Peripheral blood stem cells collection by apheresis in very low-weight children with malignant diseases-A single center experience.

BACKGROUND: Hematopoietic stem cell transplantation is used in the treatment of children with malignant and non-malignant diseases. However, apheresis of peripheral blood stem cells (PBSC) represents a challenge in children below 10 kg. METHODS: A retrospective trial was conducted in the Cellular Therapy Department of Portuguese Oncology Institute of Porto. INCLUSION CRITERIA: children with body weight inferior to 10 kg who underwent autologous PBSC apheresis until 2021. Demographic and clinical data were collected and our institutional protocol was described. COBE Spectra apheresis system (TerumoBCT, Lakewood, Colorado) until 2012 and then Spectra Optia (TerumoBCT) were used. RESULTS: Sixteen leukocytaphereses were performed in 13 patients-nine females (69%). Mean age and weight were 13.31 months (±5.26) and 8.31 kg (±1.17), respectively. The initial CD34+ cells/µL in peripheral blood was 70.8 (±61.9). A central venous catheter (CVC) was exclusively used in all but one patient, in whom a peripheral vein was also required. In 10 procedures, both heparin and anticoagulant citrate dextrose solution-formula A (ACD-A) were used; in the remaining ones, only ACD-A was employed. The median duration of each procedure was 168 minutes (±45) and 2.96 blood volumes (±0.31) were processed. Median CD34+ cells yield per leukocytapheresis was 6.52 × 106 /kg (±9.87 × 106 ). CD34+ cell extraction rates were not significantly different between the two apheresis systems. Platelet and magnesium levels were significantly lower after collection (P < .001 and P = .009, respectively). CONCLUSIONS: Although recommendations are lacking, we have successfully and safely performed leukocytaphereses in children with body weight below 10 kg. The authors believe that a permanent and dynamic comprehensive evaluation of each child is paramount for attaining good results.

Therapeutic Leukapheresis: Experience of a Single Oncologic Centre.

Introduction: Leukostasis refers to clinical symptoms caused by hyperleukocytosis seen in some haematological diseases such as leukaemia. Cytoreduction can be achieved by therapeutic leukapheresis. The aim of this study was to retrospectively analyse the procedures performed in our Centre and to evaluate their efficacy and safety. Methods: This was a retrospective study of all the therapeutic leukapheresis procedures carried out in our Centre between January 1998 and December 2020. The sample collection was obtained through the review of the clinical files of the respective patients. Statistical analysis was performed using the software R v.4.0.1. A total of 54 therapeutic leukapheresis procedures were performed in 31 patients in our Centre. Results: After these procedures clinical improvement was observed in 16 patients and we verify that there was a significant difference in survival between the group that improved and the group that maintained the same clinical condition or worsened. The lack of immediate clinical improvement was a sign of a poor prognosis. Laboratory efficacy occurred in 16 patients who had a reduction in white blood cell count, with a 39.1% reduction after 24 h, and did not succeed in 15 patients, who had no reduction. However, in this case there is no significant difference in survival between the two groups. There was some complication in 53.9% of the procedures, with hypocalcaemia being the most frequent, which was observed in 22 procedures. Only 4 patients experienced serious side effects but these adverse reactions cannot be attributed to the procedures carried out. The overall survival rate 6 months after this treatment was 51.6%. Conclusion: Despite the reduced number of patients, we conclude that therapeutic leukapheresis is a safe and effective option that may still have a therapeutic role in some cases.

Allogeneic stem cell transplant in relapsed/ refractory Hodgkin lymphoma: A 21 years' experience.

Background: Allogenic stem cell transplant (alloSCT) has been used for several decades as a salvage strategy for relapsed/ refractory Hodgkin lymphoma (R/R HL), being a durable disease control method for some patients. Methods: A unicenter retrospective analysis was performed about alloSCT in R/R HL along 21 years. A survival analysis was made in search for prognostic factors with impact in overall survival (OS)/progression free survival (PFS). Results: Thirty-five patients were reviewed: median age 30years [17-46], 57.1% males, 82.9% had an esclero-nodular HL, 54.3% were in stage II of disease, and 42.9% achieved a complete response before the alloSCT. The donor type was matched-related in 54.3% and the stem cell source was peripheral blood in 97.1% of the grafts. All patients did a reduced intensity conditioning regimen. The overall response rate was 85.7% (complete in 68.6%, partial in 17.1%). Acute graft versus host disease grade II-IVwas seen in 45.7%. Transplant related mortality at day 360 was 17.9%. The median OS was 61 months (95% confidente interval: 33.6-88.3). The median PFS was 1Omonths (95% confidente interval: 3.1-16.9). Patients with >3Oyears at the alloSCT time and a previous autologous SCT showed better OS/PFS in the univariate analysis; having a matched donor and absence of infections along the alloSCT also improved PFS. Conclusions: AlloSCT is a feasible procedure in patients with R/R HL, being able to stabilize the disease in a large number of patients. However, it has a relevant toxicity in patients highly pre-treated.

Aggressive Neuroblastoma in a Pediatric Patient with Severe Hemophilia A.

Despite the extensive information regarding hemophilia's hemorrhagic complications, the literature on cancer in hemophilia is scarce, especially in pediatric patients. Many uncertainties remain concerning diagnosis and workup. We report a rare case of two severe diseases (neuroblastoma and hemophilia A (HA)) concomitantly present in the same pediatric patient. We highlight that the diagnosis of severe HA did not have a negative impact on the patient's oncologic course. This case also illustrates the significance of the cooperation among different specialties and hospitals when caring for the same patient.

Should we use extracorporeal photopheresis more often? Evidence from graft-versus-host disease patients monitored with Treg as a biomarker.

BACKGROUND: Chronic graft-versus-host disease (cGvHD) is a major complication after allogeneic hematopoietic cell transplantation. Extracorporeal photopheresis (ECP) is an immunotherapy treatment for cGvHD, although suitable response biomarkers are lacking. MATERIALS & METHODS: We analyzed data from six cGvHD patients undergoing ECP at a reference center from 826 to 2866 days. Circulating Tregs were enumerated, patient's clinical evolution, immunosuppression dose and adverse events (AEs) registered. RESULTS: We observed an increase in Tregs, a decrease in immunosuppression dosage and symptoms improvement. Mild AEs occurred at a very low rate. CONCLUSION: In these patients, the improvement of cGvHD, with low AEs, confirms a place for ECP as treatment. Improvements were accompanied by an increase in circulating Tregs, suggesting their role as a biomarker.

Stem cells out of the bag: characterization of ex vivo expanded mesenchymal stromal cells for possible clinical use.

AIM: Mesenchymal stromal cells (MSC) are a promising tool for cellular therapy and regenerative medicine. One major difficulty in establishing a MSC expansion protocol is the large volume of bone marrow (BM) required. We studied whether cells trapped within a collection bag and filter system could be considered as a source of MSC. RESULTS: From the 20 BM collection bag and filter systems, we recovered an average of 1.68 × 108 mononuclear cells, which is the equivalent to 60 ml of filtered BM. Mononuclear cells were expanded ex vivo to 17 × 106 MSC, with purity shown by a CD44+, CD105+, CD90+ and CD73+ immunophenotype, a reduction of 20% proliferating cells in a mixed lymphocyte reaction and also the ability of adipocyte differentiation. CONCLUSION: Long-term MSC cultures were established from the usually discarded BM collection bag and filter, maintaining an appropriate phenotype and function, being suitable for both investigation and clinical settings.

Monitoring the extracorporeal photopheresis by immunophenotyping in a Sézary syndrome patient.

The extracorporeal photopheresis is indicated as immunomodulatory treatment in primary cutaneous T-cell lymphoma. There are no guidelines about the most suitable immunophenotypic panel to monitor and correlate clinical evolution and laboratory parameters. In this study we characterize a patient with Sézary syndrome stage III, who performed 98 sessions of photopheresis; in the last 30, the expression of T-cells in peripheral blood was evaluated. The patient had exfoliative dermatitis (mainly in the lower limbs) and several analytical disorders: anemia, neutropenia and lymphocytosis; abnormal number of effector T-cells and a CD4/CD8 ratio higher than 10; atypical cells (CD7, CD26) with aberrant phenotype (characteristic of malignant T-cell clones); and regulatory T-cells indicating an immunotolerance state. The clinical evolution verified can be related with the therapeutic scheme adopted. We propose a multiparameter flow cytometry approach to monitor patients with Sézary syndrome that realize photopheresis, including the aberrant cases.

Outcomes of allogeneic stem cell transplantation among patients with acute myeloid leukemia presenting active disease: Experience of a single European Comprehensive Cancer Center.

INTRODUCTION:: Allogeneic hematopoietic stem cell transplantation (ASCT) representes a potentially curative approach for patients with relapsed or refractory acute myeloid leukemia (AML). We report the outcome of relapsed/refractory AML patients treated with ASCT. METHOD:: A retrospective cohort from 1994 to 2013 that included 61 patients with diagnosis of relapsed/refractory AML. Outcomes of interest were transplant-related mortality (TRM), incidence of acute and chronic graft-versus-host disease (GVHD), relapse incidence, progression-free survival (PFS) and overall survival (OS). Statistical significance was set at p<0.05. RESULTS:: The median age was 61 years (range 1 to 65). The cumulative incidence of 90 days, 1 year, and 3 years TRM were 60%, 26.7%, and 13.3%, respectively (p<0.001). The incidence of relapse was 21.7% at 1 year, 13% at 3 years, and 8.7% at 5 years. Median OS was estimated to be 8 months (95CI 3.266-12.734) and median PFS, 3 months (95CI 1.835-4.165). CONCLUSION:: In our cohort, TRM in first years after ASCT remains considerable, but ASCT in this setting seems to be a good choice for AML patients with active disease. However, novel approaches are needed to reduce TRM and relapse in this set of patients.

Outcomes of allogeneic stem cell transplantation among patients with acute myeloid leukemia presenting active disease: Experience of a single European Comprehensive Cancer Center / Resultados do transplante alogênico de células-tronco em doentes com leucemia mieloide aguda com doença ativa: experiência de um único Centro Oncológico Europeu

Summary Introduction: Allogeneic hematopoietic stem cell transplantation (ASCT) representes a potentially curative approach for patients with relapsed or refractory acute myeloid leukemia (AML). We report the outcome of relapsed/refractory AML patients treated with ASCT. Method: A retrospective cohort from 1994 to 2013 that included 61 patients with diagnosis of relapsed/refractory AML. Outcomes of interest were transplant-related mortality (TRM), incidence of acute and chronic graft-versus-host disease (GVHD), relapse incidence, progression-free survival (PFS) and overall survival (OS). Statistical significance was set at p<0.05. Results: The median age was 61 years (range 1 to 65). The cumulative incidence of 90 days, 1 year, and 3 years TRM were 60%, 26.7%, and 13.3%, respectively (p<0.001). The incidence of relapse was 21.7% at 1 year, 13% at 3 years, and 8.7% at 5 years. Median OS was estimated to be 8 months (95CI 3.266-12.734) and median PFS, 3 months (95CI 1.835-4.165). Conclusion: In our cohort, TRM in first years after ASCT remains considerable, but ASCT in this setting seems to be a good choice for AML patients with active disease. However, novel approaches are needed to reduce TRM and relapse in this set of patients.

Resumo Introdução: o transplante alogênico de células-tronco hematopoiéticas (TCTH-alo) representa uma abordagem potencialmente curativa para pacientes com leucemia mieloide aguda (LMA) recorrente ou refratária. Nosso trabalho apresenta o resultado de pacientes com recaída ou doença refratária tratados com TCTH-alo. Método: coorte retrospectiva incluindo 61 pacientes de 1994 a 2013 com diagnóstico de recidiva/LMA refratária. Os desfechos de interesse foram mortalidade relacionada ao transplante (MRT), incidência da doença aguda e crônica do enxerto contra hospedeiro (DECH), incidência de recaídas, sobrevida livre de progressão (PFS - progression-free survival) e sobrevida global (SG). A significância estatística foi considerada para p<0,05. Resultados: a média de idade foi de 61 anos (variação de 1 a 65). A incidência cumulativa de 90 dias, 1 ano e 3 anos de MRT foram de 60%, 26,7% e 13,3%, respectivamente (p<0,001). A incidência de recaída foi de 21,7% em 1 ano, 13% em 3 anos e 8,7% em 5 anos. A SG mediana foi estimada em 8 meses (IC 95% 3,266-12,734) e a mediana de PFS, em 3 meses (IC 95% 1,835-4,165). Conclusão: em nossa coorte, MRT no primeiro ano após o transplante permanece considerável, mas TCTH-alo nesse cenário parece ser uma boa opção para pacientes com LMA ativa. No entanto, novas abordagens são necessárias para reduzir MRT e recaída nesse conjunto de pacientes.

Twenty Years of Autologous Stem Cell Transplantation in Diffuse Large B-Cell Lymphoma: A Single Portuguese Center Experience.

INTRODUCTION: Diffuse large B-cell lymphoma can be cured in 60% - 70% of patients. Autologous stem cell transplantation is the standard treatment for relapsed disease. This high-intensity treatment after first complete remission in patients with high International Prognostic Index remains controversial and was performed in our department during some years. MATERIAL AND METHODS: Retrospective study, review of clinical records. RESULTS: This study evaluates the outcome of 113 patients transplanted between 1992 and 2012. Considering status before transplantation patients were divided in groups: a) first complete remission after 1 line of chemotherapy (n = 64); b) first complete remission after ≥ two chemotherapy lines (n = 15); c) second complete remission (n = 15); d) more advanced diseased (n = 19). Chemotherapy used in first line therapy was mainly R-CHOP (n = 71) and CHOP (n = 28). The median follow-up of patients still alive was 34 months (1 - 221). At five years, overall survival was 73% (± 5) and disease free survival was 75% (± 5). CONCLUSION: Conventional chemotherapy followed by autologous stem cell transplant is a safe and efficient option for selected patients. In our series 70% high-risk patients were free from disease with this strategy.

Introdução: O linfoma não Hodgkin difuso de grandes células B pode ser curado em 60% - 70% dos doentes. O transplante autológo de progenitores hematopoiéticos é o tratamento de intenção curativa standard à recidiva. Este tratamento intensivo após primeira remissão num grupo selecionado de doentes de alto risco é controverso e fez parte da estratégia do nosso Serviço durante alguns anos. Material e Métodos: Estudo retrospectivo, consulta do processo clínico. Resultados: Este estudo analisa o outcome de 113 doentes transplantados entre 1992 e 2012. Formaram-se quatro grupos com base no status pré-transplante: a) primeira remissão completa após 1 ciclo de quimioterapia (n = 64); b) segunda remissão completa após ≥ duas linhas de quimioterapia (n = 15); c) segunda remissão completa (n = 15); d) doença mais avançada (n = 19). O protocolo de quimioterapia de primeira linha mais utilizado foi R-CHOP (n = 71) e CHOP (n = 28). O seguimento mediano foi de 34 meses (1 - 221). Aos cinco anos a sobrevivência global foi de 73% (± 5) e a sobrevivência livre de progressão 75% (± 5). Conclusão: A imunoquimioterapia convencional seguida de transplante autólogo é uma opção segura e eficaz no tratamento de casos selecionados de linfoma difuso de grandes células B. Na nossa casuística cerca de 70% dos doentes de alto risco atingiram remissões duráveis com esta estratégia terapêutica.

Hematopoietic progenitor cells collection in pediatric patients with brain tumor.

AIM: To analyze the efficacy and safety of hematopoietic progenitor cells (HPC) collections by leukapheresis in pediatric patients with brain tumors. RESULTS: Between 2003 and 2014, we collected HPC from 19 children (12 boys/7 girls), median age at the diagnosis of 5 years old (<1-15 years old) and weight of 16.8 Kg (6.7-42). Diagnoses were Medulloblastoma (n = 10), Primitive Neuroectodermal Tumor (n = 5), Atypical Teratoid Rhabdoid Tumor (n = 3) and Secreting Germ Cell Tumor (n = 1). All patients performed leukapheresis by a central venous catheter, at the fifth day of mobilization with G-CSF (median dose 11.7 µg/Kg/day), 9 of them with COBE® Spectra and 10 with Spectra Optia®. The anticoagulant used was ACD-A, ratio of 14:1 or ACD-A plus heparin, ratio 25:1. The tubing set was primed with a compatible, irradiated, leukodepleted and hematocrit adjusted packed red cells for all children <30 kg (n = 17). A median of 1.5 (1-3) leukapheresis per patient was performed with an average of 3 (1.5-5.4) blood volumes processed; 3 children did a second mobilization and one additional leukapheresis. The median number of CD34+ cells collected was 4.6 × 10(6) /Kg (0.18-22.6) of patient body weight; 12 children collected for a tandem transplant. The median time between cell collection and infusion was 3 (0.6-9.1) months. CONCLUSIONS: HPC collection in children is an efficient and well tolerated technique, performed as an outpatient procedure. With the new mobilization schemes and leukapheresis technology, we can collect a high number of HPC allowing pediatric oncologist to establish more aggressive chemotherapy protocols hoping to improve patient outcome.