Eliseev Lab Roman A. Eliseev, M.D., Ph.D. View Eliseev Profile Ph.D. 2002 University of Rochester School of Medicine & Dentistry Assistant Professor of Orthopaedics Primary Appointment: Department of Orthopaedics Secondary Appointment: Pharmacology and Physiology Center Affiliation: Center for Musculoskeletal Research Research Overview The main goal of our research efforts is to understand how cell metabolism determines cell fate and how it can be manipulated for the purposes of prevention and therapies. Mitochondria are “powerhouses” of the cell and also important components of cell signaling. In undifferentiated stem cells mitochondria are inactive but become activated during the process of differentiation. We focus on the mechanism of this metabolic switch in mesenchymal stem cells (MSC), common precursors of osteoblasts, chondrocytes, and adipocytes. Our data and the literature indicate that activation of mitochondria is especially important for osteogenic differentiation of MSCs; and disruption of mitochondrial function in MSCs decreases osteogenic potential of these cells. Therefore, understanding the mechanism regulating mitochondrial function in MSCs and finding ways to improve and protect mitochondrial function in MSCs may lead to development of new strategies to improve bone health. While studying MSC function in aged mice showing osteoporosis, we have found that mitochondria in these MSCs are less functional due to the mitochondrial permeability transition (MPT), a non-selective mitochondrial pore regulated by cyclophilin D (CypD). Importantly, CypD knockout mice, a loss-of-function model of the MPT, have more viable and more potent MSCs, less osteoporosis and stronger bones even when they are aged. These data suggest that CypD can be an important target to treat aging-related osteoporosis and delayed fracture healing. In addition, we study how cell metabolism regulates sensitivity of osteosarcoma to treatments. Osteosarcoma is the most frequent primary bone cancer affecting mostly children and young adults. It is notoriously resistant to radiation and easily spreads to the lung where it becomes practically incurable. Similarly to undifferentiated stem cells, many cancer cells show preference for glycolytic over mitochondrial metabolism even in the presence of adequate oxygen, a phenomenon known as the Warburg effect. Our data indicate that the Warburg effect is especially pronounced in osteosarcoma cancer stem cells (CSC), and reversal of this effect makes CSCs less viable and more sensitive to radiation. Projects 1. Metabolic Changes in MSCs During Differentiation The goal of this project is to determine the mechanism of activation of mitochondria in MSCs during osteogenic differentiation. We study human MSCs and mouse MSCs isolated from bone marrow of either wild-type or genetically modified mice. MSC ability to self-renew and differentiate into various lineages (osteogenic, adipogenic and chondrogenic) is being investigated in the context of cell metabolism. We monitor how mitochondrial function, morphology, ultrastructure and proteome as well as cell metabolome and gene expression, change during differentiation into various lineages. We also use gain- and loss-of-function approach to determine what signaling systems are involved in regulation of cell metabolism during differentiation. 2. MSC Metabolism and Osteogenic Potential During Aging This project is aimed to determine the mechanism of mitochondrial dysfunction in MSCs during aging leading to their decreased osteogenicity, bone loss, and delayed fracture healing. Mitochondrial permeability transition (MPT), a non-selective mitochondrial pore regulated by cyclophilin D (CypD), has been documented as the mechanism of mitochondrial dysfunction in cardiovascular and other systems during aging. The role of the MPT in aged bone has not been elucidated. Our data indicate that the MPT loss of function in CypD knock-out mice leads to improved mitochondrial function and osteogenicity in aged MSCs and less osteoporosis when compared to aged wild type mice. The role of CypD as a potential target to treat osteoporosis and delayed fracture healing and the potential of stem cell therapy involving MSCs with modified CypD function, is being investigated. 3. Metabolic Reprogramming in Osteosarcoma We study how increased glycolytic metabolism and decreased mitochondrial function determine resistance of osteosarcoma cancer stem cells (CSC) to treatments. Our data indicate that CSCs show the most pronounced Warburg effect meaning that they are more glycolytic and less oxidative than the rest of the cells within the tumor. This phenomenon makes them very resistant to apoptosis induced by radiation or other treatments. It also opens a window of opportunity to specifically target these CSCs by shifting their metabolism from glycolytic towards oxidative. We have also found that function of anti- and pro-apoptotic members of the Bcl2 family proteins depends on the level of mitochondrial activity. Selected Publications Zuch D, Giang A, Shapovalov Y, Schwarz E, Rosier R, O’Keefe R, and RA Eliseev, "Targeting Radioresistant Osteosarcoma Stem Cells with Parthenolide", J Cell Biochem, 113, 1282-91, 2012 Shapovalov Y, Hoffman D, Zuch D, DeMesy-Bentley K, and RA Eliseev, “Mitochondrial dysfunction in cancer cells due to aberrant mitochondrial replication”, J Biol Chem, 286, 22331-8, 2011 Shapovalov Y, Benavidez D, Zuch D, and RA Eliseev, “Proteasome inhibition with bortezomib suppresses growth and induces apoptosis in osteosarcoma”, Intl J Cancer, 127, 67-76, 2010 Eliseev RA, Malecki J, Lester T, Zhang Y, Humphrey J, and TE Gunter, “Cyclophilin D interacts with Bcl2 and exerts an anti-apoptotic effect”, J Biol Chem, 284, 9692-9, 2009 Eliseev RA, Dong YF, Sampson E, Zuscik MJ, Schwarz EM, O'Keefe RJ, Rosier RN, and MH Drissi, “Runx2-mediated activation of the Bax gene increases osteosarcoma cell sensitivity to apoptosis”, Onco-gene, 27, 3605–14, 2008 Eliseev RA, Filippov G, Velos J, VanWinkle B, Goldman A, Rosier RN, and TE Gunter, “Role of cyclophilin D in the resistance of brain mitochondria to the permeability transition”, Neurobiol Aging, 28, 1532-42, 2007 Eliseev RA, Schwarz EM, Zuscik MJ, O’Keefe RJ, Drissi H, and RN Rosier, “Smad7 Mediates Inhibition of Saos2 Osteosarcoma Cell Differentiation by NF-kB”, Exp Cell Res,312, 40-50, 2006 Eliseev RA, Zuscik MJ, Schwarz EM, O’Keefe RJ, Drissi H, and RN Rosier, “Increased Radiation-induced Apoptosis of Saos2 Cells via Inhibition of NF?B: a Role for cJun N-terminal Kinase”, J Cell Biochem, 96, 1262-73, 2005 Eliseev RA, VanWinkle B, Rosier RN, and TE Gunter, “Diazoxide-Mediated Preconditioning Against Apoptosis Involves Activation of CREB and NF-kB”, J Biol Chem, 279, 46748-54, 2004 Gunter TE, Yule DI, Gunter KK, Eliseev RA, and JD Salter, “Calcium and Mitochondria”, FEBS Letters, 567, 96-102, 2004 Eliseev RA, Salter, J, Gunter, KK, and TE Gunter, “Bcl-2 and tBid Proteins Counter-regulate Mitochondrial Potassium Transport”, Biochem Biophys Acta, 45227, 1-5, 2003 Eliseev RA, Gunter, TE, and KK Gunter, "Bcl-2 Sensitive Mitochondrial Potassium Accumulation and Swelling in Apoptosis", Mitochondrion, 1(4), 361-70, 2002 Graduate Program Affiliation Pathways of Human Disease (PWD) Cellular and Molecular Pharmacology and Physiology The Eliseev Lab is currently recruiting Graduate Students.