Dr. Elissavet Kardami
Principal Investigator
Muscle Cell Biochemistry, Institute of Cardiovascular Sciences
Professor
Human Anatomy & Cell Science, University of Manitoba
Research Focus
(1) The role of FGF-2 in Acute and Chronic Cardiac Response to Injury
The heart expresses the growth factor FGF-2 (fibroblast growth factor-2) at all developmental stages. FGF-2 is made as high molecular weight (hmw) and low molecular weight (lmw) forms. We have studied the properties of these different forms and have established that lmw FGF-2 is very potent in raising cardiac and cellular resistance to injury and cell death, acting both as a ‘pre-conditioning’ and as a post-conditioning’ agent, in the acute cardiac injury setting, in culture and in vivo. LmwFGF-2 is also capable of sustained beneficial effects for the injured heart not only by minimizing initial damage of cardiac muscle cells but also by promoting angiogenesis and neovascularization and thus allowing proper perfusion. Currently, we are investigating the signal transduction mechanisms mediating lmwFGF-2 cardioprotection by focusing on cardiac mitochondria as an important target of both indirect (plasma membrane-mediated) and direct lmw-FGF-2 signaling.
In contrast to the beneficial effects of lmwFGF-2, the hmwFGF-2 isoform can exert deleterious effects in the heart, including the induction of pathological growth (hypertrophy) and fibrosis. Many experimental models of heart disease are associated with increased levels of hmwFGF-2. We are examining strategies for neutralizing the detrimental effects of hmwFGF-2 while maintaining the beneficial effects of lmwFGF-2.
(2) Cardiac Remodeling and the Role of Secreted FGF-2 Isoforms
Cardiac fibroblasts represent a major heart cell population, they are responsible for producing connective tissue and proper cardiac architecture. Many chronic cardiac diseases are associated with cardiac fibroblast ‘activation’ to a hyper-secretory version that initially promotes wound healing but eventually creates maladaptive changes including fibrosis and heart failure. We have found that cardiac activated fibroblasts are the major source of cardiac hmwFGF-2, which becomes secreted by these cells and goes on to induce hypertrophy on the muscle cells as well as a pro-fibrotic response by the secreting cells. We are currently using genetically engineered mouse models, expressing only hmwFGF-2 or only lmwFGF-2, to further study the role of these FGF-2 isoforms in the development of chronic heart disease and ability for repair-regeneration.
(3) Translation to Human
We have established that human patient cardiac tissue (atrial tissue fragments, routinely dissected during cardiac vascular surgery) is relatively rich in FGF-2 isoforms, and that fibroblasts grown from human tissue produce biologically active hmwFGF-2, with similar properties as the one from our experimental mouse or rat models. Thsu we have established that our findings from experimental models are applicable to the human system. We are currently investigating mechanisms of regulation of hmwFGF-2 abundance and secretion by human heart cells, and how various drugs (used to treat heart patients) or nutraceuticals such as resveratrol, may affect hmwFGF-2 expression and secretion.
(4) Regulation of and by Connexin-43
We are investigating the role of connexin-43, the main cardiomyocyte gap junction channel protein, in the regulation of cardiomyocyte signaling relating to cell-cell communication, growth, gene expression and mitochondrial-based cardioprotection. This project studies specific post-translational modifications of Cx43 and uses engineered Cx43 mutations and deletions to alter the properties of the molecule and is accompanied by proteomics studies to identify interacting partners of connexin-43 in the normal, injury-protected, or ‘stressed’ state.
Why is this work important?
Our work addresses fundamental issues in regards to the normal and pathological condition of the heart. Induction of cardioprotection and improved heart repair are very attractive approaches aiming at reducing cardiac tissue loss and dysfunction. Our work can identify means by which heart resistance to injury/cell death can be augmented. Our work is also describing (for the first time) the dichotomous functional properties of an endogenous protein and has the potential to identify strategies for reducing the deleterious version and augmenting the beneficial one.
What techniques are used in this laboratory
We are essentially a protein biochemistry and cell biology laboratory. Regular established techniques include cardiac cell isolation and culture, recombinant protein isolation, purification and analysis, protein detection, transient gene transfer, immunofluorescence microscopy (regular and confocal), subcellular fractionation, in vitro and in vivo models of ischemic injury, protein-protein interactions, isolated perfused rat heart approaches. We have standard cell biology – cell culture, histology lab equipment.
About Elissavet Kardami:
E. Kardami obtained her Bachelor’s Degree in Biology, at the University of Athens, Greece (1975), and her PhD degree (1980) from the Department of Cell Biophysics, King’s College, London UK. She pursued post-doctoral studies at the Institute Pasteur, Paris, France and at Berkeley, University of California. She started her independent career as a Faculty member of the University of Manitoba, and a staff scientist at St. Boniface Hospital Research Centre in 1987. She is now a full professor in the Department of Human Anatomy and Cell Sciences, University of Manitoba, and a lab director (Muscle Cell Biochemistry) at the Institute of Cardiovascular Sciences, St. Boniface Research Centre. She has trained 7 PhD, 5 MSc and 4 BSc.Med students, is currently supervising 2 PhD students, two research associates, and one technician, and is funded by the CIHR, and Heart and Stroke Foundation of Manitoba. She has given over 90 invited presentations world-wide, published 87 peer-reviewed papers, and 10 book chapters to-date. In 2009, she received the YMCA Woman of Distinction Award.
For more information, please contact:
Dr. Elissavet Kardami
Email. ekardami@sbrc.ca
In Detail
Dr. Elissavet Kardami is studying how heart muscle and non-muscle cells respond to an injury stimulus, how they repair themselves after injury, and how the potential for subsequent cardiac failure can be prevented. She is tackling these issues by examining the activity of a growth factor protein normally present in the heart and throughout the body in different forms, a beneficial one (lmwFGF) and a detrimental one (hmwFGF). “The growth factors are the molecules the body uses anyway, but they are somewhat suppressed or modified in adults,” Dr. Kardami explains. “If we can subtly change their levels or properties to reactivate them, it may protect the heart’s health without many side effects.”
The Self-Defense Molecule
A potential medical application for one of the growth factor versions Dr. Kardami is studying lies in the ability of lmwFGF to protect cells during or immediately following a myocardial infarction (heart attack). Because the protein appears to make cells and tissues more resistant to injury, the hypothesis is that if it is expressed/administered in greater quantities in the heart, it can protect the organ from certain degrees of injury and improve its maintenance. Ideally, ways would be devised to stimulate the expression of the protein within the heart. However, Dr. Kardami speculates that, as a shortcut, the growth factor can be introduced into the patient’s circulatory system or into the myocardium itself upon arrival in the hospital or even en route to reduce the extent of cell destruction. The lab’s studies have confirmed that this is, in fact, the case in several animal models. Recombinant technology makes production of sufficient quantities of the substance feasible for this application, and further developments surrounding the growth factor’s clinical potential are expected in the near future.
The Heart Failure Molecule
Since the hmwFGF version of the growth factor Dr. Kardami is studying correlates well with cardiac chronic pathologies (hypertrophy, heart failure), its relative levels (in body fluids, tissue, or in vitro systems) can potentially serve as a marker for the efficacy of various current or experimental drug treatments against a heart failure phenotype. Strategies aimed at reducing the relative levels of hmwFGF, while maintaining or increasing those of lmwFGF in vivo would be expected to be beneficial in preventing or delaying transition to heart failure, in the aging or cardiomyopathic heart.
Channeling or Not with Connexins
Another aspect of this laboratory’s studies involves the role of the same growth factors in the way heart cells communicate. In order to act as an efficient pump, the heart’s many muscle cells must contract in simultaneous fashion. This coordination is made possible by channels (termed gap junctions) between cells and the passage of a variety of small molecules (signals) through gap junctions. Abnormalities in gap junction proteins are linked to arrhythmias. The growth factor proteins plays a role in determining the size and function of the channels (made of connexins) and the number and type of molecules that move through them at any time. This indicates that growth factors also have a role in arrhythmias. These studies are currently being conducted at the cell culture and whole heart level.
Selected Recent Publications
Sontag D, Wang J, Kardami E, Cattini PA. FGF-2 and FGF-16 Protect Isolated Perfused Mouse Hearts from Doxorubicin-Induced Contractile Dysfunction. Cardiovasc Toxicol. 2013 Feb 21. [Epub ahead of print]
Lee D, Oka T, Hunter B, Robinson A, Papp S, Nakamura K, Srisakuldee W, Nickel BE, Light PE, Dyck JRB, Lopaschuk GD, Kardami E, Opas M, Michalak M. Calreticulin Induces Dilated Cardiomyopathy. PLoS One. 8(2):e56387, 2013
Wang J, Nachtigal M, Kardami E, Cattini PA. FGF-2 protects cardiomyocytes from doxorubicin damage via protein kinase C-dependent effects on efflux transporters. Cardiovasc Res. 98, 56-63, 2013
Jeyaraman, MM, Srisakuldee W, Nickel BE, Kardami E. The role of connexin43 in cytoprotection. Biochem Biophys Acta-Biomembranes, 1818, 2008-2012, 2012.
Jimenez SK, Jassal DS, Kardami E, Cattini PA. Protection by endogenous FGF-2 against isoproterenol-induced cardiac dysfunction is attenuated by cyclosporine. Mol Cell Biochem. 357, 1-8, 2011
Jimenez SK, Jassal DS, Kardami E, Cattini PA. A Single Bout of Exercise Promotes Sustained Left Ventricular Function Improvement after Isoproterenol-Induced Injury in Mice. J Physiol Sci 61,331-336, 2011
Santiago JJ, Ma X, McNaughton L, Nickel BE, Yu L, Fandrich RR, Netticadan T, Kardami, E. Preferential accumulation and release of high molecular weight FGF-2 by rat cardiac non-myocytes. Cardiov Res 89, 139-47, 2011
Santiago JJ, Dangerfield A, Rattan S, Bathe KL, Cunnington R, Raizman J, Bedosky KM, Freed D, Kardami E, Dixon IMC. Cardiac fibroblast to myofibroblast differentiation in vitro:expression of focal adhesion components in neonatal and adult rat ventricular myofibroblasts. Dev.Dynamics, 239, 1573-84, 2010
Srisakuldee W, Jeyaraman MM, Nickel BE, Tanguy S, Jiang Z-S, Kardami E. Phosphorylation of cardiac connexin-43 at serine 262 promotes an injury-resistant state. Cardiov.Res. 83, 672-81, 2009
Kardami E, Detillieux K, Ma X, Jiang ZS, Santiago JJ, Jimenez SK, Cattini PA. FGF-2 and cardioprotection. Heart Fail Rev. 2007 Dec;12(3-4):267-77.
Zhang Y, Kanter EM, Laing JG, Kardami E, Yamada K. Connexin43 expression levels influence intercellular coupling and cell proliferation of native murine cardiac fibroblasts. Cell Commun Adhes. 2008, 15, 289-303
Ma X, Dang X, Claus P, Hirst, Fandrich RR, Jin Y, Grothe C, Kirshenbaum L, Cattini PA and Kardami E. Chromatin compaction and cell death by high molecular weight FGF-2 depend on its nuclear localization, intracrine activation of ERK1/2 and engagement of mitochondria. Journal of Cellular Physiology 213, 690-698, 2007.
Jiang ZS, Jeyaraman M, Wen G-B, Fandrich RR, Dixon IAC, Cattini PA and Kardami E. High- but not low-molecular weight FGF-2 causes cardiac hypertrophy in vivo: Possible involvement of cardiotrophin-1. J Mol Cell Cardiol 42:222-233, 2007.
Kardami E, Dang X, Iacobas DA, Nickel BE, Jeyaraman M, Srisakuldee W, Makazan J, Tanguy S, Spray DC. The role of connexins in controlling cell growth and gene expression Progress in Biophysics and Molecular Biology, 94, 245-264, 2007.
Selected Peer-Reviewed Publications
Kardami E and Fandrich RR. Basic fibroblast growth factor in atria and ventricles of the vertebrate heart. J Cell Biology 109:1865-1875, 1989
Pasumarthi SKB, Kardami E and Cattini PA. High and low molecular weight FGF-2 increase proliferation of neonatal rat cardiac myocytes but have differential effects on binucleation and nuclear morphology. Circ Res 78:126-136, 1996
Doble BW, Bosc D, Litchfield D and Kardami E. Fibroblast growth factor-2 decreases metabolic coupling and stimulates phosphorylation as well as masking of connexin43 epitopes in cardiac myocytes. Circ Res 79:647-658, 1996
Padua RR, Merle P-L, Doble BW, Yu C-H, Zahradka P, Pierce GN, Panagia V and Kardami E. Fibroblast growth factor-2-induced negative inotropism and cardioprotection are inhibited by chelerythrine; involvement of sarcolemmal calcium-independent protein kinase C. J Mol Cell Cardiol 30:2695-2709, 1998
Sheikh F, Fandrich RR, Kardami E and Cattini PA. Overexpression of long and short FGFR-1 isoforms results in FGF-2-mediated proliferation in neonatal cardiac myocyte cultures. Cardiovasc Res 42:696-705, 1999
Doble BW, Ping P and Kardami E. The epsilon subtype of protein kinase C is required for cardiomyocyte connexin-43 phosphorylation. Circ Res 86:293-301, 2000
Sheikh F, Sontag DP, Kardami E and Cattini PA. Exogenous addition and endogenous overexpression of FGF-2 increases cardiac myocyte viability after myocardial injury in isolated mouse hearts. Am J Physiol 280:H1039-H1050, 2001
Sun G-P, Doble BW, Sun J-M, Florkiewicz F, Kirshenbaum L, Davie JR, Cattini PA and Kardami E. CUG-initiated FGF-2 induces chromatin compaction in cultured cardiac myocytes and in vitro. J Cell Physiol 186:457-467, 2001
Jiang Z-S, Padua RR, Ju H, Doble BW, Jin Y, Hao-J, Cattini PA, Dixon IMC and Kardami E. Acute protection of the ischemic heart by FGF-2; involvement of FGF-2 receptors and protein kinase C. Am J Physiol 282:H1071-H1080, 2002
Dang X, Doble BW and Kardami E. The carboxy-tail of connexin-43 localizes to the nucleus and inhibits cell growth. Mol Cell Biochem 242, 35-38, 2003
Jeyaraman M, Tanguy S, Fandrich RR, Lukas A and Kardami E. Ischemia-induced dephosphorylation of cardiomyocyte connexin-43 is reduced by okadaic acid and caliculin A but not fostriecin. Mol Cell Biochem 242:129-134, 2003
Doble BW, Dang X, Ping P, Fandrich RR, Nickel BE, Jin Y, Cattini PA and Kardami E. Phosphorylation of serine 262 in the gap junction protein connexin-43 regulates DNA synthesis in cell-cell contact forming cardiomyocytes. J Cell Sci 2004 117:507-514, 2004
Jiang ZS, Srisakuldee W, Soulet F, Bouche G and Kardami E. Non-angiogenic FGF-2 protects the ischemic heart from injury, in the presence or absence of reperfusion. Cardiovasc Res 62:154-166, 2004
Jimenez SK, Sheikh F, Jin Y, Detillieux KA, Dhaliwal J, Kardami E and Cattini PA. Transcriptional regulation of FGF-2 gene expression in cardiac myocytes. Cardiovasc Res 62, 548-557, 2004
Kardami E, Jiang ZS, Jimenez SK, Hirst CJ, Sheikh F, Zahradka P and Cattini PA. The role of FGF-2 isoforms in cardiac hypertrophy. Cardiovasc Res 63:458-466, 2004
Jiang ZS, Jeyaraman M, Wen G-B, Fandrich RR, Dixon IAC, Cattini PA and Kardami E. High- but not low-molecular weight FGF-2 causes cardiac hypertrophy in vivo: Possible involvement of cardiotrophin-1. J Mol Cell Cardiol 42:222-233, 2007.
Kardami E, Dang X, Iacobas DA, Nickel BE, Jeyaraman M, Srisakuldee W, Makazan J, Tanguy S, Spray DC. The role of connexins in controlling cell growth and gene expression Progress in Biophysics and Molecular Biology, 94, 245-264, 2007 .
Ma X, Dang X, Claus P, Hirst, Fandrich RR, Jin Y, Grothe C, Kirshenbaum L, Cattini PA and Kardami E. Chromatin compaction and cell death by high molecular weight FGF-2 depend on its nuclear localization, intracrine activation of ERK1/2 and engagement of mitochondria. Journal of Cellular Physiology 213, 690-698, 2007.
Kardami E, Detillieux K, Ma X, Jiang ZS, Santiago JJ, Jimenez SK, Cattini PA. FGF-2 and cardioprotection. Heart Fail Rev. 2007 Dec;12(3-4):267-77.
Srisakuldee W, Jeyaraman MM, Nickel BE, Tanguy S, Jiang Z-S, Kardami E. Phosphorylation of cardiac connexin-43 at serine 262 promotes an injury-resistant state. Cardiov.Res. 83, 672-81, 2009
Santiago JJ, Dangerfield A, Rattan S, Bathe KL, Cunnington R, Raizman J, Bedosky KM, Freed D, Kardami E, Dixon IMC. Cardiac fibroblast to myofibroblast differentiation in vitro:expression of focal adhesion components in neonatal and adult rat ventricular myofibroblasts. Dev.Dynamics, 239, 1573-84, 2010
Santiago JJ, Ma X, McNaughton L, Nickel BE, Yu L, Fandrich RR, Netticadan T, Kardami, E. Preferential accumulation and release of high molecular weight FGF-2 by rat cardiac non-myocytes. Cardiov Res 89, 139-47, 2011
Jimenez SK, Jassal DS, Kardami E, Cattini PA. A Single Bout of Exercise Promotes Sustained Left Ventricular Function Improvement after Isoproterenol-Induced Injury in Mice. J Physiol Sci 61,331-336, 2011
Jimenez SK, Jassal DS, Kardami E, Cattini PA. Protection by endogenous FGF-2 against isoproterenol-induced cardiac dysfunction is attenuated by cyclosporine. Mol Cell Biochem. 357, 1-8, 2011
Jeyaraman, MM, Srisakuldee W, Nickel BE, Kardami E. The role of connexin43 in cytoprotection. BBA-Biomembranes, 2011. [Epub ahead of print]
1) the Canadian Institutes for Health Research (2010-1015)
2) the Heart and Stroke Foundation of Manitoba (2012-2014)
Life In the Lab
