AGGF1中和抗体联合抗CTLA-4抗体抑制小鼠色素瘤生长的分子机制研究
Study on Molecular Mechanism of AGGF1 Neutralizing Antibody Combined with Anti CTLA-4 Antibody in Suppressing the Growth of Mouse Melanoma
DOI: 10.12677/acm.2024.1482396, PDF,    国家自然科学基金支持
作者: 李 夏：桂林医学院智能医学与生物技术学院，广西 桂林；桂林医学院广西脑与认知神经科学重点实验室，广西 桂林；牧苏婉, 杨惠元, 韦妙灵, 韦柳青, 李 勇*：桂林医学院智能医学与生物技术学院，广西 桂林
关键词: 黑色素瘤；移植性模型；AGGF1；CTLA-4；中和抗体；联合治疗；Melanoma； Transplantable Model； AGGF1； CTLA-4； Neutralizing Antibodies； Combination Therapy
摘要: 目的：探究抗AGGF1中和抗体(RDD-Ab)，RDD-Ab + 抗CTLA-4抗体联合治疗对小鼠黑色素瘤的治疗作用。方法：合成AGGF1的GTFQRDDAPASVHSE肽并制备多克隆中和抗体(RDD-Ab)。为了评估RDD-Ab对血管生成活性和黑色素瘤生长的影响，我们实施了小管生成实验、迁移实验、细胞增殖实验、黑色素瘤细胞皮下移植性模型实验和免疫组化等实验。结果：实验结果显示，制备的RDD-Ab可以识别细胞中天然的AGGF1蛋白和过表达的AGGF1蛋白。RDD-Ab可显著抑制血管内皮细胞小管形成、迁移和增殖。与IgG对照组相比，RDD-Ab治疗显著减缓黑色素瘤生长，RDD-Ab + CTLA-4抗体联合治疗时肿瘤生长速度最慢。免疫组化实验也表明，RDD-Ab显著减少瘤内微血管生成与肿瘤细胞增殖，同时，联合治疗可显著增加瘤内CD4+和CD8+淋巴细胞的浸润。结论：RDD-Ab可在体外抑制血管内皮细胞的血管新生功能，具有高效的黑色素瘤生长抑制作用，RDD-Ab + CTLA-4联合治疗黑色素瘤效果更佳(显著增加瘤内CD4+和CD8+淋巴细胞的浸润)，这为未来黑色素瘤的临床干预提供了一种新的潜在治疗方案。
Abstract: Objective: To investigate the therapeutic effect of neutralizing antibody (against AGGF1, RDD-Ab), RDD-Ab + CTLA-4 therapy on mouse melanoma. Methods: Synthesize GTFQRDAPASVHSE peptide of AGGF1 and prepare polyclonal neutralizing antibody (RDD-Ab). In order to evaluate the effects of RDD-Ab on angiogenesis and melanoma growth, we conducted tube formation, migration, cell proliferation, subcutaneous melanoma cell transplantation model and immunohistochemistry (IHC). Results: RDD-Ab can recognize both natural AGGF1 protein and overexpressed AGGF1 protein in cells. The experimental results showed that RDD-Ab significantly inhibits the formation, migration, and proliferation of endothelial cell tubules. Compared with the IgG control group, RDD-Ab significantly slowed down the growth of melanoma. IHC experiments showed that RDD-Ab significantly suppressed tumor angiogenesis and proliferation. The combination therapy of RDD-Ab + CTLA-4 antibody has the slowest tumor growth rate, and the combination therapy increases the infiltration of CD4+ and CD8+ lymphocytes robustly in solid tumors. Conclusion: RDD-Ab can inhibit the angiogenesis function of endothelial cells in vitro and has a highly effective inhibitory effect on melanoma growth has an efficient inhibitory effect on melanoma growth, and the combined treatment with CTLA-4 antibody is more effective. This provides a new potential treatment option for clinical intervention of melanoma in the future.
文章引用：李夏, 牧苏婉, 杨惠元, 韦妙灵, 韦柳青, 李勇. AGGF1中和抗体联合抗CTLA-4抗体抑制小鼠色素瘤生长的分子机制研究[J]. 临床医学进展, 2024, 14(8): 1601-1609. https://doi.org/10.12677/acm.2024.1482396

参考文献

[1] Parhi, R., Kaishap, P.P. and Jena, G.K. (2023) Recent Advances in Nanomaterial-Based Drug Delivery Systems for Melanoma Therapy. ADMET and DMPK, 12, 107-150. [Google Scholar] [CrossRef] [PubMed]
[2] Arnold, M., Singh, D., Laversanne, M., Vignat, J., Vaccarella, S., Meheus, F., et al. (2022) Global Burden of Cutaneous Melanoma in 2020 and Projections to 2040. JAMA Dermatology, 158, 495-503. [Google Scholar] [CrossRef] [PubMed]
[3] Russi, M., Valeri, R., Marson, D., Danielli, C., Felluga, F., Tintaru, A., et al. (2023) Some Things Old, New and Borrowed: Delivery of Dabrafenib and Vemurafenib to Melanoma Cells via Self-Assembled Nanomicelles Based on an Amphiphilic Dendrimer. European Journal of Pharmaceutical Sciences, 180, Article 106311. [Google Scholar] [CrossRef] [PubMed]
[4] Lee, C., Thomas, C.M. and Ng, K.E. (2017) An Overview of the Changing Landscape of Treatment for Advanced Melanoma. Pharmacotherapy: The Journal of Human Pharmacology and Drug Therapy, 37, 319-333. [Google Scholar] [CrossRef] [PubMed]
[5] Cordeiro, A.P., Feuser, P.E., Figueiredo, P.G., da Cunha, E.S., Martinez, G.R., Machado-de-Ávila, R.A., et al. (2021) In vitro Synergic Activity of Diethyldithiocarbamate and 4-Nitrochalcone Loaded in Beeswax Nanoparticles against Melanoma (B16F10) Cells. Materials Science and Engineering: C, 120, Article 111651. [Google Scholar] [CrossRef] [PubMed]
[6] Curran, M.A., Kim, M., Montalvo, W., Al-Shamkhani, A. and Allison, J.P. (2011) Combination CTLA-4 Blockade and 4-1BB Activation Enhances Tumor Rejection by Increasing T-Cell Infiltration, Proliferation, and Cytokine Production. PLOS ONE, 6, e19499. [Google Scholar] [CrossRef] [PubMed]
[7] Koller, K.M., Mackley, H.B., Liu, J., Wagner, H., Talamo, G., Schell, T.D., et al. (2016) Improved Survival and Complete Response Rates in Patients with Advanced Melanoma Treated with Concurrent Ipilimumab and Radiotherapy versus Ipilimumab Alone. Cancer Biology & Therapy, 18, 36-42. [Google Scholar] [CrossRef] [PubMed]
[8] Wolchok, J.D., Chiarion-Sileni, V., Gonzalez, R., Grob, J., Rutkowski, P., Lao, C.D., et al. (2022) Long-Term Outcomes with Nivolumab Plus Ipilimumab or Nivolumab Alone versus Ipilimumab in Patients with Advanced Melanoma. Journal of Clinical Oncology, 40, 127-137. [Google Scholar] [CrossRef] [PubMed]
[9] Da, X., Li, Z., Huang, X., He, Z., Yu, Y., Tian, T., et al. (2023) AGGF1 Therapy Inhibits Thoracic Aortic Aneurysms by Enhancing Integrin α7-Mediated Inhibition of TGF-β1 Maturation and ERK1/2 Signaling. Nature Communications, 14, Article No. 2265. [Google Scholar] [CrossRef] [PubMed]
[10] Zhang, T., Yao, Y., Wang, J., Li, Y., He, P., Pasupuleti, V., et al. (2016) Haploinsufficiency of Klippel-Trenaunay Syndrome Gene Aggf1 Inhibits Developmental and Pathological Angiogenesis by Inactivating PI3K and AKT and Disrupts Vascular Integrity by Activating VE-Cadherin. Human Molecular Genetics, 25, 5094-5110. [Google Scholar] [CrossRef] [PubMed]
[11] He, Z., Song, Q., Yu, Y., Liu, F., Zhao, J., Un, W., et al. (2023) Protein Therapy of Skeletal Muscle Atrophy and Mechanism by Angiogenic Factor Aggf1. Journal of Cachexia, Sarcopenia and Muscle, 14, 978-991. [Google Scholar] [CrossRef] [PubMed]
[12] LaFargue, C.J., Amero, P., Noh, K., Mangala, L.S., Wen, Y., Bayraktar, E., et al. (2023) Overcoming Adaptive Resistance to Anti-VEGF Therapy by Targeting CD5L. Nature Communications, 14, Article No. 2407. [Google Scholar] [CrossRef] [PubMed]
[13] Yu, Y., Li, Y., Peng, H., Song, Q., Da, X., Li, H., et al. (2022) Angiogenic Factor AGGF1 Blocks Neointimal Formation after Vascular Injury via Interaction with Integrin Α7 on Vascular Smooth Muscle Cells. Journal of Biological Chemistry, 298, Article 101759. [Google Scholar] [CrossRef] [PubMed]
[14] Park, B.Y., Grisham, R.N., den Hollander, B., Thapi, D., Berman, T., de Stanchina, E., et al. (2016) Tumor Inhibition by Enzalutamide in a Xenograft Model of Ovarian Cancer. Cancer Investigation, 34, 517-520. [Google Scholar] [CrossRef] [PubMed]
[15] Mao, L., Qi, Z., Zhang, L., Guo, J. and Si, L. (2021) Immunotherapy in Acral and Mucosal Melanoma: Current Status and Future Directions. Frontiers in Immunology, 12, Article 680407. [Google Scholar] [CrossRef] [PubMed]
[16] Willsmore, Z.N., Coumbe, B.G.T., Crescioli, S., Reci, S., Gupta, A., Harris, R.J., et al. (2021) Combined Anti‐PD‐1 and Anti‐CTLA‐4 Checkpoint Blockade: Treatment of Melanoma and Immune Mechanisms of Action. European Journal of Immunology, 51, 544-556. [Google Scholar] [CrossRef] [PubMed]
[17] Yancopoulos, G.D., Davis, S., Gale, N.W., Rudge, J.S., Wiegand, S.J. and Holash, J. (2000) Vascular-Specific Growth Factors and Blood Vessel Formation. Nature, 407, 242-248. [Google Scholar] [CrossRef] [PubMed]
[18] Carmeliet, P. and Jain, R.K. (2011) Molecular Mechanisms and Clinical Applications of Angiogenesis. Nature, 473, 298-307. [Google Scholar] [CrossRef] [PubMed]
[19] Ferrara, N., Hillan, K.J. and Novotny, W. (2005) Bevacizumab (Avastin), a Humanized Anti-VEGF Monoclonal Antibody for Cancer Therapy. Biochemical and Biophysical Research Communications, 333, 328-335. [Google Scholar] [CrossRef] [PubMed]
[20] Krämer, I. and Lipp, H.-P. (2007) Bevacizumab, a Humanized Anti-Angiogenic Monoclonal Antibody for the Treatment of Colorectal Cancer. Journal of Clinical Pharmacy and Therapeutics, 32, 1-14. [Google Scholar] [CrossRef] [PubMed]
[21] Jain, R.K., Duda, D.G., Clark, J.W. and Loeffler, J.S. (2006) Lessons from Phase III Clinical Trials on Anti-VEGF Therapy for Cancer. Nature Clinical Practice Oncology, 3, 24-40. [Google Scholar] [CrossRef] [PubMed]
[22] Wei, H., Zhao, L., Li, W., Fan, K., Qian, W., Hou, S., et al. (2013) Combinatorial PD-1 Blockade and CD137 Activation Has Therapeutic Efficacy in Murine Cancer Models and Synergizes with Cisplatin. PLOS ONE, 8, e84927. [Google Scholar] [CrossRef] [PubMed]
[23] Selby, M.J., Engelhardt, J.J., Quigley, M., Henning, K.A., Chen, T., Srinivasan, M., et al. (2013) Anti-CTLA-4 Antibodies of IgG2a Isotype Enhance Antitumor Activity through Reduction of Intratumoral Regulatory T Cells. Cancer Immunology Research, 1, 32-42. [Google Scholar] [CrossRef] [PubMed]
[24] La Porta, S., Roth, L., Singhal, M., Mogler, C., Spegg, C., Schieb, B., et al. (2018) Endothelial Tie1-Mediated Angiogenesis and Vascular Abnormalization Promote Tumor Progression and Metastasis. Journal of Clinical Investigation, 128, 834-845. [Google Scholar] [CrossRef] [PubMed]
[25] Zhang, Y., Du, X., Liu, M., Tang, F., Zhang, P., Ai, C., et al. (2019) Hijacking Antibody-Induced CTLA-4 Lysosomal Degradation for Safer and More Effective Cancer Immunotherapy. Cell Research, 29, 609-627. [Google Scholar] [CrossRef] [PubMed]

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