title: Pro-Inflammatory Cytokines Predict Relapse-Free Survival after One Month of Interferon-α but Not Observation in Intermediate Risk Melanoma Patients.
abstract: BACKGROUND
  E1697 was a phase III trial of adjuvant interferon (IFN)-α2b for one month (Arm B) versus observation (Arm A) in patients with resected melanoma at intermediate risk. We evaluated the levels of candidate serum cytokines, the HLA genotype, polymorphisms of CTLA4 and FOXP3 genes and the development of autoantibodies for their association with relapse free survival (RFS) in Arm A and Arm B among 268 patients with banked biospecimens.
  
  
  METHODS
  ELISA was used to test 5 autoantibodies. Luminex/One Lambda LABTypeRSSO was used for HLA Genotyping. Selected CTLA4 and FOXP3 Single nucleotide polymorphisms (SNPs) and microsatellites were tested for by polymerase chain reaction (PCR). Sixteen serum cytokines were tested at baseline and one month by Luminex xMAP multiplex technology. Cox Proportional Hazards model was applied and the Wald test was used to test the marginal association of each individual marker and RFS. We used the Lasso approach to select the markers to be included in a multi-marker Cox Proportional Hazards model. The ability of the resulting models to predict one year RFS was evaluated by the time-dependent ROC curve. The leave-one-out method of cross validation (LOOCV) was used to avoid over-fitting of the data.
  
  
  RESULTS
  In the multi-marker modeling analysis conducted in Arm B, one month serum IL2Rα, IL-12p40 and IFNα levels predicted one year RFS with LOOCV AUC = 82%. Among the three markers selected, IL2Rα and IFNα were the most stable (selected in all the cross validation cycles). The risk score (linear combination of the 3 markers) separated the RFS curves of low and high risk groups well (p = 0.05). This model did not hold for Arm A, indicating a differential marker profile in Arm B linked to the intervention (adjuvant therapy).
  
  
  CONCLUSIONS
  Early on-treatment proinflammatory serum markers (IL2Rα, IL-12p40, IFNα) significantly predict RFS in our cohort of patients treated with adjuvant IFN-α2b and warrant further study.
authors.affiliation: University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Ahmad A
authors.initials: AA
authors.lastname: Tarhini
authors.affiliation: University of Pittsburgh Cancer Institute Biostatistics Facility, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Yan
authors.initials: Y
authors.lastname: Lin
authors.affiliation: University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Haris
authors.initials: H
authors.lastname: Zahoor
authors.affiliation: University of Pittsburgh Cancer Institute Biostatistics Facility, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Yongli
authors.initials: Y
authors.lastname: Shuai
authors.affiliation: University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Lisa H
authors.initials: LH
authors.lastname: Butterfield
authors.affiliation: University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Steven
authors.initials: S
authors.lastname: Ringquist
authors.affiliation: Hellenic Cooperative Oncology Group, Athens, Greece.
authors.forename: Helen
authors.initials: H
authors.lastname: Gogas
authors.affiliation: University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Cindy
authors.initials: C
authors.lastname: Sander
authors.affiliation: Dana Farber Cancer Institute, Boston, Massachusetts, United States of America.
authors.forename: Sandra
authors.initials: S
authors.lastname: Lee
authors.affiliation: St. Luke's Cancer Center, Bethlehem, Pennsylvania, United States of America; Temple University, Philadelphia, Pennsylvania, United States of America.
authors.forename: Sanjiv S
authors.initials: SS
authors.lastname: Agarwala
authors.affiliation: University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: John M
authors.initials: JM
authors.lastname: Kirwood
mesh_terms: D000328:Adult; D000368:Aged; D060908:CTLA-4 Antigen / Q000235:genetics; D016207:Cytokines / Q000097:blood*; D018572:Disease-Free Survival; D005260:Female; D051858:Forkhead Transcription Factors / Q000235:genetics; D005838:Genotype; D006680:HLA Antigens / Q000235:genetics; D006801:Humans; D016898:Interferon-alpha / Q000627:therapeutic use*; D008297:Male; D008545:Melanoma / Q000097:blood / Q000188:drug therapy* / Q000235:genetics; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide; D011379:Prognosis; D012878:Skin Neoplasms / Q000097:blood / Q000188:drug therapy* / Q000235:genetics
chemical_list: D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; D016207:Cytokines; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D006680:HLA Antigens; D016898:Interferon-alpha