Kidney Week

Abstract: SA-PO1061

Placental Extracellular Vesicles from Preeclampsia Alter the Transcriptome of Murine Renal Proximal Tubule Epithelial Cells

Session Information

• Women's Health and Kidney Diseases
  October 26, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Women's Health and Kidney Diseases

• 2200 Women's Health and Kidney Diseases

Authors

• Pantham, Priyadarshini, University of California San Diego, La Jolla, California, United States

Priyadarshini Pantham, PhD

• Chousal, Jennifer N., University of California San Diego, La Jolla, California, United States

Jennifer N. Chousal, MD, PhD

• Ostrander, Tyler, University of California San Diego, La Jolla, California, United States

Tyler Ostrander

• Lindsay, Scott Alexander, University of California San Diego, La Jolla, California, United States

Scott Alexander Lindsay, PhD

• Edlabadkar, Anushka Ranjeet, University of California San Diego, La Jolla, California, United States

Anushka Ranjeet Edlabadkar

• Cheung, Willi, University of California San Diego, La Jolla, California, United States

Willi Cheung, MSc

• Bolisetty, Subhashini, University of Alabama at Birmingham, Birmingham, Alabama, United States

Subhashini Bolisetty, PhD

• Singh, Prabhleen, University of California San Diego, La Jolla, California, United States

Prabhleen Singh, MD, MPH

Background

Preeclampsia (PE), a deadly hypertensive disorder of pregnancy, is associated with maternal kidney damage indicated by proteinuria, increased risk of chronic kidney disease, and cardiorenal syndrome. The placenta plays a key role in the etiology of PE and releases extracellular vesicles (EVs) containing RNA cargo into the maternal circulation throughout pregnancy, which can modulate distant organs including the maternal kidney. The role of renal proximal tubule epithelial cells (rPTECs) in the etiology of kidney damage and their interaction with placental EVs in PE has never been studied. We aimed to characterize the effect of human placental EVs isolated from PE pregnancies compared to normal pregnancies on the transcriptome of murine rPTECs.

Methods

Placental EVs (70-100nm) were isolated using size exclusion chromatography following placental explant culture from normal (n=3) or preeclamptic (n=3) pregnancies. Murine rPTECs were isolated from non-pregnant female mouse kidneys (n=2) and treated with normal or PE placental EVs on day 7 at a dose of 1x10¹⁰EVs/mL for 24 hours. RNA was isolated from murine rPTECs, RNA libraries were constructed using the KAPA RNA HyperPrep Kit, and RNA-Seq was performed (Illumina NovaSeq X Plus). Reads were aligned using STAR, gene abundances quantified using RSEM, and differential expression analysis was conducted using DESeq2.

Results

An average of 41x10⁶ reads per sample was obtained, and >5000 transcripts were dysregulated in murine rPTECs treated with EVs from PE compared to normal pregnancies (FDR<0.001, Figure 1). Analysis of molecular pathways using gProfiler showed that these transcripts were significantly enriched for mRNAs involved in oxidative phosphorylation (42 mRNAs, FDR<0.01).

Conclusion

We have shown that placental EVs isolated from PE pregnancies alter the transcriptome of murine rPTECs, particularly in transcripts involved in mitochondrial electron transport. Current efforts are aimed at measuring mitochondrial oxygen consumption in proximal tubules isolated from pregnant mice injected with placental EVs from PE and normal pregnancies, to establish whether increased oxidative stress in rPTECs plays a key role in inducing the kidney damage seen in PE.

Funding

• Other NIH Support