Search

Chemotherapy-induced mucositis, characterized by diarrhoea and villous atrophy, is a severe side effect contributing to reduced quality of life and premature death in cancer patients treated with cytostatics. Despite its high incidence, there is no effective supportive therapy available. The main objective of this study was to determine if the anti-inflammatory drugs anakinra and/or dexamethasone—which have different mechanisms-of-action—might be used to effectively treat idarubicin-induced mucositis in rats. Mucositis was induced through a single injection with 2 mg/kg idarubicin (with saline as control), followed by daily treatments of anakinra (100 mg/kg/day), dexamethasone (10 mg/kg/day) or both for 3 days. After 72 h, jejunal tissue was collected for morphological, apoptotic and proliferative analyses, and colonic faecal water content and body weight change were determined. The diarrhoea that was induced by idarubicin (from 63.5% to 78.6% water content in faeces) was completely reversed by anakinra alone, and the jejunal villus height reduction by 36% was prevented by a combination of anakinra and dexamethasone. Dexamethasone reduced apoptosis in the jejunal crypts, both alone and in combination with anakinra. These positive effects encouraged further investigations into the use of anakinra and dexamethasone as supportive therapies for chemotherapy-induced intestinal mucositis and diarrhoea., Authors and title in the list of papers of Fredrik Kullenberg's thesis:Kullenberg, F., Peters, K., Dahlgren, D., Heindryckx, F., Sjöblom, M., & Lennernäs, H.Anakinra and dexamethasone treatment of idarubicin-induced mucositis and diarrhea in rats

Chemotherapy-induced mucositis, characterized by diarrhoea and villous atrophy, is a severe side effect contributing to reduced quality of life and premature death in cancer patients treated with cytostatics. Despite its high incidence, there is no effective supportive therapy available. The main objective of this study was to determine if the anti-inflammatory drugs anakinra and/or dexamethasone—which have different mechanisms-of-action—might be used to effectively treat idarubicin-induced mucositis in rats. Mucositis was induced through a single injection with 2 mg/kg idarubicin (with saline as control), followed by daily treatments of anakinra (100 mg/kg/day), dexamethasone (10 mg/kg/day) or both for 3 days. After 72 h, jejunal tissue was collected for morphological, apoptotic and proliferative analyses, and colonic faecal water content and body weight change were determined. The diarrhoea that was induced by idarubicin (from 63.5% to 78.6% water content in faeces) was completely reversed by anakinra alone, and the jejunal villus height reduction by 36% was prevented by a combination of anakinra and dexamethasone. Dexamethasone reduced apoptosis in the jejunal crypts, both alone and in combination with anakinra. These positive effects encouraged further investigations into the use of anakinra and dexamethasone as supportive therapies for chemotherapy-induced intestinal mucositis and diarrhoea., Authors and title in the list of papers of Fredrik Kullenberg's thesis:Kullenberg, F., Peters, K., Dahlgren, D., Heindryckx, F., Sjöblom, M., & Lennernäs, H.Anakinra and dexamethasone treatment of idarubicin-induced mucositis and diarrhea in rats

Chemotherapy-induced mucositis, characterized by diarrhoea and villous atrophy, is a severe side effect contributing to reduced quality of life and premature death in cancer patients treated with cytostatics. Despite its high incidence, there is no effective supportive therapy available. The main objective of this study was to determine if the anti-inflammatory drugs anakinra and/or dexamethasone—which have different mechanisms-of-action—might be used to effectively treat idarubicin-induced mucositis in rats. Mucositis was induced through a single injection with 2 mg/kg idarubicin (with saline as control), followed by daily treatments of anakinra (100 mg/kg/day), dexamethasone (10 mg/kg/day) or both for 3 days. After 72 h, jejunal tissue was collected for morphological, apoptotic and proliferative analyses, and colonic faecal water content and body weight change were determined. The diarrhoea that was induced by idarubicin (from 63.5% to 78.6% water content in faeces) was completely reversed by anakinra alone, and the jejunal villus height reduction by 36% was prevented by a combination of anakinra and dexamethasone. Dexamethasone reduced apoptosis in the jejunal crypts, both alone and in combination with anakinra. These positive effects encouraged further investigations into the use of anakinra and dexamethasone as supportive therapies for chemotherapy-induced intestinal mucositis and diarrhoea., Authors and title in the list of papers of Fredrik Kullenberg's thesis:Kullenberg, F., Peters, K., Dahlgren, D., Heindryckx, F., Sjöblom, M., & Lennernäs, H.Anakinra and dexamethasone treatment of idarubicin-induced mucositis and diarrhea in rats

Chemotherapy-induced mucositis, characterized by diarrhoea and villous atrophy, is a severe side effect contributing to reduced quality of life and premature death in cancer patients treated with cytostatics. Despite its high incidence, there is no effective supportive therapy available. The main objective of this study was to determine if the anti-inflammatory drugs anakinra and/or dexamethasone—which have different mechanisms-of-action—might be used to effectively treat idarubicin-induced mucositis in rats. Mucositis was induced through a single injection with 2 mg/kg idarubicin (with saline as control), followed by daily treatments of anakinra (100 mg/kg/day), dexamethasone (10 mg/kg/day) or both for 3 days. After 72 h, jejunal tissue was collected for morphological, apoptotic and proliferative analyses, and colonic faecal water content and body weight change were determined. The diarrhoea that was induced by idarubicin (from 63.5% to 78.6% water content in faeces) was completely reversed by anakinra alone, and the jejunal villus height reduction by 36% was prevented by a combination of anakinra and dexamethasone. Dexamethasone reduced apoptosis in the jejunal crypts, both alone and in combination with anakinra. These positive effects encouraged further investigations into the use of anakinra and dexamethasone as supportive therapies for chemotherapy-induced intestinal mucositis and diarrhoea., Authors and title in the list of papers of Fredrik Kullenberg's thesis:Kullenberg, F., Peters, K., Dahlgren, D., Heindryckx, F., Sjöblom, M., & Lennernäs, H.Anakinra and dexamethasone treatment of idarubicin-induced mucositis and diarrhea in rats

Chemotherapy-induced mucositis, characterized by diarrhoea and villous atrophy, is a severe side effect contributing to reduced quality of life and premature death in cancer patients treated with cytostatics. Despite its high incidence, there is no effective supportive therapy available. The main objective of this study was to determine if the anti-inflammatory drugs anakinra and/or dexamethasone—which have different mechanisms-of-action—might be used to effectively treat idarubicin-induced mucositis in rats. Mucositis was induced through a single injection with 2 mg/kg idarubicin (with saline as control), followed by daily treatments of anakinra (100 mg/kg/day), dexamethasone (10 mg/kg/day) or both for 3 days. After 72 h, jejunal tissue was collected for morphological, apoptotic and proliferative analyses, and colonic faecal water content and body weight change were determined. The diarrhoea that was induced by idarubicin (from 63.5% to 78.6% water content in faeces) was completely reversed by anakinra alone, and the jejunal villus height reduction by 36% was prevented by a combination of anakinra and dexamethasone. Dexamethasone reduced apoptosis in the jejunal crypts, both alone and in combination with anakinra. These positive effects encouraged further investigations into the use of anakinra and dexamethasone as supportive therapies for chemotherapy-induced intestinal mucositis and diarrhoea., Authors and title in the list of papers of Fredrik Kullenberg's thesis:Kullenberg, F., Peters, K., Dahlgren, D., Heindryckx, F., Sjöblom, M., & Lennernäs, H.Anakinra and dexamethasone treatment of idarubicin-induced mucositis and diarrhea in rats

A highly sensitive electrochemical DNA sensor for detection of the chemotherapeutic drug idarubicin mediated by Methylene blue (MB) has been developed. DNA from fish sperm has been immobilized at the electropolymerized layers of Azure B. The incorporation of MB into the DNA layers substantially increased the sensor sensitivity. The concentration range for idarubicin determination by cyclic voltammetry was from 1 fM to 0.1 nM, with a limit of detection (LOD) of 0.3 fM. Electrochemical impedance spectroscopy (EIS) in the presence of a redox probe ([Fe(CN)6 ]3−/4− ) allowed for the widening of a linear range of idarubicin detection from 1 fM to 100 nM, retaining LOD 0.3 fM. The DNA sensor has been tested in various real and artificial biological fluids with good recovery ranging between 90–110%. The sensor has been successfully used for impedimetric idarubicin detection in medical preparation Zavedos® . The developed DNA biosensor could be useful for the control of the level of idarubicin during cancer therapy as well as for pharmacokinetics studies. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

Anthracyclines belong to a class of cytostatic compounds that are commonly used to treat various types of cancers, including lymphoma, breast cancer and primary liver cancer. The aim of this thesis was to study how two anthracyclines, doxorubicin and idarubicin, induced cytotoxicity in tumor cells, as well as their off-target effect on the gastrointestinal system. In Paper I, we exposed four different cancer cell lines to various concentrations of doxorubicin in two different formulations, and measured the cell viability and how much doxorubicin was taken up by the cells. We found that the cell lines differed in their uptake and sensitivity, and that the most resistant cell line had an intracellular exposure of doxorubicin that was about 100 times lower when compared to the more sensitive cell lines. To study the off-target gastrointestinal toxicity, we dosed rats with doxorubicin and studied how chemotherapy-induced mucositis developed during seven days in Paper II. The main effect parameter was intestinal villus atrophy, which was most severe after three days. This was preceded by an increased cell death and decreased proliferation in the crypts, which occurred within the first day after doxorubicin exposure. To study how different cytostatic drugs affected chemotherapy-induced mucositis, and to what extent they caused diarrhea, rats were dosed with one of six different chemotherapies, including doxorubicin and idarubicin, in Paper III. All selected chemotherapies caused similar villus atrophy three days after dosing, but only a third, including idarubicin, caused clear diarrhea. In paper IV the objective was to treat or prevent idarubicin-induced mucositis and diarrhea, by using the anti-inflammatory drugs anakinra and/or dexamethasone. Anakinra alone stopped the diarrhea, while the combination of anakinra and dexamethasone prevented villus atrophy. These positive effects encourage further investigations into the use of anakinra and dexamethasone as supportive th

Anthracyclines belong to a class of cytostatic compounds that are commonly used to treat various types of cancers, including lymphoma, breast cancer and primary liver cancer. The aim of this thesis was to study how two anthracyclines, doxorubicin and idarubicin, induced cytotoxicity in tumor cells, as well as their off-target effect on the gastrointestinal system. In Paper I, we exposed four different cancer cell lines to various concentrations of doxorubicin in two different formulations, and measured the cell viability and how much doxorubicin was taken up by the cells. We found that the cell lines differed in their uptake and sensitivity, and that the most resistant cell line had an intracellular exposure of doxorubicin that was about 100 times lower when compared to the more sensitive cell lines. To study the off-target gastrointestinal toxicity, we dosed rats with doxorubicin and studied how chemotherapy-induced mucositis developed during seven days in Paper II. The main effect parameter was intestinal villus atrophy, which was most severe after three days. This was preceded by an increased cell death and decreased proliferation in the crypts, which occurred within the first day after doxorubicin exposure. To study how different cytostatic drugs affected chemotherapy-induced mucositis, and to what extent they caused diarrhea, rats were dosed with one of six different chemotherapies, including doxorubicin and idarubicin, in Paper III. All selected chemotherapies caused similar villus atrophy three days after dosing, but only a third, including idarubicin, caused clear diarrhea. In paper IV the objective was to treat or prevent idarubicin-induced mucositis and diarrhea, by using the anti-inflammatory drugs anakinra and/or dexamethasone. Anakinra alone stopped the diarrhea, while the combination of anakinra and dexamethasone prevented villus atrophy. These positive effects encourage further investigations into the use of anakinra and dexamethasone as supportive th

Anthracyclines belong to a class of cytostatic compounds that are commonly used to treat various types of cancers, including lymphoma, breast cancer and primary liver cancer. The aim of this thesis was to study how two anthracyclines, doxorubicin and idarubicin, induced cytotoxicity in tumor cells, as well as their off-target effect on the gastrointestinal system. In Paper I, we exposed four different cancer cell lines to various concentrations of doxorubicin in two different formulations, and measured the cell viability and how much doxorubicin was taken up by the cells. We found that the cell lines differed in their uptake and sensitivity, and that the most resistant cell line had an intracellular exposure of doxorubicin that was about 100 times lower when compared to the more sensitive cell lines. To study the off-target gastrointestinal toxicity, we dosed rats with doxorubicin and studied how chemotherapy-induced mucositis developed during seven days in Paper II. The main effect parameter was intestinal villus atrophy, which was most severe after three days. This was preceded by an increased cell death and decreased proliferation in the crypts, which occurred within the first day after doxorubicin exposure. To study how different cytostatic drugs affected chemotherapy-induced mucositis, and to what extent they caused diarrhea, rats were dosed with one of six different chemotherapies, including doxorubicin and idarubicin, in Paper III. All selected chemotherapies caused similar villus atrophy three days after dosing, but only a third, including idarubicin, caused clear diarrhea. In paper IV the objective was to treat or prevent idarubicin-induced mucositis and diarrhea, by using the anti-inflammatory drugs anakinra and/or dexamethasone. Anakinra alone stopped the diarrhea, while the combination of anakinra and dexamethasone prevented villus atrophy. These positive effects encourage further investigations into the use of anakinra and dexamethasone as supportive th

Introduction. Acute promyelocytic leukemia (APL) is a rare subtype of acute myeloid leukemia (AML), characterized by a particularly aggressive clinical behavior, that in the absence of treatment is usually fatal. The objective of this work was to determine the clinical and cytogenetic characteristics of a cohort of patients with APL, in order to evaluate their relationship with the outcome and prognosis of these patients. Methodology. An observational, descriptive, retrospective study of patients older than 15 years with a diagnosis of APL treated at the Hospital Universitario San Vicente Fundación, between 2012 and 2020, was carried out. Results. A total of 32 patients were included. The mean age at diagnosis was 37 years, 84.4% of the patients had the t(15;17) in the karyotype, and 93.75% had positive FISH. 12.5% of cases had a complex karyotype. Mortality in the first 30 days was 15.6%, with bleeding being the most common cause of death. All patients who survived achieved complete remission (84.3%). In an average follow-up of 24 months, 14.8% of cases relapsed. In the bivariate analysis, a relationship was found between the male sex and having a complex karyotype (p<0.015). No relationship was found between complex karyotype and early mortality (p=0.358), nor between complex karyotype and relapse (p=0.052). Conclusions. We present the clinical and cytogenetic characteristics of a cohort of patients with APL in Colombia. Central nervous system bleeding was the main cause of early mortality, with all surviving patients achieving complete remission on induction therapy. Mortality, complete remission and relapse rates were similar to those reported by other Latin American series, but lower than studies from European countries. Contrary to what has been reported in other studies, no relationship was found between complex karyotype and early mortality or relapse., Introducción. La leucemia promielocítica aguda (LPA) es un subtipo poco frecuente de leucemia mieloide aguda (LMA), que se caracteriza por un comportamiento clínico particularmente agresivo, y en ausencia de tratamiento, su curso generalmente es fatal. El objetivo de este trabajo fue determinar las características clínicas y citogenéticas de una cohorte de pacientes con LPA, con la finalidad de evaluar su relación con las complicaciones, el pronóstico y el desenlace de estos pacientes. Metodología. Se realizó un estudio observacional, descriptivo, retrospectivo de los pacientes mayores de 15 años con diagnóstico de LPA, atendidos en el Hospital Universitario San Vicente Fundación, entre los años 2012 a 2020. Resultados. Un total de 32 pacientes fueron incluidos. La edad media del diagnóstico fue 37 años. El 84,4% de los pacientes tenía la traslocación (15;17) en el cariotipo, y el 93,75% tenían FISH positivo. El 12,5% de los casos tenían cariotipo complejo. La mortalidad en los primeros 30 días fue del 15,6%, siendo el sangrado la causa de muerte más frecuente. Todos los pacientes que sobrevivieron alcanzaron la remisión completa (84,3%). En un promedio de seguimiento de 24 meses, el 14,8% de los casos recayeron. En el análisis bivariado se encontró relación entre sexo masculino y tener cariotipo complejo (p=0,015). No se encontró relación entre cariotipo complejo y mortalidad temprana (p=0,358), tampoco entre cariotipocomplejo y recaída (p=0,052). Conclusiones. Se presentan las características clínicas y citogenéticas de una cohorte de pacientes con LPA en Colombia. El sangrado en el sistema nervioso central fue la principal causa de mortalidad temprana, todos los pacientes que sobrevivieron alcanzaron la remisión completa con la terapia de inducción. Las tasas de mortalidad, remisión completa y recaída fueron similares a las reportadas por otras series latinoamericanas, pero inferiores a estudios provenientes de países europeos. Contrario a lo reportado en otros e

Anthracyclines belong to a class of cytostatic compounds that are commonly used to treat various types of cancers, including lymphoma, breast cancer and primary liver cancer. The aim of this thesis was to study how two anthracyclines, doxorubicin and idarubicin, induced cytotoxicity in tumor cells, as well as their off-target effect on the gastrointestinal system. In Paper I, we exposed four different cancer cell lines to various concentrations of doxorubicin in two different formulations, and measured the cell viability and how much doxorubicin was taken up by the cells. We found that the cell lines differed in their uptake and sensitivity, and that the most resistant cell line had an intracellular exposure of doxorubicin that was about 100 times lower when compared to the more sensitive cell lines. To study the off-target gastrointestinal toxicity, we dosed rats with doxorubicin and studied how chemotherapy-induced mucositis developed during seven days in Paper II. The main effect parameter was intestinal villus atrophy, which was most severe after three days. This was preceded by an increased cell death and decreased proliferation in the crypts, which occurred within the first day after doxorubicin exposure. To study how different cytostatic drugs affected chemotherapy-induced mucositis, and to what extent they caused diarrhea, rats were dosed with one of six different chemotherapies, including doxorubicin and idarubicin, in Paper III. All selected chemotherapies caused similar villus atrophy three days after dosing, but only a third, including idarubicin, caused clear diarrhea. In paper IV the objective was to treat or prevent idarubicin-induced mucositis and diarrhea, by using the anti-inflammatory drugs anakinra and/or dexamethasone. Anakinra alone stopped the diarrhea, while the combination of anakinra and dexamethasone prevented villus atrophy. These positive effects encourage further investigations into the use of anakinra and dexamethasone as supportive th

Anthracyclines belong to a class of cytostatic compounds that are commonly used to treat various types of cancers, including lymphoma, breast cancer and primary liver cancer. The aim of this thesis was to study how two anthracyclines, doxorubicin and idarubicin, induced cytotoxicity in tumor cells, as well as their off-target effect on the gastrointestinal system. In Paper I, we exposed four different cancer cell lines to various concentrations of doxorubicin in two different formulations, and measured the cell viability and how much doxorubicin was taken up by the cells. We found that the cell lines differed in their uptake and sensitivity, and that the most resistant cell line had an intracellular exposure of doxorubicin that was about 100 times lower when compared to the more sensitive cell lines. To study the off-target gastrointestinal toxicity, we dosed rats with doxorubicin and studied how chemotherapy-induced mucositis developed during seven days in Paper II. The main effect parameter was intestinal villus atrophy, which was most severe after three days. This was preceded by an increased cell death and decreased proliferation in the crypts, which occurred within the first day after doxorubicin exposure. To study how different cytostatic drugs affected chemotherapy-induced mucositis, and to what extent they caused diarrhea, rats were dosed with one of six different chemotherapies, including doxorubicin and idarubicin, in Paper III. All selected chemotherapies caused similar villus atrophy three days after dosing, but only a third, including idarubicin, caused clear diarrhea. In paper IV the objective was to treat or prevent idarubicin-induced mucositis and diarrhea, by using the anti-inflammatory drugs anakinra and/or dexamethasone. Anakinra alone stopped the diarrhea, while the combination of anakinra and dexamethasone prevented villus atrophy. These positive effects encourage further investigations into the use of anakinra and dexamethasone as supportive th

Background and aim: Transarterial chemoembolization with drug-eluting microspheres (DEM-TACE) is recommended for patients with BCLC stage B hepatocellular carcinoma (HCC) and stage 0-A unsuitable for curative treatments. We assessed efficacy and safety along with hepatobiliary toxicities (HBT) of DEM-TACE using a novel microsphere, LifePearl™, loaded with anthracyclines. Materials and methods: 97 patients diagnosed with HCC were prospectively enrolled and treated using LifePearl™ loaded with doxorubicin (77%) or idarubicin (23%). Safety and tolerability were assessed using CTCAE, HBT by CT/MRI scans, and tumor response by applying modified Response Evaluation Criteria in Solid Tumors (mRECIST). Follow-up was after 2 years. Results: Adverse events (AE) were reported in 73.2% of patients, majority being Grade 1–2. Grade ≥ 3 AE reported in 13.4% of patients were mainly related to postembolization syndrome. HBT were observed after 15.5% (29/187) of the DEM-TACEs. Objective response and disease control rates were 81% and 99%, respectively, as the best responses. Survival rates at one and two years were 81% and 66%, respectively, while the median overall survival (OS) was not reached. Median progression free survival was 13.7 months (95% CI: 11.3; 15.6) and median time to TACE untreatable progression was 16.7 months (95% CI: 12.7; not estimable (n.e.)). Conclusions: DEM-TACE using LifePearl™ provides a high tumor response rate in HCC patients. HBT rates within or below previously reported results for cTACE and DEM-TACE indicate a good safety profile for LifePearl™. The trial was registered in ClinicalTrials.gov National Library of Medicine (ID: NCT03053596)., SCOPUS: ar.j, info:eu-repo/semantics/published

The role of histone deacetylase inhibitors in the treatment of acute myeloid leukemia (AML) is not well characterized. The current study evaluated the safety and efficacy of panobinostat in combination with idarubicin and cytarabine in newly diagnosed patients aged ≤65 years with primary or secondary high-risk AML based on cytogenetic classification. Treatment included fixed dose idarubicin (12 mg/m2/d, IV; day 1–3) and cytarabine (100 mg/m2/d, continuous IV infusion; day 1–7) and escalating oral doses of panobinostat at 15 mg, 20 mg, and 25 mg, thrice weekly starting at week 2 of a 28-day cycle. Forty-six patients were enrolled (primary AML [n = 36], secondary AML [n = 10]). The median age was 55 years. The most common all-grade AEs were diarrhea (54.3%), nausea (39.1%), vomiting, and decreased appetite (each, 21.7%), stomatitis (19.6%), and fatigue (17.4%). The overall response rate was 60.9%, 43.5% achieved a complete remission (CR), and 17.4% achieved CR with incomplete count recovery. The event-free survival at 1-year was 78.3%. Panobinostat in combination with idarubicin and cytarabine demonstrated tolerable safety and efficacy in younger patients with high-risk AML. The recommended phase 2 dose of panobinostat in this combination was 20 mg. ClinicalTrials.gov registry no: NCT01242774, and European Trial Registry EudraCT no: 2009-016809-42.

The purine nucleoside analogues clofarabine and fludarabine are active in acute myeloid leukemia (AML). We conducted a phase I/II randomized study of idarubicin and cytarabine with either clofarabine (CIA) or fludarabine (FIA) for relapsed or refractory AML. Clofarabine 15 mg/m2 was identified as the recommended phase II dose. Eighty-one patients were assigned using adaptive randomization to CIA (n = 48) or FIA (n = 33). The complete response (CR)/CR without platelet recovery rate did not differ between CIA and FIA (38% versus 30%, respectively; p = .50). In both arms, more than half of patients who had received only one prior line of therapy achieved remission. The median event-free survival for CIA and FIA was 2.0 and 1.9 months (p = .48), and the median overall survival was 6.3 and 4.7 months, respectively (p = .28). No significant differences in adverse events or early mortality rates were observed. Overall, CIA and FIA resulted in similar response rates and survival in patients with relapsed/refractory AML.

The purine nucleoside analogues clofarabine and fludarabine are active in acute myeloid leukemia (AML). We conducted a phase I/II randomized study of idarubicin and cytarabine with either clofarabine (CIA) or fludarabine (FIA) for relapsed or refractory AML. Clofarabine 15 mg/m2 was identified as the recommended phase II dose. Eighty-one patients were assigned using adaptive randomization to CIA (n = 48) or FIA (n = 33). The complete response (CR)/CR without platelet recovery rate did not differ between CIA and FIA (38% versus 30%, respectively; p = .50). In both arms, more than half of patients who had received only one prior line of therapy achieved remission. The median event-free survival for CIA and FIA was 2.0 and 1.9 months (p = .48), and the median overall survival was 6.3 and 4.7 months, respectively (p = .28). No significant differences in adverse events or early mortality rates were observed. Overall, CIA and FIA resulted in similar response rates and survival in patients with relapsed/refractory AML.

Acute myeloid leukemia (AML) is a rare yet deadly cancer of the blood and bone marrow. Presently, induction chemotherapy with the DNA damaging drugs cytarabine (ARA-C) and idarubicin (IDA), known as 7 + 3, is the standard of care for most AML patients. However, 7 + 3 is a relatively ineffective therapy, particularly in older patients, and has serious therapy-related toxicities. Therefore, a diagnostic test to predict which patients will respond to 7 + 3 is a critical unmet medical need. We hypothesize that a threshold level of therapy-induced 7 + 3 drug-DNA adducts determines cytotoxicity and clinical response. We further hypothesize that in vitro exposure of AML cells to nontoxic diagnostic microdoses enables prediction of the ability of AML cells to achieve that threshold during treatment. Our test involves dosing cells with very low levels of 14C-labeled drug followed by DNA isolation and quantification of drug-DNA adducts via accelerator mass spectrometry. Here, we have shown proof of principle by correlating ARA-C- and DOX-DNA adduct levels with cellular IC50 values of paired sensitive and resistant cancer cell lines and AML cell lines. Moreover, we have completed a pilot retrospective trial of diagnostic microdosing for 10 viably cryopreserved primary AML samples and observed higher ARA-C- and DOX-DNA adducts in the 7 + 3 responders than nonresponders. These initial results suggest that diagnostic microdosing may be a feasible and useful test for predicting patient response to 7 + 3 induction chemotherapy, leading to improved outcomes for AML patients and reduced treatment-related morbidity and mortality.

Treatment for acute myeloid leukemia (AML) remains suboptimal and many patients remain refractory or relapse upon standard chemotherapy based on nucleoside analogs plus anthracyclines. The crosstalk between AML cells and the BM stroma is a major mechanism underlying therapy resistance in AML. Lenalidomide and pomalidomide, a new generation immunomodulatory drugs (IMiDs), possess pleiotropic anti-leukemic properties including potent immune-modulating effects and are commonly used in hematological malignances associated with intrinsic dysfunctional BM such as myelodysplastic syndromes and multiple myeloma. Whether IMiDs may improve the efficacy of current standard treatment in AML remains understudied. Here, we have exploited in vitro and in vivo preclinical AML models to analyze whether IMiDs potentiate the efficacy of AraC/Idarubicin-based standard AML chemotherapy by interfering with the BM stroma-mediated chemoresistance. We report that IMiDs do not exert cytotoxic effects on either non-del5q/5q- AML cells nor BM-MSCs, but they enhance the immunomodulatory properties of BM-MSCs. When combined with AraC/Idarubicin, IMiDs fail to circumvent BM stromamediated resistance of non-del5q/5q- AML cells in vitro and in vivo but induce robust extramedullary mobilization of AML cells. When administered as a single agent, lenalidomide specifically mobilizes nondel5q/5q- AML cells, but not healthy CD34+ cells, to peripheral blood (PB) through specific downregulation of CXCR4 in AML blasts. Global gene expression profiling supports a migratory/mobilization gene signature in lenalidomide-treated non-del5q/5q- AML blasts but not in CD34+ cells. Collectively, IMiDs mobilize non-del5q/5q- AML blasts to PB through CXCR4 downregulation, but fail to potentiate AraC/Idarubicin activity in preclinical models of non-del5q/5q- AML.

Autofagija, proces programirane ćelijske razgradnje unutarćelijskog sadržaja, je uključena u regulaciju preživljavanja i smrti ćelija kancera. U ovoj doktorskoj disertaciji je po prvi put ispitivana sposobnost antileukemijskih lekova citarabina i idarubicina da indukuju autofagiju u različitim humanim leukemijskim ćelijskim linijama i mononuklearnim ćelijama periferne krvi (MNPK) pacijenata obolelih od leukemije in vitro. Takođe, ispitivani su unutarćelijski mehanizmi odgovorni za indukciju autofagije, kao i uloga autofagije u citotoksičnosti ovih lekova. Vijabilitet REH, HL-60, K562 leukemijskih ćelijskih linija, MNPK pacijenata obolelih od leukemije i MNPK zdravih kontrola je određivan merenjem aktivnosti kisele fosfataze i mitohondrijalnih dehidrogenaza. Protočna citofluorimetrija je korišćena za detekciju apoptoze i zakišeljavanja citoplazme. Indukcija autofagije je ispitivana fluorescentnom mikroskopijom (detekcija unutarćelijskih kiselih vezikula obojenih akridin oranžom), transmisionom elektronskom mikroskopijom (posmatranje autofagnih vezikula) i imunoblot analizom (konverzija LC3-I u LC3-II, degradacija SQSTM1/p62). Aktivacija signalnih puteva koji učestvuju u regulaciji autofagije je analizirana imunoblot metodom. Uloga autofagije u citotoksičnosti citarabina i idarubicina je ispitivana primenom farmakološke inhibicije bafilomicinom A1, hlorokinom, vortmaninom i amonijum hloridom, kao i genetske inaktivacije ekspresije beklina-1, LC3β i SQSTM1 transfekcijom odgovarajućim malim interferirajućim RNK. Citarabin i idarubicin su izazvali povećanje unutarćelijske kiselosti i pojavu autofagnih vezikula sa delimično razgrađenim ćelijskim sadržajem u leukemijskim ćelijskim linijama. Antileukemijski lekovi su stimulisali razgradnju supstrata autofagije SQSTM1 i povećali konverziju LC3-I u LC3-II formu asociranu autofagozomima u odsustvu ili prisustvu inhibitora proteolize, ukazujući tako na povećanje autofagnog fluksa. Oba leka su smanjila fosforilaciju mTOR kinaze, Autophagy, a process of programmed cellular self-digestion, has been implicated in regulation of cancer cell survival and death. The present study investigated for the first time the ability of antileukemic drugs cytarabine and idarubicin to induce autophagy in different human leukemic cell lines and peripheral blood mononuclear cells (PBMC) from leukemia patients in vitro. Intracellular mechanisms responsible for the induction of autophagy, as well as the role of autophagy in cytotoxicity of these drugs were also investigated. Cell viability of REH, HL-60, K562 leukemic cell lines, and PBMC from leukemic patients and healthy controls was determined by measuring the acid phosphatase and mitochondrial succinate dehydrogenase activity. Flow cytometry was used for the detection of apoptosis and intracellular acidification. Autophagy induction was assessed by fluorescent microscopy (detection of acridine orange stained intracellular acidic vesicles), by transmission electron microscopy (observation of autophagic vacuoles), as well as by immunoblot analysis of LC3 conversion and SQSTM1/p62 proteolysis. Activation of autophagy-regulating signaling pathways was analyzed by immunoblotting. Pharmacological inhibition of autophagy with bafilomycin A1, chloroquine, wortmannin, and NH4Cl or RNA interference-mediated knockdown of beclin-1, LC3β and SQSTM1 were used to determine the role of the autophagy in cytotoxicity of antileukemic drugs. Cytarabine and idarubicin induced an increase in intracellular acidification and appearance of autophagic vesicles with partially digested cellular components in leukemic cell lines. Antileukemic drugs stimulated the degradation of autophagic target SQSTM1 and enhanced the conversion of LC3-I to autophagosome-associated LC3-II in the absence or presence of proteolysis inhibitors, thus indicating the increase in autophagic flux...

Autofagija, proces programirane ćelijske razgradnje unutarćelijskog sadržaja, je uključena u regulaciju preživljavanja i smrti ćelija kancera. U ovoj doktorskoj disertaciji je po prvi put ispitivana sposobnost antileukemijskih lekova citarabina i idarubicina da indukuju autofagiju u različitim humanim leukemijskim ćelijskim linijama i mononuklearnim ćelijama periferne krvi (MNPK) pacijenata obolelih od leukemije in vitro. Takođe, ispitivani su unutarćelijski mehanizmi odgovorni za indukciju autofagije, kao i uloga autofagije u citotoksičnosti ovih lekova. Vijabilitet REH, HL-60, K562 leukemijskih ćelijskih linija, MNPK pacijenata obolelih od leukemije i MNPK zdravih kontrola je određivan merenjem aktivnosti kisele fosfataze i mitohondrijalnih dehidrogenaza. Protočna citofluorimetrija je korišćena za detekciju apoptoze i zakišeljavanja citoplazme. Indukcija autofagije je ispitivana fluorescentnom mikroskopijom (detekcija unutarćelijskih kiselih vezikula obojenih akridin oranžom), transmisionom elektronskom mikroskopijom (posmatranje autofagnih vezikula) i imunoblot analizom (konverzija LC3-I u LC3-II, degradacija SQSTM1/p62). Aktivacija signalnih puteva koji učestvuju u regulaciji autofagije je analizirana imunoblot metodom. Uloga autofagije u citotoksičnosti citarabina i idarubicina je ispitivana primenom farmakološke inhibicije bafilomicinom A1, hlorokinom, vortmaninom i amonijum hloridom, kao i genetske inaktivacije ekspresije beklina-1, LC3β i SQSTM1 transfekcijom odgovarajućim malim interferirajućim RNK. Citarabin i idarubicin su izazvali povećanje unutarćelijske kiselosti i pojavu autofagnih vezikula sa delimično razgrađenim ćelijskim sadržajem u leukemijskim ćelijskim linijama. Antileukemijski lekovi su stimulisali razgradnju supstrata autofagije SQSTM1 i povećali konverziju LC3-I u LC3-II formu asociranu autofagozomima u odsustvu ili prisustvu inhibitora proteolize, ukazujući tako na povećanje autofagnog fluksa. Oba leka su smanjila fosforilaciju mTOR kinaze, Autophagy, a process of programmed cellular self-digestion, has been implicated in regulation of cancer cell survival and death. The present study investigated for the first time the ability of antileukemic drugs cytarabine and idarubicin to induce autophagy in different human leukemic cell lines and peripheral blood mononuclear cells (PBMC) from leukemia patients in vitro. Intracellular mechanisms responsible for the induction of autophagy, as well as the role of autophagy in cytotoxicity of these drugs were also investigated. Cell viability of REH, HL-60, K562 leukemic cell lines, and PBMC from leukemic patients and healthy controls was determined by measuring the acid phosphatase and mitochondrial succinate dehydrogenase activity. Flow cytometry was used for the detection of apoptosis and intracellular acidification. Autophagy induction was assessed by fluorescent microscopy (detection of acridine orange stained intracellular acidic vesicles), by transmission electron microscopy (observation of autophagic vacuoles), as well as by immunoblot analysis of LC3 conversion and SQSTM1/p62 proteolysis. Activation of autophagy-regulating signaling pathways was analyzed by immunoblotting. Pharmacological inhibition of autophagy with bafilomycin A1, chloroquine, wortmannin, and NH4Cl or RNA interference-mediated knockdown of beclin-1, LC3β and SQSTM1 were used to determine the role of the autophagy in cytotoxicity of antileukemic drugs. Cytarabine and idarubicin induced an increase in intracellular acidification and appearance of autophagic vesicles with partially digested cellular components in leukemic cell lines. Antileukemic drugs stimulated the degradation of autophagic target SQSTM1 and enhanced the conversion of LC3-I to autophagosome-associated LC3-II in the absence or presence of proteolysis inhibitors, thus indicating the increase in autophagic flux...

BackgroundFludarabine and clofarabine are purine nucleoside analogues with established clinical activity in patients with acute myeloid leukemia (AML).MethodsHerein, the authors evaluated the efficacy and safety of idarubicin and cytarabine with either clofarabine (CIA) or fludarabine (FIA) in adults with newly diagnosed AML. Adults with newly diagnosed AML who were deemed suitable for intensive chemotherapy were randomized using a Bayesian adaptive design to receive CIA (106 patients) or FIA (76 patients). Patients received induction with idarubicin and cytarabine, plus either clofarabine or fludarabine. Responding patients could receive up to 6 cycles of consolidation therapy. Outcomes were compared with a historical cohort of patients who received idarubicin and cytarabine.ResultsThe complete remission/complete remission without platelet recovery rate was similar among patients in the CIA and FIA arms (80% and 82%, respectively). The median event-free survival was 13 months and 12 months, respectively (P = .91), and the median overall survival was 24 months and not reached, respectively (P = .23), in the 2 treatment arms. CIA was associated with more adverse events, particularly transaminase elevation, hyperbilirubinemia, and rash. Early mortality was similar in the 2 arms (60-day mortality rate of 4% for CIA vs 1% for FIA; P = .32). In an exploratory analysis of patients aged <50 years, FIA was found to be associated with improved survival compared with idarubicin and cytarabine (2-year event-free survival rate: 58% vs 30% [P = .05] and 2-year overall survival rate: 72% vs 36% [P = .009]).ConclusionsCIA and FIA have similar efficacy in younger patients with newly diagnosed AML, although FIA is associated with a better toxicity profile. Cancer 2017;123:4430-9. © 2017 American Cancer Society.

BACKGROUND: Fludarabine and clofarabine are purine nucleoside analogues with established clinical activity in patients with acute myeloid leukemia (AML). METHODS: Herein, the authors evaluated the efficacy and safety of idarubicin and cytarabine with either clofarabine (CIA) or fludarabine (FIA) in adults with newly diagnosed AML. Adults with newly diagnosed AML who were deemed suitable for intensive chemotherapy were randomized using a Bayesian adaptive design to receive CIA (106 patients) or FIA (76 patients). Patients received induction with idarubicin and cytarabine, plus either clofarabine or fludarabine. Responding patients could receive up to 6 cycles of consolidation therapy. Outcomes were compared with a historical cohort of patients who received idarubicin and cytarabine. RESULTS: The complete remission/complete remission without platelet recovery rate was similar among patients in the CIA and FIA arms (80% and 82%, respectively). The median event-free survival was 13 months and 12 months, respectively (P = .91), and the median overall survival was 24 months and not reached, respectively (P = .23), in the 2 treatment arms. CIA was associated with more adverse events, particularly transaminase elevation, hyperbilirubinemia, and rash. Early mortality was similar in the 2 arms (60-day mortality rate of 4% for CIA vs 1% for FIA; P = .32). In an exploratory analysis of patients aged <50 years, FIA was found to be associated with improved survival compared with idarubicin and cytarabine (2-year event-free survival rate: 58% vs 30% [P = .05] and 2-year overall survival rate: 72% vs 36% [P = .009]). CONCLUSIONS: CIA and FIA have similar efficacy in younger patients with newly diagnosed AML, although FIA is associated with a better toxicity profile. Cancer 2017;123:4430-9. © 2017 American Cancer Society.

We report the final results of a prospective trial testing the combination of fludarabine, Ara-C and idarubicin (FLAI) followed by low-dose gemtuzumab ozogamicin (FLAI-GO) in 85 patients aged 60 years or more with CD33+ acute myeloid leukaemia (AML). Median age was 68 years (60-82); karyotype was unfavourable in 21 patients (24 %), intermediate in 63 (74 %) and favourable in 1 (2 %). There were five therapy-related deaths. Of the 80 evaluable patients, 47 achieved complete response (CR) (58 %); CR rates were 65 and 32 % in good-intermediate/poor karyotype patients, respectively. Median length of CR was 7 months (3-76). The cumulative incidence of relapse was 84 % with an actuarial survival of 50.3 % at 1 year and 14.4 % at 2 years. The study control population is an unselected consecutive historic cohort of 104 patients treated with the FLAI regimen, who were matched for age and prognostic factors. CR rates after FLAI-GO and FLAI were comparable. However, patients with de novo AML and intermediate-favourable karyotype receiving GO had a significantly lower risk of relapse at 2 years as compared to patients not receiving GO (n = 77) (40 vs 80 %, p = 0.01) and significantly better disease-free survival (p = 0.018) and overall survival (p = 0.022).

All-trans-retinoic acid (ATRA) has greatly modified the prognosis of acute promyelocytic leukemia; however, the role of maintenance in patients in molecular complete remission after consolidation treatment is still debated. From July 1993 to May 2000, 807 genetically proven newly diagnosed acute promyelocytic leukemia patients received ATRA plus idarubicin as induction, followed by 3 intensive consolidation courses. Thereafter, patients reverse-transcribed polymerase chain reaction-negative for the PML-RARA fusion gene were randomized into 4 arms: oral 6-mercaptopurine and intramuscular methotrexate (arm 1); ATRA alone (arm 2); 3 months of arm1 alternating to 15 days of arm 2 (arm 3); and no further therapy (arm 4). Starting from February 1997, randomization was limited to ATRA-containing arms only (arms 2 and 3). Complete remission was achieved in 761 of 807 (94.3%) patients, and 681 completed the consolidation program. Of these, 664 (97.5%) were evaluated for the PML-RARA fusion gene, and 586 of 646 (90.7%) who tested reverse-transcribed polymerase chain reaction-negative were randomized to maintenance. The event-free survival estimate at 12 years was 68.9% (95% confidence interval, 66.4%-71.4%), and no differences in disease-free survival at 12 years were observed among the maintenance arms.

After the identification of discrete relapse-risk categories in patients with acute promyelocytic leukemia (APL) receiving all-trans retinoic and idarubicin (AIDA)-like therapies, the Gruppo Italiano Malattie Ematologiche dell'Adulto (GIMEMA) designed a protocol for newly diagnosed APL (AIDA-2000) in which postremission treatment was risk-adapted. Patients with low/intermediate risk received remission at 3 anthracycline-based consolidation courses, whereas high-risk patients received the same schedule as in the previous, non-risk-adapted AIDA-0493 trial including cytarabine. In addition, all patients in the AIDA-2000 received all-trans retinoic acid (ATRA) for 15 days during each consolidation. After induction, 600 of 636 (94.3%) and 420 of 445 (94.4%) patients achieved complete remission in the AIDA-0493 and AIDA-2000, respectively. The 6-year overall survival and cumulative incidence of relapse (CIR) rates were 78.1% versus 87.4% (P = .001) and 27.7% versus 10.7% (P < .0001). Significantly lower CIR rates for patients in the AIDA-2000 were most evident in the high-risk group (49.7% vs 9.3%, respectively, P < .0001). Our data confirm that anthracycline-based consolidation is at least equally effective as cytarabine-containing regimens for low-/intermediate-risk patients and suggest that a risk-adapted strategy including ATRA for consolidation improves outcome in newly diagnosed APL. Furthermore, our results highlight the role of cytarabine coupled to anthracyclines and ATRA during consolidation in the high-risk group.

Background: The prevalence of and risk factors for central nervous system recurrence in patients with acute promyelocytic leukemia are not well established and remain a controversial matter. Design and Methods: Between 1996 and 2005, 739 patients with newly diagnosed acute promyelocytic leukemia enrolled in two consecutive trials (PETHEMA LPA96 and LPA99) received induction therapy with all-trans retinoic acid and idarubicin. Consolidation therapy comprised three courses of anthracycline monochemotherapy (LPA96), with all-trans retinoic acid and reinforced doses of idarubicin in patients with an intermediate or high risk of relapse (LPA99). Central nervous system prophylaxis was not given. Results: Central nervous system relapse was documented in 11 patients. The 5-year cumulative incidence of central nervous system relapse was 1.7% (LPA96 3.2% and LPA99 1.2%; p=0.09). The cumulative incidence was 0%, 0.8%, and 5.5% in low-, intermediate-, and high-risk patients, respectively. Relapse risk score (p=0.0001) and the occurrence of central nervous system hemorrhage during induction (5-year cumulative incidence 18.7%, p=0.006) were independent risk factors for central nervous system relapse. Conclusions: This study shows a low incidence of central nervous system relapse in patients with acute promyelocytic leukemia following therapy with all-trans retinoic acid and anthracycline without specific central nervous system prophylaxis. Central nervous system relapse was significantly associated with high white blood cell counts and prior central nervous system hemorrhage, which emerged as independent prognostic factors.

Differentiation syndrome (DS) can be a life-threatening complication in patients with acute promyelocytic leukemia (APL) undergoing induction therapy with all- trans retinoic acid (ATRA). Detailed knowl- edge about DS has remained limited. We present an analysis of the incidence, char- acteristics, prognostic factors, and out- come of 739 APL patients treated with ATRA plus idarubicin in 2 consecutive trials (Programa Espanol de Tratamientos en Hematologíc [PETHEMA] LPA96 and LPA99). Overall, 183 patients (24.8%) ex- perienced DS, 93 with a severe form (12.6%) and 90 with a moderate form (12.2%). Severe but not moderate DS was associated with an increase in mortality. A bimodal incidence of DS was observed, with peaks occurring in the first and third weeks after the start of ATRA therapy. A multivariate analysis indicated that a WBC count greater than 5 x 109/L and an abnor- mal serum creatinine level correlated with an increased risk of developing severe DS. Patients receiving systematic pred- nisone prophylaxis (LPA99 trial) in con- trast to those receiving selective prophy- laxis with dexamethasone (LPA96 trial) had a lower incidence of severe DS. Pa- tients developing severe DS showed a reduced 7-year relapse-free survival in the LPA96 trial (60% vs 85%, P = .003), but this difference was not apparent in the LPA99 trial.

The introduction of all-trans retinoic acid (ATRA) and, more recently, arsenic trioxide (ATO) into the therapy of acute promyelocytic leukemia (APL) has revolutionized the management and outcome of this disease. Several treatment strategies using these agents, usually in combination with chemotherapy, but also without or with minimal use of cytotoxic agents, have provided excellent therapeutic results. Cure of APL patients, however, is also dependent on peculiar aspects related to the management and supportive measures that are crucial to counteract life-threatening complications associated with the disease biology and molecularly targeted treatment. The European LeukemiaNet recently appointed an international panel of experts to develop evidence- and expert opinion-based guidelines on the diagnosis and management of APL. Together with providing current indications on genetic diagnosis, modern risk-adapted front-line therapy and salvage treatment, the review contains specific recommendations for the iden

The optimal post-remission treatment for elderly patients with acute myeloid leukemia (AML) is presently unknown. Recent studies have reported the feasibility of autologous peripheral blood stem cell transplantation (PBSCT) in this population. We evaluate the outcome of this post-remission approach after complete remission (CR) and consolidation in elderly patients included in the EORTC-GIMEMA AML-13 trial.

The role of all-trans retinoic acid (ATRA) in pediatric acute promyelocytic leukemia (APL) is the topic of several ongoing studies. The results of the Italian pediatric experience with the multicentric Gruppo Italiano per le Malattie Ematologiche dell'Adulto (GIMEMA)-Italian Pediatric Hematology and Oncology Group (AIEOP) "AIDA" (ATRA and idarubicin) trial are presented. Of the 983 patients with APL enrolled in this protocol between January 1993 and June 2000, 124 (13%) had younger than 18 years. Treatment consisted of ATRA and idarubicin induction followed by 3 polychemotherapy consolidation courses. Molecular response by reverse transcriptase-polymerase chain reaction (RT-PCR) was assessed after consolidation and patients who were PCR- were randomized for different maintenances. One hundred and seven children were eligible and evaluable for induction: 103 (96%) achieved a hematologically complete remission. Overt ATRA syndrome was observed in 2 patients and pseudotumor cerebri was observed in 10 patients. Ninety-four patients were evaluable for RT-PCR analysis at the end of consolidation: 91 (97%) proved PCR+ and 3 PCR-. The overall survival and event-free survival (EFS) are 89% (95% confidence interval [c.i.]: 83%-95%) and 76% (c.i.: 65%-85%), respectively, at more than 10 years. A white blood cell (WBC) count at diagnosis of greater than 10 x 10(9)/L had a significant impact on EFS (59% vs 83% at 10 years). These results highlight the efficacy and feasibility of the AIDA protocol in the pediatric APL population.

A 43 year-old woman presented with fever, abdominal pain, epato-splenomegaly, ascites, cholestasis, anemia, thrombocytopenia and previous diagnosis of sclerosing cholangitis based on liver biopsy and endoscopic retrograde cholangiopancreatography(ERCP). The bone marrow biopsy and the revision of liver biopsy using antitryptase stain diagnosed systemic mastocytosis. Because of the aggressive course of the disease the patient was treated with an acute myeloid leukaemia chemotherapy regimen without success.

Anthracycline antibiotics are widely used as anticancer agents. Idarubicin (4-demethoxydaunorubicin; Ida), a semisynthetic derivative of daunorubicin (Dnr) is more potent than the parent compound in vitro and in vivo. The equitoxic dose of Ida in patients is about one-fourth of that of Dnr. We compared these drugs regarding cytotoxicity, apoptosis induction, and resistance mechanisms in human leukaemic cell lines. Cytotoxicity was studied by means of the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay and drug-induced apoptosis by means of the Annexin V–fluorescein isothiocyanate method at similar intracellular concentrations (extracellular concentrations of 0.35 μM for Ida and 1 μM for Dnr). Ida was at least twice as potent as Dnr in MOLT-4, HL60, CEM, and K562 cell lines. It took 8 h for Ida to induce approximately 20% apoptosis, but at least 22 h for Dnr to reach 20% apoptosis at identical intracellular concentration. Ida induces a faster and higher apoptosis rate compared with Dnr. The human chronic myelogenous leukaemia cell line (K562) was selected for resistance to Dnr and Ida with and without verapamil (Ver). Continuous incubation with Dnr, but not with Ida, led to an increased mdr1 gene expression as assessed by real-time PCR. The development of mdr1 gene expression in Dnr-resistant cells could be reversed by the presence of Ver. Ver also reversed the cytotoxicity to Dnr, but not to Ida, in K562/Dnr cells. The results show that Ida is more effective than Dnr in inducing apoptosis and that there are differences in resistance mechanisms between the drugs.

The outcome of children with acute lymphoblastic leukaemia (ALL) and early relapse remains unsatisfactory. In January 1995, the AIEOP (Associazione Italiana di Oncologia ed Ematologia Pediatrica) group opened a trial for children with ALL in first isolated or combined bone marrow relapse defined at high risk according to the length of first remission and the immunophenotype. The treatment plan included the combination of a single high-dose idarubicin and high-dose cytarabine as induction therapy followed by an intensive consolidation and stem cell transplant (SCT). In total, 100 children from 16 Italian centres were enrolled; 80 out of the 99 evaluable patients (81%) achieved second complete remission; eight (8%) died during induction and 11 (11%) failed to respond. A total of 42 out of the 80 responders (52.5%) received a SCT: 19 from an identical sibling, 11 from a matched unrelated donor and 12 from umbilical cord blood cells. The estimated 4-year overall survival and event-free survival were 25% and 21% respectively. Disease-free survival at 4 years was 25.8% for the 80 responders. At 4 years, 39 out of 100 children remain alive, with 27 of them free of leukaemia. This induction therapy has shown antileukaemic efficacy with acceptable toxicity; moreover, all responders proved eligible for intensive consolidation

The outcome of children with acute lymphoblastic leukaemia (ALL) and early relapse remains unsatisfactory. In January 1995, the AIEOP (Associazione Italiana di Oncologia ed Ematologia Pediatrica) group opened a trial for children with ALL in first isolated or combined bone marrow relapse defined at high risk according to the length of first remission and the immunophenotype. The treatment plan included the combination of a single high-dose idarubicin and high-dose cytarabine as induction therapy followed by an intensive consolidation and stem cell transplant (SCT). In total, 100 children from 16 Italian centres were enrolled; 80 out of the 99 evaluable patients (81%) achieved second complete remission; eight (8%) died during induction and 11 (11%) failed to respond. A total of 42 out of the 80 responders (52.5%) received a SCT: 19 from an identical sibling, 11 from a matched unrelated donor and 12 from umbilical cord blood cells. The estimated 4-year overall survival and event-free survival were 25% and 21% respectively. Disease-free survival at 4 years was 25.8% for the 80 responders. At 4 years, 39 out of 100 children remain alive, with 27 of them free of leukaemia. This induction therapy has shown antileukaemic efficacy with acceptable toxicity; moreover, all responders proved eligible for intensive consolidation.

We investigated the expression of P-glycoprotein (P-gp) in 50 adults with de novo diagnosed acute myeloid leukaemia (AML) and the relationship between presence of P-gp in leukaemic cells and efficacy, as remission induction and survival rate, of two different anthracyclines, daunorubicin (DNR) and idarubicin (IDR). We found that 30 out of 50 patients (60%) were negative (Group 1) and 20 (40%) were positive (Group 2) for P-gp expression evaluated by mean of MRK16 MoAb using a cut-off of 10% positive cells. Thirty-five out of 50 patients (70%) obtained complete remission (CR); depending on P-gp expression, the CR rate was 80% for group 1 and 45% for group 2 (p < 0.005). The median duration of overall survival was 20 months for patients in Group 1 as compared with 10 months for patients of Group 2 (p < 0.005). Regarding the anthracycline used, no significant difference in CR was observed in patients of Group 1 (75% of CR with DNR vs. 90% with IDR); Group 2 obtained 40% of CR with DNR vs. 70% with IDR (p < 0.005). The median duration of overall survival (OS) with the two regimens was comparable in Group 1, while it was significantly longer in patients of Group 2 treated with IDR compared with DNR regimen (p < 0.005). These results confirm the prognostic value of P-gp expression in AML at first appearance and we suggest that idarubicin could be a valid anthracycline drug in the treatment of AML to be evaluated as potential drug of choice in patients with primary or drug-induced multidrug resistance.

To study the effect of the main metabolites on the cytotoxic effect of daunorubicin and idarubicin in human HL-60 cells, drug-sensitive and multidrug-resistant HL60 cells were incubated with idarubicin and daunorubicin and their metabolites idarubicinol and daunorubicinol over a wide range of concentrations. The intracellular uptake of the drugs was determined by photofluorometry, and the cytotoxic effect in vitro was determined by a bioluminescence assay of intracellular adenosine triphosphate (ATP) after 4 days of culture in liquid medium. The effect of intracellular drugs was calculated from the incubation-concentration versus intracellular-uptake and cytotoxic-effect curves. The intracellular uptake of idarubicin was 6 times that of daunorubicin in drug-sensitive cells and 25 times higher in resistant cells. For idarubicinol as compared with daunorubicinol the corresponding factors were 25 and 7, respectively. As compared with the parent substances, the uptake of idarubicinol and daunorubicinol was 16% and 4%, respectively, in sensitive cells and 40% and >100%, respectively, in resistant cells. An intracellular concentration of 0.5 nmol/mg protein of both parent substances caused a 50% growth inhibition in drug-sensitive cells as compared with 10 nmol/mg protein for drug-resistant cells. For the metabolites an intracellular concentration of 0.4 nmol/mg protein of idarubicinol and 2.0 nmol/mg protein of daunorubicinol was required to inhibit cells’ growth by 50% in drug-sensitive HL60 cells. In the resistant HL60 cells the corresponding values were 30 nmol/mg protein for idarubicinol and 40 nmol/mg protein for daunorubicinol. These results confirm that idarubicinol may significantly contribute to the clinical effect of idarubicin. However, in combination with previous results that have shown low intracellular concentrations of the metabolites in vivo, it appears that the pharmacokinetic properties of the mother substances provide the major explanation for the

We investigated the expression of P-glycoprotein (P-gp) in 50 adults with de novo acute myeloid leukemia (AML) at the initial diagnosis in order to further define the relationship between the presence of P-gp on leukemic cells and the efficacy of two different anthracycline drugs, Daunorubicin (DNR) and Idarubicin (IRR), in terms of remission, induction and survival. We found that 30 (60%) of the 50 patients were negative for P-gp expression (group 1) and 20 patients (40%) were positive (group 2) for P-gp expression by MRK16 MoAb using a cut of 10% positive cells. Among the 50 patients, 35 (70%) obtained complete remission (CR); depending on P-gp expression the CR rate was 80% for group 1 and 45% for group 2 (p<0.005). The median duration of overall survival (OS) was 20 months for patients in group 1, compared to 10 months for patients in group 2 (p<0.005). Regarding the anthracycline used, no difference in CR has been observed in patients of group 1 (75% CTR with DNR versus 90% CR with IDR); on the contrary in group 2 we observed 40% CR with DNR versus 70% CR with IDR (p<0.005). No significant difference has been achieved in group 1 terms of median duration of overall survival between DNR and IDR regimen; on the contrary the median duration of OS in patients of group 2 treated with IDR regimen was significantly longer than DNR regimen (p<0.005). These results confirm the prognostic value of P-gp expression in AML at diagnosis and we suggest that Idarubicin could be a valid anthracycline drug for reversing multidrug resistance.