DOCSLIB.ORG

Gene-Targeted Strategies to Eliminate HIV Latent Cells Through Synthetic Activators and Suicide Lentivectors

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

UNIVERSIDADE DE LISBOA Faculdade de Farmácia Gene-targeted strategies to eliminate HIV latent cells through synthetic activators and suicide lentivectors Pedro Ricardo Lucas Perdigão Orientadores: Prof. Doutor João Manuel Braz Gonçalves Doutora Mariana Canelhas Palminha Santa-Marta Tese especialmente elaborada para obtenção do grau de Doutor em Farmácia, especialidade Biotecnologia Farmacêutica. 2017 UNIVERSIDADE DE LISBOA Faculdade de Farmácia Gene-targeted strategies to eliminate HIV latent cells through synthetic activators and suicide lentivectors Pedro Ricardo Lucas Perdigão Orientadores: Prof. Doutor João Manuel Braz Gonçalves Doutora Mariana Canelhas Palminha Santa-Marta Tese especialmente elaborada para obtenção do grau de Doutor em Farmácia, especialidade Biotecnologia Farmacêutica. Júri: Presidente: Doutora Matilde da Luz dos Santos Duque da Fonseca e Castro, Professora Catedrática e Diretora da Faculdade de Farmácia da Universidade de Lisboa. Vogais: - Doutor Manuel António Faria Viola Gonçalves, Associate Professor Leiden University Medical Center, The Netherlands; - Doutor Luís Fernando Morgado Pereira Almeida, Professor Auxiliar Faculdade de Farmácia da Universidade de Coimbra; - Doutor Nuno Eduardo Moura dos Santos da Costa Taveira, Professor Catedrático Instituto Superior de Ciências da Saúde Egas Moniz; - Doutora Ana Cristina Gomes Espada de Sousa, Investigadora Coordenadora Faculdade de Medicina da Universidade de Lisboa; - Doutor José António Frazão Moniz Pereira, Professor Catedrático Faculdade de Farmácia da Universidade de Lisboa; - Doutor João Manuel Braz Gonçalves, Professor Associado com Agregação Faculdade de Farmácia da Universidade de Lisboa, Orientador. Fundação para a Ciência e Tecnologia – Ministério da Educação e Ciência: SFRH/BD/81941/2011 2017 Pedro Ricardo Lucas Perdigão was financially supported by a PhD fellowship (SFRH/BD/81941/2011) from Fundação para a Ciência e a Tecnologia—Ministério da Educação e Ciência (FCT-MEC), Portugal. This work was supported by grants from Alto Comissariado para a Saúde and Fundação para a Ciência e Tecnologia - Ministério da Educação e Ciência (FCT-MEC) (VIH/SAU/0013/2011, VIH/SAU/0020/2011 and HIVERA/0002/2013), The Skaggs Institute for Chemical Biology and the National Institutes of Health (DP1CA174426) and Bill and Melinda Gates Foundation (Grand Challenges Explorations Round 7). The research described in the present thesis was performed from January 2012 until March 2017 under the supervision of Professor João Gonçalves and Dr. Mariana-Santa-Marta at the URIA- Unidade de Retrovírus e Infeções Associadas and Research Institute for Medicines (iMed.ULisboa) of the Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal. This project was also performed in collaboration with Professor Carlos F. Barbas III from the The Skaggs Institute for Chemical Biology and Departments of Chemistry and Cell and Molecular Biology of The Scripps Research Institute, La Jolla, California, United States of America. The results described in this thesis were included in manuscripts already published or in preparation, in peer-reviewed journals and oral or poster communications of international and national scientific meetings: Articles in peer review journals Perdigão P, Cunha-Santos C, Barbas III C, Santa-Marta M, Gonçalves J. Short-term protein delivery of cell-penetrating zinc-finger activators stimulate latent HIV-1 expression. Manuscript in preparation. Perdigão P, Gaj T, Santa-Marta M, Barbas III C, Gonçalves J (2016). Reactivation of latent HIV-1 expression by engineered TALE transcription factors. Plos One, 2;11(3):e0150037. Oral communications Perdigão P, Gaj T, Rocha C, Barbas III C, Santa-Marta M, Gonçalves J. Gene-therapeutic approach complementing TALE activators and suicidal vectors target and eliminate HIV latent cells. 8th iMed.UL Postgraduate Students Meeting 2016, Lisboa, Portugal, OC11, pp 12. Perdigão P, Gaj T, Rocha C, Barbas III C, Santa-Marta M, Gonçalves J. Gene-therapeutic approach complementing TALE activators and suicidal vectors target and eliminate HIV latent cells. FASEB Science Research Conference: Genome Engineering - Cutting-Edge Research and Applications 2016, Lisbon, Portugal. Perdigão P, Gaj T, Rocha C, Barbas III C, Santa-Marta M, Gonçalves J. Specific elimination of latent HIV infected cells through a “shock and kill” suicidal gene therapy. 8th Biennal Congress of the Spanish Society of Gene and Cell Therapy 2015, San Sebastian, Spain. Posters in conferences Perdigão P, Gaj T, Rocha C, Barbas III C, Santa-Marta M, Gonçalves J. Gene-therapeutic approach complementing TALE activators and suicidal vectors target and eliminate HIV latent cells. FASEB Science Research Conference: Genome Engineering - Cutting-Edge Research and Applications 2016, Lisbon, Portugal. Perdigão P, Gaj T, Barbas III C, Santa-Marta M, Gonçalves J. Reactivation of latent HIV-1 expression by engineered TALE transcription factors. ESGCT and FSGT Collaborative Congress 2015, Helsinki, Finland. Perdigão P, Gaj T, Barbas III C, Santa-Marta M, Gonçalves J. Reactivation of latent HIV-1 expression by engineered TALE transcription factors. 7th iMed.UL Postgraduate Students Meeting 2015, Lisboa, Portugal. Perdigão P, Gaj T, Barbas III C, Santa-Marta M, Gonçalves J. Engineered artificial transcription factors as novel tools to eliminate HIV latency. 6th iMed.UL Postgraduate Students Meeting 2014, Lisboa, Portugal. Perdigão P, Santa-Marta M, Gonçalves J. Specific elimination of HIV-1 infected cells using a Tat/Rev-dependent circuit, A Collaborative Solution for Tropical Diseases: The Luso- American Response. Conference on HIV/AIDS research 2011, Lisboa, Portugal. Agradecimentos AGRADECIMENTOS Em primeiro lugar, quero agradecer aos meus orientadores. Ao Professor Doutor João Gonçalves por me receber no seu laboratório, por me ter introduzido nesta área tão fascinante da terapia génica e engenharia do genoma, pela sua orientação ao longo deste trabalho, pela confiança que teve em mim e pelo incentivo. À Doutora Mariana Santa-Marta pela orientação, paciência e acompanhamento desde o início da minha carreira como investigador, por todos os conhecimentos e conselhos transmitidos, e por todo o apoio dado não só no trabalho, mas também a nível pessoal. Gostaria de agradecer também ao Professor Doutor Moniz-Pereira, por me ter acolhido no seu departamento. Quero também fazer um agradecimento especial ao falecido Professor Doutor Carlos Barbas, por me ter aceite no seu laboratório no Instituto Scripps. A sua morte foi também uma grande perda para a comunidade científica que ficou privada de um brilhante investigador que ainda tinha tanto para oferecer. Infelizmente não pude interagir diretamente consigo e aprender com a sua experiência, no entanto disponibilizou todos os meios e conhecimentos do seu laboratório e como tal teve um enorme contributo para a conclusão deste trabalho. Obrigado! Quero agradecer aos meus colegas do JG lab que se cruzaram comigo durante o meu percurso. Obrigado Catarina (companheira em todos os momentos), Soraia, Cátia e Ana Catarina pela vossa amizade, apoio e companhia ao longo de todos estes anos. Obrigado Paula, Luís, Vasco, Lídia, Andreia, Acilino, Inês, Sofia Romano, Cheila, Renato, Sofia Cerqueira, Joana, Sofia Narciso, Sylvie, Rita e André (espero não me esquecer de ninguém). Todos contribuíram de alguma forma para meu percurso. Obrigado por toda a disponibilidade dada e paciência que tiveram para me transmitir conhecimentos ou pelos momentos de companheirismo e descontração que ajudam a ultrapassar os dias mais difíceis. Obrigado Catarina e Cheila pela vossa participação direta neste trabalho. Obrigado também aos membros do laboratório do Professor Nuno Taveira e Professora Cecília Rodrigues, pela amizade e partilha de conhecimento e reagentes. Obrigado ao Professor João Barata por disponibilizar o seu laboratório para experiências. I would like to thank the members of the Barbas lab, for taking me in and for your support and friendship. You were essential for my integration into your group. I would like to thank Dr. Joel Gottesfeld for taking the role of Dr. Barbas and supervising my work during our labmeetings, for your mentorship as well for your sympathy. Thank you, Roberta and Mishelle i Agradecimentos for your efforts to keep the lab running and organized, and for providing me with all the conditions that I could wish for to carry out my projects. Thank you, Thom, for your friendship and for your help and advice that helped me grow as a researcher. Thank you, Jia, for sharing your knowledge as well and for your support. Thanks Shannon and Mike Z. for all the good times. I miss you all and hope to see you again! Thank you, Brian, Klaus, Mark, Wei, Robyn, James and all the remaining members of the lab for your assistance and friendship. Finally, I would also like to thank to the ones that took me into their houses and shared their lives during my stay at San Diego, John, Paul, Fred, Aaron and Patrick. Thank you all! Aos meus amigos, pelos momentos de diversão que sempre conseguiram colocar um sorriso na minha cara, mesmo nos momentos mais complicados. Obrigado por todo o incentivo! Quero agradecer à minha família, aos meus pais por todo o amor e compreensão. Obrigado por acreditarem sempre em mim e no meu percurso. Sei que têm orgulho em mim e sempre me esforcei para corresponder o melhor possível a toda a educação e apoio que me deram. Ao meu irmão, por ser sempre um exemplo e me incentivar a ser melhor. À minha cunhada, por todo apoio dado e pela
Recommended publications
  • Mechanisms of HIV Persistence in HIV Reservoirs

    Mechanisms of HIV Persistence in HIV Reservoirs

    Mechanisms of HIV persistence in HIV reservoirs 1. Mayibongwe Louis Mzingwane* Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, P. Bag X323, ARCADIA 0007, Pretoria, South Africa Department of Pathology, Faculty of Medicine, National University of Science and Technology, P. O Box AC 939, Ascot, Bulawayo, Zimbabwe Email: [email protected] 2. Caroline T. Tiemessen Centre for HIV and Sexually Transmitted Infections, National Institute of communicable Diseases, Private Bag X4, Sandringham, 2131, Johannesburg, South Africa Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa Email: [email protected] *Corresponding author Keywords: HIV reservoirs, persistent viremia Running head: HIV compartments and persistence 1 Summary The establishment and maintenance of HIV reservoirs which lead to persistent viremia in patients on antiretroviral (ARV) drugs remains the greatest challenge of the highly active antiretroviral therapy (HAART) era. Cellular reservoirs include resting memory CD4+ T lymphocytes, these implicated as the major HIV reservoir, having a half-life of approximately 44 months while this is less than 6 hours for HIV in plasma. In some individuals persistent viremia consists of invariant HIV clones not detected in circulating resting CD4+ T lymphocytes suggesting other possible sources of residual viremia. Some anatomical reservoirs that may harbour such cells include the brain and the central nervous system (CNS), the gastrointestinal tract (GIT) and the gut-associated lymphoid tissue (GALT) and other lymphoid organs and the genital tract. The presence of immune cells and other HIV susceptible cells, occurring in differing compositions in anatomical reservoirs, coupled with variable and poor drug penetration which results in suboptimal drug concentrations in some sites, are all likely factors that fuel the continued low level replication and persistent viremia during treatment.
  • Directed Evolution of an HIV-1 LTR Specific Recombinase for Anti-Retroviral Therapy- a Proof of Concept Study

    Directed Evolution of an HIV-1 LTR Specific Recombinase for Anti-Retroviral Therapy- a Proof of Concept Study

    Directed evolution of an HIV-1 LTR specific recombinase for anti-retroviral therapy- a proof of concept study. DISSERTATION zur Erlangung des akademischen Grades Doctor rerum naturalium (Dr. rer. nat.) vorgelegt der Fakultät Mathematik und Naturwissenschaften der Technischen Universität Dresden von INDRANI SARKAR geboren am 03.12.1974, Kolkata, INDIA. Gutachter: Dr. Frank Buchholz Prof. Dr. Francis Stewart Prof. Dr. Joachim Hauber Eingereicht am 14.06.2006 Tag der Verteidigung: i Acknowledgements First and foremost I would like to thank my supervisor, Frank Buchholz for giving me the opportunity to work in his group and for motivating and supporting me through all these years. I am extremely grateful to him for having unlimited patience especially during the long evolution phase of this project and for always giving me helpful advice. He has been very supportive and reassuring in every situation. I am also thankful to him for the great lab retreats he organized. I would like to thank my TAC committee member Francis Stewart for his helpful suggestions. I am grateful to Joachim Hauber and Ilona Hauber (Heinrich-Pette Institute, Hamburg) who collaborated with me on this project and helped me to develop this project and provided me with the HIV-1 LTR based vectors and cell lines. I would like to thank Bianca Habermann for modeling of mutations and also Mayte Pissabarro with whom I had first looked into the recombinase mutations. A special thanks to all my lab members who made the lab a nice place to work in and also MPI-CBG for the friendly and cooperative atmosphere.
  • HIV Cure Research: Advances and Prospects

    HIV Cure Research: Advances and Prospects

    HIV cure research: advances and prospects. Caroline P Passaes, Asier Sáez-Cirión To cite this version: Caroline P Passaes, Asier Sáez-Cirión. HIV cure research: advances and prospects.. Virology, Elsevier, 2014, 454-455, pp.340-52. 10.1016/j.virol.2014.02.021. pasteur-01420527 HAL Id: pasteur-01420527 https://hal-pasteur.archives-ouvertes.fr/pasteur-01420527 Submitted on 24 Apr 2017 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution - NonCommercial - ShareAlike| 4.0 International License HIV cure research: advances and prospects Caroline P. Passaesa,b, Asier Saez-Ciriona a Unité de Régulation des Infections Rétrovirales, Institut Pasteur, 25–28 rue du Dr Roux, 75724 Paris Cedex 15, France. b CEA, Division of Immuno-Virology, iMETI/DSV, 18 Route du Panorama, 92265 Fontenay-aux-Roses, France. Email addresses: [email protected]/[email protected] [email protected] Corresponding author: Asier Saez-Cirion Unité de Régulation des Infections Rétrovirales - Institut Pasteur 25–28 rue du Dr Roux, 75724 Paris Cedex 15, France Tel: +33 145688944; Fax: +33 145688957 E-mail: [email protected] 1 Abstract Thirty years after the identification of HIV, a cure for HIV infection is still to be achieved.
  • CONFERENCE CAROUSEL Editor’S Letter

    CONFERENCE CAROUSEL Editor’S Letter

    MARCH-APRIL 2012 IAVIReportThe Publication on AIDS Vaccine Research WWW.IAVIREPORT.ORG | VOLUME 16, NUMBER 2 CROI AND KEYSTONE: CONFERENCE CAROUSEL EDITor’s lETTER When managing editor Kristen Jill Kresge informed her staff that she was leaving the editing of IAVI Report in my hands while she went off to have a baby, I imagine their response was, well, polite. Or maybe not: Kristen is, as subscribers to this magazine doubtless know, a tough act to follow, even temporarily. Fortunately for me, she had the grace—and good sense—to plan out the issue before she went on leave. Aware that major HIV conferences were looming, Kristen dispatched her reporters to cover them. You will notice that they did so with gusto. Our sweeping report from the 19th Conference on Retrovi- ruses and Opportunistic Infections (CROI) may not qualify as light summer reading, but it does have a little something for everybody (see page 4). It recounts how researchers are harnessing new technolo- gies to solve the structure of the recalcitrant HIV spike, and shares insights gleaned from the continu- ing analysis of samples collected during the RV144 trial, which demonstrated for the first time that a wvaccine can prevent HIV transmission. Another segment delves into similar analyses of trials evaluat- ing antiretroviral treatment as a mode of prevention. For those with an appetite for basic biology, the article includes absorbing descriptions of HIV’s interaction with an elusive subclass of T helper cells, and research parsing the evolution of HIV’s intracellular defenses. Our articles on the Keystone Symposia this season are much more focused.
  • Highlights from the 2014 International Symposium on HIV

    Highlights from the 2014 International Symposium on HIV

    Lafeuillade et al. AIDS Research and Therapy 2014, 11:28 http://www.aidsrestherapy.com/content/11/1/28 RESEARCH Open Access Highlights from the 2014 International Symposium on HIV & Emerging Infectious Diseases (ISHEID): from cART management to the end of the HIV pandemic Alain Lafeuillade1*, Mark Wainberg2, Marie-Lise Gougeon3, Sabine Kinloch-de Loes4, Philippe Halfon5 and Hervé Tissot-Dupont6 Abstract The 2014 International Symposium on HIV and Emerging Infectious Diseases (ISHEID) provided a forum for investigators to hear the latest research developments in the clinical management of HIV and HCV infections as well as HIV cure research. Combined anti-retroviral therapy (c-ART) has had a profound impact on the disease prognosis and transformed this infection into a chronic disease. However, HIV is able to persist within the infected host and the pandemic is still growing. The main 2014 ISHEID theme was, hence “Together for a world without HIV and AIDS”. In this report we not only give details on this main topic but also summarize what has been discussed in the areas of HCV coinfection and present a short summary on currently emerging viral diseases. Keywords: HIV, HCV, Hepatitis E, Antiretroviral drugs, HIV vaccine, HIV cure, HIV reservoirs Introduction The conference started by “The Thirty Years of Dis- The 2014 International Symposium on HIV and Emerging coveries of HIV” and “The Ten Big Mistakes We Did”. Infectious Diseases (ISHEID) was held in Marseille, Stefano Vella reminded us of the hopes in the early days France, on May 21-23. This meeting attracted more than of the epidemic of a prophylactic vaccine within the next 900 delegates from all over the world (75% outside France) 3 years.
  • Abstracts from the International Symposium HIV & Emerging Infectious Diseases 2014 Marseille, France

    Abstracts from the International Symposium HIV & Emerging Infectious Diseases 2014 Marseille, France

    BMC Infectious Diseases 2014, Volume 14 Suppl 2 http://www.biomedcentral.com/bmcinfectdis/supplements/14/S2/ MEETING ABSTRACTS Open Access Abstracts from the International Symposium HIV & Emerging Infectious Diseases 2014 Marseille, France. 21-23 May 2014 Published: 23 May 2014 These abstracts are available online at http://www.biomedcentral.com/bmcinfectdis/supplements/14/S2/ Conclusion: UDS was more sensitive than the standard Sanger ORAL PRESENTATIONS sequencing. A combination of UDS and DeepChek® software for the interpretation of DR results saved a considerable amount of time and would help clinicians provide suitable treatments. A cut-off of 1% allowed MANAGING ANTIRETROVIRAL THERAPY, a better characterization of the viral population by identifying additional PART 1 resistance mutations and improving the DR interpretation. O1 Clinical impact of ultra deep versus Sanger sequencing detection of O2 minority mutations on HIV-1 drug resistance genotype interpretation First-line therapy with LPV/r vs NVP and 2 NRTIs in a developing after virological failure country: W144 of a randomized trial Sofiane Mohamed1,4*, Guillaume Penaranda1, Dimitri Gonzalez2, Claire Camus1, Nathan Clumeck*, Claude Mwamba, Kabamba Kabeya, Vincent Calvez, Hacène Khiri1, Ronan Boulmé2, Chalom Sayada2, Patrick Philibert3, Serge Matanda, Dolorès Vaira, Gilles Peytavin, Coca Necsoi, Marc Delforge, Daniel Olive4, Philippe Halfon1 David Kadiebwe, Chantal Milolo, Joe Ilunga, Liévin Kapend 1Alphabio Laboratory, Marseille, France; 2Advanced Biological Laboratories Saint Pierre University Hospital, Brussels, Belgium (ABL), Luxembourg, Luxembourg; 3European Hospital, Marseille, France; BMC Infectious Diseases 2014, 14(Suppl 2):O2 4Paoli Calmettes Institute, Tumor Immunology Laboratory and INSERM Center of Cancer Research, Marseille, France Aim: In resource-limited countries, NNRTI-based regimen may result in BMC Infectious Diseases 2014, 14(Suppl 2):O1 emergence of more HIV drug resistance because of a low genetic barrier.
  • (12) United States Patent (10) Patent No.: US 8,871,516 B2 Hauber Et Al

    (12) United States Patent (10) Patent No.: US 8,871,516 B2 Hauber Et Al

    USOO887.1516 B2 (12) United States Patent (10) Patent No.: US 8,871,516 B2 Hauber et al. (45) Date of Patent: Oct. 28, 2014 (54) USE OF TAILORED RECOMBINASES FOR (56) References Cited THE TREATMENT OF RETROVRAL INFECTIONS U.S. PATENT DOCUMENTS 6,262,341 B1 7/2001 Baszczynski et al. (75) Inventors: Joachim Hauber, Hamburg (DE); 6,890,726 B1 5/2005 Sauer et al. Frank Buchholz, Dresden (DE); Ilona 7,112,715 B2 9, 2006 Chambon et al. Hauber, Hamburg (DE); Francis A 2006, OO14264 A1 1/2006 Sauer et al. Stewart, Dresden (DE); Indrani 2009/0217400 A1* 8/2009 Carmi et al. .................... 800, 14 Sarkar, Kolkata (IN) FOREIGN PATENT DOCUMENTS (73) Assignees: Technische Universität Dresden, WO O2/44409 A2 6, 2002 Dresden (DE); WO 2005/081632 A2 * 9, 2005 Max-Planck-Gesellschaft Zur WO 2008/083931 A1 T 2008 Forderung der Wissenschaften E.V. OTHER PUBLICATIONS München (DE); Heinrich-Pette-Institut fir Experimentelle Virologie und Abremski et al., “Studies on the Properties of P1 Site-Specific Immunologie an der Universität Recombination: Evidence for Topologically Unlinked Products fol Hamburg, Hamburg (DE) lowing Recombination.” Cell 32: 1301-1311, Apr. 1983. Abremski et al., “Bacteriophage P1 Site-specific Recombination— (*) Notice: Subject to any disclaimer, the term of this Purification and Properties of the Cre Recombinase Protein.” J. Biol. patent is extended or adjusted under 35 Chem. 259(3): 1509-1514, Feb. 10, 1984. U.S.C. 154(b) by 651 days. Adachi et al., “Production of Acquired Immunodeficiency Syn drome-Associated Retrovirus in Human and Nonhuman Cells (21) Appl.
  • New Treatments and Targets

    New Treatments and Targets

    NEW TREATMENTS AND TARGETS P174 Universal Tre (uTre) recombinase specifically targets the majority of HIV-1 isolates Karpinski, J*; Chemnitz, J; Hauber, I; Abi-Ghanem, J; Paszkowski-Rogacz, M; Surendranath, V; Chakrabort, D; Hackmann, K; Schröck, E; Pisabarro, M; Hauber, J; Buchholz, F (Dresden, Germany) P175 Targeted destruction of HIV positive cells Sharma, J*; Dodgen, C; Skepu, A; Meyer, M (Cape Town, South Africa) *Indicates presenting author. Universal Tre (uTre) recombinase specifically targets the majority of HIV-1 isolates Janet Karpinski1, Jan Chemnitz2, Ilona Hauber2, Helga Hofmann-Sieber2, Carola Schäfer2, Niklas Beschorner2, Josephine Abi-Ghanem3, Maciej Paszkowski- Rogacz1, Vineeth Surendranath4, Debojyoti Chakraborty1, Karl Hackmann5, Evelin Schröck5, María Teresa Pisabarro3, Joachim Hauber2, Frank Buchholz1,4 1Department of Medical Systems Biology, Medical Faculty Carl Gustav Carus, TU Dresden, Germany, 2Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany, 3Department of Structural Bioinformatics, TU Dresden, Germany, 4Max-Planck-Institute for Molecular Cell Biology and Genetics, Dresden, Germany, 5Institute for Clinical Genetics, Medical Faculty Carl Gustav Carus, TU Dresden, Germany ABSTRACT Background: Current drugs against HIV can suppress the progression to AIDS but Methods: We employed the search tool SeLOX [3] in Results: Highly specific enzymatic activity on loxLTRu is demonstrated for cannot clear the patient from the virus. Because of potential side effects of these drugs order to find a well-conserved HIV-1 proviral sequence uTre in both Escherichia coli and human cells. Naturally occurring viral variants and the possible development of drug resistance, finding a cure for HIV infection that could serve as target site for a universal Tre from with single mutations within the loxLTRu sequence are also shown to be remains a high priority of HIV/AIDS research.
  • NIH Public Access Author Manuscript Clin Pharmacol Ther

    NIH Public Access Author Manuscript Clin Pharmacol Ther

    NIH Public Access Author Manuscript Clin Pharmacol Ther. Author manuscript; available in PMC 2014 March 05. NIH-PA Author ManuscriptPublished NIH-PA Author Manuscript in final edited NIH-PA Author Manuscript form as: Clin Pharmacol Ther. 2013 January ; 93(1): 46–56. doi:10.1038/clpt.2012.202. Prospects for Treatment of Latent HIV KM Barton1, BD Burch2, N Soriano-Sarabia2, and DM Margolis1,2,3 1Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA 2Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA 3Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA Abstract Recent advances in antiretroviral therapy (ART) have drastically improved the quality of life for people with HIV infection. However, owing to the persistence of latent HIV in the presence of therapy, patients must remain on therapy indefinitely. Currently, the solution to the HIV pandemic rests on the prevention of new infections and many decades of ART for the steadily expanding number of people infected worldwide. ART is costly, requires ongoing medical care, and can have side effects, thereby preventing its universal availability. Therefore, to escape the ironic burdens of therapy, efforts have begun to develop treatments for latent HIV infection. Current approaches propose either complete eradication of infection or induction of a state of stringent control over viral replication without ART. This review will discuss these strategies in detail and their potential for clinical development. More than 30 million people are currently infected with HIV, and more than two million became newly infected in 2010.1 Significant progress has been made in the treatment of HIV infection, and current therapies allow infected individuals to attain near-normal life spans.
  • Tat-Independent Lentivirus Genomes for Vaccination and Host/Pathogen Interaction Studies Deepanwita Bose

    Tat-Independent Lentivirus Genomes for Vaccination and Host/Pathogen Interaction Studies Deepanwita Bose

    Tat-independent lentivirus genomes for vaccination and host/pathogen interaction studies Deepanwita Bose To cite this version: Deepanwita Bose. Tat-independent lentivirus genomes for vaccination and host/pathogen interac- tion studies. Human health and pathology. Université Grenoble Alpes, 2017. English. NNT : 2017GREAV009. tel-01938074 HAL Id: tel-01938074 https://tel.archives-ouvertes.fr/tel-01938074 Submitted on 28 Nov 2018 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THÈSE Pour obtenir le grade de DOCTEUR DE LA COMMUNAUTÉ UNIVERSITÉ GRENOBLE ALPES Spécialité : Virologie/Microbiologie/Immunologie Arrêté ministériel : 7 août 2006 Présentée par Deepanwita BOSE Thèse dirigée par Dr. Yahia Chebloune préparée au sein du Laboratoire Pathogénèse et Vaccination Lentivirales (PAVAL Lab) dans l'École Doctorale Chimie & Sciences du Vivant (ED218) Tat-independent lentivirus genomes for vaccination and host/pathogen interaction studies. Thèse soutenue publiquement le 26 Janvier 2017, devant le jury composé de : Dr. Giuseppe Bertoni Maitre de conférences, IVI, Bern, Suisse Rapporteur Pr. Francois Villinger Professeur, Director of New Iberia Research Center, USA, Rapporteur Pr. Philippe Sabatier Professeur à l’Université Grenoble Alpes, Président Dr.Yahia Chebloune Research Director, à l’Université Grenoble Alpes, Examinateur Dr.
  • A M S J HIV/AIDS: Let’S See How Far We’Ve Come

    A M S J HIV/AIDS: Let’S See How Far We’Ve Come

    Review Article A M S J HIV/AIDS: let’s see how far we’ve come Lauren Fowler Lauren is a third year medical student at Griffith University. She has an undergraduate degree Third Year Medicine (Undergraduate) in science from the University of Sydney, and completed immunology honours in HIV retroviral Griffith University genetics, studying microRNA changes during HIV infection. She has a passion for the intricacies BSc (Hons) of the immune system and the pursuit of a cure for HIV. Now more than ever, HIV positive people are living longer and healthier lives because of access to antiretroviral therapy. Healthcare organisations are working to ensure that HIV positive people all over the world have access to the medical care they need to stay healthy. In the last few years, research into vaccine development, genetics-based approaches and novel therapies have achieved some progress and drug therapy regimens have become more effective. Public health strategies have aimed to reduce transmission, and early access to treatment has dramatically improved quality of life. With resources and funding aimed in the right directions, it will be possible to continue making significant progress towards better prevention, improved treatment options and perhaps even a cure for HIV and the elimination of the global AIDS epidemic. This article reviews some of the successes and difficulties in the scientific, research and treatment arm of infection progression to AIDS, and divides the infection into stages combating the HIV epidemic. There is still much work to be done, reflecting the decline in immune function. [11] The stages of infection but for now, let’s see how far we’ve come.
  • Directed Evolution of a Recombinase That Excises the Provirus of Most

    Directed Evolution of a Recombinase That Excises the Provirus of Most

    ARTICLES Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity Janet Karpinski1,2,11, Ilona Hauber2,11, Jan Chemnitz2,11, Carola Schäfer2,3, Maciej Paszkowski-Rogacz1, Deboyoti Chakraborty1, Niklas Beschorner2, Helga Hofmann-Sieber2,3, Ulrike C Lange2–4, Adam Grundhoff2,3, Karl Hackmann5, Evelin Schrock5, Josephine Abi-Ghanem6, M Teresa Pisabarro6, Vineeth Surendranath7, Axel Schambach8, Christoph Lindner9, Jan van Lunzen2,3,10, Joachim Hauber2,3 & Frank Buchholz1,7 Current combination antiretroviral therapies (cART) efficiently suppress HIV-1 reproduction in humans, but the virus persists as integrated proviral reservoirs in small numbers of cells. To generate an antiviral agent capable of eradicating the provirus from infected cells, we employed 145 cycles of substrate-linked directed evolution to evolve a recombinase (Brec1) that site- specifically recognizes a 34-bp sequence present in the long terminal repeats (LTRs) of the majority of the clinically relevant HIV-1 strains and subtypes. Brec1 efficiently, precisely and safely removes the integrated provirus from infected cells and is efficacious on clinical HIV-1 isolates in vitro and in vivo, including in mice humanized with patient-derived cells. Our data suggest that Brec1 has potential for clinical application as a curative HIV-1 therapy. Although substantial advances have been made in the treatment of HIV We now report the development and application of the broad-range infection using cART, present therapies do not cure HIV-infected indi- recombinase 1 (Brec1) that is active against the majority of HIV-1 viduals1–3. A small population of latent, transcriptionally silent but still primary isolates.