RESEARCH PROGRAM

John W. Mellors, M.D.Professor of Medicine

Chief, Division of Infectious DiseasesDirector, HIV/AIDS Program, UPMC

mellors@dom.pitt.eduwww.pitt.edu/~disease

Basic Research

Translational Research

Clinical Research

Better Treatment and Prevention

Understand Drug ResistanceDiscover New Drugs

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

Doctoral Student Training Model

Important Clinically Relevant Question?

Genetic, Virologic, Biochemical,

Structural Studies

C. Shi, PhD1991-1997

Development of recombinant retroviral systems for analysis of HIV-1 drug resistance

J. Hammond, PhD1996-2002

Structural determinants of HIV resistance to nucleoside analogs

U. Parikh, PhD1999-2005

Antagonistic evolutionary pathways of HIV-1 resistance to NRTI

E. Halvas, PhD2000-Present

Role of low frequency drug-resistant variants in antiretroviral treatment failure

S. Clark, PhD2002-Present

Genetic, biochemical, and structural mechanisms of NNRTI hypersusceptibility

J. Brehm2005-Present

The influence of mutations in the polymerase and RNase H domains on resistance to reverse transcriptase inhibitors

J. Meeter2007-

PresentStructural basis of nucleoside activity, resistance and toxicity

Past/Current Doctoral Trainees

Jessica BrehmPre-doctoral Student

IDM Department - GSPHAnd member of

Mellors Lab since 2004

BS Biochemistry and Chemistry 2003College of Charleston

South Carolina

PEER-REVIEWED PUBLICATION

1. Brehm JH, Koontz D, Meteer JD, Pithak V, Sluis-Cremer N, Mellors JW. Selection of Mutationsin 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)2. XVI International HIV Drug Resistance Workshop June 2007, Barbados.3. 15th Annual CROI conference, February 2008, Boston, MA (Oral presentation)4. XVII International HIV Drug Resistance Workshop, June 2008, Sitges, Spain (Oral)

Fellowship: Pittsburgh AIDS Research TrainingFellow (PART) – 2007-08

Jeff MeteerPre-doctoral Student

Molecular Virology and Microbiology Graduate Program MVM

BS Biochemistry 2006St. John Fisher College,

Rochester NY

Member of

Mellors Lab since 2007

PEER-REVIEWED PUBLICATION

Brehm JH, Koontz D, Meteer JD, Pithak V, Sluis-Cremer N, Mellors JW. Selection ofMutations in the connection and RNase Hdomains of human immunodeficiency virusType 1 reverse transcriptase thatIncrease 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 potentNucleoside analogue reverse transcriptase Inhibitor 3’-azido-2’,3’-dideoxyguanosine. ORAL PRESENTATION at the XVII InternationalHIV Drug Resistance Workshop. June 2008Sitges, Spain.

Urvi Parikh, PhDUniversity of Pittsburgh

Assistant Professor of MedicineAssociate Director

Microbial Trials Network Lab

Doctoral student inMELLORS LAB

1999-2005PhD – IDM GSPH 2006BS Biochemistry and

Molecular Biology - 1998Pennsylvania State University

Urvi’s Carrer Pathafter Mellors Lab….(5 publications, 4 first author)

2006 to 2008Emerging Infectious Disease Research Fellow

Centers for Disease Control and PreventionLaboratory of Walid Heneine, PhD, Division of

HIV/AIDS Prevention Laboratory Branch.

Conducted in vitro evaluations of nucleoside reversetranscriptase 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 2008To serve as Associate Director of Microbial TrialsNetwork and to enhance ID Division global health

Initiatives

Nicolas Sluis-Cremer, PhDUniversity of Pittsburgh

Assistant Professor

Director, Basic EducationID Division

Jessica RadzioPre-doctoral Student

IDM Department of GSPHSluis-Cremer Lab since 2007

BS Biology - 2001MS Life Sciences - 2003

Virginia Polytechnic & StateUniversity, Blacksburg, VA

PEER-REVIEWED PUBLICATIONS 2007-081. Radzio J, Sluis-Cremer N. Efavirenz acceleratesHIV-1 reverse transcriptase ribonuclease H cleavage, leading to diminished zidovudine excision. Mol Pharmacol 2008;73(2)601-6.

2. Zelina S, Sheen CW, Radzio J, Mellors JW andSluis-Cremer N. Mechanisms by which the G333DMutation in human immunodeficiency virus type 1Reverse transcriptase facilitates dual resistanceProtein Sci 2007;16(8) 1728-37.

3. Xia Q, Radzio J and Sluis-Cremer N. Probing Non-nucleoside inhibitor-induced active-site distortion in HIV-1 reverse transcriptase bytransient 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 distortionIn HIV-1 reverse transcriptase by RT. Biochem J2007;404(1) 151-7.

Brian HermanPre-doctoral Student

Molecular Virology andMicrobiology Graduate Program

BS Biology/BiochemistryAllegheny College 2005

Sluis-Cremer Lab2007

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. AMouse-adapted SARS-coronavirus causes diseaseand mortality in BALB/cmice. PLoS Pathog 2007;3(1):e5 T.

PhD Thesis: Structure-activity-resistance Relationships and biochemicalPharmacology of novel nucleoside andNucleotide HIV-1 reverse transcriptaseinhibitors

S. Riddler, MD 1991-1994 Antiretroviral activity of stavudine (2’,3’-didehydro’3’-depxythymiding, d4T)

H. Nguyen, MD 1991-1995Molecular basis of HIV resistance to NNRTI

N. Connolly, MD 2002-2005 Immune based therapies for HIV infection

J. McKinnon, MD 2003-2005Simplified maintenance antiretroviral therapy: Virologic predictors of outcome

S. Patel, DO 2003-2005Persistence of multidrug resistant HIV after structured treatment interruptions

J. Chow, BS 2007-PresentRole of minor drug-resistant variants in treatment failure

Past MD Fellow Trainees

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

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

Mellors LabUniversity of Pittsburgh

Division Infections Diseases