focus on CCR5 knockout strategies using Zinc-finger nucleases

1. HIV cure and eradication Rationales and Options Pana Tanjararak, MD

19. Sharon R Lewin and Christine Rouzioux. AIDS 2011; 25: 000-000. Infectious diseases unit, The Alfred, Monash University Melbourne, Australia. Laboratory of virology, Hopital Necker, Universite Paris Descartes, Paris, France.

20. Sharon R Lewin and Christine Rouzioux. AIDS 2011; 25: 000-000. Infectious diseases unit, The Alfred, Monash University Melbourne, Australia. Laboratory of virology, Hopital Necker, Universite Paris Descartes, Paris, France.

42. Sharon R Lewin and Christine Rouzioux. AIDS 2011; 25: 000-000. Infectious diseases unit, The Alfred, Monash University Melbourne, Australia. Laboratory of virology, Hopital Necker, Universite Paris Descartes, Paris, France.

60. Sharon R Lewin and Christine Rouzioux. AIDS 2011; 25: 000-000. Infectious diseases unit, The Alfred, Monash University Melbourne, Australia. Laboratory of virology, Hopital Necker, Universite Paris Descartes, Paris, France.

62. HIV Cure and Eradication Chemokine Receptor 5 Knockout Strategies Using Zinc-finger Nuclease Rongpong Plongla, MD

64. The Life Cycle of Human Immunodeficiency Virus Type 1 Marvin S Reitz Jr and Robert C Gallo. In Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases, 7th ed. 2009 Joan stephenson. JAMA. 2002;287(13):1635-1637 .

65. Mechanism of HIV entry Jose’ A Este’ , Amalio Telenti. Lancet 2007; 370: 81–88 Retrovirology Laboratory IrsiCaixa, Hospital Universitari Germans Trias i Pujol, Universitat Autnoma deBarcelona, Badalona, Spain (J A Este PhD) and Institute of Microbiology, University Hospital Centre, University of Lausanne, Lausanne, Switzerland (A Telenti MD)

68. HIV Tropism and Disease Progression Kuhmann SE, et al. J Viral Entry. 2005;1:4-16. Moore JP, et al. AIDS Res Hum Retroviruses. 2004;20:111-126. Time Since Seroconversion HIV-1 RNA Level Current assays’ X4 detection limit Weeks Years R5 virus X4 virus R5 Infection R5 + X4 Infection

70. CCR5 Wild Type and CCR5  32 CCR5 wild type   CCR5  32 2 normal copies 1 copy of  32 2 copies of  32 wt/wt wt/  32  32 /  32 Standard disease progression Delayed disease progression “ Resistant” to HIV infection Normal Heterozygous Homozygous Liu R, et al. Cell. 1996;86:367-367. Samson M, et al. Nature. 1996;382:722-725. Dean M, et al. Science. 1996;273:1856-1862. Huang Y, et al. Nat Med.1996;2:1240-1243. Michael NL, et al. Nat Med. 1997;3:1160-1162. Eugen-Olsen J, et al. AIDS. 1997;11:305-310.

74. J Allergy Clin Immunol 2008; 122: 22-8 • Latently infected resting memory CD4+ T cells are the best characterized latent reservoir for HIV-1. • Less than 1 cell per 1,000,000 resting CD4+ T cells from patients on HAART harbor latent HIV-1 provirus. • Sequence of latent proviruses does not evolve, which suggests no ongoing viral replication. • Discontinuation of HAART allows viral relapse from latent reservoir. • Patients successfully treated with HAART for longer than 10 years exhibit no appreciable decrease in the size of the latent reservoir. • The persistence of latently infected memory CD4+ T lymphocytes precludes their elimination by HAART alone for the lifetime of the patient. • Other drug-insensitive reservoirs, including brain, macrophages, and hematopoietic stem cells, may also exist. • Latency is likely established and maintained by numerous blocks at multiple steps in the HIV-1 replicative pathway, which potentially complicates eradication strategies. Douglas D. Richman.* David M. Margolis, Martin Delaney, Warner C. Greene, Daria Hazuda, Roger J. Pomerantz. Science 6 March 2009: Vol. 323 no. 5919 pp. 1304-1307 . University of California San Diego

75. Can We Do Better Than HAART?

82. Paula Cannon, CROI2011

86. Flavobacterium okeanokoites

87. Double-strand break

88. Fyodor D. Urnov 1 , Edward J. Rebar 1 , Michael C. Holmes 1 , H. Steve Zhang 1 & Philip D. Gregory Nature Reviews Genetics 11, 636-646 (September 2010)

89. Paula Cannon and Carl June. Current Opinion in HIV and AIDS 2011; 6: 74-79. Molecular Microbiology & Immunology, University ofSouthern California Keck School of Medicine, Los Angeles, California and Department of Pathology andLaboratory Medicine, University of PennsylvaniaSchool of Medicine, Philadelphia, Pennsylvania, USA

90. Paula Cannon, CROI2011

91. Pre-clinical testing using humanized mice

93. Nat Biotechnol. 2010 Aug;28(8):839-47.

94. Nat Biotechnol. 2010 Aug;28(8):839-47.

95. Paula Cannon, CROI2011

100. Paula Cannon, CROI2011

103. Paula Cannon and Carl June. Current Opinion in HIV and AIDS 2011; 6: 74-79. Molecular Microbiology & Immunology, University ofSouthern California Keck School of Medicine, Los Angeles, California and Department of Pathology andLaboratory Medicine, University of PennsylvaniaSchool of Medicine, Philadelphia, Pennsylvania, USA

104. Clinical Trials

105. SB-728

112. Nat Biotech 2008

113. Nat Biotech 2008 25% CCR5 modification

116. About 50% of the human population has antibodies !

120. 6%, 3%, 1%, 2% and 2% at 14 days

127. Fever, Fatigue

129. Treatment interruption

132. 4%

147. Thank You