ENGINEERING PROBIOTICS FOR THE SAFE DELIVERY OF THERAPEUTIC PROTEINS IN THE TREATMENT OF RARE NEURODEVELOPMENTAL DISORDERS

Most existing strategies for the treatment of diseases focus on the delivery of naked molecules with a therapeutic activity, from chemically synthesized molecules to recombinant proteins produced in diverse platforms such as bacteria, yeast, insect cells, mammalian cells, and viruses. However, in many cases these treatments require the use of invasive administration methods such as intravenous, subcutaneous, or in situ injection of the molecule of interest in order to reach the targeted region. Given the necessity to develop effective and safe strategies for the delivery of therapeutic molecules in the treatment of neurological disorders, Lactic Acid Bacteria (LAB) may present an appealing option. LAB have been used for centuries in food production and preservation, and as probiotic agents, and have a long history of safe exploitation by humans. The innovative idea of this proposal, therefore, is to use LAB, which are capable of reaching and colonizing the intestine, to deliver and produce secretable TATk-fused proteins that, due to the transduction property of the TATk peptide, are able to pass the blood-brain barrier and reach the brain. Since they are live organisms, LAB would be able to autonomously multiply, produce, and deliver the therapeutic protein. By exploiting a mouse model of CDD, the Cdkl5 KO mouse, we will study the effect of in vivo LAB-mediated TATk-CDKL5 production. The successful outcome of the proposed project will establish the proof-of-principle of LAB as a safe delivery platform of recombinant CDKL5 protein for the treatment of CDD.