METHODS TO REGULATE POLARIZATION AND ENHANCE FUNCTION OF CELLS

• US 20150283265A1
• Filed: 06/17/2015
• Published: 10/08/2015
• Est. Priority Date: 08/05/2005
• Status: Active Grant

• Current Assignee: Gholam A. Peyman
• Original Assignee: Gholam A. Peyman
• Inventors: Peyman, Gholam A.
• Granted Patent: US 10,022,457 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61B 2017/00345 Micromachines, nanomachines, microsystems
• CPC Class Codes: A61B 5/1114 Tracking parts of the body
• CPC Class Codes: A61B 5/24 Detecting, measuring or recording bioelectric or biomagnetic signals of the body or parts thereof
• CPC Class Codes: A61B 5/6867 specially adapted to be attached or implanted in a specific body part A61B5/6847 takes precedence
• CPC Class Codes: A61F 9/0079 using non-laser electromagnetic radiation, e.g. non-coherent light or microwaves
• CPC Class Codes: A61F 9/008 using laser
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 38/465 acting on ester bonds (3.1), e.g. lipases, ribonucleases
• CPC Class Codes: A61K 41/0042 Photocleavage of drugs in vivo, e.g. cleavage of photolabile linkers in vivo by UV radiation for releasing the pharmacologically-active agent from the administered agent; photothrombosis or photoocclusion
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/6923 the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
• CPC Class Codes: A61K 47/6929 the form being a nanoparticle, e.g. an immuno-nanoparticle
• CPC Class Codes: A61K 48/0041 the non-active part being polymeric
• CPC Class Codes: A61K 49/0067 quantum dots, fluorescent nanocrystals
• CPC Class Codes: A61K 9/0009 involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis microelectromechanical systems A61K9/0097
• CPC Class Codes: A61N 2005/063 comprising light transmitting means, e.g. optical fibres
• CPC Class Codes: A61N 2005/0651 Diodes
• CPC Class Codes: A61N 2005/0659 infrared
• CPC Class Codes: A61N 2005/0661 ultraviolet
• CPC Class Codes: A61N 2005/0662 : Visible light
• CPC Class Codes: A61N 5/0601 : Apparatus for use inside the body
• CPC Class Codes: A61N 5/0613 : Apparatus adapted for a specific treatment
• CPC Class Codes: A61N 5/062 : Photodynamic therapy, i.e. excitation of an agent
• CPC Class Codes: A61N 5/0622 : Optical stimulation for exciting neural tissue
• CPC Class Codes: A61N 5/067 : using laser light
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 5/00 : Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Abstract

Methods and compositions to controllably regulate cells at a target site. A quantum dot-targeting agent complex is administered to a patient in need of therapy, and the complex is stimulated using an implanted fiber optic system. In embodiments, the system includes an electrical sensor that detects and monitors electrical activity of the stimulated controllably regulated cells, and relays this information to a controller that can regulate further stimulation.

Claims

1. A method for at least one of editing or regulating a gene in a target excitable cell, the method comprising

administering to a patient in need thereof a plurality of nanoparticles coated with a biocompatible molecule for cell uptake, the nanoparticles conjugated with at least one opsin-family gene, an antibody that targets the nanoparticles to an excitable cell, and a CRISPR/cas9 complex forming a complex of coated nanoparticle-gene-CRISPR/cas9,

stimulating the complex with an energy source under conditions sufficient to introduce the gene into the target cell and, using the CRISPR/cas9 complex, to modify a pathology-inducing DNA in the cell to treat the patient.

2. The method of claim 1 where the DNA is selected from the group consisting of nuclear DNA and mitochondrial DNA.

3. The method of claim 1 where the CRISPR/cas9 complex excises a DNA segment bearing a pathology-inducing mutation in the opsin-family gene.

4. The method of claim 3 further comprising replacing the excised DNA segment bearing a pathology-inducing mutation in the opsin-family gene with an unmutated DNA segment in the opsin-family gene using the CRISPER/cas9 complex.

5. The method of claim 1 where the excitable cell is selected from the group consisting of a retinal cell, a central nervous system neuronal cell, a central nervous system glial cell, a spinal cord cell, a peripheral nerve cell, a cardiac cell, and combinations thereof.

6. The method of claim 1 where the complex is administered systemically or locally.

7. The method of claim 1 where the complex is administered to a site selected from the group consisting of intraocular, intravitreal, intrathecal, brain ventricle, over a brain, proximate a central nervous system neuron, proximate a peripheral nerve, or intracardiac.

8. The method of claim 1 where the patient has a neurodegenerative disease or a cardiac rhythm disease.

9. The method of claim 1 further comprising, before administering the complex of coated nanoparticle-gene-CRISPR/cas9 to the patient, administering the complex of coated nanoparticle conjugated-gene-CRISPR/cas9 to in vitro cultured patient stem cells.

10. The method of claim 9 where the patient stem cells are selected from the group consisting of conjunctival limbal stem cells, heart stem cells, brain stem cells, spinal cord nerve stem cells, mesenchymal stem cells, ectodermal stem cells, and combinations thereof.

11. The method of claim 1 where the nanoparticles are piezoelectric nanoparticles and the stimulating step is controlled remotely to wirelessly stimulate the piezoelectric nanoparticles and produce an electrical current created by a wireless electrical stimulation of cells resulting in altering a membrane potential or action potential of cells in contact with the piezoelectric nanoparticles.

12. The method of claim 11 resulting in a cellular membrane response selected from the group consisting of hyperpolarization, hypopolarization, and depolarization.

13. The method of claim 11 where stimulation uses an ultrasound probe.

14. The method of claim 11 where the piezoelectric nanoparticles are administered a route selected from the group consisting of injecting submucosally, spraying a nasal mucosa, and combinations thereof, where the piezoelectric nanoparticles travel through olfactory nerves to the brain that is stimulated with an externally applied ultrasound over the nose by a patient as needed.

15. The method of claim 11 where stimulation of the piezoelectric nanoparticles is controlled by a diode laser surgically implanted in the patient, and the piezoelectric nanoparticles act as an “

on”

/“

off”

switch of a battery supplying electricity to the diode laser.

16. The method of claim 15 where stimulation is controlled by programmable processor of an extra-corporal ultrasonic probe wirelessly controlled by the diode laser.

17. The method of claim 15 where the diode laser is surgically implanted at a site selected from the group consisting of external to the skull, internal to the skull, adjacent to an organ, adjacent to sympathetic ganglion cells, adjacent to parasympathetic ganglions cells, adjacent to the brain, adjacent to the heart, adjacent to an eye, adjacent to the spinal cord, adjacent to peripheral nerves, and combinations thereof.

18. The method of claim 14 where the patient has a pathology selected from the group consisting of epilepsy, migraine, depression, paresis, paralysis, spastic muscles numbness, nerve injuries, dementia, spinal cord injury, peripheral nerve injury, Parkinson'"'"'s disease, Alzheimer'"'"'s disease, restless leg syndrome, and combinations thereof.

19. The method of claim 11 where the piezoelectric nanoparticles are rendered biocompatible by coating with a compound selected from the group consisting of zirconate titanate, perovskite-based, oxides, barium titanate, polyvinylidene fluoride (PVDF), and combinations thereof.

20. The method of claim 16 where a ribbon of piezoelectric nanoparticles are attached to the battery of the diode that, upon ultrasound stimulation, creates a pulsating electrical current in the piezoelectric nanoparticles leading to the battery of the diode laser, thus controlling activation of the diode laser light production and initiating an “

on”

/“

off”

action potential in the membrane of excitable cells depending on the frequency and duration of the electrical pulse.

21. The method of claim 1 where the complex is injected into an excitable organ having a brain blood barrier, resulting in complex entry into the organ in the absence of degradation by enzymes in the blood.