Articles and methods for treating mitochondrial dysfunction

Abstract

Provided herein are methods of treating mitochondrial dysfunction. The methods include applying stimulation to target cells in a subject in need thereof, wherein the stimulation is configured to drive push respiration in the target cells.

Description

[0001] ARTICLES AND METHODS FOR TREATING MITOCHONDRIAL DYSFUNCTION

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] The present application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 63 / 669,848, filed July 11, 2024, which application is incorporated herein by reference in its entirety.

[0004] BACKGROUND OF THE INVENTION

[0005] Mitochondria play a critical role in cellular respiration, and are responsible for converting nutrients into adenosine triphosphate (ATP). A decline in mitochondrial function, such as through aging or other acquired mitochondrial dysfunction, can lead to a general decline in cellular function. Such decline can affect various tissues and organs, and is linked to various age- related diseases. For example, it is currently believed that aging is related to deficiencies in the mitochondrial electron transport chain (ETC) where high energy electrons are able to escape from the chain and combine with an oxygen molecule forming superoxide, which is a free radical (otherwise known as a reactive oxygen species). Free radicals donate the high energy electron that was intended for ATP generation within the ETC to other body tissues such as mitochondrial membrane lipids or proteins, causing damage to these structures. When enough damage occurs, a vicious downward spiral begins where the ETC becomes decoupled from ATP production because of electron leak, which itself is caused by free radical damage to the very membrane proteins and lipids which compose the inner mitochondrial membrane. This results in a tissue that has deficient energy (ATP) production due to damage to the very anatomical structures which are intended to produce the energy, leading to various diseases.

[0006] Accordingly, there remains a need in the art for articles and methods that improve on existing articles and methods for treating mitochondrial dysfunction. The present disclosure meets this need.

[0007] SUMMARY OF THE INVENTION

[0008] In one aspect, a method of treating mitochondrial dysfunction, the method including applying stimulation to target cells in a subject in need thereof; wherein the stimulation is configured to drive push respiration in the target cells. In some embodiments, the push respiration comprises forced production of adenosine triphosphate (ATP) in the target cells. In some embodiments, the stimulation includes magnetic stimulation. In some embodiments, the magnetic stimulation comprises transcranial magnetic stimulation (TMS). In some embodiments, the applying stimulation step comprises varying a stimulation frequency over time. In some embodiments, the applying stimulation step comprises varying the stimulation over time with relation to a cardiac cycle of the subject.

[0009] In some embodiments, the TMS is administered with at least one of lower power and increased frequency. In some embodiments, the lower power includes less than 260 Joules per pulse. In some embodiments, the treating includes cortical flux therapy treatment and the lower power includes between 25 and 100 Joules per pulse. In some embodiments, the treating includes deep white matter tract treatment and the lower power includes between 75 and 150 Joules per pulse. In some embodiments, the increased frequency includes greater than 20 Hz. In some embodiments, the increased frequency includes at least 30 Hz. In some embodiments, the increased frequency includes at least 50 Hz. In some embodiments, the TMS is administered for between 1 and 60 minutes. In some embodiments, the TMS is administered for between 5 and 30 minutes. In some embodiments, the TMS is administered for at least 5 minutes per hour.

[0010] In some embodiments, the target cells comprise cells experiencing mitochondrial dysfunction. In some embodiments, the target cells include glial cells.

[0011] In some embodiments, the subject has a condition involving mitochondrial dysfunction. In some embodiments, the condition includes at least one of acute acquired mitochondrial dysfunction, chronic acquired mitochondrial dysfunction, and congenital mitochondrial dysfunction. In some embodiments, the mitochondrial dysfunction is from at least one of reduced oxygenation and age. In some embodiments, the condition comprises ischemic stroke. In some embodiments, the stimulation is administered repeatedly for 3 days following the ischemic stroke.

[0012] In some embodiments, the method further includes obtaining diffusion weighted magnetic resonance imaging (MRI) data from the subject and integrating the MRI data into the administering of the stimulation. In some embodiments, the integrating of the MRI data includes interpolating three-dimensional coordinates from the MRI data into a treatment helmet. In some embodiments, the integrating provides coordination between the MRI data and the stimulation.

[0013] In some embodiments, a system for treating brain tissue includes a capacitor configured to store energy from an energy source; a pulse shaper configured to release the energy from the capacitor at specific intensities and intervals; and a treatment coil configured to receive the energy from the pulse shaper and deliver the energy to the brain tissue of a subject. In some embodiments, the capacitor comprises up to 160 microFarad at 1700 volts. In some embodiments, the capacitor comprises up to 120 microFarad at 1700 volts. In some embodiments, the system is configured to administer the method according to claim 1.

[0014] BRIEF DESCRIPTION OF THE DRAWINGS

[0015] For a fuller understanding of the nature and desired objects of the present invention, reference is made to the following detailed description taken in conjunction with the accompanying drawing figures wherein like reference characters denote corresponding parts throughout the several views.

[0016] FIG. 1 depicts a flowchart illustrating a method of administering transcranial magnetic stimulation (TMS) according to an embodiment of the disclosure.

[0017] FIG. 2 depicts a schematic illustrating a system for administering transcranial magnetic stimulation (TMS) according to an embodiment of the disclosure.

[0018] DETAILED DESCRIPTION OF THE INVENTION

[0019] Definitions

[0020] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described.

[0021] The articles “a,” “an,” and “the” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article, unless the context clearly dictates otherwise. By way of example, “an element” means one element or more than one element.

[0022] Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. “About” can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term about. As used herein, the terms “subject” and “patient” are used interchangeably. As used herein, a subject is preferably a mammal such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) and a primate (e.g., monkey and human), most preferably a human.

[0023] As used in the specification and claims, the terms “comprises,” “comprising,” “containing,” “having,” and the like can have the meaning ascribed to them in U.S. patent law and can mean “includes,” “including,” and the like.

[0024] Unless specifically stated or obvious from context, the term “or,” as used herein, is understood to be inclusive.

[0025] Ranges provided herein are understood to be merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range (as well as fractions thereof unless the context clearly dictates otherwise). For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges (e.g., from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc.), individual numbers within that range (1, 2, 3, 4, 5, 6), and fractions thereof (1.4, 2.7, 3.5, 4.2, 5.9, 6.1, etc.). This applies regardless of the breadth of the range.

[0026] Detailed Description

[0027] Methods

[0028] Provided herein are methods of treating mitochondrial dysfunction. In some embodiments, the method includes applying stimulation to target cells / tissue to deposit energy (i.e., adenosine triphosphate (ATP)) into the cells. This deposition of energy into the cells through applied stimulation is referred to herein as “push respiration.” Any suitable stimulation for depositing energy into the cells can be applied to the target cells / tissue. In some embodiments, the stimulation includes pulsed stimulation. In some embodiments, the stimulation includes magnetic stimulation, such as, but not limited to, transcranial magnetic stimulation (TMS). In some embodiments, the stimulation includes varying the direction vector of the magnetic field over time. In some such embodiments, varying the direction vector over time captures more mitochondrial ATPase proton channels. In some embodiments, the stimulation frequency is varied over time in order to optimally load mitochondria within the subject tissue and maximize the benefits of push respiration. In some embodiments, the stimulation is varied over time with relation to the cardiac cycle to force push respiration to occur during times of peak bloodflow or bloodflow nadir. In some embodiments, the stimulation is performed with a solenoidal coil surrounding a body part such as a joint and rapidly alternating the current within the solenoid in order to maximize differences in electromotive force within the tissue and its mitochondria over time. In some embodiments, the stimulation is applied such that the incident magnetic and electrical fields can be focused within deep tissue compartments and shaped in order to generate magnetic and electrical flux in all possible directions to maximize mitochondrial ATPase and ETC recruitment

[0029] In contrast to existing TMS therapy, which is focused on the effects of TMS on entire cells, the methods described herein directly impact intracellular organelles, such as mitochondria. For example, in the context of neural tissue, rather than targeting direct stimulation of neurons to action potential as with existing TMS applications, the methods disclosed herein utilize push respiration to deposit energy into any cells experiencing mitochondrial dysfunction. In some embodiments, push respiration powers the mitochondria to make ATP, even in unfavorable conditions. In some embodiments, push respiration increases metabolic activity, which occurs for a sustained period of time after one or more stimulation sessions. As will be understood by those skilled in the art, the duration of the sustained period of time will vary according to the amount of ATP produced / deposited in the cells, which can be controlled according to the parameters of the stimulation applied. For example, in some embodiments, the method is applied to deposit / produce a relatively smaller amount of ATP. In such embodiments, the sustained period of time will last up to 10 minutes following treatment, up to 5 minutes following treatment, up to 4 minutes following treatment, up to 3 minutes following treatment, up to 2 minutes following treatment, up to 1 minute following treatment, between 30 seconds and 10 minutes following treatment, between 1 minute and 10 minutes following treatment, or any suitable combination, sub-combination, range, or sub-range thereof. In some embodiments, the method is applied to deposit / produce a relatively larger amount of ATP. In such embodiments, the sustained period of time will last at least 10 minutes, at least 30 minutes, at least an hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 12 hours, at least 18 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, or any suitable combination, sub-combination, range, or sub-range thereof. Additionally or alternatively, in some embodiments, push respiration facilitates the synthesis and expression of complex proteins such as Brain Derived Neurotrophic Factor (BDNF), along with response to BDNF in a trophic manner.

[0030] In some embodiments, rather than treating neurons, as with existing applications of TMS, the methods disclosed herein primarily treat glial cells. Glial cells are the most common cells in the brain and support neurons in their roles. Glial cells insulate neurons with myelin, which allows neural axons to communicate complex information more effectively and efficiently. They are also much more hardy than neurons because they don’t consume as much oxygen, and supply energy to neurons, which have a very high metabolic activity. Finally, glial cells have “foot processes” which fill in all of the gaps in the inner layer of small brain arteries. The inner layer of any artery in the body is known as the endothelium. Outside of the brain, the endothelium is finely porous, allowing certain small molecules to pass freely from the blood vessels into the tissue. The brain, however, is too sensitive an organ to allow that to happen, and so those foot processes of the glial cells actually fill in all of those little gaps, forming what is known as the “Blood-Brain Barrier.” Accordingly, the methods disclosed herein are not limited to treatment of neural tissue and, in some embodiments, include treating glial cells and / or any other suitable brain tissue.

[0031] The stimulation according to the methods described herein can be applied with any suitable parameter for depositing energy in the target cells / tissue ( / .e., powering the mitochondria to make ATP). In some embodiments, the target cells / tissue can be treated through push respiration using stimulation with unexpectedly lower power and / or increased frequency as compared to existing stimulation paradigms. For example, in some embodiments, the method includes administering TMS to a subject in need thereof with a lower power and / or increased frequency as compared to existing TMS applications.

[0032] As used herein, the term “lower power” refers to a comparatively lower power than that used in existing stimulation (e.g., magnetic stimulation therapy, TMS therapy). Suitable lower power includes, but is not limited to, up to 260 Joules per pulse, up to 200 Joules, up to 150 Joules, up to 100 Joules, between 25 and 260 Joules, between 25 and 200 Joules, between 25 and 150 Joules, between 25 and 100 Joules, between 50 and 260 Joules, between 50 and 200 Joules, between 50 and 150 Joules, between 50 and 100 Joules, 75 and 260 Joules, between 75 and 200 Joules, between 75 and 150 Joules, between 75 and 100 Joules, or any suitable combination, sub- combination, range, or sub-range thereof. In some embodiments, the lower power range varies depending upon the specific tissue being treated. For example, in some embodiments, the lower power range for cortical flux therapy treatment includes between 25 and 100 Joules. In another example, the lower power range for deep white matter tract treatment includes between 75 to 150 Joules.

[0033] Additionally, as used herein, the term “increased frequency” refers to a comparatively higher frequency than that used in existing stimulation (e.g., magnetic stimulation therapy, TMS therapy). Suitable increased frequencies include, but are not limited to, greater than 20 Hz, at least 30 Hz, at least 50 Hz, up to 1,000 Hz, between 21 Hz and 1,000 Hz, between 25 Hz and 1,000 Hz, between 30 Hz and 1,000 Hz, between 50 Hz and 1,000 Hz, or any combination, subcombination, range, or sub-range thereof.

[0034] The lower power and / or increased frequency TMS may be administered for any amount of time suitable for treating a disease, injury, or condition. For example, suitable duration of TMS administration may include, but is not limited to, at least 1 minute, between 1 minute and 60 minutes, between 1 minute and 45 minutes, between 1 minute and 30 minutes, between 5 minutes and 60 minutes, between 5 minutes and 45 minutes, between 5 minutes and 30 minutes, or any suitable combination, sub-combination, range, or sub-range thereof. In some embodiments, the stimulation is administered for at least 5 minutes per hour. Alternatively, in some embodiments, the stimulation is administered for less than 5 minutes per hour, such as, but not limited to, 3-20 minutes per day. As will be appreciated by those skilled in the art, the duration of treatment may vary depending upon the disease, injury, or condition. In some embodiments, the TMS therapy described herein may be administered prophylactically to reduce, slow, or prevent disease and / or condition onset. Additionally or alternatively, in some embodiments, the TMS therapy described herein may be administered therapeutically.

[0035] As will be appreciated by those skilled in the art, the methods disclosed herein may be used to efficaciously treat a wide array of conditions involving mitochondrial dysfunction. In some embodiments, the conditions include acute acquired mitochondrial dysfunction, chronic acquired mitochondrial dysfunction, and / or congenital mitochondrial dysfunction.

[0036] In some embodiments, the acquired mitochondrial dysfunction is a result of reduced oxygenation. For example, the methods disclosed herein can be used to treat conditions resulting from injuries that reduce oxygenation to cells / tissue and / or occur in cells / tissue with naturally lower oxygenation. Injuries that reduce oxygenation include, but are not limited to, ischemic stroke; traumatic brain injury; acute arterial occlusion, stenosis, or vasospasm; chronic arterial occlusion or stenosis; acutely diminished tissue perfusion (shock) whether due to hemorrhage, sepsis, diminished cardiac output, or other factors; acute or chronic venous outflow obstruction; acute hypoxia due to pulmonary ventilation perfusion mismatch (pneumonia, pulmonary embolus); acute myocardial infarction; congestive heart failure; acute heart failure; hypoxic ischemic injury; acute limb ischemia; acute mesenteric ischemia; acute testicular or ovarian torsion; chronic limb ischemia; or any other injury that reduces oxygenation to cells / tissue. Other injuries that can be treated by the methods described herein include, but are not limited to, frostbite, white matter damage from high blood pressure / diabetes, and / or other trauma that results in mitochondrial dysfunction due to resources being diverted towards healing (e.g., post- traumatic healing). Cells / tissues with naturally lower oxygenation include, but are not limited to, cartilage, ligaments, or any other cells / tissue with comparatively lower blood supply.

[0037] In some embodiments, the methods disclosed herein can be used to treat conditions resulting from diseases involving decreased oxygenation / metabolism. Such diseases include, but are not limited to, conditions involving glial cell dysfunction (e.g., demyelinating diseases, multiple sclerosis, chronic ischemic injury (e.g., related to hypertension or diabetes), acute / chronic toxic / metabolic injury (e.g., alcoholism)), neurodegenerative diseases (e.g., Alzheimer’s disease, frontotemporal dementia, multiple system atrophy, Parkinson’s disease, Lewy Body dementia), and / or cancers (e.g., glioblastoma multiforme (GBM)). In some embodiments, the neurodegenerative disease includes non-infectious neurodegenerative diseases. In some embodiments, the neurodegenerative disease includes any non-infectious neurodegenerative disease with a mitochondrial dysfunction etiology. In some embodiments, the neurodegenerative disease includes early dementia and / or mild cognitive impairment, where treatment as described herein facilitates repair of cellular damage.

[0038] In some embodiments, the methods disclosed herein include repairing cellular damage within living cells. In some embodiments, the method includes therapeutically treating one or more of the conditions disclosed herein. For example, in some embodiments, the method includes therapeutically treating acute acquired mitochondrial dysfunction (e.g., ischemic stroke) by effecting push respiration according to one or more of the embodiments disclosed herein in a subject in need thereof. In some embodiments, the method includes prophylactically treating one or more conditions through push respiration. For example, in some embodiments, the method includes prophylactically treating aging through push respiration. Additionally or alternatively, in some embodiments, the method includes prophylactically treating Alzheimer’s disease. In some such embodiments, the prophylactic treatment of Alzheimer’s disease includes administering stimulation, according to one or more of the embodiments disclosed herein, to effect push respiration in a subject prior to onset of any symptoms (e.g., in early middle age). As will be appreciated by those skilled in the art, in some embodiments, the treatment described herein stops and / or delays the progression / worsening of a condition without reversing the condition. Alternatively, in some embodiments, the treatment partially or completely reverses the condition.

[0039] In some embodiments, the method further includes integrating volumetric TMS treatment (e.g., for stroke) with imaging obtained at diffusion weighted magnetic resonance imaging (MRI), which shows the location of the injured tissue and the distance from the skin surface. For example, in some embodiments, as illustrated in FIG. 1, the three dimensional coordinates from the diffusion weighted MRI are interpolated into a treatment device, providing coordination between the diagnostic imaging and the TMS treatment. In some embodiments, this process is automated at every level (z.c., from diagnosis to stimulation location to stimulation intensity and frequency). Device

[0040] Also provided herein are systems for treating brain tissue using transcranial magnetic stimulation (TMS). In some embodiments, the system 200 includes a TMS system. In some embodiments, the TMS system includes a capacitor (which stores charge), a pulse shaper (which determines the pulse duration, frequency, and power), and a treatment coil. Referring to FIG. 2, in some embodiments, during the administering of the TMS treatment energy from an energy source 201 (e.g., a wall outlet) is transmitted (e.g., along a wire) into a capacitor 203, where it is stored for future use. The energy is then released from the capacitor 203 via a pulse shaper at specified intensities and intervals. This energy released from the capacitor by the pulse shaper is transmitted (e.g., via wire) to a coil 205 positioned on or adjacent to the subject’s head, and is released in the form of electrical current. This current induces a magnetic field and that discharged magnetic energy results in a brief but powerful electrical field within the subject’s tissue which places electromotive force on all charged particles within the treatment region of the tissue. This electromotive force induces movement of said charged particles and is itself electrical energy, some of which can be absorbed and utilized by cellular and subcellular components of the subject’s tissue.

[0041] In some embodiments, the capacitor of the TMS system is smaller, less expensive, and / or lighter as compared to existing TMS systems currently utilized for depression. For example, in contrast to existing TMS systems, which have a 180 microFarad capacitor rated at 1700 Volts DC (providing a total amount of stored charge of about 0.3 Coulomb), the capacitor of the presently disclosed system, at 1700 V, may be up to 160 microFarad, up to 150 microFarad, up to 140 microFarad, up to 130 microFarad, up to 120 microFarad, between 100 and 160 microFarad, between 110 and 160 microFarad, between 120 and 160 microFarad, between 100 and 150 microFarad, between 110 and 150 microFarad, between 120 and 150 microFarad, or any suitable combination, sub-combination, range, or sub-range thereof.

[0042] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents are considered to be within the scope of this invention and covered by the claims appended hereto.

[0043] It is to be understood that wherever values and ranges are provided herein, all values and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present invention. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application.

[0044] The following examples further illustrate aspects of the present invention. However, they are in no way a limitation of the teachings or disclosure of the present invention as set forth herein.

[0045] EXPERIMENTAL EXAMPLES

[0046] Example 1 : Ischemic Stroke

[0047] In an acute ischemic stroke, a blood clot travels from the heart or body arteries into a brain blood vessel and gets lodged there. The blood flow does not typically drop to zero because other nearby blood vessels can help contribute to nutrient delivery and waste removal. But the oxygen level in the deprived tissue drops to very low levels, and there is not enough oxygen to sustain neurons. Neurons need a constant and high supply of oxygen to survive, and they begin to die 3 minutes after the blood clot gets lodged. By the time the patient reaches the hospital, most of the neurons in the affected region are dead.

[0048] The glial cells are not, however. They are much more hardy because they don’t consume as much oxygen as neurons. However typically within the 3 days that the artery stays clogged before the body’s normal clot breakdown mechanisms reopen the artery, many of the glial cells die too. Since the glial cells comprise the blood brain barrier, loss of the glial endothelial foot processes results in disruption of the blood brain barrier. Once the blood brain barrier is disrupted, the glial cells begin to die faster, and by the time the causative blood clot breaks down, the endothelium is so porous that entire blood cells begin to seep into the tissues. Those blood cells degrade, forming compounds which are toxic to the sensitive tissues, damaging them permanently.

[0049] The endothelium and glia form the skeleton of the brain, and once destroyed there is no getting it back. Therefore, by saving the glial cells and endothelium such that they remain intact for the first three days following stroke (“acute phase”), new progenitor neurons can implant into the viable tissue and rebuild the brain.

[0050] Even during the acute phase of the stroke, blood is getting into the tissue due to collateral blood supply. The carotid artery divides in the neck into two main branches. One which goes to the face, skin, scalp, orbits, and skull, the other which serves the forebrain, including the motor and sensory transduction strips and the entire prefrontal cortex. When a stroke to one of these regions occurs, blood supply can, in most individuals, shunt from the face and skull into the brain. It is much less than before, but it is not zero.

[0051] This reduced blood supply impacts cellular respiration, which is the process by which cells manufacture Adenosine Triphosphate (ATP). Cellular respiration occurs deep within cells within engines called mitochondria. The mitochondria is shaped like a pill capsule and it is composed of two membranes, one surrounding the other. The inner membrane is folded in many different dimensions in order to maximize surface area. Embedded within that inner membrane is a complex protein molecule which sticks out of either side called the FIFO ATP synthase (ATPase). That protein catalyzes a chemical reaction that attaches another phosphate ion to the end of Adenosine Diphosphate, which is the inactive form known as ADP. Once the new phosphate ion is added to ADP, ATP has been reformed. When any process is taking place inside cells, that process requires electrical energy. ATP sheds one of its phosphate ions, a chemical reaction which releases electrical energy into specific intracellular machinery which uses that energy to fuel and maintain the machinery of life. However it takes energy for the ATPase to make ATP from ADP. That energy comes in the form of electricity, specifically, the movement of positively charged protons across a proton gradient, established using another protein complex embedded in the inner mitochondrial membrane.

[0052] Also embedded within the inner mitochondrial membrane is this other group of proteins known as Complexes 1-4, collectively known as the electron transport chain. The electron transport chain works using the principle of redox reactions, or oxidation-reduction reactions. The electron is deposited into the top of complex 1 by NADH, which is produced from blood glucose in the Krebs Cycle within the center of the mitochondria, inside of both mitochondrial membranes, deep to the inner mitochondrial membrane. This space is known as the “Matrix.”

[0053] The electron is deposited into complex 1 where it is sticking out into the matrix. The electron comes from NADH, an earlier intermediary product of the first two stages of cellular respiration. Those are known as glycolysis and the citric acid cycle (also referred to as the Krebs Cycle). That electron is very high in energy. Once it gets deposited into complex 1, it gets sucked out of the matrix towards the intermembrane space by the next protein in the complex 1 electron transport chain membrane protein complex, because that next protein in the chain has slightly more affinity for the electron. That higher affinity forms a temporary bond between the electron and that protein, and a small amount of energy is released in the process. That same energy is then simultaneously used to pump protons from the matrix into the intermembrane space, the space between the inner and outer mitochondrial membranes. That causes a higher concentration of protons to form in the intermembrane space than in the matrix.

[0054] That concentration difference forms a gradient of charged particles which, by definition, is an electrical field. This electrical field contains potential energy, much like a battery or capacitor. That electrical field is then dissipated by protons fluxing through the ATPase, restoring an equal balance of protons across the inner mitochondrial membrane, producing ATP, the energy currency of life, as a byproduct.

[0055] At the end of the electron transport chain, oxygen is waiting as the final oxidizing agent. It combines with the electron permanently, forming water. All along this chain of membrane proteins, oxygen is gently pulling energy out of the food we eat, in the form of high energy electrons in motion. That is, movement of electrons = movement of charged particles = electricity. This whole process is referred to herein as “pull respiration,” which is fully dependent on oxygen delivery to tissues.

[0056] So in the setting of a stroke, a very delicate highly energy intensive organ suddenly becomes starved of oxygen. There isn’t as much oxygen around so there is less “pulling” taking place through the electron transport chain. Since there is less pulling, there are less electrons moving and they are moving slower, therefore the electrical output is diminished. That means fewer protons getting pulled from the matrix into the intermembrane space by the work of oxygen via the electron transport chain. That means a weaker gradient of protons fluxing from the intermembrane space back into the matrix via the ATPase. That means markedly lower energy production. Famine is established and cells are at risk of destruction because life is highly energy dependent.

[0057] Push respiration, on the other hand, is an entirely new concept that does not require oxygen delivery. Instead, an electrical gradient is supplied across a tissue using a TMS pulse. When a portion of the TMS electrical gradient vector aligns with the axis of an ATPase embedded in an inner mitochondrial membrane, that electrical gradient facilitates proton flux through the ATPase. The ATPase does not run in reverse, so the component of the electrical vector in line with the ATPase’ s proton vector will facilitate proton flux through the ATPase. This proton flux bypasses the electron transport chain, producing more ATP and adding to the cell’s energy reserves in the absence of oxygen. But the reverse is not true, so net energy deposition, in the form of ATP, into cells is possible via TMS, where the magnetic field generates an electrical gradient.

[0058] Accordingly, the glial cells and endothelium may be saved through push respiration by TMS according to the embodiments disclosed herein. Because push respiration is taking place at the subcellular mitochondrial level, and because the tissue of the brain most sensitive to oxygen deprivation, the brain cortex, is closest to the skin surface, less powerful pulses (e.g., 10-30% or 20-50% machine output) with high frequencies can be used. This establishes a near constant electrical flux through the brain’s outer layer, the cortex, which is the most sensitive to low oxygen because it is very energy dependent and sensitive to drops in energy production. This would also increase safety by decreasing vibration, and would be much more comfortable, allowing the subject to wear the device on or just above their head for as much time as possible during those first three days.

[0059] Additionally or alternatively, in order to get into the deeper white matter tracts affected by the stroke, higher power continuous theta burst protocols at 70-90% of motor threshold may be used. That is, nearly continuous low power high frequency flux therapy may be used for superficial cortical tissues, while interspersed higher power continuous theta burst sequences may be used for deeper tissue. The greater the volume of adequately treated brain, the greater volume of brain which will fully recover post stroke.

[0060] Example 2: Alzheimer’s Disease

[0061] While the pathophysiology of Alzheimer’s Disease is far more complex than that of acute ischemic stroke, it too is a disease of energy (ATP) deficiency. The current prevailing theory of Alzheimer’s Disease is that it is caused by similar mechanisms as aging itself, only in an accelerated form. The accelerated disease course is due to the high metabolic / energy demand of neuronal tissue.

[0062] The prevailing theory of aging is that it is related to deficiencies in the mitochondrial electron transport chain where high energy electrons are able to escape from the chain and combine with an oxygen molecule forming superoxide, which is a free radical (otherwise known as a reactive oxygen species). While this is a normal phenomenon, it occurs more frequently in highly metabolic tissue such as neuronal tissue. Free radicals donate the high energy electron that was intended for ATP generation within the ETC to other body tissues such as mitochondrial membrane lipids or proteins, causing damage to these structures. When enough damage occurs, a vicious downward spiral begins where the electron transport chain becomes decoupled from ATP production because of electron leak which itself is caused by free radical damage to the very membrane proteins and lipids which compose the inner mitochondrial membrane.

[0063] Thus we are left with a tissue that has deficient energy (ATP) production due to damage to the very anatomical structures which is intended to produce the energy, said damage caused by electron leakage and superoxide formation. Over decades, this progressive damage results in the characteristic pathology of Alzheimer’s Disease:

[0064] 1) extracellular insoluble amyloid Beta plaques; and

[0065] 2) intracellular tau protein aggregates known as neurofibrillary tangles which destabilize neuronal microtubules essential for transmission of metabolic and signaling molecules from the cell body into axons, eventually resulting in poor synaptic signal transduction between neurons.

[0066] Unlike acute ischemic stroke, this is a chronic process which plays out over decades, but in the end the mantra “use it or lose it” becomes the dominant theme, and the entire brain network known as the default mode network degenerates, with neuronal death preferentially occurring within the Default Mode Network resulting in Alzheimer’s Disease.

[0067] Applying the methods described herein to effect push respiration in the affected tissue interrupts the damage cycle, allowing the target cells to repair themselves and slow or stop the progression of the disease. Without wishing to be bound by theory, it is believed that high frequency moderate intensity pulses likely assist with improving the function of the ATPase via push respiration even in the absence of hypoxia. Accordingly, in the case of Alzheimer’s Disease utilizing high power, high frequency pulses can be beneficial, even if it drives rapid neuronal action potentials. This is because the ATP surplus generated by push respiration allows for rapid reversal of the electron transport chain damage, such that there is long term reduction in electron leakage within the mitochondrial electron transport chain and more efficient ATP production allowing neurons and glial cells to repair. While this treatment is believed to repair cellular damage within living cells, it is not expected to result in significant neuronal or glial tissue regeneration, and as such there is no expectation that this form of treatment will be beneficial to patients who already have significant brain atrophy (moderate to severe Alzheimer’s).

[0068] Therefore, the system described herein can actively treat patients with early dementia or mild cognitive impairment, as well as the brains of all adults over the age of 50 because chronological age is the most significant risk factor for development of Alzheimer’s disease.

[0069] EQUIVALENTS

[0070] Although preferred embodiments of the invention have been described using specific terms, such description is for illustrative purposes only, and it is to be understood that changes and variations may be made without departing from the spirit or scope of the following claims.

[0071] INCORPORATION BY REFERENCE

[0072] The entire contents of all patents, published patent applications, and other references cited herein are hereby expressly incorporated herein in their entireties by reference.

Claims

CLAIMS1. A method of treating mitochondrial dysfunction, the method comprising: applying stimulation to target cells in a subject in need thereof; wherein the stimulation is configured to drive push respiration in the target cells.

2. The method of claim 1, wherein the push respiration comprises forced production of adenosine triphosphate (ATP) in the target cells.

3. The method of claim 1, wherein the stimulation includes magnetic stimulation.

4. The method of claim 3, wherein the magnetic stimulation comprises transcranial magnetic stimulation (TMS).

5. The method of any one of claims 1-4, wherein the applying stimulation step comprises varying a stimulation frequency over time.

6. The method of any one of claims 1-4, wherein the applying stimulation step comprises varying the stimulation over time with relation to a cardiac cycle of the subject.

7. The method of claim 4, wherein the TMS is administered with at least one of lower power and increased frequency.

8. The method of claim 7, wherein the lower power includes less than 260 Joules per pulse.

9. The method of claim 8, wherein: the treating includes cortical flux therapy treatment; and the lower power includes between 25 and 100 Joules per pulse.

10. The method of claim 8, wherein: the treating includes deep white matter tract treatment; and the lower power includes between 75 and 150 Joules per pulse.11 . The method of claim 7, wherein the increased frequency includes greater than 20 Hz.

12. The method of claim 7, wherein the increased frequency includes at least 30 Hz.

13. The method of claim 7, wherein the increased frequency includes at least 50 Hz.

14. The method of claim 7, wherein the TMS is administered for between 1 and 60 minutes.

15. The method of claim 7, wherein the TMS is administered for between 5 and 30 minutes.

16. The method of claim 7, wherein the TMS is administered for at least 5 minutes per hour.

17. The method of any one of claims 1-4, wherein the target cells comprise cells experiencing mitochondrial dysfunction.

18. The method of claim 17, wherein the target cells include glial cells.

19. The method of any one of claims 1-4, wherein the subject has a condition involving mitochondrial dysfunction.

20. The method of claim 19, wherein the condition includes at least one of acute acquired mitochondrial dysfunction, chronic acquired mitochondrial dysfunction, and congenital mitochondrial dysfunction.

21. The method of claim 19, wherein the mitochondrial dysfunction is from at least one of reduced oxygenation and age.

22. The method of claim 19, wherein the condition comprises ischemic stroke.

23. The method of claim 22, wherein the stimulation is administered repeatedly for 3 days following the ischemic stroke.

24. The method of claim 1, further comprising:obtaining diffusion weighted magnetic resonance imaging (MRI) data from the subject; and integrating the MRI data into the administering of the stimulation.

25. The method of claim 24, wherein the integrating of the MRI data includes interpolating three-dimensional coordinates from the MRI data into a treatment helmet.

26. The method of claim 25, wherein the integrating provides coordination between the MRI data and the stimulation.

27. A system for treating brain tissue, the system comprising: a capacitor configured to store energy from an energy source; a pulse shaper configured to release the energy from the capacitor at specific intensities and intervals; and a treatment coil configured to receive the energy from the pulse shaper and deliver the energy to the brain tissue of a subject.

28. The system of claim 27, wherein the capacitor comprises up to 160 microFarad at 1700 volts.

29. The system of claim 27, wherein the capacitor comprises up to 120 microFarad at 1700 volts.

30. The system of claim 27, wherein the system is configured to administer the method according to claim 1.

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