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PMID: 16804528 Published · ppublish English Comparative Study Journal Article Validation Study

Method validation and preliminary qualification of pharmacodynamic biomarkers employed to evaluate the clinical efficacy of an antisense compound (AEG35156) targeted to the X-linked inhibitor of apoptosis protein XIAP.

British journal of cancer ·Vol. 95 ·No. 1 ·2006-07-03 ·Pages 42-8

Cummings J, Ranson M, Lacasse E, Ganganagari JR, St-Jean M, Jayson G, Durkin J, Dive C

Abstract

Data are presented on pharmacodynamic (PD) method validation and preliminary clinical qualification of three PD biomarker assays. M65 Elisa, which quantitates different forms of circulating cytokeratin 18 (CK18) as putative surrogate markers of both apoptotic and nonapoptotic tumour cell death, was shown to be highly reproducible: calibration curve linearity r2 = 0.996, mean accuracy > 91% and mean precision < 3%, n = 27. Employing recombinant (r) CK18 and caspase cleaved CK18 (CK18 Asp396 neo-epitope) as external standards, kit to kit reproducibly was < 6% (n = 19). rCK18 was stable in plasma for 4 months at -20 degrees C and -80 degrees C, for 4 weeks at 4 degrees C and had a half-life of 2.3 days at 37 degrees C. Cytokeratin 18 Asp396 NE, the M30 Apoptosense Elisa assay antigen, was stable in plasma for 6 months at -20 degrees C and -80 degrees C, for 3 months at 4 degrees C, while its half-life at 37 degrees C was 3.8 days. Within-day variations in endogenous plasma concentrations of the M30 and M65 antigens were assessed in two predose blood samples collected from a cohort of 15 ovarian cancer patients receiving carboplatin chemotherapy and were shown to be no greater than the variability associated with methods themselves. Between-day fluctuations in circulating levels of the M30 and M65 antigens and in XIAP mRNA levels measured in peripheral blood mononuclear cells by quantitative (q) RT-PCR were evaluated in two predose blood samples collected with a 5- to 7-day gap from 23 patients with advanced cancer enrolled in a phase I trial. The mean variation between the two pretreatment values ranged from 13 to 14 to 25%, respectively, for M65, M30 and qRT-PCR. These data suggest that the M30 and M65 Elisa's and qRT-PCR as PD biomarker assays have favourable performance characteristics for further investigation in clinical trials of anticancer agents which induce tumour apoptosis/necrosis or knockdown of the anti-apoptotic protein XIAP.

MeSH Terms
Apoptosis/drug effects Biomarkers, Tumor/blood Calibration Enzyme-Linked Immunosorbent Assay/methods Epitopes/blood,immunology Female Humans Keratin-18/blood,immunology Leukocytes, Mononuclear/chemistry,metabolism Oligonucleotides/pharmacokinetics Ovarian Neoplasms/blood,drug therapy Peptide Fragments/blood,immunology RNA, Messenger/antagonists & inhibitors,genetics,metabolism Reagent Kits, Diagnostic/standards Reference Values Reproducibility of Results Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity Time Factors X-Linked Inhibitor of Apoptosis Protein/antagonists & inhibitors,genetics
Chemicals
AEG 35156 Biomarkers, Tumor Epitopes Keratin-18 M65 antigen, human Oligonucleotides Peptide Fragments RNA, Messenger Reagent Kits, Diagnostic X-Linked Inhibitor of Apoptosis Protein XIAP protein, human
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Cummings J
Clinical and Experimental Pharmacology, Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester M20 4BX, England, UK. jcummings@picr.man.ac.uk
Ranson M
Lacasse E
Ganganagari J R
St-Jean M
Jayson G
Durkin J
Dive C
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Article Info
Journal
British journal of cancer
Abbr.
Br J Cancer
ISSN
0007-0920
DOI
10.1038/sj.bjc.6603220
Email
jcummings@picr.man.ac.uk
Published
2006-07-03
Pages
42-8
Language
English
Region
England
NLM ID
0370635
PMCID
PMC2360484
Subset
IM
Grants
Medical Research Council · G0601746 · United Kingdom
External Links
PubMed source DOI: 10.1038/sj.bjc.6603220 PMC Full Text View journal details
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