Job ID: 106506

PhD project – Vector-based targeting of RNA therapeutics to the brain for the development of novel treatments for neurodegenerative disorders

Position: Ph.D. Student

Deadline: 2 April 2023

Employment Start Date: 2 October 2023

Contract Length: 3 years

City: Marseille

Country: France

Institution: Aix Marseille Université

Department: Institute of NeuroPhysiopathology (INP)


The NeuroSchool PhD Program of Aix-Marseille University (France) has launched its annual calls for PhD scholarships for students with a master’s degree in a non-French university.

The following project is one of the 14 proposed projects. Not all proposed projects will be funded, check our website for details.


The Blood Brain Barrier (BBB) is a highly impermeable and protective cell barrier consisting of tightly sealed endothelial cells. To date, the BBB represents the main hurdle in the development of brain-specific therapeutics, in particular of biologics, such as antisense oligonucleotides (ASOs) and antibodies. Many approaches have been investigated to promote drug transport across the BBB into the parenchyma of the central nervous system (CNS). Notably, receptor mediated transcytosis (RMT) pathways are being exploited, for instance by the engagement of Transferrin Receptors (TfR), Insulin Receptors, and/or Low-density Lipoprotein receptors (LDLR). In our laboratory, we have developed and patented a family of TfR-targeting nanobodies (nTfRs) with efficient BBB-crossing features that have been used for the brain delivery of specific ASOs.

The objectives of this project are: i) to use the existing nTfR technology to target genes involved in the pathobiology of neurological disorders (e.g., BACE1 in Alzheimer’s disease, AD); ii) to improve the BBB-crossing features of the existing nTfR through protein engineering methods.

To accomplish these objectives, ASOs will be designed for selected AD genetic risk factors and synthetized using standard methodology based on solid-phase synthesis and deprotection. Real time qPCR (RT-qPCR) performed on transfected cells overexpressing the targeted gene of interest will be used for in vitro validation of the newly developed ASOs. Molecular cloning techniques and mammalian cell systems (e.g., HEK239T) will be used for the expression of the nTfRs as well as newly engineered nTfR versions. Analytical methods, such as fast protein liquid chromatography (FPLC), will be used for protein purification, while ELISA and surface plasmon resonance (SPR) will be used for binding assessment studies. The best nTfR-ASOs validated in vitro will be injected systemically in wild type mice. RT-qPCR and Western Blot on brain extracts will be used to assess in vivo knock down activity and protein expression, respectively. In situ hybridization will be used to assess ASOs distribution into the brain. The final objective is to test the conjugates in mouse models of AD.

This project is expected to deliver one or more nTfR-ASO conjugates that can effectively cross the BBB and regulate the expression of selected AD genetic risk factors. Therefore, this project will contribute to foster the development of ASOs-based therapeutics that can be used for the treatment of neurodegenerative diseases, such as AD.

The BBB and Neuroinflammation Team of the Institute for Neurophysio-pathology (INP) has a long-standing experience in understanding the role of the BBB during physiological and neuroinflammation conditions. Moreover, with our biotech partner company Vect-Horus we have developed several vector molecules, based on peptides or nanobodies, that facilitate drug transport across the BBB. The INP, is well equipped with all the-state-of-art technological platforms for molecular cloning, cell cultures, fluorescence imaging, molecular interactions assessments, and a new animal facility. The team consists of scientists with all the expertise in BBB physiology and development of brain delivery systems.

The project will be carried by a highly motivated Master student with a degree in biotechnology, biochemistry, molecular sciences, neuroscience, or related fields. The candidate will be required to work in a multicultural and multidisciplinary environment. The candidate is expected to have a strong theoretical knowledge in molecular biology, cell culture (mammalian and bacterial), protein expression and purification, and antibody-antigen biophysical characterization. Computational knowledge for protein modelling and programming skills (e.g., Python) are a plus.