Discussion

Part 1. DSCR1 regulates AHN by regulating crosstalk between TET1 and miR-124

IV. Tables

Ⅴ. Discussion

DSCR1 leads to delayed neuronal differentiation in the developing neocortex of mice ⁷⁴. However, the precise role of DSCR1 for AHN and dural lymphatics remains unexplored.

In this thesis, we found DSCR1 interacts with TET1 pre-mRNA using the RRM domain, which is sufficient to modulate the splicing of TET1.Previous studies identified that RNA-binding proteins (RBPs) play a critical role in post-transcriptionally regulating gene expression and synthesizing the protein ⁷⁵. Meanwhile, dysfunction or changed levels of RBPs have been associated with various disorders, such as neurological disorders and cancers ^{76, 77}.However, the precise role of DSCR1 in the RNA–protein interactions study has not been fully understood. Thus, it could provide a remarkable research opportunity in the future.

It is also particularly interesting that transcriptomic changes in Wnt signaling pathway respond to DSCR1 in our RNA-seq results. Wnt signaling is required to regulate lymphangiogenesis and differentiation of the lymphatic vascular system ^{78, 79}, implying that the DSCR1 overexpression may improve dural lymphatics by correcting the Wnt pathway. Furthermore, Wnt signaling pathway regulates various cellular and developmental biological processes ^{78, 79}. Therefore, it would be interesting to identify further the role of the DSCR1-Wnt pathway in those processes, including adult hippocampal neurogenesis.

Strikingly, correcting the levels of DSCR1 is sufficient to recover the defects of the adult hippocampal neurogenesis and Aβ pathogenesis in DS and AD mice, resulting in improved cognitive function.

However, overexpressed DSCR1 induces defective adult neurogenesis in the hippocampus, while DSCR1 overexpression alleviates Aβ pathology and improves learning and memory by potentiating the meningeal lymphatic system. Thus, the additional functional study of adult neurogenesis by DSCR1 remains to be investigated in the pathogenesis of AD.

NFAT-calcineurin signaling is one of the main pathways triggered by DSCR1 in the adult and developing CNS ⁸⁰. Even though DSCR1 is the representative regulator of calcineurin, we identified DSCR1 also controls miR-124-TET1 pathway. Therefore, it would be interesting to determine whether DSCR1-mediated calcineurin/NFAT signaling pathway controls the AHN together or independently with miR-124-TET1.

67

ⅤI. Conclusion

This thesis represents two principal conclusions. First, DSCR1 regulates miR-124 expression and AHN by modifying the splicing of TET1. And second, DSCR1 promotes the development of meningeal lymphatics, which lead to an improvement of Aβ pathogenesis including accumulation of Aβ and cognitive defects in 5XFAD mice.

The first of these conclusions support a novel role of DSCR1 in the processes of AHN. We identify that TET1 mediates methylation status in the promoter region of miR-124, which influences its levels. We further found that DSCR1 regulates the TET1 mRNA splicing by competing for U1 and U2 snRNPs of the spliceosome. These findings showed the relationship between DNA methylation and microRNA (miRNA) expression in the AHN, which is supported by our several results. First, we demonstrated that DSCR1 directly interacted with TET1 pre-mRNAs using RRM. Next, DSCR1 knockdown enhanced splicing of TET1 and expression, which was confirmed in the hippocampus of DSCR1 KO mice. Also, altering TET1 expression in DSCR1 null mice could recover AHN defects. Furthermore, correcting a single copy of DSCR1 in Ts65Dn mice fully rehabilitated AHN and learning and memory abnormalities.

Collectively, these results suggest that proper expression of DSCR1 in the hippocampus is required for the normal AHN, subsequently regulating miR-124 and TET1 expression. Therefore, our study increases knowledge of mechanisms about controlling AHN and proposes an unrecognized therapeutic strategy for treating impairment in intellectual functioning in DS.

The second of our principal conclusions demonstrate that DSCR1 potentiates the dural lymphatics and improves lymphatic sewerage. We generated the hybrid mice of DSCR1 TG crossed with AD mice and found not only improved Aβ pathogenesis with enhanced lymphatic sewerage but also advanced cognitive behavior in those mice. Blocking cLVs by ligation surgery in hybrid mice aggravates the Aβ pathogenesis in the dural lymphatics and parenchyma of the brain, showing the reduced cognitive ability.

Also, i.c.v viral deliveries of DSCR1 into the CSF showed improved dural lymphatic drainage and increased learning and memory.Moreover, RNA-seq exhibited a dysregulated transcriptome in mLECs of AD mice, whereas the hybrid mice showed a noticeable transcriptional remodeling.

The dural lymphatic system has critical roles in numerous neurodegenerative diseases ⁸¹, brain injuries

20, and tumors ^{82, 83}, including AD ^{15, 84}. Moreover, it would be meritorious to examine whether DSCR1 could improve age-related atrophy of the meningeal lymphatics with aggravated cognitive dysfunction.

Therefore, DSCR1 would be a possible drug target not only for the therapy of Alzheimer’s disease but also for other several disorders containing dural lymphatics defects.

68

ⅥI. References

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