Year: 2025
Type: article
Abstract: BACKGROUND: Alzheimer's disease and related dementias (ADRD) affect 6.2 million people in the U.S., a number that is rapidly increasing due to an aging population, yet no effective cure or treatment exists. TDP-43 pathology, initially identified as a biomarker for ALS and FTD, is now observed in the majority of ADRD patients and is often associated with accelerated cognitive decline. However, the molecular mechanisms by which TDP-43 contributes to ADRD progression remain unclear, largely due to the lack of tools to monitor its aggregation dynamics in living cells. METHODS: To address this gap, we engineered a FRET-based biosensor to detect TDP-43 aggregation. The biosensor was designed using various TDP-43 isoforms (full-length, C-terminal domain, and ΔNLS) fused to different linker constructs. We evaluated biosensor performance under proteostasis stress conditions known to induce TDP-43 aggregation and validated its sensitivity using preformed TDP-43 fibrils. To further explore the progression of TDP-43 pathology, we transduced the biosensor into human iPSC-derived cortical neurons. These neurons were then treated with cellular stressors that mimic aging-related proteostasis dysfunction, cellular stressors that mimic aging-related proteostasis dysfunction, i.e. preformed fibrils and pharmacological drugs. FRET signal growth was monitored over time, and neurons exhibiting high FRET signals were isolated using FACS for downstream RNA-seq analysis. RESULTS: The engineered FRET biosensor successfully detected TDP-43 aggregation in response to proteostasis stressors. Neurons treated with aging-mimicking compounds exhibited an increase in FRET signal over time, indicating progressive TDP-43 aggregation. CONCLUSION: This study presents a novel FRET-based biosensor for tracking TDP-43 aggregation in living neurons. Our approach enables real-time monitoring of TDP-43 pathology and facilitates the identification of molecular pathways disrupted by its accumulation.
Source: Alzheimer s & Dementia
Authors: Han Zhao, Haowei Xu, Junkai Xie, Shichen Wu, Chongli Yuan
Institution: Purdue University West Lafayette
Language: English
Subfield: Neurology
Field: Medicine
Domain: Health Sciences
Open Access status: hybrid
Estimated APC paid: $4,200
OpenAlex ID: W7117163281
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