2023 Fiscal Year Research-status Report
Exploring the role of adenosine A2A receptors in Schizophrenia using opto-pharmacologically controlled allosteric modulation.
| Project/Area Number |
23K14685
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| Research Institution | University of Tsukuba |
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Principal Investigator |
ROY Koustav 筑波大学, 国際統合睡眠医科学研究機構, 研究員 (30893086)
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| Project Period (FY) |
2023-04-01 – 2026-03-31
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| Keywords | Schizophrenia / A2AR receptor / Nucleus accumbens (NAc) / Optopharmacology |
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| Outline of Annual Research Achievements |
The coexistence of D2R and A2AR receptors led to the idea of modulating D2R by modulating A2AR. Therefore, a selective increase in A2AR activity in the nucleus accumbens (NAc) could alleviate psychotic problems in schizophrenia, which has not yet been investigated. My central hypothesis is that optopharmacology-based induction of hyperadenosinergic activity by A2AR in the NAc can alleviate psychotic problems in a schizophrenia mouse model. Using our newly developed opto-allosteric modulator optoA2AR PAM, recently published in Nature Communications (Roy et al., Nature Communications 2024), we were able to successfully manipulate the SWS. We proved that adenosine derived from neuron and glia both are responsible A2AR mediated sleep induction in NAc.
We performed behavioral experiments in psychotic mice (MAP6KO) and found MAP6KO mice spend significantly high time in central zone in open filed test without increase of total distance travel which indicate the anxiety behaviour of MAP6KO mice. After injection of the A2ARPAM salt compound, in which we observed a significant decrease in exploration rate. The behavioral experiments will continue to find out whether hyperadenosinergic activity by A2ARPAM can enhance the cognitive performance of MAP6KO mice or not. I am also performing a spatial transcriptome on MAP6KO mice in different states (sleep, wakefulness). Moreover, till now I have documented opto allosteric control of SWS in NAc (Roy et al., Nature Communi. 2024) with identify of selective genes which probably reasonable for sleep alteration in psychotic mice.
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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
In my project proposal, I've outlined the use of the 10X Chromium Single Cell Gene Expression Platform, focusing on single cell/nucleus transcriptome analysis. Currently, I'm actively engaged in executing this aspect of the project. However, I've identified an opportunity to enhance our understanding by incorporating spatial transcriptomic, which was not originally included in the proposal. Spatial transcriptomic offers a more detailed perspective on cellular localization within tissues. By using this technique, we can dissect brain tissue from various groups, each with a thickness of 10 microns, onto Visium slides provided by 10X Genomics. After permeabilization of the tissue recommended by 10X Visium protocol the RNA is bind with the spatial barcodes and unique molecular identified (UMI) on the slides, so that we can identify the spatial information and RNA from different cells.
This extension allows us to discern which areas of the striatum are predominantly affected in MAP6KO mice. Furthermore, it enables identification of specific cell types associated with particular locations within the tissue. This addition promises to enrich the depth and breadth of our research, potentially yielding novel insights into the pathology of psychotic/schizophrenic (MAP6KO) mice. Basicall we include one more technique, so my next experiments will be bit delay.
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| Strategy for Future Research Activity |
I am currently working on a transcriptome analysis (Spatial transcriptomic) of psychotic mice (MAP6KO) and trying to find out the key genes that are mainly responsible for sleep alteration in psychotic mice. In addition, I will investigate the transcriptomics of individual nuclei of the NAc in different states of psychotic mice. The reason for the altered sleep in psychotic mice is still unknown. I am trying to identify the genes from our transcriptome data and through spatial transcriptomics we will map the spatial information of the cells responsible for the poor sleep quality in psychotic mice.
We have previously documented that A2ARPAM injection improves SWS sleep time in psychotic mice (Lin and Roy et al., Front.in Pharmaco. 2023). In the final phase, we will perform optoallosteric modulation of A2AR in the NAc to improve SWS in psychotic mice. Overall single cell transcriptomics, Spatial transcriptomics, opto pharmacologic manipulation and behavior tests will be conducted to investigate our objectives.
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| Causes of Carryover |
The entire project was designed to conduct in three years in order to achieve all the goals. I am currently analyzing the spatial transcriptome data achieved from different condition, whereby, according to my plan, single cell nucleus needs to be performed from different condition of psychotic mice. Performing the transcriptome (10X genomics kit) itself is very expensive, as well as sequencing the cDNA library cost is also higher side. In addition, I perform the opto-pharmacological sleep manipulation, for which I have to buy various consumables with optical fibers and EEG electrodes.
In very brief, I need to use the grant in next financial year to continue my experiments and to achieve my next objectives.
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