Kidney Week

Abstract: SA-PO0307

Measurement of Lipogenic Flux in Kidney Using Deuterium Oxide (D2O)

Session Information

• Diabetic Kidney Disease: Basic and Translational Science Advances - 2
  November 08, 2025 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Diabetic Kidney Disease

• 701 Diabetic Kidney Disease: Basic

Authors

• Kayampilly, Pradeep, Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, United States

Pradeep Kayampilly, Ph.D

• Arnipalli, Manikanta Swamy, Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, United States

Manikanta Swamy Arnipalli, PhD

• Dixon, Alethia J, Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, United States

Alethia J Dixon, MSc, PhD

• Gerlach, Gary F., Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, United States

Gary F. Gerlach, PhD

• Saum, Keith Louis, Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, United States

Keith Louis Saum, MD, PhD

• Roeser, Nancy F., Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, United States

Nancy F. Roeser

• Pennathur, Subramaniam, Division of Nephrology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, United States

Subramaniam Pennathur, MD

Background

Diabetes is the leading cause of end stage kidney disease in the United States. Previous studies in our laboratory using stable isotope have shown that hyperglycemia leads to increased de novo lipogenesis (DNL) in kidney cells. Though lipogenic ¹³C-substrates detect DNL with high sensitivity, quantification is complicated. Here we show that the stable isotope Deuterium (2H) can be used both in vitro and in vivo to overcome these challenges. D2O treatment enables labeling of most biomolecules making it ideal for multiomic study. Administration of D2O to biological systems is simple and safe for longer time periods.

Methods

We developed a method using LC/ /MS to measure 2H labeling in target metabolites. Deuterium incorporation is measured as mass shifts in the spectra (m+2, m+3). The lipogenic flux is measured as the percentage of 2H labeling in palmitate, the primary product of FASn.

Results

Time and dosimetry studies in HK-2 cells show that incubating cells in media with 25% D2O for 48 hours is optimum to study lipogenic flux. DNLactivity in human podocyte under normal (NG) and hyperglycemic (HG) conditions was examined by incubating cells in D2O media with 5.5 mM or 25 mM glucose for 48 hours. As we have shown in our previous studies using ¹³C isotope, cells grown under hyperglycemic conditions show an enhanced 2H labeling of palmitate suggesting increased lipogenic flux (Fig A). For in vivo lipogenic flux studies mice with pathological features of DN (BKS db/db) and control (db/+) were given D2O (20ul/g BW) IP at 0 hour and 24 hour then sacrificed at 48 hour. Total lipids were extracted from kidney cortex and 2H labeling was analyzed. Our results show higher 2H labeling in diabetic mice kidney cortex (m+3, m+4 and m+5) (Fig B) indicating increased lipogenic flux.

Conclusion

These results show that 2H labeling with LC/MS can be used to study lipogenic flux in target tissues both in vivo and in vitro.

Results of in vitro and in vivo Deuterium labeling in kidney cells.

Funding

• Private Foundation Support

Digital Object Identifier (DOI)

• doi: 10.1681/ASN.2025md8h453z