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Effects of metformin, rapamycin, and resveratrol on cellular metabolism of canine primary fibroblast cells isolated from large and small breeds as they age

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Abstract

Small breed dogs have longer lifespans than their large breed counterparts. Previous work demonstrated that primary fibroblast cells isolated from large breed young and old dogs have a persistent glycolytic metabolic profile compared with cells from small breed dogs. Here, we cultured primary fibroblast cells from small and large, young and old dogs and treated these cells with three commercially available drugs that show lifespan and health span benefits, and have been shown to reduce glycolytic rates: rapamycin (rapa), resveratrol (res) and metformin (met). We then measured aerobic and anaerobic cellular respiration in these cells. We found that rapa and res increased rates of non-glycolytic acidification in small and large breed puppies and basal oxygen consumption rates (OCR) in small and large breed puppies. Rapa increased proton leak and non-mitochondrial respiration in small and large breed puppies. Maximal respiration was significantly altered with rapa treatment but in opposing ways: large breed puppies showed a significant increase in maximal respiration when treated with rapa, and small old dogs demonstrated a significant decrease in maximal respiration when treated with rapa. In opposition to rapa treatments, met significantly decreased basal OCR levels in cells from small and large breed puppies. Our data suggest that rapa treatments may be metabolically beneficial to dogs when started early in life and more beneficial in larger breeds.

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Data Availability

Data is available in its raw form by contacting the corresponding author. After publication, this dataset will be deposited onto figshare.

Code availability

Code will be available on Dr. William Cipolli’s website or by contacting him directly via email.

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Acknowledgments

We are grateful to the following veterinarians and veterinary practices for providing us with samples: Dr. Kerri Hudson, Dr. James Gilchrist, Dr. Heather Culbertson and Morgan Peppenelli at Waterville Veterinary Clinic (New York); Dr. Frank Capella from Village Vet in Wampsville, NY. Pet Street Station Animal Hospital (New York); Dr. Jim Bader at Mapleview Animal Hospital (Michigan). We are also grateful to the following breeders for participating in our study: Rhonda Poe, Bob Stauffer, Allison Mitchell, Nancy Secrist, Valeria Rickard, Joanne Manning, Lita Long, Betsy Geertson, Susan Banovic, Lisa Uhrich, Sheryl Beitch, Al Farrier, Barbara Hoopes, and Rachel Sann.

Funding

The Seahorse XFe96 oxygen flux analyzer was purchased via a National Science Foundation Major Research Instrument grant (NSF MRI 1725841 to AGJ). A Research Council grant from Colgate University to AGJ partly funded this work.

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Authors and Affiliations

  1. Department of Biology, Colgate University, 13 Oak Dr, Hamilton, NY, 13346, USA

    Ana Gabriela Jimenez

  2. Department of Mathematics, Colgate University, 13 Oak Dr, Hamilton, NY, 13346, USA

    Sahil Lalwani & William Cipolli

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  1. Ana Gabriela Jimenez
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  2. Sahil Lalwani
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  3. William Cipolli
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Contributions

AGJ designed the experiments, collected tissue, grew cells, collected data, and wrote the first draft of the manuscript. SL and WC cleaned the raw data, performed the data analyses, wrote the supplement, and edited the manuscript.

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Correspondence to Ana Gabriela Jimenez.

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The authors declare no conflict of interest

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All the procedures within this study were approved by Colgate University's Institutional Care and Use Committee’s under protocol number 1819-13.

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Jimenez, A.G., Lalwani, S. & Cipolli, W. Effects of metformin, rapamycin, and resveratrol on cellular metabolism of canine primary fibroblast cells isolated from large and small breeds as they age. GeroScience 43, 1669–1682 (2021). https://doi.org/10.1007/s11357-021-00349-7

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