Lentiviral transduction of Tar Decoy and CCR5 ribozyme into CD34+ progenitor cells and derivation of HIV-1 resistant T cells and macrophages

AIDS Research and Therapy

Akhil Banerjea¹  Ming-Jie Li²  Leila Remling¹  John Rossi²  Ramesh Akkina¹ 

[1] Dept. Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado 80523, USA

[2] Division of Molecular Biology, Beckman Research Institute of the City of Hope, 1450 East Duarte Road, Duarte, California, 91010, USA

关键词: CD34 cells;    HIV aptamers;    Lentiviral vectors;    SCID-hu mice;    CCR5 ribozyme;    HIV tar decoy;    AIDS gene therapy;   

Others  :  790118
DOI  :  10.1186/1742-6405-1-2

 received in 2004-10-01, accepted in 2004-12-17,  发布年份 2004

【 摘 要 】

Background

RNA based antiviral approaches against HIV-1 are among the most promising for long-term gene therapy. These include ribozymes, aptamers (decoys), and small interfering RNAs (siRNAs). Lentiviral vectors are ideal for transduction of such inhibitory RNAs into hematopoietic stem cells due to their ability to transduce non-dividing cells and their relative refractiveness to gene silencing. The objective of this study is to introduce an HIV-1 Tar aptamer either alone or in combination with an anti-CCR5 ribozyme into CD34+ hematopoietic progenitor cells via an HIV-based lentiviral vector to derive viral resistant progeny T cells and macrophages.

Results

High efficiency and sustained gene transfer into CD34+ cells were achieved with lentiviral vector constructs harboring either Tar decoy or Tar decoy in combination with CCR5 ribozyme. Cells transduced with these constructs differentiated normally into T-lymphocytes in vivo in thy/liv grafts of SCID-hu mice, and into macrophages in vitro in the presence of appropriate growth factors. When challenged in vitro, the differentiated T lymphocytes and macrophages showed marked resistance against HIV-1 infection.

Conclusions

Viral resistant transgenic T cells and macrophages that express HIV-1 Tar aptamer either alone or in combination with an anti-CCR5 ribozyme could be obtained by lentiviral gene transduction of CD34+ progenitor cells. These results showed for the first time that expression of these anti-HIV-1 transgenes in combination do not interfere with normal thymopoiesis and thus have set the stage for their application in stem cell based gene therapy for HIV/AIDS.

【 授权许可】

   
2004 Banerjea et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

Figure 8.

【 参考文献 】

• [1]Stevenson M: HIV-1 Pathogenesis. Nat Med 2003, 9:853-860.
• [2]Berger EA, Murphy PM, Farber JM: Chemokine receptors as HIV-1 coreceptors: Roles in viral entry, tropism, and disease. Annu Rev Immunol 1999, 17:657-700.
• [3]Liu R, Paxton WA, Choe S, Ceradini D, Martin SR, Horuk R, MacDonald ME, Stuhlmann H, Koup RA, Landau NR: Homozygous defect in HIV-1 coreceptor accounts for resistance of some multiply exposed individuals to HIV-1 infection. Cell 1996, 86:367-377.
• [4]Goila R, Banerjea AC: Sequence specific cleavage of HIV-1 coreceptor-CCR5 gene by a hammer-head ribozyme and a DNA-enzyme. Inhibition of the coreceptor function by DNA-enzymes. FEBS Lett 1998, 436:233-238.
• [5]Bai J, Rossi J, Akkina R: Multivalent anti-CCR5 ribozymes for stem cell-based HIV type 1 gene therapy. AIDS Res Hum Retroviruses 2001, 17:385-399.
• [6]Bai J, Gorantla S, Banda N, Cagnon L, Rossi J, Akkina R: Characterization of anti-CCR5 ribozyme transduced CD34+ hematopoetic progenitor cells in vitro and in a SCID-hu mouse model in vivo. Mol Ther 2000, 1:244-254.
• [7]Qin XF, An DS, Chen IS, Baltimore D: Inhibiting HIV-1 infection in human T cells by lentiviral-mediated delivery of small interfering RNA against CCR5. Proc Natl Acad Sci USA 2003, 100:183-188.
• [8]Anderson J, Banerjea A, Akkina R: Bispecific short hairpin siRNA constructs targeted to CD4, CXCR4, and CCR5 confer HIV-1 resistance. Oligonucleotides 2003, 13:303-312.
• [9]Lisziewicz J, Sun D, Smythe J, Lusso P, Lori F, Louie A, Markham P, Rossi J, Reitz M, Gallo RC: Inhibition of human immunodeficiency virus type 1 by regulated expression of polymeric TAT activation response RNA decoy as a strategy for gene therapy for AIDS. Proc Natl Acad Sci USA 1993, 90:8000-8004.
• [10]Michienzi A, Li S, Zaia JA, Rossi JJ: A Nucleolar TAR decoy inhibitor of HIV-1 replication. Proc Natl Acad Sci USA 2002, 99:14047-14052.
• [11]Rossi J: The application of ribozyme to HIV-1 infection. Curr Opin Mol Ther 1999, 1:316-322.
• [12]Coburn GA, Cullen BR: Potent and specific inhibition of human immunodeficiency virus type 1 replication by RNA interference. J Virology 2002, 76:9225-9231.
• [13]Lee NS, Dohjima T, Bauer G, Li H, Li MJ, Ehsani A, Salvaterra P, Rossi J: Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells. Nat Biotechnol 2002, 19:500-505.
• [14]Boden D, Pusch O, Lee F, Tucker L, Ramratnam B: Human immunodeficiency virus type 1 escape from RNA interference. J Virology 2003, 77:11531-11535.
• [15]Kohn DB, Bauer G, Rice CR, Rothschild JC, Carbonaro DA, Valdez P, Hao Q, Zhou C, Bahner I, Kearns K, Brody K, Fox S, Haden E, Wilson K, Salata C, Dolan C, Wetter C, Aguilar-Cordova E, Church J: A clinical trial of retroviral mediated transfer of a rev-response element decoy gene into CD34+ cells from the bone marrow of human immunodeficiency virus -1 infected children. Blood 1999, 94:368-371.
• [16]Engel BC, Kohn BD: Stem cell directed gene therapy. Front Biosci 1999, 4:E26-33.
• [17]Robbins PB, Skelton DC, Yu XJ, Halene S, Leonard EH, Kohn DB: Consistent, persistent expression from modified retroviral vectors in murine hematopoietic stem cells. Proc Natl Acad Sci USA 1998, 95:10182-10187.
• [18]Li MJ, Bauer G, Michienzi A, Yee JK, Lee NS, Kim J, Li S, Castanotto D, Zaia J, Rossi JJ: Inhibition of HIV-1 infection by lentiviral vectors expressing Pol III-promoted anti-HIV RNAs. Mol Ther 2003, 8:196-206.
• [19]Cagnon L, Rossi JJ: Downregulation of the CCR5 beta-chemokine receptor and inhibition of HIV-1 inhibition by stable VA1 -ribozyme chimeric transcripts. Antisense Nucleic Acid Drug Dev 2000, 10:251-161.
• [20]Bai J, Banda N, Lee NS, Rossi J, Akkina R: RNA-based anti-HIV-1 gene therapeutic constructs in SCID-hu mouse model. Mol Ther 2002, 6:770-782.
• [21]Yam PY, Li S, Wu J, Hu J, Zaia JA, Yee JK: Design of HIV-1 vectors for efficient gene delivery into human hematopoietic cells. Mol Ther 2002, 5:479-484.
• [22]Banda NK, Akkina R, Terrell K, Shpall EJ, Tomczak J, Campain J, Claman H, Cagle L, Harrison GS: Diptheria toxin A gene -mediated HIV-1 protection of cord blood-derived T cells in the SCID-hu mouse model. J Hematother 1998, 7:319-331.
• [23]Banerjea AC, Li MJ, Bauer G, Remling L, Lee NS, Rossi J, Akkina R: Inhibition of HIV-1 by lentiviral vector-transduced siRNAs in T lymphocytes differentiated in SCID-hu mice and CD34+ progenitor cell-derived macrophages. Mol Ther 2003, 8:62-71.
• [24]Germain RN: T-cell development and the CD4-CD8 lineage decision. Nat Rev Immunol 2002, 2:309-322.
• [25]Akkina R, Walton RM, Chen ML, Li QX, Planelles V, Chen IS: High efficiency gene transfer into CD34+ cells with a HIV-1 based retroviral vector pseudotyped with VSV-G. J Virology 1996, 70:2581-2585.
• [26]Akkina R, Rosenblatt JD, Campbell AG, Chen IS, Zack JA: Modeling human lymphoid precursor cell gene therapy in SCID-hu mouse. Blood 1994, 84:1393-1398.