Detection of hyper-conserved regions in hepatitis B virus X gene potentially useful for gene therapy
González, Carolina (Hospital Universitari Vall d'Hebron)
Tabernero, David (Hospital Universitari Vall d'Hebron)
Cortese, Maria Francesca (Hospital Universitari Vall d'Hebron)
Gregori i Font, Josep (Roche Diagnostics SL)
Casillas, Rosario (Hospital Universitari Vall d'Hebron)
Riveiro Barciela, Mar (Universitat Autònoma de Barcelona. Departament de Medicina)
Godoy, Cristina (Hospital Universitari Vall d'Hebron)
Sopena, Sara (Hospital Universitari Vall d'Hebron)
Rando-Segura, Ariadna (Hospital Universitari Vall d'Hebron)
Yll, Marçal (Hospital Universitari Vall d'Hebron)
Lopez Martinez, Rosa (Hospital Universitari Vall d'Hebron)
Quer, Josep 1963- (Hospital Universitari Vall d'Hebron. Institut de Recerca)
Esteban, Rafael (Esteban Mur) (Universitat Autònoma de Barcelona. Departament de Medicina)
Buti, Maria (Universitat Autònoma de Barcelona. Departament de Medicina)
Rodríguez Frías, Francisco (Universitat Autònoma de Barcelona. Departament de Genètica i de Microbiologia)

Data:	2018
Resum:	To detect hyper-conserved regions in the hepatitis B virus (HBV) X gene (HBX) 5' region that could be candidates for gene therapy. The study included 27 chronic hepatitis B treatment-naive patients in various clinical stages (from chronic infection to cirrhosis and hepatocellular carcinoma, both HBeAg-negative and HBeAg-positive), and infected with HBV genotypes A-F and H. In a serum sample from each patient with viremia > 3. 5 log IU/mL, the HBX 5' end region [nucleotide (nt) 1255-1611] was PCR-amplified and submitted to next-generation sequencing (NGS). We assessed genotype variants by phylogenetic analysis, and evaluated conservation of this region by calculating the information content of each nucleotide position in a multiple alignment of all unique sequences (haplotypes) obtained by NGS. Conservation at the HBx protein amino acid (aa) level was also analyzed. NGS yielded 1333069 sequences from the 27 samples, with a median of 4578 sequences/sample (2487-9279, IQR 2817). In 14/27 patients (51. 8%), phylogenetic analysis of viral nucleotide haplotypes showed a complex mixture of genotypic variants. Analysis of the information content in the haplotype multiple alignments detected 2 hyper-conserved nucleotide regions, one in the HBX upstream non-coding region (nt 1255-1286) and the other in the 5' end coding region (nt 1519-1603). This last region coded for a conserved amino acid region (aa 63-76) that partially overlaps a Kunitz-like domain. Two hyper-conserved regions detected in the HBX 5' end may be of value for targeted gene therapy, regardless of the patients' clinical stage or HBV genotype.
Ajuts:	Instituto de Salud Carlos III PI15/00856
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Hepatitis B virus ; Hepatitis B X gene ; Hepatitis B X protein ; Gene therapy ; Next-generation sequencing ; HBV conserved regions ; Small interference RNA
Publicat a:	World Journal of Gastroenterology, Vol. 24 (may 2018) , p. 2095-2107, ISSN 2219-2840

DOI: 10.3748/wjg.v24.i19.2095
PMID: 29785078

14 p, 1.3 MB

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