2019 Fiscal Year Research-status Report
Pathologic hematopoietic stem cell determines co-morbid symptoms in autism
| Project/Area Number |
19K16529
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Lin ChiaWen 国立研究開発法人理化学研究所, 脳神経科学研究センター, 訪問研究員 (20730253)
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| Project Period (FY) |
2019-04-01 – 2021-03-31
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| Keywords | autism / comorbid symptom / inflammation / progenitors / dysbiosis |
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| Outline of Annual Research Achievements |
Yolk sac (YS) and aorta gonad mesonephros (AGM) hematopoietic stem cell (HSC) were compared between B6 and BTBR mice. Single cell trascriptome (sc-RNA seq) was analyzed by 10x genomics. YS and AGM samples were enriched for cKit+ and CD31+ cells, respectively, to increase the recovery rate of HSC.
To access the mechanism of dysbiosis in autism, co-embryo transfer (co-ET) experiment was done by transfer both B6 and BTBR embryos into the same recipient mice of strain 129. Second, to demonstrate autistic immunity determines specific microbiome, the immunity of B6 was swapped into BTBR' by BMT.
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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
Target cell populations in YS and AGM were enriched by magnetic cell separation strategies. The efficiency of enrichment was confirmed by FACS, however, it was not as high as expected. To increase the number of target HSC, I collected 2 bathes of samples for each genotype for sc-RNA seq and combined them at final analysis step.
Co-embryo transfer experiment was done as proposed. After separating b6 and btbr mice for 3 month (from postnatal day 7), their cecal contents were collected for metagenomic analysis.
For BMT experiment, I first shielded the head from irradiation to observe the effect of autistic immunity on behaviors. However, this made low efficiency of reconstitution. By total body irradiation instead, B6 mice swapped for BTBR immunity were further analyzed.
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| Strategy for Future Research Activity |
To identify the molecular mechanism underlying pathological HSC development, sc-RNA seq analysis by Seurat package will be run first to identify the target population and to extract the molecular difference between b6 and autistic BTBR.
According to the result of metagenomic sequencing, butyrate-producing bacteria were largely reduced in BTBR. To investigate the potential of microbiota manipulation on autistic behavior, pregnant btbr will be co-housed with b6 mice to change the microbiota and to effect the pups from early developmental stage. On the other hand, butyarte supplement in pregnant BTBR from early gestation will be done to explore the therapeutic strategy for autism treatment.
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Research Products
(1 results)
