Cassia auriculata: Aspects of Safety Pharmacology and Drug Interaction

Safety pharmacology studies help in identifying preclinical adverse drug reactions. We carried out routine safety pharmacology with focus on cardiovascular variables and pharmacokinetic herb-drug interaction studies on rats fed with standardized traditional hydro-alcoholic extract and technology-based supercritical extract of Cassia auriculata for 12 weeks. Our studies indicate that both these extracts are pharmacologically safe and did not show any significant adverse reactions at the tested doses. The traditional hydro-alcoholic extract did not show any significant effect on pharmacokinetics; however, the technology-based supercritical extract caused a significant reduction in absorption of metformin. Our results indicate the need to include pharmacokinetic herb-drug interaction studies as evidence for safety especially for technology-based extracts

Integrated transcriptomic and proteomic analysis of the molecular cargo of extracellular vesicles derived from porcine adipose tissue-derived mesenchymal stem cells

<div><p>Background</p><p>Mesenchymal stromal/stem cell (MSC) transplantation is a promising therapy for tissue regeneration. Extracellular vesicles (EVs) released by MSCs act as their paracrine effectors by delivering proteins and genetic material to recipient cells. To assess how their cargo mediates biological processes that drive their therapeutic effects, we integrated miRNA, mRNA, and protein expression data of EVs from porcine adipose tissue-derived MSCs.</p><p>Methods</p><p>Simultaneous expression profiles of miRNAs, mRNAs, and proteins were obtained by high-throughput sequencing and LC-MS/MS proteomic analysis in porcine MSCs and their daughter EVs (n = 3 each). TargetScan and ComiR were used to predict miRNA target genes. Functional annotation analysis was performed using DAVID 6.7 database to rank primary gene ontology categories for the enriched mRNAs, miRNA target genes, and proteins. STRING was used to predict associations between mRNA and miRNA target genes.</p><p>Results</p><p>Differential expression analysis revealed 4 miRNAs, 255 mRNAs, and 277 proteins enriched in EVs versus MSCs (fold change >2, p<0.05). EV-enriched miRNAs target transcription factors (TFs) and EV-enriched mRNAs encode TFs, but TF proteins are not enriched in EVs. Rather, EVs are enriched for proteins that support extracellular matrix remodeling, blood coagulation, inflammation, and angiogenesis.</p><p>Conclusions</p><p>Porcine MSC-derived EVs contain a genetic cargo of miRNAs and mRNAs that collectively control TF activity in EVs and recipient cells, as well as proteins capable of modulating cellular pathways linked to tissue repair. These properties provide the fundamental basis for considering therapeutic use of EVs in tissue regeneration.</p></div

Characterization of MSC-derived EVs.

<p>A: Transmission electron microscopy showing cultured MSCs releasing EVs. B: EVs express common EV (CD9, CD29, and CD63) and MSC (CD73 and CD105) markers. C: Size distribution of isolated EVs revealed a similar proportion of small microvesicles and exosomes.</p

Validation of RNAseq and proteomic analysis.

<p>Expression of the candidate miRNAs (miR-140-3p and miR-378), genes (SMAD2 and POU2F1), and proteins (C2 and TGFβ-1) was concordant with the RNAseq and proteomics findings. MSC-derived EVs (PKH26, red) were internalized by cultured of human umbilical vein endothelial cells (HUVECs) (CD31, green). DAPI DAPI = blue, nuclei. Incubation of HUVECs with EVs increased HUVEC expression of miR-140-3p, miR-378, SMAD2, POU2F1, C2, and TGFβ-1. *p<0.05 vs. MSCs, ‡p<0.05 vs. HUVECs.</p

Overview of experimental design and data analysis.

<p>Abdominal fat was collected from female domestic pigs, and mesenchymal stem cells (MSCs) and their daughter extracellular vesicles (EVs) isolated and characterized. mRNA and miRNA sequencing analysis and LC-MS/MS proteomic analysis were performed in both MSCs and EVs (n = 3 each). Differentially expression analysis was performed and EV-enriched miRNA, mRNA, and proteins identified. miRNA predicted targets were identified with TargetScan and ComiR. Functional annotation clustering analysis was performed using DAVID 6.7 database to obtain a ranking of primary gene ontology categories for the enriched mRNA, miRNA target genes, and proteins. Venn diagrams were used to visualize genes shared between each group and their interactions, and STRING to predict associations between mRNA and miRNA target genes.</p

CD38CD39 Population DEGs DS1 from A Population of Tumor-Infiltrating CD4⁺ T Cells Co-Expressing CD38 and CD39 Is Associated with Checkpoint Inhibitor Resistance

CD38/CD39 populations DEGs</p