2022 Fiscal Year Research-status Report
mRNA therapy for the treatment of autoimmune diseases that produces chimeric antigen receptor regulatory T cells in vivo
| Project/Area Number |
22K19541
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Cabral Horacio 東京大学, 大学院工学系研究科(工学部), 准教授 (10533911)
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| Co-Investigator(Kenkyū-buntansha) |
持田 祐希 公益財団法人川崎市産業振興財団(ナノ医療イノベーションセンター), ナノ医療イノベーションセンター, 主任研究員 (60739134)
松元 亮 東京医科歯科大学, 生体材料工学研究所, 准教授 (70436541)
宮崎 拓也 地方独立行政法人神奈川県立産業技術総合研究所, 「貼るだけ人工膵臓」プロジェクト (松元P), 非常勤研究員 (80844779)
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| Project Period (FY) |
2022-06-30 – 2024-03-31
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| Keywords | mRNA / polymeric micelles / Treg / Foxp3 |
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| Outline of Annual Research Achievements |
We are developing polymeric micelles to facilitate the delivery of mRNA to T cells to generate Treg cells for diabetes therapy. Initially, we synthesized Azide-PEG-poly(glycidyl tyrosinate) (N3-PEG-PGTyr) as our starting material. The micelles were then prepared by combining mRNA with N3-PEG-PGTyr at various mixing ratios in a buffer solution. We determined the optimal ratio based on the micelle size, and their stability against heparin and serum. We also generated anti-CD3 antibody fragments by enzymatically digesting whole antibodies. These fragments were then conjugated to the micelles using click chemistry, resulting in 2-3 fragments on the micelles' surface. The fragments on the micelles enhanced mRNA translation in T cells.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The smooth progress of the project was given by the successful synthesis of N3-PEG-PGTyr for consistent and reliable micelle formulation, the proper assembly of the mRNA-loaded micelles and the installation of the antibody fragments on their surface. Moreover, the antibody fragment-installed mRNA-loaded micelles worked in vitro by inducing high protein expression in T cell. These results support the design of the particles and encourage further development in vivo for the delivery of mRNA encoding Foxp3 to generate Tregs.
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| Strategy for Future Research Activity |
The next step will involve conducting in vivo studies to evaluate the performance and effectiveness of the antibody fragment-installed mRNA-loaded micelles. We will test the delivery of mRNAs encoding luciferase as surrogate and Foxp3 to generate Tregs in mice. We will assess the micelles ability to induce high protein expression in vivo. Based on the in vivo data, further optimization and refinement of the micelle formulation will be pursued by fine-tuning the ligand-density and enhancing the stability. In addition, we will test the ability of the Tregs generated by the micelles to protect the pancreas of diabetic mice. We will do this by checking the glucose levels in blood and by histological analysis of pancreatic tissues.
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| Causes of Carryover |
The following consumables are necessary: mRNA preparation kit to make mRNA encoding Foxp3, chemicals for polymer synthesis, antibodies, appropriate animal models, such as mice or diabetic mice. Moreover, we will need budget to cover trip to conferences and animal housing facilities.
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