Mixed HCV Genotype Infection and Response to anti-HCV treatment in HIV/HCV Co-

Infected PatientsLucy Porrino, Sabrina Bagaglio, Giulia Morsica, Giulia Gallotta, Hamid

Hasson, Laura Galli,Vega Rusconi, Adriano Lazzarin, Caterina Uberti-Foppa

Infectious Diseases Dept, San Raffaele Scientific Institute, Milan, Italy

WEAB0102

Background• The frequency of mixed infection in HCV monoinfection is

about 2-13% (Schroter et al. 2003; Qian et al. 2000)

• There is a single study on viral infections with different genotypes in HIV/HCV coinfected patients and has not been evaluated the performance of the genotype during the anti-viral therapy (Soriano et al. 2005)

• Previous reports from Japan identified a mutational pattern within the core protein (70Q-91M) of HCV-1b related to the responsiveness to standard treatment for HCV

• This finding possibly mediated by an interaction with interferon signalling cellular protein STAT-1

To determine the HCV genotype pattern and core domain in HIV/HCV coinfected patients during anti-HCV treatment

AIM

Study Patients• A pilot clinical trial (Kamon2) was conducted to compare

LPV/r monotherapy and LPV/r-based HAART during anti-HCV therapy

• A virological sub-study was performed on patients HIV/HCV coinfected enrolled in Kamon2

• Patients were analyzed according to response to anti-HCV treatment

HCV-RNA negativity at 24-weeks of treatment and maintainment up to 72 weeks

Sustained Virological Response = (SVR)

≤2 Log decrease of HCV-RNA at 12-weeks

No Response (NR) =

HCV-RNA positivity during follow up Relapse (RE) post-treatment =

Methods

• Sequence analysis of HCV-5’ UTR was performed in plasma at different time points: baseline (BL), during 48 weeks treatment and after treatment

• The entire sequence of core was inferred in plasma at BL, during treatment and/or after treatment. All these sequences were compared with the respective prototype (based on HCV genotyping)

Baseline Characteristics of HIV/HCV Coinfected Individuals According to Anti-HCV Treatment

ResponseAll patients

(N=22)SVR

(N=11)NR(NR+RE)

(N=11)P

(SVR vs NR)

Age (years) 42 (40-44) 44 (41-45) 42 (39-43) 0.95

Males/Females

13/9 6/5 7/4 1

ALT (U/I) 79.5 (59-164) 95 (74-150) 59 (45-137) 0.77

AST (U/I) 48 (36-88) 51 (41-84) 39 (29-83) 0.80

CD4+cells count

(cell/mmc)

592 (414-794)

596 (421-739)

514 (407-921)

0.70

CD8+cells count

(cell/mmc)

925 (693-1167)

926 (837-1116)

935 (595-1258)

0.70

HCV-RNA (Log IU/ml)

6.05 (4.83-6.34)

5.37(4055-6.03)

6.34 (6.11-6.42)

0.05

HCV Genotype (1-4 vs 2-3)

13/9 2/9 0/11 0.0002

HCV Genotype Pattern in NR during Follow-Up

BL W4 W12 W24 W36 W48 W60 W72

NR 1 1a NA 3a

NR 2 4a 3a 4a 4a 4c/d

NR 3 1b NA 1b 3a NA NA 1 1b

NR 4 1b 1b 3a 2a 1b

RE 1 1b 3a NA neg neg neg neg 1b

6 (55%) maintained the same HCV genotype during follow up

5 (45%) showed an alternance of HCV genotype in plasma

lost in follow up

lost in follow up

lost in follow up

Core sequence analysis at BL

• We obtained the core sequences of 10 SVR and 8 NR

• Different distribution of amino acids (aa) mutations in different genotype (G)

HCV Genotype (n) Median number of aa mutations (IQR)

G1 (7) 1 (0-4.5)G2 (2) 4 (3-5)G3 (6) 6 (6-12)G4 (3) 4 (4-4.50)

G1-G4 vs G2-G3 p= 0.0276

G1 vs G3 p= 0.018

Core sequence analysis during follow up in 8 NR patients

• 1pt showed the same sequence at each time point • 7 pts showed at least one aa substitution during

follow up• This aa mutation were recurrent in specific position

aapositio

n20 36 70 71 75 110

N°pts 6/8 3/8 2/8 3/8 6/8 5/8

...in particular

• 6/8 showed at least aa substitution in 1-20* and 69-75 **region

• 5/8 showed at least aa substitution in 110-115* region

110 120 130 140 ....|....|....|....|....|....|....|....|...

.| RGSRPSWGPTDPRRRSRNLGKVIDTLTCGFADLMGYIPLVGAPLG

10 20 30 40 50 ....|....|....|....|....|....|....|....|....|....|MSTNPKPQRKTKRNTNRRPQDVKFPGGGQIVGGVYLLPRRGPRLGVRATR

60 70 80 90 100....|....|....|....|....|....|....|....|....|....|KTSERSQPRGRRQPIPKARRPEGRTWAQPGYPWPLYGNEGCGWAGWLLSP

*epitope regions**peptide-specific CD4+ T-cell

• 5/8 pts showed 70Q and/or 91M» 70Q was undetectable at BL in 3pts

and appeared during treatment» in 1pt this mutation was detectable at

BL and disapperead during treatment» 1pt had 91M at BL and maintained

during treatment

Characteristic mutational pattern in 2 RE at BL...

NR vs RE p= 0.0056

...and disappeared during follow up

Conclusions - 5’ UTR analysis

• The dynamic pattern of HCV genotypes in half of NR suggests a mixed HCV-infection

• The anti-HCV treatment may induce a change balance of dominant strain

• Work in progress: ultradeep sequencing (pyrosequencing) will be performed to verify viral population in plasma

Conclusions - Core domain analysis

• At BL difficult to treat genotypes are more similar to the respective prototype than “favourable”genotypes

• On treatment the most prominent variability of specific residues in specific regions of the core domain could be consequent to the pressure exerted by interferon and/or ribavirin

Thanks to• Infectious Diseases Dept, San Raffaele Scientific Institute,

MilanGiulia MorsicaSabrina BagaglioMarco MerliAlice DaddaHamid HassonGiulia GallottaLaura GalliVega RusconiCaterina Uberti-FoppaAdriano Lazzarin

• National Institute of Infectious Diseases, Spallanzani, Roma

• Infectious Diseases Section, Hospital SS Annunziata, Firenze

• Institute of Infectious Diseases, University of Bari, Bari