David Graham, PhD

Assistant Professor, Molecular and Comparative Pathobiology
and joint appointment Department of Medicine, Division of Cardiology

Background & Training:

Dr. Graham relocated from Canada following his undergraduate studies at the University of Guelph and Masters studies at McMaster University, where he studied several viral pathogens in the context of mucosal immunology, to work and eventually help to manage a group in the AIDS Vaccine Program at NCI-Frederick in 1997.  After a two-year break from academics he returned to graduate school at Johns Hopkins, where he obtained his Ph.D from the Biochemistry, Cellular and Molecular Biology Training program in the Department of Pharmacology and Molecular Sciences under the direction of Dr. James Hildreth, where he studied the role of virion-associated lipid rafts.  After his Ph.D., Dr. Graham joined the faculty as a research associate in the laboratory of Dr. Jennifer Van Eyk in Department of Medicine, Division of Cardiology at Bayview where he obtained training in proteomics and applied these skills to several cardiology related areas, including the study of a transgenic rabbit model of human hypertropic cardiomyopathy and the function of sino-atrial nodal cells.   Dr. Graham used this opportunity to apply his acquired skills in proteomics to the emerging field of viral proteomics, where he has developed some of the first proteomics methods to study viruses and post-translational modifications of viral and viral associated host proteins.

Research:

Dr. Graham’s research interests in retroviruses came from his early observations that in the Human and Simian Immunodeficiency viruses, virions acquired several host proteins involved in antigen processing and presentation, and that viruses more capable of packaging these host-proteins exhibited more prominent pathology in vitro.  This led to him asking whether these host proteins were functionally organized into membrane microdomains in budded virus, similar to how the proteins would be organized in the cell.  Using a variety of methods he was able to demonstrate that virions do contain organized lipid raft domains, and that these domains could be targeted to irreversibly inactivate both HIV and SIV using the compound 2 hydroxyl-beta-cyclodextrin.  This compound is now under study as an anti-HIV microbicide by the Hildreth laboratory.  The complex systems that were now under study by Dr. Graham required the development of completely new methods in virology due to the low abundance of these proteins and the incredible diversity of host-proteins packaged into retroviruses.  Coupling mass spectrometry to existing methods of protein separation along with the development of software workflows to analyze the complex data, Dr. Graham has been able to identify over 160 host-proteins from virion-associated lipid rafts and methods to determine the site-specific post-translational modifications of viral proteins.  His current research is concentrated on understanding the differences between viruses with divergent pathologies, including the ability to differentially induce apoptosis by a IFN a/B induced expression of TRAIL, in collaboration with Gene Shearer (NIH) and Jean-Phillipe Herbeuval (Necker Institute, France), and viruses with the ability to induce neurotoxicity in collaboration with Drs. Zink and Clements (retrovirus laboratory).

Dr. Graham’s research program in Cardiology includes the molecular phenotyping of transgenic rabbits over expressing alpha-myosin heavy chain, which has been shown to be cardioprotective against pacing induced heart failure in collaboration with Dr. Jeffrey Robbins (Children’s institute, Cincinnatti).  In this complex experimental model, Dr. Graham has been able to elucidate changes in the proteome caused by the transgene alone as compared to the background strain and differences between all groups under pacing induced heart failure.  In collaboration with Dr. Edward Lakatta (NIA, GRC), Dr. Graham has been investigating the role of lipid rafts in the control of SA nodal cell function.  He has developed ultra-sensitive mass-spectrometry methods to be able to compare compositional changes in the lipid raft proteome between control and stimulated SA nodal cells.  Dr. Graham in collaboration with Dr. Joe Mankowski (Retrovirus Laboratory) hopes to merge his research programs in cardiology and virology to be able to study HIV-induced cardiomyopathy, an important emerging area in Cardiovascular Medicine.

Committees / Panels:

American Heart Association - Committee: Basic Cell and Molecular Biology 2,    National Committee, Reviewer, Appointed from March 2006 to March 2010

Maryland Industrial Partnerships (MIPS), Reviewer

Department of Health & Human Services – Cardiovascular Sciences Small Business Activities Review Committee, Reviewer

Relevant recent articles:

1.      David R.M. Graham.  HIV and other Viral Screens (Van Eyk, Dunn (Eds)):  Clinical Proteomics. From Diagnosis to Therapy, In Press, April 2007.

2.      Herbeuval JP and Shearer GM, HIV-1 Immunopathogenesis: How good interferon turns bad. Clin Immunol. 2006 Nov 15

3.      David R. M. Graham*, Megan Mitsak, Steven T. Elliott, Dawn Chen, Steven A. Whelan, Gerald W. Hart and Jennifer E. Van Eyk.  Determination of Site-Specific Glycosylation Usage in Human and Simian Immunodeficiency Virus:  Towards Clinical Assessment of Viral Glycosylation. Molecular and Cellular Proteomics (submitted).

4.      Yurong Guo, Patrick A. Singleton, Austin Rowshan, Marjan Gucek, Robert N. Cole, David R. M. Graham, Jennifer E. Van Eyk and Joe G.N. Garcia, Quantitative Proteomic Analysis of Endothelial Cell Lipid Rafts: Evidence of MARCKS and MRP Regulation in the Sphingosine 1-phosphate Induced Barrier Enhancement, Molecular and Cellular Proteomics (2007, January 8, Epub ahead of print)

5.      David R. M. Graham, Steven T. Elliott and Jennifer E. Van Eyk, Broad-Based Proteomic Strategies:  A Practical Guide to Proteomics and Disease Screening.  J Physiol. 2004 Dec 20.

6.      David R. M. Graham, Christopher P. Garnham, Qin Fu, Jeffrey Robbins  and Jennifer E. Van Eyk, Improvements in Two-Dimensional Gel Electrophoresis by Utilizing a Low Cost “In-House” Neutral pH SDS-PAGE Electrophoresis System.  Proteomics (Proteomics 2005 Jun;5(9):2309-14).

7.      Malini Mansharamani, David R. M. Graham, Daphne Monie, Kenneth K. Lee, James E.K. Hildreth, Robert F. Siliciano and Katherine L. Wilson,  Barrier-to-autointegration Factor Binds Gag and is a Stable Component of HIV-1 Virions. J Virol. 2003 Dec; 77(24): 13084-92.

8.      David R. M. Graham, Elena Chertova, Joanne Hilburn, Larry Arthur and James E.K. Hildreth, Cholesterol depletion of HIV-1 and SIV with Beta-cyclodextrin inactivates and permeabilizes the virions: Evidence for virion-associated lipid rafts. J Virol. 2003 Aug; 77(15): 8237-48.

9.      Zhaohao Liao, David R. M. Graham, and James E.K. Hildreth.  Lipid rafts and HIV Pathogenesis: Cholesterol is required for Fusion and Infection of Susceptible Cells.  AIDS Research and Human Retroviruses, 2003 Aug; 19(8): 675-87.

10.  Mark T. Esser, David R. M. Graham, Lori V. Coren, Charles M. Trubey,        Julian W. Bess, Jr., Larry O. Arthur, David E. Ott and Jeffrey D. Lifson. Differential Incorporation of CD45, CD80 (B7-1), CD86 (B7- 2), MHC-I and MHC-II into HIV-1 Virions and Microvesicles: Implications for Viral Pathogenesis and Immune Regulation. J Virol. 2001 Jul;75(13):6173-82.