Mutational analysis of the mitochondrial tRNA genes and flanking regions in umbilical cord tissue from uninfected infants receiving AZT-based therapies for prophylaxis of HIV-1.
A sensitive vertical denaturing gradient gel electrophoresis (DGGE) method, using 13 unipolar psoralen-clamped PCR primer pairs, was developed for detecting sequence variants in the 22 tRNA genes and flanking regions (together spanning approximately 21%) of the human mitochondrial genome. A study was conducted to determine (i) if mitochondrial DNA (mtDNA) polymorphisms and/or mutations were detectable in healthy newborns and (ii) if prepartum 3'-azido-2',3'-dideoxythymidine (AZT) based HIV-1 prophylaxis was associated with significant increases in mtDNA mutations and changes in the degree of heteroplasmy of sequence variants in uninfected infants born to HIV-1-infected mothers. DGGE analysis of umbilical cord tissue (where vascular endothelium and smooth muscle cells are the major source of mtDNA) showed that mtDNA sequence variants were significantly elevated by threefold in AZT-treated infants compared with unexposed controls (P < 0.001), with 24 changes observed in 19/52 (37%) treated newborns (averaging 0.46 changes/subject) versus only eight changes found in 7/55 (13%) unexposed newborns (averaging 0.15 changes/subject). Six distinct sequence variants occurring in unexposed controls were predominately synonymous and homoplasmic, representing previously reported polymorphisms. Uninfected infants exposed to a combination of AZT and 2',3'-dideoxy-3'-thiacytidine and "maternal HIV-1" had a significant shift in the spectrum of mutations (P = 0.04) driven by increases in nonsynonymous heteroplasmic sequence variants at polymorphic sites (10 distinct variants) and novel sites (four distinct variants). While the weight of evidence suggests that prepartum AZT-based prophylaxis produces mtDNA mutations, additional research is needed to determine the degree to which fetal responses to maternal HIV-1 infection, in the absence of antiretroviral treatment, contribute to prenatal mtDNA mutagenesis.
Torres
Salina M
SM
Lovelace Respiratory Research Institute, Albuquerque, New Mexico, USA.
Walker
Dale M
DM
McCash
Consuelo L
CL
Carter
Meghan M
MM
Ming
Jessica
J
Cordova
Edmund M
EM
Pons
Rachel M
RM
Cook
Dennis L
DL
Seilkop
Steven K
SK
Copeland
William C
WC
Walker
Vernon E
VE
D001483:Base Sequence; D016022:Case-Control Studies; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics*; D004359:Drug Therapy, Combination; D015658:HIV Infections / Q000517:prevention & control*; D015497:HIV-1; D006801:Humans; D007223:Infant; D019259:Lamivudine / Q000008:administration & dosage / Q000627:therapeutic use*; D009154:Mutation; D011110:Polymorphism, Genetic; D012343:RNA, Transfer / Q000235:genetics*; D018894:Reverse Transcriptase Inhibitors / Q000008:administration & dosage / Q000627:therapeutic use*; D014470:Umbilical Cord / Q000378:metabolism*; D015215:Zidovudine / Q000008:administration & dosage / Q000627:therapeutic use*
D017931:DNA Primers; D004272:DNA, Mitochondrial; D018894:Reverse Transcriptase Inhibitors; D019259:Lamivudine; D015215:Zidovudine; D012343:RNA, Transfer