Despite tremendous advances in fundamental neuroscience research, the success rate of neuroscience drug discoveryhas been disappointingly low. Among potential factors contributing to this unfortunate situation is the reality that fairlysimplistic conditions of in vitro assays do not reflect the exquisite complexity of the brain. For example, the brain constantlyreceives signals from the world surrounding us, and controls the functions of many organs after processing of these signals.Yet, typical in vitro assays often do not incorporate any external signals dynamically activating neurons during drugscreening, and instead novel drug candidates for neurological disorders are being routinely evaluated based on their effectson either spontaneous neuronal activity or in end-point assays. Advanced in vitro assays have started incorporating cell stimulation capabilities to address this critical shortcoming.However, existing cell stimulation technologies have some inherent shortcomings that might affect the drug screeningresults in an unpredictable way. For example, optogenetics requires to genetically change neurons by expressing exogenouslight-sensitive ion channels to make neurons fit to be optically stimulated. In addition to being time- and expense-consuming, genetic modifications purely for the sake of the cell stimulation capability are especially not desirable in hIPSC-based neurological disease models, because they might change the model itself. We have recently developed a pioneering optical stimulation platform that do not require genetic modifications ofneurons, and, thus, can provide enormous advantages when hIPSC-based disease models are used in drug screening assays.Our platform is based on our breakthrough graphene-mediated optical stimulation (GraMOS) technology that takesadvantage of unique optoelectronic properties of graphene materials and provides non-invasive modulation of the cellactivity via an external light-controlled electrical field near the graphene-neuron biointerface. Here we are proposing to integrate our GraMOS technology into drug screening assays for neuroscience drugdiscovery. We will use 2-D and 3-D hiPSC-based models generated from patients carrying mutations, known to alter theneuronal activity such as the lack of MECP2 gene. For classical 2-D models, we will fabricate graphene-based substratesto enable optical activation of neurons via the bottoms of specialized cell culture plates. We will proceed with thedevelopment of all-optical assays that combine GraMOS-enable optical stimulation with recordings of the neuronal activityusing fluorescent indicators. To evaluate the synaptic connectivity of neuronal networks, we will develop patterned graphenesubstrates to be able to selective stimulate only a subset of neurons interfacing with G-substrates. In 3-D models, such asneuronal spheroids and brain cortical organoids, we will evaluate the functional neuronal activity by either using all-opticalGraMOS-enabled assays or combining GraMOS with MEA-based recording of extracellular electric activity. GraMOS-enabled assays will be validated using benchmark compounds affecting synaptic transmission and neuronal excitability.The proposed non-genetic optical stimulation technology is expected to dramatically enhance translational neurosciencestudies and support the discovery of neurological drugs with novel mechanism of actions.
Public Health Relevance Statement: PUBLIC HEALTH RELEVANCE STATEMENT
The quest for enhanced understanding the brain function and predictive drug discovery for neurological
disorders requires advanced technological tools that can probe and modulate the activity of neural networks
without interfering with their genetic identity. To address this need and make neuroscience drug discovery
process more predictive, we are proposing to develop a novel nanotechnology-based platform for optical
stimulation of genetically intact neurons and incorporate it into the next generation drug screening assays. The
proposed study will result in tremendous technological benefits to fundamental and translational studies that are
probing the synaptic connectivity of neuronal networks in health and disease and striving to discover new drugs
for a wide range of neurological disorders.
Project Terms: Adoption ; Affect ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Biotechnology ; Biotech ; Brain ; Brain Nervous System ; Encephalon ; Calcium ; Cell Communication ; Cell Interaction ; Cell-to-Cell Interaction ; Cell Culture Techniques ; cell culture ; Cells ; Cell Body ; Communication ; Data Analyses ; Data Analysis ; data interpretation ; Disease ; Disorder ; Pharmaceutical Preparations ; Drugs ; Medication ; Pharmaceutic Preparations ; drug/agent ; Electrophysiology (science) ; Electrophysiology ; Neurophysiology / Electrophysiology ; electrophysiological ; Environment ; Exhibits ; Genes ; Genetic Identity ; Health ; Human ; Modern Man ; Hybrids ; In Vitro ; Ion Channel ; Ionic Channels ; Membrane Channels ; Lead ; Pb element ; heavy metal Pb ; heavy metal lead ; Light ; Photoradiation ; Neurological Models ; Neurologic Models ; Mutation ; Genetic Alteration ; Genetic Change ; Genetic defect ; genome mutation ; nervous system disorder ; Nervous System Diseases ; Neurologic Disorders ; Neurological Disorders ; neurological disease ; Synaptic Transmission ; Neural Transmission ; Neurons ; Nerve Cells ; Nerve Unit ; Neural Cell ; Neurocyte ; neuronal ; Neurosciences ; Optics ; optical ; Organoids ; Patients ; Pharmacology ; Physiology ; Signal Transduction ; Cell Communication and Signaling ; Cell Signaling ; Intracellular Communication and Signaling ; Signal Transduction Systems ; Signaling ; biological signal transduction ; Synapses ; Synaptic ; synapse ; Technology ; Time ; G-substrate ; cerebellum protein substrate for cGMP dependent protein kinase ; protein G ; Mediating ; base ; Organ ; culture plates ; Physiological ; Physiologic ; Neurologic ; Neurological ; Ensure ; Chemicals ; Evaluation ; Disease Progression ; Spottings ; Genetic ; Morphology ; tool ; electric field ; Pattern ; System ; 3-D ; 3D ; three dimensional ; 3-Dimensional ; extracellular ; Best Practice Analysis ; Benchmarking ; empowered ; biocompatibility ; biomaterial compatibility ; calcium indicator ; experience ; light intensity ; success ; voltage ; Structure ; advanced illness ; advanced disease ; novel ; novel technologies ; new technology ; disorder model ; Disease model ; Modeling ; Property ; drug development ; nano tech ; nano technology ; nano-technological ; nanotech ; nanotechnological ; Nanotechnology ; Endpoint Assays ; End Point Assay ; drug discovery ; Pharmaceutical Agent ; Pharmaceuticals ; Pharmacological Substance ; Pharmacologic Substance ; Thickness ; Thick ; Address ; Multiparametric Analysis ; in vitro Assay ; in vivo ; Neurosciences Research ; Optical Methods ; Screening Result ; Monitor ; Process ; Modification ; Development ; developmental ; Image ; imaging ; three-dimensional modeling ; 3-D modeling ; 3D modeling ; next generation ; neuronal excitability ; nanoscience ; nano science ; Outcome ; Consumption ; innovation ; innovate ; innovative ; genetically modified cells ; genetically engineered cells ; translational study ; novel therapeutics ; new drug treatments ; new drugs ; new therapeutics ; new therapy ; next generation therapeutics ; novel drug treatments ; novel drugs ; novel therapy ; public health relevance ; non-genetic ; nongenetic ; translational neuroscience ; drug candidate ; optogenetics ; screening ; signal processing ; axion ; graphene ; experimental study ; experiment ; experimental research ; imaging study ; Drug Screening ; neural network ; side effect ; disease-in-a-dish ;
