Serum Protein Profiling to Identify High-Risk Neuroblastoma: Preclinical Relevance of Blood-Based Biomarkers

Sandoval, John A.; Turner, Katharyn E.; Hoelz, Derek J.; Rescorla, Frederick J.; Hickey, Robert J.; Malkas, Linda H.

doi:10.1016/j.jss.2007.03.058

« Previous Next »Journal of Surgical Research
Volume 142, Issue 2 , Pages 268-274, October 2007

Serum Protein Profiling to Identify High-Risk Neuroblastoma: Preclinical Relevance of Blood-Based Biomarkers

John A. Sandoval, M.D.
- Department of Surgery, Department of Medicine, Indiana University, School of Medicine and Riley Hospital for Children, Indianapolis, Indiana
Katharyn E. Turner, B.A.
- Section of Pediatric Surgery, Department of Medicine, Indiana University, School of Medicine and Riley Hospital for Children, Indianapolis, Indiana
Derek J. Hoelz, Ph.D.
- Division of Hematology/Oncology, Department of Medicine, Indiana University, School of Medicine and Riley Hospital for Children, Indianapolis, Indiana
Frederick J. Rescorla, M.D.
- Department of Surgery, Department of Medicine, Indiana University, School of Medicine and Riley Hospital for Children, Indianapolis, Indiana
- Section of Pediatric Surgery, Department of Medicine, Indiana University, School of Medicine and Riley Hospital for Children, Indianapolis, Indiana
Robert J. Hickey, Ph.D.
- Division of Hematology/Oncology, Department of Medicine, Indiana University, School of Medicine and Riley Hospital for Children, Indianapolis, Indiana
Linda H. Malkas, Ph.D.
- Division of Hematology/Oncology, Department of Medicine, Indiana University, School of Medicine and Riley Hospital for Children, Indianapolis, Indiana
- To whom correspondence and reprint requests should be addressed at Division of Hematology/Oncology, Indiana University Cancer Research Institute, 1044 West Walnut St, R4-202, Indianapolis, IN.

Received 8 January 2007 published online 30 August 2007.

Introduction

Development of early detection assays for advanced stage neuroblastoma (NB) remains elusive. We have previously shown that serum protein profiling technologies can differentiate healthy from NB children. As various sources of patient related bias exist in serum proteins, we hypothesized a well controlled animal model may provide a better method to identify tumor blood-based markers during NB progression.

Methods

Tumors were induced in the left kidneys of nude mice by the injection of cultured human NB cells (10⁶). Sera were collected from control and tumor-bearing mice at 2, 4, and 6 wk. Albumin-depleted sera were subjected to comparative proteomic profiling using 2D gel electrophoresis. Paired samples at each time point were analyzed and differentially expressed serum proteins were identified by mass spectrometry. Additionally, sera proteomic analysis from children with Stage IV NB and healthy controls were performed.

Results

Overexpression of five mouse serum proteins [α₁-acid glycoprotein, α₁-antitrypsin, α₂-macroglobulin, serum amyloid P-component, and serum amyloid A) were found only in NB-bearing mice. Changes in protein abundance were found to increase 2.5-fold (P ≤ 0.05) between 2-, 4-, and 6-wk old mice. Underexpression of immunoglobulin κ chain constant region was observed in the sera of tumor bearing mice compared with controls (2.5-fold, P ≤ 0.05). Among NB patients, α₁-acid glycoprotein, apolipoprotein A-IV, haptoglobin, and serum amyloid A were found to be up-regulated.

Conclusions

We identified distinct acute phase proteins that show up-regulation in both an animal tumor model and high-risk NB patients. As these serum proteins have been recognized as markers of tumor progression and prognosis in human malignancies, the validation of these polypeptides may enable serum proteomic profiling to become a valuable tool for identifying high-risk NB.

Key Words: neuroblastoma, high-risk pediatric malignancy, animal model, serum proteomics, acute phase proteins/response