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Title: Fluorescence in situ hybridization (FISH) study of response to granulocyte colony stimulating factor (G-CSF) in myelodysplasia associated with monosomy 7: Evidence for differentiation of the dysplastic clone

Abstract

Myelodysplastic syndromes (MDS) are clonal disorders of hematopoietic differentiation which may respond to the administration of cytokines with increases in the number of circulating mature neutrophils. We used fluorescence in situ hybridization (FISH) to investigate a young patient with myelodyplasia associated with monosomy 7 to determine whether mature appearing polymorphonuclear cells present in response to treatment with granulocyte colony stimulating factor (G-CSF) were progeny of the dysplastic clone or represented stimulation of residual normal hematopoiesis. The patient is a 26 year old male with a long history of a complex stem cell disorder dating to age 5. A chromosome fragility test was negative. In October 1993 treatment was begun with G-CSF when the absolute neutrophil count (ANC) fell to 300/mm{sup 3} despite GM-CSF therapy. Cytogenetic study of bone marrow just prior to starting G-CSF revealed monosomy 7 in all metaphases. A study in July, 1992 was normal. On his most recent marrow examination (1/94), blasts had numerous dysplastic forms. FISH was performed on buffy coat smears of patient and control specimens using a biotin labelled alpha satellite probe to chromosome 7. At the time of study, the peripheral blood count was 12,500 mm{sup 3}, with 56% neutrophils, 6% bands andmore » no circulating blasts. Cells were scored as either polymorphonuclear or mononuclear cells. In a healthy control, 22 of 190 scored as either polymorphonuclear cells (12%) contained one chromosome 7 signal, versus 193 of 200 (96.5%) in the patient. For mononuclear cells, the control demonstrated 23 of 137 nuclei (17%) with one signal, versus 300 of 511 nuclei (59%) in the patient. We conclude that G-CSF induced differentiation in the dysplastic clone in this case and did not stimulate normal hermatopoiesis.« less

Authors:
 [1]; ;  [2]
  1. Rhode Island Hospital, Providence, RI (United States)
  2. Brown Univ. School of Medicine, Providence, RI (United States)
Publication Date:
OSTI Identifier:
133484
Report Number(s):
CONF-941009-
Journal ID: AJHGAG; ISSN 0002-9297; TRN: 95:005313-0212
Resource Type:
Journal Article
Resource Relation:
Journal Name: American Journal of Human Genetics; Journal Volume: 55; Journal Issue: Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; PATIENTS; BLOOD FORMATION; HEMIC DISEASES; HUMAN CHROMOSOME 7; DNA-CLONING; CHROMOSOMAL ABERRATIONS; DNA HYBRIDIZATION; FLUORESCENCE; HEMATOPOIETIC SYSTEM; LEUKOCYTES; PROBES

Citation Formats

Cicilline, M., Mark, H.F.L., and Rintels, P. Fluorescence in situ hybridization (FISH) study of response to granulocyte colony stimulating factor (G-CSF) in myelodysplasia associated with monosomy 7: Evidence for differentiation of the dysplastic clone. United States: N. p., 1994. Web.
Cicilline, M., Mark, H.F.L., & Rintels, P. Fluorescence in situ hybridization (FISH) study of response to granulocyte colony stimulating factor (G-CSF) in myelodysplasia associated with monosomy 7: Evidence for differentiation of the dysplastic clone. United States.
Cicilline, M., Mark, H.F.L., and Rintels, P. 1994. "Fluorescence in situ hybridization (FISH) study of response to granulocyte colony stimulating factor (G-CSF) in myelodysplasia associated with monosomy 7: Evidence for differentiation of the dysplastic clone". United States. doi:.
@article{osti_133484,
title = {Fluorescence in situ hybridization (FISH) study of response to granulocyte colony stimulating factor (G-CSF) in myelodysplasia associated with monosomy 7: Evidence for differentiation of the dysplastic clone},
author = {Cicilline, M. and Mark, H.F.L. and Rintels, P.},
abstractNote = {Myelodysplastic syndromes (MDS) are clonal disorders of hematopoietic differentiation which may respond to the administration of cytokines with increases in the number of circulating mature neutrophils. We used fluorescence in situ hybridization (FISH) to investigate a young patient with myelodyplasia associated with monosomy 7 to determine whether mature appearing polymorphonuclear cells present in response to treatment with granulocyte colony stimulating factor (G-CSF) were progeny of the dysplastic clone or represented stimulation of residual normal hematopoiesis. The patient is a 26 year old male with a long history of a complex stem cell disorder dating to age 5. A chromosome fragility test was negative. In October 1993 treatment was begun with G-CSF when the absolute neutrophil count (ANC) fell to 300/mm{sup 3} despite GM-CSF therapy. Cytogenetic study of bone marrow just prior to starting G-CSF revealed monosomy 7 in all metaphases. A study in July, 1992 was normal. On his most recent marrow examination (1/94), blasts had numerous dysplastic forms. FISH was performed on buffy coat smears of patient and control specimens using a biotin labelled alpha satellite probe to chromosome 7. At the time of study, the peripheral blood count was 12,500 mm{sup 3}, with 56% neutrophils, 6% bands and no circulating blasts. Cells were scored as either polymorphonuclear or mononuclear cells. In a healthy control, 22 of 190 scored as either polymorphonuclear cells (12%) contained one chromosome 7 signal, versus 193 of 200 (96.5%) in the patient. For mononuclear cells, the control demonstrated 23 of 137 nuclei (17%) with one signal, versus 300 of 511 nuclei (59%) in the patient. We conclude that G-CSF induced differentiation in the dysplastic clone in this case and did not stimulate normal hermatopoiesis.},
doi = {},
journal = {American Journal of Human Genetics},
number = Suppl.3,
volume = 55,
place = {United States},
year = 1994,
month = 9
}
  • Supernatants of COS-1 cells transfected with gibbon cDNA encoding interleukin 3 (IL-3) with homology to sequences for human IL-3 were tested for ability to promote growth of various human hemopoietic progenitors. The effect of these supernatants as a source of recombinant IL-3 was compared to that of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) as well as to that of medium conditioned by phytohemagglutinin-stimulated leukocytes. The frequency of multilineage colonies, erythroid bursts, and megakaryocyte colonies in cultures containing the COS-1 cell supernatant was equivalent to the frequency observed in the controls and significantly higher than foundmore » in cultures plated with recombinant GM-CSF. G-CSF did not support the formation of multilineage colonies, erythroid bursts, and megakaryocyte colonies. In contrast, growth of granulocyte-macrophage colonies was best supported with GM-CSF, while recombinant IL-3 yielded colonies at lower or at best equivalent frequency. The simultaneous addition of higher concentrations of GM-CSF to cultures containing IL-3 in optimal amounts did not enhance the formation of multilineage colonies, erythroid bursts, and megakaryocyte colonies. However, the frequency of such colonies and bursts increased with GM-CSF when cultures were plated with suboptimal concentrations of IL-3. Growth of colonies within the granulocyte-macrophage lineage is optimally supported by GM-CSF and does not increase with further addition of IL-3.« less
  • Interleukin 1 alpha (IL-1 alpha), tumor necrosis factor alpha (TNF alpha), granulocyte-colony-stimulating factor (G-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) are molecularly distinct cytokines acting on separate receptors. The release of these cytokines can be concomitantly induced by the same signal and from the same cellular source, suggesting that they may cooperate. Administered alone, human recombinant (hr)IL-1 alpha and hrTNF alpha protect lethally irradiated mice from death, whereas murine recombinant GM-CSF and hrG-CSF do not confer similar protection. On a dose basis, IL-1 alpha is a more efficient radioprotector than TNF alpha. At optimal doses, IL-1 alpha is a more radioprotectivemore » cytokine than TNF alpha in C57BL/6 and B6D2F1 mice and less effective than TNF alpha in C3H/HeN mice, suggesting that the relative effectiveness of TNF alpha and IL-1 alpha depends on the genetic makeup of the host. Administration of the two cytokines in combination results in additive radioprotection in all three strains. This suggests that the two cytokines act through different radioprotective pathways and argues against their apparent redundancy. Suboptimal, nonradioprotective doses of IL-1 alpha also synergize with GM-CSF or G-CSF to confer optimal radioprotection, suggesting that such an interaction may be necessary for radioprotection of hemopoietic progenitor cells.« less
  • We have attempted to evaluate in vivo effects of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) on acute radiation hematopoietic injury in mice. BDF1 mice, irradiated with 7.5-Gy x-rays, were injected i.p. twice daily for 10 days with 10(5) U recombinant human G-CSF, 3.75 x 10(5) U recombinant murine GM-CSF, or a combination of both. G-CSF significantly enhanced the recovery of not only peripheral leukocytes but also platelets and hematocrit on days 14 and 21 after irradiation. GM-CSF significantly enhanced the recovery of platelets on day 14 and peripheral leukocytes on day 21. G-CSF markedly enhanced the recoverymore » of spleen colony-forming units (CFU-S), colony-forming units in culture (CFU-C), erythroid burst-forming units (BFU-E), and megakaryocyte colony-forming units (CFU-Meg) both in bone marrow and in the spleen. GM-CSF significantly enhanced the recovery of CFU-Meg in bone marrow on day 14. We found synergistic effects between G-CSF and GM-CSF on CFU-S, CFU-C, and CFU-Meg in the spleen on day 14, although we found antagonistic effects between G-CSF and GM-CSF on CFU-S and CFU-C in bone marrow on day 7, and on platelet counts on day 7. These results indicate that the administration of recombinant G-CSF and GM-CSF may be useful in accelerating hematopoietic recovery in patients with acute radiation hematopoietic injuries.« less
  • Purpose: Based on early clinical evidence of potential mucosal protection by granulocyte-macrophage colony stimulating factor (GM-CSF), the Radiation Therapy Oncology Group conducted a double-blind, placebo-controlled, randomized study to test the efficacy and safety of GM-CSF in reducing the severity and duration of mucosal injury and pain (mucositis) associated with curative radiotherapy (RT) in head-and-neck cancer patients. Methods and Materials: Eligible patients included those with head-and-neck cancer with radiation ports encompassing >50% of oral cavity and/or oropharynx. Standard RT ports were used to cover the primary tumor and regional lymphatics at risk in standard fractionation to 60-70 Gy. Concurrent cisplatin chemotherapymore » was allowed. Patients were randomized to receive subcutaneous injection of GM-CSF 250 {mu}g/m{sup 2} or placebo 3 times a week. Mucosal reaction was assessed during the course of RT using the National Cancer Institute Common Toxicity Criteria and the protocol-specific scoring system. Results: Between October 2000 and September 2002, 130 patients from 36 institutions were accrued. Nine patients (7%) were excluded from the analysis, 3 as a result of drug unavailability. More than 80% of the patients participated in the quality-of-life endpoint of this study. The GM-CSF did not cause any increase in toxicity compared with placebo. There was no statistically significant difference in the average mean mucositis score in the GM-CSF and placebo arms by a t test (p = 0.4006). Conclusion: This placebo-controlled, randomized study demonstrated no significant effect of GM-CSF given concurrently compared with placebo in reducing the severity or duration of RT-induced mucositis in patients undergoing definitive RT for head-and-neck cancer.« less
  • Physical mapping of small genomic DNA fragments or expressed sequences by in situ hybridization is typically limited by the size of the target DNA sequence. Isolation of large insert DNA clones from libraries containing the target DNA sequence facilitates physical mapping by fluorescence in situ hybridization and allows rapid assignment of genes to cytogenetic bands. Here, we demonstrate the scheme by mapping the human protooncogene trk (NTRK1), a tyrosine kinase receptor type I gene that has earlier been assigned to two different cytogenetic loci. Large DNA insert library screening was carried out by in vitro DNA amplification using oligonucleotide primersmore » flanking exon 4 of trk. The scheme presented here can easily be generalized to map physically very small nonrepetitive genomic DNA fragments or incomplete cDNAs. 17 refs., 2 figs.« less