Seromic analysis of antibody responses in non-small cell lung cancer patients and healthy donors using conformational protein arrays
Ludwig Institute for Cancer Research at Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA. Journal of Immunological Methods
(Impact Factor: 1.82).
12/2008; 341(1-2):50-8. DOI: 10.1016/j.jim.2008.10.016
Analysis of antibody responses to self-antigens has driven the development of the field of tumor immunology, with the identification of many protein targets found in cancer but with limited expression in normal tissues. Protein microarray technologies offer an unprecedented platform to assay the serological response of cancer patients to tumor antigens in a comprehensive fashion, against many proteins simultaneously. We developed an array containing 329 full-length proteins, originally identified as antigenic in various cancer patients by serological expression cloning (SEREX), that were immobilized as folded, functional products accessible for antibody binding. To validate the use of these microarrays, we selected 31 sera from non-small cell lung cancer patients previously known to react to the following antigens by ELISA: LAGE-1/CTAG2, MAGEA4, TP53, SSX and SOX2. These sera were compared with 22 sera from healthy donors for reactivity against a series of antigens present on microarrays. The sensitivity and specificity of the arrays compared favorably with standard ELISA techniques (94% concordance). We present here a stringent strategy for data analysis and normalization that is applicable to protein arrays in general, and describe findings suggesting that this approach is suitable for defining potential antigenic targets for cancer vaccine development, serum antibody signatures with clinical value, characterization of predictive serum markers for experimental therapeutics, and eventually for the serological definition of the cancer proteome (seromics).
Available from: Emilio Bria
- "In the actual study we sought to evaluate the subtypes of genotypic abnormalities of the entire chromosome 3 and the distal locus specific 3q that maps the SOX-2 and PI3CA genes in a serie of squamous lung carcinoma , , , , , , , , by weighting different chromosomal anomalies mapped by fluorescent ISH (FISH) and aCGH front techniques on routinely available formalin-fixed neoplastic tissue. "
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ABSTRACT: Squamous lung carcinoma lacks specific "ad hoc" therapies. Amplification of chromosome 3q is the most common genomic aberration and this region harbours genes having role as novel targets for therapeutics. There is no standard definition on how to score and report 3q amplification. False versus true 3q chromosomal amplification in squamous cell lung carcinoma may have tremendous impact on trials involving drugs which target DNA zones mapping on 3q. Forty squamous lung carcinomas were analyzed by FISH to assess chromosome 3q amplification. aCGH was performed as gold-standard to avoid false positive amplifications. Three clustered patterns of fluorescent signals were observed. Eight cases out of 40 (20%) showed ≥8 3q signals. Twenty out of 40 (50%) showed from 3 to 7 signals. The remaining showed two fluorescent signals (30%). When corrected by whole chromosome 3 signals, only cases with ≥8 signals maintained a LSI 3q/CEP3 ratio >2. Only the cases showing 3q amplification by aCGH (+3q25.3-3q27.3) showed ≥8 fluorescent signals at FISH evidencing a 3q/3 ratio >2. The remaining cases showed flat genomic portrait at aCGH on chromosome 3. We concluded that: 1) absolute copy number of 3q chromosomal region may harbour false positive interpretation of 3q amplification in squamous cell carcinoma; 2) a case results truly "amplified for chromosome 3q" when showing ≥8 fluorescent 3q signals; 3) trials involving drugs targeting loci on chromosome 3q in squamous lung carcinoma therapy have to consider false versus true 3q chromosomal amplification.
PLoS ONE 12/2012; 7(12):e49689. DOI:10.1371/journal.pone.0049689 · 3.23 Impact Factor
Available from: umich.edu
- "Around 30% of patients with lung adenocarcinoma exhibit a humoral response to glycosylated annexins I and/or II whereas none of the noncancerous standards exhibit such a response . In PDAC, auto-antibodies to DEAD-box protein 48 were observed in 33.3% of pancreatic patient sera, while none of the patients with benign disease and healthy controls showed reactivity against the antigen . MUC1  , p53  and Rad51  have also shown restricted immune reactivity in a subset of cancer samples. "
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ABSTRACT: The immunogenic nature of cancer can be explored to distinguish pancreatic cancer from related non-cancer conditions. We describe a liquid-based microarray approach followed by statistical analysis and confirmation for discovery of auto-immune biomarkers for pancreatic cancer. Proteins from the Panc-1 pancreatic cancer cell line were fractionated using a 2-D liquid separation method into over 1052 fractions and spotted onto nitrocellulose coated glass slides. The slides were hybridized with 37 pancreatic cancer sera, 24 chronic pancreatitis sera and 23 normal sera to detect elevated levels of reactivity against the proteins in spotted fractions. The response data obtained from protein microarrays was first analyzed by Wilcoxon Rank-Sum Tests to generate two lists of fractions that positively responded to the cancer sera and showed p-values less than 0.02 in the pairwise comparison between cancer specimens and normal and chronic pancreatitis specimens. The top 3 fractions with the lowest correlations were combined in receiver operating characteristic analyses. The area-under-the-curve (AUC) values are 0.813 and 0.792 for cancer vs. normal and cancer vs. pancreatitis respectively. Outlier-Sum statistics were then applied to the microarray data to determine the existence of outliers exclusive in cancer sera. The selected fractions were identified by LC-MS/MS. We further confirmed the occurrence of outliers with three proteins among cancer samples in a confirmation experiment using a separate dataset of 165 serum samples containing 48 cancer sera and 117 non-cancer controls. Phosphoglycerate kinase 1 (PGK1) elicited greater reactivity in 20.9% (10 in 48) of the samples in the cancer group, while no outlier was present in the non-cancer groups.
Cancer biomarkers: section A of Disease markers 01/2010; 7(1):25-37. DOI:10.3233/CBM-2010-0145 · 1.72 Impact Factor
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ABSTRACT: Summary form only given. High-voltage (HV) power transformers are exposed to a large variety of electric stresses during final testing as well as in service. In addition to the dielectric strength requirement, the insulation system must meet further requirements. Thermal and mechanical aspects also have to be taken into consideration when dimensioning the insulation. In consequence, oil ducts of different sizes and volumes are exposed to different types of stresses. The approach taken by the transformer industry to the design of their products for safe operation has been considered
Conduction and Breakdown in Dielectric Liquids,1993., ICDL '93., IEEE 11th International Conference on; 08/1993
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