ND Biosciences’ Protein Engineering Technologies Allow Reproducing the Complexity and Diversity of Relevant Targets for Drug and Biomarker Discovery
ND Biosciences’ powerful proprietary technologies enable for the first time reproducing, at both the structural and biochemical levels, the complexity and diversity of monomeric and pathological forms of the disease-causing proteins found in the brain and biological fluids of patients suffering from different neurodegenerative diseases (NDs). Thus, ensuring the development of disease-relevant therapies and diagnostics.
Chemical Synthesis of Proteins
Although post-translational modifications (PTMs) such as phosphorylation, ubiquitination, nitration, and truncation have been implicated in modulating the process of protein aggregation and related toxicity, most screening assays and drug development programs today are still based on using unmodified proteins or platforms that do not capture the true biochemical properties of related proteins. This was mostly due to the lack of enabling technologies and platforms.
Based on 15 years of R&D at the EPFL, ND Biosciences is now capable of introducing site-specific PTMs at any position along the sequence of proteins implicated in NDs. This allows the recapitulation of both the biochemical and structural properties of these proteins, and the generation of libraries of proteins bearing either single or multiple PTMs, but also in the context of relevant protein conformations. Importantly, these technologies have been successfully applied for the modification and large scale production of libraries of several ND-causing proteins including α-Synuclein, Tau, Huntingtin and TDP43, which are key players in the pathogenesis PD, AD, Huntington’s disease and Amyotrophic lateral sclerosis, respectively. These libraries are utilized by ND Biosciences for the development, identification and profiling of novel binders with therapeutic potential, and for the development of sensitive and accurate diagnostic assays and imaging agents that capture the diversity of pathologically relevant species in different diseases.
Reproducing the biochemical and structural diversity of protein aggregates
At the structural level, ND Biosciences’ technologies allow reproducing the variety of conformations of misfolding proteins, including unfolded monomers, vesicle bound structures, disease-relevant oligomers with distinct conformations and morphologies, and different conformational strains of fibrillized aggregates. These advances ensure that our therapies and diagnostics will efficiently target the diversity of the disease-causing proteins and their pathologies.
One of ND Biosciences’ powerful protein engineering approaches, pioneered at the EPFL, allows generating pure protein fibrils that mimic the structural and conformational properties of fibrils purified from brains of patients suffering from Alzheimer’s disease (AD). In previous technologies utilized for drug discovery, the structure, morphology and biochemical features of generated fibrils do not reproduce that of protein aggregates typically observed in human pathology, therefore explaining the failure of these discovery programs targeting irrelevant preparations. ND Biosciences is the only company with an exclusive license to patent-pending enabling technology that allows reproducing the correct conformation of pathological Tau, one of the two main proteins implicated in the pathogenesis of AD. Importantly, this technology allows the production of relevant targets at a scale that is amenable for drug and biomarker discovery.
At the structural level, ND Biosciences’ technologies allow reproducing the variety of conformations of misfolding proteins, including unfolded monomers, vesicle bound structures, different oligomeric configurations and morphologies, and different conformational strains of fibrillized aggregates.
ND Biosciences’ Integrated Drug Discovery Platforms Allow Dissecting the Mode of Action of Candidate Therapeutics
ND Biosciences’ drug discovery platforms include an integrated suite of models that reproduce not only the key pathological signatures, but also relevant processes underlying protein aggregation in human brain pathology. These models include: i) cell-free systems allowing real-time assessment of protein aggregation kinetics in vitro, ii) complex cellular and neuronal models that reproduce different stages and key processes underlying protein aggregation, toxicity and Lewy body formation in neurons, and (iii) different animal models of protein aggregation and related neurotoxicity and behavioral deficits. These integrated platforms enable comprehensive assessment of the anti-aggregation properties of potential therapeutic molecules, as well as determining their mode of action.
As ND Biosciences’ co-founders recently discuss in an elaborate review published in Nature Reviews Neurosciences, the great majority of neuronal models currently used in Parkinson’s disease (PD) research do not reproduce the structural features or key processes underlying protein aggregation in human brains. This could be a key factor as to why most therapeutic candidates that show promising results in existing models fail in the clinic. Based on 10 years of R&D at the EPFL, ND Biosciences drug discovery platforms comprise a unique portfolio of neuronal models that reproduce distinct stages of protein pathology, and allow interrogation of therapeutic effects at early and late stages of diseases. These models were characterized at unprecedented detail using a combination of biochemical, imaging, and proteomic techniques, and include neuronal models that mimic: i) early stages of de novo inclusion formation in Parkinson’s disease on the basis of expressing α-Syn in a specific strain of mice (Fares et al., 2016), ii) extracellular protein aggregation and mediated toxicity upon treatment with specific mixtures of pathologic α-Syn preparations (Mahul-Mellier et al., 2015), iii) intermediate stages of α-Syn protein uptake and seeding of endogenous α-Syn aggregation following treatment with preformed α-Syn fibrils (Mahul-Mellier et al., 2019), and iv) late stages of α-Syn aggregation into Lewy body (LB) inclusions that is concomitant with neurotoxic effects, phenomena that were previously unattainable in any of the previous models utilized for drug discovery (Mahul-Mellier et al., 2020). Utilizing these models, ND Biosciences is now able to reproduce different pathological stages and processes in living neurons, and to develop small molecules and biologics that specifically target these aberrant mechanisms.