Creasingly enriched with D0,STA,BL,ARIA,ARIB (IL17 pathway gene-set FDR = 0.3 p = 0.011) supporting the relevance of the IL17 pathway in AR. To validate these findings, we investigated an independent microarray data-set from 21 STA, and 14 rejection renal allograft biopsy cases which was publicly available in Gene Expression Omnibus (GEO, GSE9493). Two-class GSA across the STA and biopsy confirmed AR (n = 10) showed significant enrichment of both gene-sets IL17-pathway and Th17 sets 1655472 in these AR (IL17pathway, FDR = 0.33, p-value = 0.011; Th17 gene-set,unsupervised Principal Title Loaded From File Component Analysis (PCA) and hierarchical clustering were performed for the IL17 pathway and Th17 gene-set genes in AR and STA (Partek Genomics Suite V.6.6, Partek Inc. USA). In case of multiple array probe-sets for the same gene, the probe-set with the lowest deviation (standard error of mean) across the AR and STA samples was used for the analyses. In cases where different probe-sets for AR and STA revealed lower deviation, both probe-sets were used. In addition we evaluated if there were any specific transcriptional changes in biopsies that were C4d+ AR (n = 3) versus C4d2 AR (n = 10).Drug discovery targeting IL17 pathway and IL17+ Thelper cells genes. Affymetrix unique probe-IDs for selectedIL17 pathway and Th17 gene-set genes were uploaded into MetaCoreTM, an interactive platform of biologically relevant data to integrate rejection specific genes with annotated functional data of gene-protein, protein-protein, and protein-compound interactions, together with compound and drug Title Loaded From File content, and gene disease relationships (MetaCoreTM; GeneGo, Thomson Reuters, St. Joseph, MI). The Drug Lookup feature in MetaCoreTM correlated input genes with deposited data from therapeutic, non-therapeutic, and secondary drug interactions that had an underlying data entry in PubMed. Resulting compounds were then filtered for directDrug Repositioning Fenofibrate for TransplantationFDR = 0.39, p-value = 0.026). Overall, there were 140 gene-sets common in both data-sets which were significantly enriched in AR (FDR = 0.5). As the publicly downloaded data-set also included 4 cases among their 14 rejection cases that were defined as borderline acute rejection, we added a quantitative GSA analysis which revealed that the IL17 pathway was significantly enriched following STA,BL,AR (FDR = 0.5, p-value 0.008). Due to the small numbers of BL cases in comparison to the numbers of STA and AR cases in this data-set, these analyses might be skewed. Additional QGSA across 10 gene-sets representing the transcriptome of innate and adaptive immune cells (Monocytes, Natural Killer Cells, Dendritic Cells, Th1-cells, Th2-cells, Th17cells, T-regulatory cells, gamma delta T-cells) in response to experimental activation, and T-cell differentiation (Th-Differentiation, Anergy/Regulation) identified enrichment of the IL17+ Thelper cell gene-set (Th17, 157 unique genes) [30] in increasing AR (FDR 0.1, p = 0.047; Table 2). The Th17 gene-set response was more redundant in the AR biopsies than the Th1 gene-set response [31] (FDR = 0.1, p = 0.05). Genes regulated in the IL17 pathway and the Th17 gene-sets allowed for almost complete separation of rejecting renal allograft biopsies from non-rejecting biopsies by hierarchical clustering (Euclidean Distance and Cosine Dissimilarity) (Figure 2A, 2B). A total of 8 genes (Il-17 pathway, n = 1; Th17 gene-set, n = 8) were significantly different expressed between C4d+ AR vs. C.Creasingly enriched with D0,STA,BL,ARIA,ARIB (IL17 pathway gene-set FDR = 0.3 p = 0.011) supporting the relevance of the IL17 pathway in AR. To validate these findings, we investigated an independent microarray data-set from 21 STA, and 14 rejection renal allograft biopsy cases which was publicly available in Gene Expression Omnibus (GEO, GSE9493). Two-class GSA across the STA and biopsy confirmed AR (n = 10) showed significant enrichment of both gene-sets IL17-pathway and Th17 sets 1655472 in these AR (IL17pathway, FDR = 0.33, p-value = 0.011; Th17 gene-set,unsupervised Principal Component Analysis (PCA) and hierarchical clustering were performed for the IL17 pathway and Th17 gene-set genes in AR and STA (Partek Genomics Suite V.6.6, Partek Inc. USA). In case of multiple array probe-sets for the same gene, the probe-set with the lowest deviation (standard error of mean) across the AR and STA samples was used for the analyses. In cases where different probe-sets for AR and STA revealed lower deviation, both probe-sets were used. In addition we evaluated if there were any specific transcriptional changes in biopsies that were C4d+ AR (n = 3) versus C4d2 AR (n = 10).Drug discovery targeting IL17 pathway and IL17+ Thelper cells genes. Affymetrix unique probe-IDs for selectedIL17 pathway and Th17 gene-set genes were uploaded into MetaCoreTM, an interactive platform of biologically relevant data to integrate rejection specific genes with annotated functional data of gene-protein, protein-protein, and protein-compound interactions, together with compound and drug content, and gene disease relationships (MetaCoreTM; GeneGo, Thomson Reuters, St. Joseph, MI). The Drug Lookup feature in MetaCoreTM correlated input genes with deposited data from therapeutic, non-therapeutic, and secondary drug interactions that had an underlying data entry in PubMed. Resulting compounds were then filtered for directDrug Repositioning Fenofibrate for TransplantationFDR = 0.39, p-value = 0.026). Overall, there were 140 gene-sets common in both data-sets which were significantly enriched in AR (FDR = 0.5). As the publicly downloaded data-set also included 4 cases among their 14 rejection cases that were defined as borderline acute rejection, we added a quantitative GSA analysis which revealed that the IL17 pathway was significantly enriched following STA,BL,AR (FDR = 0.5, p-value 0.008). Due to the small numbers of BL cases in comparison to the numbers of STA and AR cases in this data-set, these analyses might be skewed. Additional QGSA across 10 gene-sets representing the transcriptome of innate and adaptive immune cells (Monocytes, Natural Killer Cells, Dendritic Cells, Th1-cells, Th2-cells, Th17cells, T-regulatory cells, gamma delta T-cells) in response to experimental activation, and T-cell differentiation (Th-Differentiation, Anergy/Regulation) identified enrichment of the IL17+ Thelper cell gene-set (Th17, 157 unique genes) [30] in increasing AR (FDR 0.1, p = 0.047; Table 2). The Th17 gene-set response was more redundant in the AR biopsies than the Th1 gene-set response [31] (FDR = 0.1, p = 0.05). Genes regulated in the IL17 pathway and the Th17 gene-sets allowed for almost complete separation of rejecting renal allograft biopsies from non-rejecting biopsies by hierarchical clustering (Euclidean Distance and Cosine Dissimilarity) (Figure 2A, 2B). A total of 8 genes (Il-17 pathway, n = 1; Th17 gene-set, n = 8) were significantly different expressed between C4d+ AR vs. C.
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