| 研究課題/領域番号 |
22K16375
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| 研究機関 | 熊本大学 |
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研究代表者 |
ELSAYED.NASSER HESHAM 熊本大学, ヒトレトロウイルス学共同研究センター, 特別研究員 (20868020)
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| 研究期間 (年度) |
2022-04-01 – 2025-03-31
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| キーワード | HIV-1 restriction |
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| 研究実績の概要 |
Initial goal is to clarify the APOBEC3 (A3)-mediated restriction of HIV-1 in THP-1 cells, as a model for HIV-1-infected myeloid cells. We created A3F and A3F/A3G-null THP-1 clones using CRISPR approach. HIV-1 Vif degrades A3 proteins in order to replicate efficiently in HIV-1-infected CD4 T cells.In THP-1 cells, disrupted A3F and A3F/A3G proteins fully restored infectivity of Vif-deficient HIV-1. Also, disruption of all A3 proteins (A3-null THP-1) also resulted in restoration of infectivity of Vif-deficient HIV-1. However, we found that endogenous A3H is not involved in HIV-1 restriction in THP-1 cells.
Furthermore, we concluded that restriction of HIV-1 infection by A3 proteins is mediated through a deaminase-dependent (G-to-A mutations) and deaminase-independent mechanisms (accumulation of reverse transcription products). Collectively, our results indicate that A3 proteins are primary target of HIV-1 Vif to be degraded, in order to secure efficient HIV-1 replication in THP-1 cells.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
We successfully established the engineered THP-1 clones (namely A3F, A3F/A3G, and A3 null subclones). Accordingly, infection studies, sequencing, mutation analysis, and late reverse transcription products quantification run smoothly in the appropriate time.
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| 今後の研究の推進方策 |
As we characterized A3-mediated restriction of HIV-1 infection in THP-1 cells, and clarified its underlying mechanisms, we already started to establish THP-1 cells under inflammatory-like condition (IFN treatment), to further evaluate contribution of A3 proteins in HIV-1 restriction under such a condition.
Additionally, we are trying to establish and characterize iPS-ML cell model, which will be utilized to test A3-mediated restriction of HIV-1 infection simulating developmental and physiological conditions in infected patients.
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