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Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy 03/2010 M. Obermeier
Tropism testing from proviral DNA
Analysis of a subgroup from the Berlin Maraviroc cohort
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
HIV-1 Tropism
Macrophage CD4+ CCR5+
Primary T cell CD4+ CCR5+/CXCR4+
T-cell line CD4+ CXCR4+
R5 Virus StrainsM-tropic virusTransmission
R5/X4 Virus StrainsDual-tropic virus
X4 Virus StrainsT-tropic virusEmerge late
non-syncitia-inducing strains (NSI)
syncitia-inducing strains (SI)
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
geno2pheno[coreceptor]
Specificity = 1 - FPR
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Comparison of performance of genotypic tropism testing form viral RNA and proviral DNA
127 viremic patients
VL range 0.5*101 -1.6*106 cop./ml
47 samples were detected CXCR4- tropic using viral RNA
56 samples were detected CXCR4- tropic virus using proviral DNA
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Distribution of FPR in viral and proviral samples
Viral Proviral
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Distribution of CCR5-tropic and CXCR4-tropic detection in viral and proviral samples
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Bland-Altmann analysis of viral and proviral FPR
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Differences in FPR compared to viral load
1 2 3 4 5 6 7
-150
-100
-50
0
50
100
Viral
load
in log cop./ml
ΔFPR
gen
o2ph
eno[
core
cept
or]
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Comparison of Trofile® with geno2pheno[coreceptor] analysis of viral and proviral DNA of 101 samples
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
trofile viral proviral
Trofile R5 g2p X4/X4Trofile R5 g2p X4/R5Trofile R5 g2p R5/X4Trofile R5 vir or pro g2p X4all R5Trofile X4 g2p R5/R5Trofile X4 g2p R5/X4Trofile X4 g2p X4/R5Trofile R5 vir or pro g2p R5all X4
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
The Berlin Maraviroc cohort
157 patients
Reasons for treatment change:
Virological failure
Adverse effects
Tropism Testing
141 (89%) had a genotypic tropism test
95 (60%) had only a genotypic tropism test
53 (32%) had only a genotypic tropism test from poviral DNA
70 (45%) had a Trofile® test (59 Standard Trofile® and 11 Trofile®-ES)
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Results (proviral)
FPR cut-off: 20%
49 patients CCR5
4 patients CXCR4
Baseline:
70% viral load <50 cop./ml
30% low level Virämie (<500 cop./ml)
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Maraviroc cohort FPR
02
46
810
1214
1618
2022
2426
2830
3234
3638
4042
4446
4850
5254
5658
6062
6466
6870
7274
7678
8082
8486
8890
9294
9698100
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
FPR -g2p[coreceptor] false positive rate
Perc
ent C
XCR
4-tro
pic
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Results (proviral)
Week 12 (N= 49)
86% viral load <50 cop./ml
5 patients with virologic failure
1 patient with treatment change due to adverse effects
Week 24 (N=40)
80% viral load <50
4 patients with virologic failure (VL <1000 cop/ml)
1 patient with 60 cop/ml, without treatment change at week 48 below detection limit)
2 patients with treatment change due to adverse effects
Week 48 (N=19)
89% viral load <50 cop./ml
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Results (proviral)
0 12 24 480
10
20
30
40
50
60 >50 cop/ml<50 cop/ml change due to adverse-ef-fects<50 cop/mlNumber of patients
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Patient 153
38 years old, male
HIV diagnosis 1986, Risk MSM
Begin of first ART: 1998
ART-experience: 3TC, ABC, d4T, TDF, FTC, NVP, EFV, SQV, Lopinavir, Ritonavir (fulldose and booster), Darunavir
Last cART: TDF/FTC + NVP
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Patient 153
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Mrz 05
Jun 0
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HIV-load [cop/ml]CD4 cell count [/µl]
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Patient 153
No tropism testing from viral RNA possible
Tropism testing from proviral DNA: CCR5-tropic (FPR 31%)
Genotypic Resistance: No drug resistance associated mutations
Switch to MVC + RAL
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Patient 153
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Mrz 05
Jun 0
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Mrz 09
Jun 0
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HIV-load [cop/ml]CD4 cell count [/µl]
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Patient 153
Integrase-gene: N155H
CXCR4-Tropism detected from viral RNA (FPR: 0.1%)
But:
Patient 74: MVC +RAL
Proviral: CCR5 (25% FPR)
At Week 48 <50 cop/ml
MVC + RAL
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Treatment change in the cohort (proviral subgroup)
Number of drugs in cART
18 patients (34%) received less drugs than before
50 patients (51%) received the same amount of drugs
8 patients (15%) received more drugs
Maximal 8, Minimal 2, Median 3
45 % of the patients received Raltegravir (28% as new drug)
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
What about the CXCR4-tropic proviral results
1 patient with ABC,3TC, DRV/r, RAL + add-on MVC (FPR: 9%) had a viral load of 160 cop./ml at the time of MVC start and 250 cop./ml at week 12
3 other patients (FPR 1%, 16% and 20%) received additional DRV/r or Raltegravir and had a viral load <50 cop/ml at week 12
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Conclusions
Genotypic tropism testing using proviral DNA shows a higher rate of CXCR4 detection as compared to genotypic tropism testing using viral RNA. In combination with a high FPR-cutoff of 20% reasonable safety for the patients can be realized.
Patients from the Berlin Maraviroc cohort that were switched to Maraviroc based on proviral tropism testing showed a high rate of treatment success at weeks 12 and 24.
Due to low patient number harboring a virus with a FPR below 20%, it remains unclear at the moment if lower FPR-cutoff settings than 20% are also feasible.
Presented at the 8th European HIV Drug Resistance Workshop, March 17-19 2010, Sorrento, Italy
Acknowledgement
Physicians in Berlin: Andreas Carganico, Bernhard Bieniek, Christiane Cordes, Stephan Dupke, Klaus Fischer, M Freiwald, Jörg Gölz, Heribert Hillenbrand, Bettina Hintsche, Gerd Klausen, Siegfried Köppe, Peter Kreckel, Elke Lauenroth-Mai, Christoph Mayr, Arend Moll, Stefan Neifer, Daniel Prziwara9, M Rausch, Frank Schlote, Christoph Schuler, Wolfgang Schmidt, Dorothea Schleehauf, Dietmar Schranz, Thomas Wünsche, Axel Baumgarten
Laboratory: Thomas Berg, Stefan Neifer, Andreas Wienbreyer
Pfizer Germany for supporting the physicians in data collection