Your Logo Here CYTOCHROME P 450 3 A

Your Logo Here CYTOCHROME P 450 3 A AS A MEDIATOR OF CHEMOTHERAPY RESISTANCE IN PATIENTS WITH SOFT TISSUE SARCOMA

DISEASE-FREE SURVIVAL BY IFOSFAMIDE DOSE Worden, JCO (2005)

ERYTHROMYCIN BREATH TEST • [14 C N-methyl] erythromycin • CYP 3 A metabolizes erythromycin, liberating a carbon atom as CO 2 • Measures in vivo activity of CYP 3 A Dose 6 12 mean % range % 2. 4 0. 87 -4. 32 2. 2 0. 43 -4. 17

CYTOCHROME P 450 CYP 3 A Normal Liver Modified after Furlanut and Franceschi (2003) Oncology 65(Suppl 2): 2 -6. • Most abundant p 450 in liver • Composed of four family members – 3 A 4 – 3 A 5 – 3 A 7 – 3 A 43 • Responsible for metabolism of: – Ifosfamide – Vinblastine – Doxorubicin – Etoposide

ERMBT OVERALL SURVIVAL P value 0. 0992 P value 0. 0526

CYP 3 A QIF AND ERMBT r 2 5. 84 x 10 -6

QUANTITATIVE IMMUNOFLUORESCENCE (QIF) • Modification of Indirect IF method • Diminish auto fluorescence with glycine/PPD • Antibody labeled with Fluor 568 • Nuclei labeled with Syto 16 • Fluorescence analyzed by Mercury lamp with appropriate filter • Results expressed as F/nuclei

QIF RELIABILITY STUDIES INTRA-ASSAY VARIABILITY INTER-ASSAY VARIABILITY

BIOLOGICAL STUDIES • 60/79 patients with FFPE tissue available for study – TMA made • 19 patients with multiple specimens • 45/79 patients DNA extracted from FFPE tissue for SNP analysis

QIF CYP 3 A AND TUMOR HISTOLOGY

TUMOR CYP 3 A QIF AND SURVIVAL P value 0. 0353*

TUMOR CYP 3 A QIF AND SURVIVAL P value 0. 0415*

CONCLUSIONS • Decreased ERMBT are suggestive of poorer prognosis. • Elevated tumor CYP 3 A protein levels are associated with poor survival. • No correlations with CYP 3 A 4 or CYP 3 A 5 SNPs. • Prospectively studying patients for CYP 3 A levels in new clinical trial. • Examining the role of CYP 3 A gene induction in cell lines.

ACKNOWLEGMENTS • • • Larry BAKER Mark ZALUPSKI Tom GIORDANO Sybil BIERMANN Vernon SONDAK Rashmi CHUGH Walther Cancer Research Foundation Robert Urich Sarcoma Fund Staff and Patients at University of Michigan

Your Logo Here CYTOCHROME P 450 3 A AS A MEDIATOR OF CHEMOTHERAPY RESISTANCE IN PATIENTS WITH SOFT TISSUE SARCOMA