PARTICIPATION IN HEALTH PARTNERING DAY 2017

The overall scientific objective of this study is to develop a cell-targeted gene therapy approach using clinically translatable viral vectors. This work will facilitate the treatment of common inherited demyelinating neuropathies caused by mutations in genes expressed by myelinating Schwann cells. Adeno-associated viral (AAV) vector serotypes including AAV9, AAVrh10, AAV.PHP.S, AAV2/rh10 will be evaluated for their efficiency to infect cultured human Schwann cells. In addition, minimal myelin-specific promoters such as the minimal Mpz and the Aatk will be cloned and their efficacy to drive cell-targeted expression will be evaluated in myelinating Schwann cell co-cultures. Their Schwann cell-specific gene expression will be confirmed following in vivo delivery. AAV serotypes with highest Schwann cell tropism and minimal promoters with Schwann cell-specific expression will be used to produce therapeutic AAV vectors, which will be tested in peripheral neuropathy models including CMT1A and CMT1X models. The GJB1 gene mutated in CMT1X neuropathy will be cloned under the minimal promoter and will be delivered to Cx32 knockout mice to analyze expression and localization of Cx32. PMP22-targeting shRNAs will be also be cloned under Schwann cell specific promoters and delivered to the C61 mouse model of CMT1A overexpressing human PMP22. Thus, we will use a gene addition approach for CMT1X and gene silencing for CMT1A. Finally, we will proceed with a treatment trial in both CMT models. The level of phenotypic rescue will be evaluated by motor behavioral, electrophysiological and morphological methods 4 months after treatment compared to mock-treated littermates, in order to provide a proof of principle for clinical translation.

According to the Alzheimer Disease (AD) amyloid hypothesis, the deposition of prefibrillar and fibrillar alpha-beta peptide sets off pathogenic cascades of neuroinflammation and neurodegeneration that lead to synaptic and neuronal loss and cognitive decline. The complement cascade has been found to modulate the disease even though the mechanism of action is highly controversial. Animal experiments in the AD mouse model show that inhibition of the complement component C5a (a terminal product of complement activation) reduces amyloid and alleviates the pathology. Macrophages and neutrophils carry C5a receptors. In the mouse model for ATTR V30M neuropathy, C5a receptor inhibition exacerbates amyloid deposition while administration of the full C5a agonist significantly reduces amyloid deposition. Yet, inhibition of the C5a receptor in the AD mouse model resulted in the decrease of amyloid deposition. We believe the difference may lie in the fact that in TTR amyloidosis the amyloidogenic peptides are imported from the circulation (produced in the liver), while in AD the alpha-beta peptides are locally produced so that prolonged C5a stimulation may actually increase production of alpha-beta due to the increase in inflammation. C5a receptor agonists have not been fully tested in AD and we hypothesize, based on preliminary data, that in fact they will have a beneficial effect by clearing amyloid. Thus, a modified C5a receptor agonist along with a C5a receptor antagonist will be intermittently administered to AD mice by mouth. Following the completion of the treatment protocols, Thioflavin S, immunofluorescence, ELISA, immunoblotting, gene expression assays and cognitive testing will be carried out to evaluate the effects of the aforementioned therapeutic molecules on the pathophysiology of the disease.

Haemoglobinopathies are the most common monogenic diseases, with millions of carriers and patients worldwide. In the particular instance of Cyprus, 12% - 15% of the population are carriers of beta-thalassaemia, making it one of the commonest single-gene disorders on the island. Online resources for haemoglobinopathies are largely divided into specialised sites catering for patients, researchers and clinicians separately. However, the severity, ubiquity and surprising genetic complexity of the haemoglobinopathies call for an integrated website as a free and comprehensive repository and tool for patients, scientists and health professionals alike. The ITHANET community portal (http://www.ithanet.eu) is an expanding resource for clinicians and researchers dealing with haemoglobinopathies. It integrates information on news, events, publications, clinical trials and haemoglobinopathy-related organisations and experts, wiki-based content of protocols, clinical guidelines and educational articles and, most importantly, databases of variations, epidemiology and diagnostic and clinical data. The proposed project aims to expand the functionality and content of the ITHANET portal and strengthen its international role as a reference point for haemoglobinopathy-related research, treatment and diagnosis and as a daily scientific resource for patients, carriers, and all those interested in haemoglobinopathies. This will be achieved by (1) classifying and interpreting haemoglobinopathy-related variants based on internationally established standards and guidelines, (2) performing worldwide epidemiological studies, with a focus on micromapping and the impact of migration, and (3) developing the first haemoglobinopathy-specific genotype – phenotype database.

Each year, millions are affected by breast cancer worldwide. While genes like BRCA1 and BRCA2 are responsible for more than 15% of breast cancers, the impact of numerous genetic variants within these high-risk genes is not fully understood. These Variants of Uncertain Significance (VUS) make the clinical management challenging for carriers (identified in 5-10% of the genetic tests performed in BRCA1 and BRCA2; higher for less studied genes). The primary objective of this proposal is to enhance the classification rate of VUS in breast cancer predisposition genes. We plan to use the world's most extensive multi-ancestry case-control dataset and develop cutting-edge software that is going to be broadly and freely applied.Understanding VUS will lead to a better management of carriers and their relatives, avoiding over or under surveillance. Genetic screening allows people to gain a deeper insight into their predisposition to cancer, possibly preventing its onset and increasing their targeted therapy options. This advantage is limited to those who identify their genetic susceptibility, and the identification of a VUS hampers it. This proposal is timely and aligns well with the EU mission on cancer, focusing on understanding genetic susceptibility in breast cancer, identifying relevant pathogenic variants allowing for prevention and early detection strategies, and ultimately improving the lives of patient carriers and their relatives.The scientific component of the project is divided into three work packages (WPs), each with specific objectives ranging from data collection to the development of state-of-the-art methodologies. Successful execution of this proposal could significantly contribute to personalised cancer prevention and treatment strategies and can be adapted for other diseases with a genetic component.