607 - RNA Sequencing of Wharton’s Jelly-derived Mesenchymal Stem Cells (MSCs) from Full-Term Healthy and Gestational Diabetes Pregnancies

Publication Number: 4594.607

Alexandria Mayne, Michigan State University College of Human Medicine, Lansing, MI, United States; Suraj Karanje, Michigan State University, Lansing, MI, United States; Amal Abdul-Hafez, Michigan State University, East Lansing, MI, United States; Ranga Prasanth Thiruvenkataramani, Michigan State University College of Human Medicine, Lansing, MI, United States; Hady Omar, Michigan State University College of Human Medicine, East Lansing, MI, United States; Burra V. Madhukar, Michigan State University College of Human Medicine, E. Lansing, MI, United States; Ahmed Abdelgawad, University of Delaware, Newark, DE, United States; Mona Batish, University of Delaware, Newark, DE, United States; Said Omar, Michigan State University College of Human Medicine, Lansing, MI, United States

Background: An innovative therapy is needed for the prevention of lung injury in premature neonates and the prevention of the development of bronchopulmonary dysplasia (BPD). In recent years, Mesenchymal Stem Cells (MSCs) from various tissues have emerged as promising sources for tissue regeneration. MSCs increase angiogenesis, reduce fibrosis, suppress inflammation, and modulate immune responses. Placenta may serve as a source of universal donor of MSCs and may have a therapeutic effect in preventing neonatal hyperoxia induced acute lung injury and development of BPD. Previously, we examined the characteristics of MSCs isolated from Healthy (H) full term pregnancies. Hyperglycemia, one of the most prominent features of gestational diabetes (GDM), may affect the proliferative profile and the biological and molecular characteristics of MSCs.

Objective: To examine the whole transcriptome sequencing and differential expression profiles of MSCs RNA isolated from WJ of the placentae of healthy and well controlled GDM pregnancies to elucidate the molecular mechanisms underlying biological functions, therapeutic effects and biomarker potentials of MSCs.

Design/Methods: Placentas were collected after IRBs approval and obtaining maternal informed consents. RNA was extracted from MSCs and was used for whole transcriptome sequencing. RNA was prepped with the KAPA RNA Hyperprep Kit w/ QIASeq FastSelect depletion and PR001839_OMAS_smRNA was prepped with the SMARTer smRNA-Seq Kit from Takara Bio.

Results: RNA sequencing data were obtained from 12 samples of N and GDM placental WJ derived MSCs at passages 2 and 4. Each RNAseq library has between 48-65M reads that passed QC, with 95% of the bases having quality scores of 30 or better. Hierarchical clustering analysis of RNAs with altered expression between the two groups is shown in Figure 1. There is no significant difference in the expression levels between normal and GDM MSCs of the upregulated and downregulated genes but there is minimal difference in the pathways they are enriched in as seen in Figure 2 and 3.

Conclusion(s): By using RNA sequencing, there was no major difference in gene expression between healthy and well controlled gestational diabetes WJ MSCs, and specifically no significant differentially expressed genes with regards to tissue regeneration and immune modulation.

PCA plot, Heat Map, and DEG
Hierarchical clustering analysis of mRNAs with altered gene expression between the two groups. Red strip, high relative expression; blue strip, low relative expression; white strip, no change in gene expression. Color intensity reflects the degree of expression (increase or decrease). Differentially expressed genes (DEG) with no significant difference in gene expression for MSCs mRNA from Gestational Diabetic vs normal MSCs. D: Isolated from placental WJ MSCs of Gestational Diabetes; W: Isolated from placental WJ MSCs of Normal Placenta.

Gene Ontology Analysis of genes for up and down regulated genes
Gene Ontology (GO) analysis showed up-regulated and down-regulated genes with no significant difference between normal and GDM MSCs.

Gene Ontology Analysis of genes for Tissue regeneration and Immune Modulation
Gene Ontology (GO) analysis showed no difference between normal and GDM expression in genes for tissue regeneration and immune modulation.