표제지

목차

국문 초록 7

서론 9

다발성 경화증의 발병 기전 9

장 내 마이크로바이옴과 신경 병증 10

다발성 경화증과 EAE 유도 마우스 모델의 장 내 미생물 연구 12

Veillonella의 기능 14

재료 및 방법 16

1. Veillonella strains 배양 16

2. Whole genome Pacbio sequencing 분석 17

3. EAE 유도 동물 모델 실험 및 샘플 수집 18

4. 마우스 모델 장관 마이크로바이옴 분석법 최적화 19

5. Metagenome 추출 및 metagenome shotgun sequencing 분석 20

6. 미생물 총 수 정량을 위한 Quantitative real-time qPCR 21

7. 통계분석 22

ⅰ. Decontamination 22

ⅱ. Indicator analysis 22

ⅲ. MaAsLin2 analysis 23

ⅳ. Intra-, inter-variation analysis 23

ⅴ. Network analysis 23

ⅵ. Sankey diagram 24

ⅶ. Multiscale-PHATE & MELD analysis 24

ⅷ. Statistical comparison and correlation analysis 24

결과 25

마우스 모델 장관 마이크로바이옴 분석법 최적화 25

Veillonella MHL0042 섭취를 통한 마우스의 EAE 중증도 완화 28

EAE 유도에 따른 장관 미생물의 dysbiosis 33

Veillonella MHL0042에 의한 맹장 및 대장 마이크로바이옴 변화 42

고찰 52

참고문헌 57

영문 초록 62

Table 1. Mouse GI tract bacteria mock community genome list. 26

Table 2. EAE clinical scoring chart based on the behavior of EAE-induced mice. 31

Figure 1. Evaluation of a mouse GI tract microbiome analysis pipeline using a mock community. (a) Evaluation of bacteria classification accuracy using... 27

Figure 2. Design for the EAE-induced mouse model experiment and EAE clinical score. (a) A total of 20 C57BL/L6 female mice were used in the experiment. The PBS and EAE groups were fed PBS for the duration... 30

Figure 3. Pan-genome analysis using Pacbio sequencing data. (a) Veillonella pan-genome analyzed by the Anvi'o pipeline. The core and unique genes of Veillonella MHL0042, MHL0057, MHL0067, and ATCC17746... 32

Figure 4. Verify Veillonella MHL0042 engraftment in mouse GI tract. (a) The relative abundance of Veillonella MHL0042 in mouse fecal metagenome after... 37

Figure 5. Comparison of microbiome between mouse model GI tract. (a) The diversity of microbiota and bacterial amounts among mouse model GI tract... 38

Figure 6. Dysbiosis of the mouse GI tract microbiota by EAE. (a) Shifts of indicator species for each GI tract were caused by EAE induction. The indicator's relative... 39

Figure 7. Significant difference of KO 3rd category pathway among the EAE status. (a) Volcano plot analysis shows the different KO 3rd category pathway among the...[이미지참조] 40

Figure 8. Difference of microbiota composition and diversity according to the EAE status in PSI, DSI, colon, and fecal. (a) Microbiota composition of species-level... 41

Figure 9. Network analysis of the EV groups microbiota community in the cecum. Positive correlations are marked by red edges and negative correlations by blue... 45

Figure 10. Satellite species, positively correlated with keystone, in microbial network of the cecum. Boxplots show the relative abundance among the EAE... 46

Figure 11. Analysis of the gene families that encoded the flagellum in the cecum. (a) The Sankey diagram shows which flagellar components are encoded by gene... 47

Figure 12. Network analysis of the EV groups microbiota community in the colon. Positive correlations are marked by red edges and negative correlations by blue... 48

Figure 13. Keystones translocate from the cecum to the colon. (a) Multiscale potential of heat diffusion for affinity-based transition embedding (MS-PHATE)... 49

Figure 14. Satellite species, positively correlated with keystone, in microbial network of the colon. Boxplots show the relative abundance among the EAE status... 50

Figure 15. Analysis of the gene families that encoded the flagellum in the colon. (a) The Sankey diagram shows which flagellar components are encoded by gene... 51

Figure 16. Graphical summary 56