Title Page
Contents
국문초록 11
ABSTRACT 13
I. Introduction 15
1. Osteoporosis 15
2. Bisphosphonate and treatment failure 16
3. Osteo-immune system and single cell RNA sequencing 17
4. Hypothesis 18
II. Objectives 19
III. Materials and Methods 19
1. Study design and inclusion/exclusion criteria 19
2. Hospital data collection 21
3. Peripheral blood mononuclear cell isolation and cryopreservation 23
4. Single-cell library preparation and sequencing 24
5. scRNA seq data processing 24
6. Unsupervised clustering and cluster annotation 25
7. Gene network implication 25
8. Statistical analysis and soft ware/data processing 26
IV. Results 27
1. Incidence of bisphosphonate treatment failure 27
2. Radiological and morphological data 28
3. scRNA seq for included patients 33
4. TCR+ macrophage subset enrichment in osteoporosis[이미지참조] 36
5. Gene expression analysis in monocyte subtypes 37
6. Heterogeneity of T cell subtypes and NK cells 39
7. Heterogeneity of B cell subtypes 44
8. Heterogeneous gene expression of B cell subtypes 45
V. Discussion 47
VI. Conclusion 52
References 53
Appendix 63
Table 1. Demographic data in this study. 28
Table 2. Radiological analysis in this study. 30
Table 3. Morphological analysis in this study. 31
Table 4. Univariate and multivariate regression analyses 32
Figure 1. Radiological and morphological parameters in this study. 23
Figure 2. Incidence rate of bisphosphonate treatment failure in postmenopausal patients with osteoporotic vertebral fractures. 27
Figure 3. Receiver-operating characteristic (ROC) curve analysis for initial bone mineral density (BMD) of the spine and initial FRAX hip. 33
Figure 4. Single-cell transcriptome profiling of peripheral blood mononuclear cells (PBMCs) from normal and osteoporosis patients including bisphosphonate (BP)-... 35
Figure 5. Monocytes cluster profiling and composition of subtypes. 37
Figure 6. Expression of featured genes in monocyte subtypes. 39
Figure 7. T·NK cells cluster profile and dynamic composition of subtype. 40
Figure 8. Analysis of NK cluster and differentially expressed genes in bisphosphonate (BP)-failure. 42
Figure 9. Expression of featured genes in T·NK subtypes. 44
Figure 10. Profiling B cells cluster and composition of subtypes. 45
Figure 11. Expression of featured genes in B cell subtypes. 46