Protein misfolding diseases (PMDs) are a large group of human disorders caused by the misfolding of specific proteins. They include conditions with high socio-economic impact, such as Alzheimer’s disease, cystic fibrosis, obesity and type 2 diabetes, and the majority of them remain incurable. Among the >70 PMDs, about 1/3 are caused by misfolding-prone membrane proteins (MisMPs). Despite their significance, and mainly due to the difficulties associated with MisMP overexpression, isolation and characterization, the PMD scientific community has largely overlooked MisMP-associated PMDs, thus limiting opportunities for drug discovery. In ProMisMe, we will develop engineered bacteria and yeast, which can function as broadly applicable discovery platforms for compounds that rescue MisMP misfolding. These compounds will be selected from libraries of drug-like molecules biosynthesized in these microbes using a technology that allows the facile production of tens of millions to tens of billions of different test molecules. These libraries will then be screened in the same microbial cells that produce them and the rare molecules that rescue MisMP misfolding effectively will be selected by ultrahigh-throughput screening. The effect of the selected molecules on MisMP folding will then evaluated by biochemical and biophysical methods, while their ability to reverse MisMP-induced pathogenicity will be tested in appropriate mammalian cell assays. The molecules that rescue the misfolding and associated pathogenicity of the target MisMPs will become therapeutic candidates against the corresponding diseases. This procedure will be applied for different MisMPs to identify potential therapeutics for two serious PMDs: Usher Syndrome III and Charcot-Marie-Tooth disease. Successful realization of ProMisMe will provide invaluable therapeutic leads against major diseases and a more widely applicable framework for drug discovery against diseases caused by membrane protein misfolding.

The Biomedical Sciences Research Center “Alexander Fleming” (FLEMING) is the top-ranked research center in the Life Sciences in Greece. FLEMING has gained international recognition for its pioneering research towards understanding the molecular basis of human diseases, such as autoimmune disorders, cancer, neurodegeneration and osteoporosis, and the development of new approaches for their diagnosis and treatment. The Centre’s major goal for the coming years is to support excellent research with a focus on innovation. In this direction, FLEMING has recently established the new Institute of Bio-Innovation with a mission to advance basic research discoveries towards innovation in the areas of biotechnology and drug discovery and convert them to tangible outcomes for the economy and the society. Furthermore, FLEMING has planned the establishment of the national R&I infrastructure “Biotechnopolis”, the first integrated Biotechnology hub in Greece and the wider region, to foster the development of innovative biomedical, biotechnological and pharmaceutical products and technologies through the creation of a fertile environment offering access to excellent research, high-end infrastructures and a culture of entrepreneurship close to academia. The goal of Boost4Bio is to attract and maintain an exceptional researcher and innovator in Biomolecular Engineering & Synthetic Biology to establish a new excellent team at FLEMING, promote the integration of basic and applied research, boost bio-innovation and bio-entrepreneurship at the Center, and lead the establishment of “Biotechnopolis”. Boost4Bio will have a major impact on the promotion of excellence in biomedical research, training and innovation in Greece, will support the proper establishment of a new and unique Excellence Unit in Greece and the wider region, and will contribute to the creation of new career prospects and the prevention of the brain drain phenomenon, which is particularly acute in this area of the Union.

Lung cancer is the leading cause of cancer death. Immunotherapy improved survival rates, but efficacy is limited to selected patients. We recently discovered universal antigen presenting fibroblasts (apFibros) across human and murine lung tumors and showed that they directly stimulate cancer-specific CD4 T cells, creating immunological hot spots that support immune rejection. These studies achieved a breakthrough on the role of in situ cancer antigen presentation and proposed a novel model whereby tumors can sustain T cells independently of lymph nodes. Preliminary data suggest that lung apFibros help overcome resistance to checkpoint inhibitors. For their immunotherapeutic exploitation of apFibros two bottlenecks must be overcome: low numbers and incomplete understanding of their configurations. We will integrate computational and laboratory experiments and work in parallel in human and mouse models to generate perturbation datasets across single-cell/cell systems, transcriptomics/epigenomics, spatial/temporal levels, and dissect the molecular landscape that regulates fibroblast states. Our ultimate goal is to unravel perturbations that can diverge cancer-associated fibroblasts to antigen presenting states. The following questions are at the core of our proposal i) how do diverse fibroblast states emerge and evolve? ii) which gene regulatory networks drive specificity of these states? ii) which are the functional modules that are driven by apFibros and how are they mechanistically explained? iv) how can we transdifferentiate existing fibroblasts to acquire antigen presenting states? v) how can fibroblast reprogramming help overcome immunotherapy resistance? The proposed research should help advance mechanistic concepts in what we term the “adaptive immune mesenchyme”, decode the complexity of peripheral antigen presentation in tumors and beyond and promote targeting of the stroma for immunotherapy.