Dataset Information

An insight into Exosomal Cargo Isolated from three tissue specific Mesenchymal Stem Cells: Implication in Translational Research

ABSTRACT: Adult Mesenchymal stem cells (MSCs) derived exosomes have recently gained importance as a cell-free therapy for several chronic and inflammatory diseases. It is believed that these exosomes contain several biologically active molecules (i.e. miRNAs, proteins) which help to cure the diseases. The majority of published reports to date that have investigated the secretome of MSCs have done so using canonical expansion of cell culture conditions. However, the microenvironment experience by MSC post administration into animal models and patients is strikingly different. Thus instead of using secretome, Exosomes purified from adipose tissue (AD), Wharton Jelly (WJ) and Bone marrow BM) are widely studied. However, the content of the exosomes may differ based on their sources from where they are originated. It remains unclear, however, which types of proteins are packaged into exosomes compared with the cells from which they are derived. To explore these exosomes as a cell-free therapy for specific diseases, it is therefore, important to get comprehensive knowledge of the bioactive molecules present in the exosomes. In this study, we have purified the exosomes from three different sources (AD, WJ, and BM) and using RNA-Seq approach, we have categorized the common and different microRNAs present in the exosomes.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Stem Cell, Cell Culture

SUBMITTER: Arup Banerjee

LAB HEAD: ARUP BANERJEE

PROVIDER: PXD020661 | Pride | 2023-04-26

REPOSITORIES: Pride

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Project description:Purpose: Most studies conducted towards understanding the differences among the adult sources of MSCs (BM-MSC and AD-MSC) and UC-MSCs have focused on their phenotypic properties which cannot provide an idea of the functional characteristics changing at the molecular level. Even NGS studies done before, each each study focuses on a different profiling methodology, which makes it difficult to establish a molecular equivalency among the various datasets. We undertake an integrative omics comparison between UC-MSC and two other popular sources of adult MSC (BM-MSC and AD-MSC), with the reasoning that a joint modeling of the changes occurring at the various functional levels will provide greater in-depth insight into the molecular heterogeneity related to different sources of MSCs. Methods: human Mesenchymal Stem Cell (MSC) mRNA cultured under basal conditions in Xeno-free media were subjected to deep sequencing. Three different MSC sourcers : adipose (AD-MSC), bone-marrow (BM-MSC) and Wharton-jelly umbilical cord (UC-MSC) were used and three donors from each tissues source along with three biological replicates for each donor were sequenced using the Illumina PE 150 platform. The sequence reads that passed quality filters Q30≥ 80% were analyzed using DESeq2. Results: Using an optimized data analysis workflow, we identified 24,804 transcripts among the three different sources of MSCs. Among these 2127 transcripts (~10%) were found to be differentially expressed among the three sources of MSC, with a\Log2FC| ≥2 and FDR <0.05. Hierarchical clustering of differentially expressed genes uncovered significant differences among the neonatal source (UC-MSC) and adult-MSCs (AD-MSC and BM-MSC). Altered gene expression profiles among the sources was also confirmed using a proteomics study of the same three sources of MSC with the same donors. Further, secretory immune-regulatory molecules which were confirmed to be differentially expressed among the MSC sources using the integrated transcriptomics-proteomics approach was validated using a Luminex cytokine profiling study. Conclusions: Our study represents the first detailed integrated analysis of differential gene expression profiling among source-specific human Mesenchymal Stem Cells using a multi-omics approach to validate the observed transcriptomic changes at both the translational as well as secretory levels. Our results offer a comhrehensive evaluation of the differences in immune signaling that exists among the different sourcrs of MSC based on their tissue of origina nd which subsequently plays a decisive role in determining the "regenrative siganture" of these MSCs based on the match between "immune-signature" and "disease type".

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An insight into Exosomal Cargo Isolated from three tissue specific Mesenchymal Stem Cells: Implication in Translational Research

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