, Corneo, G, GAMBACORTI PASSERINI, C, PIAZZA, ROCCO GIOVANNI, MAGISTRONI, VERA, CAZZANIGA, GIOVANNI ITALO, BIONDI, ANDREA, POGLIANI, ENRICO MARIA, CORNEO, GIANMARCO, and GAMBACORTI PASSERINI, CARLO

Published
2003

19. Corrigendum: In reply to 'Cardiotoxicity of the cancer therapeutic agent imatinib mesylate'

Author

Gambacorti-Passerini, Carlo, Tornaghi, Lucia, Franceschino, Anna, Piazza, Rocco, Corneo, Gianmarco, and Pogliani, Enrico

Abstract

Author(s): Carlo Gambacorti-Passerini; Lucia Tornaghi; Anna Franceschino; Rocco Piazza; Gianmarco Corneo; Enrico Pogliani Nature Medicine 13, 13-14 (2007); corrected after print 25 January 2007 In the version of the correspondence [...]

Published
2007
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20. Molecular purging of multiple myeloma cells by ex-vivo culture and retroviral transduction of mobilized-blood CD34+ cells

Author

Deola, Sara, primary, Scaramuzza, Samantha, additional, Birolo, Roberto Sciarretta, additional, Cergnul, Massimiliano, additional, Ficara, Francesca, additional, Dando, Jonathan, additional, Voena, Claudia, additional, Vai, Sergio, additional, Monari, Marta, additional, Pogliani, Enrico, additional, Corneo, Gianmarco, additional, Peccatori, Jacopo, additional, Selleri, Silvia, additional, Bordignon, Claudio, additional, Roncarolo, Maria Grazia, additional, Aiuti, Alessandro, additional, and Bregni, Marco, additional

Published
2007
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22. Risk-oriented postremission strategies in adult acute lymphoblastic leukemia: prospective confirmation of anthracycline activity in standard-risk class and role of hematopoietic stem cell transplants in high-risk groups

Author

Bassan, Renato, primary, Pogliani, Enrico, additional, Casula, Paolo, additional, Rossi, Giuseppe, additional, Fabris, Pietro, additional, Morandi, Sergio, additional, Lambertenghi-Deliliers, Giorgio, additional, Vespignani, Michele, additional, Lerede, Teresa, additional, Rambaldi, Alessandro, additional, Borleri, Gianmaria, additional, Spedini, Pierangelo, additional, Cortelezzi, Agostino, additional, Izzi, Teodosio, additional, Coser, Paolo, additional, Broccia, Giorgio, additional, Corneo, Gianmarco, additional, and Barbui, Tiziano, additional

Published
2001
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24. Granulocyte colony-Stimulating factor (G-CSF, filgrastim) after or during an intensive remission induction therapy for adult acute lymphoblastic leukaemia: Effects, role of patient pretreatment characteristics, and costs

Author

Bassan, Renato, primary, Lerede, Teresa, additional, Bona, Eros Di, additional, Rossi, Giuseppe, additional, Pogliani, Enrico, additional, Rambaldi, Alessandro, additional, Buelli, Maurizio, additional, Viero, Piera, additional, Rodeghiero, Francesco, additional, Izzi, Teodosio, additional, Corneo, Gianmarco, additional, and Barbui, Tiziano, additional

Published
1997
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29. Molecular purging of multiple myeloma cells by ex-vivo culture and retroviral transduction of mobilized-blood CD34+ cells.

Author

Deola, Sara, Scaramuzza, Samantha, Birolo, Roberto Sciarretta, Cergnul, Massimiliano, Ficara, Francesca, Dando, Jonathan, Voena, Claudia, Vai, Sergio, Monari, Marta, Pogliani, Enrico, Corneo, Gianmarco, Peccatori, Jacopo, Selleri, Silvia, Bordignon, Claudio, Roncarolo, Maria Grazia, Aiuti, Alessandro, and Bregni, Marco

Subjects
MULTIPLE myeloma, B cell lymphoma, CANCER cells, HEMATOPOIETIC stem cells, BONE marrow cells, CELL proliferation, CYTOKINES
Abstract

Background: Tumor cell contamination of the apheresis in multiple myeloma is likely to affect disease-free and overall survival after autografting. Objective: To purge myeloma aphereses from tumor contaminants with a novel culture-based purging method. Methods: We cultured myeloma-positive CD34+ PB samples in conditions that retained multipotency of hematopoietic stem cells, but were unfavourable to survival of plasma cells. Moreover, we exploited the resistance of myeloma plasma cells to retroviral transduction by targeting the hematopoietic CD34+ cell population with a retroviral vector carrying a selectable marker (the truncated form of the human receptor for nerve growth factor, ΔNGFR). We performed therefore a further myeloma purging step by selecting the transduced cells at the end of the culture. Results: Overall recovery of CD34+ cells after culture was 128.5%; ΔNGFR transduction rate was 28.8% for CD34+ cells and 0% for CD138-selected primary myeloma cells, respectively. Recovery of CD34+ cells after ΔNGFR selection was 22.3%. By patient-specific Ig-gene rearrangements, we assessed a decrease of 0.7-1.4 logs in tumor load after the CD34+ cell selection, and up to 2.3 logs after culture and ΔNGFR selection. Conclusion: We conclude that ex-vivo culture and retroviral-mediated transduction of myeloma leukaphereses provide an efficient tumor cell purging. [ABSTRACT FROM AUTHOR]

Published
2007
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34. Homology between Cellular Repeated Nucleotide Sequences and a Murine Leukemia Viral Genome1

Author

Ginelli, Enrico, Gianni, Alessandro M., and Corneo, Gianmarco

Abstract

Moloney murine leukemia virus c-DNA hybridizes mainly with cellular middle repeated sequences of NIH-Swiss mouse spleen DNA, fragmented to different lengths, denatured and renatured to an intermediate value of Cot, and fractionated in an Ag+-Cs2SO4preparative gradient suitable to separate unique, middle repeated and highly repeated DNA.

Published
1978
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36. Role of α1 Acid Glycoprotein in the In Vivo Resistance of Human BCR-ABL+ Leukemic Cells to the Abl Inhibitor STI571

Author

Gambacorti-Passerini, Carlo, Barni, Rossella, le Coutre, Philipp, Zucchetti, Massimo, Cabrita, Gonçalo, Cleris, Loredana, Rossi, Francesca, Gianazza, Elisabetta, Brueggen, Josef, Cozens, Robert, Pioltelli, Pietro, Pogliani, Enrico, Corneo, Gianmarco, Formelli, Franca, D'Incalci, Maurizio, Gambacorti-Passerini, Carlo, Barni, Rossella, le Coutre, Philipp, Zucchetti, Massimo, Cabrita, Gonçalo, Cleris, Loredana, Rossi, Francesca, Gianazza, Elisabetta, Brueggen, Josef, Cozens, Robert, Pioltelli, Pietro, Pogliani, Enrico, Corneo, Gianmarco, Formelli, Franca, and D'Incalci, Maurizio

Abstract

Background: Chronic myeloid leukemia is caused by a chromosomal translocation that results in an oncogenic fusion protein, Bcr-Abl. Bcr-Abl is a tyrosine kinase whose activity is inhibited by the antineoplastic drug STI571. This drug can cure mice given an injection of human leukemic cells, but treatment ultimately fails in animals that have large tumors when treatment is initiated. We created a mouse model to explore the mechanism of resistance in vivo. Methods: Nude mice were injected with KU812 Bcr-Abl+ human leukemic cells. After 1 day (no evident tumors), 8 days, or 15 days (tumors >1 g), mice were treated with STI571 (160 mg/kg every 8 hours). Cells recovered from relapsing animals were used for in vitro experiments. Statistical tests were two-sided. Results: Tumors regressed initially in all STI571-treated mice, but all mice treated 15 days after injection of tumor cells eventually relapsed. Relapsed animals did not respond to further STI571 treatment, and their Bcr-Abl kinase activity in vivo was not inhibited by STI571, despite high plasma concentrations of the drug. However, tumor cells from resistant animals were sensitive to STI571 in vitro, suggesting that a molecule in the plasma of relapsed animals may inactivate the drug. The plasma protein α1 acid glycoprotein (AGP) bound STI571 at physiologic concentrations in vitro and blocked the ability of STI571 to inhibit Bcr-Abl kinase activity in a dose-dependent manner. Plasma AGP concentrations were strongly associated with tumor load. Erythromycin competed with STI571 for AGP binding. When animals bearing large tumors were treated with STI571 alone or with a combination of STI571 and erythromycin, greater tumor reductions and better long-term tumor-free survival (10 of 12 versus one of 13 at day 180; P<.001) were observed after the combination treatment. Conclusion: AGP in the plasma of relapsed animals binds to STI571, preventing this compound from inhibiting the Bcr/Abl tyrosine kinase. Molecules such a

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