eprintid: 1362851
rev_number: 44
eprint_status: archive
userid: 608
dir: disk0/01/36/28/51
datestamp: 2013-02-14 11:34:16
lastmod: 2019-10-19 08:44:57
status_changed: 2013-02-14 11:34:16
type: thesis
metadata_visibility: show
item_issues_count: 0
creators_name: Chan, E
title: Lentiviral gene therapy for HIV using TRIM-cyclophilin restriction factors
ispublished: unpub
divisions: A01
divisions: B02
divisions: D13
divisions: G24
abstract: Lentiviral vector delivery of anti-HIV elements could provide the basis of alternative therapies against HIV, potentially providing long term protection after a single intervention. Some primate species have evolved restriction factors formed by the fusion of TRIM5α and Cyclophilin A (TRIM5Cyp) following retrotransposition of CypA cDNA into the TRIM5 gene, which provide potent resistance against certain lentiviruses. We have designed humanised versions of these proteins combining both TRIM5 and TRIM21 with CypA, and investigated their potential for use in gene therapy against HIV-1. Both TRIM5- and TRIM21-Cyp fusion proteins provided strong restriction of HIV-1 in all of the systems tested, including primary human T cells. However, TRIM5Cyp was shown to disrupt the antiretroviral effect of endogenous TRIM5α and rescue murine retrovirus infection, whereas TRIM21Cyp caused no interference. In contrast, neither TRIM5CypA nor TRIM21CypA expression affected the antiviral activity of endogenous TRIM21.
In addition to TRIMCyp restriction factors, a second anti-HIV strategy was investigated using zinc finger nucleases (ZFNs) to knockout the HIV-1 co-receptor, CCR5. ZFNs introduce a double stranded break into the CCR5 gene, which can be restored by homology directed repair. Provision of a green fluorescent protein (GFP) or TRIM21Cyp donor template exploits this repair mechanism to allow site specific integration at the CCR5 locus, although at low efficiency. Using integrating vectors, we have shown that TRIMCyp mediated restriction is so potent that no additional inhibition was conferred by CCR5 knockout.
In conclusion, delivery of TRIMCyp genes using lentiviral vectors could form the basis of an intracellular vaccination strategy against HIV-1, with TRIM21Cyp having benefits by maintaining endogenous TRIM function. With further optimisation to improve efficiency, this could be combined with ZFNs for site specific integration of the transgene and knockout of CCR5 to provide a dual method of HIV-1 inhibition.
date: 2012-10-28
oa_status: green
thesis_class: doctoral_open
language: eng
thesis_view: UCL_Thesis
dart: DART-Europe
primo: open
primo_central: open_green
verified: verified_manual
elements_source: Manually entered
elements_id: 442686
lyricists_name: Chan, Emma
lyricists_id: ECHAN95
full_text_status: public
pagerange: ? - ?
pages: 232
institution: UCL (University College London)
department: ICH - Molecular Immunology Unit
thesis_type: Doctoral
citation:        Chan, E;      (2012)    Lentiviral gene therapy for HIV using TRIM-cyclophilin restriction factors.                   Doctoral thesis , UCL (University College London).     Green open access   
 
document_url: https://discovery.ucl.ac.uk/id/eprint/1362851/1/FinalEmmaChan.pdf