Project/Area Number |
22K16374
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Research Category |
Grant-in-Aid for Early-Career Scientists
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Allocation Type | Multi-year Fund |
Review Section |
Basic Section 54030:Infectious disease medicine-related
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Research Institution | Kumamoto University |
Principal Investigator |
BARABONA GODFREY 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 (40906674)
|
Project Period (FY) |
2022-04-01 – 2024-03-31
|
Project Status |
Granted (Fiscal Year 2022)
|
Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2023: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2022: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
|
Keywords | Extracellular vesicle / HIV / Inflammation / MicroRNA |
Outline of Research at the Start |
This study will analyze circulating microRNAs that are delivered by extracellular vesicles and determine their role in systemic immune dysfunction and chronic inflammation seen in HIV infection. The central hypothesis is that, in HIV infection, microRNAs are dysregulated and play a crucial role in the persistent immune dysfunction and chronic inflammation.
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Outline of Annual Research Achievements |
Our project had two main specific aims: (1) to identify circulating miRNAs that are altered in HIV-infected individuals, and (2) to investigate whether altered miRNAs in extracellular vesicles from HIV patients regulate cytokine expression ex vivo. Over the past year of project implementation, we have collected three sets of plasma samples: from HIV-infected individuals before and after treatment, and from HIV-uninfected individuals. Using these samples, we have demonstrated a differential relative expression of miRNAs in plasma extracellular vesicles (EVs). Notably, our preliminary data suggest a trend of downregulation of EV miRNAs in the untreated HIV-infected group compared to the treated and HIV-uninfected groups. Our extended analysis further indicates that absolute levels of miRNA are lower in HIV-infected untreated individuals compared to treated and uninfected individuals. Additionally, we found that plasma miRNAs were more abundant compared to miRNAs in EVs but followed a similar trend to that of EVs. Given that absolute levels of miRNAs seem to be low in most individuals, we intend to first demonstrate the effect of EV miRNAs from HIV-infected individuals compared to those from HIV-uninfected individuals in vivo.
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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
We have successfully obtained the plasma samples required for our study and conducted our initial experiments and analysis. However, we have observed that the levels of miRNAs in EVs are generally low, which may limit their physiological significance. However, we acknowledge that this observation is limited to a small set of miRNAs investigated. Therefore, to further explore the potential impact of miRNAs in EVs on inflammation, we plan to investigate their effects on macrophage inflammation state in vivo. This will allow us to demonstrate the plausibility that miRNAs in EVs at physiological levels can influence inflammation.
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Strategy for Future Research Activity |
We intend to use monocyte-derived macrophages to investigate the impact of EV extracted from HIV-infected individuals and compared it to that from HIV-uninfected individuals. We will use cytokine expression levels to detect the impact of the EV content on the inflammation state. Following this series of experiments, we will then use the combination of next-generation sequencing and bioinformatics tools to identify potential microRNA responsible for the modulation of cytokines expressions in macrophages. To confirm the contribution of miRNA cargo in cytokine production, we will conduct another experiment in the presence of specific anti-miRNAs corresponding to the miRNA candidates that are altered in extracellular vesicles of HIV patients. Finally, we will confirm our observation by transfection of mimic miRNA for each miRNA whose expression in extracellular vesicles of HIV infected individuals were altered
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