RESUMEN
BACKGROUND: Nivestym, a biosimilar granulocyte colony-stimulating factor (G-CSF) to the originator filgrastim (Neupogen), is now being used for the mobilization of peripheral blood stem cells (PBSC) in allogeneic hematopoietic stem cell transplantation (allo-HSCT). We aim to compare the efficacy of Nivestym and Neupogen for PBSC mobilization in healthy allogeneic donors. METHODS: We conducted a retrospective single-center study including 541 adult allo-HSCT donors receiving Nivestym (January 2013-July 2020), or Neupogen (July 2020-June 2023) for donor PBSC mobilization. Bivariate analysis was conducted using SPSS version 28. Statistical significance was determined at a p-value <.05. RESULTS: Our study included 541 allo-HSCT donors who received Neupogen (n = 345, 64%) or Nivestym (n = 196, 36%) for PBSC mobilization. The median age was 47 years (range 17-76). The median donor weight was 86 kg (95% confidence interval [CI]: 87-91). Donors receiving Neupogen had similar pre-G-CSF white blood cell count, CD34+ percentages, and circulating CD34+ count compared with donors receiving Nivestym. The Neupogen group had similar median PBSC product total neutrophil count, CD34+ percentage, absolute CD34+ count, and infused CD34+ dose compared with the Nivestym group. For donors aged 35 years or younger, the median CD34+ dose was higher in donors who received Neupogen compared with Nivestym (6.9 vs. 6.3 million cells/kg, p = .044). CONCLUSIONS: Nivestym demonstrated similar efficacy for PBSC mobilization compared with Neupogen among allo-HSCT donors. In donors aged 35 years or younger, a slightly lower PBSC product CD34+ count was noted with Nivestym compared with Neupogen.
Asunto(s)
Biosimilares Farmacéuticos , Filgrastim , Movilización de Célula Madre Hematopoyética , Trasplante de Células Madre Hematopoyéticas , Células Madre de Sangre Periférica , Humanos , Filgrastim/uso terapéutico , Filgrastim/administración & dosificación , Filgrastim/farmacología , Adulto , Persona de Mediana Edad , Movilización de Célula Madre Hematopoyética/métodos , Masculino , Femenino , Estudios Retrospectivos , Anciano , Adolescente , Adulto Joven , Células Madre de Sangre Periférica/efectos de los fármacos , Trasplante Homólogo , Trasplante de Células Madre de Sangre PeriféricaRESUMEN
BACKGROUND: Germ cell tumors represent, among solid cancers, a potentially curable disease even if up to 20% to 30% of patients (pts) relapse after first-line treatment especially considering intermediate and poor prognosis groups. In this scenario, patients are candidates for high-dose chemotherapy and autologous stem-cells transplantation as second-line treatment even though stem-cells mobilization potential can be affected by several cycles and regimens of chemotherapy. To date, plerixafor is authorized in poor mobilizer adult pts diagnosed with lymphoma or multiple myeloma and in pediatric solid tumors or lymphoma. Therefore, the use of plerixafor in adult pts with relapsing/refractory GCT is still off label. MATERIALS AND METHODS: In our study, we describe mobilization and collection of peripheral blood stem cells for 10 pts with germ cell tumors. Six patients underwent plerixafor administration since classified as poor mobilizers based on WBC count (>5.000/µL) and CD34+ cell count (<15/µL) the day before apheresis procedure. RESULTS: On the first day of apheresis, plerixafor administration in poor mobilizers made possible a remarkable boost of CD34+ cells in such a way to overlap that of good mobilizers' (32/µL vs 35/µL, respectively, P > .05). CONCLUSION: Therefore, in our experience, plerixafor made a good fraction of poor mobilizer patients eligible for mobilization and collection and able to undergo the predicted autologous stem-cells transplantation; thus, the lack of access to the use of plerixafor in this setting of patients risks jeopardizing an effective treatment, especially in case of poor prognosis.
Asunto(s)
Bencilaminas/uso terapéutico , Ciclamas/uso terapéutico , Recurrencia Local de Neoplasia/terapia , Neoplasias de Células Germinales y Embrionarias/terapia , Trasplante de Células Madre de Sangre Periférica , Adolescente , Adulto , Bencilaminas/farmacología , Eliminación de Componentes Sanguíneos , Ciclamas/farmacología , Femenino , Movilización de Célula Madre Hematopoyética , Humanos , Masculino , Persona de Mediana Edad , Células Madre de Sangre Periférica/efectos de los fármacos , Estudios Retrospectivos , Trasplante Autólogo , Adulto JovenRESUMEN
PURPOSE: The purpose of this study was to evaluate mobilization outcomes with biosimilar pegfilgrastim versus filgrastim in association with chemotherapy as a mobilization strategy for lymphoma patients. METHODS: In the current study we included 32 lymphoma patients that received mobilization therapy and PBSC harvesting at the Bone Marrow Transplantation Department of Fundeni Clinical Institute, Bucharest, Romania between January and December 2019. RESULTS: Pegfilgrastim had beneficial effect when compared to filgrastim in reducing grade IV neutropenia both in the univariate and multivariate logistic models. Additionally, similar efficacy, as mobilization rate, after both filgrastim and pegfilgrastim was observed and no differences were noted between the two groups considering the need for platelet or red blood cell support. CONCLUSION: The use of biosimilar pegfilgrastim is a viable alternative to filgrastim in PBSC mobilization for lymphoma patients.
Asunto(s)
Filgrastim/farmacología , Filgrastim/uso terapéutico , Fármacos Hematológicos/farmacología , Fármacos Hematológicos/uso terapéutico , Movilización de Célula Madre Hematopoyética , Linfoma/tratamiento farmacológico , Células Madre de Sangre Periférica/efectos de los fármacos , Polietilenglicoles/farmacología , Polietilenglicoles/uso terapéutico , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios ProspectivosRESUMEN
BACKGROUND: Pleraxifor for peripheral blood stem cell (PBSC) mobilization in children with malignancies is often given following failure of standard mobilization (SM) rather than as a primary mobilizing agent. STUDY DESIGN AND METHODS: In this retrospective multicenter study, we report the safety of plerixafor-based PBSC mobilization in children with malignancies and compare outcomes between patients who received plerixafor upfront with SM (Group A) with those who received plerixafor following failure of SM (Group B). In the latter pleraxifor was given either following a low peripheral blood (PB) CD34 (<20 cells/cu.mm) (Group B1) or as a second collection process due to an unsuccessful yield (CD34 + < 2 × 106 /kg) (Group B2) following failed SM and first apheresis attempts. RESULTS: The study cohort (n = 47) with a median age of 8 (range 0.6-21) year, comprised 19 (40%) Group A and 28 (60%) Group B patients (B1 = 12 and B2 = 16). Pleraxifor mobilization was successful in 87.2% of patients, similar between Groups A and B (84.2% vs 89.2%) and resulted in a median 4-fold increase in PB CD34. Median number of apheresis attempts was 2 in Groups A and B1 but 4 in Group B2. In Group B2, median total CD34+ yield post-plerixafor was 9-fold higher than after SM (P = .0013). Mild to moderate transient adverse events affected 8.5% of patients. Among patients who proceeded to autologous transplant (n = 39), all but one engrafted. CONCLUSION: Plerixafor-based PBSC collection was safe and effective in our cohort and supports consideration as a primary mobilizing agent in children with malignancies.
Asunto(s)
Bencilaminas/uso terapéutico , Ciclamas/uso terapéutico , Movilización de Célula Madre Hematopoyética/métodos , Neoplasias/tratamiento farmacológico , Neoplasias/terapia , Células Madre de Sangre Periférica/efectos de los fármacos , Adolescente , Antígenos CD34/sangre , Eliminación de Componentes Sanguíneos , Niño , Preescolar , Estudios de Cohortes , Femenino , Humanos , Lactante , Linfoma/tratamiento farmacológico , Linfoma/terapia , Masculino , Meduloblastoma/tratamiento farmacológico , Meduloblastoma/radioterapia , Meduloblastoma/terapia , Neuroblastoma/tratamiento farmacológico , Neuroblastoma/radioterapia , Neuroblastoma/terapia , Células Madre de Sangre Periférica/metabolismo , Estudios Retrospectivos , Sarcoma/tratamiento farmacológico , Sarcoma/terapia , Adulto JovenRESUMEN
Graft-versus host disease (GVHD) remains one of the main causes of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (ASCT). Prophylactic T cell depletion via antithymocyte globulin (ATG) during ASCT conditioning is one of the standards of care for GVHD prophylaxis, although the optimal dosing strategy is still unclear. Recent studies have reported that absolute lymphocyte count at the time of ATG administration could predict survivals in ASCT from unrelated donors. Here this issue was examined in 116 patients receiving peripheral blood stem cells (PBSC) ASCT with purine analog/busulfan-based conditioning regimens between 2009 and 2019 in our department. The impact of lymphopenia at the time of ATG administration was evaluated in terms of overall survival, disease-free survival and GVHD-free/relapse-free survival. After a median follow-up of 4 years, no adverse effect of a profound lymphopenia was observed on patients' outcome. Notably, a reduced dose of ATG in patients with profound lymphopenia did not translate into better survivals. This study indicates that ATG can be administered whatever the recipient's lymphocyte counts in patients receiving a PBSC purine analog/busulfan-based conditioning regimen ASCT.
Asunto(s)
Suero Antilinfocítico/uso terapéutico , Linfopenia/tratamiento farmacológico , Trasplante de Células Madre de Sangre Periférica/métodos , Adulto , Anciano , Aloinjertos , Suero Antilinfocítico/administración & dosificación , Suero Antilinfocítico/metabolismo , Busulfano/administración & dosificación , Supervivencia sin Enfermedad , Femenino , Enfermedad Injerto contra Huésped/etiología , Trasplante de Células Madre Hematopoyéticas/mortalidad , Humanos , Depleción Linfocítica , Linfopenia/etiología , Masculino , Persona de Mediana Edad , Células Madre de Sangre Periférica/efectos de los fármacos , Nucleósidos de Purina/administración & dosificación , Estudios Retrospectivos , Acondicionamiento Pretrasplante , Trasplante HomólogoRESUMEN
A higher incidence of graft-versus-host disease (GVHD) has been observed after haploidentical hematopoietic stem cell transplantation (h-HSCT) with posttransplant cyclophosphamide (PTCY) using peripheral blood stem cells (PBSC) as a source of graft. Moreover, combining PTCY with antithymocyte globulin (ATG) may help to reduce GVHD incidence. In this study, early immune reconstitution, especially of T and NK cell compartments, was compared after both types of transplant (PTCY versus PTCY + ATG) investigate their influence on patient outcomes. This retrospective study included 58 adults who received a reduced intensity conditioning to PBSC h-HSCT with cyclosporine and mycophenolate mofetyl + PTCY (n = 32) or PTCY + ATG (n = 26) as GVHD prophylaxis. Both groups shared similar characteristics except for the median number of CD3+ T cells infused, significantly higher for PTCY + ATG patients. Blood samples from all patients were collected three times a week from day 0 until day 30 then at day 60 and day 90/100 to evaluate T and NK cells reconstitution by flow cytometry. The results show that PTCY + ATG versus PTCY alone significantly limits the occurrence of acute grade 2-4 GVHD after reduced intensity conditioning PBSC h-HSCT, perhaps because of the combined effect of T and NK cell reconstitution. Indeed, although a slower T cell reconstitution with PTCY + ATG may limit GVHD occurrence, the quicker reconstitution of some NK cell subtypes may help with avoiding relapse. Larger prospective studies are needed to better determine which NK cell subsets may influence the incidence of relapse after h-HSCT and optimize donor selection.
Asunto(s)
Suero Antilinfocítico/uso terapéutico , Ciclofosfamida/uso terapéutico , Enfermedad Injerto contra Huésped/tratamiento farmacológico , Inmunosupresores/uso terapéutico , Células Asesinas Naturales/efectos de los fármacos , Células Madre de Sangre Periférica/efectos de los fármacos , Linfocitos T/efectos de los fármacos , Adulto , Anciano , Complejo CD3/metabolismo , Femenino , Enfermedad Injerto contra Huésped/metabolismo , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Humanos , Células Asesinas Naturales/metabolismo , Masculino , Persona de Mediana Edad , Células Madre de Sangre Periférica/metabolismo , Estudios Retrospectivos , Trasplante de Células Madre/efectos adversos , Linfocitos T/metabolismo , Acondicionamiento Pretrasplante/efectos adversos , Trasplante Haploidéntico/efectos adversos , Trasplante Homólogo/efectos adversos , Adulto JovenRESUMEN
Cultured human mast cells are a useful tool for research into innate immune responses as well as allergic mechanisms. Mast cells cultured from peripheral blood can provide information on immune mechanisms of known, selected individuals. With the method presented here, eight million mast cells can be cultured from ca. one million stem cells purified from one unit (450 mL) of human peripheral blood. Culture with IgE and IL4 optimizes an allergic phenotype of the mast cells.
Asunto(s)
Hipersensibilidad/inmunología , Mastocitos/citología , Células Madre de Sangre Periférica/citología , Fenotipo , Cultivo Primario de Células/métodos , Antígeno AC133/genética , Antígeno AC133/metabolismo , Capa Leucocitaria de la Sangre/citología , Células Cultivadas , Medios de Cultivo/química , Humanos , Hipersensibilidad/sangre , Inmunidad Innata , Inmunoglobulina E/inmunología , Inmunoglobulina E/farmacología , Interleucina-4/inmunología , Interleucina-4/farmacología , Mastocitos/efectos de los fármacos , Mastocitos/inmunología , Células Madre de Sangre Periférica/efectos de los fármacos , Células Madre de Sangre Periférica/inmunologíaRESUMEN
Haplo-identical transplant is being increasingly used in patients who do not have a readily available matched related or unrelated donor. Post-transplant cyclophosphamide's use due to its simplicity and documented efficacy has made this approach readily employable across diverse transplant centres across the globe. The outcomes of regimens used for conditioning in recipients of bone marrow are at times in variance to that from more commonly employed G-CSF mobilised peripheral stem cell (PBSC). This review highlights various conditioning regimens used in PBSC recipients, with emphasis on toxicities, practicalities and transplant related outcomes of relapse, non-relapse mortality and graft versus host disease.
Asunto(s)
Ciclofosfamida/administración & dosificación , Factor Estimulante de Colonias de Granulocitos/farmacología , Neoplasias Hematológicas/terapia , Trasplante de Células Madre de Sangre Periférica/métodos , Células Madre de Sangre Periférica/efectos de los fármacos , Acondicionamiento Pretrasplante/métodos , Haplotipos , Movilización de Célula Madre Hematopoyética/métodos , Inmunosupresores/administración & dosificación , Células Madre de Sangre Periférica/citología , Ensayos Clínicos Controlados Aleatorios como Asunto , Trasplante HomólogoRESUMEN
Congenital erythropoietic porphyria (CEP) is an inborn error of heme synthesis resulting from uroporphyrinogen III synthase (UROS) deficiency and the accumulation of nonphysiological porphyrin isomer I metabolites. Clinical features are heterogeneous among patients with CEP but usually combine skin photosensitivity and chronic hemolytic anemia, the severity of which is related to porphyrin overload. Therapeutic options include symptomatic strategies only and are unsatisfactory. One promising approach to treating CEP is to reduce the erythroid production of porphyrins through substrate reduction therapy by inhibiting 5-aminolevulinate synthase 2 (ALAS2), the first and rate-limiting enzyme in the heme biosynthetic pathway. We efficiently reduced porphyrin accumulation after RNA interference-mediated downregulation of ALAS2 in human erythroid cellular models of CEP disease. Taking advantage of the physiological iron-dependent posttranscriptional regulation of ALAS2, we evaluated whether iron chelation with deferiprone could decrease ALAS2 expression and subsequent porphyrin production in vitro and in vivo in a CEP murine model. Treatment with deferiprone of UROS-deficient erythroid cell lines and peripheral blood CD34+-derived erythroid cultures from a patient with CEP inhibited iron-dependent protein ALAS2 and iron-responsive element-binding protein 2 expression and reduced porphyrin production. Furthermore, porphyrin accumulation progressively decreased in red blood cells and urine, and skin photosensitivity in CEP mice treated with deferiprone (1 or 3 mg/mL in drinking water) for 26 weeks was reversed. Hemolysis and iron overload improved upon iron chelation with full correction of anemia in CEP mice treated at the highest dose of deferiprone. Our findings highlight, in both mouse and human models, the therapeutic potential of iron restriction to modulate the phenotype in CEP.
Asunto(s)
Anemia Hemolítica/tratamiento farmacológico , Deferiprona/uso terapéutico , Quelantes del Hierro/uso terapéutico , Sobrecarga de Hierro/tratamiento farmacológico , Trastornos por Fotosensibilidad/tratamiento farmacológico , Porfiria Eritropoyética/tratamiento farmacológico , 5-Aminolevulinato Sintetasa/antagonistas & inhibidores , 5-Aminolevulinato Sintetasa/biosíntesis , 5-Aminolevulinato Sintetasa/genética , Adulto , Anemia Hemolítica/etiología , Animales , Sistemas CRISPR-Cas , Línea Celular , Línea Celular Tumoral , Modelos Animales de Enfermedad , Células Eritroides/efectos de los fármacos , Células Eritroides/metabolismo , Femenino , Técnicas de Sustitución del Gen , Humanos , Hierro/metabolismo , Sobrecarga de Hierro/etiología , Leucemia Eritroblástica Aguda/patología , Ratones , Células Madre de Sangre Periférica/efectos de los fármacos , Células Madre de Sangre Periférica/metabolismo , Trastornos por Fotosensibilidad/etiología , Porfiria Intermitente Aguda/metabolismo , Porfiria Eritropoyética/complicaciones , Porfirinas/biosíntesis , Interferencia de ARN , ARN Interferente Pequeño/farmacologíaRESUMEN
The aim of the study was to establish an in vitro fracture hematoma (FH) model that mimics the in vivo situation of the human fracture gap in order to assess drug efficacy and effectiveness for the treatment of fracture healing disorders. Human peripheral blood and mesenchymal stromal cells (MSCs) were coagulated to produce in vitro FH models, which were incubated in osteogenic medium under normoxia/hypoxia and analyzed for cell composition, gene expression and cytokine/chemokine secretion. To evaluate the model, we studied the impact of dexamethasone (impairing fracture healing) and deferoxamine (promoting fracture healing). Under hypoxic conditions, MSCs represented the predominant cell population, while the frequencies of leukocyte populations decreased. Marker gene expression of osteogenesis, angiogenesis, inflammation, migration and hypoxic adaptation increased significantly over time and compared to normoxia, while cytokine/chemokine secretion remained unchanged. Dexamethasone favored the frequency of immune cells compared to MSCs, suppressed osteogenic and pro-angiogenic gene expression, and enhanced the secretion of inflammatory cytokines. Conversely, deferoxamine favored the frequency of MSCs over that of immune cells and enhanced the expression of the osteogenic marker RUNX2 and markers of hypoxic adaptation. In summary, we demonstrate that hypoxia is an important factor for modeling the initial phase of fracture healing in vitro and that both fracture-healing disrupting and promoting substances can influence the in vitro model comparable to the in vivo situation. Therefore, we conclude that our model is able to mimic in part the human FH and could reduce the number of animal experiments in early preclinical studies.
Asunto(s)
Deferoxamina/farmacología , Dexametasona/farmacología , Fracturas Óseas/patología , Hematoma/patología , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre de Sangre Periférica/efectos de los fármacos , Alternativas a las Pruebas en Animales , Antiinflamatorios/farmacología , Humanos , Modelos Biológicos , Sideróforos/farmacología , Factores de TiempoRESUMEN
Gene rearrangements of MLL/KMT2A or RUNX1 are the major cause of therapy-related leukemia. Moreover, MLL rearrangements are the major cause of infant leukemia, and RUNX1 rearrangements are frequently detected in cord blood. These genes are sensitive to topoisomerase II inhibitors, and various genes have been identified as potential fusion partners. However, fetal exposure to these inhibitors is rare. Therefore, we postulated that even a proliferation signal itself might induce gene rearrangements in hematopoietic stem cells. To test this hypothesis, we detected gene rearrangements in etoposide-treated or non-treated CD34+ cells cultured with cytokines using inverse PCR. In the etoposide-treated cells, variable-sized rearrangement bands were detected in the RUNX1 and MLL genes at 3 hours of culture, which decreased after 7 days. However, more rearrangement bands were detected in the non-treated cells at 7 days of culture. Such gene rearrangements were also detected in peripheral blood stem cells mobilized by cytokines for transplantation. However, none of these rearranged genes encoded the leukemogenic oncogene, and the cells with rearrangements did not expand. These findings suggest that MLL and RUNX1 rearrangements, which occur with very low frequency in normal hematopoietic progenitor cells, may be induced under cytokine stimulation. Most of the cells with gene rearrangements are likely eliminated, except for leukemia-associated gene rearrangements, resulting in the low prevalence of leukemia development.
Asunto(s)
Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Citocinas/farmacología , Reordenamiento Génico/efectos de los fármacos , Células Madre Hematopoyéticas/efectos de los fármacos , N-Metiltransferasa de Histona-Lisina/genética , Proteína de la Leucemia Mieloide-Linfoide/genética , Anciano , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Etopósido/farmacología , Células Madre Hematopoyéticas/metabolismo , Humanos , Linfoma de Células B Grandes Difuso/patología , Persona de Mediana Edad , Células Madre de Sangre Periférica/efectos de los fármacos , Células Madre de Sangre Periférica/metabolismo , Inhibidores de Topoisomerasa II/farmacologíaRESUMEN
BACKGROUND: Donor factors have a variable correlation with cluster of differentiation (CD)34+ cell dose in allogeneic peripheral blood stem cell (PBSC) harvests. CD34+ cell dose affects the speed of hematopoietic recovery and percentage of donor chimerism in the recipient. METHODS: A total of 25 allogeneic PBSC transplants performed during a 3-year period were included. All donors underwent mobilization with filgrastim. Leukapheresis, flowcytometric CD34+ cell enumeration, and chimerism analysis were performed and correlated with recipient outcome. RESULTS: Besides age, all other donor parameters had a positive correlation with CD34+ cell count. Engraftment kinetics and chimerism had a positive correlation with the CD34+ yield of the PBSC product. Acute graft-vs-host disease (GVHD) was observed in patients with complete chimerism at day 30 after transplantation. CONCLUSION: Adequate CD34+ cell yield happens in healthy donors, independent of donor demographic patterns with G-CSF only. A diverse population of donors can thus be approached for Matched Unrelated Donor (MUD) transplants. An accurate quantitative analysis of early donor chimerism in the recipient (at day 30) is an excellent tool for post-transplant monitoring for acute GvHD.
Asunto(s)
Donantes de Sangre/estadística & datos numéricos , Enfermedad Injerto contra Huésped/epidemiología , Trasplante de Células Madre de Sangre Periférica/efectos adversos , Factores de Edad , Antígenos CD34/genética , Antígenos CD34/metabolismo , Filgrastim/farmacología , Factor Estimulante de Colonias de Granulocitos/metabolismo , Fármacos Hematológicos/farmacología , Humanos , Trasplante de Células Madre de Sangre Periférica/métodos , Trasplante de Células Madre de Sangre Periférica/estadística & datos numéricos , Células Madre de Sangre Periférica/efectos de los fármacos , Células Madre de Sangre Periférica/metabolismo , Trasplante HomólogoRESUMEN
Millions of women are exposed simultaneously to antiretroviral drugs (ARVs) and progestin-based hormonal contraceptives. Yet the reciprocal modulation by ARVs and progestins of their intracellular functions is relatively unexplored. We investigated the effects of tenofovir disoproxil fumarate (TDF) and dapivirine (DPV), alone and in the presence of select steroids and progestins, on cell viability, steroid-regulated immunomodulatory gene expression, activation of steroid receptors, and anti-HIV-1 activity in vitro Both TDF and DPV modulated the transcriptional efficacy of a glucocorticoid agonist via the glucocorticoid receptor (GR) in the U2OS cell line. In TZM-bl cells, DPV induced the expression of the proinflammatory interleukin 8 (IL-8) gene while TDF significantly increased medroxyprogesterone acetate (MPA)-induced expression of the anti-inflammatory glucocorticoid-induced leucine zipper (GILZ) gene. However, peripheral blood mononuclear cell (PBMC) and ectocervical explant tissue viability and gene expression results, along with TZM-bl HIV-1 infection data, are reassuring and suggest that TDF and DPV, in combination with dexamethasone (DEX) or MPA, do not reciprocally modulate key biological effects in primary cells and tissue. We show for the first time that TDF induces progestogen-independent activation of the progesterone receptor (PR) in a cell line. The ability of TDF and DPV to influence GR and PR activity suggests that their use may be associated with steroid receptor-mediated off-target effects. This, together with cell line and individual donor gene expression responses in the primary models, raises concerns that reciprocal modulation may cause side effects in a cell- and donor-specific manner in vivo.
Asunto(s)
Antirretrovirales/efectos adversos , Antirretrovirales/farmacología , Receptores de Esteroides/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Humanos , Inflamación/metabolismo , Células Madre de Sangre Periférica/efectos de los fármacos , Células Madre de Sangre Periférica/metabolismo , Progestinas/metabolismo , Pirimidinas/efectos adversos , Pirimidinas/farmacología , Receptores de Glucocorticoides/metabolismo , Receptores de Progesterona/metabolismo , Tenofovir/efectos adversos , Tenofovir/farmacología , Factores de Transcripción/metabolismoRESUMEN
Hydroxyurea (HU) is a nonalkylating antineoplastic agent used in the treatment of hematological malignancies. HU is a DNA replication stress inducer, and as such, it may induce a premature senescence-like cell phenotype; however, its repercussion on bystander cell proliferation has not been revealed so far. Our results indicate that HU strongly inhibited peripheral blood mesenchymal stromal cells (PBMSC) proliferation by cell cycle arrest in S phase, and that, consequently, PBMSC acquire senescence-related phenotypical changes. HU-treated PBMSC display increased senescence-associated ß-galactosidase levels and p16INK4 expression, as well as DNA damage response and genotoxic effects, evidenced by expression of γH2A.X and micronuclei. Moreover, HU-induced PBMSC senescence is mediated by increased reactive oxygen species (ROS) levels, as demonstrated by the inhibition of senescence markers in the presence of ROS scavenger N-acetylcysteine and NADPH oxidase inhibitor Apocynin. To determine the HU-induced bystander effect, we used the JAK2V617F-positive human erythroleukemia 92.1.7 (HEL) cells. Co-culture with HU-induced senescent PBMSC (HU-S-PBMSC) strongly inhibited bystander HEL cell proliferation, and this effect is mediated by both ROS and transforming growth factor (TGF)-ß expression. Besides induction of premature senescence, HU educates PBMSC toward an inhibitory phenotype of HEL cell proliferation. Finally, our study contributes to the understanding of the role of HU-induced PBMSC senescence as a potential adjuvant in hematological malignancy therapies.
Asunto(s)
Senescencia Celular/efectos de los fármacos , Hidroxiurea/farmacología , Janus Quinasa 2/genética , Leucemia Eritroblástica Aguda/tratamiento farmacológico , Factor de Crecimiento Transformador beta/genética , Efecto Espectador/genética , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Leucemia Eritroblástica Aguda/genética , Leucemia Eritroblástica Aguda/patología , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre de Sangre Periférica/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Interleukin-6 (IL-6) contributes to the development of immune-mediated complications after allogeneic stem cell transplantation. However, systemic IL-6 levels also increase during granulocyte colony-stimulating factor (G-CSF) mobilization of hematopoietic stem cells in healthy donors, but it is not known whether this mobilization alters systemic levels of other IL-6 family cytokines/receptors and whether such effects differ between donors. We examined how G-CSF administration influenced C-reactive protein (CRP) levels (85 donors) and serum levels of IL-6 family cytokines/receptors (20 donors). G-CSF increased CRP levels especially in elderly donors with high pretherapy levels, but these preharvesting levels did not influence clinical outcomes (nonrelapse mortality, graft versus host disease). The increased IL-6 levels during G-CSF therapy normalized within 24 h after treatment. G-CSF administration did not alter serum levels of other IL-6-familly mediators. Oncostatin M, but not IL-6, showed a significant correlation with CRP levels during G-CSF therapy. Clustering analysis of mediator levels during G-CSF administration identified two donor subsets mainly characterized by high oncostatin M and IL-6 levels, respectively. Finally, G-CSF could increase IL-6 release by in vitro cultured monocytes, fibroblasts, and mesenchymal stem cells. In summary, G-CSF seems to induce an acute phase reaction with increased systemic IL-6 levels in healthy stem cell donors.
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Donantes de Sangre , Fibroblastos/inmunología , Factor Estimulante de Colonias de Granulocitos/farmacología , Mediadores de Inflamación/metabolismo , Inflamación/inmunología , Monocitos/inmunología , Células Madre de Sangre Periférica/inmunología , Adolescente , Adulto , Anciano , Células Cultivadas , Citocinas/metabolismo , Femenino , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Movilización de Célula Madre Hematopoyética , Humanos , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Persona de Mediana Edad , Monocitos/efectos de los fármacos , Monocitos/metabolismo , Células Madre de Sangre Periférica/efectos de los fármacos , Células Madre de Sangre Periférica/metabolismo , Adulto JovenRESUMEN
It has been estimated that up to a third of global malaria deaths may be attributable to malarial anaemia, with children and pregnant women being those most severely affected. An inefficient erythropoietic response to the destruction of both infected and uninfected erythrocytes in infections with Plasmodium spp. contributes significantly to the development and persistence of such anaemia. The underlying mechanisms, which could involve both direct inhibition of erythropoiesis by parasite-derived factors and indirect inhibition as a result of modulation of the immune response, remain poorly understood. We found parasite-derived factors in conditioned medium (CM) of blood-stage Plasmodium falciparum and crude isolates of parasite haemozoin directly to inhibit erythropoiesis in an ex vivo model based on peripheral blood haematopoietic stem cells. Erythropoiesis-inhibiting activity was detected in a fraction of CM that was sensitive to heat inactivation and protease digestion. Erythropoiesis was also inhibited by crude parasite haemozoin but not by detergent-treated, heat-inactivated or protease-digested haemozoin. These results suggest that the erythropoiesis-inhibiting activity in both cases is mediated by proteins or protein-containing biomolecules and may offer new leads to the treatment of malarial anaemia.
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Medios de Cultivo Condicionados/farmacología , Eritrocitos/efectos de los fármacos , Eritropoyesis/efectos de los fármacos , Hemoproteínas/farmacología , Plasmodium falciparum/química , Células Cultivadas , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Péptido Hidrolasas/metabolismo , Células Madre de Sangre Periférica/efectos de los fármacosRESUMEN
In short-term animal models of ischemia, erythropoietin (EPO) signaling through the heterodimeric EPO receptor (EPOR)/ß-common receptor (ßCR) is believed to elicit tissue protective effects. However, large, randomized, controlled trials demonstrate that targeting a higher hemoglobin level by administering higher doses of EPO, which are more likely to activate the heterodimeric EPOR/ßCR, is associated with an increase in adverse cardiovascular events. Thus, inhibition of long-term activation of the ßCR may have therapeutic implications. This study aimed to design and evaluate the efficacy of novel computationally designed ßCR inhibitory peptides (ßIP). These novel ßIPs were designed based on a truncated portion of Helix-A from EPO, specifically residues 11-26 (VLERYLLEAKEAEKIT). Seven novel peptides (P1 to P7) were designed. Peptide 7 (P7), VLERYLHEAKHAEKIT, demonstrated the most robust inhibitory activity. We also report here the ability of P7 to inhibit ßCR-induced nitric oxide (NO) production and angiogenesis in human umbilical vein endothelial cells (HUVECs). Specifically, we found that P7 ßIP completely abolished EPO-induced NO production. The inhibitory effect could be overcome with super physiological doses of EPO, suggesting a competitive inhibition. ßCR-induced angiogenesis in HUVEC's was also abolished with treatment of P7 ßIP, but P7 ßIP did not inhibit vascular endothelial growth factor (VEGF)-induced angiogenesis. In addition, we demonstrate that the novel P7 ßIP does not inhibit EPO-induced erythropoiesis with use of peripheral blood mononuclear cells (PBMCs). These results, for the first time, describe a novel, potent ßCR peptide inhibitor that inhibit the actions of the ßCR without affecting erythropoiesis.
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Subunidad beta Común de los Receptores de Citocinas/metabolismo , Eritropoyetina/farmacología , Células Madre de Sangre Periférica/metabolismo , Transducción de Señal/fisiología , Secuencia de Aminoácidos , Células Cultivadas , Biología Computacional , Células Endoteliales de la Vena Umbilical Humana , Humanos , Datos de Secuencia Molecular , Óxido Nítrico/metabolismo , Células Madre de Sangre Periférica/efectos de los fármacos , Estructura Secundaria de Proteína , Transducción de Señal/genéticaRESUMEN
Bortezomib-based induction is often used in transplant-eligible patients with myeloma. The optimal peripheral blood stem cell (PBSC) mobilisation strategy in this context is unclear. We reviewed the efficacy of G-CSF alone (G-alone) vs. G-CSF and cyclophosphamide (G-cyclo: standard dose: 1.5-2 g/m2; high dose: 3-4 g/m2) PBSC mobilisation strategies in 288 patients who only received bortezomib, cyclophosphamide and dexamethasone (VCD) induction prior to autograft across six apheresis centres from November 2012 to June 2017. 'Uncomplicated successful mobilisation' was defined as achieving a PBSC yield of ≥4 × 106/kg within two aphereses, without plerixafor or mobilisation-associated toxicity (predominantly febrile neutropenia, FN). Success rates were 84% in G-cyclo standard dose (6% FN), 64% in G-cyclo high dose (18% FN) and 69% in G-alone (plerixafor successfully salvaged 8/9 patients). Median total stem cell yield was significantly higher with G-cyclo, but not different between the two cyclophosphamide doses. Age greater than the median of 61 years was associated with higher failure rates (22 vs. 11%, p = 0.01) and lower PBSC yield, especially in the G-alone group. Prior radiotherapy exposure did not impact on collection success. Our observations suggest that both G-cyclo standard dose and G-alone are reasonable mobilisation strategies. The former may be preferred if salvage plerixafor is unavailable.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos/administración & dosificación , Movilización de Célula Madre Hematopoyética/métodos , Mieloma Múltiple/terapia , Células Madre de Sangre Periférica/efectos de los fármacos , Adulto , Factores de Edad , Anciano , Bortezomib/uso terapéutico , Ciclofosfamida/administración & dosificación , Dexametasona/uso terapéutico , Movilización de Célula Madre Hematopoyética/normas , Humanos , Masculino , Persona de Mediana Edad , Células Madre de Sangre Periférica/citología , Inducción de Remisión/métodosRESUMEN
BACKGROUND: Peripheral blood progenitor cell (PBPC) mobilization with chemotherapy in addition to Granulocyte-Colony Stimulating Factor (G-CSF) improves cell collection compared to G-CSF alone; however, it is associated with increased risk of neutropenic fever (NF). METHODS: We analyzed risk factors for post-priming NF and NF association with autologous stem cell transplant outcomes. Between 1998 and 2008, 593 adult patients with lymphoma underwent PBPC mobilization with etoposide and G-CSF. RESULTS: Median age was 51 years (range 18-77) and 372 (63%) were male. Diagnoses were 457 (77%) non-Hodgkin lymphoma and 136 (23%) Hodgkin lymphoma. Of 554 (93%) transplanted patients, majority were in complete or partial remission at time of transplant (88%). Overall, 141 (24%) patients were hospitalized for NF. Nine patients (6%) had bacteremia, 4 (3%) had pneumonia, 2 (<1%) had herpes simplex viral infections, and the remaining 126 (90%) had no identified infection source. NF patients had lower likelihood of proceeding to transplant (86% vs. 96%, P < .001), lower CD34+ cell dose collection (median 7.23 × 106 CD34+ cells/kg vs. 8.98 × 106 CD34+ cells/kg, P = .002), and were more likely to require > 4 days of apheresis (48% vs. 37%, P < .001). NF was associated with a higher 30-day readmission rate following transplant hospitalization (17% vs. 9%, P = .012). CONCLUSION: NF during etoposide priming is associated with lower likelihood of proceeding to transplant, lower CD34+ cell dose collection, more apheresis days required for collection and a higher 30-day readmission rate following transplant discharge.