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GM Food Allergy Biomarkers
1. Development of Biomarkers for the Detection of Potential Allergenicity of Novel Food Mainul Husain Department of Animal & Poultry Science University of Guelph Ontario, Canada
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7. Overall Project Design Validation of Candidate Bio-Marker Genes By Real Time RT- PCR Data Analysis Mapping Genes To Functional Pathways Allergen Treated Experimental Mice Control Mice Total RNA Extracted From Spleens Total RNA Extracted From Spleens Pooled Total RNA Pooled Total RNA SELECTION OF GENES AS ALLERGY BIO-MARKERS Affymetrix Microarray Classification of Genes Using Gene Ontology
8. Balb/c Mice Challenged With Egg Ovomucoid Protein Total 30 Female BALB/c Mice In Control and Treated Groups 15 Control Mice Challenged with Amino Acid Solution (1mg/100 l+10 g CT) 15 Experimental Mice Challenged with Ovomucoid (1mg/100 l+10 g CT) Group 1 3 Mice Group 2 3 Mice Group 3 3 Mice Group 4 3 Mice Group 5 3 Mice Group 1 3 Mice Group 2 3 Mice Group 3 3 Mice Group 4 3 Mice Group 5 3 Mice Pooled Total RNA Pooled Total RNA Pooled Total RNA Pooled Total RNA Pooled Total RNA Pooled Total RNA
13. KEGG Pathways Affected In Response to Ovomucoid Challenge Pathways Number of Genes Identified MAPK SIGNALING PATHWAY 27 OXIDATIVE PHOSPHORYLATION 24 FOCAL ADHESION 24 CYTOKINE-CYTOKINE RECEPTOR INTERACTION 24 REGULATION OF ACTIN CYTOSKELETON 20 WNT SIGNALING PATHWAY 19 CELL CYCLE 18 PURINE METABOLISM 16 INSULIN SIGNALING PATHWAY 15 PENTOSE AND GLUCURONATE INTERCONVERSIONS 13 PORPHYRIN AND CHLOROPHYLL METABOLISM 13 STARCH AND SUCROSE METABOLISM 13 METABOLISM OF XENOBIOTICS BY CYTOCHROME P450 13 T CELL RECEPTOR SIGNALING PATHWAY 13 JAK-STAT SIGNALING PATHWAY 13 HEMATOPOIETIC CELL LINEAGE 12 AXON GUIDANCE 12 ANDROGEN AND ESTROGEN METABOLISM 11 TGF-BETA SIGNALING PATHWAY 11 RIBOSOME 11 ADHERENS JUNCTION 10 PHOSPHATIDYLINOSITOL SIGNALING SYSTEM 10
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15. 15 Control Mice Treated with Amino Acid Solution (1mg/100 l + 10 g CT) C3H/HeJ Mice Challenged With Peanut Protein Total 30 C3H/HeJ Mice In Both Control and Treated Groups 15 Experimental Mice Challenged with Peanut Protein (1mg/100 l + 10 g CT) Group 1 3 Mice Group 2 3 Mice Group 3 3 Mice Group 4 3 Mice Group 5 3 Mice Group 1 3 Mice Group 2 3 Mice Group 3 3 Mice Group 4 3 Mice Group 5 3 Mice Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA Total RNA RNA Pool (5 random samples) RNA Pool (5 random samples) RNA Pool (5 random samples) RNA Pool (4 High Score Samples) RNA Pool (4 Low Score Samples) RNA Pool (4 High Score Samples) RNA Pool (3 Low Score Samples)
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17. Biological Classification of The Significant Genes Ontology ID Gene Ontology Terms # of Genes P-Value FDR 44238 Primary Metabolism 467 <0.0001 <0.0001 44237 Cellular Metabolism 475 <0.0001 <0.0001 43283 Biopolymer Metabolism 204 <0.0001 <0.0001 8152 Metabolism 504 <0.0001 <0.0001 7582 Physiological Process 705 0.0001 0.0023 43170 Macromolecule Metabolism 296 <0.0001 0.0025 50875 Cellular Physiological Process 650 0.0001 0.0029 6323 DNA Packaging 29 0.0002 0.0069 6325 Establishment and/or Maintenance of Chromatin Architecture 28 0.0003 0.0076 6259 DNA Metabolism 54 0.0006 0.0395 6139 Nucleobase, Nucleoside, Nucleotide And Nucleic Acid Metabolism 227 0.0007 0.0405 7001 Chromosome Organization and Biogenesis 29 0.0014 0.0700 16568 Chromatin Modification 20 0.0012 0.0705 6461 Protein Complex Assembly 18 0.0022 0.1158 8652 Amino Acid Biosynthesis 8 0.0037 0.1167 51244 Regulation of Cellular Physiological Process 215 0.0035 0.1176 51128 Regulation of Cell Organization and Biogenesis 10 0.0026 0.1186 6470 Protein Amino Acid Dephosphorylation 17 0.0025 0.1193 15031 Protein Transport 60 0.0027 0.1193 19538 Protein Metabolism 211 0.0034 0.1194
18. Directed Acyclic Graph (DAG) of Enriched Immune Response Category Some other significantly enriched sub-categories are: (1) Innate immune response, (2) Positive regulation of inflammatory response, (3) T-cell proliferation, (4) B-cell proliferation, (5) Positive regulation of interferon-beta biosynthesis.