Development of Gene Editing Strategies for CCR5 Gene in Endothelial Cells

Author :  

DOI : 10.29228/genediting.54872
Year-Number: 2021-2
Yayımlanma Tarihi: 2021-12-08 12:25:15.0
Language : English
Konu : Medical Biology
Number of pages: 22-30
Mendeley EndNote Alıntı Yap

Abstract

The CRISPR system has been adapted as a promising gene editing technology that provides robust and efficient DNA alterations. It was demonstrated that it has a high adaptability and great potential to cure acquired immune deficiency syndrome (AIDS) through gene disruptions. Endothelial cells are an essential component of the vascular system, and CRISPR has emerged as a promising candidate for AIDS therapies by removing the human immunodeficiency virus (HIV) reservoir in endothelial cells. CCR5, an essential receptor for HIV infection, is expressed by endothelial cells. We developed CCR5 gRNAs and tested their effectiveness in gene targeting. We established a basic and successful transfection system, as well as on-target CCR5 mutagenesis using the endonuclease assay. These findings imply that endothelial diseases could be targeted using the CCR5 gRNA/CRISPR system, which would be a novel approach. Endothelial diseases caused by genetic mutations or HIV infection could be treated in endothelial cells using the CRISPR/Cas9-based gene therapy platform by targeting CCR5.

Keywords

Abstract

The CRISPR system has been adapted as a promising gene editing technology that provides robust and efficient DNA alterations. It was demonstrated that it has a high adaptability and great potential to cure acquired immune deficiency syndrome (AIDS) through gene disruptions. Endothelial cells are an essential component of the vascular system, and CRISPR has emerged as a promising candidate for AIDS therapies by removing the human immunodeficiency virus (HIV) reservoir in endothelial cells. CCR5, an essential receptor for HIV infection, is expressed by endothelial cells. We developed CCR5 gRNAs and tested their effectiveness in gene targeting. We established a basic and successful transfection system, as well as on-target CCR5 mutagenesis using the endonuclease assay. These findings imply that endothelial diseases could be targeted using the CCR5 gRNA/CRISPR system, which would be a novel approach. Endothelial diseases caused by genetic mutations or HIV infection could be treated in endothelial cells using the CRISPR/Cas9-based gene therapy platform by targeting CCR5.

Keywords

Kaynakça

  • 1. Control CfD. Update on acquired immune deficiencysyndrome (AIDS)--United States. MMWR Morbidity and mortality weekly report. 1982;31(37):507.
  • 2. Bayer R, Levine C, Murray TH. Guidelines forConfidentiality in Research on AIDS. AIDS Res. 1983;1(4):275- 97.
  • 3. Cunningham AL, Donaghy H, Harman AN, Kim M,Turville SG. Manipulation of dendritic cell function by viruses. Curr Opin Microbiol. 2010;13(4):524-9.
  • 4. Klasse PJ. The molecular basis of HIV entry. Cell Microbiol. 2012;14(8):1183-92.
  • 5. Deng H, Liu R, Ellmeier W, Choe S, Unutmaz D,Burkhart M, et al. Identification of a major co-receptor for primary isolates of HIV-1. Nature. 1996;381(6584):661-6.
  • 6. Samson M, Libert F, Doranz BJ, Rucker J, Liesnard C,Farber CM, et al. Resistance to HIV-1 infection in caucasianindividuals bearing mutant alleles of the CCR-5 chemokine receptor gene. Nature. 1996;382(6593):722-5.
  • 7. Liu R, Paxton WA, Choe S, Ceradini D, Martin SR,Horuk R, et al. Homozygous defect in HIV-1 coreceptoraccounts for resistance of some multiply-exposed individuals to HIV-1 infection. Cell. 1996;86(3):367-77.
  • 8. Dean M, Carrington M, Winkler C, Huttley GA, SmithMW, Allikmets R, et al. Genetic restriction of HIV-1 infectionand progression to AIDS by a deletion allele of the CKR5structural gene. Hemophilia Growth and Development Study,Multicenter AIDS Cohort Study, Multicenter Hemophilia CohortStudy, San Francisco City Cohort, ALIVE Study. Science. 1996;273(5283):1856-62.
  • 9. Moore RD, Chaisson RE. Natural history of HIVinfection in the era of combination antiretroviral therapy. AIDS. 1999;13(14):1933-42.
  • 11. Allers K, Hutter G, Hofmann J, Loddenkemper C,Rieger K, Thiel E, et al. Evidence for the cure of HIV infection byCCR5Delta32/Delta32 stem cell transplantation. Blood.12. Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA,Charpentier E. A programmable dual-RNA-guided DNAendonuclease in adaptive bacterial immunity. Science.13. Zhang F, Wen Y, Guo X. CRISPR/Cas9 for genomeediting: progress, implications and challenges. Hum Mol Genet. 2014;23(R1):R40-6.
  • 14. Kim YG, Cha J, Chandrasegaran S. Hybrid restrictionenzymes: zinc finger fusions to Fok I cleavage domain. Proc Natl Acad Sci U S A. 1996;93(3):1156-60.
  • 15. Perez EE, Wang J, Miller JC, Jouvenot Y, Kim KA, LiuO, et al. Establishment of HIV-1 resistance in CD4+ T cells bygenome editing using zinc-finger nucleases. Nat Biotechnol.16. Ding Q, Regan SN, Xia Y, Oostrom LA, Cowan CA,Musunuru K. Enhanced efficiency of human pluripotent stemcell genome editing through replacing TALENs with CRISPRs. Cell Stem Cell. 2013;12(4):393-4.
  • 17. Doudna JA, Charpentier E. Genome editing. The newfrontier of genome engineering with CRISPR-Cas9. Science. 2014;346(6213):1258096.
  • 18. Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE,et al. RNA-guided human genome engineering via Cas9. Science.19. Lieber MR. The mechanism of double-strand DNAbreak repair by the nonhomologous DNA end-joining pathway. Annu Rev Biochem. 2010;79:181-211.
  • 20. Wang W, Ye C, Liu J, Zhang D, Kimata JT, Zhou P.CCR5 gene disruption via lentiviral vectors expressing Cas9 andsingle guided RNA renders cells resistant to HIV-1 infection. PLoS One. 2014;9(12):e115987.
  • 21. Hutter G, Bodor J, Ledger S, Boyd M, Millington M,Tsie M, et al. CCR5 Targeted Cell Therapy for HIV and Prevention of Viral Escape. Viruses. 2015;7(8):4186-203.
  • 22. Wilen CB, Wang J, Tilton JC, Miller JC, Kim KA, RebarEJ, et al. Engineering HIV-resistant human CD4+ T cells withCXCR4-specific zinc-finger nucleases. PLoS Pathog. 2011;7(4):e1002020.
  • 23. Hou P, Chen S, Wang S, Yu X, Chen Y, Jiang M, et al.Genome editing of CXCR4 by CRISPR/cas9 confers cells resistant to HIV-1 infection. Sci Rep. 2015;5:15577.
  • 24. Yu S, Yao Y, Xiao H, Li J, Liu Q, Yang Y, et al.Simultaneous Knockout of CXCR4 and CCR5 Genes in CD4+ TCells via CRISPR/Cas9 Confers Resistance to Both X4- and R5-Tropic Human Immunodeficiency Virus Type 1 Infection. Hum Gene Ther. 2018;29(1):51-67.
  • 25. Mandal PK, Ferreira LM, Collins R, Meissner TB,Boutwell CL, Friesen M, et al. Efficient ablation of genes inhuman hematopoietic stem and effector cells using CRISPR/Cas9. Cell Stem Cell. 2014;15(5):643-52.
  • 26. Palella FJ, Jr., Baker RK, Moorman AC, Chmiel JS,Wood KC, Brooks JT, et al. Mortality in the highly activeantiretroviral therapy era: changing causes of death and diseasein the HIV outpatient study. J Acquir Immune Defic Syndr.27. Crum NF, Riffenburgh RH, Wegner S, Agan BK, TaskerSA, Spooner KM, et al. Comparisons of causes of death andmortality rates among HIV-infected persons: analysis of the pre-,early, and late HAART (highly active antiretroviral therapy) eras. J Acquir Immune Defic Syndr. 2006;41(2):194-200.
  • 29. Sackoff JE, Hanna DB, Pfeiffer MR, Torian LV. Causesof death among persons with AIDS in the era of highly activeantiretroviral therapy: New York City. Ann Intern Med.30. Mateen FJ, Shinohara RT, Carone M, Miller EN,McArthur JC, Jacobson LP, et al. Neurologic disorders incidencein HIV+ vs HIV- men: Multicenter AIDS Cohort Study, 1996- 2011. Neurology. 2012;79(18):1873-80.
  • 31. French AL, Gawel SH, Hershow R, Benning L, HessolNA, Levine AM, et al. Trends in mortality and causes of deathamong women with HIV in the United States: a 10-year study. J Acquir Immune Defic Syndr. 2009;51(4):399-406.
  • 32. Mocroft A, Reiss P, Gasiorowski J, Ledergerber B,Kowalska J, Chiesi A, et al. Serious fatal and nonfatal non-AIDS-defining illnesses in Europe. J Acquir Immune Defic Syndr.33. Currier JS, Lundgren JD, Carr A, Klein D, Sabin CA, SaxPE, et al. Epidemiological evidence for cardiovascular disease inHIV-infected patients and relationship to highly active antiretroviral therapy. Circulation. 2008;118(2):e29-35.
  • 34. Triant VA, Lee H, Hadigan C, Grinspoon SK. Increasedacute myocardial infarction rates and cardiovascular risk factorsamong patients with human immunodeficiency virus disease. J Clin Endocrinol Metab. 2007;92(7):2506-12.
  • MA, Valdivielso P, et al. Peripheral arterial disease in HIVpatients older than 50 years of age. AIDS Res Hum Retroviruses.36. Periard D, Cavassini M, Taffe P, Chevalley M, Senn L,Chapuis-Taillard C, et al. High prevalence of peripheral arterialdisease in HIV-infected persons. Clin Infect Dis. 2008;46(5):761- 7.
  • 37. Cotrim C, Almeida AG, Carrageta M. Exercise-inducedintra-ventricular gradients as a frequent potential cause ofmyocardial ischemia in cardiac syndrome X patients. Cardiovasc Ultrasound. 2008;6:3.
  • 38. Gibellini D, Borderi M, Clo A, Morini S, Miserocchi A,Bon I, et al. HIV-related mechanisms in atherosclerosis andcardiovascular diseases. J Cardiovasc Med (Hagerstown).39. Hsue PY, Hunt PW, Schnell A, Kalapus SC, Hoh R,Ganz P, et al. Role of viral replication, antiretroviral therapy, andimmunodeficiency in HIV-associated atherosclerosis. AIDS.40. Gokce N, Keaney JF, Jr., Hunter LM, Watkins MT,Nedeljkovic ZS, Menzoian JO, et al. Predictive value ofnoninvasively determined endothelial dysfunction for long-termcardiovascular events in patients with peripheral vascular disease. J Am Coll Cardiol. 2003;41(10):1769-75.
  • 41. Suwaidi JA, Hamasaki S, Higano ST, Nishimura RA,Holmes DR, Jr., Lerman A. Long-term follow-up of patients withmild coronary artery disease and endothelial dysfunction. Circulation. 2000;101(9):948-54.
  • 42. Funderburg NT, Mayne E, Sieg SF, Asaad R, Jiang W,Kalinowska M, et al. Increased tissue factor expression oncirculating monocytes in chronic HIV infection: relationship toin vivo coagulation and immune activation. Blood.43. Hansen L, Parker I, Sutliff RL, Platt MO, Gleason RL, Jr.Endothelial dysfunction, arterial stiffening, and intima-mediathickening in large arteries from HIV-1 transgenic mice. Ann Biomed Eng. 2013;41(4):682-93.
  • 44. Solages A, Vita JA, Thornton DJ, Murray J, Heeren T,Craven DE, et al. Endothelial function in HIV-infected persons. Clin Infect Dis. 2006;42(9):1325-32.
  • 45. Oh SK, Cruikshank WW, Raina J, Blanchard GC, AdlerWH, Walker J, et al. Identification of HIV-1 envelopeglycoprotein in the serum of AIDS and ARC patients. J Acquir Immune Defic Syndr (1988). 1992;5(3):251-6.
  • 46. Schneider J, Kaaden O, Copeland TD, Oroszlan S,Hunsmann G. Shedding and interspecies type sero-reactivity ofthe envelope glycopolypeptide gp120 of the humanimmunodeficiency virus. J Gen Virol. 1986;67 ( Pt 11):2533-8.
  • 47. Fiala M, Popik W, Qiao JH, Lossinsky AS, Alce T, TranK, et al. HIV-1 induces cardiomyopathyby cardiomyocyteinvasion and gp120, Tat, and cytokine apoptotic signaling. Cardiovasc Toxicol. 2004;4(2):97-107.
  • 48. Huang MB, Khan M, Garcia-Barrio M, Powell M, BondVC. Apoptotic effects in primary human umbilical veinendothelial cell cultures caused by exposure to virion- associatedand cell membrane-associated HIV-1 gp120. J Acquir Immune Defic Syndr. 2001;27(3):213-21.
  • 49. Ullrich CK, Groopman JE, Ganju RK. HIV-1 gp120-and gp160-induced apoptosis in cultured endothelial cells is mediated by caspases. Blood. 2000;96(4):1438-42.
  • 50. Kanmogne GD, Kennedy RC, Grammas P. HIV-1 gp120proteins and gp160 peptides are toxic to brain endothelial cellsand neurons: possible pathway for HIV entry into the brain andHIV-associated dementia. J Neuropathol Exp Neurol. 2002;61(11):992- 1000.
  • 51. Khan NA, Di Cello F, Stins M, Kim KS. Gp120-mediated cytotoxicity of human brain microvascular endothelialcells is dependent on p38 mitogen-activated protein kinase activation. J Neurovirol. 2007;13(3):242-51.
  • 52. Wang H, Yang H, Shivalila CS, Dawlaty MM, ChengAW, Zhang F, et al. One-step generation of mice carryingmutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell. 2013;153(4):910-8.
  • 53. Daigneault BW, Rajput S, Smith GW, Ross PJ.Embryonic POU5F1 is Required for Expanded Bovine Blastocyst Formation. Sci Rep. 2018;8(1):7753.
  • 54. Tripathi JN, Ntui VO, Ron M, Muiruri SK, Britt A,Tripathi L. CRISPR/Cas9 editing of endogenous banana streakvirus in the B genome of Musa spp. overcomes a major challenge in banana breeding. Commun Biol. 2019;2:46.
  • et al. A foundation for universal T-cell based immunotherapy: Tcells engineered to express a CD19-specific chimeric-antigen-receptor and eliminate expression of endogenous TCR. Blood.59. Voit RA, McMahon MA, Sawyer SL, Porteus MH.Generation of an HIV resistant T-cell line by targeted "stacking" of restriction factors. Mol Ther. 2013;21(4):786-95.
  • JAMA Cardiol. 2017;2(8):914-9.
  • 61. Kocherova I, Bryja A, Mozdziak P, Angelova Volponi A,Dyszkiewicz-Konwinska M, Piotrowska-Kempisty H, et al.Human Umbilical Vein Endothelial Cells (HUVECs) Co-Culturewith Osteogenic Cells: From Molecular Communication toEngineering Prevascularised Bone Grafts. J Clin Med. 2019;8(10).62. Daniels BP, Cruz-Orengo L, Pasieka TJ, Couraud PO,Romero IA, Weksler B, et al. Immortalized human cerebralmicrovascular endothelial cells maintain the properties ofprimary cells in an in vitro model of immune migration acrossthe blood brain barrier. J Neurosci Methods. 2013;212(1):173-9.
  • 63. Stone NL, England TJ, O'Sullivan SE. A Novel TranswellBlood Brain Barrier Model Using Primary Human Cells. Front Cell Neurosci. 2019;13:230.
  • 64. Thomas P, Smart TG. HEK293 cell line: a vehicle for theexpression of recombinant proteins. J Pharmacol Toxicol Methods. 2005;51(3):187-200.
  • 65. Boussif O, Lezoualc'h F, Zanta MA, Mergny MD,Scherman D, Demeneix B, et al. A versatile vector for gene andoligonucleotide transfer into cells in culture and in vivo:polyethylenimine. Proc Natl Acad Sci U S A. 1995;92(16):7297-
MAKALE İSTATİSTİKLERİ

                                                                                                                                                                                                        

                                                                                                                                                                                                        

                                                                                                                                                                                                        
                                                                                                                                                                                                        
                                                                                                                                                                                                        

                                                                                                                                                                                                        

                                                                                                                                                                                                        



                                                                                                                                                                                                        

                                                                                                                                                                                                        
 
                                                                                                                                                                                                        
                      
                                                                                                                                                                                                        

                                                                                                                                                                                                        

                                                                                                                                                                                                        
LINKS

                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
  • 
                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
    

                                                                                                                                                                                                        
     Last issue
    
                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
    • 
                                                                                                                                                                                                        
  • 
                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
         Previous issues
    
                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
    • 
                                                                                                                                                                                                        
  • 
                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
    Article Statistics
    
                                                                                                                                                                                                        
    
                                                                                                                                                                                                        
    • 

                                                                                                                                                                                                        
                                                                                                                                                                                                        




                                                                                                                                                                                                        


                                                                                                                                                                                                        

                                                                                                                                                                                                        



                                                                                                                                                                                                        

                                                                                                                                                                                                        

                                                                                                                                                                                                        
Share