A Novel Biomarker Panel Examining Response to Gemcitabine with or without Erlotinib for Pancreatic Cancer Therapy in NCIC Clinical Trials Group PA.3

David B Shultz, Jonathan Pai, Wayland Chiu, Kendall Ng, Madeline G Hellendag, Gregory Heestand, Daniel T Chang, Dongsheng Tu, Malcolm J Moore, Wendy R Parulekar, Albert C Koong, David B Shultz, Jonathan Pai, Wayland Chiu, Kendall Ng, Madeline G Hellendag, Gregory Heestand, Daniel T Chang, Dongsheng Tu, Malcolm J Moore, Wendy R Parulekar, Albert C Koong

Abstract

Purpose: NCIC Clinical Trials Group PA.3 was a randomized control trial that demonstrated improved overall survival (OS) in patients receiving erlotinib in addition to gemcitabine for locally advanced or metastatic pancreatic cancer. Prior to therapy, patients had plasma samples drawn for future study. We sought to identify biomarkers within these samples.

Experimental design: Using the proximity ligation assay (PLA), a probe panel was built from commercially available antibodies for 35 key proteins selected from a global genetic analysis of pancreatic cancers, and used to quantify protein levels in 20 uL of patient plasma. To determine if any of these proteins levels independently associated with OS, univariate and mulitbaraible Cox models were used. In addition, we examined the associations between biomarker expression and disease stage at diagnosis using Fisher's exact test. The correlation between Erlotinib sensitivity and each biomarkers was assessed using a test of interaction between treatment and biomarker.

Results and conclusion: Of the 569 eligible patients, 480 had samples available for study. Samples were randomly allocated into training (251) and validation sets (229). Among all patients, elevated levels of interleukin-8 (IL-8), carcinoembryonic antigen (CEA), hypoxia-inducible factor 1-alpha (HIF-1 alpha), and interleukin-6 were independently associated with lower OS, while IL-8, CEA, platelet-derived growth factor receptor alpha and mucin-1 were associated with metastatic disease. Patients with elevated levels of receptor tyrosine-protein kinase erbB-2 (HER2) expression had improved OS when treated with erlotinib compared to placebo. In conclusion, PLA is a powerful tool for identifying biomarkers from archived, small volume serum samples. These data may be useful to stratify patient outcomes regardless of therapeutic intervention.

Trial registration: ClinicalTrials.gov NCT00040183.

Conflict of interest statement

Competing Interests: The authors have read the journal's policy and the authors of this manuscript have the following competing interests: GH has served as a consultant for Bayer, Merrimack, and Genomic Health. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1. Univariate (A) and multivariable (B)…
Fig 1. Univariate (A) and multivariable (B) analyses of the association between biomarker concentration, according to median levels, and overall survival.
Hazard ratios (HR) (of high to low biomarker concentration) and p-values are shown for biomarkers that were significantly associated (p

Fig 2. Odds ratios (OR) of having…

Fig 2. Odds ratios (OR) of having stage III disease (locally advanced) for biomarker concentration,…

Fig 2. Odds ratios (OR) of having stage III disease (locally advanced) for biomarker concentration, according to median levels (low vs. high).
ORs and p-values are shown for biomarkers that were significantly associated (p

Fig 3. Erlotinib sensitivity is associated with…

Fig 3. Erlotinib sensitivity is associated with Her2 expression.

Median overall survival (with 95% CI)…

Fig 3. Erlotinib sensitivity is associated with Her2 expression.
Median overall survival (with 95% CI) was improved with erlotnib treatment in patients with higher than median levels of plasma HER2 (8.2 vs. 5.0 months) but not in patients with lower plasma HER2 (6.0 vs. 8.3 months).
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References
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Funding for this study was provided by My Blue Dots and the Sydney Ng Memorial Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Fig 2. Odds ratios (OR) of having…
Fig 2. Odds ratios (OR) of having stage III disease (locally advanced) for biomarker concentration, according to median levels (low vs. high).
ORs and p-values are shown for biomarkers that were significantly associated (p

Fig 3. Erlotinib sensitivity is associated with…

Fig 3. Erlotinib sensitivity is associated with Her2 expression.

Median overall survival (with 95% CI)…

Fig 3. Erlotinib sensitivity is associated with Her2 expression.
Median overall survival (with 95% CI) was improved with erlotnib treatment in patients with higher than median levels of plasma HER2 (8.2 vs. 5.0 months) but not in patients with lower plasma HER2 (6.0 vs. 8.3 months).
Fig 3. Erlotinib sensitivity is associated with…
Fig 3. Erlotinib sensitivity is associated with Her2 expression.
Median overall survival (with 95% CI) was improved with erlotnib treatment in patients with higher than median levels of plasma HER2 (8.2 vs. 5.0 months) but not in patients with lower plasma HER2 (6.0 vs. 8.3 months).

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