| 研究課題/領域番号 |
20K22793
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| 研究機関 | 愛知県がんセンター(研究所) |
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研究代表者 |
呉 智聞 愛知県がんセンター(研究所), 腫瘍免疫応答研究分野, リサーチレジデント (80883983)
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| 研究期間 (年度) |
2020-09-11 – 2022-03-31
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| キーワード | CAR-T / metabolism / immunotherapy |
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| 研究実績の概要 |
We have explored metabolic targets associated with T cell functions and identified that CAR-T cells treated with dorsomorphin (DM), a selective AMPK inhibitor, significantly maintained a young memory phenotype and cytokine polyfunctionality in a dose-dependent manner without affecting proliferative capacity compared with untreated group.
DM-treated T cells showed reduced levels of phosphor-Akt as well as phosphor-AMPK. When DM-treated T cells were infused into NSG mice, they showed enhanced persistence compared with the control T cells.
In addition to pharmacologic inhibition, we also tested the effect of genetic knockout (K.O.) and dominant-negative (DN) mutant modification of AMPK subunit (PRKAA1) in CAR-T cells. The result is consistent with DM-treated group in vitro.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
3: やや遅れている
理由
The AMPK pathway is considered to redirect cellular metabolism towards increased catabolism and decreased anabolism through negatively regulating one of the main anabolic pathway mTOR/Akt. Unexpectedly, dorsomorphin-treated T cells also showed reduced levels of phosphor-Akt as well as phosphor-AMPK, suggesting that AMPK inhibition paradoxically suppresses Akt signaling, which may result in supporting memory T cell formation.
We are working on the mechanism according to the phenotype we observed and try to figure it out to catch up the procedure as planned.
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| 今後の研究の推進方策 |
Molecular mechanism on how Dorsomorphin affects memory T cell differentiation will be investigated. We will perform RNA-sequencing analysis to elucidate how the AMPK inhibition affects transcriptional profiles of CAR-T cells.
We will analyze metabolic changes including glycolysis, oxygen consumption, fatty acid oxidation and glutaminolysis in the AMPK-inhibited T cells compared with control T cells. The effect of PRKAA1-KO and DN-PRKAA1 in CAR-T cells will be evaluated in vivo as well.
The persistence and differentiation of CAR-T cells with various treatment will be analyzed.
Next, we will test antitumor efficacy of dorsomorphin-treated CAR-T cells in the solid tumor model. Mice will be regularly injected with dorsomorphin or vehicle and analyzed for tumor progression.
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| 次年度使用額が生じた理由 |
It is necessary to be used for next year because immunodeficient mice (NSG mice) will be purchased to perform in vivo functional analysis of T cells. Meanwhile we will analyze gene expression profiles of T cells by RNA sequencing.
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