Periodic Reporting for period 3 - NEUREKA (A smart, hybrid neural-computo device for drug discovery)
Okres sprawozdawczy: 2022-12-01 do 2024-05-31
The NEUREKA project aims to transform drug discovery for neurological diseases, which currently represent a major economic and societal burden. With over one billion people affected globally and treatment costs reaching €1.4 trillion annually in the EU and US, the need for effective therapies is urgent. Despite significant investments from pharmaceutical companies, drugs targeting brain disorders have a 45% lower success rate compared to other drug classes, widening the gap between demand and efficacy.
To combat this issue, NEUREKA focuses on strengthening the functional screening of compounds through in vitro cellular assays. While these assays are valuable for studying brain circuits, they often fail to replicate crucial cell-to-cell interactions found in native tissues. This shortcoming hinders accurate modeling of brain pathology, as cultured neurons typically lack essential in-brain synaptic inputs. Current technologies provide limited control over network excitability and plasticity, focusing mainly on single-cell observations. This makes it challenging to assess how molecular deficits and treatments impact neuronal circuits.
We propose an innovative hybrid technology that combines nanoelectrodes with computational models of neuronal circuits. This will allow precise manipulation and measurement of activity in cultured networks related to Alzheimer’s disease. For the first time, a biophysical model will drive dendritic stimulation, replicating both pathological connectivity and molecular features of the disease. By enhancing drug discovery success rates, our approach aims to reduce costs and minimize animal use in preclinical trials.
To combat this issue, NEUREKA focuses on strengthening the functional screening of compounds through in vitro cellular assays. While these assays are valuable for studying brain circuits, they often fail to replicate crucial cell-to-cell interactions found in native tissues. This shortcoming hinders accurate modeling of brain pathology, as cultured neurons typically lack essential in-brain synaptic inputs. Current technologies provide limited control over network excitability and plasticity, focusing mainly on single-cell observations. This makes it challenging to assess how molecular deficits and treatments impact neuronal circuits.
We propose an innovative hybrid technology that combines nanoelectrodes with computational models of neuronal circuits. This will allow precise manipulation and measurement of activity in cultured networks related to Alzheimer’s disease. For the first time, a biophysical model will drive dendritic stimulation, replicating both pathological connectivity and molecular features of the disease. By enhancing drug discovery success rates, our approach aims to reduce costs and minimize animal use in preclinical trials.
The project was completed on May 31st, 2024. All scientific, management, dissemination, communication and exploitation activities were successfully completed. A summary of the main results is provided below:
1) New Nanowire Array technologies: we developed a new in-vitro platform for neuron-on-a-chip interfacing through nanowires. We performed extensive studies of various materials of the PtSi layer, including the surface oxidation for a proper integration of the coating layers. We also fabricated a 60-channel NWA and established the procedure for electrical characterization of such a system in recording (impedance) and stimulation. We developed a low temperature NWA process compatible with an integration on CMOS circuit and performed a first line of nano-electrode arrays that were integrated on CMOS.
2) New biophysical model of Alzheimer’s disease: we developed the AlzModel software, a detailed, multiscale, neuronal network model that reproduces key deficits of Alzheimer’s disease such as impaired synaptic plasticity/rigidity, decreased NMDA currents, dendritic atrophy etc.
3) The Synaptor: we developed a computational model of the Synaptor, which served as the interface between the nanoelectrodes and the neurons that were cultured on the surface of the chip. The Synaptor was developed and validated with electrophysiological data from rat hippocampus neuronal cultures on nanowire array prototypes and on the high-density planar multi-electrode array provided by our industrial partner.
4) The integrated NEUREKA system: we established a controllable hybrid system, whereby a hardware chip device (multi- or nanowire electrode array) is interfaced with the AlzModel and used to control the activity of cultured neurons on the surface of the array. Software pipelines were also developed to ensure a seamless exchange of data between the AlzModel and the chip in a bidirectional manner. The integrated NEUREKA system was implemented using two different platforms: the 60-channel nanowire array and the high density multielectrode array MaxOne.
5) Testing of the NEUREKA platform: we tested the ability of the AlzModel to successfully drive disease-related states on both chip platforms, using rat and hiPSC cultures. To do so, we developed a software package for automatic detection and functional characterization of subcellular axonal physiological signals, across hundreds of neurons within a neuronal network. We also established protocols for stable and reproducible culturing of human iPSC-derived glutamatergic neurons and human astrocytes.
4) Drug testing pilot: we tested the ability of the NEUREKA platform to serve as a drug screening device using both rat and hiPSC cultures. Abeta homogenate was used to induce the Alzheimer’s phenotype on cultured neurons and the effectiveness of known drugs (memantine and valproic acid) on the activity and connectivity properties of cultured neurons, when driven by the AlzModel, was assessed. Our results suggest that NEUREKA has the potential to serve as a reliable drug screening device.
Exploitation activities:
The nanoelectrode technology, implemented on various substrates (including CMOS chips), has been granted a patent in France (CNRS-LAAS), with several international extensions still pending. CNRS is actively promoting the transfer of this technology to an industrial environment for commercialization and wider application.
A second patent for the AlzModel as an integrated system for drug screening is being drafted (FORTH-UNIPD). Efforts are made to secure funding to further develop both of the technologies into marketable products (e.g. through EIC Transition applications).
1) New Nanowire Array technologies: we developed a new in-vitro platform for neuron-on-a-chip interfacing through nanowires. We performed extensive studies of various materials of the PtSi layer, including the surface oxidation for a proper integration of the coating layers. We also fabricated a 60-channel NWA and established the procedure for electrical characterization of such a system in recording (impedance) and stimulation. We developed a low temperature NWA process compatible with an integration on CMOS circuit and performed a first line of nano-electrode arrays that were integrated on CMOS.
2) New biophysical model of Alzheimer’s disease: we developed the AlzModel software, a detailed, multiscale, neuronal network model that reproduces key deficits of Alzheimer’s disease such as impaired synaptic plasticity/rigidity, decreased NMDA currents, dendritic atrophy etc.
3) The Synaptor: we developed a computational model of the Synaptor, which served as the interface between the nanoelectrodes and the neurons that were cultured on the surface of the chip. The Synaptor was developed and validated with electrophysiological data from rat hippocampus neuronal cultures on nanowire array prototypes and on the high-density planar multi-electrode array provided by our industrial partner.
4) The integrated NEUREKA system: we established a controllable hybrid system, whereby a hardware chip device (multi- or nanowire electrode array) is interfaced with the AlzModel and used to control the activity of cultured neurons on the surface of the array. Software pipelines were also developed to ensure a seamless exchange of data between the AlzModel and the chip in a bidirectional manner. The integrated NEUREKA system was implemented using two different platforms: the 60-channel nanowire array and the high density multielectrode array MaxOne.
5) Testing of the NEUREKA platform: we tested the ability of the AlzModel to successfully drive disease-related states on both chip platforms, using rat and hiPSC cultures. To do so, we developed a software package for automatic detection and functional characterization of subcellular axonal physiological signals, across hundreds of neurons within a neuronal network. We also established protocols for stable and reproducible culturing of human iPSC-derived glutamatergic neurons and human astrocytes.
4) Drug testing pilot: we tested the ability of the NEUREKA platform to serve as a drug screening device using both rat and hiPSC cultures. Abeta homogenate was used to induce the Alzheimer’s phenotype on cultured neurons and the effectiveness of known drugs (memantine and valproic acid) on the activity and connectivity properties of cultured neurons, when driven by the AlzModel, was assessed. Our results suggest that NEUREKA has the potential to serve as a reliable drug screening device.
Exploitation activities:
The nanoelectrode technology, implemented on various substrates (including CMOS chips), has been granted a patent in France (CNRS-LAAS), with several international extensions still pending. CNRS is actively promoting the transfer of this technology to an industrial environment for commercialization and wider application.
A second patent for the AlzModel as an integrated system for drug screening is being drafted (FORTH-UNIPD). Efforts are made to secure funding to further develop both of the technologies into marketable products (e.g. through EIC Transition applications).
The NEUREKA project aims to revolutionize drug discovery through a groundbreaking hybrid platform that uniquely drives, modulates, and reads neuronal connectivity and activity of cultured neurons under realistic disease conditions. This innovative approach is expected to enhance the discovery of effective treatments for neurodegenerative diseases, which pose a significant economic and societal burden in Europe. As the aging population increases, the prevalence of these diseases will rise, making solutions even more critical.
NEUREKA has generated new knowledge and technological advancements across various fields, including the development of new materials for nanowire conductivity and novel devices, such as nanowire arrays, that enable unprecedented resolution in stimulating neurons. The project also introduced powerful software for real-time processing and control of electrical signals in model neurons, alongside new biophysical models of Alzheimer’s disease that realistically simulate pathology in large-scale networks.
Key advancements include:
A) Integration of the AlzModel with nano- and multielectrode arrays.
B) Demonstration of stimulating both nano- and multi-electrodes via the AlzModel.
C) Ability of the AlzModel to replicate realistic Alzheimer’s activity states in cultured neurons.
D) Validation of the platform for drug testing, yielding reliable activity and connectivity readouts.
Additionally, NEUREKA has positively impacted the training of 25 young researchers and is poised to create significant market opportunities. It addresses the need for high-throughput devices to measure intracellular signals and accurate methods to screen numerous human samples generated by iPSC technology.
NEUREKA has generated new knowledge and technological advancements across various fields, including the development of new materials for nanowire conductivity and novel devices, such as nanowire arrays, that enable unprecedented resolution in stimulating neurons. The project also introduced powerful software for real-time processing and control of electrical signals in model neurons, alongside new biophysical models of Alzheimer’s disease that realistically simulate pathology in large-scale networks.
Key advancements include:
A) Integration of the AlzModel with nano- and multielectrode arrays.
B) Demonstration of stimulating both nano- and multi-electrodes via the AlzModel.
C) Ability of the AlzModel to replicate realistic Alzheimer’s activity states in cultured neurons.
D) Validation of the platform for drug testing, yielding reliable activity and connectivity readouts.
Additionally, NEUREKA has positively impacted the training of 25 young researchers and is poised to create significant market opportunities. It addresses the need for high-throughput devices to measure intracellular signals and accurate methods to screen numerous human samples generated by iPSC technology.