# Project 2 - Investigation of human neuron models of channelopathy-associated epilepsy

> **NIH NIH U54** · NORTHWESTERN UNIVERSITY · 2021 · $368,420

## Abstract

In Project 2 we will determine the functional consequences of epilepsy-associated ion channel gene variants
using human neurons differentiated from patient-specific induced pluripotent stem cells (iPSCs). We will initially
focus on the SCN2A and KCNQ2 genes, which encode the voltage-gated Na+ (NaV1.2) and K+ (KV7.2)
channels respectively. Mutations in SCN2A and KCNQ2 are responsible for monogenic early onset epileptic
encephalopathy (EE) with overlapping clinical features and diverse severity. Collectively, variants in these two
genes account for ~10% of all mutations identified in genetic epilepsy. The molecular pathogenic mechanisms
responsible for the clinical manifestations of KCNQ2- and SCN2A-related epilepsies remain largely unknown.
More importantly, no targeted therapeutic approach capable of diminishing seizure burden and improving
developmental outcomes exists for these devastating neurological disorders. In Aim 1, we will use patient-
specific cortical neurons to elucidate the functional consequences of epilepsy-associated KCNQ2 and SCN2A
variants. We will specifically examine cortical excitatory and inhibitory neurons derived from existing patient-
specific iPSC lines with pathogenic variants and corresponding isogenic control lines. We will use a
combination of transcriptional profiling (single-cell RNA-sequencing) with electrophysiological approaches
(whole cell patch clamp recording and high-throughput optogenetic recordings) to determine the impact of
mutations on neuronal function and excitability. In Aim 2, we will assess the intrinsic excitability of patient
neurons before and after treatment with NaV channel blockers and KV7 agonists that have clinical efficacy in
the patients from whom the cells were derived. Our goal will be to rank the in vitro effectiveness of drugs in
restoring normal neuron excitability for each genetic variant, and then to correlate the in vitro drug responses
with the clinical responses to AEDs documented for these patients. This project entails a strategic collaboration
between Dr. Kiskinis, whose lab focuses on using stem cell-based approaches to establish models of
neurological disease, and Q-State Biosciences, Inc., which under the scientific leadership of Dr. McManus has
been developing optogenetic technologies to enable high-throughput electrical recordings of human neurons
and drug screening platforms for epilepsy syndromes. This project will work closely with the other Center
teams, including Core A (Variant Prioritization and Curation Core), Project 1 (High-Throughput Functional
Evaluation of Ion Channel Variants) and Project 3 (Development and Investigation of Murine Models of
Channelopathy-associated Epilepsy). Core A is building tools to prioritize variants for experimental evaluation
by the three Center projects. Correlation of findings from Project 2 with those of Projects 1 and 3 will help
determine the reliability and accuracy of iPSC technology to predict in vivo physiology and pharmac...

## Key facts

- **NIH application ID:** 10247557
- **Project number:** 5U54NS108874-04
- **Recipient organization:** NORTHWESTERN UNIVERSITY
- **Principal Investigator:** Evangelos Kiskinis
- **Activity code:** U54 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $368,420
- **Award type:** 5
- **Project period:** 2018-09-30 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10247557

## Citation

> US National Institutes of Health, RePORTER application 10247557, Project 2 - Investigation of human neuron models of channelopathy-associated epilepsy (5U54NS108874-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10247557. Licensed CC0.

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