RESEARCH PROGRAM

1. RESEARCH PROGRAM John W. Mellors, M.D. Professor of Medicine Chief, Division of Infectious Diseases Director, HIV/AIDS Program, UPMC mellors@dom.pitt.edu www.pitt.edu/~disease

2. Basic Research Translational Research Clinical Research Understand Drug Resistance Discover New Drugs Better Treatment and Prevention

3. Major Projects • Discovery of novel nucleoside analogs with activity against drug-resistant HIV • New mechanisms of HIV drug resistance • Role of low frequency drug-resistant variants in treatment failure • Mechanisms of HIV persistence on antiretroviral therapy and strategies for eradication • HIV prevention with antiretrovirals

4. Doctoral Student Training Model Important Clinically Relevant Question? Genetic, Virologic, Biochemical, Structural Studies

5. Past/Current Doctoral Trainees

6. PEER-REVIEWED PUBLICATION • Brehm JH, Koontz D, Meteer JD, Pithak V, • Sluis-Cremer N, Mellors JW. Selection of Mutations • in the Connection and RNase H Domains of Human • Immunodeficiency Virus Type 1 Reverse • Transcriptase that Increase Resistance to 3'-Azido- • 3'-Dideoxythymidine. J Virol 2007;81:7852-9 • ABSTRACTS Presented at Conferences 2007-08 • 1. 14th Annual Conference on Retroviruses and • Opportunistic Infections (CROI). Feb 2007, • Los Angeles, CA. (Oral presentation) • XVI International HIV Drug Resistance • Workshop June 2007, Barbados. • 15th Annual CROI conference, February 2008, • Boston, MA (Oral presentation) • XVII International HIV Drug Resistance • Workshop, June 2008, Sitges, Spain (Oral) • Fellowship: Pittsburgh AIDS Research Training • Fellow (PART) – 2007-08 Jessica Brehm Pre-doctoral Student IDM Department - GSPH And member of Mellors Lab since 2004 BS Biochemistry and Chemistry 2003 College of Charleston South Carolina

7. PEER-REVIEWED PUBLICATION Brehm JH, Koontz D, Meteer JD, Pithak V, Sluis-Cremer N, Mellors JW. Selection of Mutations in the connection and RNase H domains of human immunodeficiency virus Type 1 reverse transcriptase that Increase resistance to 3’-azido-3’- Dideoxythymidine. J Virol 2007;81: 7852-9. ABSTRACT Presented at Conference: Meteer J, Koontz D, Rapp KL, Detorio M, Ruckstuhl M, Schinazi RF, Mellors JW. Novel resistance profile of the potent Nucleoside analogue reverse transcriptase Inhibitor 3’-azido-2’,3’-dideoxyguanosine. ORAL PRESENTATION at the XVII International HIV Drug Resistance Workshop. June 2008 Sitges, Spain. Jeff Meteer Pre-doctoral Student Molecular Virology and Microbiology Graduate Program MVM BS Biochemistry 2006 St. John Fisher College, Rochester NY Member of Mellors Lab since 2007

8. Urvi’s Carrer Path after Mellors Lab….(5 publications, 4 first author) 2006 to 2008 Emerging Infectious Disease Research Fellow Centers for Disease Control and Prevention Laboratory of Walid Heneine, PhD, Division of HIV/AIDS Prevention Laboratory Branch. Conducted in vitro evaluations of nucleoside reverse transcriptase inhibitors for use as a microbicide gel for the prevention of HIV transmission. Also involved In design and implementation of candidate Microbicide safety and efficacy studies in non-human primates. RECRUITED TO PITT FACULTY IN 2008 To serve as Associate Director of Microbial Trials Network and to enhance ID Division global health Initiatives Urvi Parikh, PhD University of Pittsburgh Assistant Professor of Medicine Associate Director Microbial Trials Network Lab Doctoral student in MELLORS LAB 1999-2005 PhD – IDM GSPH 2006 BS Biochemistry and Molecular Biology - 1998 Pennsylvania State University

9. Nicolas Sluis-Cremer, PhD University of Pittsburgh Assistant Professor Director, Basic Education ID Division

10. PEER-REVIEWED PUBLICATIONS 2007-08 • 1. Radzio J, Sluis-Cremer N. Efavirenz accelerates • HIV-1 reverse transcriptase ribonuclease H • cleavage, leading to diminished zidovudine • excision. Mol Pharmacol 2008;73(2)601-6. • Zelina S, Sheen CW, Radzio J, Mellors JW and • Sluis-Cremer N. Mechanisms by which the G333D • Mutation in human immunodeficiency virus type 1 • Reverse transcriptase facilitates dual resistance • Protein Sci 2007;16(8) 1728-37. • Xia Q, Radzio J and Sluis-Cremer N. Probing • Non-nucleoside inhibitor-induced active-site • distortion in HIV-1 reverse transcriptase by • transient kinetic analyses. Protein Sci 2007(16) • (8);1728-37 • 4. Nissley DV, Radzio J, Anderson KS, Sluis- • Cremer N. Characterization of novel non- • Nucleoside inhibitor-induced active-site distortion • In HIV-1 reverse transcriptase by RT. Biochem J • 2007;404(1) 151-7. Jessica Radzio Pre-doctoral Student IDM Department of GSPH Sluis-Cremer Lab since 2007 BS Biology - 2001 MS Life Sciences - 2003 Virginia Polytechnic & State University, Blacksburg, VA

11. PEER REVIEWED PUBLICATION: Roberts A, Deming D, Paddock CD, Cheng A, Yount B, Vogel L, Herman BD, Sheahan T, Heise M, Genrich GL, Zaki SR, Baric R, Subbarao K. A Mouse-adapted SARS-coronavirus causes disease and mortality in BALB/cmice. PLoS Pathog 2007;3(1):e5 T. PhD Thesis: Structure-activity-resistance Relationships and biochemical Pharmacology of novel nucleoside and Nucleotide HIV-1 reverse transcriptase inhibitors Brian Herman Pre-doctoral Student Molecular Virology and Microbiology Graduate Program BS Biology/Biochemistry Allegheny College 2005 Sluis-Cremer Lab 2007

12. Past MD Fellow Trainees

13. Laboratory Techniques • Experimental design! • Cell culture and virology • Antiviral susceptibility/cytotoxicity assays • drug assessments • Biochemistry of RT and other enzymes • Molecular biology • Cloning, PCR, site-specific mutagenesis, DNA sequencing, single genome sequencing • Structural biology • Application of above to human samples

14. Expectations • Hard work and commitment to science • Weekly meetings with supervisor • 2 or 3 first author publications (for Ph.D) • Scientific presentations at national meetings

15. Mellors Lab University of Pittsburgh Division Infections Diseases