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Drug Resistance Mutations in HIV-1
isolates from HIV-1-infected patients in Morocco
S. MRANI, MD, PhD
Virology Laboratory, Mohammed V Military Teaching Hospital-Rabat
Faculty of Medicine and Pharmacy-Mohammed V University -Rabat
Background
•
In 2015 HIV-infection remained a significant public health issue
•
ONUSIDA : 35 millions of personnes with HIV
UNAIDS/WHO. The Gap Report 2014
2
Backgroud
•
This region of Africa is home to only 10% of the world population while it
represents 70% of people living with HIV in the world
•
Demographic ties between Morocco and sub-Saharan Africa:
-active cooperation with African countries
-Moroccans residing or staying long in those countries
3
In our context
 10633 VIH cases
 5243 (60%) AIDS
Cases of HIV/AIDS Marocco
from 1986 to 2015
Moroccan Ministery of Health. HIV National Report 2015.
4
CONTEXTE
Moroccan Ministery of Health. HIV National Report 2015.
5
CONTEXTE
HIV Treatment as Prevention
(TasP)
What about HIV Drug Resistance?
6
Antiretroviral drugs
PR Inhibitors (PIs) (9)
Nucleoside/nucleotide RT Inhibitors (NRTIs) (8)
Non-nucleoside RT Inhibitors (NNRTIs) (4)
Fusion Inhibitors (2)
Integrase Inhibitors (1)
CCR5 Inhibitors
 Current therapies imperfect because:
Regimen complexities
Toxicities
Drug resistance
Background
8
Background
9
How drug resistance arises
How drug resistance arises. Richman, DD. Scientific American , July 1998
Evolution of Viral Mutations
 Mutations arise because HIV-1 RT makes
spontaneous errors (1 in 104)
 HIV-1 genome is 10 000 (104) bases long, therefore
1 error each time the genome is replicated
 Production of virus = 109 to 1010 virions per day 
quasispecies
 Every possible mutation present in quasispecies
before ARV therapy
How do you measure drug resistance?
Phenotyping:
Direct assay: Measures the ability of the virus to grow in
various concentrations of antiretroviral drugs.
Genotyping:
Indirect assay: Detects drug resistance mutations that
are present in the relevant virus genes.
Phenotyping
13
Genotyping
14
OBJECTIVE
Determine ARV resistance mutations in HIV-1-infected
antiretroviral treatment naïve patients
15
Patients
CHAPITRE
et Méthods
II
CHAPITRE
Patients
 HIV-1-infected and antiretroviral treatment naïve patients were
included in this study
 They followed-up at the Mohammed V Military Teaching Hospital in
Rabat between 2008 and 2011
The study was approved by the bioethical committee of the faculty
of medicine and pharmacy of Rabat.
12/04/2017
16
Patients
et Méthods
CHAPITRE
I
Méthodes
The RNA extraction was carried out using the High Pure Viral RNA Kit
(Roche Diagnostics Systems).
The viral RNA was used in reverse transcription polymerase chain reaction
(RTPCR) followed by a nested PCRof reverse transcriptase (RT) and
protease genes
The obtained fragments were sequenced on both strands using the CEQ
DTCS Quick Start kit on an automated sequencer Beckman Coulter
GenomeLab GeXP DNA Analyzer System.
12/04/2017
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Patients
CHAPITRE
et Méthods
II
CHAPITRE
Méthodes
The derived nucleotide sequences of the RT and protease regions were
aligned by the Clustal W 1.74 alignment program
Phylogenetic trees with bootscanning methods were inferred using the
neighbor-joining method.
 GenBank accession numbers for the sequences reported
in this study are HQ393323 to HQ393404 and HQ393240 to HQ393322 for
RT and protease sequences, respectively.
The mutations involved in antiretroviral resistance transmission were
checked according to the international list of surveillance drug-resistance
mutations (SDRMs) updated in 2011.
12/04/2017
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RESULTS I
CHAPITRE
12/04/2017
Charactéristics
Nomber (91)
Age
Moyenne (years)
34 (19-57)
%
Sex
Men
Women
71
20
78
22
Transmission
Heterosexuel
unknown
81
10
89
11
Clinique Stage (CDC)
A
B
C
55
13
23
60
15
25
CD4 Count (cells/ml)
Médiane (percentile 25–75)
116 [80-385]
viral load (copies/ml)
Médiane (percentile 25–75)
149.050 [49450-336000]
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RESULTS
Phylogenic analysis
RESULTS
Code du patient
Mutation Protéase
Mutation TI
Sous-type
2611/08
-
184V
B
6325/05
-
215S
B
7720/07
82A
103N.184V
C
9844/09
-
103N.184V.215I
B
Discussion (1)
 Our patients diagnosed with advanced HIV infection in the 40% in stage
B or C to 116 CD4 cells / l
similar data reported by other studies in Morocco
The majority subtype B: 74%
Non-B strains accounted for 26% (15% CRF02_AG, 6% A1, 2% C, 1% F1,
1% and 1% CRF09 CRF25-cpx)
Several studies have reported that diversity Mediterranean.
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Discussion (2)
Contries
Time of study
MDR Prevalence
Morocco (casablanca)
2004-2007
5%
Italy
2004-2008
15.7%
Espagne
1997-2008
10.6%
Grèce
2002-2003
9%
France(chroniquement infectés)
2007-2012
9%
France (primo-infection )
2007-2012
12.2 %
Burkina Faso
2003-2009
8.3%
Congo
2004-2010
4.2%
Uganda
2000-2010
< 3%
Our Study
2007-2014
5.06%
23
Limitations of Drug Resistance Testing
1) Relationship between drug resistance and clinical failure is
complex
 Non-adherence
 Sub-optimal regimens
 Pharmacokinetics
 Host factors
 Many drug resistant variants are less fit
2) Complex quasispecies
3) Cross-resistance
4) Potentially miss minor populations
Conclusion
 Resistance-conferring mutations occur continuously, in
absence and presence of drug therapy.
 Occur more rapidly when viral load is high: more
replication, more mutations.
 Resistance testing gives information on which drugs no
longer potent in regimen.
 Drug resistance knowledge is important for interpreting
genotypic resistance tests, designing surveillance studies,
and drug development.
 Understanding resistance is important for optimal patient
management