Unknown,Transcriptomics,Genomics,Proteomics

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Genome-wide tissue gene expression profiling in Juvenile GÃ¶ttingen Minipigs with customized Agilent microarrays

ABSTRACT: Minipigs have favorable biological characteristics for juvenile toxicity studies to assess efficacy and safety of pediatric drug products. To evaluate at the molecular level the expression of drug targets, drug metabolism pathways or general maturation of organ transcriptomes in minipigs, we used Customized Agilent Micoarrays (Design-ID 050244) for genome-wide gene expression profiling on 9 different tissues from male and female juvenile GÃ¶ttingen minipigs aged 1 week to 24 months. The gene expression results analyzed in this study are further described in Heckel T. et al. (2015) Functional analysis and transcriptional output of the GÃ¶ttingen minipig genome. under submission Data of 72 different tissue samples from 9 tissues (cerebral cortex, hypothalamus, cerebellum, liver, kidney, spleen, bone marrow, testis, ovary) and 4 age groups in female (1 week, 4 weeks, 2 months, 4 months) and 5 age groups in male (1 week, 4 weeks, 2 months, 4 months, 2 years) Goettingen minipigs (Ellegaard) were measured. One tissue was measured in 8 technical replicates. The combination of data of female and male pigs could be seen as a kind of biological replicate for most genes.

ORGANISM(S): Sus scrofa

SUBMITTER: Tobias Heckel

PROVIDER: E-GEOD-71441 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Functional analysis and transcriptional output of the Göttingen minipig genome.

Heckel Tobias T Schmucki Roland R Berrera Marco M Ringshandl Stephan S Badi Laura L Steiner Guido G Ravon Morgane M Küng Erich E Kuhn Bernd B Kratochwil Nicole A NA Schmitt Georg G Kiialainen Anna A Nowaczyk Corinne C Daff Hamina H Khan Azinwi Phina AP Lekolool Isaac I Pelle Roger R Okoth Edward E Bishop Richard R Daubenberger Claudia C Ebeling Martin M Certa Ulrich U

BMC genomics 20151114

<h4>Background</h4>In the past decade the Göttingen minipig has gained increasing recognition as animal model in pharmaceutical and safety research because it recapitulates many aspects of human physiology and metabolism. Genome-based comparison of drug targets together with quantitative tissue expression analysis allows rational prediction of pharmacology and cross-reactivity of human drugs in animal models thereby improving drug attrition which is an important challenge in the process of drug ...[more]

PMID: 26573612

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Project description:Description Primary membranous nephropathy (MN) is an autoimmune kidney disease histomorphologically defined by subepithelial deposition of immune complexes in the glomeruli, and exhibits a high risk for end-stage kidney disease. Circulating antibodies against phospholipase A2 receptor 1 (PLA2R1) are detected in 70-80% of MN patients and correlate with treatment response and prognosis. Experimental proof that human PLA2R1-antibodies induce MN is still missing. Methods: In passive transfer experiments, minipigs received plasma or purified IgG from patients with PLA2R1-associated MN or from healthy controls. PLA2R1-antibodies and proteinuria were monitored using Western blot, ELISA and Coomassie staining. Kidney tissues were analyzed using immunohistochemistry, immunofluorescence, electron microscopy and proteomic analyses. Results: We show that minipigs, like humans, express PLA2R1 on podocytes. Human PLA2R1-antibodies bound to minipig PLA2R1 in-vitro and in-vivo. Passive transfer of human PLA2R1-antibodies, derived from patients with PLA2R1- associated MN, led to the development of histomorphologic characteristics of human early-stage MN in minipigs, activation of components of the complement cascade and induction of low levels of proteinuria. In the later phases of disease, development of an autologous phase of disease was observed. Conclusions: Applying a translational approach from humans to minipigs we show that human PLA2R1-antibodies are pathogenic, although in the heterologous phase of disease only low level proteinuria developed. Proteomic samples: Porcine glomeruli were isolated following previously published protocols known for human glomeruli, with slight modifications (Stahl et al., Kidney Int. 1984;26(1):30–34.). A total of six samples of sieved glomeruli from porcine kidney were used. 1. untreated_negative_control Experiment 1 (passive transfer of human PLA2R1 antibody from PLA2R1-antibody positive patient into minipigs) 2. exp_1_passive_antibody_rep1 Same experimental procedure in two animals (replicate 1) (treated with patient antibody) 3. exp_1_passive_antibody_rep2 Same experimental procedure in two animals (replicate 2) (treated with patient antibody) 4. exp_1_passive_antibody_control control animal for Pig A and B (treated with healthy antibody) Experiment 2 (passive transfer of human plasma from PLA2R1-antibody positive patient into minipigs) 5. exp_2_passive_plasma treated animal (treated with patient plasma) 6. exp_2_passive_plasma_control control animal (treated with healthy plasma)

Dataset Information

Genome-wide tissue gene expression profiling in Juvenile GÃ¶ttingen Minipigs with customized Agilent microarrays

Publications

Functional analysis and transcriptional output of the Göttingen minipig genome.

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