Additional file 1: of Preparing pharmacists to deliver a targeted service in hypertension management: evaluation of an interprofessional training program

Interview Guide. (PDF 16 kb

Sample table and description of data collected.

Sample table and description of data collected.</p

Nephrotoxicity image analysis of 3D tubuloids as a potential phenotypic screening tool.

(A) Na+/K+-ATPase signal intensity in relative intensity units (RIU) diminishes with increasing dose of known nephrotoxins at 24-hour post-dose compared to media (non-toxic) control, scale bars are 200 μm. (B) Image quantification of the 95th percentile Na+/K+-ATPase Tubuloid intensity with respect to concentration at 24 hours post-dose.</p

Most similar rat kidney profiles to 2D and 3D RPTEC-TERT1 (monolayer vs. tubuloid) obtained from TXG-MAPr application.

Most similar rat kidney profiles to 2D and 3D RPTEC-TERT1 (monolayer vs. tubuloid) obtained from TXG-MAPr application.</p

15 marker Luminex panel and abbreviations.

(DOCX)</p

96-well Luminex assay on cisplatin injured samples in 2D and 3D.

(A) Injury assay design in 2D and 3D compares cells in monolayer to tubuloids following incubation with cisplatin at pre-determined timepoints. (B) All targets with signal from a 15-plex assay (2 targets, beta-2-microglobulin and TFF3, were not detected) with respect to 2D or 3D samples, and concentration of cisplatin, over time emphasized that 3D tubuloids had higher detectable proteins indicating injury from the 3D tubuloids than the 2D monolayer (C) The highest signal biomarkers were A2M, CXCL1, IL-18, KIM-1, and Osteopontin.</p

Materials list.

(DOCX)</p

Tubuloid biomarkers and respective membrane location.

Tubuloid biomarkers and respective membrane location.</p

High throughput, high content imaging of tubuloids.

(A) Tubuloids cultured in 384-well plates were imaged via Preciscan algorithm (Opera Phenix, Perkin Elmer), wherein individual tubuloids of predetermined diameter, morphology, and quantity per-well were identified and captured in high resolution. (B) Tubuloids display several key biomarkers for proximal tubules including KIM-1, Aquaporin-1, Aquaporin-3, OCT2, OAT1, SGLT2, Na+/K+-ATPase, Integrin beta 1, Acetylated tubulin, Vimentin, Cytokeratin, Osteopontin and HMOX1, and exhibit hollow lumens. Polarity is evident for several markers, including primary cilia which face the interior lumen, as well as Na+/K+-ATPase and integrin beta 1 on the basolateral side.</p

3D tubuloids demonstrate higher similarity to human kidney tissue than 2D monolayer via RNASeq.

(A) Visualization of the RNASeq dataset separated by 3D vs 2D for all samples: Gene changes in 3D model against 2D model are shown; |log2FC| higher than 10 is set to 10 and -log10(padj) higher than 50 is set to 50. (B) Number of differentially expressed genes (padj < 0.05) against the base line (0hr) are shown; red bar: Up-regulated genes, blue bar: Down-regulated genes. (C) Comparison of the expression profiles: Expression profiles of 2D and 3D models, proximal tubules subtypes from human kidney scRNAseq (Kuppe et al.; subtypes: S1, S1.2.1, S1.2.2, S1.2.3, S3.1, S3.2, S3.3), and rats from the TG-GATES data set are compared. Compound treated and un-treated profiles are separated in the comparison and similarity score is the Spearman correlation of expression levels. Box plots are showing distribution of comparison of one profile from one data set against one profile from the other data set; notches indicate 95% confidence interval; the average of each dataset is represented by a dashed, colored line. Rat genes are converted to human genes and common genes in two sets are compared in the analysis. The highest similarity of any dataset was the 3D RNASeq to Kuppe. This indicates that the 3D dataset may be more closely relevant to human kidney tissue than 2D.</p