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Metagenomic Analysis of Skin Microbiome and Circulating Bacterial Extracellular Vesicles in Atopic Dermatitis

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Authors

김민혜

Advisor
조상헌
Major
의과대학 협동과정임상약리학전공
Issue Date
2015-08
Publisher
서울대학교 대학원
Keywords
Atopic dermatitisStaphylococcus aureusLactic acid bacteriaMetagenomics
Description
학위논문 (박사)-- 서울대학교 대학원 : 협동과정 임상약리학전공, 2015. 8. 조상헌.
Abstract
Introduction: Staphylococcus aureus is the most common bacterium colonizing the skin of atopic dermatitis (AD) patients. Up to 90% of those with AD are colonized by S. aureus, which is much higher than the rate in healthy subjects. In addition to the contribution of local bacteria, systemic bacterial structure affects pathogenesis of AD. No previous study has considered the systemic bacterial contribution. Until recently, bacteria could only be identified by culture-based assays, while identify only 1% of the total bacteria and which takes days. Here, we performed metagenomic analysis to determine the proportion of colonizing or pathogenic bacteria in the local or systemic environment of AD patients compared to healthy controls.

Methods: Twenty-seven patients with AD and 6 healthy control subjects were enrolled. Skin washing fluids were obtained from moistened gauze positioned on skin lesions of AD patients and the cubital fossa of normal controls. Blood and urine samples were also obtained. Genomic DNA was extracted from all samples. The 16s ribosomal RNA gene was amplified using the universal primer and sequenced using a next generation sequencer. Sequenced data were analyzed.

Results: S. aureus was dominant in the skin from AD patients and hardly detected in skin from control subjects. Staphylococcus comprised 66.2% and 0.2% of the total bacterial DNA of AD patients and normal controls, respectively. Alcaligenaceae family and Sediminibacterium were the most frequent bacteria in the skin of the controls. After treatment, the proportion of Staphylococcus decreased from 38.8% to 5.7%. The proportion of Alicyclobacillus, Propionibacterium, and Streptococcus was increased from 2.2% to 6.0%, from 2.2% to 4.6%, and from 0.5% to 6.5%, respectively, in the post-treatment group. The bacterial composition of urine and serum were almost identical, and both differed appreciably from skin. The proportion of Lactococcus, Leuconostoc, and Lactobacillus were significantly increased in the control group, and Alicyclobacillus, Propionibacterium, and Streptophyta(o) were increased in AD patients.

Conclusions: Metagenomics of bacterial DNA in the skin washing fluid of AD patients confirmed that S. aureus is a main skin colonizer of AD compared to normal controls and that the bacterial composition changes dramatically after treatment. S. aureus may be closely related to the pathogenesis and exacerbation of AD. Lactic acid bacteria, which are beneficial constituents of the human microbiome, were dominant in the urine of healthy controls compared to AD patients. This indicates that lactic acid bacteria might have protective roles against AD.
Language
English
URI
https://hdl.handle.net/10371/121789
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College of Medicine/School of Medicine (의과대학/대학원)Program in Clinical Pharmacology (협동과정-임상약리학전공)Theses (Ph.D. / Sc.D._협동과정-임상약리학전공)
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