Amanda Fazzalari1, Giacomo Basadonna2, Alper Kucukural3, Kahraman Tanriverdi4, Milka Koupenova4, Natalie Pozzi5, Jahnavi Kakuturu5, Ann-Kristin U Friedrich5, Ron Korstanje6, Nicholas Fowler7, Jerrold L Belant7, Dean E Beyer8, Marjory B Brooks9, Eric W Dickson10, Meghan Blackwood11, Chris Mueller11, J Alexander Palesty5, Jane E Freedman4, Mitchell A Cahan12. 1. Department of Surgery, University of Massachusetts Medical School, Worcester, Massachusetts; The Stanley J. Dudrick Department of Surgery, Saint Mary's Hospital, Waterbury, Connecticut. 2. Department of Surgery, University of Massachusetts Medical School, Worcester, Massachusetts. 3. Bioinformatics Core, University of Massachusetts Medical School, Worcester, Massachusetts; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts. 4. Division of Cardiovascular Medicine, Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts. 5. The Stanley J. Dudrick Department of Surgery, Saint Mary's Hospital, Waterbury, Connecticut. 6. The Korstanje Lab, The Jackson Laboratory, Bar Harbor, Maine. 7. Camp Fire Program in Wildlife Conservation, State University of New York College of Environmental Science and Forestry, Syracuse, New York. 8. Department of Fisheries and Wildlife, College of Agriculture & Natural Resources, Michigan State University, East Lansing, Michigan; Michigan Department of Natural Resources, Marquette, Michigan. 9. Comparative Coagulation Section, Cornell University Animal Health Diagnostic Center, Ithaca, New York. 10. Department of Emergency Medicine, University of Massachusetts Medical School, Worcester, Massachusetts. 11. Mueller Lab for Gene Therapy, Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts. 12. Department of Surgery, University of Massachusetts Medical School, Worcester, Massachusetts. Electronic address: mitchell.cahan@umassmemorial.org.
Abstract
BACKGROUND: Hibernating American black bears have significantly different clotting parameters than their summer active counterparts, affording them protection against venous thromboembolism during prolonged periods of immobility. We sought to evaluate if significant differences exist between the expression of microRNAs in the plasma of hibernating black bears compared with their summer active counterparts, potentially contributing to differences in hemostasis during hibernation. MATERIALS AND METHODS: MicroRNA sequencing was assessed in plasma from 21 American black bears in summer active (n = 11) and hibernating states (n = 10), and microRNA signatures during hibernating and active state were established using both bear and human genome. MicroRNA targets were predicted using messenger RNA (mRNA) transcripts from black bear kidney cells. In vitro studies were performed to confirm the relationship between identified microRNAs and mRNA expression, using artificial microRNA and human liver cells. RESULTS: Using the bear genome, we identified 15 microRNAs differentially expressed in the plasma of hibernating black bears. Of these microRNAs, three were significantly downregulated (miR-141-3p, miR-200a-3p, and miR-200c-3p), were predicted to target SERPINC1, the gene for antithrombin, and demonstrated regulatory control of the gene mRNA expression in cell studies. CONCLUSIONS: Our findings suggest that the hibernating black bears' ability to maintain hemostasis and achieve protection from venous thromboembolism during prolonged periods of immobility may be due to changes in microRNA signatures and possible upregulation of antithrombin expression.
BACKGROUND: Hibernating American black bears have significantly different clotting parameters than their summer active counterparts, affording them protection against venous thromboembolism during prolonged periods of immobility. We sought to evaluate if significant differences exist between the expression of microRNAs in the plasma of hibernating black bears compared with their summer active counterparts, potentially contributing to differences in hemostasis during hibernation. MATERIALS AND METHODS: MicroRNA sequencing was assessed in plasma from 21 American black bears in summer active (n = 11) and hibernating states (n = 10), and microRNA signatures during hibernating and active state were established using both bear and human genome. MicroRNA targets were predicted using messenger RNA (mRNA) transcripts from black bear kidney cells. In vitro studies were performed to confirm the relationship between identified microRNAs and mRNA expression, using artificial microRNA and human liver cells. RESULTS: Using the bear genome, we identified 15 microRNAs differentially expressed in the plasma of hibernating black bears. Of these microRNAs, three were significantly downregulated (miR-141-3p, miR-200a-3p, and miR-200c-3p), were predicted to target SERPINC1, the gene for antithrombin, and demonstrated regulatory control of the gene mRNA expression in cell studies. CONCLUSIONS: Our findings suggest that the hibernating black bears' ability to maintain hemostasis and achieve protection from venous thromboembolism during prolonged periods of immobility may be due to changes in microRNA signatures and possible upregulation of antithrombin expression.