2nd International Workshop on Adverse Drug Reactions and Lipodystrophy in HIV 13-15 September 2000, Toronto, Canada

SUBCUTANEOUS ADIPOSE TISSUE MITOCHONDRIAL DNA ANALYSIS FROM INDIVIDUALS WITH HAART-ASSOCIATED LIPODYSTROPHY

Antiviral Therapy 2000; 5(Suppl. 5):6 (abstract no. O7)

C Shikuma, N Hu, C Milne and B Shiramizu
Center of Clinical Research Excellence, Research Centers in Minority Institutions, University of Hawaii at Manoa, Honolulu, Hawaii, USA

BACKGROUND: Recent studies have speculated that NRTI-induced mitochondrial toxicity may be the initial result from HAART leading to fat redistribution. In addition to physical changes, the implication of fat tissue mitochondrial toxicity impacts overall metabolic imbalance. Therefore a detailed analysis of tissue-specific mitochondrial DNA (mtDNA) was undertaken to determine the characteristics of mtDNA from fat biopsies.

OBJECTIVES: The objective was to analyse mtDNA from HIV-infected individuals on HAART with/without self-reported lipodystrophy; and from controls and to detect the presence/absence of mtDNA and use restriction-digestion analysis.

DESIGN: Subcutaneous fat by core-needle biopsies were obtained from individuals following informed consent from neck, abdomen and thigh. mtDNA was extracted and long-distance PCR was used to amplify mtDNA and semiquantitate the mtDNA. Appropriate positive and negative controls were used. Restriction digests were performed using EcoRV and SacI.

RESULTS: To date, 36 biopsies have been analysed from 12 individuals (seven HIV-infected and five non- HIV-infected individuals). PCR detection for mtDNA was able to demonstrate the presence or absence of mtDNA as well as semiquantify the amount of the mtDNA present. Two of seven HIV-infected individuals had clinical history of fat redistribution. The mtDNA analysis of these individuals showed absence/decreased amounts of mtDNA in 4/6 specimens (66.7% of specimens). The five HIV-infected individuals with no history of lipodystrophic changes showed absence or decreased mtDNA in 3/15 specimens (20.0%). From the control specimens, mtDNA was absent or decreased in 4/15 specimens (26.7%). To date, six specimens from two non-HIV-infected individuals have been analysed with restriction digestion with no deletion or insertion mutations noted.

CONCLUSIONS: Preliminary data suggest absence/ decreased amounts of mtDNA in subcutaneous adipose tissue of HIV-infected individuals with lipodystrophy, which is consistent with possible mitochondrial toxicity. Further analysis of specific mutations from the tissue-specific areas are needed to determine if deletion/insertion mutations are present in the mtDNA.

Supported by: Pacific Biomedical Research Center RCMI/HARC Grant RR/A103061 and CCRE Grant RR 99005.

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