2018 Fiscal Year Research-status Report
Studying and targeting a proto-oncogenic Internal Ribosome Entry Site (IRES) in p53 mRNA that is activated by the most frequent mutations in cancer
| Project/Area Number |
18K07229
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | p53 / p53 mRNA / mRNA translation / IRES / Cancer mutation / p53 isoform / delta160p53 / p53 protooncogene |
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| Outline of Annual Research Achievements |
In one year we have accomplished already more than half of the project. It is going extremely well. We should conclude this project within this fiscal year.
For task 1 we have defined the IRES region and done detailed chemical probing with RNases and DMS.
For task 2 we have already tested and identified mutations that strongly induce IRES function. Interestingly only gain-of-function mutations but not loss-of-function mutations in this region activate the IRES. We will be submitting the paper soon. We are now also testing how these mutations affect IRES structure; this could help us improve the design for a drug that could inhibit the IRES and thus the expression of p53 oncogenic isoform D160p53.
For task 3 we have already seen that antisense oligos that target and inactivate D160p53 IRES induce apoptosis in stressed cells and restrains cell proliferation. We are now already testing IRES function in soft agar colony formation assays and will soon start testing the xenograft model.
For task 4 we already have the constructs and the stable cell lines established and are ready to initiate the screenings which should take a couple of months.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
Task 1 is almost concluded and other tasks are well underway (more than half done).
For task 1 we have defined the IRES region and done detailed chemical probing with RNases and DMS.
For task 2 we have already tested and identified mutations that strongly induce IRES function. Interestingly only gain-of-function mutations but not loss-of-function mutations in this region activate the IRES. We will be submitting the paper soon. We are now also testing how these mutations affect IRES structure; this could help us improve the design for a drug that could inhibit the IRES and thus the expression of p53 oncogenic isoform D160p53.
For task 3 we have already seen that antisense oligos that target and inactivate D160p53 IRES induce apoptosis in stressed cells and restrains cell proliferation. We are now already testing IRES function in soft agar colony formation assays and will soon start testing the xenograft model.
For task 4 we already have the constructs and the stable cell lines established and are ready to initiate the screenings which should take a couple of months.
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| Strategy for Future Research Activity |
Everything is going smoothly. All the tasks are well underway, some already concluded and others almost done. Indeed, more than half the project has been completed; we should finish before March 2020. We will continue as planned, without stopping.
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| Causes of Carryover |
Usage plan as predicted.
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