Limits...
Nanotherapeutics Using an HIV-1 Poly A and Transactivator of the HIV-1 LTR-(TAR-) Specific siRNA.

Mahajan SD, Aalinkeel R, Reynolds JL, Nair B, Sykes DE, Law WC, Ding H, Bergey EJ, Prasad PN, Schwartz SA - Patholog Res Int (2011)

Bottom Line: We used a well-validated siRNA (si510) which targets the poly A/TAR (transactivator of the HIV-1 LTR) site and suppresses viral replication.Our results suggest that the QR-si510 HIV-1 siRNA nanoplex is not only effective in delivering siRNA, but also in suppressing HIV-1 viral replication for a longer time period.HIV-1 nanotherapeutics can thus enhance systemic bioavailability and offer multifunctionality.

View Article: PubMed Central - PubMed

Affiliation: Division of Allergy, Immunology, and Rheumatology, Department of Medicine, University at Buffalo, The State University of New York, 640 Ellicott Street, Room 444 Innovation Center, Buffalo, NY 14203, USA.

ABSTRACT
HIV-1 replication can be efficiently inhibited by intracellular expression of an siRNA targeting the viral RNA. We used a well-validated siRNA (si510) which targets the poly A/TAR (transactivator of the HIV-1 LTR) site and suppresses viral replication. Nanotechnology holds much potential for impact in the field of HIV-1 therapeutics, and nanoparticles such as quantum rods (QRs) can be easily functionalized to incorporate siRNA forming stable nanoplexes that can be used for gene silencing. We evaluated the efficacy of the QR-si510 HIV-1 siRNA nanoplex in suppressing viral replication in the HIV-1-infected monocytic cell line THP-1 by measuring p24 antigen levels and gene expression levels of HIV-1 LTR. Our results suggest that the QR-si510 HIV-1 siRNA nanoplex is not only effective in delivering siRNA, but also in suppressing HIV-1 viral replication for a longer time period. HIV-1 nanotherapeutics can thus enhance systemic bioavailability and offer multifunctionality.

No MeSH data available.


Related in: MedlinePlus

Effect of QR-si510 siRNA nanoplex on LTR/RU5 gene expression in HIV-1-infected THP-1 cells. THP-1 monocytic cultures were treated with lipofectamine transfected scrambled siRNA (10 nM), Lipofectamine transfected si510 HIV-1 siRNA (10 nM) and  the QR-si510 siRNA (10 nM) nanoplex for time period ranging from 12 hr to 1 week posttransfection. At the end of the incubation period, RNA was extracted, reverse transcribed and the LTR/RU5 gene expression quantitated using Q-PCR. Our results show a significant decrease in HIV-1 LTR gene expression in HIV-1-infected THP-1 cells treated with the 10 nM QR-si510 HIV-1siRNA nanoplex at 12, 24, 48, 96 hr, and 1 week posttransfection as compared to the untransfected THP-1 cells and  the scrambled control.  The results shown are mean ± SD of 3 separate experiments done in duplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3108557&req=5

fig5: Effect of QR-si510 siRNA nanoplex on LTR/RU5 gene expression in HIV-1-infected THP-1 cells. THP-1 monocytic cultures were treated with lipofectamine transfected scrambled siRNA (10 nM), Lipofectamine transfected si510 HIV-1 siRNA (10 nM) and the QR-si510 siRNA (10 nM) nanoplex for time period ranging from 12 hr to 1 week posttransfection. At the end of the incubation period, RNA was extracted, reverse transcribed and the LTR/RU5 gene expression quantitated using Q-PCR. Our results show a significant decrease in HIV-1 LTR gene expression in HIV-1-infected THP-1 cells treated with the 10 nM QR-si510 HIV-1siRNA nanoplex at 12, 24, 48, 96 hr, and 1 week posttransfection as compared to the untransfected THP-1 cells and the scrambled control. The results shown are mean ± SD of 3 separate experiments done in duplicate.

Mentions: Additional experiments to determine changes in viral replication by measuring HIV-1 LTR gene expression using real-time PCR were done using 10 nM concentration of the QR-si510 HIV-1 siRNA nanoplex at various time points ranging from 12 hr to 1 week posttransfection. The results of our HIV-1 LTR gene expression studies show a significant decrease in HIV-1 LTR gene expression in THP-1 cells treated with the 10 nM QR-si510 HIV-1siRNA nanoplex at 12, 24, 48, 96 hr, and 1 week posttransfection as compared to the untransfected THP-1 cells and also the scrambled control. The percentage decrease in HIV-1 LTR gene expression was 91% (TAI = 0.11 ± 0.02 versus 0.91 ± 0.05, (P < .0001)), 86% (TAI = 0.14 ± 0.06 versus 0.89 ± 0.03 (P < .0001)), 92% (TAI = 0.08 ± 0.03 versus 0.90 ± 0.07 (P < .0001)), 92% (TAI = 0.08 ± 0.05 versus 0.88 ± 0.03 (P < .0001)), and 91% (TAI = 0.09 ± 0.04 versus 0.94 ± 0.08 (P < .0001)) as compared to the scrambled control (Figure 5). The percentage decrease in HIV-1 LTR gene expression in THP-1 cells treated with the 10 nM of si510 HIV-1siRNA transfected with Lipofectamine at 12, 24, 48, 96 hr, and 1 week posttransfection were 60% (TAI = 0.40 ± 0.09 versus 0.91 ± 0.05, (P < .001)), 69% (TAI = 0.31 ± 0.07 versus 0.89 ± 0.03 (P < .001)), 77% (TAI = 0.23 ± 0.05 versus 0.90 ± 0.07 (P < .001)), 51% (TAI = 0.49 ± 0.11 versus 0.88 ± 0.03 (P < .004)), and 37% (TAI = 0.63 ± 0.06 versus 0.94 ± 0.08 (P < .006)), respectively, as compared to the scrambled control. These results confirm that the QR nanoparticles are a more effective than the commercially available transfection agent lipofectamine in not only delivering siRNA but also suppressing HIV-1 viral replication for a prolonged period of time.


Nanotherapeutics Using an HIV-1 Poly A and Transactivator of the HIV-1 LTR-(TAR-) Specific siRNA.

Mahajan SD, Aalinkeel R, Reynolds JL, Nair B, Sykes DE, Law WC, Ding H, Bergey EJ, Prasad PN, Schwartz SA - Patholog Res Int (2011)

Effect of QR-si510 siRNA nanoplex on LTR/RU5 gene expression in HIV-1-infected THP-1 cells. THP-1 monocytic cultures were treated with lipofectamine transfected scrambled siRNA (10 nM), Lipofectamine transfected si510 HIV-1 siRNA (10 nM) and  the QR-si510 siRNA (10 nM) nanoplex for time period ranging from 12 hr to 1 week posttransfection. At the end of the incubation period, RNA was extracted, reverse transcribed and the LTR/RU5 gene expression quantitated using Q-PCR. Our results show a significant decrease in HIV-1 LTR gene expression in HIV-1-infected THP-1 cells treated with the 10 nM QR-si510 HIV-1siRNA nanoplex at 12, 24, 48, 96 hr, and 1 week posttransfection as compared to the untransfected THP-1 cells and  the scrambled control.  The results shown are mean ± SD of 3 separate experiments done in duplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3108557&req=5

fig5: Effect of QR-si510 siRNA nanoplex on LTR/RU5 gene expression in HIV-1-infected THP-1 cells. THP-1 monocytic cultures were treated with lipofectamine transfected scrambled siRNA (10 nM), Lipofectamine transfected si510 HIV-1 siRNA (10 nM) and the QR-si510 siRNA (10 nM) nanoplex for time period ranging from 12 hr to 1 week posttransfection. At the end of the incubation period, RNA was extracted, reverse transcribed and the LTR/RU5 gene expression quantitated using Q-PCR. Our results show a significant decrease in HIV-1 LTR gene expression in HIV-1-infected THP-1 cells treated with the 10 nM QR-si510 HIV-1siRNA nanoplex at 12, 24, 48, 96 hr, and 1 week posttransfection as compared to the untransfected THP-1 cells and the scrambled control. The results shown are mean ± SD of 3 separate experiments done in duplicate.
Mentions: Additional experiments to determine changes in viral replication by measuring HIV-1 LTR gene expression using real-time PCR were done using 10 nM concentration of the QR-si510 HIV-1 siRNA nanoplex at various time points ranging from 12 hr to 1 week posttransfection. The results of our HIV-1 LTR gene expression studies show a significant decrease in HIV-1 LTR gene expression in THP-1 cells treated with the 10 nM QR-si510 HIV-1siRNA nanoplex at 12, 24, 48, 96 hr, and 1 week posttransfection as compared to the untransfected THP-1 cells and also the scrambled control. The percentage decrease in HIV-1 LTR gene expression was 91% (TAI = 0.11 ± 0.02 versus 0.91 ± 0.05, (P < .0001)), 86% (TAI = 0.14 ± 0.06 versus 0.89 ± 0.03 (P < .0001)), 92% (TAI = 0.08 ± 0.03 versus 0.90 ± 0.07 (P < .0001)), 92% (TAI = 0.08 ± 0.05 versus 0.88 ± 0.03 (P < .0001)), and 91% (TAI = 0.09 ± 0.04 versus 0.94 ± 0.08 (P < .0001)) as compared to the scrambled control (Figure 5). The percentage decrease in HIV-1 LTR gene expression in THP-1 cells treated with the 10 nM of si510 HIV-1siRNA transfected with Lipofectamine at 12, 24, 48, 96 hr, and 1 week posttransfection were 60% (TAI = 0.40 ± 0.09 versus 0.91 ± 0.05, (P < .001)), 69% (TAI = 0.31 ± 0.07 versus 0.89 ± 0.03 (P < .001)), 77% (TAI = 0.23 ± 0.05 versus 0.90 ± 0.07 (P < .001)), 51% (TAI = 0.49 ± 0.11 versus 0.88 ± 0.03 (P < .004)), and 37% (TAI = 0.63 ± 0.06 versus 0.94 ± 0.08 (P < .006)), respectively, as compared to the scrambled control. These results confirm that the QR nanoparticles are a more effective than the commercially available transfection agent lipofectamine in not only delivering siRNA but also suppressing HIV-1 viral replication for a prolonged period of time.

Bottom Line: We used a well-validated siRNA (si510) which targets the poly A/TAR (transactivator of the HIV-1 LTR) site and suppresses viral replication.Our results suggest that the QR-si510 HIV-1 siRNA nanoplex is not only effective in delivering siRNA, but also in suppressing HIV-1 viral replication for a longer time period.HIV-1 nanotherapeutics can thus enhance systemic bioavailability and offer multifunctionality.

View Article: PubMed Central - PubMed

Affiliation: Division of Allergy, Immunology, and Rheumatology, Department of Medicine, University at Buffalo, The State University of New York, 640 Ellicott Street, Room 444 Innovation Center, Buffalo, NY 14203, USA.

ABSTRACT
HIV-1 replication can be efficiently inhibited by intracellular expression of an siRNA targeting the viral RNA. We used a well-validated siRNA (si510) which targets the poly A/TAR (transactivator of the HIV-1 LTR) site and suppresses viral replication. Nanotechnology holds much potential for impact in the field of HIV-1 therapeutics, and nanoparticles such as quantum rods (QRs) can be easily functionalized to incorporate siRNA forming stable nanoplexes that can be used for gene silencing. We evaluated the efficacy of the QR-si510 HIV-1 siRNA nanoplex in suppressing viral replication in the HIV-1-infected monocytic cell line THP-1 by measuring p24 antigen levels and gene expression levels of HIV-1 LTR. Our results suggest that the QR-si510 HIV-1 siRNA nanoplex is not only effective in delivering siRNA, but also in suppressing HIV-1 viral replication for a longer time period. HIV-1 nanotherapeutics can thus enhance systemic bioavailability and offer multifunctionality.

No MeSH data available.


Related in: MedlinePlus