Inactivation of aldophosphamide by human aldehyde dehydrogenase isozyme 3

Francesco Giorgianni, Peter K. Bridson, Brian P. Sorrentino, Jörg Pohl, Raymond L. Blakley

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Tumors resistant to chemotherapeutic oxazaphosphorines such as cyclophosphamide often overexpress aldehyde dehydrogenase (ALDH), some isozymes of which catalyze the oxidization of aldophosphamide, an intermediate of cyclophosphamide activation, with formation of inert carboxyphosphamide. Since resistance to oxazaphosphorines can be produced in mammalian cells by transfecting them with the gene for human ALDH isozyme 3 (hALDH3), it seems possible that patients receiving therapy for solid tumors with cyclophosphamide might be protected from myelosuppression by their prior transplantation with autologous bone marrow that has been transduced with a retroviral vector causing overexpression of hALDH3. We investigated whether retroviral introduction of hALDH3 into a human leukemia cell line confers resistance to oxazaphosphorines. This was examined in the polyclonal transduced population, that is, without selecting out high expression clones. hALDH3 activity was 0.016 IU/mg protein in the transduced cells (compared with 2x 10-5 IU/mg in untransduced cells), but there was no detectable resistance to aldophosphamide-generating compounds (mafosfamide or 4- hydroperoxycyclophosphamide). The lack of protection was due, in part, to low catalytic activity of hALDH3 towards aldophosphamide, since, with NAD as cofactor, the catalytic efficiency of homogeneous, recombinant hALDH3 for aldophosphamide oxidation was shown to be about seven times lower than that of recombinant hALDH1. The two polymorphic forms of hALDH3 had identical kinetics with either benzaldehyde or aldophosphamide as substrate. Results of initial velocity measurements were consistent with an ordered sequential mechanism for ALDH1 but not for hALDH3; a kinetic mechanism for the latter is proposed, and the corresponding rate equation is presented. (C) 2000 Elsevier Science Inc.

Original languageEnglish (US)
Pages (from-to)325-338
Number of pages14
JournalBiochemical Pharmacology
Volume60
Issue number3
DOIs
StatePublished - Aug 1 2000
Externally publishedYes

Fingerprint

Aldehyde Dehydrogenase
Isoenzymes
Cyclophosphamide
perfosfamide
Tumors
Cells
aldophosphamide
human ADH1C protein
Kinetics
Autologous Transplantation
Human Activities
Velocity measurement
NAD
Catalyst activity
Neoplasms
Bone
Leukemia
Clone Cells
Genes
Bone Marrow

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Pharmacology

Cite this

Inactivation of aldophosphamide by human aldehyde dehydrogenase isozyme 3. / Giorgianni, Francesco; Bridson, Peter K.; Sorrentino, Brian P.; Pohl, Jörg; Blakley, Raymond L.

In: Biochemical Pharmacology, Vol. 60, No. 3, 01.08.2000, p. 325-338.

Research output: Contribution to journalArticle

Giorgianni, Francesco ; Bridson, Peter K. ; Sorrentino, Brian P. ; Pohl, Jörg ; Blakley, Raymond L. / Inactivation of aldophosphamide by human aldehyde dehydrogenase isozyme 3. In: Biochemical Pharmacology. 2000 ; Vol. 60, No. 3. pp. 325-338.
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