RESUMO
Autologous stem cell transplantation is used to rescue cancer patients from myelosuppression caused by high-dose chemotherapy. However, autologous grafts often contain tumor cells that can contribute directly to relapse. Current purging methods are useful when fewer than 1% tumor cells contaminate the bone marrow, and patients with tumor burdens of >1% are considered ineligible for chemotherapy that necessitates stem cell rescue. Using neuroblastoma (NB) as a model system, we developed a method that is effective even with tumor burdens of 10-25%. Mixtures of NB-1691 NB cells and CD34(+) hematopoietic cells purged by this method showed no evidence of viable tumor cells as assessed by clonogenic assays or reverse transcription-PCR for the NB cell markers tyrosine hydroxylase and N-MYC. The efficacy and lack of toxicity of the method were verified using in vivo mouse models. Severe combined immunodeficient mice that received purged cell preparations containing 10% NB-1691 cells survived without evidence of disease for the observation period (>1 year), whereas mice that received unpurged cells developed disseminated disease requiring euthanasia 73-96 days after injection of cells. No evidence of toxicity to the mice was detected by numerous laboratory values for bone marrow, liver, and kidney function, and no difference was seen in the ability of purged cell mixtures versus unmanipulated CD34(+) cells to reconstitute the marrow of non-obese diabetic severe combined immunodeficient mice. In a pilot study, marrow was obtained from eight patients who had >/=1% metastatic tumor burden. All eight samples were purged to the level of detection by reverse transcription-PCR (samples from seven patients) or clonogenic potential (sample from one patient), whichever assay was used. The described adenovirus/rabbit carboxylesterase/CPT-11 (irinotecan, 7-ethyl-10[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin) virus-directed enzyme prodrug method may be useful for patients whose tumor burdens exceed 1% at the time of stem cell harvest. Assessment of purging efficacy with additional samples from NB patients is ongoing.
Assuntos
Purging da Medula Óssea/métodos , Camptotecina/farmacocinética , Hidrolases de Éster Carboxílico/genética , Neuroblastoma/patologia , Pró-Fármacos/farmacocinética , Adenoviridae/genética , Animais , Medula Óssea/patologia , Purging da Medula Óssea/efeitos adversos , Camptotecina/análogos & derivados , Hidrolases de Éster Carboxílico/metabolismo , DNA Complementar/genética , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Transplante de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas/patologia , Humanos , Irinotecano , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Neuroblastoma/terapia , Projetos PilotoRESUMO
Relapse due to either residual host disease or reinfused tumor cells remains the principal cause of treatment failure after autologous stem cell transplantation. Although it is intuitively attractive to remove putative tumor cells from autologous grafts prior to transplant and more than 1000 articles have been written on the subject, there are only limited data suggesting that purging autografts has any favorable effect on relapses or disease-free survival. Certain purging techniques that remove substantial numbers of T cells or destroy progenitor cells may have adverse effects such as delayed hematopoietic or T cell reconstitution. There is a critical need for large, well-designed trials that specifically address the value of a particular purging technique on relapses and disease-free survival after autologous stem cell transplant.