Abstract: FR-PO946
Transcriptome Analyses of Altered Mouse Kidney Development Following Maternal Interleukin-6 Exposure During Gestation
Session Information
- Development, Stem Cells, Regenerative Medicine - II
October 26, 2018 | Location: Exhibit Hall, San Diego Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 501 Development, Stem Cells, and Regenerative Medicine: Basic
Authors
- Srivastava, Tarak, Childrens's Mercy Hospital, Kansas City, Missouri, United States
- Heruth, Daniel P., Childrens's Mercy Hospital, Kansas City, Missouri, United States
- Garola, Robert E., Childrens's Mercy Hospital, Kansas City, Missouri, United States
- Sharma, Mukut, KCVA Medical Center, Kansas City, Missouri, United States
Background
Children born to obese mothers have higher incidence of renal anomalies. Obesity is characterized by systemic inflammation. IL-6, an inflammatory cytokine, can cross the placental barrier, and is elevated in the amniotic fluid of obese women. Increasing evidence suggest that epigenetic mechanisms and microRNA (miRNA) interact in a bidirectional manner, a potential key mechanism for environmental effects on development. We hypothesized that renal development would be affected by elevated levels of maternal IL-6 brought about by epigenetic changes that alter both miRNA and RNA expression.
Methods
Pregnant C57BL/6 mice received either normal saline or IL-6 (10 pg/g BW) intraperitoneally on alternate days from E12.5. Fetal kidneys (E20.5) were removed from uterine horns by dissection and fixed in 10% formalin or used to isolate DNA and total RNA. LC-MS analysis of hydrolyzed DNA was performed to evaluate for methylation changes. TruSeq RNA Libraries (saline n=5, IL-6 n=5) were subjected to miRNA- and RNA-seq on an Illumina platform. Reads were mapped to the mouse reference genome and analyzed by miRDeep2.pl and Tuxedo Suite for miRNA-seq and RNA-seq, respectively.
Results
Maternal IL-6 administration resulted in decreased renal cortical parenchyma (358.4±74.3μm) compared with control (453.9±80.5μm, n=5, p=0.04). LC-MS analysis found a 10% increase in methylated cytosine in the IL-6 group suggesting epigenetic modifications in fetal kidney DNA. These modifications correlated (q<0.05) with 58 and 2087 differentially expressed (DE) miRNAs and RNAs, respectively. Stringent target prediction (score>90) identified 1350 mRNA regulated by the 58 miRNA of which 196 (9.4%) were present in the list of 2087 DE genes. Nephrotoxicity, as predicted by Ingenuity Pathway Analysis, associated with Renal Necrosis/Cell Death (p=1.5E-11, 104 genes) and Renal Damage (p=1.2E-8, 52 genes). We have identified the STAT3 (p=3.2E-06, 20 genes), NRF2-Mediated Oxidative Stress Response (p=3.5E-06, 37 genes) and Glucocorticoid Receptor Signaling (1.9E-05, 53 genes) pathways for further analysis and confirmation.
Conclusion
Our model of maternal IL-6 administration has allowed us to better understand the role of environment mediated changes on epigenetics and global gene expression in kidney development.
Funding
- Private Foundation Support