DOI： 10.1101/2025.11.12.687969

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Publishing type：Research paper (scientific journal)   Publisher：Wiley  

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are progressive neurodegenerative diseases characterised by nuclear TDP‐43 loss. Its hallmark, cryptic exon (CE) splicing, is often masked in bulk tissue analyses by the low abundance of affected neurons. We developed an ultrasensitive RT‐qPCR assay targeting STMN2 CE using one exon–CE junction‐spanning primer and the other within the CE. The design expands the dynamic range sevenfold: TDP‐43 knockdown boosted STMN2 CE levels 1395‐fold in differentiated SH‐SY5Y neurons. Spike‐in tests set detection at 0.16% deficient cells. Crucially, the assay revealed a 42‐fold CE increase in ALS motor cortex, previously missed by conventional primers. This streamlined tool enables precise quantification of TDP‐43 dysfunction and sensitive pharmacodynamic monitoring for future ALS‐FTD therapeutic studies.

Impact statement

Because cryptic‐exon signals are diluted in bulk tissue, we developed a junction‐spanning STMN2 RT‐qPCR with sub‐percent sensitivity. This deployable biomarker will aid ALS/FTD researchers and drug developers by standardizing measurements and enabling sensitive pharmacodynamic monitoring of therapies targeting nuclear TDP‐43 dysfunction.

DOI： 10.1002/1873-3468.70198

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Publishing type：Part of collection (book)   Publisher：Springer Nature Singapore  

DOI： 10.1007/978-981-99-4886-4_13

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

Abstract

In amyotrophic lateral sclerosis (ALS), TAR DNA-binding protein 43 (TDP-43), which is encoded by TARDBP, forms aggregates in the motor cortex. This aggregate formation may be triggered by an increase in the TDP-43 level with aging. However, the amount of TDP-43 is autoregulated by alternative splicing of the TARDBP 3′UTR, and how this autoregulation is affected by aging remains to be elucidated. We found that DNA demethylation in the autoregulatory region in the TARDBP 3′UTR reduced alternative splicing and increased TARDBP mRNA expression. Furthermore, in the human motor cortex, we found that this region was demethylated with aging, resulting in increased expression of TARDBP mRNA. The acceleration of DNA demethylation in the motor cortex was associated with the age of ALS onset. In summary, the dysregulation of TDP-43 autoregulation by age-related DNA demethylation in the motor cortex may explain the contribution of aging and motor system selectivity in ALS.

DOI： 10.1038/s42003-021-02621-0

PubMed

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Other Link： https://www.nature.com/articles/s42003-021-02621-0

Language：English   Publisher：(一社)日本認知症学会  

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Total pages：xi, 402p   Language：Japanese

CiNii Books

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J-GLOBAL

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J-GLOBAL

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J-GLOBAL

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Authorship：Lead author   Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)  

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J-GLOBAL

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