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Antibody engineering for HIV therapeutics : from fusion inhibition to delivery of genetic regulators
Publication . Santos, Ana Catarina Cunha, 1986-; Gonçalves, João, 1967-
Despite latest advances on antiretroviral therapeutics, HIV infection continues to be a chronic condition with serious complications and burden costs of treatment. The rapidly emergence of resistant strains constitutes a major contributor for failure of standard therapy. Accordingly, there is a continuous need for design of innovative HIV inhibitors against conserved and important targets on virus replication cycle. On the other hand, the exploration of novel classes of HIV inhibitors results in a necessity for novel efficient and specific delivery systems. In the present thesis, we manage to target HIV infection through two distinct therapeutic approaches based on single-domain antibody (sdAb) technology. In the first one, we developed a novel fusion inhibitor of HIV by rationally engineering the binding sites of a stable light-chain scaffold from rabbit-origin. From five sdAbs with specificity to the conserved and crucial-to-fusion N36 viral sequence, one potently inhibited HIV-1 infectivity. The selected fusion inhibitor, named F63, was capable of broadly inhibiting distinct strains of HIV-1 with an antiviral potency similar to the T-20 entry inhibitor, clinically approved for HIV/AIDS treatment. Moreover, F63 presented an effective neutralization activity against HIV type 2, which constitutes an advantage relative to the limited antiviral breadth of T-20. In the second approach, we took advantage of a previously validated variable domain from camel (nanobody) to design a strategy for CXCR4-targeted delivery of HIV expression inhibitors. The engineered nanobody-based vehicle delivered an anti-HIV small interfering RNA specifically to CXCR4-bearing cells—susceptible to HIV infection—being the activity of the delivered RNAi effector proved by virus promoter silencing and inhibition of HIV replication. A second version of this construct allowed to deliver a zinc-finger transcription factor, which functionality depends on CXCR4-mediated endocytosis as mechanism of cellular crossing-over. In resume, this study demonstrates the potential of engineering single-domain antibodies for HIV therapeutics and gives insights into the design of novel antiviral drugs.
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Fundação para a Ciência e a Tecnologia
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SFRH
Número da atribuição
SFRH/BD/73838/2010