Combination therapy of 5-FU and α-galactosylceramide enhances the anti-tumor effect in CT26 cancer model
- 주제(키워드) Mesenchymal stem cell , cytosine deaminase , 5-Fluorocytosine , α-galactosylceramide , immunotherapy
- 주제(DDC) 570
- 발행기관 아주대학교
- 지도교수 Haeyoung Suh-Kim
- 발행년도 2023
- 학위수여년월 2023. 8
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000032837
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Cancer immunotherapy has emerged as a promising approach for treating various malignant tumors. In this study, we performed a screening for the most effective PAMP molecule when combined with mesenchymal stem cells expressing cytosine deaminase (MSC/CD) and 5-fluorocytosine (5-FC) in a colon cancer model, resulting in the identification of α-galactosylceramide (α-GalCer). In addition, we determined the optimal timing and frequency of administration for maximized therapeutic effects when used in combination therapy. Our findings demonstrate that the combination of MSC/CD, 5-FC, and α-GalCer induces enhanced antitumor activity compared to individual treatments. This enhancement is primarily due to the increased immune response incited by the administration of α-GalCer. In particular, intratumoral injection of α-GalCer increases natural killer T (NKT) cell infiltration and activation into the tumor microenvironment. These findings lead to the infiltration and activation of immune cells within the tumor microenvironment, including natural killer (NK) cells, T cells, and antigen presenting cells (APCs) such as dendritic cells (DCs) and M1 macrophages. In addition, proinflammatory and chemokine levels in tumors increase due to the activation of immune cells. IFN-γ functions as a key central cytokine, and tumor cell death is increased through increased Granzyme B with 5-FU. Therefore, the combined administration of MSC/CD, 5-FC, and α-GalCer effectively enhances both innate and adaptive immune responses, thereby amplifying anticancer efficacy. We also observed no hepatotoxicity induced by α-GalCer. The systemic inflammation and increase in anti-inflammatory cytokines were found to be transient reactions, indicating the safety of our combined approach in the context of immunotherapy. Our research underscores the potential therapeutic benefits of integrating MSC/CD, 5-FC, and α-GalCer for colon cancer treatment, and contributes valuable insights to the field of cancer immunotherapy. Future studies should be focused on elucidating the fundamental mechanisms and exploring the possibility of applying these findings to other types of cancer and immune therapeutic strategies.
more목차
A. INTRODUCTION 1
B. MATERIALS AND METHODS 9
1. Cell culture and maintenance 9
2. Animals 9
3. Reagent preparation 10
4. 5-FU sensitivity test 10
5. CT26 cancer model 10
6. Evaluation of the tumor cell killing effect of 5-FU in vivo 10
7. Hematoxylin and eosin (H&E) staining 11
8. Bystander effect test 11
9. Evaluation of anti-cancer effect by combined administration of MSC/CD and various doses of 5-FC in CT26 cancer model 12
10. Evaluation of in vivo therapeutic efficacy of MSC/CD, 5-FC in CT26 cancer model 12
11. Evaluation of anticancer effects by combined administration of MSC/CD, 5-FC, and various PAMP molecules in the CT26 cancer model 12
12. Evaluation of appropriate dosing timing and frequency of α-GalCer for combination with MSC/CD and 5-FC in CT26 cancer Model 13
13. Evaluation of in vivo therapeutic efficacy of MSC/CD, 5-FC, and α-GalCer in CT26 cancer Model 13
14. Evaluation of white blood cell (WBC) changes 14
15. Measurement of in vivo cytokines 14
16. Measurement of in vivo immune cells 15
17. Assessment of hepatoxicity by α-GalCer 15
18. Evaluation of anticancer effect by administration of MSC/CD, 5-FC, and/or α-GalCer in BNL cancer model 16
19. Quantification and statistical analysis 16
C. RESULTS 18
1. Cell death effect of 5-FU in CT26 cells 18
2. MSC/CD with 5-FC showed highly effective cell death in CT26-GFP cells 21
3. MSC/CD with 5-FC demonstrates moderate in vivo anticancer effects in a colon cancer model 24
4. α-GalCer is an immunomodulator that has a synergic effect with MSC/CD with 5-FC 27
5. Combination of MSC/CD with 5-FC and α-GalCer exhibits enhanced anticancer effects in a colon cancer model 29
6. Administration of MSC/CD and α-GalCer affects NKT cells and intratumoral cytokines 35
7. Effects of MSC/CD and/or α-GalCer administration on intratumoral antigen-presenting cells (APCs) and cytokine levels 39
8. Effects of MSC/CD and α-GalCer administration on CD4+ T cell and CD8+ T cell counts and cytotoxicity of these cells 43
9. Effects of MSC/CD and α-GalCer Administration on Immunosuppressive Cells and Cytokine Levels 48
10. The Enhanced Anticancer Efficacy of α-GalCer Alone in the BNL hepatocarcinoma model 52
11. Toxicity evaluation of α-GalCer and combined administration of α-GalCer and MSC/CD 54
D. DISCUSSION 62
E. REFERENCES 67
국문요약 76
