arv induces long-acting effects? neurological symptoms azt +/- 3tc blanche et al lancet 1999...
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ARV induces long-acting effects?
• Neurological symptoms AZT +/- 3TC Blanche et al Lancet 1999
• Persistent hyperlactatemia AZT +/- 3TCNoguera et al Ped Inf Dis J 2003
• Altered hematopoïesis AZT +/- 3TC Lechenadec et al AIDS 2003
• Altered myocardal contractility AZT +/- 3TC Lipshultz et al J Am Coll Cardiology 2011
Neutrophils count in perinatally AZT exposed children until 8 years
AZT exposed
ECS AIDS 2005;19:1071
AZT unexposed
AZT-induced long-lasting impairment
of hematopoieisis
Is there hematopoietic Is there hematopoietic stem cell CD34+ genotoxic stem cell CD34+ genotoxic
alteration ?alteration ?I Schmutz Andre, E Six, M Cavazzana-Calvo, S Kaltenbach F Mavillio, Paris and Modena
Lechenadec AIDS 2003, ECS AIDS 2004, Pacheco J Inf Dis 2006…etc
Validation by qRT-PCR
Cord blood cells CD34+ cells
Affymetrix Gene chip
analysisBioinformatic
analysis
Transcriptional profiling analysis
of CD34+ hematopoietic
stem cells
HIV neg AZT- exposedand control newborn
Gene expression in CD34+ cord blood cells of AZT exposed newborn
Maternités : A. Béclère, L. Mourrier, Bichat, Lariboisière, Cochin, Necker.
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Cord blood cell immunophenotyping: hematopoietic stem cells (HSC)
and multipotent progenitors(MPP )
Lin-CD38-CD34+CD90-Lin-CD38-CD34+CD90+
CD34+
213 genes up-regulated in ARV-treated cells
90 genes down-regulated in ARV-treated cells
ND vs ARV pvalue 0.05 Fold Change 1.5
Gene expression profilesControl vs AZT-exposed CD34+
Genes functionsIngenuity® functional clustering analysis
Cellular Growth and Proliferation
Cell Death
Cell Cycle
DNA Replication, Recombination and Repair
ALOX12 ALOX12 CCL5AXL CASP1 AXL CCNE1BAMBI CASP3 CCNE1 CD180C7orf68 CCL2 CCNE2 IL1BCASP1 CCL5 CDK1 ITGB3CASP3 CCR1 DDX17 MDM2CCL2 CDK1 EGF ORC6CCL5 CTTN GDF15 PDE5ACCNE1 EGF H19 PDGFACCNE2 GDF15 HMGA2 TFPI2CCR1 GIMAP5 ID1 VPRBPCD36 HMGA2 ID2 WRNCDK1 ICAM1 IL1B COL4A1 ID1 ITGB3 DDX17 IL1B KLHL2
EGF ITGB3 KNTC1 GBP1 ITGB5 MDM2 GDF15 LGALS8 MDM4 HMGA2 MDM2 ORC6 ICAM1 MDM4 PDK3 ID1 NFIL3 PRKCA ID2 PDGFA PTAFR IL1B PF4 RPS6KA2 ITGA2B PPBP VPRBP ITGB3 PRKCA WRN ITGB5 PTAFR KDM4C MDM2 MDM4 PDE5A PDGFA PF4 PLD1 PRKCA PTAFR PTCH1 PTGS1 PTPRJ TFPI2 VPRBP WRN
The effect involves particularly regulator genes of chromosomal instability
– centrosome amplification (Cyclin E1 and E2)
– checkpoint defect (Rae1)
– dysregulation of cell-cycle proteins (Mdm2) – microtubule attachment and chromosome
segregation (TUBB, MARK4)
These major cell cycle regulators
are involved in chromosome segregation
Stem Cells and DNA Damage:Persist or Perish?
A. Lane Cell 2010;142, 360–362
Increased aneuploidy in cord blood lymphocytes from AZT-exposed newborn
Proportional increase of micronuclei
Median (%): 6.6 vs 18.7 p<0.001
Summary
• These findings are reminiscent of cellular and animal AZT exposure experiments
• Aneuploidy appears as the result of DNA damage induced by AZT integration
• They confirm significant in vivo genotoxicity in cord blood cells after exposure to AZT in utero
• Unknown long-term clinical consequences
Signification clinique ?
• Hématopoïèse à long terme ?
• Risque de cancer hematopoïétique ?
• Autre cancer ?
AZT in utero exposure and risk of cancer
To date , no global increased risk
• 9127 AZT or AZT+3TC exposed children
• Median age : 5.4 y
• >53 000 patient- year
• 10 cases observed vs 9.6 expected
French Cohort . Benhammou AIDS 2008
6 nouveaux cas de cancers repérés depuis l’analyse de 2007
Nouveau croisement avec registre SFCE prévu
Suites….
• Données de Primeva
• Comparaison de différentes combinaisons d’ARV avec ces outils moléculaires– Observationnelles– Randomisées ?