Presentation on theme: "ARV failure and resistance for the paediatrician"— Presentation transcript:
1ARV failure and resistance for the paediatrician Douglas Watson, M.D.University of Maryland11 December 2013
2REVERSE TRANSCRIPTASE INHIBITORS (NRTIs & NNRTIs) Integrase inhibitorsReverse transcriptase inhibitors (nucleoside and non-nucleoside RT inhibitors) prevent the synthesis of a DNA copy of the viral RNA genome. Protease inhibitors act on the virus after it buds through the cell membrane, inhibiting the cutting of long inactive polypeptide chain into shorter functional proteins that the virus needs to infect the next cell. As our bodies do not perform reverse transciption, inhibiting that step should not adversely affect us. But many of the HIV nucleoside reverse transcriptase inhibitors do have an effect on the enzyme that replicates mitochondrial DNA- accounting for the toxicity of the NRTIs. There is one fusion inhibitor in clinical use, but it is extremely expensive to make. More useful integrase inhibitors show promising early results in humans.Fusion InhibitorsPROTEASE INHIBITORSAttachment InhibitorsHIV DRUG TARGETS
4WHO 2013 recommendations for 1st-line ART in children < 3 y of age “ABC should be considered the preferred NRTI whenever possible.”LPV//r should not be given to infants < 14 d of age or in premature infants until after 14 d after their due date
5WHO 2013: Start with LPV/r and switching to NNRTI? NOTE:The quoted study enrolled only children with history of SD NVP exposureThe quoted study used < 50 copies/ml as primary endpoint and < 1,000 c/ml as secondary endpoint- NOT < 400 c/ml as stated above.The switch-to-NVP group had fewer cases of VL > 50 but more cases of VL > 1,000 and more resistance than group that stayed on LPV/r
6WHO 2013 recommendations for 1st line ART in children > 3 y of age
7Treatment failure: progressive steps, different definitions Potency failureAdherence failureVirologic failureGenotypic failure (resistance)There are multiple definitions or types of treatment failure. If the regimen is inadequate or the dosage too low (potency failure), or if there is nonadherence (adherence failure), then virologic failure (a detectable viral load) will develop. Genotypic failure will develop, the time course depending on the genetic barrier to resistance and how high the viral load is- the lower barrier to resistance and higher the viral load, the more rapidly resistance is selected. With higher levels of viral load, eventually the CD4 stops going up and begins to fall (immunologic failure). Even later, the patient experiences renewed symptoms- clinical failure.Immunologic failureClinical failure
8Expected fall in viral load TimeLog fold drop in viral loadFold-drop in viral loadViral load (example)--600,000100,0001 month2100+< 6,000< 1,0003 months31,0006001006 months410,00060<50If viral load at these follow up times is 3-fold or more than these examples, full suppression not likely
9Days = 0 is day HAART started (6 weeks of age) Days = 0 is day HAART started (6 weeks of age). Note initial rapid rise in viral load after infection at birth. Also not 2-phase decay in viral load on HAART- an initial rapid drop (about 1000-fold) in first 2-3 weeks, then slower decline to undetectable by ultrasensitive assay by 140 days of treatment. This girl is now 8 years old and has had an undetectable viral load since about 4 months of age.
10Course of treatment failure Start ZDV/3TC/NVPClinical deteriorationGradual ZDV resistanceNonadherenceNVP resistance3TC resistanceCD4 CountViral loadThe time course of treatment failure. The VL drops very rapidly after starting HAART, then more slowly. At the indicated point, the patient has a period of nonadherence. Very rapidly, NVP resistance (K103N mutation) is selected. What follows from this point will occur whether or not adherence improves. Within 1-3 months, 3TC resistance (M184V) is selected. This increases sensitivity to ZDV, but at this point ZDV is the only active drug- not enough to hold back the virus. The CD4 count has continued to improve, even after development of NVP resistance- we know that ZDV/3TC will make someone with AIDS better for a year or so. With time, however, the CD4 stops rising and starts to fall. One by one, ZDV-selected thymidine analogue mutations evolve. (TAMS: RT mutations at 41, 67, 69, 70, 210, 211, 215 and others- 215 and 41 being the most important.) Only as the viral load gets very high again and the CD4 has fallen does the patient have recurrent symptoms. At this point there is resistance to all 3 drugs with cross-resistance to EFV and complete or partial cross-resistance to all NRTIs.1 year2 years
11Mechanism of reverse transcriptase inhibitors cDNA NRTI = Nucleoside reverse transcriptase inhibitorNNRTI = Non-nucleoside reverse transcriptase inhibitorHIV reverse transcriptasecDNAABCHIV reverse transcriptase copies the viral RNA into DNA. The nucleoside analogues (D4T in this case), are added onto the growing DNA chain. But because they are blocked at one end, the next natural dNTP cannot be added- and the DNA chain is terminated. The NNRTIs work by binding to the RT molecule, blocking its activity.NRTI (such as ABC(P3) is added onto cDNA chain, blocking further reverse transcriptionNNRTI (NVP or EFV)Blocks reverse transcriptase by binding at active site
12The K103N mutation: how it can take over Sensitive K103 HIV wild type (wt)Resistant 103N HIVNVP blocks sensitive HIV, not resistant HIVNevirapinewt always presentNVP blocks the grown of the wild-type virus (with K at position 103). But the extremely rare, randomly produced mutant, with N at position 103, is able to grow. Within 2-4 weeks the mutant resistant virus takes over as the predominant virus in the blood.In presence of NVP, only the resistant virus grows. Soon almost all virus is resistant!
13Note that these patients were failing (by VL monitoring) for only 4-5 months- which is too early to detect failure using CD4 counts or clinical signs- yet they had developed significant resistance- nearly all NNRTI and 3TC resistance, and significant amount of other NRTI mutations. Note that these other mutations were more common in patients with VL > 4 log (> 10,000) copies/ml.Drug resistance mutations in Thai patients failing D4T/3TC/NVP for an average of 4-5 months by viral load monitoring. Sungkanuparph S. CID 2007; 44:447–52
16NNRTI resistance 1st generation (nevirapine, efavirenz, delavirdine) High potency but low genetic barrier to resistanceMost commonly K103N- resistance to all 1st generationNVP also selects for Y181C (especially in newborns) which has mild effect on EFV but associated with increased failuresOther mutation patterns also seen2nd generation (Etravirine)NO EFFECT of K103NResistance increases with other NNRTI mutations- 3 or more yield
18Protease inhibitor resistance Some PIs select for drug-specific mutations (e.g. NFV, ATV)Some PIs can be boosted or unboosted (ATV, fAPV)- low-level resistance may be clinically significant if not boostedBoosted PIs more durableResistance to LPV requires 5-10 mutationsVirologic failure while receiving LPV/r usually due to nonadherenceProlonged virologic failure while on LPV/r eventually will lead to LPV resistance
19Drug resistance testing Commercial methods start with RT PCR of bulk virus in plasmaNot sensitive to minor strains- e.g. genotyping (sequencing) cannot detect strain representing < 10-25% of circulating virusWhen to get resistance testing (resource-rich)Baseline: resistant strains (especially NNRTI resistance) circulating in populationWhenever resistance suspected (e.g. failing and patient appears adherent)Selective pressure- patient taking medication
20HIV drug resistance genotype RT PCR bulk plasma virus to produce cDNASequence pol geneDerive predicted amino acid sequenceIdentify mutations known to confer resistance (e.g. Stanford database, IAS-USA, etc.)Virtual phenotype™ genotype interpretationIdentified set of significant mutations is matched with massive database of genotype-phenotype correlationsReported as predicted fold resistance (used to also give number of matches)Fold-resistance interpreted according to in vitro or clinical measure of activity
26Interpretation of resistance testing Complete treatment and viral load history are essentialWhat is current regimen and is patient taking it?Many resistance mutations cause decreased viral fitness: if there is not selective pressure, wild-type virus rapidly outgrows mutant, but archive of mutant remains
28Johnny B. Goode Newly infected adolescent CD4 = 320, VL = 58,000 Prescribed TDF/FTC/ATV/rVL 1 month later = 5,200VL at 2 months = 4,000Genotype is wild-typeHow do you interpret this situation?How to proceed?