Periodic Reporting for period 3 - DEEPER (DEEP BRAIN PHOTONIC TOOLS FOR CELL-TYPE SPECIFIC TARGETING OF NEURAL DISEASES)
Reporting period: 2024-01-01 to 2025-07-31
DEEPER project (Deep Brain Photonic Tools for Cell-Type Specific Targeting of Neural Diseases) put together technological, neuroscientific and clinical experts with innovative start-ups and leading companies with the aim of developing photonic tools for imaging and manipulating neuronal activity in deep brain regions. The long-term vision of the project is to exploit photonics for meeting medical and research needs in revealing the molecular and cellular dysfunctions underlying the pathogenesis of neurological diseases.
Existing medical tools fail in giving a detailed, high-resolution, dynamic picture of sub-cortical brain structures in behaving animals. It is in this deep region that the pathological behaviour of neurotransmitters such as dopamine, serotonin, and norepinephrine alter the function of brain circuits. This is at the origin of neurological illness with a dramatic social impact such as addiction, chronic pain, Alzheimer’s disease, depression, schizophrenia, and autism spectrum disorder.
The project consortium expects to develop, through neurophotonic techniques, less invasive and more effective treatments for brain neurological conditions. All developed tools will be tested in well-defined and ethically approved animal models of human disease, assessing their suitability for routine use in neuroscience laboratories and their clinical translational potential.
The consortium, thanks to the developed technologies, will push the construction of a value chain that will strengthen Europe’s industrial position in the biophotonics market for microscopy and endoscopy and will accelerate to market the developed tools and project results, via startups and market leaders. This acceleration to commercialization will occur in all the areas covered by the project (genetic tools, pharmaceutical tools, implants, and advanced optical instruments).
Existing medical tools fail in giving a detailed, high-resolution, dynamic picture of sub-cortical brain structures in behaving animals. It is in this deep region that the pathological behaviour of neurotransmitters such as dopamine, serotonin, and norepinephrine alter the function of brain circuits. This is at the origin of neurological illness with a dramatic social impact such as addiction, chronic pain, Alzheimer’s disease, depression, schizophrenia, and autism spectrum disorder.
The project consortium expects to develop, through neurophotonic techniques, less invasive and more effective treatments for brain neurological conditions. All developed tools will be tested in well-defined and ethically approved animal models of human disease, assessing their suitability for routine use in neuroscience laboratories and their clinical translational potential.
The consortium, thanks to the developed technologies, will push the construction of a value chain that will strengthen Europe’s industrial position in the biophotonics market for microscopy and endoscopy and will accelerate to market the developed tools and project results, via startups and market leaders. This acceleration to commercialization will occur in all the areas covered by the project (genetic tools, pharmaceutical tools, implants, and advanced optical instruments).
In the first 18 months of DEEPER, partners focused on developing genetically encoded sensors, optogenetic actuators, and photopharmacological tools for neuromodulation and deep-brain activity recording, enabling real-time tracking of dopamine, norepinephrine, and serotonin. A small library of photoswitchable drugs was initiated for potential light-based treatment of chronic inflammatory pain, while new optogenetic actuators were created for in vivo neurosignaling control. Hardware innovations included tapered fiber optrodes for simultaneous optical and electrical readout, µLED-integrated microelectrodes, and a three-photon laser system improving multimode fiber-based endoscopic imaging with electrophysiology. These technologies supported clinically relevant studies on addiction, chronic pain, Alzheimer’s, and psychiatric disorders using fiber photometry and specialized probes for freely moving animals. Additionally, new methods were developed to investigate developmental miswiring in schizophrenia and autism models, enabling large-scale network analysis in neonatal and juvenile mice.
In the second 18 months of DEEPER, partners advanced molecular tools, implantable probes, and deep-brain imaging systems through collaborations with clinical experts, creating ultrasensitive fluorescent sensors, opsin sequences for synaptic transmission, and photoswitchable compound libraries including red-light activatable drugs. Microfabrication enabled electrode integration on tapered fibers, the development of dual-color silicon probes with micro-LEDs, and flexible substrates combining electrodes and fluidic channels for localized drug delivery and optimal signal recording. Imaging innovations included minimally invasive endoscopes with GRIN lenses, three-photon holographic microscopes, and a two-photon fiber-based micro-endoscope with an extended field of view, complemented by wireless head stages, μLaser arrays, and custom control software (“DeeperScope”). Preliminary system tests validated these technologies, while fiber photometry experiments revealed regional drug responses and dopamine release in freely moving mice. Proof-of-principle studies on Alzheimer’s, schizophrenia, and autism uncovered nonlinear excitatory neuron reorganization during adolescence, opening new paths for understanding neurodevelopmental disorders.
In the final months of DEEPER, the consortium delivered a new class of photonic tools for deep-brain imaging and manipulation, achieving unprecedented penetration depth, resolution, and specific molecular sensors and actuators for neuroscience researches. This platform enabled clinically relevant studies on addiction, chronic pain, Alzheimer’s, and psychiatric disorders, revealing key molecular and cellular dysfunctions. Major achievements include the integration of nonlinear microscopy and innovations in tapered fibers, the transformation of holographic endoscopy for long-term high-resolution studies, the development of ultrasensitive biosensors and pain-control strategies, and the design of dual-color µLED probes and wireless head stages for freely behaving experiments. Additional breakthroughs came from patented µLED implants, identification of critical developmental windows in neonatal and adolescent periods for insights into neurodevelopmental disorders and therapies, and insights into cocaine’s dual action. Complementary advances from different partners introduced cutting-edge optogenetic actuators, photopharmacology, commercialized probe arrays, and depth-selective laser-fiber systems, collectively setting the stage for transformative neuroscience and translational applications.
In the second 18 months of DEEPER, partners advanced molecular tools, implantable probes, and deep-brain imaging systems through collaborations with clinical experts, creating ultrasensitive fluorescent sensors, opsin sequences for synaptic transmission, and photoswitchable compound libraries including red-light activatable drugs. Microfabrication enabled electrode integration on tapered fibers, the development of dual-color silicon probes with micro-LEDs, and flexible substrates combining electrodes and fluidic channels for localized drug delivery and optimal signal recording. Imaging innovations included minimally invasive endoscopes with GRIN lenses, three-photon holographic microscopes, and a two-photon fiber-based micro-endoscope with an extended field of view, complemented by wireless head stages, μLaser arrays, and custom control software (“DeeperScope”). Preliminary system tests validated these technologies, while fiber photometry experiments revealed regional drug responses and dopamine release in freely moving mice. Proof-of-principle studies on Alzheimer’s, schizophrenia, and autism uncovered nonlinear excitatory neuron reorganization during adolescence, opening new paths for understanding neurodevelopmental disorders.
In the final months of DEEPER, the consortium delivered a new class of photonic tools for deep-brain imaging and manipulation, achieving unprecedented penetration depth, resolution, and specific molecular sensors and actuators for neuroscience researches. This platform enabled clinically relevant studies on addiction, chronic pain, Alzheimer’s, and psychiatric disorders, revealing key molecular and cellular dysfunctions. Major achievements include the integration of nonlinear microscopy and innovations in tapered fibers, the transformation of holographic endoscopy for long-term high-resolution studies, the development of ultrasensitive biosensors and pain-control strategies, and the design of dual-color µLED probes and wireless head stages for freely behaving experiments. Additional breakthroughs came from patented µLED implants, identification of critical developmental windows in neonatal and adolescent periods for insights into neurodevelopmental disorders and therapies, and insights into cocaine’s dual action. Complementary advances from different partners introduced cutting-edge optogenetic actuators, photopharmacology, commercialized probe arrays, and depth-selective laser-fiber systems, collectively setting the stage for transformative neuroscience and translational applications.
The advances beyond the state-of-the-art in DEEPER is resulting in a number of unprecedented key exploitable results and market products: (i) HDlight indications are expected to increase ~3 times the dynamic range, to introduce new monitored neurotransmitters and red-shift operative wavelengths; (ii) the consortium will introduce presynaptic silencing tools with 10-times lower activation threshold and light-gated channels with ~5 times higher photocurrent; (iii) DEEPER will make available photoswitchable drugs for controlling dopamine, adrenaline, GABA and serotonin; (iv) depth-selective fiber photometry is providing spatial-resolution in sub-cortical structures of free- moving mice; (v) μLEDs density will be at least doubled together with the introduction of multiple microfluidic channels and multichannel electrophysiology; (vi) we will provide the first systems for correlating functional fluorescence and electrophysiology signals with spatial resolution at both population-wide scale and single cell resolution.
DEEPER is developing the novel concept of multifunctional multi-photon microscopy by combining advanced two-photon and three- photon excitation strategies with multifunctional neural interfaces capable endoscopic detection of functional fluorescence and electro-physiological recordings.
At the same time, the consortium is working to demonstrate the first wireless controlled, integrated, lightweight (< 2g) headstage for depth resolved fiber photometry, optogenetics and electrophysiology recordings in freely moving animals.
These new technologies and protocols are being applied to specific pathologies with the ambition of shedding new light on the mechanism of neurological (addiction, pain, AD) and psychiatric disorders (depression, schizophrenia, autism).
DEEPER is developing the novel concept of multifunctional multi-photon microscopy by combining advanced two-photon and three- photon excitation strategies with multifunctional neural interfaces capable endoscopic detection of functional fluorescence and electro-physiological recordings.
At the same time, the consortium is working to demonstrate the first wireless controlled, integrated, lightweight (< 2g) headstage for depth resolved fiber photometry, optogenetics and electrophysiology recordings in freely moving animals.
These new technologies and protocols are being applied to specific pathologies with the ambition of shedding new light on the mechanism of neurological (addiction, pain, AD) and psychiatric disorders (depression, schizophrenia, autism).