Transduction of human CD34+ hematopoietic progenitor cells by a retroviral vector expressing an RRE decoy inhibits human immunodeficiency virus type 1 replication in myelomonocytic cells produced in long-term culture (original) (raw)

Abstract

Genetic modification of hematopoietic stem cells with a synthetic "anti-human immunodeficiency virus type 1 (HIV-1) gene" which inhibits replication of HIV-1 may allow production of mature lymphoid and monocytic cells resistant to HIV-1 growth after autologous transplantation. Because productive HIV-1 replication requires binding of the Rev protein to the Rev-responsive element (RRE) within the viral transcripts for the HIV-1 structural proteins, anti-HIV-1 gene products which interfere with Rev-RRE interactions may inhibit HIV-1 replication. One such strategy involves overexpression of the RRE sequences in transcripts derived from retroviral vectors to act as decoys to sequester Rev protein and prevent its binding to the RRE element in HIV-1 transcripts. We developed an in vitro model to test the efficacy of this gene therapy approach in primary human hematopoietic cells. Human CD34+ hematopoietic progenitor cells from normal bone marrow or umbilical cord blood were transduced with retroviral vectors carrying RRE decoy sequences as part of a long terminal repeat-directed transcript expressing the neo gene (L-RRE-neo) or with a control vector expressing only the neo gene (LN). The transduced progenitors were allowed to differentiate into mature myelomonocytic cells which were able to support vigorous growth of the monocytotropic isolate of HIV-1, JR-FL. HIV-1 replication was measured in unselected cell populations and following G418 selection to obtain uniformly transduced cell populations. Inhibition of HIV-1 replication in the unselected cell cultures was between 50.2 and 76.7% and was highly effective (99.4 to 99.9%) in the G418-selected cultures. Progenitors transduced by either the L-RRE-neo vector or the control LN vector were identical with respect to hematopoietic growth and differentiation. These findings demonstrate the ability of an RRE decoy strategy to inhibit HIV-1 replication in primary human myelomonocytic cells after transduction of CD34+ progenitor cells, without adverse effects on hematopoietic cell function.

Full Text

The Full Text of this article is available as a PDF (321.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bahner I., Zhou C., Yu X. J., Hao Q. L., Guatelli J. C., Kohn D. B. Comparison of trans-dominant inhibitory mutant human immunodeficiency virus type 1 genes expressed by retroviral vectors in human T lymphocytes. J Virol. 1993 Jun;67(6):3199–3207. doi: 10.1128/jvi.67.6.3199-3207.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bevec D., Dobrovnik M., Hauber J., Böhnlein E. Inhibition of human immunodeficiency virus type 1 replication in human T cells by retroviral-mediated gene transfer of a dominant-negative Rev trans-activator. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9870–9874. doi: 10.1073/pnas.89.20.9870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brenner M. K., Rill D. R., Holladay M. S., Heslop H. E., Moen R. C., Buschle M., Krance R. A., Santana V. M., Anderson W. F., Ihle J. N. Gene marking to determine whether autologous marrow infusion restores long-term haemopoiesis in cancer patients. Lancet. 1993 Nov 6;342(8880):1134–1137. doi: 10.1016/0140-6736(93)92122-a. [DOI] [PubMed] [Google Scholar]
  4. Collins L. S., Dorshkind K. A stromal cell line from myeloid long-term bone marrow cultures can support myelopoiesis and B lymphopoiesis. J Immunol. 1987 Feb 15;138(4):1082–1087. [PubMed] [Google Scholar]
  5. Cullen B. R. Regulation of HIV-1 gene expression. FASEB J. 1991 Jul;5(10):2361–2368. doi: 10.1096/fasebj.5.10.1712325. [DOI] [PubMed] [Google Scholar]
  6. Curiel T. J., Cook D. R., Wang Y., Hahn B. H., Ghosh S. K., Harrison G. S. Long-term inhibition of clinical and laboratory human immunodeficiency virus strains in human T-cell lines containing an HIV-regulated diphtheria toxin A chain gene. Hum Gene Ther. 1993 Dec;4(6):741–747. doi: 10.1089/hum.1993.4.6-741. [DOI] [PubMed] [Google Scholar]
  7. Duan L., Bagasra O., Laughlin M. A., Oakes J. W., Pomerantz R. J. Potent inhibition of human immunodeficiency virus type 1 replication by an intracellular anti-Rev single-chain antibody. Proc Natl Acad Sci U S A. 1994 May 24;91(11):5075–5079. doi: 10.1073/pnas.91.11.5075. [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]
  8. Dunbar C. E., Cottler-Fox M., O'Shaughnessy J. A., Doren S., Carter C., Berenson R., Brown S., Moen R. C., Greenblatt J., Stewart F. M. Retrovirally marked CD34-enriched peripheral blood and bone marrow cells contribute to long-term engraftment after autologous transplantation. Blood. 1995 Jun 1;85(11):3048–3057. [PubMed] [Google Scholar]
  9. Gilboa E., Smith C. Gene therapy for infectious diseases: the AIDS model. Trends Genet. 1994 Apr;10(4):139–144. doi: 10.1016/0168-9525(94)90216-x. [DOI] [PubMed] [Google Scholar]
  10. Heaphy S., Finch J. T., Gait M. J., Karn J., Singh M. Human immunodeficiency virus type 1 regulator of virion expression, rev, forms nucleoprotein filaments after binding to a purine-rich "bubble" located within the rev-responsive region of viral mRNAs. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7366–7370. doi: 10.1073/pnas.88.16.7366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Joshi S., Van Brunschot A., Asad S., van der Elst I., Read S. E., Bernstein A. Inhibition of human immunodeficiency virus type 1 multiplication by antisense and sense RNA expression. J Virol. 1991 Oct;65(10):5524–5530. doi: 10.1128/jvi.65.10.5524-5530.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kohn D. B., Weinberg K. I., Nolta J. A., Heiss L. N., Lenarsky C., Crooks G. M., Hanley M. E., Annett G., Brooks J. S., el-Khoureiy A. Engraftment of gene-modified umbilical cord blood cells in neonates with adenosine deaminase deficiency. Nat Med. 1995 Oct;1(10):1017–1023. doi: 10.1038/nm1095-1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Koyanagi Y., Miles S., Mitsuyasu R. T., Merrill J. E., Vinters H. V., Chen I. S. Dual infection of the central nervous system by AIDS viruses with distinct cellular tropisms. Science. 1987 May 15;236(4803):819–822. doi: 10.1126/science.3646751. [DOI] [PubMed] [Google Scholar]
  14. Larder B. A., Kemp S. D. Multiple mutations in HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT). Science. 1989 Dec 1;246(4934):1155–1158. doi: 10.1126/science.2479983. [DOI] [PubMed] [Google Scholar]
  15. Lee S. W., Gallardo H. F., Gilboa E., Smith C. Inhibition of human immunodeficiency virus type 1 in human T cells by a potent Rev response element decoy consisting of the 13-nucleotide minimal Rev-binding domain. J Virol. 1994 Dec;68(12):8254–8264. doi: 10.1128/jvi.68.12.8254-8264.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lee T. C., Sullenger B. A., Gallardo H. F., Ungers G. E., Gilboa E. Overexpression of RRE-derived sequences inhibits HIV-1 replication in CEM cells. New Biol. 1992 Jan;4(1):66–74. [PubMed] [Google Scholar]
  17. Malim M. H., Freimuth W. W., Liu J., Boyle T. J., Lyerly H. K., Cullen B. R., Nabel G. J. Stable expression of transdominant Rev protein in human T cells inhibits human immunodeficiency virus replication. J Exp Med. 1992 Oct 1;176(4):1197–1201. doi: 10.1084/jem.176.4.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Markowitz M., Mo H., Kempf D. J., Norbeck D. W., Bhat T. N., Erickson J. W., Ho D. D. Selection and analysis of human immunodeficiency virus type 1 variants with increased resistance to ABT-538, a novel protease inhibitor. J Virol. 1995 Feb;69(2):701–706. doi: 10.1128/jvi.69.2.701-706.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miller A. D., Buttimore C. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol Cell Biol. 1986 Aug;6(8):2895–2902. doi: 10.1128/mcb.6.8.2895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miller A. D., Rosman G. J. Improved retroviral vectors for gene transfer and expression. Biotechniques. 1989 Oct;7(9):980-2, 984-6, 989-90. [PMC free article] [PubMed] [Google Scholar]
  21. Miller A. D., Trauber D. R., Buttimore C. Factors involved in production of helper virus-free retrovirus vectors. Somat Cell Mol Genet. 1986 Mar;12(2):175–183. doi: 10.1007/BF01560664. [DOI] [PubMed] [Google Scholar]
  22. Moore K. A., Deisseroth A. B., Reading C. L., Williams D. E., Belmont J. W. Stromal support enhances cell-free retroviral vector transduction of human bone marrow long-term culture-initiating cells. Blood. 1992 Mar 15;79(6):1393–1399. [PubMed] [Google Scholar]
  23. Morgan R. A., Cornetta K., Anderson W. F. Applications of the polymerase chain reaction in retroviral-mediated gene transfer and the analysis of gene-marked human TIL cells. Hum Gene Ther. 1990 Summer;1(2):135–149. doi: 10.1089/hum.1990.1.2-135. [DOI] [PubMed] [Google Scholar]
  24. Nolta J. A., Kohn D. B. Comparison of the effects of growth factors on retroviral vector-mediated gene transfer and the proliferative status of human hematopoietic progenitor cells. Hum Gene Ther. 1990 Fall;1(3):257–268. doi: 10.1089/hum.1990.1.3-257. [DOI] [PubMed] [Google Scholar]
  25. Nolta J. A., Smogorzewska E. M., Kohn D. B. Analysis of optimal conditions for retroviral-mediated transduction of primitive human hematopoietic cells. Blood. 1995 Jul 1;86(1):101–110. [PubMed] [Google Scholar]
  26. Nolta J. A., Yu X. J., Bahner I., Kohn D. B. Retroviral-mediated transfer of the human glucocerebrosidase gene into cultured Gaucher bone marrow. J Clin Invest. 1992 Aug;90(2):342–348. doi: 10.1172/JCI115868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ojwang J. O., Hampel A., Looney D. J., Wong-Staal F., Rappaport J. Inhibition of human immunodeficiency virus type 1 expression by a hairpin ribozyme. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10802–10806. doi: 10.1073/pnas.89.22.10802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pearson L., Garcia J., Wu F., Modesti N., Nelson J., Gaynor R. A transdominant tat mutant that inhibits tat-induced gene expression from the human immunodeficiency virus long terminal repeat. Proc Natl Acad Sci U S A. 1990 Jul;87(13):5079–5083. doi: 10.1073/pnas.87.13.5079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pomerantz R. J., Trono D. Genetic therapies for HIV infections: promise for the future. AIDS. 1995 Sep;9(9):985–993. doi: 10.1097/00002030-199509000-00002. [DOI] [PubMed] [Google Scholar]
  30. Rhodes A., James W. Inhibition of human immunodeficiency virus replication in cell culture by endogenously synthesized antisense RNA. J Gen Virol. 1990 Sep;71(Pt 9):1965–1974. doi: 10.1099/0022-1317-71-9-1965. [DOI] [PubMed] [Google Scholar]
  31. Sarver N., Cantin E. M., Chang P. S., Zaia J. A., Ladne P. A., Stephens D. A., Rossi J. J. Ribozymes as potential anti-HIV-1 therapeutic agents. Science. 1990 Mar 9;247(4947):1222–1225. doi: 10.1126/science.2107573. [DOI] [PubMed] [Google Scholar]
  32. Schwartz G. N., Kessler S. W., Rothwell S. W., Burrell L. M., Reid T. J., Meltzer M. S., Wright D. G. Inhibitory effects of HIV-1-infected stromal cell layers on the production of myeloid progenitor cells in human long-term bone marrow cultures. Exp Hematol. 1994 Dec;22(13):1288–1296. [PubMed] [Google Scholar]
  33. Su Y., Popik W., Pitha P. M. Inhibition of human immunodeficiency virus type 1 replication by a Tat-activated, transduced interferon gene: targeted expression to human immunodeficiency virus type 1-infected cells. J Virol. 1995 Jan;69(1):110–121. doi: 10.1128/jvi.69.1.110-121.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sullenger B. A., Gallardo H. F., Ungers G. E., Gilboa E. Overexpression of TAR sequences renders cells resistant to human immunodeficiency virus replication. Cell. 1990 Nov 2;63(3):601–608. doi: 10.1016/0092-8674(90)90455-n. [DOI] [PubMed] [Google Scholar]
  35. Szilvassy S. J., Hoffman R. Enriched hematopoietic stem cells: basic biology and clinical utility. Biol Blood Marrow Transplant. 1995 Nov;1(1):3–17. [PubMed] [Google Scholar]
  36. Trono D., Feinberg M. B., Baltimore D. HIV-1 Gag mutants can dominantly interfere with the replication of the wild-type virus. Cell. 1989 Oct 6;59(1):113–120. doi: 10.1016/0092-8674(89)90874-x. [DOI] [PubMed] [Google Scholar]
  37. Vieillard V., Lauret E., Rousseau V., De Maeyer E. Blocking of retroviral infection at a step prior to reverse transcription in cells transformed to constitutively express interferon beta. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2689–2693. doi: 10.1073/pnas.91.7.2689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wells S., Malik P., Pensiero M., Kohn D. B., Nolta J. A. The presence of an autologous marrow stromal cell layer increases glucocerebrosidase gene transduction of long-term culture initiating cells (LTCICs) from the bone marrow of a patient with Gaucher disease. Gene Ther. 1995 Oct;2(8):512–520. [PubMed] [Google Scholar]
  39. Woffendin C., Yang Z. Y., Udaykumar, Xu L., Yang N. S., Sheehy M. J., Nabel G. J. Nonviral and viral delivery of a human immunodeficiency virus protective gene into primary human T cells. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11581–11585. doi: 10.1073/pnas.91.24.11581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yu M., Leavitt M. C., Maruyama M., Yamada O., Young D., Ho A. D., Wong-Staal F. Intracellular immunization of human fetal cord blood stem/progenitor cells with a ribozyme against human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1995 Jan 31;92(3):699–703. doi: 10.1073/pnas.92.3.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zhou C., Bahner I. C., Larson G. P., Zaia J. A., Rossi J. J., Kohn E. B. Inhibition of HIV-1 in human T-lymphocytes by retrovirally transduced anti-tat and rev hammerhead ribozymes. Gene. 1994 Nov 4;149(1):33–39. doi: 10.1016/0378-1119(94)90409-x. [DOI] [PubMed] [Google Scholar]
  42. Zhou C., Bahner I., Rossi J. J., Kohn D. B. Expression of hammerhead ribozymes by retroviral vectors to inhibit HIV-1 replication: comparison of RNA levels and viral inhibition. Antisense Nucleic Acid Drug Dev. 1996 Spring;6(1):17–24. doi: 10.1089/oli.1.1996.6.17. [DOI] [PubMed] [Google Scholar]