CN106861050A - A kind of wireless neuromodulation device based on light science of heredity - Google Patents

Abstract

The present invention relates to medical device technology, in particular to a wireless neuromodulation device based on optogenetics, including a virus vector carrying a promoter, a light-sensitive protein and a fluorescent protein, and the promoter is used to specifically express light in a specific target tissue. Sensitive protein, to achieve specific and precise control, light-sensitive protein is the key to play a role; also includes LED light-emitting module and wireless control device; LED light-emitting module includes LED array and energy receiving system; wireless control device includes energy emission system and controller; The energy transmitting system transmits energy to the energy receiving system; both the LED array and the energy receiving system are encapsulated in a biocompatible insulating material, and are arranged close to the target tissue where the virus vector is located. The device realizes the wireless energy supply and remote control of the optogenetic stimulation system. It has the characteristics of miniaturization, wireless control, less heat generation, and less damage to target tissues, and the device can be used in fields where optogenetics can be applied.

Description

A wireless neuromodulation device based on optogenetics

technical field

The invention belongs to the technical field of medical devices, in particular to a wireless neuromodulation device based on optogenetics.

Background technique

Optogenetics is a rapidly developing bioengineering technology that integrates optics, software control, gene manipulation technology, electrophysiology and other multidisciplinary disciplines. The main principle is to firstly use genetic manipulation technology to transfer light-sensitive genes into specific cells to express light-sensitive proteins. Light-sensitive proteins will be selective for the passage of cations or anions under the light stimulation of corresponding wavelengths, resulting in The membrane potential inside and outside the cell membrane changes to achieve the purpose of selectively exciting or inhibiting cells, so as to achieve precise regulation of the physiological functions of cells, tissues, organs and organisms. Optogenetic technology has developed rapidly in neuroscience and other fields. In 2010, this technology was selected as the 2010 annual method of "Nature Methods" magazine, and is considered to be one of the most promising technologies to win the Nobel Prize in Biology.

Although the autonomic nervous system is not controlled by consciousness, it plays an important role in maintaining the life and health of the body. For example, changes in breathing, heart rate, and blood pressure in different states are controlled by the autonomic nervous system. The sympathetic and vagal components of the autonomic nervous system are antagonistic to each other, and the balance between the two is very important to maintain the health of the body. After a lot of basic and clinical research, it is found that the imbalance of the autonomic nervous system of the heart, especially the overexcitation of the sympathetic nerve, participates in and mediates the occurrence and development of various diseases such as arrhythmia, heart failure, myocardial infarction, and hypertension. Therefore, it is very important to regulate the autonomic nerves by various means to rebalance them. At present, the methods for rebalancing the autonomic nerves of the heart are mostly damage methods such as ablation and resection, which are prone to side effects. Using optogenetics to regulate the autonomic nerves not only does not damage the nerve , and can be precise and controllable. However, most of the current optogenetic stimulation systems are wired and require an optical fiber to be connected to the outside of the body, which is difficult to promote clinically, and even animal experiments have limitations in their activities.

Contents of the invention

The purpose of the present invention is to provide an optogenetics stimulation system with wireless power supply, which can be connected to wifi and interact through APP, so as to realize wireless regulation.

In order to achieve the above object, the technical solution adopted by the present invention is: a wireless neuromodulation device based on optogenetics, including a virus vector, which carries a promoter, a light-sensitive protein and a fluorescent protein and is implanted into the target tissue. The promoter is used to specifically express a light-sensitive protein in a specific target tissue to achieve specific and precise control, and the fluorescent protein is used to confirm sufficient expression; it is characterized in that it also includes an LED light-emitting module and a wireless control device; the LED light-emitting module It includes an LED array and an energy receiving system; the wireless control device includes an energy emitting system and a controller; the energy emitting system transmits energy to the energy receiving system; the LED array and the energy receiving system are packaged in a biocompatible In the insulating material, it is placed close to the target tissue where the viral vector is located.

In the above-mentioned wireless neuromodulation device based on optogenetics, the LED array includes several LEDs; the energy receiving system includes multiple antennas arranged around the several LEDs, and the multiple antennas are connected to the rectifier in turn and an amplifier; the amplifier connects several LEDs.

In the optogenetics-based wireless neuromodulation device described above, the energy transmission system includes a power supply system, an inverter, a control module, and an RF transmission unit connected in sequence; the controller includes a control panel connected to each other, a wifi receiving Module, integrated chip, and APP built in the program; the energy emission system is connected with the controller and packaged in the same shell.

In the optogenetics-based wireless neuromodulation device above, the control module is used to adjust stimulation parameters and emit matrix waves; the stimulation parameters include stimulation voltage, stimulation current, stimulation pulse width, intermittent time, and duration.

In the aforementioned wireless neuromodulation device based on optogenetics, the wireless neuromodulation device uses a built-in battery or an external power supply.

In the above optogenetics-based wireless neuromodulation device, the viral vector includes lentivirus and adeno-associated virus; the adeno-associated virus includes AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, Select the virus subtype according to the target tissue.

In the above optogenetics-based wireless neuromodulation device, the light-sensitive protein is selected from excitatory light-sensitive protein, inhibitory light-sensitive protein, and bidirectionally regulated light-sensitive protein.

In the aforementioned wireless neuromodulation device based on optogenetics, if ChR2 is selected as the excitatory light-sensitive protein, the LED array emits light with a wavelength of about 470nm; if VChR1 is selected, the LED array emits light with a wavelength of about 545nm If C1V1 is selected, the LED array emits light with a wavelength of about 540nm; if the inhibitory light-sensitive protein is ArchT, the LED array emits light with a wavelength of about 565nm; if eNpHR3.0 is selected, the The LED array emits light with a wavelength of about 590nm; if eBR is selected, the LED array emits light with a wavelength of about 540nm; if ChR2-step function opsins is selected for the bidirectionally regulated light-sensitive protein, the LED array emits light with a wavelength near 470nm The light is activated and suppressed by emitting light with a wavelength near 590nm; if VCHR-step function opsins are selected, the LED array emits light with a wavelength near 560nm for activation and emits light with a wavelength near 390nm for suppression.

In the above-mentioned optogenetics-based wireless neuromodulation device, the light-sensitive protein can be specifically expressed in the target tissue through a viral vector, or can be transformed into a target tissue through a transgenic animal model and the Cre-loxP system in animal experiments. Express light-sensitive protein.

Light-sensitive protein is the key to the function of wireless neuromodulation devices based on optogenetics. After being activated by light of a specific wavelength, the channel opens, causing ion flow inside and outside the cell membrane, changing the cell membrane potential, causing depolarization and hyperpolarization, thereby causing excitation and inhibition. effect.

The beneficial effect of the invention is that wireless energy supply and remote control of the optogenetic stimulation system are realized. It has the characteristics of miniaturization, wireless control, less heat generation, and less damage to target tissues. The device can be used in fields where optogenetics can be applied.

Description of drawings

Fig. 1 is the overall structure schematic diagram of an embodiment of the present invention;

Fig. 2 is a schematic diagram of an LED array and an energy receiving system according to an embodiment of the present invention;

Fig. 3 is a schematic diagram of a wireless control device and an energy transmission system according to an embodiment of the present invention;

Fig. 4 is a schematic diagram of connection between an energy transmitting system and an energy receiving system according to an embodiment of the present invention;

Fig. 5 is a schematic workflow diagram of an embodiment of the present invention;

Among them, 101-wireless control device, 102-radio frequency signal, 103-LED light-emitting module, 104-left stellate ganglion, 105-LED array, 106-LED emit light of specific wavelength, 201-energy emission system, 202- Energy receiving system and LED array, 401-viral vector injection schematic diagram, 402-viral vector, 403-light stimulation mechanism diagram.

detailed description

Embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings.

This embodiment adopts the following technical solution: a wireless neuromodulation device based on optogenetics, including a virus vector carrying a promoter, a light-sensitive protein and a fluorescent protein implanted in the target tissue, and the promoter is used to The light-sensitive protein is specifically expressed in a specific target tissue to achieve specific and precise control. The fluorescent protein is used to confirm sufficient expression; it is characterized in that it also includes an LED light-emitting module and a wireless control device; the LED light-emitting module includes LED arrays and energy The receiving system; the wireless control device includes an energy transmitting system and a controller; the energy transmitting system transmits energy to the energy receiving system; the LED array and the energy receiving system are all encapsulated in a biocompatible insulating material, set In close proximity to the target tissue where the viral vector is located.

Further, the LED array includes several LEDs; the energy receiving system includes multiple antennas arranged around the several LEDs, and the multiple antennas are sequentially connected to a rectifier and an amplifier; the amplifier is connected to several LEDs.

Further, the energy transmission system includes a power supply system, an inverter, a control module, and an RF transmission unit connected in sequence; the controller includes a control panel connected to each other, a wifi receiving module, an integrated chip, and a program built-in APP; the energy The launch system is connected to the controller and packaged in the same housing.

Further, the control module is used to adjust stimulation parameters and emit matrix waves; the stimulation parameters include stimulation voltage, stimulation current, stimulation pulse width, intermittent time, and duration.

Further, the wireless control device adopts a built-in battery or an external power supply.

Further, the viral vector includes lentivirus and adeno-associated virus; the adeno-associated virus includes AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, and the virus subtype is selected according to the target tissue.

Further, the light-sensitive protein is selected from excitatory light-sensitive protein, inhibitory light-sensitive protein, and bidirectionally regulated light-sensitive protein.

Further, if ChR2 is selected as the excitatory light-sensitive protein, the LED array emits light with a wavelength of about 470nm; if VChR1 is selected, the LED array emits light with a wavelength of about 545nm; if C1V1 is selected, the LED array Emit light with a wavelength of about 540nm; if the inhibitory photosensitive protein is selected as ArchT, the LED array will emit light with a wavelength of about 565nm; if eNpHR3.0 is selected, the LED array will emit light with a wavelength of about 590nm; if selected eBR, the LED array emits light with a wavelength of about 540nm; if ChR2-step function opsins are selected for the bidirectionally regulated light-sensitive protein, the LED array emits light with a wavelength near 470nm for activation, and emits light with a wavelength near 590nm for inhibition; VCHR-step function opsins, the LED array emits light with a wavelength near 560nm for activation, and emits light with a wavelength near 390nm for suppression.

Furthermore, the light-sensitive protein can be specifically expressed in the target tissue through a virus vector, or can be expressed in the target tissue through a transgenic animal model or Cre-loxP system in animal experiments.

During specific implementation, as shown in FIG. 1 , a wireless neuromodulation device based on optogenetics includes a virus vector, an LED light-emitting module and a wireless neuromodulation device. The virus vector enables the specific expression of light-sensitive proteins in the target area. The LED light-emitting module includes an LED array and an energy receiving system. The LED array is used to emit light suitable for the wavelength of the light-sensitive protein, and the energy receiving system is used to receive energy; System and controller; energy transmission system includes power supply system, inverter, control module and RF transmission unit; controller includes interconnected control panel, wifi receiving module, integrated chip, program built-in APP, energy transmission system and controller combined on one device and housed in the same housing. The record and storage of each stimulation is integrated into the chip, which can be connected to wifi, uploaded to APP, synchronized through mobile phones and other mobile terminals, and draw treatment curves.

As shown in Figure 2, the LED light-emitting module is used to emit light of a specific wavelength to stimulate the light-sensitive protein in the target tissue and change the potential of the inner and outer membranes of the target tissue cells, thereby exciting or inhibiting the target tissue. The LED light-emitting module contains several LED arrays and energy The receiving system is packaged in a biocompatible insulating material, and the LED array is close to the target tissue, which can fully irradiate each target tissue. The wavelength emitted by the LED depends on the light-sensitive protein. ArchT can be illuminated with a wavelength of about 565nm, and ChR2 can be illuminated with a wavelength of about 470nm. The energy receiving system includes an antenna around the LED, a rectifier and an amplifier; the antenna can receive the radio frequency signal sent by the wireless control device, which is converted into an electrical signal, and converted into direct current through the rectifier, and then amplified by the amplifier to generate enough energy for the LED to emit light . The energy receiving system is packaged with the LED array in a biocompatible insulating material.

As shown in Figure 3, the wireless control device includes an energy transmission system and a controller. The energy transmission system is composed of power supply system, inverter, control module and RF transmission unit; the controller is composed of control panel, wifi receiving module and APP. The wireless control device is powered by a built-in battery or an external power supply. The energy transmitting unit generates current and converts it into alternating current through an inverter. The alternating current passes through the RF transmitting unit to send out a radio frequency signal and transmit energy for the energy receiving system to receive. Its control module can adjust stimulation parameters, including stimulation voltage, stimulation current, stimulation pulse width, intermittent time, duration, and emit matrix waves. The wireless control device is also equipped with a wifi receiving module, an integrated chip, and a built-in APP in the program, which can store the records of each stimulus and release, connect to wifi and synchronize with mobile devices such as mobile phones by uploading the APP, and draw the treatment curve.

The wireless neuromodulation device can be used to regulate the autonomic nervous system of the heart, treat arrhythmia, improve myocardial ischemia, heart failure, prevent sudden death and other diseases by rebalancing the autonomic nervous system, and can also be used to implant the neck and other parts to achieve the treatment of depression Symptoms and other effects, not only can be used for clinical, but also for animal experiments.

Viral vectors are mainly lentiviruses and adeno-associated viruses. Adeno-associated viruses include AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, and AAV9. Specific virus types and virus subtypes are selected according to the target objects and target tissues. Animal experiments 1. Adeno-associated virus can be used for short-term use, which is characterized by high transfection efficiency. Lentivirus can be used for long-term experiments and clinical use, which is characterized in that it can be integrated into the host genome and expressed stably. The AAV9 subtype of adeno-associated virus mainly has high affinity to the heart, central nervous system, muscle and other tissues, and the AAV8 subtype mainly has high affinity to the liver, eye, and nervous system, causing specific virus subtypes to be selected according to different target organs. Available AAV9 subtypes. Excitatory light-sensitive proteins, inhibitory light-sensitive proteins, and bidirectionally-regulated light-sensitive proteins are used as needed to inhibit the activity of stellate ganglia and treat arrhythmias. ArchT and eNpHR can be used as inhibitory light-sensitive proteins, and the virus vector also carries specificity. The promoter expresses the target gene only in the target organ to achieve specific and precise control. In animal experiments, transgenic animal models and the Cre-loxP system can also be used to express light-sensitive proteins, and only the LED lighting system and wireless energy control and control system of the present invention are used.

The LED array is used to emit light of a specific wavelength, stimulate light-sensitive proteins in the target tissue, change the potential of the inner and outer membranes of the target tissue cells, and thereby excite or inhibit the target tissue. The LED array contains several LEDs packaged in a biocompatible insulating material, close to The target tissue can be fully irradiated to the entire target tissue. The wavelength emitted by the LED depends on the light-sensitive protein. ChR2 uses light with a wavelength of about 470nm; VChR1 uses light with a wavelength of about 545nm; C1V1 uses light with a wavelength of about 540nm; ArchT uses light with a wavelength of about 565nm; eNpHR3.0 uses light with a wavelength of about 590nm; Light with a wavelength of about 540nm is used.

The wireless neuromodulation device of this embodiment can not only be used to regulate the autonomic nerves of the heart, but also can be used to control the central nervous system and the cervical vagus nerve to treat depression. Due to its miniaturization and wireless regulation, it produces less heat and damages the target tissue. Small features, the system can be used in the fields where optogenetics can be applied.

In the following, the wireless neuromodulation device of this embodiment is applied to reduce ventricular arrhythmia and improve the prognosis of myocardial ischemia by inhibiting the left stellate ganglion. As shown in Figure 1, the AAV9 virus carrying ArchT is first surgically injected into the left stellate ganglion 104, and an LED light-emitting module 103 is implanted nearby. After about 4 weeks, ArchT is stably expressed, and at this time, the wireless regulation is turned on. The device 101 controls the switch, and adjusts the stimulation parameters in the control panel. The wireless control device 101 sends out a radio frequency signal 102, which is received by the implanted LED light emitting module 103. The energy receiving system inside the LED light emitting module 103 receives the signal, and a current appears to supply The LED array 105 emits 565nm wavelength light 106 to irradiate the left stellate ganglion 104, and the ArchT in the left stellate ganglion 104 is activated, resulting in changes in the distribution of ions inside and outside the nerve cells, hyperpolarized changes in the membrane potential, and inhibition of nerve electrical activity. Thereby reducing the excitability of the sympathetic nerve and reducing the incidence of ventricular arrhythmia after myocardial infarction.

A schematic diagram of the LED array 104 irradiating the left stellate ganglion as shown in FIG. 2 . The antenna in the energy receiving system 202 inside the LED light emitting module 103 receives the radio frequency signal, converts it into electrical energy, and converts it into direct current through a rectifier, and amplifies it through an amplifier to supply LED light.

As shown in Figure 3, the energy transmission system 201 in the wireless control device 101 includes a power supply system, an inverter, a control module and an RF transmission unit, and has a built-in integrated chip and a wifi module, and a built-in APP. The energy is supplied by a built-in battery or an external power supply, and is converted into alternating current through an inverter, and the current flows through the RF transmitting unit, which emits a wireless radio frequency signal to power the LED.

As shown in FIG. 4 , the wireless energy emitted by the energy transmitting system 201 is received by the energy receiving system 202 for supplying the LED to emit light.

As shown in FIG. 5 , a schematic diagram 401 of viral vector injection, and a schematic diagram 403 of light stimulation mechanism. The viral vector 402 carrying a specific sequence, the promoter function is to specifically express in a specific target tissue, and the opsin, that is, the light-sensitive protein, plays the role of an ion channel, which is the core of the system. Inhibitory or sensitive types can be selected according to needs. Fluorescent proteins can be used to detect whether the target protein is expressed. In this embodiment, AAV9 can carry ArchT as a light-sensitive protein and connect to a fluorescent protein. After the virus is injected into the left stellate ganglion of 104, the virus is stably expressed after about 4 weeks. At this time, 565 nm light is given to the left shape ganglion ( LSG), which changes the inner and outer membrane potential of nerve cells, hyperpolarizes nerve cells, and inhibits the nerve activity in LSG to reduce the occurrence of arrhythmia after myocardial infarction.

It should be understood that the parts not described in detail in this specification belong to the prior art.

Although the specific embodiments of the present invention have been described above in conjunction with the accompanying drawings, those of ordinary skill in the art should understand that these are only examples, and various variations or modifications can be made to these embodiments without departing from the principles and principles of the present invention. substance. The scope of the invention is limited only by the appended claims.

Claims (9)

1. a kind of wireless neuromodulation device based on light science of heredity, including viral vectors, the viral vectors carrying promoter, Photaesthesia albumen and fluorescin implantation destination organization, the promoter are used in the specific expressed photaesthesia of specific objective tissue Albumen, realizes that specificity is precisely controlled, and the fluorescin gives full expression to for confirmation；Characterized in that, also luminous including LED Module and wireless regulation and control device；The LED light module includes LED array and energy acceptance system；The wireless regulation and control device Including energy emission system and controller；The energy emission system transmits energy to the energy acceptance system；It is described LED gusts Row and energy acceptance system are packaged in biocompatibility insulating materials, the target where being arranged on the viral vectors At tissue.

2. the wireless neuromodulation device of light science of heredity is based on as claimed in claim 1, it is characterised in that the LED array Including several LED；The energy acceptance system includes the multiple antennas being arranged on around several LED, the multiple antenna It is sequentially connected rectifier and amplifier；The amplifier connects several LED.

3. the wireless neuromodulation device of light science of heredity is based on as claimed in claim 1, it is characterised in that the energy transmitting System includes the electric power system, inverter, control module and the RF transmitter units that are sequentially connected；The controller includes being connected with each other Control panel, wifi receiver modules, integrated chip, the built-in APP of program；The energy emission system is connected and seals with controller In same shell.

4. the wireless neuromodulation device of light science of heredity is based on as claimed in claim 3, it is characterised in that the control module For adjusting stimulation parameter, matrix ripple is sent；When the stimulation parameter includes stimulation voltage, stimulating current, stimulates pulsewidth, interval Between, the duration.

5. the wireless neuromodulation device of light science of heredity is based on as claimed in claim 1, it is characterised in that the wireless regulation and control Device uses internal battery or external power supply.

6. the wireless neuromodulation device of light science of heredity is based on as claimed in claim 1, it is characterised in that the viral vectors Including slow virus and adeno-associated virus；The adeno-associated virus include AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, virus subtype is selected according to destination organization.

7. the wireless neuromodulation device of light science of heredity is based on as claimed in claim 1, it is characterised in that the photaesthesia egg It is white to select excitability photaesthesia albumen, inhibition photaesthesia albumen, bidirectional modulation photaesthesia albumen.

8. the wireless neuromodulation device of light science of heredity is based on as claimed in claim 7, it is characterised in that the excitability light If sensitive Protein launches the light of 470nm or so wavelength from ChR2, the LED array；If from VChR1, the LED The light of array emitter 545nm or so wavelength；If from C1V1, the light of the LED array transmitting 540nm or so wavelength； If the inhibition photaesthesia albumen launches the light of 565nm or so wavelength from ArchT, the LED array；If from ENpHR3.0, the then light of LED array transmitting 590nm or so wavelength；If from eBR, the LED array transmitting The light of 540nm or so wavelength；If the bidirectional modulation photaesthesia albumen is from ChR2-step function opsins, The LED array transmitting 470nm wavelength light activation nearby, nearby wavelength light suppresses transmitting 590nm；If from VCHR- Step function opsins, then the LED array transmitting 560nm wavelength light activation nearby, wavelength near transmitting 390nm Light suppresses.

9. the wireless neuromodulation device of light science of heredity is based on as claimed in claim 1, it is characterised in that the photaesthesia egg Bai Jike by viral vectors it is specific expressed in targeted tissue, can also pass through in zoopery transgenic animal model, Cre-loxP systems make destination organization express photaesthesia albumen.