The Canadian HIV Cure Enterprise (CanCURE) Team ric A. Cohen, PhD - - PowerPoint PPT Presentation

Éric A. Cohen, PhD

Laboratory of Human Retrovirology Institut de Recherches Cliniques de Montréal (IRCM)

How close are we to a cure?

HIV Endgame Conference, Toronto

October 25, 2016

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The Canadian HIV Cure Enterprise (CanCURE) Team

Disclosure

• Éric A. Cohen

I have no relationships with commercial interests to disclose

Outline

• 1. The Canadian HIV Cure Enterprise (CANCURE)
• 2. Context and rationale
• 3. CanCURE objectives and recent progress

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• CanCURE Team project was initiated

following a RFA on HIV Cure launched by the CIHR HIV/AIDS initiative in January 2013

• CanCURE is funded for 5-years (Jan 1, 2014-

Dec 31st, 2018) by the CIHR in partnership with CANFAR and IAS

• IRCM and the Université de Montréal are the

host institution http://www.cancurehiv.org

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CanCURE Mission

To understand HIV persistence during antiretroviral therapy (ART) and to harness this knowledge towards the development of HIV cure interventions

CanCURE Team

• 9 Principal Investigators

– P. Ancuta, CR-CHUM – J. Angel, U. of Ottawa – E.A. Cohen, IRCM – J. Estaquier, U. Laval – K. Fowkes, U. Manitoba – A. Mouland, McGill – M. Ostrowski, U. Toronto – J-P Routy, McGill – M.J. Tremblay, U. Laval

• 19 Co-Investigators

– B. Bell, U. Sherbrooke – J. Bell, U. Ottawa – R. Bendayan, U. Toronto – Z. Brumme, SFU – M. Brockman, SFU – C. Cheong, IRCM – A. Cochrane, U. Toronto – N. Chomont, CR-CHUM – A. Gatignol, McGill – É. Haddad, U. Montréal – D. Kaufmann, CR-CHUM – R. Kaul, U. Toronto – A. Kumar, U. Ottawa – M-A Langlois, U. Ottawa – T. Murooka, U. Manitoba – A. Poon, UBC – C. Power, U. Alberta – M. Wainberg, McGill – JC Zúñiga Pflücker, U. Toronto

• Community Liaison

– R. Reinhard

CanCURE Participating institutions consist of 10 Canadian Universities and affiliated research centers, including the IRCM, the Team Host Institution

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Context and Rationale

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Current HIV drugs do not eradicate HIV

8 Limit of detection

Circulating virus Time

START STOP

HAART

HIV hides in reservoirs that are not sensitive to current therapies

HIV infection is characterized by high levels of circulating viruses in the blood Antiretroviral drugs (HAART) are capable of suppressing HIV, even to undetectable levels However, the virus rebounds after cessation of therapy

Viral reservoirs represent the principal source of viral persistence during ART and a major obstacle to a cure

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Where does HIV persists?

10 Courtesy of Nicolas Chomont

HIV latency, a challenge for host immune defenses

11 Deeks et al., Nature Reviews Immunology 2012

HIV latency, a challenge for host immune defenses

12 Deeks et al., Nature Reviews Immunology 2012

Failure of effector cell clearance due to:

• Absence of viral protein expression
• Viral epitope escape
• Host immune exhaustion

Durable reservoir indifferent to treatment

• r to any host defense

targeting virus elements

While extensive efforts have been deployed in the direction of eliminating HIV-1 memory CD4+ T-cell, the predominant VR, much less is known about the contribution

• f myeloid cells and

particularly macrophages to the overall HIV reservoir It will be difficult to achieve a cure for HIV-1 without considering all potential VRs

Macrophages

• Found in virtually every tissue

in the body

• Originate from self-renewing

tissue-resident macrophages and infiltrating monocyte-derived macrophages.

• Maintain tissue homeostasis by

recognizing and disposing of apoptotic cells in a non pro- inflammatory manner

• Provide a critical front line of

defense against pathogens, including viruses by eliminating infected cells by phagocytosis

HIV-infected Macrophages

Verrolet et al. Blood , 2014

Macrophages as VR candidates

• Permissive to productive HIV

infection in vivo and in vitro (express CD4 and CCR5)

(Weinberg et al.,1991; Honeycutt et al., 2016)

• Harbor virus for long periods
• f time in intracellular virus-

containing compartments (VCC) (Groot et al., 2008)

• They are resistant to HIV-1-

induced apoptosis (Carter et al.,

2008)

• They can harbor virus in a

latent state or in a state of very low expression in vitro

(Kumar et al. Viruses, 2014)

Sattentau & Stevenson , 2016 Honeycutt et al., 2016

Hu-MoM

Macrophages as VR candidates

• Presence of proviral DNA

was detected in macrophages isolated from rectal and ileal tissue (Yukl et

al., 2014) as well as myeloid

cells isolated from GALT of ART-treated aviremic individuals (Josefsson et al, 2013)

• However, the fact that

macrophage can ingest infected CD4+ T cells (Baxter

et al., 2014) complicates the

interpretation

(Yukl et al. 2014) Sattentau & Stevenson 2016

Cure strategies

17 Courtesy Nicolas Chomont

CanCURE objectives and recent progress

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CanCURE Scientific Objectives

1) Identify, characterize and exploit insufficiently characterized properties of myeloid cells, especially macrophages, and additional lymphoid cell subsets within mucosal compartments that act as VRs; 2) Conduct detailed mechanistic studies aimed at understanding how these VRs are established and maintained; 3) Identify new drug candidates that reverse virus latency/persistence in multiple cross-acting VRs, and evaluate novel therapeutic strategies that enhance immune control and/or induce effective clearance of VRs; 4) Test whether immune-based therapies control or reduce VR in ART-treated HIV-infected patients in clinical trials

CanCURE Recent Progress

• HIV persists in CCR6+ CD4+ T cells from Colon and

Blood during antiviral Therapy (Gosselin/Wiche Salinas et al., AIDS, 2016, In Press)

• Single-cell characterization of viral translation-

competent reservoirs in HIV-Infected individuals (Baxter et al, Cell Host & Microbe, 2016)

• Enhancing virion tethering by BST2/Tetherin

sensitizes productively and latently HIV-infected T cells to ADCC mediated by broadly neutralizing antibodies (Pham et al., Scientific Reports)

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HIV-DNA Mainly Persists in Colon and Blood CCR6+ T-Cells during ART

21 Gosselin/Wiche-Salinas et al., AIDS, 2016, In Press

Blood Central Memory CCR6+ T-cells Are Enriched in Integrated HIV-DNA During ART

22 Gosselin/Wiche-Salinas et al., AIDS, 2016, In Press

Superior HIV Reactivation in CCR6+ Subsets during ART

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The preferential persistence of HIV in colon and blood CCR6+ T-cells during ART needs to be considered for tailored HIV eradication strategies

HIVRNA/Gag dual detection

Complete method

Day -1

CD4 T cell isolation and stimulation

Day 0

Surface and ICS antibody staining mRNA labelling

Day +1

Ampli cation and labelling Analysis CD4 isolation PBMCs Surface staining ICS for HIV-1 Gag protein L a b e l G a g P

• l

m R N A A m p l i f i c a t i

• n

2 Amplification 1 Label amplified probe Run on flow cytometer Rest or stimulate O/N

HIV mRNA

Store 4oC O/N + RNAsin

GagPol mRNA Gag Protein

2.8 15.1 0.4 81.7

Detection of translation-competent reservoirs

Baxter et al. Cell Host and Microbe 2016

This assay is currently adapted to examine HIV persistence in myeloid cells

IFNα Enhances Env Recognition and ADCC by PGT126

Pham et al.,

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…..In a BST2-Dependent Manner

Pham et al.,

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BST2 restriction is normally counteracted by the HIV-1 accessory protein

Enhancement of Virion Tethering by BST2 Sensitizes Reactivated Latent Cells to ADCC by pGT121

Pham et al.,

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bNAbs: PGT121

Restoring BST2 restriction could improve anti-HIV responses and potentially provide a means to eliminate reactivated cells in latent reservoirs

Thanks

IRCM

• Tram NQ Pham
• Sabelo Lukhele
• Mariana Bego
• Frédéric Dallaire
• Scott Sugden
• Mathieu Dubé

Reagents

• J. Robinson, Tulane (17b)
• M. Nussenzweig, Rockefeller (3BNC117)
• M. Connors, NIH (35O22; 7H6)
• D. Burton; P. Poignard , Scripps (PG9; PGT121;

PGT126)

• Idera Pharmaceuticals
• NIH AIDS Reagent Program
• IRCM Flow Cytometry Core

Healthy volunteers

Clinical Collaborators:

• P. Larochelle, M. Gauthier

and the IRCM Clinic staff Collaborators

• Jean-Pierre Routy, McGill,
• Élie Haddad, Université de Montréal,
• Winfried Weissenhorn, U. Grenoble-Alpes,
• Frank Kirchhoff, University of Ulm,
• Wei Cao, MD Anderson Cancer Center,
• Yong-Jun Liu, Sanofi
• Romas Geleziunas, Gilead
• Mario Legault, FRQ-S AIDS Network

CR-CHUM

• Amie Baxter
• Daniel Kaufmann
• Annie Gosselin
• Petronela Ancuta
• Nicolas Chomont

CanCURE

• Robert Reinhardt
• Sébastien Sabbagh

THANK YOU

CanCURE Mission

This collaborative effort engages basic and clinical scientists as well as members of the community in a research effort to:

• 1. Characterize mechanisms of HIV persistence in

the presence of ART

• 2. Develop cell-based and animal model systems in

which persistent infection can be investigated and therapeutic interventions can be tested.

• 3. Develop new assays to accurately characterize

and measure viral reservoirs

• 4. Generate therapeutic approaches that can

ultimately be tested in human clinical trials.

CanCURE Research Themes

• Theme 1: to study the molecular, genetics and functional

characteristics of HIV/SIV persistence in human and animal models (NHP and BLT mice) (Leaders: P. Ancuta & J. Estaquier)

• Theme 2: To define mechanisms governing HIV latency and

persistence in macrophages (Leaders: M.J Tremblay & A. Mouland)

• Theme 3: To identify new drug candidates and therapeutic

strategies aimed at eliminating HIV persistent infection and to test effective strategies in preclinical studies (Leaders: J. Angel & M. Ostrowski)

• Theme 4: To establish approaches, expertise and

infrastructure to conduct HIV Cure clinical trials by examining whether immune-based therapies reduce or eliminate VRs in ART-treated patients (Leaders: J-P Routy & J. Angel)

Scientific Advisory Board

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ERIC J. ARTS University of Western Ontario London, CANADA MICHAEL M. LEDERMAN Case Western Reserve University University Hospitals/Case Medical Center Cleveland, USA OLIVIER SCHWARTZ Institut Pasteur Paris, FRANCE GUIDO SILVESTRI Emory University Yerkes National Primate Research Center, Atlanta, USA

Community Advisory Board

• Robert Reinhard – CanCURE Community Liaison
• Shari Margolese – At Large and CTN
• Renée Masching – Canadian Aboriginal AIDS Network
• Tola Mbulaheni – African and Caribbean Council on

HIV/AIDS in Ontario

• Jonathan Postnikoff – Positive Living BC
• Ron Rosenes – At Large
• José Sousa – At Large
• Darien Taylor – At large
• Wangari Tharao – Women’s Health in Women’s Hands

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