Engineering HIV-1-resistant T-cells from short-hairpin RNA-expressing hematopoietic stem/progenitor cells in humanized BLT mice.
Down-regulation of the HIV-1 coreceptor CCR5 holds significant potential for long-term protection against HIV-1 in patients. Using the humanized bone marrow/liver/thymus (hu-BLT) mouse model which allows investigation of human hematopoietic stem/progenitor cell (HSPC) transplant and immune system re...
Guardado en:
| Autores principales: | , , , , , , , , , , , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
Public Library of Science (PLoS)
2012
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/2b0ee1ff496b470290d5a8ee5f6b119e |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:2b0ee1ff496b470290d5a8ee5f6b119e |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:2b0ee1ff496b470290d5a8ee5f6b119e2021-11-18T08:02:58ZEngineering HIV-1-resistant T-cells from short-hairpin RNA-expressing hematopoietic stem/progenitor cells in humanized BLT mice.1932-620310.1371/journal.pone.0053492https://doaj.org/article/2b0ee1ff496b470290d5a8ee5f6b119e2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23300932/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Down-regulation of the HIV-1 coreceptor CCR5 holds significant potential for long-term protection against HIV-1 in patients. Using the humanized bone marrow/liver/thymus (hu-BLT) mouse model which allows investigation of human hematopoietic stem/progenitor cell (HSPC) transplant and immune system reconstitution as well as HIV-1 infection, we previously demonstrated stable inhibition of CCR5 expression in systemic lymphoid tissues via transplantation of HSPCs genetically modified by lentiviral vector transduction to express short hairpin RNA (shRNA). However, CCR5 down-regulation will not be effective against existing CXCR4-tropic HIV-1 and emergence of resistant viral strains. As such, combination approaches targeting additional steps in the virus lifecycle are required. We screened a panel of previously published shRNAs targeting highly conserved regions and identified a potent shRNA targeting the R-region of the HIV-1 long terminal repeat (LTR). Here, we report that human CD4(+) T-cells derived from transplanted HSPC engineered to co-express shRNAs targeting CCR5 and HIV-1 LTR are resistant to CCR5- and CXCR4- tropic HIV-1-mediated depletion in vivo. Transduction with the combination vector suppressed CXCR4- and CCR5- tropic viral replication in cell lines and peripheral blood mononuclear cells in vitro. No obvious cytotoxicity or interferon response was observed. Transplantation of combination vector-transduced HSPC into hu-BLT mice resulted in efficient engraftment and subsequent stable gene marking and CCR5 down-regulation in human CD4(+) T-cells within peripheral blood and systemic lymphoid tissues, including gut-associated lymphoid tissue, a major site of robust viral replication, for over twelve weeks. CXCR4- and CCR5- tropic HIV-1 infection was effectively inhibited in hu-BLT mouse spleen-derived human CD4(+) T-cells ex vivo. Furthermore, levels of gene-marked CD4(+) T-cells in peripheral blood increased despite systemic infection with either CXCR4- or CCR5- tropic HIV-1 in vivo. These results demonstrate that transplantation of HSPCs engineered with our combination shRNA vector may be a potential therapy against HIV disease.Gene-Errol E RingpisSaki ShimizuHubert ArokiumJoanna Camba-ColónMaria V CarrollRuth CortadoYiming XiePatrick Y KimAnna SahakyanEmily L LoweMunetoshi NarukawaFadi N KandarianBryan P BurkeGeoff P SymondsDong Sung AnIrvin S Y ChenMasakazu KamataPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e53492 (2012) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Gene-Errol E Ringpis Saki Shimizu Hubert Arokium Joanna Camba-Colón Maria V Carroll Ruth Cortado Yiming Xie Patrick Y Kim Anna Sahakyan Emily L Lowe Munetoshi Narukawa Fadi N Kandarian Bryan P Burke Geoff P Symonds Dong Sung An Irvin S Y Chen Masakazu Kamata Engineering HIV-1-resistant T-cells from short-hairpin RNA-expressing hematopoietic stem/progenitor cells in humanized BLT mice. |
| description |
Down-regulation of the HIV-1 coreceptor CCR5 holds significant potential for long-term protection against HIV-1 in patients. Using the humanized bone marrow/liver/thymus (hu-BLT) mouse model which allows investigation of human hematopoietic stem/progenitor cell (HSPC) transplant and immune system reconstitution as well as HIV-1 infection, we previously demonstrated stable inhibition of CCR5 expression in systemic lymphoid tissues via transplantation of HSPCs genetically modified by lentiviral vector transduction to express short hairpin RNA (shRNA). However, CCR5 down-regulation will not be effective against existing CXCR4-tropic HIV-1 and emergence of resistant viral strains. As such, combination approaches targeting additional steps in the virus lifecycle are required. We screened a panel of previously published shRNAs targeting highly conserved regions and identified a potent shRNA targeting the R-region of the HIV-1 long terminal repeat (LTR). Here, we report that human CD4(+) T-cells derived from transplanted HSPC engineered to co-express shRNAs targeting CCR5 and HIV-1 LTR are resistant to CCR5- and CXCR4- tropic HIV-1-mediated depletion in vivo. Transduction with the combination vector suppressed CXCR4- and CCR5- tropic viral replication in cell lines and peripheral blood mononuclear cells in vitro. No obvious cytotoxicity or interferon response was observed. Transplantation of combination vector-transduced HSPC into hu-BLT mice resulted in efficient engraftment and subsequent stable gene marking and CCR5 down-regulation in human CD4(+) T-cells within peripheral blood and systemic lymphoid tissues, including gut-associated lymphoid tissue, a major site of robust viral replication, for over twelve weeks. CXCR4- and CCR5- tropic HIV-1 infection was effectively inhibited in hu-BLT mouse spleen-derived human CD4(+) T-cells ex vivo. Furthermore, levels of gene-marked CD4(+) T-cells in peripheral blood increased despite systemic infection with either CXCR4- or CCR5- tropic HIV-1 in vivo. These results demonstrate that transplantation of HSPCs engineered with our combination shRNA vector may be a potential therapy against HIV disease. |
| format |
article |
| author |
Gene-Errol E Ringpis Saki Shimizu Hubert Arokium Joanna Camba-Colón Maria V Carroll Ruth Cortado Yiming Xie Patrick Y Kim Anna Sahakyan Emily L Lowe Munetoshi Narukawa Fadi N Kandarian Bryan P Burke Geoff P Symonds Dong Sung An Irvin S Y Chen Masakazu Kamata |
| author_facet |
Gene-Errol E Ringpis Saki Shimizu Hubert Arokium Joanna Camba-Colón Maria V Carroll Ruth Cortado Yiming Xie Patrick Y Kim Anna Sahakyan Emily L Lowe Munetoshi Narukawa Fadi N Kandarian Bryan P Burke Geoff P Symonds Dong Sung An Irvin S Y Chen Masakazu Kamata |
| author_sort |
Gene-Errol E Ringpis |
| title |
Engineering HIV-1-resistant T-cells from short-hairpin RNA-expressing hematopoietic stem/progenitor cells in humanized BLT mice. |
| title_short |
Engineering HIV-1-resistant T-cells from short-hairpin RNA-expressing hematopoietic stem/progenitor cells in humanized BLT mice. |
| title_full |
Engineering HIV-1-resistant T-cells from short-hairpin RNA-expressing hematopoietic stem/progenitor cells in humanized BLT mice. |
| title_fullStr |
Engineering HIV-1-resistant T-cells from short-hairpin RNA-expressing hematopoietic stem/progenitor cells in humanized BLT mice. |
| title_full_unstemmed |
Engineering HIV-1-resistant T-cells from short-hairpin RNA-expressing hematopoietic stem/progenitor cells in humanized BLT mice. |
| title_sort |
engineering hiv-1-resistant t-cells from short-hairpin rna-expressing hematopoietic stem/progenitor cells in humanized blt mice. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2012 |
| url |
https://doaj.org/article/2b0ee1ff496b470290d5a8ee5f6b119e |
| work_keys_str_mv |
AT geneerroleringpis engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT sakishimizu engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT hubertarokium engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT joannacambacolon engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT mariavcarroll engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT ruthcortado engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT yimingxie engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT patrickykim engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT annasahakyan engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT emilyllowe engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT munetoshinarukawa engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT fadinkandarian engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT bryanpburke engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT geoffpsymonds engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT dongsungan engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT irvinsychen engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice AT masakazukamata engineeringhiv1resistanttcellsfromshorthairpinrnaexpressinghematopoieticstemprogenitorcellsinhumanizedbltmice |
| _version_ |
1718422594480242688 |