# Investigating the Contribution of ALS/FTD-Associated Mutations in the NEK1 Kinase to Disease Pathophysiology

> **NIH NIH R01** · NORTHWESTERN UNIVERSITY · 2023 · $766,016

## Abstract

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease that is characterized by
a progressive inability to control muscle movement. ALS patients are often comorbid with frontotemporal
dementia (FTD), also known as ALS/FTD. The clinical manifestation of ALS is mediated by the selective
dysfunction and degeneration of upper and lower motor neurons (MNs) that connect the CNS to the musculature.
The overwhelming majority of ALS is sporadic in nature, while 10% of patients suffer from familial forms of
disease, which have enabled the identification of causative genetic variants. ALS can be caused by mutations
in genes that encode proteins involved in diverse cellular functions ranging from RNA metabolism, proteostasis
and cytoskeletal homeostasis. Recent genetic studies have highlighted NIMA-related kinase 1 (NEK1) as a major
genetic contributor to ALS. Loss-of-function genetic variants in NEK1 confer susceptibility to ALS in as many as
2% of all cases. The specific role and function of NEK1 in the CNS remains unresolved. What also remains
elusive is the cellular mechanisms that lead to mutant NEK1 ALS pathophysiology. In the present study, we will
use NEK1 cellular models, induced pluripotent stem cell (iPSC) patient-derived MNs, in vivo Drosophila models
and ALS-NEK1 postmortem patient CNS tissue to: a) determine the mechanisms by which ALS-associated
mutations impair MN function; b) characterize the physiological substrates for NEK1-dependent phosphorylation;
and, c) validate the contribution of these changes towards neuropathology in ALS. We will test the hypothesis
that NEK1 mutations cause neurotoxicity by disrupting the regulatory role of the kinase on cellular pathways that
are essential for MN function. In preliminary experiments, we found that NEK1-deficient iPSC-derived MNs
exhibit disrupted microtubule (MT) dynamics and impaired nuclear import. In Aim 1 we will determine whether
these defects are relevant in the context of an extensive set of nonsense and missense ALS-associated NEK1
variants. In preliminary experiments, we found that NEK1 interactors are enriched for function in the MT
cytoskeleton and nuclear import and that reduction of NEK1 levels results in differential expression of proteins
involved in these pathways. In Aim 2 we will determine the physiological substrates for NEK1 phosphorylation
in MNs by conducting phosphoproteomic mass spectrometry analysis and interrogating the functional effects of
differential phosphorylation. In preliminary experiments we identified Niki as the closest Drosophila homologue
of NEK1 and using RNAi lines we found that it is essential for motor function and survival. In Aim 3 we will
determine the function of NEK1 in the intact nervous system of flies and validate our findings on the effects of
the cellular models in vivo. Our studies will shed light into the cellular mechanisms that are compromised by
mutant NEK1 in neurons and will likely uncover potential therapeutic t...

## Key facts

- **NIH application ID:** 10753020
- **Project number:** 1R01NS134166-01
- **Recipient organization:** NORTHWESTERN UNIVERSITY
- **Principal Investigator:** Evangelos Kiskinis
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $766,016
- **Award type:** 1
- **Project period:** 2023-07-15 → 2028-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10753020

## Citation

> US National Institutes of Health, RePORTER application 10753020, Investigating the Contribution of ALS/FTD-Associated Mutations in the NEK1 Kinase to Disease Pathophysiology (1R01NS134166-01). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10753020. Licensed CC0.

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