Development of a CRISPR/Cas9 system against ruminant animal brucellosis

被引:5
作者
Karponi, Garyfalia [1 ]
Kritas, Spyridon K. [1 ]
Papadopoulou, Gina [2 ]
Akrioti, Elissavet-Kalliopi [3 ]
Papanikolaou, Eleni [4 ,5 ]
Petridou, Evanthia [1 ]
机构
[1] Aristotle Univ Thessaloniki, Fac Hlth Sci, Sch Vet Med, Dept Microbiol & Infect Dis, Thessaloniki 54124, Greece
[2] Acad Athens, Cellular Immunol Lab, Basic Res, Biomed Res Fdn, Athens 11527, Greece
[3] Hellenic Pasteur Inst, Lab Cellular & Mol Neurobiol Stem Cells, Athens 11521, Greece
[4] Univ Athens, Sch Med, Lab Biol, Athens 11527, Greece
[5] Miltenyi Biotec, Dept Res & Dev, D-51429 Bergisch Gladbach, Germany
关键词
Brucellosis; Gene therapy; Viral vectors; Ruminant animals; Macrophages; GENE-THERAPY; LENTIVIRAL VECTOR; MELITENSIS; GENOME; IMMUNODEFICIENCY; INFECTION; VACCINE; SHEEP; SUIS; RNA;
D O I
10.1186/s12917-019-2179-z
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
Background Brucellosis, caused by several Brucella species, such as the bacterium Brucella melitensis, is considered one of the most severe zoonotic diseases worldwide. Not only does it affect ruminant animal populations, leading to a substantial financial burden for stockbreeders, but also poses severe public health issues. For almost four decades in southern Europe and elsewhere, eradication of the disease has been based on ambiguously effective programs, rendering massive sanitation of livestock urgent and indispensable. Gene therapy, which has been proved effective in the clinic, could possibly constitute an alternative option towards a permanent cure for brucellosis, by aiding in the deletion or inactivation of genes associated with the replication of Brucella within the host cells. Results We infected ovine macrophages with B.melitensis, to simulate the host cell/microorganism interaction in vitro, and transduced the infected cells with CRISPR/Cas9 lentiviral vectors that target Brucella's RNA polymerase subunit A (RpolA) or virulence-associated gene virB10 at a multiplicity of infection of 60. We demonstrate a significant decrease in the bacterial load per cell when infected cells are transduced with the RpolA vector and that the number of internalized brucellae per cell remains unaffected when macrophages are transduced with a conventional lentiviral vector expressing the green fluorescence protein, thus underlining the bactericidal effect of our CRISPR/Cas9 system. Conclusions Pending in vivo verification of our findings, overall, these results may prove critical not only for the treatment of human brucellosis, but for other infectious diseases in general.
引用
收藏
页数:10
相关论文
共 42 条
  • [1] The pathology of brucellosis reflects the outcome of the battle between the host genome and the Brucella genome
    Adams, LG
    [J]. VETERINARY MICROBIOLOGY, 2002, 90 (1-4) : 553 - 561
  • [2] Lentiviral Hematopoietic Stem Cell Gene Therapy in Patients with Wiskott-Aldrich Syndrome
    Aiuti, Alessandro
    Biasco, Luca
    Scaramuzza, Samantha
    Ferrua, Francesca
    Cicalese, Maria Pia
    Baricordi, Cristina
    Dionisio, Francesca
    Calabria, Andrea
    Giannelli, Stefania
    Castiello, Maria Carmina
    Bosticardo, Marita
    Evangelio, Costanza
    Assanelli, Andrea
    Casiraghi, Miriam
    Di Nunzio, Sara
    Callegaro, Luciano
    Benati, Claudia
    Rizzardi, Paolo
    Pellin, Danilo
    Di Serio, Clelia
    Schmidt, Manfred
    Von Kalle, Christof
    Gardner, Jason
    Mehta, Nalini
    Neduva, Victor
    Dow, David J.
    Galy, Anne
    Miniero, Roberto
    Finocchi, Andrea
    Metin, Ayse
    Banerjee, Pinaki P.
    Orange, Jordan S.
    Galimberti, Stefania
    Valsecchi, Maria Grazia
    Biffi, Alessandra
    Montini, Eugenio
    Villa, Anna
    Ciceri, Fabio
    Roncarolo, Maria Grazia
    Naldini, Luigi
    [J]. SCIENCE, 2013, 341 (6148) : 865 - U71
  • [3] Gene Therapy for Immunodeficiency Due to Adenosine Deaminase Deficiency.
    Aiuti, Alessandro
    Cattaneo, Federica
    Galimberti, Stefania
    Benninghoff, Ulrike
    Cassani, Barbara
    Callegaro, Luciano
    Scaramuzza, Samantha
    Andolfi, Grazia
    Mirolo, Massimiliano
    Brigida, Immacolata
    Tabucchi, Antonella
    Carlucci, Filippo
    Eibl, Martha
    Aker, Memet
    Slavin, Shimon
    Al-Mousa, Hamoud
    Al Ghonaium, Abdulaziz
    Ferster, Alina
    Duppenthaler, Andrea
    Notarangelo, Luigi
    Wintergerst, Uwe
    Buckley, Rebecca H.
    Bregni, Marco
    Marktel, Sarah
    Valsecchi, Maria Grazia
    Rossi, Paolo
    Ciceri, Fabio
    Miniero, Roberto
    Bordignon, Claudio
    Roncarolo, Maria-Grazia
    [J]. NEW ENGLAND JOURNAL OF MEDICINE, 2009, 360 (05) : 447 - 458
  • [4] Bakhrebah MA, 2018, EUR REV MED PHARMACO, V22, P3448, DOI 10.26355/eurrev_201806_15169
  • [5] Control of small ruminant brucellosis by use of Brucella melitensis Rev.1 vaccine:: laboratory aspects and field observations
    Banai, M
    [J]. VETERINARY MICROBIOLOGY, 2002, 90 (1-4) : 497 - 519
  • [6] Lentiviral Hematopoietic Stem Cell Gene Therapy Benefits Metachromatic Leukodystrophy
    Biffi, Alessandra
    Montini, Eugenio
    Lorioli, Laura
    Cesani, Martina
    Fumagalli, Francesca
    Plati, Tiziana
    Baldoli, Cristina
    Martino, Sabata
    Calabria, Andrea
    Canale, Sabrina
    Benedicenti, Fabrizio
    Vallanti, Giuliana
    Biasco, Luca
    Leo, Simone
    Kabbara, Nabil
    Zanetti, Gianluigi
    Rizzo, William B.
    Mehta, Nalini A. L.
    Cicalese, Maria Pia
    Casiraghi, Miriam
    Boelens, Jaap J.
    Del Carro, Ubaldo
    Dow, David J.
    Schmidt, Manfred
    Assanelli, Andrea
    Neduva, Victor
    Di Serio, Clelia
    Stupka, Elia
    Gardner, Jason
    von Kalle, Christof
    Bordignon, Claudio
    Ciceri, Fabio
    Rovelli, Attilio
    Roncarolo, Maria Grazia
    Aiuti, Alessandro
    Sessa, Maria
    Naldini, Luigi
    [J]. SCIENCE, 2013, 341 (6148) : 864 - U58
  • [7] A review of the use of B-melitensis Rev 1 vaccine in adult sheep and goats
    Blasco, JM
    [J]. PREVENTIVE VETERINARY MEDICINE, 1997, 31 (3-4) : 275 - 283
  • [8] Hematopoietic Stem Cell Gene Therapy with a Lentiviral Vector in X-Linked Adrenoleukodystrophy
    Cartier, Nathalie
    Hacein-Bey-Abina, Salima
    Bartholomae, Cynthia C.
    Veres, Gabor
    Schmidt, Manfred
    Kutschera, Ina
    Vidaud, Michel
    Abel, Ulrich
    Dal-Cortivo, Liliane
    Caccavelli, Laure
    Mahlaoui, Nizar
    Kiermer, Veronique
    Mittelstaedt, Denice
    Bellesme, Celine
    Lahlou, Najiba
    Lefrere, Francois
    Blanche, Stephane
    Audit, Muriel
    Payen, Emmanuel
    Leboulch, Philippe
    l'Homme, Bruno
    Bougneres, Pierre
    Von Kalle, Christof
    Fischer, Alain
    Cavazzana-Calvo, Marina
    Aubourg, Patrick
    [J]. SCIENCE, 2009, 326 (5954) : 818 - 823
  • [9] Gene therapy of human severe combined immunodeficiency (SCID)-X1 disease
    Cavazzana-Calvo, M
    Hacein-Bey, S
    Basile, CD
    Gross, F
    Yvon, E
    Nusbaum, P
    Selz, F
    Hue, C
    Certain, S
    Casanova, JL
    Bousso, P
    Le Deist, F
    Fischer, A
    [J]. SCIENCE, 2000, 288 (5466) : 669 - 672
  • [10] Gene silencing by CRISPR interference in mycobacteria
    Choudhary, Eira
    Thakur, Preeti
    Pareek, Madhu
    Agarwal, Nisheeth
    [J]. NATURE COMMUNICATIONS, 2015, 6