Establishment of a high-throughput screening system for universal anti-HIV targets
详细信息 查看全文
- 作者：Qi Yin (1) (2)
DaoMing Zhuang (3)
YaQin Jiang (1) (2)
ChuanKe Zhao (1) (2)
Xin Zeng (1) (2)
ShiYou Li (1) - 关键词：HIV ; 1 ; high ; throughput screening ; inhibitor ; virus ; cell ; based assay
- 刊名：Chinese Science Bulletin
- 出版年：2010
- 出版时间：April 2010
- 年：2010
- 卷：55
- 期：10
- 页码：937-942
- 全文大小：754KB
- 参考文献：1. Hartzell J D, Janke I E, Weintrob A C. Impact of depression on HIV outcomes in the HAART era. J Antimicrob Chemother, 2008, 62: 246-55 CrossRef
2. Nasoodi A, Lim L T, Al-Ani A, et al. What you can see in your patient’s eyes? Review of ocular manifestations of HIV in HAART era. Intl J STD AIDS, 2008, 19: 4-1 CrossRef
3. Aquaro S, Svicher V, Ronga L, et al. HIV-1-associated dementia during HAART therapy. Recent patents on CNS drug discovery, 2008, 3: 23-3 CrossRef
4. Martins S, Ramos M J, Fernandes P A. The current status of the NNRTI family of antiretrovirals used in the HAART regime against HIV infection. Curr Med Chem, 2008, 15: 1083-095 CrossRef
5. Schurmann D, Wesselmann H, Kurowski M, et al. Kaletra single agent HAART after intolerance of NRTI- and NNRTI-containing regimens following kidney transplantation. Infection, 2007, 35: 194-96 CrossRef
6. Bongiovanni M, Gianotti N, Chiesa E, et al. Observational study on HIV-infected subjects failing HAART receiving tenofovir plus didanosine as NRTI backbone. Infection, 2007, 35: 451-56 CrossRef
7. Apuzzo L G, Vaida F, Gallant J E, et al. Tolerability and efficacy of PI versus NNRTI-based regimens in subjects receiving HAART during acute or early HIV infection. J Acquir Immune Defic Syndr, 2009, 50: 267-75 CrossRef
8. Young J, Rickenbach M, Weber R, et al. Body fat changes among antiretroviral-naive patients on PI- and NNRTI-based HAART in the Swiss HIV cohort study. Antivir Ther, 2005, 10: 73-1
9. Shafer R W, Schapiro J M. HIV-1 drug resistance mutations: An updated framework for the second decade of HAART. AIDS Rev, 2008, 10: 67-4
10. Yeni P. Update on HAART in HIV. J Hepatol, 2006, 44: S100–S103 CrossRef
11. Kuritzkes D R. HIV-1 entry inhibitors: An overview. Curr Opin HIV AIDS, 2009, 4: 82-7 CrossRef
12. Qian K, Morris-Natschke S L, Lee K H. HIV entry inhibitors and their potential in HIV therapy. Med Res Rev, 2009, 29: 369-93 CrossRef
13. Al-Mawsawi L Q, Al-Safi R I, Neamati N. Anti-infectives: Clinical progress of HIV-1 integrase inhibitors. Expert Opin Emerg Drugs, 2008, 13: 213-25 CrossRef
14. Engelman A. Mechanistic and pharmacological analyses of HIV-1 integration. Methods, 2009, 47: 225-28 CrossRef
15. Goffinet C, Allespach I, Oberbremer L, et al. Pharmacovirological impact of an integrase inhibitor on HIV-1 cDNA Species / in vivo. J Virol, 2009, doi:10.1128/JVI.00683-09
16. Mehta N, Reilly M. Atherosclerotic cardiovascular disease risk in the HAART-treated HIV-1 population. HIV Clin Trials, 2005, 6: 5-4 CrossRef
17. Sung H, Jung Y S, Kang M W, et al. High frequency of drug resistance mutations in human immunodeficiency virus type 1-infected Korean patients treated with HAART. AIDS Res Human Retrovir, 2007, 23: 1223-229 CrossRef
18. Wang Y M, Dyer W B, Workman C, et al. Drug resistance and viral evolution in plasma and peripheral blood cells during structured treatment interruption (STI) and non-interrupted HAART. Curr HIV Res, 2007, 5: 235-50 CrossRef
19. Chew C B, Potter S J, Wang B, et al. Assessment of drug resistance mutations in plasma and peripheral blood mononuclear cells at different plasma viral loads in patients receiving HAART. J Clin Virol, 2005, 33: 206-16
20. Garcia J M, Gao A, He P L, et al. High-throughput screening using pseudotyped lentiviral particles: A strategy for the identification of HIV-1 inhibitors in a cell-based assay. Antiv Res, 2009, 81: 239-47 CrossRef
21. Kempf M C, Jones J, Heil M L, et al. A high-throughput drug screening system for HIV-1 transcription inhibitors. J Biomol Screen, 2006, 11: 807-15 CrossRef
22. Ferraz R M, Aris A, Martinez M A, et al. High-throughput, functional screening of the anti-HIV-1 humoral response by an enzymatic nanosensor. Mol Immunol, 2006, 43: 2119-123 CrossRef
23. John S, Fletcher T M, Jonsson C B. Development and application of a high-throughput screening assay for HIV-1 integrase enzyme activities. J Biomol Screen, 2005, 10: 606-14 CrossRef
24. Fakes M G, Vakkalagadda B J, Qian F, et al. Enhancement of oral bioavailability of an HIV-attachment inhibitor by nanosizing and amorphous formulation approaches. Intl J Pharm, 2009, 370: 167-74 CrossRef
25. Wen Z, Stern S T, Martin D E, et al. Structural characterization of anti-HIVdrug candidate PA-457 [3-O-(3-3-dimethylsuccinyl)-betulinic acid] and its acyl glucuronides in rat bile and evaluation of in vitro stability in human and animal liver microsomes and plasma. Drug Metab Dispos, 2006, 34: 1436-442 CrossRef
26. Cao J, Isaacson J, Patick A K, et al. High-throughput human immunodeficiency virus type 1 (HIV-1) full replication assay that includes HIV-1 Vif as an antiviral target. Antimicrob Agents Chemother, 2005, 49: 38333841
27. Van Duyne R, Easley R, Wu W, et al. Lysine methylation of HIV-1 Tat regulates transcriptional activity of the viral LTR. Retrovirology, 2008, 5: 40 CrossRef
28. Dejucq N. HIV-1 replication in CD4+ T cell lines: The effects of adaptation on co-receptor use, tropism, and accessory gene function. J Leukocyte Biol, 2000, 68: 331-37
29. Barin F, Brunet S, Brand D, et al. Interclade neutralization and enhancement of human immunodeficiency virus type 1 identified by an assay using HeLa cells expressing both CD4 receptor and CXCR4/CCR5 coreceptors. J Infect Dis, 2004, 189: 322-27 CrossRef
30. Pauwels R, Andries K, Desmyter J, et al. Potent and selective inhibittion of HIV-1 replication in vitro by a novel series of TIBO derivatives. Nature, 1990, 343: 470-74 CrossRef
31. Wild C, Oas T, McDanal C, et al. A synthetic peptide inhibitor of human immunodeficiency virus replication: Correlation between solution structure and viral inhibition. Proc Natl Acad Sci USA, 1992, 89: 10537-0541 CrossRef
32. Squires K E. An introduction to nucleoside and nucleotide analogues. Antivir Ther, 2001, 6(Suppl 3): 1-4
33. Adelson M E, Pacchia A L, Kaul M, et al. Toward the development of a virus-cell-based assay for the discovery of novel compounds against human immunodeficiency virus type 1. Antimicrob Agents Chemoth (Bethesda), 2003, 47: 501-08 CrossRef
34. Miyake H, Iizawa Y, Baba M. Novel reporter T-cell line highly susceptible to both CCR5- and CXCR4-using human immunodeficiency virus type 1 and its application to drug susceptibility tests. J Clin Microbiol, 2003, 41: 2515-521 CrossRef
35. Blair W S, Isaacson J, Li X, et al. A novel HIV-1 antiviral high throughput screening approach for the discovery of HIV-1 inhibitors. Antivir Res, 2005, 65: 107-16 CrossRef - 作者单位：Qi Yin (1) (2)
DaoMing Zhuang (3)
YaQin Jiang (1) (2)
ChuanKe Zhao (1) (2)
Xin Zeng (1) (2)
ShiYou Li (1)
1. Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100029, China
2. Graduate University of the Chinese Academy of Sciences, Beijing, 100049, China
3. State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing, 100071, China - ISSN：1861-9541
文摘
The process of identifying novel human immunodeficiency virus 1 (HIV-1) inhibitors presents a challenge for industrial and scientific research. Virus-cell-based screening approaches offer some advantages in the quest for novel inhibitors because they include multiple targets in a single screen and in some cases reveal targets not captured in biochemical assays. In this study, a high-throughput screening (HTS) system for HIV-1 inhibitors was developed, which allows the simultaneous screening of all the HIV-1 targets required for replication in the cell culture. HeLa/cluster of differentiation 4 (CD4)/long terminal repeat (LTR) indicator cells, which stably expressed high levels of HIV receptor CD4 and contained the firefly luciferase reporter gene under the control of the HIV-1 LTR promoter, were generated. The expression of CD4 and LTR function in this cell line was validated by Western blot and luciferase assay. MT2 cells, a human T-cell leukemia cell line that support high levels of HIV-1 replication, were infected with HIV-1, and then the infected MT2 cells were co-cultured with HeLa/CD4/LTR cells. In the optimized assay conditions, HIV-1 replication occurs rapidly in the MT2 cells, resulting in the infection of the HeLa/CD4/LTR cells and a significant induction of luciferase signals through LTR, which is activated by the expression of HIV-1 tat gene. The luciferase signals of HeLa/CD4/LTR cells co-cultured with HIV-1 infected MT2 cells were significantly stronger than the signals of noninfected HeLa/CD4/LTR cells (P < 0.001). The inhibitory effects of HIV-1 inhibitors (3-Azido-3-deoxythymidine [AZT], efavirenz [EFV], and nevirapine [NVP]) were evaluated with this assay, and the inhibitory concentration 50% (IC50) values of the above three inhibitors were 58, 1.4, and 85 nmol/L, respectively, indicating that the assay provides the necessary sensitivity for identifying antiviral molecules. The Z-factor had a value of 0.563, indicating this is a very robust assay. These results suggested that HIV-1 infection assay represents a novel approach to HIV-1 antiviral screening that allows for the effective execution of HTS campaigns.