Anthony Gramolini

Research Synopsis

Keywords: Ca2+ cycling, cardiac muscle, dilated cardiomyopathy, cell biology, proteomics, biomarkers, heart failure.

Detailed Description:
A major interest of our lab is to investigate the cellular mechanisms involved in the regulation of disease progression in heart failure. We have also had a longstanding interest in Ca2+ cycling at the level of sarcoplasmic reticulum and its role in cardiac disease. For these experiments, we apply conventional biochemical and molecular biology assays, digital confocal imaging, and mass spectrometry based proteomic studies. Model systems include: tissue explants, established cell lines, primary cell cultures and transgenic animals. These techniques serve as valuable experimental tools to investigate heart disease pathways, cellular protein sorting and Ca2+ cycling.

METHODS USED

Cell and tissue culture: Cardiomyocytes, human biopsies, primary cultured cells and ES-derived stem cells.

Procedures: Cell biology, confocal imaging, HPLC, gene expression analysis, immunohistochemistry, mass spectrometry, mircoarrays, proteomics, qRT-PCR, RT-PCR, siRNA, transgenic mice, western blot.

EQUIPMENT USED

Calcium imaging system (Olympus), confocal microscope (Leica LSM IRBE), culture hood, culture incubators, digital microscope (Zeiss structured illumination (SIM), fluorescence microscope, HPLC (Akta), Kodak documentation system, low- and high-speed centrifuge, mass spectrometer (LTQ Orbitrap, TSQ Vantage), plate reader (Perkin Elmer), setups for electropherosis.

PRESENT TRAINEES

Michelle Di Paola, PhD candidate
Chris Oldfield, PhD candidate
Malak Elbatarny, PhD candidate
Dr. Allen Teng, Research Associate
Dr. Uros Kuzmanov, Research Associate
Dr. Meghan McFadden, Research Associate
Dr. Marjan Tavassoli, PDF
Dr. Cristine Reitz, PDF
Wenping Li, Tech II

PRESENT COLLABORATIONS

Within the Department of Physiology:
Peter Backx

Outside the Department of Physiology:
Gordon Keller, UHN
Peter Liu, UHN
Thomas Kislinger, UHN
Mansoor Husain, UHN
Ian Scott, Sick Kids
Seema Mital, Sick Kids
Robert Hamilton, Sick Kids
Jack Greenblatt, CCBR
Andrew Emili, CCBR
Gary Bader, CCBR
Jason Moffat, CCBR
Igor Stagljar, CCBR
Benoit Bruneau, USCF Gladstone

Recent Publications

1. Lee SH, Hadipour-Lakmehsari S, Kim DH, Di Paola M, Kuzmanov U, Shah S, Lee JJ, Kislinger T, Sharma P, Oudit GY, Gramolini AO. (2020) Bioinformatic analysis of membrane and associated proteins in murine cardiomyocytes and human myocardium. Scientific Data. 2020 Dec 1;7(1):425. doi: 10.1038/s41597-020-00762-1. (*Senior corresponding author)
2. Kuzmanov U, EY Wang, R Vanderlaan, H Guo, S Hadipour-Lakmehsari, Y Zhao, DH Kim, P Sharma, F Billia, M Radisic, A Gramolini*, A Emili* (2020) Integrative Phosphoproteomic Profiling of Clinical Samples, Animal and Organ-On-A-Chip Models: Mapping Cardiac Fibrosis. Nature Biomedical Engineering doi:10.1038/s41551-020-0585-y (*Senior corresponding author)
3. Lee S-H, S Hadipour-Lakmehsari, N Gibb, A.C.T. Teng, T Miyake, J Cosme, M Moon, V Wong, J C. Yu, P Liu, F Billia, R Fernandez-Gonzalez, I Stagljar, P Sharma, T Kislinger, I. C. Scott, A.O. Gramolini. (2020). REEP5 depletion causes endoplasmic reticulum dysfunction and cardiac developmental and functional defects. Nature Communications. 11(1):965. doi: 10.1038/s41467-019-14143-9. (Senior corresponding author)
4. Hadipour-Lakmehsari S, Driouchi A, Lee SH, Kuzmanov U, Callaghan NI, Heximer SP, Simmons CA, Yip CM, Gramolini AO (2019) Nanoscale reorganization of sarcoplasmic reticulum in pressure-overload cardiac hypertrophy visualized by dSTORM. Scientific Reports. 9(1):7867. doi: 10.1038/s41598-019-44331-y. (Senior corresponding author)
5. Kuzmanov U, Guo H, Buchsbaum D, Cosme J, Abbasi C, Isserlin R, Sharma P, Gramolini AO*, Emili A*. (2016) Global phosphoproteomic profiling reveals perturbed signaling in a mouse model of dilated cardiomyopathy. Proc Natl Acad Sci (USA). 13(44):12592-12597. (Senior co-corresponding author).
6. Sharma P, Abbasi C, Bousette N, Lazic S, Dubois N, Ignatchenko A, Ignatchenko V, Teng A, Wilson A, Noronha M, Wong V, Liu J, Araki T, Liu J, Tiburcy M, Zimmerman WH, Ackerley C, Hamilton R, Sun Y, Liu P, Backx P, Keller G, Stagljar I, Scott I, Kislinger T, Gramolini AO. (2015). Tmem65 is an Evolutionary Conserved Membrane Protein that Regulates Cardiac GAP Junction communication. Nature Communications 25, 8391. (Senior corresponding author).

Courses

Course Number: PSL498 (Coordinator), PSL462/1462, PSL1040, PSL1067, JCV3060, JCV3062, JCV3065