A quantitative cell-based reporter links TDP-43 aggregation and dysfunction to define pathogenic mechanisms.

Overview

1. Edward Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, Missouri, United States of America
2. Department of Neurology, Washington University in St. Louis, St. Louis, Missouri, United States of America
3. Center for Alzheimer’s and Neurodegenerative Diseases, Peter O’Donnell Jr. Brain Institute, UT Southwestern Medical Center, Dallas, Texas, United States of America
4. Department of Pathology and Immunology, Washington University in St. Louis School of Medicine, St. Louis, Missouri, United States of America
5. Agilent Technologies, Santa Clara, California, United States of America

Abstract

TDP-43 pathology is a hallmark of fatal neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), and limbic-predominant age-related TDP-43-encephalopathy (LATE). In affected patients, cytoplasmic TDP-43 aggregates are accompanied by disruption of its normal nuclear localization and function. Because TDP-43 is an RNA binding protein that controls transcript processing, including repression of cryptic exon splicing, its loss leads to dysregulation of gene expression. Despite its central significance in disease, the connection between TDP-43 aggregation and dysfunction remains poorly understood, and models to study the underlying mechanisms are limited. Here, we characterize a robust and quantitative cell-based reporter that captures both aggregation and the resulting loss of function. Using this human biosensor cell line, we show that aggregation initiated by prion-like seeding drives progressive depletion of nuclear TDP-43 and induces signature features of diminished TDP-43 activity, such as increased DNA damage and activation of cryptic exon splicing. We find that aggregate seeding also induces cryptic exon splicing in human neurons implying that this pathological link extends to disease-relevant models. The seeding model provides a platform for dissecting mechanisms that underlie TDP-43 pathology and for identifying factors that modulate the aggregation-to-dysfunction transition. Our data shows that aggregate seeding impacts TDP-43 autoregulation, initiating a toxic feed-forward mechanism that disrupts TDP-43 homeostasis. Furthermore, reducing ataxin-2 levels decreases aggregation and restores TDP-43 activity. Together, these findings reveal a molecularly guided strategy to directly impact TDP-43 activity by decreasing its misfolding and aggregation, highlighting approaches to prevent TDP-43 dysfunction and mitigate toxicity under pathological conditions.

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Data

Datasets cited

• doi:10.17632/yh788t3bxy.1, at the source; found in “Data Availability”

Data Availability

All relevant data can be found within the paper and Supporting information flies. Flow cytometry data are available in Mendeley Data: V1, doi: https://doi.org/10.17632/yh788t3bxy.1.

Reproduced under the paper's license (CC BY), from the paper cited above.

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Version 1, 30 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 11 authors, 12 MeSH terms, 9 funders, 79 references, 1 integrity notice.

Cite

This paper

Mamede, L. D., Hu, M., Vaquer-Alicea, J., Titus, A. R., Passos, P. M., Lantelme, E., French, R. L., Kirschner, P. A., Diamond, M. I., Miller, T. M., & Ayala, Y. M. (2026). A quantitative cell-based reporter links TDP-43 aggregation and dysfunction to define pathogenic mechanisms. PLoS biology, 24(3), e3003662. https://doi.org/10.1371/journal.pbio.3003662

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