Tissue-based assay for ornithine decarboxylase to identify patients likely to respond to difluoromethylornithine

Victor A. Levin, Jacob L. Jochec, Lisa M. Shantz, Patricia E. Koch, Anthony E. Pegg

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

In a previous publication, we showed that a clinical trial of DL-α-difluoromethyl ornithine (DFMO), in combination with PCV (procarbazine, CCNU, vincristine) increased survival of patients with anaplastic gliomas (WHO III) but not glioblastoma multiforme (WHO IV). We believe that treatment outcome (survival) is inversely related to tumor ornithine decarboxylase (ODC) levels. To prove this, we needed to develop an assay to quantify ODC levels in formalin-fixed tumor tissues, which would enable a retrospective study of tumor biopsy specimens from the landmark clinical trial. We developed an assay using a specific polyclonal antibody coupled to an Alexa fluorescent dye. Transgenic MHC-ODC mice with differing levels of ODC in heart muscle were used to establish the relationship between mean gray-scale intensity and enzymatic ODC activity. We found a direct relationship between mean gray-scale intensity of the ODC antibody coupled to Alexa 647 dye and enzymatic activity. Preliminary analysis of a human glioma tissue array shows that tumor-specific variations in levels of ODC can be semiquantitated. We show that mean gray-scale intensity of astrocytoma:glioblastoma is 1:6 and of anaplastic astrocytoma:glioblastoma is 1:4. We also compared the intensity of antibody to ki67 coupled with phycoerythrin simultaneously in cells but failed to see a relationship that crossed histologies. We conclude that we can measure levels of ODC in formalin-fixed tumor tissue using an antibody to ODC coupled to Alexa 647 dye, and this will enable us to conduct a future study to correlate survival of patients with gliomas of different histologies treated with DFMO to tumor ODC levels.

Original languageEnglish (US)
Pages (from-to)1467-1474
Number of pages8
JournalJournal of Histochemistry and Cytochemistry
Volume52
Issue number11
DOIs
StatePublished - Nov 1 2004

Fingerprint

Eflornithine
Ornithine Decarboxylase
Glioblastoma
Glioma
Neoplasms
Ornithine
Antibodies
Astrocytoma
Formaldehyde
Survival
Histology
Coloring Agents
Clinical Trials
Phycoerythrin
Lomustine
Procarbazine
Vincristine
Fluorescent Dyes
Myocardium
Retrospective Studies

All Science Journal Classification (ASJC) codes

  • Anatomy
  • Histology

Cite this

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title = "Tissue-based assay for ornithine decarboxylase to identify patients likely to respond to difluoromethylornithine",
abstract = "In a previous publication, we showed that a clinical trial of DL-α-difluoromethyl ornithine (DFMO), in combination with PCV (procarbazine, CCNU, vincristine) increased survival of patients with anaplastic gliomas (WHO III) but not glioblastoma multiforme (WHO IV). We believe that treatment outcome (survival) is inversely related to tumor ornithine decarboxylase (ODC) levels. To prove this, we needed to develop an assay to quantify ODC levels in formalin-fixed tumor tissues, which would enable a retrospective study of tumor biopsy specimens from the landmark clinical trial. We developed an assay using a specific polyclonal antibody coupled to an Alexa fluorescent dye. Transgenic MHC-ODC mice with differing levels of ODC in heart muscle were used to establish the relationship between mean gray-scale intensity and enzymatic ODC activity. We found a direct relationship between mean gray-scale intensity of the ODC antibody coupled to Alexa 647 dye and enzymatic activity. Preliminary analysis of a human glioma tissue array shows that tumor-specific variations in levels of ODC can be semiquantitated. We show that mean gray-scale intensity of astrocytoma:glioblastoma is 1:6 and of anaplastic astrocytoma:glioblastoma is 1:4. We also compared the intensity of antibody to ki67 coupled with phycoerythrin simultaneously in cells but failed to see a relationship that crossed histologies. We conclude that we can measure levels of ODC in formalin-fixed tumor tissue using an antibody to ODC coupled to Alexa 647 dye, and this will enable us to conduct a future study to correlate survival of patients with gliomas of different histologies treated with DFMO to tumor ODC levels.",
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Tissue-based assay for ornithine decarboxylase to identify patients likely to respond to difluoromethylornithine. / Levin, Victor A.; Jochec, Jacob L.; Shantz, Lisa M.; Koch, Patricia E.; Pegg, Anthony E.

In: Journal of Histochemistry and Cytochemistry, Vol. 52, No. 11, 01.11.2004, p. 1467-1474.

Research output: Contribution to journalArticle

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AU - Levin, Victor A.

AU - Jochec, Jacob L.

AU - Shantz, Lisa M.

AU - Koch, Patricia E.

AU - Pegg, Anthony E.

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