We utilized a combination of longitudinal topographic profiling and singular value decomposition-initiated multidimensional scaling (SVD-MDS) to identify genes involved in the progression to advanced hepatic fibrosis.

2D, two-dimensional; ACR, acute cellular rejection; BRCA1, breast cancer 1, early onset; CDKN3, cyclin-dependent kinase inhibitor 3; COL, collagen; DEGs, differentially expressed genes; HCC, hepatocellular carcinoma; HCV, hepatitis C virus; HLA, human leukocyte antigen; HSC, hepatic stellate cell; IPA, Daporinad cost ingenuity pathways analysis; ISGs, interferon-stimulated genes; LGALS3, galectin 3; MDS, multidimensional scaling; NS, nonstructural protein; OLT, orthotopic liver transplantation; SVD, singular value decomposition; UNP, uninfected normal pool; UWMC, University of Washington Medical Center. Additional detail regarding methods can be found in the Supporting Materials and Methods. Core needle liver biopsies were obtained from liver transplant patients at the University of Washington Medical Center (UWMC; Seattle, WA). All patients provided informed consent according to protocols approved by the Human Subject Pritelivir concentration Review Committee at the University of Washington. No donor organs were obtained from executed prisoners or other institutionalized persons. Microarray raw data were extracted using the Bioconductor

limma package28 and were median normalized. For interassay comparisons Methane monooxygenase and longitudinal analysis, the normalization using weighted negative second-order exponential error functions method was used for normalization.29 Differentially expressed genes have been identified using a fold-change–based z-test statistic (with a fold-change parameter of 1.2; P < 0.01). SVD-MDS dimensionality reduction and subsequent two-dimensional (2D) representations were obtained using the SVD-MDS method.6 Kruskal stress represents information loss resulting

from dimensionality reduction/representation as a fraction of total information. The geometric objects (i.e., transcriptomic data for individual genes in different samples at different times) are nonlinearly deformed (i.e., MDS), rotated into the principal nonlinear dimensions (i.e., SVD), and then projected onto the plane. Therefore, the 2D representation captures features of the geometric objects that would otherwise only be visible in a space of higher dimension. Because the nonlinearity is not uniform, this space of higher dimension is not exactly defined, but typically corresponds to a space of two to four dimensions higher than that of the visual representation. SVD-MDS performs better than hierarchical clustering in this setting because it accounts for several of the principal dimensions of the data. Longitudinal analysis was achieved using the same methodology as employed previously.