2020 Fiscal Year Research-status Report
DOCK11ノックダウンによりcccDNA排除の新規抗HBV治療応用への基礎研究
| Project/Area Number |
18K07966
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| Research Institution | Kanazawa University |
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Principal Investigator |
李 影奕 金沢大学, 医学系, 博士研究員 (70401940)
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| Co-Investigator(Kenkyū-buntansha) |
本多 政夫 金沢大学, 保健学系, 教授 (00272980)
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| Project Period (FY) |
2018-04-01 – 2022-03-31
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| Keywords | cccDNA / DOCK11 / AGAP2 / ARF1 / antivirus therapy |
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| Outline of Annual Research Achievements |
cccDNA is reservoir for viral persistence and a key obstacle for a cure of CHB. A small number of cccDNA-positive hepatocytes might be a source for viral reactivation during immune suppression. We established a new HCC cell line (named KM) in order to elucidate the host factors required for HBV persistence. We identified human DOCK11 as a one of candidate host factors to maintain cccDNA in KM cells. Our data revealed that DOCK11 activated retrograde trafficking of HBV from early endosomes to the TGN and then to the endoplasmic reticulum, thereby avoiding lysosomal degradation. Moreover, DOCK11 partners AGAP2 and ARF1, essential regulators of retrograde trafficking in the Golgi, were identified using LC-Ms/Ms and were involved in HBV trafficking. Furthermore, DOCK11 and AGAP2 might accelerate HBV recycling possibly through the TGN-ER to nucleus pathway. Additionally, in vivo mouse experiments, DOCK11 shRNA decreased the level of cccDNA in C57BL6 mice which were received AAV-HBV1.3 particles. Clinically, DOCK11 levels in the liver of patients with CHB were significantly reduced by ETV treatment, and this reduction correlated with HBs antigen levels. Thus, DOCK11 might be a candidate druggable target for HBV cure.
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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 project is going smoothly according to the experimental plan. Up to now, our data revealed that DOCK11 maintains a persistent HBV infection by selecting a retrograde trafficking route that avoids lysosomal degradation and possible cytosol immune-mediated viral degradation.
1) In the cell experiment in vitro, we summarized that DOCK11 with its partners AGAP2 and ARF1 activated retrograde trafficking of HBV from early endosomes to the TGN and then to the endoplasmic reticulum, thereby avoiding lysosomal degradation.
2) DOCK11 and AGAP2 might accelerate HBV recycling possibly through the TGN-ER to nucleus pathway in the cell experiment in vitro.
3) In vivo mouse experiments, we made the conclusion that DOCK11 shRNA decreased the level of cccDNA in C57BL6 mice which were received AAV-HBV1.3 particles.
4) Clinically, we got the results that DOCK11 levels in the liver of patients with CHB were significantly reduced by ETV treatment, and this reduction correlated with HBs antigen levels.
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| Strategy for Future Research Activity |
1. We have already completed the manuscript and submitted to Hepatology.
2. We will be revising the editor’s comments.
3. we will be presenting our research achievement in JDDW2021 and will be discussing it with other experts in close way.
4. We will be examining the detail mechanism of DOCK11 on HBV recycling in the hepatocyte infected by HBV particles.
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| Causes of Carryover |
1. As COVID-19 epidemic in the world, academic meeting could not be held in last year. In this year, we will be presenting our research achievement in JDDW2021 and will be discussing it with other experts in close way.
2. It will be for articles submitting and revising.
3. It will be utilized for ordering materials which were used to perform the experiments on examining the detail mechanism of DOCK11 on HBV recycling in the hepatocyte infected by HBV particles
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Research Products
(1 results)
