Effects of CC Chemokine Receptor 5 (CCR5) Inhibitors on the Dynamics of CCR5 and CC-chemokine-CCR5 Interactions

Overview

Journal Antivir Ther

Publisher Sage Publications

Specialties Infectious Diseases
Microbiology

Date 2010 Jun 3

PMID 20516552

Citations 3

Authors

Hirotomo Nakata

Michael Kruhlak

Wakako Kamata

Hiromi Ogata-Aoki

Jianfeng Li

Kenji Maeda

Arun K Ghosh

Hiroaki Mitsuya

Affiliations

Soon will be listed here.

Abstract

Background: This study aimed to examine how CC chemokine receptor 5 (CCR5) inhibitors (aplaviroc [APL], TAK779 and maraviroc [MVC]) interact with CCR5 and affect its dynamics and physiological CC-chemokine-CCR5 interactions.

Methods: A yellow fluorescent protein (YFP)-tagged CCR5-expressing U373-MAGI cell line was generated and a stable CCR5-expressing clonal population, (YFP)CCR5-UM16, was prepared. (YFP)CCR5-UM16 cells were exposed to RANTES, macrophage inflammatory protein (MIP)-1alpha or MIP-1beta (all 100 ng/ml) with or without CCR5 inhibitors and (YFP)CCR5 internalization was visualized with real-time by laser scanning confocal microscopy. The mobility of (YFP)CCR5 was also examined in the presence of CCR5 inhibitors with fluorescence recovery after photobleaching (FRAP) imaging.

Results: Following the addition of each CC chemokine, intracellular fluorescence intensity increased whereas membranous fluorescence decreased, signifying (YFP)CCR5 internalization. All three CCR5 inhibitors failed to induce (YFP)CCR5 internalization. All three CCR5 inhibitors blocked the CC-chemokine-induced internalization at a high concentration of 1 microM; however, the ratio of APL concentration that blocked RANTES-induced internalization by 50% over APL concentration that blocked HIV type-1 (HIV-1) replication by 50% was 16.4, indicating that APL permits CC-chemokine-induced internalization to a much greater extent compared with TAK779 and MVC, having ratios of 1.1 and 0.9, respectively. The examination of (YFP)CCR5 mobility with FRAP imaging revealed that (YFP)CCR5 continuously underwent rapid redistribution, which none of the three inhibitors blocked.

Conclusions: The finding that APL moderately blocked the RANTES-triggered (YFP)CCR5 internalization despite the highly potent anti-HIV-1 activity of APL strongly suggests that development of CCR5 inhibitors, which do not overly inhibit physiological CC-chemokine-CCR5 interactions, is practically feasible.

Citing Articles

Activity and structural analysis of GRL-117C: a novel small molecule CCR5 inhibitor active against R5-tropic HIV-1s.

Nakata H, Maeda K, Das D, Chang S, Matsuda K, Venkateswara Rao K Sci Rep. 2019; 9(1):4828.

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Immunological and pharmacological strategies to reactivate HIV-1 from latently infected cells: a possibility for HIV-1 paediatric patients?.

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The immunologic effects of maraviroc intensification in treated HIV-infected individuals with incomplete CD4+ T-cell recovery: a randomized trial.

Hunt P, Shulman N, Hayes T, Dahl V, Somsouk M, Funderburg N Blood. 2013; 121(23):4635-46.

PMID: 23589670 PMC: 3685899. DOI: 10.1182/blood-2012-06-436345.

References

Makino Y, Cook D, Smithies O, Hwang O, Neilson E, Turka L . Impaired T cell function in RANTES-deficient mice. Clin Immunol. 2002; 102(3):302-9. DOI: 10.1006/clim.2001.5178. View

Alkhatib G, Combadiere C, Broder C, Feng Y, Kennedy P, Murphy P . CC CKR5: a RANTES, MIP-1alpha, MIP-1beta receptor as a fusion cofactor for macrophage-tropic HIV-1. Science. 1996; 272(5270):1955-8. DOI: 10.1126/science.272.5270.1955. View

Maeda K, Das D, Ogata-Aoki H, Nakata H, Miyakawa T, Tojo Y . Structural and molecular interactions of CCR5 inhibitors with CCR5. J Biol Chem. 2006; 281(18):12688-98. DOI: 10.1074/jbc.M512688200. View

Baba M, Nishimura O, Kanzaki N, Okamoto M, Sawada H, Iizawa Y . A small-molecule, nonpeptide CCR5 antagonist with highly potent and selective anti-HIV-1 activity. Proc Natl Acad Sci U S A. 1999; 96(10):5698-703. PMC: 21923. DOI: 10.1073/pnas.96.10.5698. View

Raport C, Gosling J, Schweickart V, Gray P, Charo I . Molecular cloning and functional characterization of a novel human CC chemokine receptor (CCR5) for RANTES, MIP-1beta, and MIP-1alpha. J Biol Chem. 1996; 271(29):17161-6. DOI: 10.1074/jbc.271.29.17161. View

Eri R, Jonsson J, Pandeya N, Purdie D, Clouston A, Martin N . CCR5-Delta32 mutation is strongly associated with primary sclerosing cholangitis. Genes Immun. 2004; 5(6):444-50. DOI: 10.1038/sj.gene.6364113. View

Wysocki C, Jiang Q, Panoskaltsis-Mortari A, Taylor P, McKinnon K, Su L . Critical role for CCR5 in the function of donor CD4+CD25+ regulatory T cells during acute graft-versus-host disease. Blood. 2005; 106(9):3300-7. PMC: 1895335. DOI: 10.1182/blood-2005-04-1632. View

Martinson J, Chapman N, Rees D, Liu Y, Clegg J . Global distribution of the CCR5 gene 32-basepair deletion. Nat Genet. 1997; 16(1):100-3. DOI: 10.1038/ng0597-100. View

Gulick R, Lalezari J, Goodrich J, Clumeck N, DeJesus E, Horban A . Maraviroc for previously treated patients with R5 HIV-1 infection. N Engl J Med. 2008; 359(14):1429-41. PMC: 3078519. DOI: 10.1056/NEJMoa0803152. View

10.

Fatkenheuer G, Pozniak A, Johnson M, Plettenberg A, Staszewski S, Hoepelman A . Efficacy of short-term monotherapy with maraviroc, a new CCR5 antagonist, in patients infected with HIV-1. Nat Med. 2005; 11(11):1170-2. DOI: 10.1038/nm1319. View

11.

Maeda K, Nakata H, Koh Y, Miyakawa T, Ogata H, Takaoka Y . Spirodiketopiperazine-based CCR5 inhibitor which preserves CC-chemokine/CCR5 interactions and exerts potent activity against R5 human immunodeficiency virus type 1 in vitro. J Virol. 2004; 78(16):8654-62. PMC: 479103. DOI: 10.1128/JVI.78.16.8654-8662.2004. View

12.

Maeda K, Yoshimura K, Shibayama S, Habashita H, Tada H, Sagawa K . Novel low molecular weight spirodiketopiperazine derivatives potently inhibit R5 HIV-1 infection through their antagonistic effects on CCR5. J Biol Chem. 2001; 276(37):35194-200. DOI: 10.1074/jbc.M105670200. View

13.

Maeda K, Das D, Yin P, Tsuchiya K, Ogata-Aoki H, Nakata H . Involvement of the second extracellular loop and transmembrane residues of CCR5 in inhibitor binding and HIV-1 fusion: insights into the mechanism of allosteric inhibition. J Mol Biol. 2008; 381(4):956-74. PMC: 2630503. DOI: 10.1016/j.jmb.2008.06.041. View

14.

Lederman M, Penn-Nicholson A, Cho M, Mosier D . Biology of CCR5 and its role in HIV infection and treatment. JAMA. 2006; 296(7):815-26. DOI: 10.1001/jama.296.7.815. View

15.

Steffens C, Hope T . Mobility of the human immunodeficiency virus (HIV) receptor CD4 and coreceptor CCR5 in living cells: implications for HIV fusion and entry events. J Virol. 2004; 78(17):9573-8. PMC: 506925. DOI: 10.1128/JVI.78.17.9573-9578.2004. View

16.

Kruhlak M, Crouch E, Orlov M, Montano C, Gorski S, Nussenzweig A . The ATM repair pathway inhibits RNA polymerase I transcription in response to chromosome breaks. Nature. 2007; 447(7145):730-4. DOI: 10.1038/nature05842. View

17.

Longden J, Cooke E, Hill S . Effect of CCR5 receptor antagonists on endocytosis of the human CCR5 receptor in CHO-K1 cells. Br J Pharmacol. 2008; 153(7):1513-27. PMC: 2437898. DOI: 10.1038/sj.bjp.0707691. View

18.

Huang Y, Paxton W, Wolinsky S, Neumann A, Zhang L, He T . The role of a mutant CCR5 allele in HIV-1 transmission and disease progression. Nat Med. 1996; 2(11):1240-3. DOI: 10.1038/nm1196-1240. View

19.

Woitas R, Ahlenstiel G, Iwan A, Rockstroh J, Brackmann H, Kupfer B . Frequency of the HIV-protective CC chemokine receptor 5-Delta32/Delta32 genotype is increased in hepatitis C. Gastroenterology. 2002; 122(7):1721-8. DOI: 10.1053/gast.2002.33660. View

20.

Kiss D, Longden J, Fechner G, Avery V . The functional antagonist Met-RANTES: a modified agonist that induces differential CCR5 trafficking. Cell Mol Biol Lett. 2009; 14(4):537-47. PMC: 6275935. DOI: 10.2478/s11658-009-0017-1. View