Molecular models.

<p>Schematic representation of the constructed DN molecular model and the tacrolimus MoA molecular model with the list of significantly enriched molecular pathways based on the two molecular model features sets. Molecular pathways in bold font within the red border were significantly enriched in molecular features of both molecular model sets. Each box in the molecular model representations depicts one process unit, characterized by highly inter-connected proteins, with individual nodes representing protein coding genes.</p

Interference network.

<p>Depicted is the diabetic nephropathy molecular model with molecular features affected by tacrolimus being displayed with larger nodes and annotated with the official Gene Symbols. Molecular features upregulated by tacrolimus are highlighted in red, with downregulated molecular features displayed in green. Molecular features with contradictory evidence, i.e. up- and down-regulated are depicted in yellow. Annotated grey nodes depict molecular features being associated with effects of tacrolimus, in most cases SNPs in these genes having impact on tacrolimus efficacy. Prognostic and diagnostic biomarkers are indicated by a P or D respectively.</p

Months to equal risk compared between dialysis treatment and conservative management.

<p>Months to equal risk compared between dialysis treatment and conservative management.</p

Data_Sheet_1_The Effect of FGF23 on Cardiac Hypertrophy Is Not Mediated by Systemic Renin-Angiotensin- Aldosterone System in Hemodialysis.docx

Fibroblast growth factor 23 (FGF23) is elevated in patients with chronic kidney disease and contributes to left ventricular hypertrophy (LVH). The aim of the analysis was to determine whether this effect is mediated by the renin-angiotensin-aldosterone system (RAAS) in hemodialysis. Serum samples from 62 randomized hemodialysis patients with LVH were analyzed for plasma renin activity (PRA-S), angiotensin II (AngII), and metabolites, angiotensin-converting enzyme-2 (ACE2) and aldosterone using a high throughput mass spectrometry assay. Compared to healthy individuals, levels of the RAAS parameters PRA-S, AngII and aldosterone were generally lower [median (IQR) PRA-S 130 (46–269) vs. 196 (98, 238) pmol/L; AngII 70 (28–157) vs. 137 (76, 201) pmol/L; Aldosterone 130 (54, 278) vs. 196 (98, 238) pmol/L]. We did not find an indication that the effect of FGF23 on LVH was mediated by RAAS parameters, with all estimated indirect effects virtually zero. Furthermore, FGF23 was not associated with RAAS parameter levels throughout the study. While there was a clear association between FGF23 levels and left ventricular mass index (LVMI) at the end of the study and in the FGF23 fold change and LVMI change analysis, no association between RAAS and LVMI was observed. Serum concentrations of PRA-S, AngII, and aldosterone were below the ranges measured in healthy controls suggesting that RAAS is not systemically activated in hemodialysis patients. The effect of FGF23 on LVMI was not mediated by systemic RAAS activity. These findings challenge the current paradigm of LVH progression and treatment with RAAS blockers in dialysis.Clinical Trial Registration[https://clinicaltrials.gov/ct2/show/NCT03182699], identifier [NCT03182699].</p

MOESM1 of Effect of etelcalcetide on cardiac hypertrophy in hemodialysis patients: a randomized controlled trial (ETECAR-HD)

Additional file 1. SPIRIT 2013 Checklist: Recommended items to address in a clinical trial protocol and related documents

Whiskers denote 95% confidence intervals derived from linear mixed models adjusted for age, sex, and principal components of genetic variation.

<p>Whiskers denote 95% confidence intervals derived from linear mixed models adjusted for age, sex, and principal components of genetic variation.</p

A Gene Variant in CERS2 Is Associated with Rate of Increase in Albuminuria in Patients with Diabetes from ONTARGET and TRANSCEND

<div><p>Although albuminuria and subsequent advanced stage chronic kidney disease are common among patients with diabetes, the rate of increase in albuminuria varies among patients. Since genetic variants associated with estimated glomerular filtration rate (eGFR) were identified in cross sectional studies, we asked whether these variants were also associated with rate of increase in albuminuria among patients with diabetes from ONTARGET and TRANSCEND—randomized controlled trials of ramipril, telmisartan, both, or placebo. For 16 genetic variants associated with eGFR at a genome-wide level, we evaluated the association with annual rate of increase in albuminuria estimated from urine albumin:creatinine ratio (uACR). One of the variants (rs267734) was associated with rate of increase in albuminuria. The annual rate of increase in albuminuria among risk homozygotes (69% of the study population) was 11.3% (95%CI; 7.5% to 15.3%), compared with 5.0% (95%CI; 3.3% to 6.8%) for heterozygotes (27% of the population), and 1.7% (95%CI; −1.7% to 5.3%) for non-risk homozygotes (4% of the population); P = 0.0015 for the difference between annual rates in the three genotype groups. These estimates were adjusted for age, sex, ethnicity, and principal component of genetic heterogeneity. Among patients without albuminuria at baseline (uACR<30 mg/g), each risk allele was associated with 50% increased risk of incident albuminuria (OR = 1.50; 95%CI 1.15 to 1.95; P = 0.003) after further adjustment for traditional risk factors including baseline uACR and eGFR. The rs267734 variant is in almost perfect linkage-disequilibrium (r² = 0.94) with rs267738, a single nucleotide polymorphism encoding a glutamic acid to alanine change at position 115 of the ceramide synthase 2 (CERS2) encoded protein. However, it is unknown whether CERS2 function influences albuminuria. In conclusion, we found that rs267734 in CERS2 is associated with rate of increase in albuminuria among patients with diabetes and elevated risk of cardiovascular disease.</p></div

Association between SNPs and change in albuminuria.

<p>Results were adjusted for age, sex, 10 largest principal components of genetic variation and self-reported ethnicity (for “all ethnic groups” results).</p><p>Association between SNPs and change in albuminuria.</p

Regional plot for the CERS2 locus.

<p>SNPs are plotted by association P value of linear mixed models adjusted for age, sex, and principal components of genetic variation for the association between SNP and annual rate of change in albuminuria. and genomic position (NCBI Build 36). The original hit (rs267734) is labeled. The magnitude of linkage disequilibrium (r²) between each SNP and rs267734 is indicated by the intensity of the red coloring. Estimates of recombination rates are shown by the blue line. Gene positions are indicated by green arrows. Gene names are labeled. Linkage disequilibrium and recombination rates were estimated from the Utah residents of Northern and Western European ancestry (CEU) HapMap population (release 22). Plots were prepared using SNAP <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0106631#pone.0106631-Laviad1" target="_blank">[22]</a>. Panel A: P values adjusted for rs267734. Panel B: P values not adjusted for rs267734.</p

DataSheet_1_Effects of Reduced-Dose Anti-Human T-Lymphocyte Globulin on Overall and Donor-Specific T-Cell Repertoire Reconstitution in Sensitized Kidney Transplant Recipients.docx

BackgroundPre-sensitized kidney transplant recipients have a higher risk for rejection following kidney transplantation and therefore receive lymphodepletional induction therapy with anti-human T-lymphocyte globulin (ATLG) whereas non-sensitized patients are induced in many centers with basiliximab. The time course of lymphocyte reconstitution with regard to the overall and donor-reactive T-cell receptor (TCR) specificity remains elusive.Methods/DesignFive kidney transplant recipients receiving a 1.5-mg/kg ATLG induction therapy over 7 days and five patients with 2 × 20 mg basiliximab induction therapy were longitudinally monitored. Peripheral mononuclear cells were sampled pre-transplant and within 1, 3, and 12 months after transplantation, and their overall and donor-reactive TCRs were determined by next-generation sequencing of the TCR beta CDR3 region. Overall TCR repertoire diversity, turnover, and donor specificity were assessed at all timepoints.ResultsWe observed an increase in the donor-reactive TCR repertoire after transplantation in patients, independent of lymphocyte counts or induction therapy. Donor-reactive CD4 T-cell frequency in the ATLG group increased from 1.14% + -0.63 to 2.03% + -1.09 and from 0.93% + -0.63 to 1.82% + -1.17 in the basiliximab group in the first month. Diversity measurements of the entire T-cell repertoire and repertoire turnover showed no statistical difference between the two induction therapies. The difference in mean clonality between groups was 0.03 and 0.07 pre-transplant in the CD4 and CD8 fractions, respectively, and was not different over time (CD4: F(1.45, 11.6) = 0.64 p = 0.496; CD8: F(3, 24) = 0.60 p = 0.620). The mean difference in R20, a metric for immune dominance, between groups was -0.006 in CD4 and 0.001 in CD8 T-cells and not statistically different between the groups and subsequent timepoints (CD4: F(3, 24) = 0.85 p = 0.479; CD8: F(1.19, 9.52) = 0.79 p = 0.418).ConclusionReduced-dose ATLG induction therapy led to an initial lymphodepletion followed by an increase in the percentage of donor-reactive T-cells after transplantation similar to basiliximab induction therapy. Furthermore, reduced-dose ATLG did not change the overall TCR repertoire in terms of a narrowed or skewed TCR repertoire after immune reconstitution, comparable to non-depletional induction therapy.</p