# RNA-based regulation of TDP-43 nuclear export in ALS/FTD

> **NIH NIH R01** · JOHNS HOPKINS UNIVERSITY · 2021 · $520,142

## Abstract

TAR DNA-binding protein-43 (TDP-43) is an essential DNA/RNA-binding protein that plays a major role in RNA
processing and stability. Although predominantly nuclear in localization, over 95% of amyotrophic lateral
sclerosis (ALS) cases and up to half of frontotemporal dementia (FTD) cases show mislocalization of TDP-43
to the cytoplasm. Substantial evidence links TDP-43 mislocalization to the pathogenesis of neurodegeneration
in ALS/FTD, and TDP-43 mutations have been identified in subset of familial ALS cases, further supporting a
primary role for TDP-43 disruption in disease. However, the cause of TDP-43 nuclear clearance in ALS/FTD
remains unknown. A major barrier has been lack of understanding of the mechanism of TDP-43 nuclear export.
Until recently, TDP-43 was thought to be a cargo of the nuclear export receptor, exportin-1 (XPO1). However,
multiple recent studies demonstrate that XPO1 does not bind TDP-43 or mediate its export. Alternative export
mechanisms may include passive diffusion from the nucleus, or active co-export with RNA, such as via the
NXF1/TREX mRNA export complex. Our preliminary data show that TDP-43 nuclear export is ATP- and RNA-
dependent, but the energy may be required upstream of the pore, for TDP-43 intranuclear mobility, rather than
translocation across the nuclear pore complex (NPC). Indeed, a permeabilized cell assay suggests that, once
displaced from nuclear RNA, TDP-43 likely diffuses out of the nucleus. Moreover, studies in an NXF1 auxin-
inducible degron (AID) cell line suggest that the bulk mRNA pathway is not required for TDP-43 export. Based
on these data, we hypothesize that ATP-dependent nuclear tethering of TDP-43, most likely to nuclear RNA,
critically dictates the rate at which it diffuses into the cytoplasm. In the proposed studies, we will perform an
arrayed CRISPRi and CRISPRa screen of a targeted library to determine the mechanism of energy-dependent
nuclear tethering of TDP-43. Hits will be validated in nuclear transport and photobleaching assays, and also
evaluated in ALS/FTD patient tissue. We will then combine high content microscopy with nucleocytoplasmic
transport assays, including a newly developed permeabilized cell export assay, to confirm the mechanism of
TDP-43 translocation across the nuclear pore during export. Additional AID cell lines and knockdown studies
will be used to definitively exclude a role for the mRNA export pathway. Finally, we will test the therapeutic
efficacy of a novel approach to promote TDP-43 nuclear retention, identified during our preliminary studies.
Testing will include TDP-43 mutant Drosophila (in collaboration with the Daniela Zarnescu lab), TDP-43 mutant
iNeurons, and mouse sciatic nerve axotomy, to determine if we can promote TDP-43 nuclear retention in vivo,
and whether this strategy will attenuate molecular and neuropathologic features of TDP-43 proteinopathy.
These studies will enable a precise mechanistic understanding of the mechanism and regulation of ...

## Key facts

- **NIH application ID:** 10285495
- **Project number:** 1R01NS123538-01
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Lindsey Renae Hayes
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $520,142
- **Award type:** 1
- **Project period:** 2021-08-01 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10285495

## Citation

> US National Institutes of Health, RePORTER application 10285495, RNA-based regulation of TDP-43 nuclear export in ALS/FTD (1R01NS123538-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10285495. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*