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

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

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Article number35
JournalJournal of Translational Medicine
Volume5
DOIs
Publication statusPublished - 12 Jul 2007
Externally publishedYes

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Purging
Multiple Myeloma
Tumors
Blood Cells
Nerve Growth Factor
Blood
Cells
Plasmas
Nerve Growth Factor Receptor
Recovery
Blood Component Removal
Stem cells
Contamination
Genes
Impurities
Leukapheresis
Neoplasms
Immunoglobulin Genes
Gene Rearrangement
Autologous Transplantation

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Molecular purging of multiple myeloma cells by ex-vivo culture and retroviral transduction of mobilized-blood CD34+ cells. / 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; Bregni, Marco.

In: Journal of Translational Medicine, Vol. 5, 35, 12.07.2007.

Research output: Contribution to journalArticle

Deola, S, Scaramuzza, S, Birolo, RS, Cergnul, M, Ficara, F, Dando, J, Voena, C, Vai, S, Monari, M, Pogliani, E, Corneo, G, Peccatori, J, Selleri, S, Bordignon, C, Roncarolo, MG, Aiuti, A & Bregni, M 2007, 'Molecular purging of multiple myeloma cells by ex-vivo culture and retroviral transduction of mobilized-blood CD34+ cells', Journal of Translational Medicine, vol. 5, 35. https://doi.org/10.1186/1479-5876-5-35
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 ; Bregni, Marco. / Molecular purging of multiple myeloma cells by ex-vivo culture and retroviral transduction of mobilized-blood CD34+ cells. In: Journal of Translational Medicine. 2007 ; Vol. 5.
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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.",
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AU - Deola, Sara

AU - Scaramuzza, Samantha

AU - Birolo, Roberto Sciarretta

AU - Cergnul, Massimiliano

AU - Ficara, Francesca

AU - Dando, Jonathan

AU - Voena, Claudia

AU - Vai, Sergio

AU - Monari, Marta

AU - Pogliani, Enrico

AU - Corneo, Gianmarco

AU - Peccatori, Jacopo

AU - Selleri, Silvia

AU - Bordignon, Claudio

AU - Roncarolo, Maria Grazia

AU - Aiuti, Alessandro

AU - Bregni, Marco

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N2 - 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.

AB - 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.

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