Modulating Neuronal Network Dysfunction in mTORopathies via Peptide-Mediated Inhibition of Rheb/mTOR Signalling

About the Project

Hyperactivation of mammalian target of rapamycin (mTOR) signalling in neurons is associated with aberrant axonal and dendritic connectivity, enlarged soma size, increased cellular stress, reduced myelination, synaptic dysfunction and neuronal hyperexcitability. Inhibiting mTOR with rapamycin can modulate neuronal function and reduces hyperexcitability, but may impair executive and recognition memory, and shows no benefit for neurocognitive or behavioural issues in patients with mTORC1 hyperactivation.

We have previously used patient induced pluripotent stem cells (iPSCs)-derived neurons with Tuberous sclerosis (TSC) mutation to understand the effect of knocking down TSC function (associated with mTORC1 hyperactivity) on neuronal network behaviours (Alsaqati et al., 2021). This research showed that promoting autophagy can rescue the aberrant neuronal network behaviours seen in TSC patient neurons. We have then generated a homozygous TSC stable knockout (KO) human iPSC cell line using CRISPR-Cas9 technology.

We are now planning to use those cells to generate glutamatergic neurons and investigate the effect of a novel approach to inhibiting mTOR activity on neuronal network behaviours, with potential applications for patients with neurological disorders linked to mTORC1 hyperactivation. Specifically, we plan to employ peptide inhibitors to precisely disrupt the RAS/mTOR signalling pathways. This approach will utilise first-in-class peptides, a proprietary class developed by one of the Principal Investigators, Dr. Al Musaimi. These multifunctional peptides will be used to effectively inhibit downstream Rheb signalling. They can be engineered to specifically target neurons in a neuron–astrocyte co-culture system, making them ideal carriers for delivering these inhibitors to neurons in the human brain. In addition, they are capable of crossing cell membranes, have demonstrated a favorable safety profile, and show promise in the treatment of brain injuries.

Aim

To investigate the effects of a small peptide inhibitor of mTORC1 pathways on the functional properties of neurons and astrocytes derived from TSC patient‑specific iPSCs.

Objectives

1. To design and validate a small peptide that selectively targets and inhibits mTORC1.
2. To evaluate the impact of the peptides on mTORC1 activity and its downstream effectors in TSC iPSC-derived neurons and astrocytes.
3. To assess how peptide-mediated modulation of mTORC1 affects functional abnormalities in TSC-derived neural cells, including electrophysiological properties and network behaviours.

This project represents a unique opportunity to gain in-depth training in neural stem cell and peptide synthesis. The appointed student will be trained in culturing human iPS cells and their differentiation into neurons and astrocytes.

Techniques to be used

A variety of cellular, genetics, molecular and functional techniques will be combined, including:

• Cell biology; imaging, Western blotting
• Genetics; PCR and RT-qPCR, cloning
• Functional Assays; calcium imaging, Patch-clamp and Multi-electrode arrays
• Advanced molecular biology techniques; RNA Seq
• Peptide synthesis
• Modification
• Tagging
• Lipidation

The student will gain hands-on experience using advanced instruments, including HPLC, LC-MS, NMR, SEM, and TEM.

Training will also cover effective dissemination of research through publishing articles (research and reviews), participating in conferences, and presenting at research seminars within the NU School of Pharmacy, partner institutes, and other relevant conferences.

Generic skills training is offered through the Newcastle University learning HUB. As well as the specific training detailed above, students will have access to a wide range of seminars and training opportunities in the University. The appointed student will be part of the Faculty of Medical Sciences- Newcastle University. The successful candidate will join the research group at Newcastle University which includes academics and students from pharmacy, dentistry, neuroscience and psychology and they will have opportunities to interact and work with our industry partner/s. Applicant should hold or expect to hold a 2:1 or 1st class degree.

References

• Alsaqati M, Heine VM, Harwood AJ. Pharmacological intervention to restore connectivity deficits of neuronal networks derived from ASD patient iPSC with a TSC2 mutation. Mol Autism. 2020;11(1):80. doi: 10.1186/s13229-020-00391-w
• Al Musaimi O, Morse SV, Lombardi L, Serban S, Basso A, Williams DR. Successful synthesis of a glial-specific blood-brain barrier shuttle peptide following a fragment condensation approach on a solid-phase resin. J Pept Sci. 2023;29(2):e3448. doi: 10.1002/psc.3448
• Coburn F, Nsereko Y, Armstrong A, Al Musaimi O. Peptide inhibitors: Breaking cancer code. Eur J Med Chem. 2025 Nov 5;297:117961. doi: 10.1016/j.ejmech.2025.117961
• Armstrong A, Coburn F, Nsereko Y, Al Musaimi O. Peptide-Drug Conjugates: A New Hope for Cancer. J Pept Sci. 2025;31(8):e70040. doi: 10.1002/psc.70040

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