WO2021213758A1 - System comprising genetically modified cells and a removable device - Google Patents

Abstract

The present invention relates to an advantageous system comprising genetically modified cells as well as a removable device, wherein the genetically modified cells are cells of the dermis, cells of the epidermis and/or LTMRs and said genetically modified cells are preferably activated by a light signal emitted by the removable device.

Description

SYSTEM COMPRISING GENETICALLY MODIFIED CELLS AND A REMOVABLE DEVICE

The present invention relates to advantageous system comprising genetically modified cells as well as a removable device, wherein the genetically modified cells are cells of the dermis, cells of the epidermis and/or LTMRs and said genetically modified cells are preferably activated by a light signal emitted by the removable device. This system can be used for sensory enhancement or for creating new sensory means in a subject, in particular a means allowing the perception for example of physical, chemical and/or biological signals which are not perceived by a sense of the subject.

BACKGROUND

Human brain contains about 86 billion neurons and about 100 trillion synaptic connections forming networks with a set of nodes and connections (i.e., a complex set of relationships or circuits).

Neurons from our peripheral nervous system receive and convey signals (i.e., information), and neurons from our central nervous system process (i.e., neural coding) these signals, the processed signals being at the origin of our perception of the world. Our brain is trained to perceive the world via the stimulation of our five natural senses, namely touch, sight, hearing (and balance), smell and taste. In each neural circuit, neurons convey and process information (i.e., neural coding) using electrical signals (also identified as “electrical impulses”, “electrical spikes” or “action potentials”), whatever the receptors responsible for transmitting the information, i.e., the mechanoreceptors, thermoreceptors or pain receptors involved in touch, the photoreceptors involved in sight, the mechanoreceptors involved in hearing or balance, and the chemoreceptors involved in smell or taste.

This processed information (i.e., neural coding) is dynamically represented by patterns of action potentials generated by neurons in relevant brain regions corresponding to moment-to-moment perceptions, memories, creative thoughts and behaviors.

A first type of proposed neural coding is referred to as the “rate code” model. This model considers that information about the stimulus is encoded by the firing rate of the neurons. In practice, the rate is measured by averaging the number of spikes per second, or a defined (often smaller) time bin before and after stimulus presentation and typically over multiple stimulus trials. This averaging procedure inherently assumes that spike variability reflects noises, and most, if not all, information is conveyed by spike numbers.

A second proposed type of neural coding model is referred to as the “temporal code” model. This model utilizes timing information of spike’s discharges to identify the stimulus.

A third proposed type of neural coding model is the “neural self-information” model which postulates that neuronal variability carries itself information. In other words, under this “self information code model”, any given Inter-Spike-Interval (ISI) is self-tagged with a discrete amount of information [Meng Li el al. Neural Code — Neural Self-information Theory on How Cell-Assembly Code Rises from Spike Time and Neuronal Variability. Frontiers in Cellular Neuroscience, 2017; Volume 11; Article 236]

Understanding/decoding the neural coding (i.e., how information is processed) is still an area of investigation.

Brain Machine Interface (BMI) (also identified as Brain Computer Interface (BCI)) can be defined as a direct communication pathway, through any artificial means, that allows the brain to exchange information directly with an external device. BMI may help understanding how the brain encodes sensory information from the outside world into an internal language, how it integrates external and internal information to produce cognitive/emotional representations and how it generates and executes motor programs [Karen A. Moxon el al. Brain-Machine Interfaces beyond Neuroprosthetics. Neuron 86, April 8, 2015; 54-67] BMI is seen as a promising means to not only understand but also to achieve neural coding.

In this context, Neuralink has developed ultra-thin multi-electrode polymer probes which are to be inserted in a mammalian brain, offering the possibility of recording neural activity in real time to decode neural information and also the possibility of modulating neural activity to encode new neural information.

However, direct access to the brain may represent a risk to the subject. In addition, with more than about 86 billion of neurons in a human brain, the possibility to get access to even a fraction of neurons remains low.

Moving away from the central nervous system, the peripheral nervous system appears as an interesting alternative for neural coding and peripheral nerve procedures are associated with less risk to the subject.

Sensory restoration devices have so far used the peripheral nervous system to transmit typically sound information (cochlear implants) or image information (optical implants) via implanted (micro)electrodes arrays that send signals (i.e., electrical signals) directly to the appropriate nerve, auditory nerve or optical nerve respectively.

Sensory substitution devices have used the peripheral nervous system too, to stimulate the receptors of intact natural senses. Eyes have the highest capacity for conveying information followed by the ear and the skin. The ear has the highest temporal resolution. So far, it has been possible to “see through the ear or skin” or to “hear through the eyes or skin” [Meike Scheller el al. Chapter 15, Perception and Interactive Technology. Stevens’ Handbook of Experimental Psychology and Cognitive Neuroscience, Fourth Edition, edited by John T. Wixted. Copyright 2018 John Wiley & Sons, Inc]. Typically, Braille reading represents a low-tech vision substitution means allowing a subject to “read with the skin”. More technological advanced systems have since emerged. The Tactile Visual Substitution System (TVSS) has been introduced, to convert a tactile stimulation of the skin into visual information. It uses an array of 400 tiny tactile stimulators that transmit information (on the back of the subject) captured by a video camera. Another system using tactile to visual information conversion is the BrainPort device. It uses electro-tactile impulses to stimulate receptors on the surface of the tongue via a flexible electrode array receiving input from a head-mounted video camera. Auditory systems provide a higher spatial acuity and ability for parallel processing and have been developed “to see through the ears”. The system called “vOICe” converts visual images from a camera into sounds by transforming each pixel into a sound. Also, a device able to “hear with the skin” has been developed for deaf people. This device, called Versatile Extra-Sensory Transducer (VEST) from NeoSensory Inc., consists of an array of small vibration motors integrated into a vest. Attached to the vest is a microphone that captures sounds from the environment. These sounds are translated into tactile sensations perceived by the subject via the vibration motors. Finally, somatosensory information that can be usually sensed by the skin, such as temperature, pressure and force, can be captured by sensors and transformed into visual or hearing cues to a subject. However, when using sensory substitution devices, sensory overload is to be avoided. As well, interferences with natural environmental senses are to be avoided.

Low-threshold mechanoreceptors (LTMRs) are special mechanosensitive primary sensory neurons that react to innocuous mechanical stimulation (touch sensation). These cutaneous sensory neurons may be classified as either Ab, Ad or C based on their cell body sizes, axon diameter, degree of myelination and axonal conduction velocities. In addition, their firing pattern to sustain mechanical stimuli is variable, ranging from slow (SA) to intermediate (IA) and to rapidly adapting (RA).

LTMRs associated cutaneous end-organs encode touch stimuli and this encoding is then integrated and processed within the central nervous system. Both hairy and hairless (also named non- hairy or glabrous) skin areas contain discrete sets of LTMRs and associated end-organs (also named endings), and these different sets of LTMRs detect specific tactile modalities (cf. Table 1 and Figure 1).

In glabrous skin, four types of LTMRs with fast conduction velocity (Ab LTMRs) have been defined, each with a distinct terminal morphology (“endings”) and tuning property: (i) Ab SA1 -LTMRs (also herein identified as SAI-LTMRs) innervate Merkel cells in the basal epidermis, (ii) Ab SA2- LTMRs (also herein identified as SAII-LTMRs) are hypothesized to terminate in Ruffini corpuscles in the dermis, (iii) Ab RAl -LTMRs (also herein identified as RAI-LTMRs) innervate Meissner’s corpuscles in dermal papillae, and (iv) Ab RA2-LTMRs (also herein identified as RAII-LTMRs) terminate in Pacinian corpuscles deep in the dermis.

In hairy skin, apart from the SA1-LTMR / Merkel cell complex (touch dome), hair follicles are innervated by LTMR termination collars located just below the level of sebaceous gland. Despite their differences in sensitivity and encoding’ ability, the 3 types of lanceolate-ending LTMRs have identical terminal structures [A. Zimmerman etal. The gentle touch receptors of mammalian skin. Science, 2014; 346(6212), 940-954] The below table 1 identifies LTMRs of the skin and their corresponding end-organs (from A. Zimmerman et al. The gentle touch receptors of mammalian skin. Science, 2014; 346(6212), 940-954; V.E. Ahraira et al. The sensory neurons of touch. Neuron (2013); 79(4), 10.1016).

Table 1:

Signal transduction

The conventional view is that primary afferents (i.e., also named primary sensory neurons or LTMRs) are the first to participate to the signal transduction cascade. These neurons have a unique pseudo-unipolar morphology, with a single process that bifurcates into two branches: a distal branch, which can be up to a meter long, that innervates peripheral tissues and a shorter branch that terminates centrally. For the trigeminal system, which covers the head and face, the somata (or cell bodies) of peripheral sensory neurons reside within the trigeminal ganglia and terminate in the medulla. For the rest of the body, the somata reside within the dorsal root ganglia (DRG) and terminate in the spinal dorsal hom or dorsal column nuclei in the medulla. In order for primary sensory neurons to respond to mechanical stimuli and initiate action potentials, they need specific molecular transducers that can be directly activated by physical energy [F. Moehring etal. Uncovering the Cells and Circuits of Touch in Normal and Pathological Settings. Neuron 100, October 24, 2018]

Non-neuronal cells in the periphery were found to contribute intimately with primary sensory neurons to relaying touch signals centrally. Numerous specialized non-neuronal end-organs in the skin sense different features of mechanical stimuli: (1) Merkel cells respond to sustained touch and pressure and aid in two-point discrimination; (2) Ruffini’s end-organs sense stretching of skin around objects and over joints; (3) Pacinian corpuscles sense fast vibrations and deep pressure; (4) Meissner’s corpuscles sense slow vibrations and changes in texture; and (5) hair follicles detect hair movement in response to very light touch, clothing and air currents. How touch is tuned at each neurite complex is likely due to a variety of factors including the distinct expression patterns and density of mechanotransduction channels in sensory endings that innervate the end-organ and/or the nature of the non-neuronal cells that comprise the structural ending. The response also depends on other factors such as the depth of the end- organs in the skin, the extent of terminal branching, and the types of surrounding non-neuronal cells [F. Moehring et al. Uncovering the Cells and Circuits of Touch in Normal and Pathological Settings. Neuron 100, October 24, 2018]

Sensory restoration or sensory substitution devices use the Peripheral Nervous System (PNS), by either directly stimulating the appropriate nerve fibers in the context of sensory restoration, typically using electrodes (with or without wireless connection) or by stimulating the receptors/nerves of different senses in the context of sensory substitution (for example mechanoreceptors of the skin or ears, photoreceptors of the eyes, chemoreceptors of the nose or tongue). However, directly stimulating the nerves using electrodes requires the stability of the interactions between the electrode(s) and neurons with time and an appropriate selectivity of the electrode allowing a relevant electrode / neurons interaction [Hannes P. Saal el al. Biomimetic approaches to bionic touch through a peripheral nerve interface. Neuropsychologia 79 (2015) 344-353] As such, electrodes - by design (due to their size, hardness, composition, etc.) - may not be the optimized tools for stimulating neurons. Also, while the stimulation of receptors of our senses is interesting for sensory substitution, these receptors are usually engaged during sensory substitution and are not available for other tasks (see for instance Braille to “see/read with its skin/fmgertips”).

Therefore, there is a need for new tools allowing efficient, ideally improved, neural coding (i.e., processing of information), to the benefice of people requiring sensory restoration or substitution.

SUMMARY OF THE INVENTION

Inventor herein provides a minimally invasive and highly efficient tool/system (i.e., with appropriate spatial and temporal resolutions) spectacularly improving neural coding. The herein described system creates spatiotemporal electrical patterns at the level of the peripheral nervous system which can be efficiently read by the brain and are able to restore a perception, for example touch perception, in a subject who is deprived of it, to substitute a perception means to another in a subject suffering of an altered perception (for example of an altered vision or an altered hearing), to enhance perception in a subject, and/or to create new perception means in a subject. The system of the invention also enables for the first time the brain of a subject, for example of a human subject, to perceive beyond the reality the subject is used to perceiving thanks to his senses.

Herein described is a system (A) comprising genetically modified cells (B) and a removable device (C). The genetically modified cells are cells of the dermis, cells of the epidermis and/or Low Threshold Mechanoreceptors (LTMRs), and are activable by a signal emitted by the removable device (C). The removable device (C) typically collects an input signal which is, optionally processed and, used to activate the genetically modified cells (B), the removable device being wearable by a subject.

In a particular aspect, herein described is a system (A) comprising genetically modified cells (B) and a removable device/means (C), wherein the genetically modified cells (B) are (i) Merkel cells, lamellar cells in Meissner corpuscles and/or lamellar cells in Pacinian corpuscles, (ii) LTMRs (Low Threshold Mechanoreceptors) or (iii) LTMRs and one or several cells selected from Merkel cells, lamellar cells in Meissner corpuscles and lamellar cells in Pacinian corpuscles, and the genetically modified cells are activable by a signal emitted by the removable device/means (C). The removable device (C) collects an input signal which is, optionally processed and, used to activate the genetically modified cells (B), the removable device being wearable by a subject. This system is, according to a particular aspect of the invention, used for sensory enhancement in a subject, or for creating new sensory means in a subject, said new sensory means allowing the perception of a physical signal, chemical signal and/or biological signal which is not perceived by a sense of the subject, for example by a human sense.

A particular herein described system (A) is a sensory restoration system, a sensory substitution system, a sensory enhancement system, or a new sensory perception system.

Also herein described according to another particular aspect of the invention, are genetically modified cells (also herein identified as “genetically modified cells (B)”), in particular genetically modified cells for use for touch sensory restoration in an amputee or in a bum victim, or for sensory substitution in a subject at least partially or totally deprived of taste, smell, hearing, balance and/or vision, when said genetically modified cells are activated by an external light source of energy, the cells being cells of the dermis, cells of the epidermis and/or LTMRs of the amputee, bum victim or subject, and being genetically modified to express an optical protein which is not naturally present in and/or around said cells of the amputee, bum victim or subject.

In a particular aspect, herein described are genetically modified Merkel cells, lamellar cells in Meissner corpuscles and/or lamellar cells in Pacinian corpuscles, modified to express an optical protein which is not naturally present in and/or around said cells in a subject, for use for touch sensory restoration in an amputee or in a bum victim when said genetically modified cells are activated by an external light source of energy.

In another particular aspect, herein described are genetically modified Merkel cells, lamellar cells in Meissner corpuscles, lamellar cells in Pacinian corpuscles and/or LTMRs, modified to express an optical protein which is not naturally present in and/or around said cells in a subject for use for sensory substitution in a subject at least partially deprived of taste, smell, hearing, balance and/or vision when said genetically modified cells are activated by an external light source of energy.

Further herein described is a composition for use for touch sensory restoration in an amputee or in a bum victim, or for sensory substitution in a subject at least partially, or totally, deprived of taste, smell, hearing, balance and/or vision, wherein the composition comprises genetically modified cells, opsin(s) and/or nucleic acid sequence(s) coding for one or several distinct opsin(s), wherein the genetically modified cells express, or the nucleic acid sequence(s) code for, an optical protein which is not naturally present in and/or around cells or the dermis, cells of the epidermis and /or LTMRs of the amputee, bum victim or subject who will receive the composition, and wherein the genetically modified cells are activated by an external light source of energy. In a preferred aspect, the composition is a liquid, in particular a tattoo ink, or a gel. DETAILED DESCRIPTION OF THE INVENTION

Inventor herein advantageously describes a system (A) comprising genetically modified cells (B) and a removable device (C). The genetically modified cells are cells of the dermis, cells of the epidermis and/or LTMRs, and are preferably not located at a biological area of a subject corresponding to fingertips, mouth, lips and foot soles. The genetically modified cells are activable by a signal emitted by the removable device (C). The removable device (C) collects an input signal which is, optionally processed and, used to activate the genetically modified cells (B), the removable device being preferably wearable by a subject.

A preferred herein described system (A) comprises genetically modified cells (B) and a removable device/means (C), wherein genetically modified cells (B) are i) Merkel cells, lamellar cells in Meissner corpuscles and/or lamellar cells in Pacinian corpuscles, ii) LTMRs (Low Threshold Mechanoreceptors), or iii) LTMRs and one or several cells selected from Merkel cells, lamellar cells in Meissner corpuscles and lamellar cells in Pacinian corpuscles, and the genetically modified cells are activable by a signal emitted by the removable device/means (C), and the removable device (C) collects an input signal which is, optionally processed and, used to activate the genetically modified cells (B), the removable device being wearable by a subject.

In the context of the present invention, the subject is a subject having a brain, typically an animal, in particular a mammal, preferably a human, whatever its age or sex and health status.

A particular subject is a subject suffering of an altered perception, i.e., a subject suffering of an altered perception related to the lack of functioning or to the malfunctioning of one or more of his senses, typically a mammalian subject who doesn’t see, hear, smell, taste, touch and/or balance, or who doesn’t see, hear, smell, taste, touch and/or balance correctly, for example a diseased subject (a patient).

In such subjects, the present invention is typically used for “sensory restoration”, i.e., to restore a subject’s body particular functionality or full functionalities of the subject’s body, or for “sensory substitution”, thereby allowing the substitution of a particular functionality of the subject’s body or full functionalities of the subject’s body.

Another subject is a healthy subject who wants to experience sensory enhancement (“feel more/feel better”), i.e., who wants to better perceive outside stimuli (still within the natural possibilities offered by his/her senses), or a subject who wants to experience new perception, i.e., who wants to perceive a reality beyond the reality accessible through senses. GENETICALLY MODIFIED CELLS: TEMPORAL AND SPATIAL RESOLUTIONS

The genetically modified cells are cells of the dermis, cells of the epidermis and/or LTMRs. Genetically modified cells may be keratinocytes, melanocytes, Merkel cells, Langerhans cells, fibroblasts, mast cells, macrophages, lymphocytes and/or platelets.

In a particular aspect, genetically modified cells are lamellar cells in Pacinian corpuscles and/or lamellar cells in Meissner corpuscles.

Preferred genetically modified cells are Merkel cells, lamellar cells in Pacinian corpuscles, and/or lamellar cells in Meissner corpuscles.

LTMRs may be SAI-LTMRs, SAII-LTMRs, RAI-LTMRs, RAII-LTMRs, Ad-LTMRs and/or C- LTMRs.

In a particular aspect of the description, the genetically modified cells used in the context of the present invention are keratinocytes, melanocytes, Merkel cells, Langerhans cells, fibroblasts, mast cells, macrophages, lymphocytes and/or platelets, and/or LTMRs used in the context of the present invention are SAI-LTMRs, SAII-LTMRs, RAI-LTMRs, RAII-LTMRs, Ad-LTMRs and/or C -LTMRs.

In a particular aspect of the description, the genetically modified cells used in the context of the present invention are Merkel cells, lamellar cells in Pacinian corpuscles, lamellar cells in Meissner corpuscles, and/or LTMRs as herein described.

In the context of the present invention, a nucleic acid encoding an inhibitory (hyperpolarizing) or an excitatory (depolarizing) microbial opsin, or the inhibitory or excitatory protein itself, can advantageously be used to modify the cells.

In a particular aspect of the invention, a nucleic acid is used to “genetically modify” a cell for it to express the product of interest encoded by the nucleic acid. The nucleic acid is typically carried by a vector. When used to genetically modify a cell, the nucleic acid is for example carried by a viral vector, by a vector consisting of a nucleic acid cassette, or by a vector consisting of a plasmid.

In another typical aspect of the invention, a protein of interest as herein described, in particular an optical protein such as an opsin, is used to “modify” a cell (B). The protein of interest is exhibited by the “modified cell” or is expressed in or around said “modified cell”.

In a preferred aspect, the protein of interest is an optical protein which is not naturally present in or around cells of the dermis, cells of the epidermis and/or LTMRs of a subject, in particular an optical protein which is not naturally present in or around Merkel cells, lamellar cells in the Pacinian corpuscles, lamellar cells in the Meissner corpuscles and/or LTMRs of a subject.

In another preferred aspect, the protein of interest is expressed in and/or around modified cells which are not located at a biological area of a subject corresponding to fingertips, mouth, lips and foot soles. In a typical example of the invention, the inhibitory (hyperpolarizing) microbial opsin is a channel rhodopsin (ChR) or a variant thereof such as iChloC, SwiChRca, Phobos or Aurora; an halorhodopsin such as eNpHR, eNpHR2.0, eNpHR3.0, Halo or Jaws; or an archaerhodopsin such as eArch3.0, eArchT3.0 or ArchT.

In another typical example of the invention, the optical protein is an excitatory (depolarizing) microbial opsin, in particular a channel rhodopsin (ChR) or a variant thereof selected from TsChR, PsChR, ChR2, VChR2, ChRl, ChEF, VChRl, ReachR, MChR, C1V1, Chrimson, ChR2 H134R, CatCh (ChR2 L132C), ChR2 T159C, CatCh+ (ChR2 L132C-T159C), CheTA_A (ChR2 E123A), ChETA_TC (ChR2 E123T-T159C), ChETAx (ChR2 E123T), C1V1_T (ChRl/VChRl E126T), CV1_TT (ChRl/VChRl E122T-E162T), Chronos, Step-fimction opsins (ChR2 C128A/S/T; ChR2 D156A/C/N), ChlEF, Cs- Chrimson, vf-Chrimson and Slow ChloC (ChR2 E90R-D156N-T159C) [cf. J. Mattis et al. Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins. Nat Methods. 2014, 9(2): 159-172 ; T. Mager et al. High frequency neural spiking and auditory signaling by ultrafast red-shifted optogenetics. Nature Communications. 2018, 9:1750]

In a preferred aspect, an excitatory (depolarizing) microbial opsin as herein described, or a nucleic acid encoding such an opsin, is selected for use in the context of the invention.

In another preferred aspect, a fast channel rhodopsin (ChR) having a fast photocycle activity, typically presenting a fast deactivation time constant (fast closing kinetics) upon cessation of light, or a nucleic acid encoding such an opsin, is selected for use in the context of the invention. In this aspect, ChRs presenting a fast deactivation time constant below about 15 ms, below about 10 ms, below about 9 ms, below about 8 ms, below about 7 ms, below about 6 ms, or below about 5 ms, are particularly preferred. The following ChR variants are more particularly preferred: ChlEF, ChcTA_A. Chronos, and vf-Chrimson.

A single nucleic acid encoding an opsin, or several nucleic acids encoding distinct opsins, can be selected to genetically modify cells of a subject.

In a particular aspect, several nucleic acids encoding distinct opsins, typically at least two or more distinct opsins, selected for example among opsins presenting fast photocycle activity, opsins presenting low photocycle activities, opsins presenting specific peak activation wavelength (for example a blue peak activation wavelength, a green peak activation wavelength, a yellow peak activation wavelength or a red peak activation wavelength), and step-function opsins, can be used successively or simultaneously depending on the cell type which is to be modified, i.e. depending on the genetically modified cells which are to be stimulated/activated. Typically, at least two or more nucleic acids encoding different ChR variants can be used simultaneously during the activation of targeted cells and/or nerve fibers. The cells are considered to be “modified” when an optical stimulation triggers an electrical recording at the site of administration, typically at the site of injection, (“on state”), such an electrical recording being impossible in the absence of optical stimulation (“off state”).

Preferred genetically modified cells of the invention are for use for touch sensory restoration in an amputee or in a bum victim, or for sensory substitution in a subject at least partially deprived of taste, smell, hearing, balance and/or vision when said genetically modified cells are activated by an external light source of energy.

In a preferred aspect, the depth of the needle insertion when used to administer the genetically modified cells, opsin(s) and/or nucleic acid sequence(s) coding for one or several distinct opsin(s), is that observed in the context of tattoo procedure, i.e., the insertion is non-invasive and is not considered as a physical intervention on the human or animal body.

Inventors also herein describe a composition for use for touch sensory restoration in an amputee or in a bum victim, or for sensory substitution in a subject at least partially deprived of taste, smell, hearing, balance and/or vision, wherein the composition comprises genetically modified cells, opsin(s) or and/or nucleic acid sequence(s) coding for one or several distinct opsin(s), and wherein the genetically modified cells express, or the nucleic acid sequence(s) code for, an optical protein which is not naturally present in and/or around cells of the dermis, cells of the epidermis and/or LTMRs of an amputee, bum victim or subject, and wherein the genetically modified cells are activated by an external light source of energy.

As explained herein above, preferred genetically modified cells (B) express an excitatory (depolarizing) microbial opsin typically a channel rhodopsin (ChR) or a variant thereof such as TsChR, PsChR, ChR2, VChR2, ChRl, ChEF, VChRl, ReachR, MChR, C1V1, Chrimson, ChR2 H134R, CatCh (ChR2 L132C), ChR2 T159C, CatCh+ (ChR2 L132C-T159C), CheTA_A (ChR2 E123A), ChETA_TC (ChR2 E123T-T159C), ChETAx (ChR2 E123T), C1V1_T (ChRl/VChRl E126T), CV1_TT (ChRl/VChRl E122T-E162T), Chronos, Step-function opsins (ChR2 C128A/S/T; ChR2 D156A/C/N), ChlEF, Cs- Chrimson, vf-Chrimson or Slow ChloC (ChR2 E90R-D156N-T159C). These cells are preferably Merkel cells, lamellar cells in Pacinian corpuscles, lamellar cells in Meissner corpuscles and/or LTMRs, even more preferably Merkel cells, lamellar cells in Pacinian corpuscles, and/or lamellar cells in Meissner corpuscles.

In a preferred aspect of the description, the composition herein described is a composition comprising genetically modified cells, one or several distinct opsin(s) and/or nucleic acid sequence(s) coding for one or several opsin(s), and the composition is preferably a liquid, in particular a tattoo ink, or a gel. In a particular example, the composition is in the form of a liquid that turns into a gel once administered to a subject. In another aspect, the composition comprising genetically modified cells, one or several distinct opsin(s) and/or nucleic acid sequence(s) coding for one or several opsin(s), is part of a needle, a microneedle or a tip of a needle or microneedle.

FORMULATION OF GENETICALLY MODIFIED CELLS, OPSINS AND NUCLEIC ACIDS AND ADMINISTRATION

In a particular aspect, inventors herein describe the in vitro or ex vivo use of a genetic tool to (genetically) modify cells of a subject which are cells of the dermis, cells of the epidermis and/or LTMRs, preferably Merkel cells, lamellar cells in Pacinian corpuscles, lamellar cells in Meissner corpuscles and/or LTMRs, even more preferably Merkel cells, lamellar cells in Pacinian corpuscles and/or lamellar cells in Meissner corpuscles. Genetically modified cells typically express an optical protein which is not naturally present in and/or around said cells. In a preferred aspect, these cells are not located at a biological area of the subject corresponding to fingertips, mouth, lips and foot soles. Such a genetic tool is typically a nucleic acid construction, for example a nucleic acid cassette or a plasmid. The nucleic acid may also be included in a vector such as a plasmid or a viral vector. In the context of the present invention, genetic tools comprise opsin(s), nucleic acid sequence(s), vector(s), such as viral vector(s), and/or promoter(s).

Inventors also herein describe a method for modifying, for example for genetically modifying, in vivo, in vitro or ex vivo cells which are typically cells from the dermis, cells of the epidermis and/or LTMRs (such as the cells herein above identified), as well as the genetically modified cells obtained thanks to such a method.

The cells can be genetically modified using any method known to those skilled in the art which can be implemented in vitro, ex vivo or in vivo [cf. for example M. J. Benskey et al. Chapter 1. Basic Concepts in Viral Vector-Mediated Gene Therapy. Fredric P. Manfredsson and Matthew J. Benskey (eds.), Viral Vectors for Gene Therapy: Methods and Protocols, Methods in Molecular Biology, vol. 1937] Genetically modified cells obtained in vitro or ex vivo as well as herein described nucleic acid sequence(s), are typically formulated in a liquid or in a gel, in particular in a tattoo ink, or in a gel, before being administered to/implanted in, typically injected to, the subject. In a preferred aspect, the genetically modified cells obtained in vitro or ex vivo as well as herein described nucleic acid sequence(s), are part of a needle, a microneedle or of a tip of a needle or microneedle.

In a particular aspect, genetically modified cells and/or nucleic acid sequence(s) are formulated in a liquid that turns into a gel when administered in vivo, in the dermis up to the epidermis of a subject, typically at a biological area of the subject which is not a biological area corresponding to fingertips, mouth, lips and foot soles. Alternatively, cells can also be modified in vivo for example via a method comprising a step of administering a vector such as a plasmid or a viral vector [for example an adeno-associated virus (AAV) or a retrovirus such as a lentivirus] coding for one or several opsins, in the dermis up to the epidermis of a subject, typically at biological area of the subject which is not a biological area corresponding to fingertips, mouth, lips and foot soles. In a preferred aspect, cells are (i) Merkel cells, lamellar cells in Pacinian corpuscle and/or lamellar cells in Meissner corpuscles, (ii) LTMRs or (iii) LTMRs and one or several cells selected from Merkel cells, lamellar cells in Pacinian corpuscle and lamellar cells in Meissner corpuscles, and said cells are genetically modified in vivo.

The vector can be formulated in a liquid or in a gel, in particular in a liquid that turns into a gel when administered in vivo. In another and preferred aspect, the vector is part of a needle, of a microneedle or of a tip of a needle or microneedle.

When the transition from a liquid to a gel is triggered by a change of temperature, the liquid-to- gel transition typically occurs between 30°C and 40°C. Poly(D,L-lactic acid-co-glycolic acid)-6- poly(ethylene glycol)-ft-poly(D,L-lactic acid-co-glycolic acid) (PLGA-PEG-PLGA) triblock copolymers typically are materials which exhibit a sol-gel transition upon heating. The liquid to gel transition temperature is typically affected by the following parameters: the concentration of copolymer, the chain length of PEG, the chain length of PLGA, the molar ratio between PEG and PLGA, or the lactic acid/glycolic acid (LA:GA) ratio within the PLGA. All these parameters can be easily adjusted by the skilled person to trigger a liquid-to-gel transition at a temperature typically comprised between 30°C and 40°C, for example at the human body temperature.

When using a liquid that turns into a gel when administered to a subject, a controlled release of the genetically modified cells or of the nucleic acid sequence(s) (as such or in a vectorized form) at the site of administration can be obtained by an adaptation of the gel according to methods well-known by the skilled person in the art. Depending on the affinity between the gel and the genetically modified cells or nucleic acid sequence(s), a controlled release of the genetically modified cells or nucleic acid sequence(s) can be obtained, which lasts typically between few seconds (for example about two seconds) and 1 week. Alternatively, a controlled release of the genetically modified cells or nucleic acid sequence(s) lasting typically between 1 hour and 1 week, can be obtained depending on the kinetic of degradation of the selected gel.

The affinity between the gel and the genetically modified cells or nucleic acid sequence(s) typically depends on the nature of the bonds linking the genetically modified cells or nucleic acid sequence(s) and the material constituting the gel. The bond can be an hydrogen bond or be the result of electrostatic interactions.

The degradation of the gel typically consists in the swelling (i.e., expansion) of the gel or the breaking of bonds in the material(s) constituting the gel. The gel is ideally biodegradable. A biodegradable gel can typically comprises hydrolytic degradable polyesters blocks, such as poly(s- caprolactone) (PCL) blocks and poly(D,L-lactide-co-glycolide) (PLGA), blocks. Alternatively, the biodegradable gel can comprise polymer blocks with enzymatically degradable peptides, such as poly(L- alanine) (PA) blocks and chitosan blocks.

The genetically modified cells or nucleic acid sequence(s) (B) (as such or in a vectorized form) or composition comprising such genetically modified cells or nucleic acid sequences can be directly administered using typically a syringe and a needle when cells or nucleic acid sequence(s) are in suspension (i.e., when they are formulated as a liquid or as a gel, provided the viscosity of the gel remains compatible with such administration mode, for example as a liquid that turns into a gel when administered in a subject). The herein described genetically modified cells, opsins, nucleic acid sequence(s) or vectors can also be deposited on the surface of the skin and be spontaneously absorbed by the skin.

Alternatively, direct administration of opsin(s) or of nucleic acid sequence(s) encoding such opsin(s) (for example plasmid(s) encoding one or several distinct opsins) into targeted cells, typically cells of the dermis, cells of the epidermis and/or LTMRs can be performed in the context of the present invention.

As indicated herein above, the cells are preferably keratinocytes, melanocytes, Merkel cells, Langerhans cells, fibroblasts, mast cells, macrophages, lymphocytes, and/or platelets, and/or the LTMRs are preferably SAI-LTMRs, SAII-LTMRs, RAI-LTMRs, RAII-LTMRs, Ad-LTMRs and/or C -LTMRs. In a preferred aspect, the cells are lamellar cells within Pacinian corpuscles and lamellar cells within Meissner corpuscles.

In a preferred aspect, the targeted cells in which direct administration of opsin(s) or of nucleic acid sequence(s) encoding such opsin(s) (for example plasmid(s) encoding one or several distinct opsins) is(are) performed, are (i) Merkel cells, lamellar cells in Pacinian corpuscles and/or lamellar cells in Meissner corpuscles, (ii) LTMRs or (iii) LTMRs and one or several cells selected from Merkel cells, lamellar cells in Pacinian corpuscles and lamellar cells in Meissner corpuscles.

Several well-known technics involving biochemical or physical means, including lipofection, electroporation, optoporation or microinjection, may be used for direct administration.

The genetically modified cells or nucleic acid sequence(s) (as such or in a vectorized form) (B) or composition comprising such genetically modified cells or nucleic acid sequences can become the principal component of the needle(s), microneedle(s), or of the tip(s) of the needle(s) or microneedle(s). In such case, the needle(s), microneedle(s), or the tip(s) of the needle(s) or microneedle(s) is(are) inserted in vivo and remain(s) there. The erosion (such as degradation or dissolution) of the needle(s), microneedle(s) or of the tip(s) of the needle(s) or microneedle(s) triggers the release of the genetically modified cells or nucleic acid sequence(s), typically within seconds (for example about 2 seconds), hours, days or weeks following needle(s) or microneedle(s) insertion/implantation. The needle(s) or microneedle(s) are left in the skin for a selected period and can be removed at any time by extracting the part(s) of the needle(s) or microneedle(s) that has/have not been dissolved. Dissolvable needle(s) or microneedle(s) or dissolvable tip(s) of the needle(s) or microneedle(s) typically comprise(s) water soluble polymers, such as polyvinyl alcohol, polyvinylpyrrolidone or polyvinyl acetate, sugars, or any mixture thereof. The dissolvable needle(s) or microneedle(s) or tip(s) of needle(s) or microneedle(s) comprise(s) the herein described genetically modified cells or nucleic acid sequence(s).

In a preferred aspect, needle insertion is that observed in the context of tattoo procedure, i.e., insertion is non-invasive and is not considered as a physical intervention on the human or animal body.

Inventors also herein describe a composition for use for touch sensory restoration in an amputee or in a bum victim, or for sensory substitution in a subject at least partially deprived of taste, smell, hearing, balance and/or vision, wherein the composition comprises genetically modified cells, opsin(s) and/or nucleic acid sequence(s) coding for one or several distinct opsin(s), and wherein the genetically modified cells express an optical protein which is not naturally expressed by (present in and/or around) cells of the dermis, cells of the epidermis and/or LTMRs of the amputee, bum victim or subject, and wherein the genetically modified cells are activated by an external light source of energy.

In a preferred embodiment, the composition is a liquid, in particular a tattoo ink, or a gel. In a particular example, the composition is in the form of a liquid that turns into a gel once administered to a subject.

STABLE INTERACTION BETWEEN THE DEVICE (C) AND GENETICALLY MODIFIED CELLS (B)

In a preferred aspect of the invention, the removable device (C) advantageously stably interacts (in particular during the activation/stimulation step(s)) with genetically modified cells of a subject.

In another preferred aspect, both the removable device (C) and genetically modified cells (B) are not located at a biological area of the subject corresponding to fingertips, mouth, lips and foot soles. This is to limit, ideally avoid, interference with critical sensory biological area of human body. In other words, the removable device (C) and genetically modified cells (B) are localized/present on a biological area which is distinct of fingertips, mouth, lips and foot soles, and the removable device (C) and genetically modified cells (B) are preferably advantageously stably interacting together (in particular during the activation/stimulation step(s)).

The genetically modified cells (B) can be at multiple biological areas of a subject, typically 2, 3, 4, 5, 6, 7, 8, 9 or 10 different biological areas, which are preferably distinct of mouth, lips, fingertips and foot soles. The surface of a biological area represents typically about 0.5 cm², about 1 cm², about 2 cm², about 3 cm², about 4 cm², about 5 cm², about 6 cm², about 7 cm², about 8 cm², about 9 cm², about 10 cm², about 15 cm², or about 20 cm² of the skin of the subject.

Multiple sites/locations/spots of genetically modified cells (targeting multiple locations/sites/spots at one or multiple biological areas), typically more than one site/location/spot and up to typically 1000 locations/sites/spots of genetically modified cells at one or multiple biological areas (each area comprising one or multiple spots) of the subject are typically performed.

In other words, multiple sites/locations/spots of genetically modified cells (at one or several locations/sites/spots) may exist per biological area. For example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 400 or 500 locations/sites/spots of genetically modified cells per biological area exist.

When multiple sites/locations/spots of genetically modified cells exist per biological area, the distance between two adjacent spots is typically of less than about 100 pm, or of about 100 pm, about 200 pm, about 300 pm, about 400 pm, about 500 pm, about 600 pm, about 700 pm, about 800 pm, about 900 pm or about 1000 pm.

Typical biological areas where the genetically modified cells can be valuably present are for example areas where the subject is used to wear jewelry (such as a ring, a bracelet, a necklace) such as for example the subject’s arm, leg or ankle.

In a particular aspect of the invention, genetically modified cells, opsins, or nucleic acid sequence(s), typically vectors, in particular viral vectors, coding for one or several opsins, are to be administered once on a given site of implantation where they will ensure reproducible electrical signal/information transmission. In another particular aspect of the invention, genetically modified cells, opsins, or nucleic acid sequence(s), typically vectors, in particular viral vectors, coding for one or several opsins are to be administered several times on a given site of implantation. Repeated/successive administrations of genetically modified cells, opsins, or nucleic acid sequence(s), typically vectors, in particular viral vectors, coding for one or several opsins on a given site can typically be performed in order to increase the number of injection spots to the subject, the already genetically modified cells being still usable (i.e., used as “permanent” and “re-usable” neural interfaces).

Thus, the present invention advantageously allows spatiotemporal control of the stimulation of primary afferents (i.e., also identified as primary sensory neurons or LTMRs) through activation of the genetically modified cells.

In a particular aspect of the invention, neural data of a subject, such as data obtained from BOLD signal using functional Magnetic Resonance Imaging (fMRI) or electroencephalography (EEG) signals, may be recorded to assess the efficacy of neural coding in the subject. Neural data can also be recorded at the peripheral nerve system level (typically via a sensor, such as electrode (s)). These neural data may then be used as a feedback loop in order to “train” the system of the invention and/or to “train” the subject him/herself. Indeed, the information transmitted to the brain thanks to the system of the invention can be recorded (on a memory), processed by a processor, and then decoded (using the recorded/processed neural data and typically machine learning for neural decoding). The processed data can then advantageously be sent back to the subject in the form a signal perceivable by any one of the five natural senses of the subject in order to accelerate the learning process and facilitate any sensory restoration process, sensory substitution process, sensory enhancement process, or new sensory perception process. In a particular aspect, the decoded information obtained from a given subject can be used by said subject (i.e., transmitted to the subject and perceived by the subject) to facilitate learning and exploitation of information. Alternatively, the decoded information can be sent to the removable device (C) to update and improve output signal transmission.

These records can be used as herein above described thanks to the stable interaction existing (under activation) between the removable device (C) and the genetically modified cells (B).

In an aspect of the invention, such record(s) may be used to lock the system (A) of the invention by specifically associating the removable device (C) of the invention with the genetically modified cells (B) of the invention.

DEVICE

In a preferred aspect of the invention, the removable device (C) comprises a collector module (cl) the function of which is to collect an input signal. The input signal is typically selected from a physical signal, a chemical signal and a biological signal, and is used to activate the genetically modified cells (B). The collector module (cl) is capable of processing the signal when required.

A particular removable device (C) of the invention thus collects an input signal which is, optionally processed and, used to activate the genetically modified cells (B). The removable device is preferably wearable by a subject.

A typical device (C) of the invention comprises a collector module (cl) collecting an input signal which is typically a physical signal, a chemical signal or a biological signal, and capable of processing the signal when required, and a stimulator module (c2).

In a particular aspect, the collector module (cl) comprises a module (cl ) collecting an input signal and a processing module (cl”) encoding/converting the input signal into an output signal readable by the stimulator module (c2).

In another typical aspect, the stimulator module (c2) comprises a light source of energy, said source using the output signal to activate the genetically modified cells (B). As taught herein above, the collector module (cl) collects a signal which is typically a physical signal, a chemical signal or a biological signal or several signals, e.g., physical, chemical and/or biological signals.

A physical signal is for example an electromagnetic signal (see Figure 2) such as a radio wave signal, a microwave signal, a visible light signal, an infrared signal, an ultraviolet (UV) signal, an X-ray signal, a gamma-ray signal, etc.; a thermal radiation/heat signal; an electric signal; a magnetic signal; or a mechanic signal such as for example an ultrasound signal, a pressure signal or a strain signal.

The collector module can be a sensor module.

It is typically a “physical sensor module”, i.e., a sensor module collecting a physical phenomenon (i.e., a physical signal).

A physical sensor module can be an “image sensor module” detecting information in the form of light. An image sensor module typically consists of integrated circuits that sense the information and convert it into an equivalent current or voltage which can be later converted into digital data.

A physical sensor module can also be an “ultrasonic sensor module”. An ultrasonic sensor module is typically used to measure the distances between the sensor and an obstacle object. The ultrasonic sensor module generally works on the principle of the Doppler Effect and includes an ultrasonic transmitter and a receiver. The ultrasonic transmitter transmits the signal in one direction and this transmitted signal is then reflected back whenever there is an obstacle and is received by the receiver. The total time required for the signal to be transmitted and then received back is generally used to calculate the distance between the ultrasonic sensor and the obstacle.

A physical sensor module or physical sensor can also be for example an “infrared” sensor module; a “tactile” sensor module; a “pressure” sensor module; a “strain” sensor module; a “temperature” sensor module; a “magnetic-based” sensor (magnetometer) module; an “optical” sensor module; an “acoustic- based” sensor module; a “gravity” sensor (accelerometer) module; an “angular rate” sensor (gyroscope) module or a “deep pressure” sensor (barometer) module.

The sensor module (cl) can also be a “chemical sensor module” or “chemical sensor”, i.e., a module collecting a chemical phenomenon (i.e., a chemical signal). When the sensor module is a chemical sensor module, it is typically a liquid or gas sensor module detecting the composition or the concentration of a chemical agent in a medium such as for example an organic molecule or an ion.

The sensor module (cl) can also be a “biological sensor module” or “biosensor”, i.e., a module collecting a biological phenomenon (i.e., a biological signal). When the sensor module is a biosensor, it typically detects the composition or concentration of a biological agent such as for example a protein, a nucleic acid, a cell, a bacterium or a virus, in a medium;

Each sensor module is capable of processing the signal (if and when required) and typically combines sensing, computation, communications and power means into a very small volume typically below 100 mm³, below 10 mm³, or even below 1 mm³. A sensor module or several sensor modules, typically two or three sensor modules, or even a network of sensor modules can be combined in the device to increase its sensing ability. For instance, an optical sensor module can be coupled with an ultrasound sensor module to increase its sensing ability.

The collector module collecting the input signal can also be any other suitable means capable of collecting an input signal from one or several sensors (physical, chemical and/or biological sensors), or from one or several computing systems, for example any data generated by a computing system and transmitted in the form of a digital electrical signal, the sensor(s) or computing system(s) being external to the device. The input signal received by the collector module can be any input signal sent by remote sensor(s) and/or remote computing system(s), through wired (such as for example a HDMI or USB connector) or wireless connection, preferentially via a wireless connection such as for example Bluetooth and WIFI.

The herein described system is, according to a particular aspect of the description, used for sensory enhancement in a subject, or for creating new sensory means in a subject, said new sensory means allowing the perception of a physical signal, chemical signal and/or biological signal which is not perceived by a sense of the subject, for example by a human sense.

The sensory restoration, sensory substitution, sensory enhancement, or new sensory perception system according to the invention, preferably comprises a sufficient number “X” of dimensions or parameters and, in each dimension, a sufficient level “N” of features to build a robust information that will create or re-create sensory perception. Coding signal (corresponding to the signal emitted by the light source of energy from the stimulator module) can typically have dimensions expressed for example as intra-signals frequency, inter-signals frequency, signals amplitude, signals intensity, signals waveform, signals repetition, signals repetition frequency, signals total time, and any combination thereof. Also, distinct genetically modified cells can be used as a dimension (or parameter) for neural coding. As well, in each of these dimensions, “N” features can be implemented to (i) reconstitute a well- known perception such as a sound, a melody, colors, hue and luminance of a landscape, distance and direction, etc. and/or to (ii) create a new perception such as for example infrared vision or ultrasound vision. Machine learning can typically be used by the skilled person for such neural coding, or for implementing neuronal network methods to handle, typically transmit and/or record, information.

When the collector module (cl) comprises a module (cU) collecting an input signal and a processing module (cl”) encoding/converting the input signal into an output signal readable by a stimulator module (c2), the processing module (cl”) preferably contains a deep learning framework determining the parameters required to generate an output signal readable by the stimulator module. Machine learning can typically be used to encode information and implement a neuronal network method or system capable of determining the required parameters. In one aspect, the module (cl’) transmits signals to the processing module (cl”) which uses ADC (Analog -to-Digital Converter) or an equivalent converter, to perform the digitization of (digitalize) the acquired analog signals and generate an output signal which is then sent to a stimulator module (c2). For instance, the signal processing can be an image analysis, a text analysis (i.e., data analysis) or a speech analysis. The signal is captured by a module (cl’) of the collector module and sent to a processing module (cl”), for example for color segmentation, radiance segmentation, hue segmentation, sentence segmentation or word segmentation. Then, the processing module converts this input signal into an output signal and sends it to the stimulator module (c2).

In another context, a sensor module can sense a change of a measured parameter using a module (c l ) and transfer the information corresponding to this change to a processing module (cl”) (which can typically be a microcontroller) that calculates and converts the change into an output signal (containing all information from the input signal) readable by a stimulator module (c2).

The herein described stimulator module (c2) of the system (A) of the invention comprises a light source of energy, said source using the signal to activate the genetically modified cells (B).

Each collector module encodes an input signal into an output signal readable by a stimulator module, typically encodes/converts the input signal into an output signal readable by a light source of energy. Typically, the output signal can be an electrical signal to be sent to a stimulator module (c2) comprising (micro)LEDs acting as a light source of energy to activate the genetically modified cells (B).

Several stimulator modules, typically two or three stimulator modules, or a network of stimulator modules, can be combined in the device to increase its sensing ability. Several devices can also be used in parallel to increase sensing ability.

The spikes, generated in response to input signal(s) from the collector module(s), confirm the successful reading of the output signal by the stimulator module(s) present in the system (A) as well as the successful stimulation of the genetically modified cells by the light source of energy and consecutive induced stimulation of the peripheral nerves which will then convey/transmit a signal to the central nervous system which it can interpret (cf. Figure 3). These spikes can be recorded as electrophysiological signals and observed or decoded.

The removable device (C) is preferably powered by an external source or by a battery which is part of the device.

WEARABLE DEVICE DESIGN

The wearable device (C) is typically included in a jewelry, in a clothing or in a medical device. When included in a jewelry, it may be included for example in a ring, in a bracelet or in a necklace. When included in a clothing it may be included for example in a tee-shirt, in a sweatshirt, in a sock, in a mitt or in a glove, provided that it delivers reliable external stimulation to the genetically modified cells present under the subject’s skin. When included in a medical device, it may be included for example in an artificial skin (for example an ‘electronic skin’), in a patch or in a bandage.

In a particular aspect, the device (C) is a bracelet, a ring, a necklace, an artificial skin, a patch, a bandage, a mitt or a glove.

As herein indicated, the stimulator module (c2) preferably comprises a light source of energy, said source using the output signal to activate the genetically modified cells (B).

When several stimulator modules (c2) and/or different light sources of energy (for example blue, green, yellow and/or red-light sources of energy) are used, the opsin(s) or nucleic acid sequence(s) coding for one or several distinct opsins, used to modify cells are to be selected accordingly as taught in the present description.

STIMULATION PARAMETERS

When the source of energy source used to activate the genetically modified cells is a light source, the light stimulation (i.e., the signal) wavelength is typically selected based on the nature of the opsin(s) or nucleic acid sequence(s) coding for one or several opsin(s), used to modify the cells, in order to optimize the conversion of the signal emitted by the light source into an electrical signal.

In a preferred aspect of the description:

- the light stimulation (i.e., the signal) irradiance rate is between 0.1 mW/mm² and 1000 mW/mm²,

- the light stimulation (i.e., the signal) frequency is between 1 Hz and 500 Hz,

- the light stimulation (i.e., the signal) pulse width is between 5 ps and 500 ms, and/or

- the light stimulation (i.e., the signal) waveform is a monophasic square waveform, a rectangle waveform or a triangle waveform.

APPLICATIONS

Inventors herein describe a system (typically the herein described “system (A)”) and its use for sensory enhancement in a subject, or for creating new sensory means in a subject, for example in a human being, allowing the subject to perceive in particular a physical signal, a chemical signal and/or a biological signal which are, or on the contrary which are not, perceived by the subject’s senses, for example by a human sense.

They also herein describe genetically modified cells (typically the herein described “genetically modified cells (B)”) for use for touch sensory restoration in an amputee or in a bum victim, or for sensory substitution in a subject at least partially or totally deprived of taste, smell, hearing, balance and/or vision, when genetically modified cells are cells of the dermis, cells of the epidermis, and/or UTMRs and express an optical protein which is not naturally present in and/or around cells of the dermis, cells of the epidermis or UTMRs, and when genetically modified cells and/or modified nerve fibers are activated by an external light source of energy.

SENSORY TOUCH RESTORATION

Only in the United States, about two (2) million persons are living with the loss of a limb. There is a high need for these subjects to recover the sense of touch. A natural sensory feedback through their prostheses is typically sought for these persons. The system of the present invention offers to the subjects exhibiting proper LTMRs and/or end-organs functioning a limb axon-like stimulation. The system has the advantage of being biocompatible and of remaining at the site of implantation. The system of the invention may be advantageously used to stimulate afferent sensory fibers and provide efficient sensory feedback.

Genetically modified Merkel cells, lamellar cells in Meissner corpuscles and/or lamellar cells in Pacinian corpuscles, modified to express an optical protein which is not naturally present in and/or around said cells in a subject, are herein described for use for touch sensory restoration in an amputee or in a bum victim when said genetically modified cells are activated by an external light source of energy.

SENSORY SUBSTITUTION

According to the WHO (World Health Organization), the number of people of all ages visually impaired was estimated to be 285 million in 2010, of whom 39 million were blind. Also, around 466 million people worldwide have disabling hearing loss, and 34 million of these are children. It is estimated that by 2050 over 900 million people will have disabling hearing loss. In view of these major global health issues, sensory substitution, typically through people’s skin, could be a possibility for these people to “see” and/or “hear” with their skin.

Genetically modified Merkel cells, lamellar cells in Meissner corpuscles, lamellar cells in Pacinian corpuscles and/or UTMRs, modified to express an optical protein which is not naturally present in and/or around said cells in a subject, are herein described for use for sensory substitution in a subject at least partially deprived of taste, smell, hearing, balance and/or vision when said genetically modified cells are activated by an external light source of energy. SENSORY ENHANCEMENT

The present invention now makes it possible to very significantly enhance, and even widen, the capacities of sensory perception offered to a subject, in particular to a human being, by its natural senses.

In the vision field, the present invention now allows a subject for example to beneficiate of a 360° vision, to see throughout the whole earth in real time (i.e., acquire remote vision), to see underwater, to acquire space vision and see for example activities and phenomena occurring at an atomic scale up to a visible scale in and outside our solar system to increase perception and understanding of the universe.

In the touch field, the present invention now allows a subject for example to perceive touch from another subject with whom he/she is not in physical contact with (remote touch sensation).

In the smell and/or taste fields, the present invention now allows a subject for example to perceive a noxious (odorless and tasteless by common sense) chemical or biological compound; or to perceive a biological change and typically be able to early diagnose a cancer or any other life-threatening disease from a biological, for example blood, sample of a subject, for example with the sense of smell.

In the hearing field, the present invention now allows a subject for example to hear distant or remote (selected) sounds.

In the field of data (i.e., any information received as input signals which do not result from sensory experience or which cannot be perceived by a natural sense), the present invention now allows a subject for example to facilitate and/or increase data acquisition and/or processing (treatment) throughout daily activities, in particular in the context of learning.

NEW SENSORY

Beyond sensory enhancement, new sensory could be acquired by a subject which would allow the subject to enlarge his/her perception of reality compared to the reality as perceived through his/her natural senses.

Non-limiting examples of new sensory perceptions include the access to vision outside the visible domain, such as for example in the U.V. and/or infrared domain; or the access to sounds beyond current hearing ability such as for example the access to ultrasounds.

The use of a system for sensory enhancement in a subject, or for creating new sensory means in a subject allowing the perception of physical, chemical and/or biological signals which are not perceived by a sense of the subject is herein described. This system typically comprises genetically modified cells (B) and a removable device/means (C). The genetically modified cells (B) are preferably (i) Merkel cells, lamellar cells in Meissner corpuscles and/or lamellar cells in Pacinian corpuscles, (ii) LTMRs or (iii) LTMRs and one or several cells selected from Merkel cells, lamellar cells in Meissner corpuscles and lamellar cells in Pacinian corpuscles, and the genetically modified cells are activable by a signal emitted by the removable device/means (C). The removable device (C) collects an input signal which is, optionally processed and, used to activate the genetically modified cells (B), the removable device being wearable by a subject.

Typically, sensory restoration, sensory substitution, sensory enhancement, or the creation of new sensory means find application in a wide range of fields/industries/domains, such as in healthcare (typically by restoring senses and/or by substituting senses), in services (typically by enhancing life by providing assistance to persons), in communication, in defense/security (typically by making it possible to see, feel (touch), hear before it is accessible to normal human perception), in Aerospatiale (typically by augmenting knowledge), in agriculture, in automotive, in transports, in gaming, in sport, in entertainment (for example by augmenting entertainments experiences in music, in cinema), etc..

The embodiments of the invention described above are intended to be merely exemplary, and those skilled in the art will recognize, or will be able to ascertain using no more than routine experimentation, numerous equivalents of herein described specific materials and compositions as well as equivalents of herein described methods or procedures. All such equivalents are considered to be within the scope of the invention and are encompassed by the appended claims.

LEGENDS TO THE FIGURES

Figure 1: Biological components of dermis and epidermis.

Epidermis (zone I). The epidermis comprises the stratum comeum (nonviable epidermis) layer, the stratum lucidum (viable epidermis) layer, the stratum granulosum (viable epidermis) layer, the stratum spinosum (viable epidermis) layer, and the stratum basal (viable epidermis) layer. The epidermis comprises the following biological cells: the keratinocytes which represent 95% of cells and are present in each layer, and the melanocytes, the Merkel cells, and the Langerhans cells which represent 5% of the remaining cells and are present in viable epidermis. The epidermis also comprises the following appendages: hairs (hairy skin), sweat glands, sebaceous glands and lipids.

Dermis (zone ID. The dermis comprises the following biological cells: fibroblasts, mast cells, macrophages, lymphocytes and platelets. The dermis also comprises the following appendages: collagen fibrils, elastic connective tissue, mucopolysaccharides, highly vascularized network, lymph vessels, sensory nerves/nerve fibers, free nerve endings, end-organs such as Pacinian corpuscles, Meissner corpuscles, Ruffini corpuscles and/or longitudinal lanceolate endings, hair follicles, sebaceous gland and sweat glands. Lamellar cells are within Pacinian corpuscles and within Meissner corpuscles. Figure 2: Electromagnetic signals from the electromagnetic spectrum showing the range of wavelengths and frequencies spanned by electromagnetic radiations.

Figure 3: System (A) comprising genetically modified cells (B) and a removable device (C).

The genetically modified cells are activated by a light signal emitted by the removable device (C). The removable device (C) collects an input signal which is, optionally processed and, used to activate the genetically modified cells (B), the removable device being wearable by a subject. The device (C) typically comprises: a collector module (cl) collecting an input signal which is selected from a physical signal, a chemical signal and/or a biological signal. The input signal may typically be a physical signal, a chemical signal and/or a biological signal perceived by our natural senses, or be a physical, chemical and/or biological signal which cannot be perceived by one of the five natural senses (such as an infrared signal, an ultrasound signal, etc.). The collector module may comprise a collector module (c l ) collecting an input signal and a processing module (cl”) encoding the input signal into an output signal readable by the stimulator module (c2); a stimulator module (c2) comprising a light source of energy, said source using the output signal to activate the genetically modified cells (B).

The spikes, generated in response to input signal(s) from the collector module, confirm the successful reading of the output signal by the stimulator module present in the system (A) as well as the successful stimulation of the genetically modified cells by the light source of energy used to stimulate the peripheral nerves which will then convey/transmit a signal to the central nervous system which it can interpret.

EXPERIMENTAL PART

Protocol

Each test treatment method comprises a step of administering the genetically modified cells, opsins and/or nucleic acid sequences, typically vectors such as viral vectors, coding for one or several opsin(s), as herein above described by inventors, in the dermis up to the epidermis at a biological area of a subject which is not located at a biological area corresponding to fingertips, mouth, lips and foot soles and a subsequent step comprising activation of the genetically modified cells by an external light source of energy. The genetically modified cells are preferably Merkel cells, lamellar cells in Pacinian corpuscles, lamellar cells in Meissner corpuscles and/or LTMRs.

The recording of a signal at the peripheral nervous system level or at the central nervous system level confirms the activation of the genetically modified cells and their action on the nervous system. Concretely, an output signal read by the stimulator module (c2) comprising the light source of energy is converted into a signal that stimulates the peripheral nerves. Then, the peripheral nerves convey the information to the brain for neural coding and touch sensory restoration, sensory substitution, sensory enhancement or new sensory perception.

Claims

1. A system (A) comprising genetically modified cells (B) and a removable device/means (C), wherein:

- genetically modified cells (B) are i) Merkel cells, lamellar cells in Meissner corpuscles and/or lamellar cells in Pacinian corpuscles, ii) LTMRs (Low Threshold Mechanoreceptors), or iii) LTMRs and one or several cells selected from Merkel cells, lamellar cells in Meissner corpuscles and lamellar cells in Pacinian corpuscles, and the genetically modified cells are activable by a signal emitted by the removable device/means (C), and

- the removable device (C) collects an input signal which is, optionally processed and, used to activate the genetically modified cells (B), the removable device being wearable by a subject.

2. The system according to claim 1, wherein LTMRs are SAI -LTMRs, SAII-LTMRs, RAI- LTMRs, RAII-LTMRs, Ad-LTMRs and/or C -LTMRs.

3. The system according to claim 1 or 2, wherein the removable device (C) is stably interacting with genetically modified cells (B) of the subject and, both the removable device (C) and the genetically modified cells (B) are not located at a biological area of the subject corresponding to fingertips, mouth, lips and foot soles.

4. The system according to anyone of claims 1 to 3, wherein the genetically modified cells express an optical protein which is not naturally present in and/or around Merkel cells, lamellar cells in Meissner corpuscles, lamellar cells in Pacinian corpuscles and/or LTMRs of the subject.

5. The system according to claim 4, wherein the optical protein is a channel-rhodopsin or a variant thereof selected from TsChR, PsChR, ChR2, VChR2, ChRl, ChEF, VChRl, ReachR, MChR, C1V1, Chrimson, ChR2 H134R, CatCh (ChR2 L132C), ChR2 T159C, CatCh+ (ChR2 L132C- T159C), CheTA_A (ChR2 E123A), ChETA_TC (ChR2 E123T-T159C), ChETAx (ChR2 E123T), ClVlx (ChRl/VChRl E126T), CV1_TT (ChRl VChRl E122T-E162T), Chronos, Step-function opsins (ChR2 C128A/S/T; ChR2 D156A/C/N), ChlEF, Cs-Chrimson, vf-Chrimson and Slow ChloC (ChR2 E90R-D156N-T159C).

6. The system according to anyone of claims 1 to 5, wherein the device (C) comprises a collector module (cl) collecting an input signal which is selected from a physical signal, a chemical signal and a biological signal, the collector module (cl) being capable of processing the signal when required, and a stimulator module (c2).

7. The system according to claim 6, wherein the collector module (cl) comprises a module (cl’) collecting an input signal and a processing module (cl”) encoding the input signal into an output signal readable by the stimulator module (c2).

8. The system according to claim 6 or 7, wherein the stimulator module (c2) comprises a light source of energy said source using the output signal to activate the genetically modified cells (B).

9. The system according to anyone of claims 6 to 8, wherein the device (C) is included in a jewellery, in a clothing or in a medical device.

10. The system according to claim 9, wherein the device (C) is a bracelet, a ring, a necklace, an artificial skin, a patch, a bandage, a mitt or a glove.

11. Use of a system according to anyone of claims 1 to 10 for sensory enhancement in a subject, or for creating new sensory means in a subject allowing the perception of physical, chemical and/or biological signals which are not perceived by a sense of the subject.

14. Genetically modified Merkel cells, lamellar cells in Meissner corpuscles and/or lamellar cells in Pacinian corpuscles, modified to express an optical protein which is not naturally present in and/or around said cells in a subject, for use for touch sensory restoration in an amputee or in a bum victim when said genetically modified cells are activated by an external light source of energy.

15. Genetically modified Merkel cells, lamellar cells in Meissner corpuscles, lamellar cells in Pacinian corpuscles and/or LTMRs, modified to express an optical protein which is not naturally present in and/or around said cells in a subject for use for sensory substitution in a subject at least partially deprived of taste, smell, hearing, balance and/or vision when said genetically modified cells are activated by an external light source of energy.