| Project/Area Number |
20K09819
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 56060:Ophthalmology-related
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| Research Institution | Okinawa Institute of Science and Technology Graduate University |
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Principal Investigator |
CHIANG WeiChieh 沖縄科学技術大学院大学, 神経発生ユニット, スタッフサイエンティスト (70867754)
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| Project Period (FY) |
2020-04-01 – 2024-03-31
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| Project Status |
Granted (Fiscal Year 2021)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2023: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000)
Fiscal Year 2022: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | ER stress / ER proteostasis / retina / Retinal Degeneration / Photoreceptor Cells / Protein Misfolding / ER Stress / Stress Response |
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| Outline of Research at the Start |
Endoplasmic reticulum (ER) is an important cellular organelle that is essential for the proper folding of secretory and membrane protein. Many retinal degenerative diseases arise from the imbalance of ER proteostasis. To date, the links between such imbalance and retinal cell death is still largely unclear.
Recently, ER membrane protein complex (EMC) was found to be a key ER factor in membrane protein synthesis and trafficking. In this study, I aim to decipher how EMC regulates ER proteostasis in the retina and the molecular mechanism of retinal degeneration caused by EMC-dysfunction.
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| Outline of Annual Research Achievements |
ER membrane protein complex (EMC) is an important factor for ER homeostasis maintenance and membrane protein biogenesis. Using an EMC-deficient zebrafish model, I found that mutant fish display various retinal phenotypes, including the loss of photoreceptor cells and lack of visual function. Additionally, an EMC-deficient zebrafish carrying a reporter gene that monitor UPR activation in real time, I found that UPR is activated as early as 3 days post fertilization (dpf). This finding is further confirmed with bulk RNA sequencing analysis and qPCR analysis. These data suggest that ER stress and UPR may play a critical role in regulating retinal degeneration associated with EMC-dysfunction.
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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
To assess the global stress response induced by EMC-deficiency, I have performed the bulk RNA sequencing analysis of the EMC-deficient zebrafish eyes, followed by validation using qPCR analysis . I found that ER stress response/UPR is strongly activated up to 6 dpf. Interestingly, UPR activity starts to decline at 7 dpf with some part of PERK pathway still activated. I also identified genes, involved in cell death mechanisms, are up- or down-regulated. These data will provide insights on how ER stress regulates retinal degeneration associated with EMC-deficiency.
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| Strategy for Future Research Activity |
Since I have identified genes that may be involved in EMC-deficiency associated retinal degeneration, I am generating over-expression or CRISPR/Cas9 KO zebrafish of these genes to investigate how these genes modulate retinal cell death in the EMC-deficient fish. I will also perform gene expression analysis to determine how these genes are regulated by UPR. These studies will provide novel evidence on how UPR facilitate cell death mechanism in the retinas that suffer from the disruption of ER proteostasis.
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