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1.
Leuk Lymphoma ; 42(1-2): 89-98, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11699226

ABSTRACT

We treated 33 patients with a variant of the standard 3 weekly CHOP regime, replacing doxorubicin with liposomal daunorubicin (DaunoXome, NeXstar Pharmaceuticals) 120 mg/m2 (COP-X). Eighteen subjects had relapsed/refractory aggressive NHL and 15 had indolent NHL/CLL. Median number of courses received was 4 (1-8). Thirty-two patients were evaluable for efficacy and 26 (81%) responded. 88% of patients with aggressive NHL responded; three (18%) patients achieved complete remission (CR), 12 (70%) achieved partial remission (PR), 1 (6%) patient had stable disease (SD) and 1 (6%) patient progressed through treatment. Median duration of response for patients with aggressive NHL was 3 months. The response rate in indolent NHL/CLL was 73%. Four (27%) patients achieved CR, 7 (46%) PR and 4 (27%) SD. At two years post treatment, 55% of the patients with indolent NHL/CLL remain progression-free, although 4 patients have proceeded to consolidation therapy. Twenty-seven out of 28 (96%) patients developed neutropenia of short duration following one or more of their treatments. Twenty-three patients developed an infection at some stage during therapy (all associated with neutropenia) and required hospitalisation. There were two toxic deaths (infection) both of which occurred in patients who were neutropenic before starting COP-X. Platelet toxicity was mild in patients with normal platelet counts at the commencement of therapy. Alopecia and mucositis were mild. No clinical evidence of myocardial failure was observed. We conclude that the substitution of DaunoXome for doxorubicin in the CHOP regimen to form COP-X provides excellent efficacy against non-Hodgkin's lymphoma. Response durations were short but comparable to those reported with other regimens. COP-X was well tolerated with some suggestion of reduced non-haematological toxicity. The regimen should be considered as an alternative to CHOP with potentially less non-haematological toxicity, particularly cardiac; further studies are required to evaluate the regimen in this context.


Subject(s)
Antibiotics, Antineoplastic/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Daunorubicin/administration & dosage , Lymphoma, Non-Hodgkin/drug therapy , Salvage Therapy/methods , Aged , Antibiotics, Antineoplastic/toxicity , Antineoplastic Combined Chemotherapy Protocols/toxicity , Cyclophosphamide/administration & dosage , Daunorubicin/toxicity , Female , Humans , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , Liposomes/administration & dosage , Male , Middle Aged , Prednisolone/administration & dosage , Prognosis , Remission Induction , Survival Analysis , Therapeutic Equivalency , Treatment Outcome , Vincristine/administration & dosage
2.
Biochem Biophys Res Commun ; 287(1): 181-9, 2001 Sep 14.
Article in English | MEDLINE | ID: mdl-11549272

ABSTRACT

Tumor necrosis factor-alpha (TNFalpha) mediates cytochrome c release from mitochondria, loss of mitochondrial membrane potential (DeltaPsim) and apoptosis in sensitive leukemic cells. In the present study, by using the human leukemic U937 cell line, we demonstrate that the cytochrome c release is caspase-8-dependent and can be blocked by an inhibitor of caspase-8, Z-Ile-Glu (OMe)-Thr-Asp(OMe)-fluoromethyl ketone (Z-IETD.fmk), or a pan caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD.fmk). However, TNFalpha-mediated loss of DeltaPsim was not inhibited by caspase inhibitors. The apoptotic process was blocked by either Z-IETD.fmk or Z-VAD.fmk in cells with lower DeltaPsim. U937 cells with stable transfection of the cellular inhibitor of apoptosis protein 1 (c-IAP1) are resistant to TNFalpha-induced activation of caspases, Bid cleavage, cytochrome c release and DeltaPsim collapse. In addition, both c-IAP1 and XIAP were not up-regulated upon prolonged exposure to TNFalpha. In contrast, there was a caspase-dependent cleavage of XIAP, but not c-IAP1, during treatment with TNFalpha for 7 days. These results demonstrate that c-IAP1 blocks TNFalpha signaling at a level controlling both activation of caspase-8 and a signal to cause loss of DeltaPsim. The sensitive U937 cell line failed to acquire resistance and gain a self-protecting advantage against apoptosis, upon induction of c-IAP1 expression.


Subject(s)
Apoptosis , Caspases/metabolism , Mitochondria/drug effects , Tumor Necrosis Factor-alpha/pharmacology , Viral Proteins/pharmacology , BH3 Interacting Domain Death Agonist Protein , Carrier Proteins/metabolism , Caspase 3 , Caspase 8 , Caspase 9 , Caspase Inhibitors , Cytochrome c Group/metabolism , Drug Interactions , Enzyme Activation/drug effects , Enzyme Inhibitors/pharmacology , Humans , Inhibitor of Apoptosis Proteins , Leukemia/pathology , Membrane Potentials/drug effects , Mitochondria/enzymology , Mitochondria/physiology , Proteins/metabolism , Signal Transduction/drug effects , Transfection , Tumor Cells, Cultured , U937 Cells , Ultraviolet Rays , Viral Proteins/genetics , X-Linked Inhibitor of Apoptosis Protein
3.
Oncogene ; 20(35): 4817-26, 2001 Aug 09.
Article in English | MEDLINE | ID: mdl-11521193

ABSTRACT

Bax translocation from cytosol to mitochondria is believed to be a crucial step for triggering cytochrome c release from mitochondria. However, it is unclear whether Bax translocation is associated with Bax induction by DNA damaging agents. The induction of Bax in response to DNA damaging agents has been considered to be linked with p53. In this study, we used the p53 negative human chronic myeloid leukaemia K562 cell line. Bax up-regulation occurred at the whole cell level after DNA damage induced by etoposide. However, after incubation with etoposide, Bax failed to translocate to mitochondria and as a result, the apoptotic process was blocked. A Bax stable transfectant, the K/Bax cell line, expressed more Bax protein in the cytosol, mitochondria and nuclei. This Bax overexpression induced cytochrome c release, a reduction of cytochrome c oxidase activity and mitochondrial membrane potential (Delta(Psi)m). However, Bax-induced apoptosis was blocked downstream of mitochondria in K562 cells. The increased levels of mitochondrial Bax sensitized cells to etoposide-induced activation of caspases-2, -3 and -9 and apoptosis. However, after transient transfection with the Apaf-1 gene, K/Bax cells were sensitized to etoposide-induced caspase activation and apoptosis to a larger extent compared with Bax or Apaf-1 transfection alone. We therefore conclude that two mechanisms contribute to the resistance of K562 cells to etoposide-induced apoptosis; firstly failure of Bax targeting to mitochondria and, secondly, deficiency of Apaf-1. Uncoupling of Bax translocation from Bax induction can occur in response to etoposide-induced DNA damage.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Etoposide/pharmacology , Leukemia/drug therapy , Proto-Oncogene Proteins c-bcl-2 , Proto-Oncogene Proteins/metabolism , Apoptotic Protease-Activating Factor 1 , Biological Transport , Caspases/physiology , DNA Damage , Humans , K562 Cells , Leukemia/pathology , Proteins/physiology , bcl-2-Associated X Protein
4.
Apoptosis ; 6(4): 269-78, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11445669

ABSTRACT

Quantitation of apoptotic cell death in vivo has become an important issue for patients with acute leukemia. We describe herein a new analytical method, based on infrared (IR) spectroscopy, to estimate the percentage of apoptotic leukemic cells in two different cell lines (CEM and K562), induced with etoposide (VP-16). As the percentage of apoptosis increases, the protein structure shifts from dominantly beta-sheet to unordered (random coil), the overall lipid content increases and the amount of detectable DNA decreases. These changes can be directly related to the percentage of apoptosis as determined by two standard reference methods: flow cytometry and DNA ladder formation. The correlation between the significant IR spectral changes and the percentage of apoptotic leukemia cells in the two cell lines was optimal up to 24 h following etoposide treatment (r = 0.99 for CEM cells and r = 0.96 for K562 cells). Furthermore, IR spectroscopy is able to detect apoptotic changes in these cells already after 4 h treatment with VP-16, compared to flow cytometry which needs 6 h to observe significant changes. Our study suggests that IR spectroscopy may have potential clinical utility for the early, fast and reagent free assessment of chemotherapeutic efficacy in patients with leukemia.


Subject(s)
Apoptosis , Leukemia/pathology , Spectrophotometry, Infrared , Cell Count , Etoposide/pharmacology , Flow Cytometry , Tumor Cells, Cultured
5.
Blood ; 98(2): 414-21, 2001 Jul 15.
Article in English | MEDLINE | ID: mdl-11435311

ABSTRACT

The human leukemia cell lines K562, CEM, CEM/VLB(100), human leukemic blasts, and the bladder cancer J82 cell line have different sensitivities to UV light-induced apoptosis. It is reported that resistance to UV light-induced apoptosis occurs at a point in the apoptotic pathway upstream of caspase-3 but downstream of mitochondrial cytochrome c release. It is demonstrated that the block is due to deficiency of Apaf-1, a critical member of the apoptosome. Sensitivity to apoptosis was independent of caspase-9b or XIAP (inhibitors of apoptosis proteins) expression or levels of procaspase-9. Transfection of Apaf-1 conferred sensitivity to apoptosis in resistant cells. Apaf-1 deficiency may constitute a significant mode of resistance to apoptosis in human leukemia.


Subject(s)
Apoptosis , Cytochrome c Group/metabolism , Proteins/analysis , Antibodies/pharmacology , Apoptosis/drug effects , Apoptotic Protease-Activating Factor 1 , Caspase 3 , Caspase 9 , Caspases/analysis , Caspases/metabolism , Cytochrome c Group/pharmacology , Enzyme Activation/drug effects , Granzymes , Humans , Leukemia, Myeloid, Acute , Leukemia-Lymphoma, Adult T-Cell , Protein Precursors/metabolism , Proteins/genetics , Proteins/immunology , Proteins/metabolism , Serine Endopeptidases/pharmacology , Transfection , Tumor Cells, Cultured , Ultraviolet Rays , Urinary Bladder Neoplasms , X-Linked Inhibitor of Apoptosis Protein
6.
Biochem Biophys Res Commun ; 283(5): 1037-45, 2001 May 25.
Article in English | MEDLINE | ID: mdl-11355877

ABSTRACT

We have previously shown that Bax translocation was crucial in TNFalpha or etoposide-induced apoptosis. Overexpression of Bax sensitized chronic myeloid leukemic K562 cells to etoposide-induced apoptosis. Treatment with TNF-related apoptosis-inducing ligand (TRAIL) induces a loss of mitochondrial membrane potential (DeltaPsim), cytochrome c release from mitochondria, activation of caspases-8, -9, and -3, and cleavage of Bid in the K562 cell line. Bax failed to sensitize K562 cells to TRAIL-induced apoptosis. TRAIL did not induce Bax expression and/or translocation from cytosol to mitochondria in the K562 cell line. However, 100 microM Z-VAD.fmk, a pan caspase inhibitor, completely blocked TRAIL-initiated mitochondrial alterations and cleavages of caspases and Bid. We propose that TRAIL-induced apoptosis in K562 cells is via Type I apoptotic signal pathway. Bax translocation is not essential for TRAIL-induced cytochrome c release and DeltaPsim collapse in the Type I cells.


Subject(s)
Apoptosis/physiology , Membrane Glycoproteins/pharmacology , Proto-Oncogene Proteins c-bcl-2 , Tumor Necrosis Factor-alpha/pharmacology , Amino Acid Chloromethyl Ketones/pharmacology , Apoptosis/drug effects , Apoptosis Regulatory Proteins , BH3 Interacting Domain Death Agonist Protein , Carrier Proteins/metabolism , Caspase 3 , Caspase 8 , Caspase 9 , Caspases/metabolism , Cysteine Proteinase Inhibitors/pharmacology , Cytochrome c Group/metabolism , Enzyme Precursors/metabolism , Humans , Intracellular Membranes/physiology , K562 Cells , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Membrane Potentials/drug effects , Membrane Potentials/physiology , Mitochondria/physiology , Proto-Oncogene Proteins/genetics , Recombinant Proteins/metabolism , TNF-Related Apoptosis-Inducing Ligand , Transfection , bcl-2-Associated X Protein
7.
Leuk Res ; 25(5): 423-31, 2001 May.
Article in English | MEDLINE | ID: mdl-11301111

ABSTRACT

These data show that 8-Cl-cAMP is cytotoxic to the lymphoblastic leukemia cell line CEM and its vinblastine selected multidrug resistant derivative, CEM/VLB100 although PKA was not involved in these effects. The cytotoxic effects of 8-Cl-cAMP was abrogated by cotreatment with either ADA or IBMX which indicated a degradation form of 8-Cl-cAMP was needed for this cytotoxicity. CEM and CEM/VLB100 cells displayed a notable sensitivity to 8-Cl-adenosine-induced growth inhibition and apoptosis. 8-Cl-adenosine increased the cytosolic levels of IkappaBalpha which prevented NF-kappaB nuclear translocation. 8-Cl-adenosine also prevented TNFalpha-induced IkB decay and NF-kappaB activation in CEM and CEM/VLB100 cells.


Subject(s)
2-Chloroadenosine/analogs & derivatives , 2-Chloroadenosine/pharmacology , 8-Bromo Cyclic Adenosine Monophosphate/analogs & derivatives , Antineoplastic Agents/pharmacology , Antineoplastic Combined Chemotherapy Protocols/pharmacology , Apoptosis/drug effects , I-kappa B Proteins , Leukemia, T-Cell/drug therapy , NF-kappa B/physiology , Tumor Necrosis Factor-alpha/pharmacology , 2-Chloroadenosine/administration & dosage , 2-Chloroadenosine/pharmacokinetics , 8-Bromo Cyclic Adenosine Monophosphate/administration & dosage , 8-Bromo Cyclic Adenosine Monophosphate/pharmacokinetics , 8-Bromo Cyclic Adenosine Monophosphate/pharmacology , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/pharmacokinetics , Antineoplastic Agents, Phytogenic/pharmacology , Biotransformation , Cell Division/drug effects , Cell Nucleus/metabolism , DNA-Binding Proteins/metabolism , Dose-Response Relationship, Drug , Drug Resistance, Neoplasm , Growth Inhibitors/pharmacology , Humans , Leukemia, T-Cell/metabolism , Leukemia, T-Cell/pathology , NF-KappaB Inhibitor alpha , NF-kappa B/biosynthesis , NF-kappa B/metabolism , Tumor Necrosis Factor-alpha/administration & dosage , Up-Regulation/drug effects , Vinblastine/pharmacology
8.
Biochem Biophys Res Commun ; 282(1): 268-72, 2001 Mar 23.
Article in English | MEDLINE | ID: mdl-11264002

ABSTRACT

Apaf-1 plays a crucial role in the cytochrome c/dATP-dependent activation of caspase-9 and -3. We found that the human myeloid leukemic K562 cells were more resistant to cytochrome c-induced activation of caspase-9 and -3 in a cell-free system compared with the human T-lymphoblastic subclone CEM/VLB(100) cells. Apaf-1 cDNA sequencing revealed an additional insert of 11 aa between the CARD and CED-4 (ATPase) domains in K562 cells, which was identical to the sequence of Apaf-1XL. Immunoprecipitation of Apaf-1 with caspase-9 after a cell-free reaction demonstrated that Apaf-1XL in the K562 cell line showed a lower binding ability to caspase-9 compared with Apaf-1L protein. The resistance of K562 cells to cytochrome c-dependent apoptosis may be partly due to this Apaf-1XL form. These results suggest that the additional insert between CARD and CED-4 domains might affect Apaf-1 recruitment of caspase-9 during apoptosis.


Subject(s)
Protein Isoforms/metabolism , Proteins/metabolism , Adenosine Triphosphatases/metabolism , Apoptotic Protease-Activating Factor 1 , Caspase 3 , Caspase 9 , Caspases/metabolism , Cytochrome c Group/metabolism , DNA, Complementary , Enzyme Activation , Humans , Protein Isoforms/chemistry , Protein Isoforms/genetics , Proteins/chemistry , Proteins/genetics , Tumor Cells, Cultured
9.
Br J Haematol ; 115(4): 817-25, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11843815

ABSTRACT

The anthracycline antibiotic idarubicin (IDA) induces double-stranded DNA breaks, the generation of reactive oxygen species (ROS) and apoptosis in human leukaemic cells. It is unclear whether the generation of ROS is associated with the apoptotic process. Using the T-lymphoblastic leukaemic CEM cell line, we found that IDA-induced DNA breaks were correlated with final cell death. The reduction in mitochondrial membrane potential (Deltapsim) and the generation of ROS occurred simultaneously with IDA-induced activation of caspase-9 and caspase-3. Inhibition of caspases by a pan-caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-fmk) completely blocked IDA-induced reduction of Deltapsim, apoptosis and final cell death. Interestingly, ROS generation was significantly enhanced by Z-VAD-fmk. ROS generation was neither caspase dependent nor part of the apoptotic process. IDA-mediated reduction in Deltapsim is caspase dependent and is not a consequence of the generation of ROS. These results indicate that IDA-induced generation of ROS and apoptosis are separate events. Inhibition of caspases facilitates IDA-mediated generation of ROS.


Subject(s)
Antibiotics, Antineoplastic/adverse effects , Apoptosis , DNA Damage , Idarubicin/adverse effects , Leukemia-Lymphoma, Adult T-Cell/metabolism , Reactive Oxygen Species/metabolism , Amino Acid Chloromethyl Ketones/pharmacology , Caspase 3 , Caspase 9 , Caspase Inhibitors , Caspases/metabolism , Cell Death , Cell Line , Enzyme Activation , Enzyme Inhibitors/pharmacology , Humans , Membrane Potentials/drug effects , Mitochondria/physiology
10.
Med Oncol ; 17(4): 333-6, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11114714

ABSTRACT

A 36-year-old man underwent matched unrelated donor bone marrow transplantation for chronic myeloid leukaemia. He developed severe hepatic veno-occlusive disease as an early post-transplant complication. Tissue plasminogen activator was initially felt to be contraindicated since the patient had concomitant pericarditis. Defibrotide was therefore commenced as treatment for veno-occlusive disease. The pericarditis improved but the veno-occlusive disease continued to worsen (peak bilirubin 353 micromol/l). Tissue plasminogen activator followed by a heparin infusion was therefore administered. However, he proceeded to develop haemorrhagic cardiac tamponade that required drainage. Thrombolysis was therefore discontinued and treatment with defibrotide resumed after an interval of 48 h. The veno-occlusive disease gradually resolved and defibrotide was discontinued once the bilirubin had plateaued. He was discharged home on day +52 post-transplant.


Subject(s)
Bone Marrow Transplantation/adverse effects , Fibrinolytic Agents/therapeutic use , Hepatic Veno-Occlusive Disease/drug therapy , Hepatic Veno-Occlusive Disease/etiology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/therapy , Polydeoxyribonucleotides/therapeutic use , Tissue Plasminogen Activator/therapeutic use , Adult , Cardiac Tamponade , Humans , Male , Pericarditis , Transplantation, Homologous , Treatment Outcome
11.
Br J Haematol ; 111(3): 807-16, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11122142

ABSTRACT

The signal transduction pathway through which tumour necrosis factor (TNF) induces apoptosis in leukaemic cells may involve activation of cytosolic phospholipase A(2) (cPLA(2)). The steroids dexamethasone (Dex) and 1,25(OH)(2) D(3) both render U937 leukaemic cells resistant to TNF-induced apoptosis. In this study, we found that Dex inhibited both spontaneous and TNF-induced activation of cPLA(2). Dex had no direct effect on cellular cPLA(2) levels, but facilitated cPLA(2) degradation upon subsequent stimulation of cells with TNF. In addition, Dex increased synthesis of the endogenous cPLA(2) inhibitor lipocortin 1 (LC1). An antisense oligonucleotide to LC1 could completely abrogate Dex-induced resistance to the cytotoxic action of TNF. Constitutive LC1 levels were relatively higher in myeloid leukaemic blasts showing resistance to TNF than TNF-sensitive myeloid leukaemic cell lines. Our data suggest that Dex confers the resistance of U937 cells to TNF-induced apoptosis by upregulating intracellular levels of LC1 and by facilitating a negative-feedback loop, which is activated upon stimulation with TNF. High constitutive levels of LC1 in leukaemic blasts may protect them against immune-mediated killing.


Subject(s)
Annexin A1/metabolism , Antineoplastic Agents, Hormonal/pharmacology , Dexamethasone/pharmacology , Leukemia/metabolism , Signal Transduction/drug effects , Tumor Necrosis Factor-alpha/pharmacology , Acute Disease , Annexin A1/genetics , Apoptosis/drug effects , Arachidonic Acid/metabolism , Calcimycin/pharmacology , Cell Line , Cytosol/enzymology , Enzyme Activation/drug effects , Enzyme Inhibitors/metabolism , Feedback , Humans , Ionophores/pharmacology , Leukemia, Monocytic, Acute/metabolism , Leukemia, Myeloid/metabolism , Oligonucleotides, Antisense/pharmacology , Phospholipases A/antagonists & inhibitors , Phospholipases A/metabolism , Stimulation, Chemical
12.
Ann Oncol ; 11(7): 857-60, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10997814

ABSTRACT

BACKGROUND: Although lymphoid malignancies are generally chemosensitive, relapse is common. The use of high-dose therapy can make subsequent cytotoxic therapy intolerable. There is a need to develop regimens with low acute toxicity which are suitable for use in patients post-high dose therapy and following the failure of standard protocols. PATIENTS AND METHODS: Twenty-six patients with lymphomas, fifteen of whom had received high-dose therapy, were treated with a novel regimen consisting of low-dose lomustine, chlorambucil, daily subcutaneous bleomycin, vincristine and methotrexate with dexamethasone on an eight-week cycle (LBCMVD-56). A median of three cycles was given. RESULTS: The overall response rate at 12 weeks was 67% (21% complete remission (CR)) with a median overall survival of 13 months. A symptomatic response was seen in 72%. Previous high-dose therapy did not compromise the response rate. Toxicity was acceptable with grade 3-4 haematological toxicity seen in 27% of cycles, gastrointestinal toxicity seen in 11% and pulmonary toxicity seen in 8%. Thirty-one percent of patients required hospitalisation at some point during this treatment most commonly for neutropenic sepsis. CONCLUSIONS: LBCMVD-56 is an inexpensive, outpatient-based regimen with low acute toxicity and a high response rate in this heavily pre-treated group of patients.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Hodgkin Disease/drug therapy , Lymphoma, Non-Hodgkin/drug therapy , Adolescent , Adult , Aged , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Bleomycin/administration & dosage , Chlorambucil/administration & dosage , Dexamethasone/administration & dosage , Drug Administration Schedule , Female , Hodgkin Disease/pathology , Hospitalization , Humans , Infusions, Intravenous , Injections, Subcutaneous , Lomustine/administration & dosage , Lymphoma, Non-Hodgkin/pathology , Male , Methotrexate/administration & dosage , Middle Aged , Neutropenia/chemically induced , Sepsis/chemically induced , Treatment Outcome , Vincristine/administration & dosage
13.
Br J Haematol ; 109(3): 571-5, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10886206

ABSTRACT

We report the results of a non-randomized pilot study of an oral regimen comprising CCNU (lomustine; 25 or 50 mg/m2 on day 1), idarubicin (4-demethoxydaunorubicin) (10 mg/m2 on days 1-3) and dexamethasone (10 mg b.d. on days 1-4) in patients with relapsed or refractory myeloma. Treatment was given every 28 d for a maximum of six courses. Sixty patients were entered of whom 57 were evaluable. Overall response rate (partial or minor response) was 49% with 30% of patients achieving a partial response (50% tumour reduction). Response rates were higher in patients with untested relapse than in those with refractory disease (overall response rates 56% vs. 31%). The major toxicity was neutropenia and the regimen was otherwise well tolerated. The median survival from entry of all patients was 15 months, with 30% of patients alive at 2 years. This regimen represents a useful addition to available treatment options.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Multiple Myeloma/drug therapy , Adult , Aged , Aged, 80 and over , Antibiotics, Antineoplastic/administration & dosage , Antibiotics, Antineoplastic/adverse effects , Antineoplastic Agents, Alkylating/administration & dosage , Antineoplastic Agents, Alkylating/adverse effects , Antineoplastic Agents, Hormonal/administration & dosage , Antineoplastic Agents, Hormonal/adverse effects , Antineoplastic Combined Chemotherapy Protocols/adverse effects , Dexamethasone/administration & dosage , Dexamethasone/adverse effects , Female , Humans , Idarubicin/administration & dosage , Idarubicin/adverse effects , Lomustine/administration & dosage , Lomustine/adverse effects , Male , Middle Aged , Multiple Myeloma/mortality , Neutropenia/chemically induced , Pilot Projects , Recurrence , Survival Rate
14.
Blood Rev ; 14(1): 14-30, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10805258

ABSTRACT

Pooled human intravenous immunoglobulin (IVIg) has been used successfully to treat or ameliorate the clinical manifestations of humoral immune deficiencies, haematological disorders, HIV infection and many other diseases states. However, the mechanism of action of IVIg remains unclear. Several mechanisms of action of IVIg have been proposed. These include Fcy receptor blockade, accelerated clearance of endogenous pathogenic auto-antibodies, inhibition of components of the complement cascade, neutralization of super-antigens and bacterial toxins as well as anti-cytokine and anti-idiotype effects. A major contributor to host immunity and immune surveillance against infection, tissue or cell damage and malignancy is the monocyte/macrophage system. Monocyte-directed inflammation is a desirable consequence of microbiological or malignant challenge. However, monocyte hyperactivity may contribute to certain pathological conditions. These include the systemic inflammatory response syndrome (SIRS), septic shock, other dysregulated inflammatory disorders and auto-immunity. Novel therapies that can suppress the hyperactive state or correct monocyte/macrophage dysfunction without compromising normal host cell-mediated immunity are desirable. In this review, we discuss the immunomodulatory effects of IVIg focussing particularly upon the monocyte/macrophage system in pertinent disease states.


Subject(s)
Adjuvants, Immunologic/therapeutic use , Immunoglobulins, Intravenous/therapeutic use , Macrophages/immunology , Monocytes/immunology , Adjuvants, Immunologic/chemistry , Animals , Humans , Immunoglobulins, Intravenous/chemistry , Immunologic Deficiency Syndromes/therapy
15.
Leukemia ; 14(4): 620-8, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10764147

ABSTRACT

Alterations in the response of leukaemic cells to apoptosis-inducing stimuli may account for resistance to chemotherapy and treatment failure, either by disruption of the apoptotic pathway itself or by altered DNA repair; quiescent cells and those with disrupted cell-cycle checkpoints may also display decreased apoptosis. Quiescence can be induced by the differentiation of myeloid cells, and this led us to investigate whether the modulation of drug-induced apoptosis associated with differentiation might be a model for quiescence-associated resistance generally. We have demonstrated that resistance to idarubicin-induced apoptosis increased with greater duration of incubation of HL60 and U937 cells with ATRA and 1,25(OH)2 D3 and that this protective effect correlated with the degree of G0/G1 accumulation. In addition, the cytoprotective effects held for other classes of cytotoxic drugs with different mechanisms of action to idarubicin. Prolonged exposure to idarubicin or vinblastine was associated with diminution of the protective effect and re-entry of cells into cycle. The full cytoprotective effect was restored by resupplementation with ATRA or 1,25(OH)2 D3 during exposure to idarubicin, with concomitant persistence of G0/G1 accumulation. Differentiating agents prevented the accumulation of leukaemic cells at the G2/M checkpoint in response to low concentrations of idarubicin. Understanding how differentiating agents modulate these cell-cycle checkpoints, and how quiescent cells evade apoptosis, may allow the development of therapeutic strategies to limit such apoptosis-inhibiting effects and maximise cell kill from chemotherapy.


Subject(s)
Apoptosis/drug effects , Cell Cycle/drug effects , Drug Resistance, Neoplasm/physiology , Idarubicin/pharmacology , Leukemia, Myeloid/pathology , Neoplastic Stem Cells/drug effects , Vinblastine/pharmacology , Acute Disease , Calcitriol/pharmacology , Cell Differentiation/drug effects , Dose-Response Relationship, Drug , G1 Phase/drug effects , HL-60 Cells/drug effects , HL-60 Cells/pathology , Humans , Neoplastic Stem Cells/pathology , Resting Phase, Cell Cycle/drug effects , Tretinoin/pharmacology , U937 Cells/drug effects , U937 Cells/pathology
16.
Br J Haematol ; 108(3): 565-73, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10759715

ABSTRACT

The cyclic adenosine monophosphate (cAMP)-dependent protein kinase (PKA) signal pathway regulates cell proliferation, differentiation and cell death. It may also regulate the multidrug resistance (MDR) phenotype in leukaemic cells. These data showed that MDR1+ K/Dau600 cells exhibited a higher basal level of PKA activity than MDR- parental cells. The significance of this on tumour necrosis factor alpha (TNFalpha)-induced apoptosis and cytostasis was investigated further. In comparison with MDR1- parental cells, K/Dau600 cells had a higher expression of PKA regulatory subunit RIalpha and nuclear catalytic subunit PKAcalpha. They were also more susceptible to inhibition of proliferation and induction of apoptosis by TNFalpha and/or forskolin, but this could be attenuated by H89. An increase in cAMP was associated with the apoptosis in the K/Dau600 cell line. Forskolin inactivated NF-kappaB in K/Dau600 cells but not in K562 cl. 6 cells, whereas TNF activated NF-kappaB in K562 cl.6 cells but not in K/Dau600 cells. 8-Cl-cAMP exhibited similar inhibitory effects on the proliferation of all of the cell lines used via its metabolite 8-Cl-adenosine, which indicates that these effects were independent of residual PKA or cAMP. Therefore, the differential sensitivity to apoptosis and/or growth inhibition could be mediated via cAMP, partly through PKA via NF-kappaB and partly by PKA-independent pathways.


Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , Antineoplastic Agents/therapeutic use , Apoptosis/drug effects , Colforsin/therapeutic use , Cyclic AMP-Dependent Protein Kinases/metabolism , Leukemia/enzymology , Sulfonamides , 8-Bromo Cyclic Adenosine Monophosphate/analogs & derivatives , 8-Bromo Cyclic Adenosine Monophosphate/pharmacology , Cell Division/drug effects , Cyclic AMP/metabolism , Cyclic AMP-Dependent Protein Kinases/antagonists & inhibitors , Enzyme Inhibitors/pharmacology , Humans , Isoquinolines/pharmacology , Leukemia/drug therapy , Leukemia/metabolism , NF-kappa B/metabolism , Tumor Cells, Cultured , Tumor Necrosis Factor-alpha/therapeutic use
17.
Leuk Res ; 24(4): 317-30, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10713329

ABSTRACT

Cytokines such as interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) activate monocytes both in vitro and in vivo. We therefore studied whether the anti-leukaemic activity of monocytes could be augmented by IL-3 alone or in combination with GM-CSF. Using normal human monocytes stimulated with IL-3, GM-CSF, LPS or combinations of growth factor and LPS, we studied their cytotoxic activity against leukaemic cell-lines and primary AML blasts. IL-3 like GM-CSF, augmented the expression and secretion of TNF but did not prime for further expression and secretion of TNF in response to LPS. Neither GM-CSF or IL-3 increased the expression or secretion of TNF receptor p55 (TNF-Rp55), although both agents increased expression of TNF receptor p75 (TNF-Rp75). Monocyte-mediated cytotoxicity (MMC) against K562 and U937 cell-lines was increased by both GM-CSF and IL-3 stimulation, and both cytokines primed monocytes for increased killing of K562 and KG-1 cell-lines as well as primary AML blasts in response to LPS. The mechanism of action of MMC was largely confirmed to be via surface-bound TNF, although other TNF-independent mechanisms must have been involved.


Subject(s)
Cytotoxicity, Immunologic/drug effects , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Interleukin-3/pharmacology , Leukemia/immunology , Monocytes/immunology , Tumor Necrosis Factor-alpha/biosynthesis , Humans , L-Lactate Dehydrogenase/metabolism , Monocytes/drug effects , Monocytes/metabolism , Receptors, Tumor Necrosis Factor/biosynthesis
18.
Bone Marrow Transplant ; 26(11): 1165-72, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11149726

ABSTRACT

Between October 1993 and March 1999, 29 patients with CML who were ineligible for allogeneic BMT underwent PBSC harvest using idarubicin, cytarabine and G-CSF. In 9/29 (31%) patients all collected stem cells were Ph-negative, and 15/29 patients' (52%) collections were substantially (>95%) Ph-negative. The proportion of patients from whom Ph-negative stem cells were obtained was similar between patients who had, or had not, received prior alphaIFN. Fifteen patients in chronic phase (median age 45) proceeded to PBSCT following busulphan 16 mg/m2 and cyclophosphamide 120 mg/m2. Nine of the 13 patients who had failed to respond to prior alphaIFN proceeded to stem cell transplantation as soon as was feasible and six of the newly diagnosed patients were transplanted after failing to achieve a cytogenetic response after a minimum of 12 months on alphaIFN following progenitor cell harvest. The median number of days to neutrophils >0.5 and platelet >50 was 18 (range 13-69) and 28 (range 13-234), respectively. There was no procedure-related mortality. At median follow-up of 2.3 years post autograft 10 of 15 patients remain alive and in chronic phase. Overall survival for all 27 patients at 5 years after initial diagnosis is 70% and median survival from diagnosis 7.3 years. Survival for alphaIFN non-responders who were transplanted is 74% at 5 years from diagnosis and 75% at 3 years from transplant. Cytogenetic analysis performed 3 months post transplant demonstrated one patient with a complete cytogenetic response, seven with a partial response and three with no response. Six patients remain partially Ph-negative, with one major CR. Survival for all patients in the protocol is favourable compared with conventional therapy and is particularly encouraging following PBSCT for alphaIFN non-responsive patients. Patients not responding to alphaIFN can be induced into Ph-negativity with PBSCT but this may not always be sustainable. There seems to be no obvious disadvantage in harvesting stem cells after prior exposure to alphaIFN, providing an adequate alphaIFN-free rest period is used.


Subject(s)
Hematopoietic Stem Cell Transplantation , Interferon-alpha/therapeutic use , Leukemia, Myeloid, Chronic-Phase/therapy , Adult , Antineoplastic Combined Chemotherapy Protocols/adverse effects , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Busulfan/administration & dosage , Combined Modality Therapy , Cyclophosphamide/administration & dosage , Cytarabine/administration & dosage , Female , Granulocyte Colony-Stimulating Factor/administration & dosage , Hematopoietic Stem Cell Mobilization , Hematopoietic Stem Cell Transplantation/adverse effects , Hematopoietic Stem Cells/ultrastructure , Humans , Idarubicin/administration & dosage , Leukemia, Myeloid, Chronic-Phase/drug therapy , Leukemia, Myeloid, Chronic-Phase/genetics , Male , Middle Aged , Philadelphia Chromosome , Prospective Studies , Survival Rate , Treatment Outcome
19.
Hematology ; 5(1): 27-39, 2000.
Article in English | MEDLINE | ID: mdl-11399599

ABSTRACT

Myeloma accounts for 1% of all malignancies and is characterised by the malignant expansion of plasma cells. It is essentially incurable with a median survival of 6 months untreated. Typically disseminated at presentation, myeloma requires systemic therapy, in those patients with advanced or progressive disease. Of single agent therapies, oral mephalan remains the gold standard, either alone or in combination with steroids. 40% will respond with an increase in median survival from under 1 year to around 36 months. Combination chemotherapy produces an advantage in remission induction but no clear benefit for either survival or remission duration has been shown over standard melphalan and prednisolone. Significant melphalan dose escalation produces increased response rates and increased complete remission rates, however at the expense of increased toxicity. This has been ameliorated to an extent using stem cell support, however, although autografting may be superior to conventional chemotherapy it is not a curative treatment and modifications such as purging or tandem procedures remain to be fully evaluated. Allogeneic transplantation is only an option in a tiny minority of patients and has a high treatment-related mortality. Interferon alpha may improve the relapse free survival but has little effect on overall survival as the majority of patients will relapse. Experimental approaches, such as P-gp inhibition or thalidomide, are appropriate in this poor prognosis group. In many patients palliative care remains the only therapeutic option and close attention should be therefore paid to the prevention and treatment of renal failure, anaemia, infection, hypercalcaemia and bone disease.

20.
Br J Haematol ; 107(4): 861-9, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10606895

ABSTRACT

Cells of solid tumours tend to rely on glycolysis for energy. On the other hand, increased glycolysis in solid tumour cells expressing the multidrug resistance protein MDR-1 has been associated with increased malignancy in tumours. We have previously shown that cells of the MDR-1-positive CEM/VLB100 leukaemic cell line have increased mitochondrial electron transport chain (mtETC) activity compared with parental CEM cells. In the present study we used infrared (IR) spectroscopy to demonstrate that the mitochondrial DNA (mtDNA) content in the CEM/VLB100 cell line was significantly increased compared to that in the parental CEM cells. The increase in mtDNA was not accompanied by an increase in mitochondrial protein as both lipid and protein levels were decreased in CEM/VLB100 mitochondria. The ATP content was similar in these two cell lines. However, the ATP-dependent membrane efflux pump function in CEM/VLB100 cells was significantly reduced when mitochondrial ATP synthesis was inhibited by oligomycin, a specific inhibitor of mitochondrial F0F1-ATPase. Proliferation of CEM/VLB100 cells was significantly decreased compared to parental CEM cells, and was independent of p53 expression. Thus, we conclude that: (1) IR spectroscopy is a potential powerful technique for detecting mtDNA, protein and lipid contents simultaneously; (2) leukaemic cells mainly rely on mtDNA for energy; (3) increased expression of an ATP-dependent membrane efflux pump such as Pgp may up-regulate ATP generation and mtDNA content. These metabolic perturbations may exist merely to serve the efflux pump and do not result in an increase in leukaemic cell proliferation. In addition, the associated reduction in mitochondrial lipid and protein may contribute to sensitize the cells to cytochrome c release.


Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , DNA, Mitochondrial/metabolism , Leukemia/metabolism , Cell Division/physiology , Drug Resistance, Multiple , Humans , Leukemia/pathology , Tumor Cells, Cultured
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