Critical issues in autologous bone marrow transplantation in adult acute leukemia.
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Adult↗
Biomedical subjects
M Beran
Publications and source records attributed to M Beran.
Regrowth of granulocyte-macrophage progenitor cells (GM-CFC) in suspension cultures of bone marrow depleted of GM-CFC with 4-hydroperoxycyclophosphamide (4-HC).
The hematopoietic capacity of bone marrow treated with 4-hydroperoxycyclophosphamide (4-HC) was studied using long-term suspension cultures. It was shown that, in the presence of a previously established marrow adherent cell layer (ACL), the 4-HC treated bone marrow, deprived of GM-CFC, generates GM-CFC in long-term suspension cultures in vitro. The kinetics of GM-CFC reappearance in in vitro cultures indicate their origin from more primitive progenitors rather than from surviving GM-CFC. In the presented modification, the suspension culture system is useful in evaluating the transplantation potential of bone marrow purged in vitro with chemotherapeutic or biologic agents.
Bone Marrow↗
Hematologic response of four patients with smoldering acute myelogenous leukemia to partially pure gamma interferon.
In vitro and in vivo studies were conducted to obtain basic information on the activity of gamma interferon (IFN-gamma) in acute myelogenous leukemia (AML). In a selected case of AML, recombinant IFN-gamma, but not IFN-alpha, induced differentiation of primary leukemic blasts in vitro. Similarly, IFN-gamma inhibited leukemic colony formation in vitro. This contrasted with IFN-alpha which was inactive. In one case of AML (M2), partially purified IFN-gamma given intravenously caused a shift of the WBC profile from immature blasts to maturing myeloid cells and neutrophil granulocytes. Intravenous IFN-gamma treatment of another patient who had AML as a second malignancy resulted in a complete hematologic remission, normalization of marrow granulocyte-macrophage colony-forming cell in vitro growth, and conversion of marrow cytogenetics from 95% hyperdiploid clone with complex abnormalities into 100% diploid. The results indicate a potential use of IFN-gamma in the treatment of selected patients with AML and the possibility of in vitro pretreatment evaluation of these patients' leukemic response to IFNs.
Adult↗
The localization of the human myeloperoxidase gene is in close proximity to the translocation breakpoint in acute promyelocytic leukemia.
The human myeloperoxidase (MPO) gene has recently been cloned in our laboratory. Southern blot hybridization of our MPO cDNA to DNA from a somatic cell hybrid clone panel revealed that the MPO cosegregated with human chromosome 17. In situ hybridization mapped the MPO gene to chromosome 17q22-24. Although this location is close to the translocation breakpoint which occurs in acute promyelocytic leukemia (APL), t(15;17)(q22;q21-22), Southern blot hybridization with different restriction-digested genomic DNA samples from four APL patients did not reveal MPO gene rearrangement. However, RNA dot-blot hybridization showed that APL patients with the translocation expressed high levels of MPO mRNA. This observation raises the possibility that the high levels of MPO gene expression in APL could be due to the arrest of leukemic cells at a specific stage of differentiation or a consequence of the translocation.
Chromosome Mapping↗
Diversity of the effect of recombinant tumor necrosis factors alpha and beta on human myelogenous leukemia cell lines.
In vitro characteristics of response to recombinant tumor necrosis factors alpha (rTNF-alpha) and beta (rTNF-beta) were studied in six human myelogenous leukemia cell lines. Heterogeneity of the response to rTNF was observed, and one line (K562) was resistant. No enhancement of cell growth was noted in any cell line. The dose-response curves for rTNF-alpha were characteristically sigmoid, the maximum inhibitory effect occurring between 25 and 200 ng/mL. Nonresponsiveness within this range indicated resistance that could not be overcome, even with very high doses of rTNF alpha. A similar response of sensitive and resistant lines occurred after exposure to rTNF-beta. The clonogenic cells were more sensitive than the overall population to the action of rTNF alpha, and prolonged exposure was necessary for manifestation of the effect. Concomitant exposure to recombinant interferon-alpha (rIFN-alpha) increased the response of two cell lines to rTNF-alpha, but no clear synergistic action could be demonstrated. The addition of rIFN-gamma was without effect. Variations in the rTNF-alpha-induced proliferative response could not be explained by differences in the number of binding sites per cell or their affinity for rTNF-alpha. That the clonogenic cells showed a higher sensitivity than the whole population might indicate a preferential effect on more primitive, actively proliferating cells with high growth potential.
Cell Division↗
The effect of long-term marrow culture on the nonadherent Philadelphia chromosome-positive clone in chronic myelogenous leukemia: preliminary observations.
To evaluate the effect of long-term culture on the Philadelphia chromosome-positive nonadherent hemopoietic clone in chronic myelogenous leukemia, bone marrow aspirates from five patients were grown in culture for three to four weeks. In addition to control cultures, samples were incubated with recombinant alpha-interferon. Cytogenetic studies performed on the nonadherent hemopoietic cells demonstrated suppression of the Philadelphia chromosome-positive clones, and appearance of cells with normal diploid karyotype in two of the five patients studied; cells from the remaining three patients showed persistent Philadelphia chromosome-positive cells in 100% of the metaphases. Exposure to alpha-interferon did not enhance the suppression of the Philadelphia chromosome-positive clones. A good correlation was noted between in vitro results and in vivo cytogenetic response: the two patients whose in vitro studies showed diploid clones achieved complete hematologic remission and suppression of Philadelphia chromosome-positive metaphases with therapy. Our data suggest that long-term culture favors the growth of normal clones over Philadelphia chromosome-positive clones in the nonadherent hemopoietic cells of some patients with chronic myelogenous leukemia. The value of the procedure as an in vitro predictive test for residual normal stem cells and response to therapy and for autologous marrow in vitro purging needs further exploration.
Adult↗
Clinical and prognostic features of Philadelphia chromosome-negative chronic myelogenous leukemia.
Between 1965 and 1982, 105 patients with a diagnosis of Philadelphia chromosome-negative chronic myelogenous leukemia were referred to our institution with minimal or no prior therapy. The median age was 63 years and 64% were males. The overall median survival from time of referral was 14 months; 53% of patients survived 1 year and only 10% survived beyond 5 years. At the time of analysis, 92 patients (88%) were dead, 56% of deaths being preceded by a blastic crisis. Compared with Philadelphia chromosome-positive disease, patients with Philadelphia chromosome-negative chronic myelogenous leukemia were older and had a significantly higher incidence of anemia, thrombocytopenia, monocytosis, marrow blasts, decreased marrow megakaryocytes and a lower incidence of basophilia and thrombocytosis. Chromosomal abnormalities occurred in 33% of patients and consisted most frequently of trisomy 8, or an additional chromosome C, loss of the Y chromosome, or abnormalities in chromosomes #5 and #7. Of nine pretreatment characteristics significantly associated with poor survival, a multivariate analysis identified four to have independent additive prognostic significance: severe thrombocytopenia, hemoglobin levels less than 10 g/dl, increasing peripheral blasts and promyelocytes, and age 60 years or older. Monocytosis was not of prognostic significance. The derived prognostic model divided patients into three risk groups, low, intermediate, and high, with median survivals of 36, 16, and 3 months, respectively. The authors conclude that Philadelphia chromosome-negative chronic myelogenous leukemia is a distinct entity among the myeloproliferative syndromes with characteristic clinical and laboratory features and a poor prognosis. Prognostic factors and related risk categories were demonstrated within this disease entity.
Adolescent↗
The association of specific "favorable" cytogenetic abnormalities with secondary leukemia.
Eight patients with secondary leukemia and specific cytogenetic abnormalities involving inversion of chromosome 16, or translocations between chromosomes 15 and 17, or 8 and 21, are presented. Seven of them (87%) achieved complete remission with chemotherapy. The occurrence of these karyotypes, which is unusual in secondary leukemia, and the favorable response to chemotherapy are discussed in relation to the pathogenesis of the disease.
Adult↗
The production of topoisomerase II-mediated DNA cleavage in human leukemia cells predicts their susceptibility to 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA).
Protein-associated DNA cleavage is produced in mammalian cells treated with active antileukemic DNA intercalating agents such as 4'(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA). We have examined the ability of m-AMSA to produce DNA cleavage in 3 human myeloid leukemic cell lines with different sensitivities to the cytotoxic actions of m-AMSA to see if the magnitude of DNA cleavage correlated with the degree of m-AMSA sensitivity. DNA alkaline elution was used to quantify DNA cleavage. The amount of m-AMSA-induced DNA cleavage in the two lines sensitive to m-AMSA was 1-2 orders of magnitude greater than that in an m-AMSA-resistant leukemic line. The m-AMSA resistant line had been developed by prolonged exposure of one of the sensitive lines to m-AMSA. This finding was not secondary to a decreased uptake of m-AMSA in the resistant cell line. m-AMSA treatment of the nuclei isolated from the three lines produced DNA cleavage frequencies comparable to the cleavage frequencies produced by m-AMSA treatment of the whole cells from which the nuclei were isolated. The DNA cleaving ability stimulated by m-AMSA is thought to be mediated by drug-induced effects on topoisomerase II, a nuclear enzyme that mediates alterations in DNA conformation. Alterations in the manner in which this enzyme interacts with antineoplastic agents may explain the emergence of resistant cells following initially successful chemotherapy.
Aminoacridines↗
The tissue localization of m-AMSA and its effect on thymidine incorporation in various tissues in vivo.
The distribution of 14C-labelled m-AMSA was studied in rats and pigmented mice using whole body autoradiography. The agent rapidly disappeared from the blood, accumulating in significant amounts in large parenchymal organs, certain endocrine tissues, and the retina of the pigmented mouse eye. The hemopoietic and lymphoid tissues showed a moderate uptake of radioactivity with the highest concentration observed in the thymus. The autoradiograms indicated a rapid excretion of radioactivity via the liver, kidney and the glandular part of the gastric mucosa. The distribution pattern of label from 14C-m-AMSA remained unaffected by pretreatment of animals with high dose (500 mg kg-1 b.w.) of cytosine arabinoside. Injection of unlabelled m-AMSA (7 mg kg-1 b.w.) to growing rats 24 h before sacrifice resulted in a highly significant (P less than 0.001) inhibition of 3H-thymidine incorporation into the DNA of thymus and spleen. A less pronounced reduction was observed in the kidney, adrenal, lung and testes. The thymidine incorporation into the DNA of bone marrow was markedly suppressed when calculated per dry weight, but increased when related to the DNA content, suggesting early regeneration of the remaining cells. In contrast, no significant effects were observed on the DNA synthesis in small intestine and liver.
Aminoacridines↗
The c-abl, bcr and C lambda genes are amplified in a cell line but not in the uncultured cells from a patient with chronic myelogenous leukemia.
Structural alterations of the oncogenes in human tumors are reported to result from a variety of mechanisms: point mutations, chromosomal translocations and gene amplifications. In over 90% of the cases of chronic myelogenous leukemia (CML), the c-abl oncogene is translocated from chromosome 9 to chromosome 22, and forms in part the Philadelphia (Ph1) chromosome. We have molecularly analysed a double Ph1-positive (Ph1+) cell line, KBM-5 that was established from a patient with CML in the blast-transformed phase (CML-BP). We report that the c-abl, bcr, and C lambda genes are amplified approximately eight-fold in the cell line but not in the fresh uncultured cells from which KBM-5 was derived.
Cell Line↗
Induction of differentiation in human myeloid leukemia cells with cytosine arabinoside.
Normalization of granulocyte counts was obtained with low-dose cytosine arabinoside (Ara-C) in a patient with acute myelogenous leukemia (AML, FAB-M2) who had relapsed on maintenance therapy containing conventional doses of Ara-C and was clinically resistant to high-dose Ara-C treatment. The patient's leukemic cells exhibited an abnormal karyotype (45, X-Y, t[8q; 21q]). To determine whether the response to low-dose Ara-C was a result of induced leukemic cell differentiation, the mature peripheral blood granulocytes were isolated and fused with mitotic chinese hamster ovary (CHO) cells to induce premature chromosome condensation. Karyotypic evaluation of the granulocyte prematurely condensed chromosomes (PCC) during response to low-dose Ara-C demonstrated the presence of cells with 45 chromosomes. This result strongly suggests that part of this patient's response to low-dose Ara-C was a result of induced maturation of the leukemic clone.
Bone Marrow Cells↗
Analysis of nuclear m-AMSA content by DNA fluorochrome competition.
Spectrofluorometry and flow cytometry were used to measure cellular uptake of the anti-leukemic drug 4'-(9-acridinylamino) methane sulfon-m-anisidide (m-AMSA). Because of its very low intrinsic fluorescence, we used m-AMSA to quench DNA-fluorescence induced by the vital DNA fluorochrome Hoechst 33342. Maximum fluorescence was obtained with cells incubated in the fluorochrome alone. Subsequent incubation of cells in increasing concentrations of m-AMSA resulted in a gradual decrease in fluorescence. Upon incubation in drug-free medium, the quenching phenomenon was reversible, consistent with rapid exit of m-AMSA from the cells. The novel competitive fluorescence assay for cellular uptake for m-AMSA showed a better correlation to nuclear accumulation of the drug, than to its overall cellular accumulation, which may be important in assessment of cellular resistance to m-AMSA, with possible low nuclear accumulation of the drug. When this competitive fluorescence technique for measurement of cellular m-AMSA concentration was applied in flow-cytometric setting, subpopulations of normal human white blood cells were detected with distinctly different fluorescence patterns, indicating differences in cellular m-AMSA uptake. The potential use of this technique is to detect differences between cell subpopulations with different drug uptake abilities.
Amsacrine↗
Altered expression of G1-specific genes in human malignant myeloid cells.
We have studied the expression of cell-cycle genes specific to the G1 (2A9, 2F1, 4F1, c-myc) and S (histone H3) phases of the cell cycle in normal and malignant human myeloid cycling cells. The levels of expression were determined by measuring the amounts of specific RNA in blot hybridization assays. Levels of expression of the G1 genes were compared to the level of expression of the S-phase-specific H3 gene. This method can distinguish whether an increased expression of G1 genes is truly due to deregulation or simply reflects an increase in the fraction of proliferating cells. In a normal asynchronous system provided by the bone marrow cells of three normal donors, the expressions of the four G1-specific genes 2A9, 2F1, 4F1, and c-myc, and of the S-phase-specific gene H3 were in ratios that differed little from one individual to another. In the total RNA of eight patients in the chronic phase of chronic myelogenous leukemia, a high level of expression of G1 cell-cycle genes was paralleled by a high level of expression of the S-phase H3 gene, simply reflecting an increase in the fraction of proliferating cells. In patients with acute myelogenous leukemia (AML), the RNA levels of 2F1 and 4F1 paralleled the expression of H3-i.e., the ratios of expression 2F1/H3 and 4F1/H3 were the same as in normal bone marrow cells. However, in 9 of 10 patients with AML we found that the expression of c-myc was elevated with respect to H3 expression. The expression of 2A9 (with respect to H3) was also elevated in some of these AML patients. Two important conclusions can be drawn from these findings: increased levels of a G1-specific RNA in a tumor may not indicate overexpression of that gene but may instead simply reflect the fraction of proliferating cells; and in some patients with AML, however, the expression of certain G1 genes is truly deregulated and might contribute to the impairment of proliferative control that is associated with this phenotype.
Cell Cycle↗
Transection of varicose vessels in the nasal mucosa of patients with recurrent epistaxis. A 2-year follow-up.
Twenty-two patients with recurrent nose bleeds and varicose vessels on the nasal septum were subjected to transection of the vessels and followed-up for 2 years. One month after the operation, it was not possible to identify the original vessels. In other locations new varicose vessels began to appear after 1 month, and at the end of the follow-up period were observed on 9 out of 10 operated sides. The transection resulted in an immediate decrease in the frequency of nose bleeds and at the end of the control period 12 (55%) of the patients had had less than 4 bleeds during the previous year in spite of the observed rapid formation of new vessels. Mucosal atrophy with symptoms of increased nasal crusting occurred in 3 patients. Transection of varicose vessels in patients with nose bleeds leads to a decrease in the number of bleeds.
Adult↗
Occurrence of epistaxis in habitual nose-bleeders and analysis of some etiological factors.
The occurrence of epistaxis in 121 habitual nose-bleeders has been analysed together with the importance of different etiological factors. Heredity for recurrent nose-bleeds was noted by 42% of the bleeders, which was significantly more than in the control group (p less than 0.001). The start of the nose-bleeds was mostly spontaneous. Both common cold and stress or tiredness were frequently experienced before the occurrence of the nose-bleeds. The blood pressure distribution of the habitual nose-bleeders did not differ from that of the population samples used for comparison. Diseases reported by the habitual bleeders were few, and the blood routine tests revealed only few diseases that could be related to the recurrent nose-bleeds.
Adolescent↗
Leukemia and neutrophilic dermatosis: report of a case.
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Adult↗
Drug sensitivity and cross-resistance of the 4'-(9-acridinylamino)methanesulfon-m-anisidide-resistant subline of HL-60 human leukemia.
A subline of the HL-60 leukemia resistant to 4'-(9-acridinylamino)methanesulfon-m-anisidide (HL-60/AMSA) was developed by intermittent long-term in vitro treatment. Resistance to 4'-(9-acridinylamino)methanesulfon-m-anisidide remained unchanged after 180 doublings in the absence of the drug, suggesting a stable phenotypic alteration. The pattern of cross-resistance of HL-60/AMSA was evaluated for a spectrum of antileukemic agents using the clonogenic assay. Modest cross-resistance to doxorubicin (Adriamycin) was observed in the resistant subline on continuous exposure to the drug for 8 to 9 days; however, HL-60/AMSA cells retained their sensitivity to doxorubicin following short-term exposure for 60 min. HL-60/AMSA was also sensitive to the anthracycline aclacinomycin, Vinca alkaloids, and alkylating agents. Furthermore, enhanced sensitivity to 1-beta-D-arabinofuranosylcytosine was observed. The subline was cross-resistant to etoposide.
Aminoacridines↗