Decoding Epileptic Brain Activity: Advancing Human Disease-on-a-Chip Models for Pharmacological Intervention

About the Project

SLC6A1 encodes GAT-1, a voltage-dependent gamma-aminobutyric acid (GABA) transporter responsible for the reuptake of GABA from the synaptic cleft. As the primary inhibitory neurotransmitter in the brain, GABA plays a critical role in regulating neuronal excitability and maintaining the balance between excitation and inhibition. Disruption of this balance, particularly due to impaired GABA reuptake, can result in neuronal hyperexcitability and seizures.

SLC6A1-related neurodevelopmental disorder is characterized by a spectrum of symptoms, including mild to severe developmental delay, intellectual disability, epilepsy, and movement disorders.

Project Summary

This project aims to investigate how single-point mutations in SLC6A1 contribute to abnormal neuronal development and activity, using a combination of clinical insight and cutting-edge human stem cell technologies. Induced pluripotent stem cells (iPSCs) derived from three patients carrying distinct SLC6A1 mutations will be used to generate both neurons and astrocytes. These cell types will be co-cultured to explore how their interactions influence disease phenotypes.

In parallel, patient-derived iPSCs will be used to generate cerebral organoids; three-dimensional brain-like structures to model epileptic brain features more accurately. We will examine neuronal excitability and network dynamics in these models using advanced electrophysiological tools such as patch-clamp recordings and multielectrode arrays (MEAs). Additionally, transcriptomic and proteomic analyses will be performed to identify dysregulated molecular pathways and potential therapeutic targets to reverse aberrant neural function.

This research will provide valuable insights into the molecular mechanisms underpinning SLC6A1-related disorders and may inform the development of targeted therapies for epilepsy and related conditions.

Aim

To elucidate the molecular and functional mechanisms underlying neuronal dysfunction in epilepsy patients with SLC6A1 mutations.

Objectives

• Identify dysregulated pathways in neurons derived from SLC6A1 patient iPSCs co-cultured with either patient- or control-derived astrocytes.
• Characterize molecular and cellular abnormalities in cerebral organoids derived from SLC6A1 patient iPSCs compared to healthy controls.
• Investigate electrophysiological and network-level abnormalities in SLC6A1 patient-derived models.
• Evaluate potential therapeutic targets for rescuing abnormal neuronal functions in SLC6A1 models.

Research Training

This project offers a unique opportunity for comprehensive training in neural stem cell biology. The appointed student will learn to culture and differentiate human iPSCs into neurons and astrocytes and generate 3D cerebral organoids.

Techniques to be acquired:

• Cell Biology: Imaging, immunocytochemistry, Western blotting
• Genetics & Molecular Biology: PCR, RT-qPCR, cloning, RNA sequencing
• Functional Assays: Calcium imaging, whole-cell patch-clamp, MEA recordings
• Omics Approaches: Transcriptomics and proteomics

Generic skills training: The student will have access to the Newcastle University Learning Hub for tailored training in research skills, scientific writing, and data analysis. They will also benefit from attending seminars and collaborative events within the Faculty of Medical Sciences. The successful applicant will join a multidisciplinary research environment involving experts from pharmacy, neuroscience, psychology, and industry partners.

Candidate Requirements

Applicants should hold or expect to obtain a First or Upper Second Class Honours degree (2:1 or above) in a relevant subject such as neuroscience, biomedical sciences, or a related field.

Funding Notes

This project is suitable for self-funded students or students with third-party sponsorship. There is no dedicated funding from the university for this project. The student will be expected to provide funding for tuition fees and living expenses. UK students may be able to apply for a Doctoral Loan from Student Finance for financial support. Some students may be eligible to apply for supplemental funding.

Details about the tuition fees and a supplemental funding search tool are available on our website: View Website