[go: up one dir, main page]

WO2025137265A1 - Procédés et compositions pour réduire la gliaptose : mort cellulaire neuronale induite par macroglie - Google Patents

Procédés et compositions pour réduire la gliaptose : mort cellulaire neuronale induite par macroglie Download PDF

Info

Publication number
WO2025137265A1
WO2025137265A1 PCT/US2024/061013 US2024061013W WO2025137265A1 WO 2025137265 A1 WO2025137265 A1 WO 2025137265A1 US 2024061013 W US2024061013 W US 2024061013W WO 2025137265 A1 WO2025137265 A1 WO 2025137265A1
Authority
WO
WIPO (PCT)
Prior art keywords
glial
canal
cell
neuronal
aquaporin
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/US2024/061013
Other languages
English (en)
Inventor
Ruth FABIAN-FINE
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Saint Michael's College Inc
Original Assignee
Saint Michael's College Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Saint Michael's College Inc filed Critical Saint Michael's College Inc
Publication of WO2025137265A1 publication Critical patent/WO2025137265A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia

Definitions

  • the invention in some aspects, relates to methods and compositions for reducing neuronal cell death.
  • This system is thought to facilitate waste clearance from the brain through: (a) the release of neuronal waste into interstitial spaces, (b) the existence of a convective (bulk) flow of subarachnoid cerebrospinal fluid (CFS) along para-arterial spaces first postulated by Cserr and Ostrach [H. F. Cserr and L. H. Ostrach, Experimental neurology 45, 50-60 (1974)], (c) The formation of an aquaporin4 water channel (AQP4)-mediated bulk flow of CSF into the brain parenchyma, that (d) is believed to flush cellular debris toward para-venous spaces for clearance from the brain [J. J. Iliff et al.. Science translational medicine 4, 147ral 11 (2012); J.
  • AQP4 aquaporin4 water channel
  • Impairment of this system that results in a reduced bulk flow would thus result in the failure to clear metabolites and explain waste accumulation within the brain.
  • This proposed mechanism has been challenged, in particular regarding the proposed bulk flow [ S. B. Hladky and M. A. Barrand, Fluids and Barriers of the CNS 19, 1-33 (2022) ; A. J. Smith et al., eLife 6, (2017)], and the postulated mechanism by which AQP4-containing astrocytic end feet contribute to the formation of this convective flow [N. J. Abbott et al., Acta neuropathologica 135. 387-407 (2018)].
  • Smith et al. [A. J. J.
  • a method for preventing neuronal cell death including reducing an abnormal function of a glial canal in a canalforming glial cell adjacent to the neuronal cell, wherein the reduction in the abnormal function increases likelihood of survival of the neuronal cell compared to a control likelihood of survival.
  • the control likelihood of survival is the likelihood of survival of a neuronal cell adjacent to a canal-forming glial cell in which the abnormal function is not reduced.
  • the abnormal function includes depleting the adjacent neuronal cell’s neuronal cytoplasm into the glial canal.
  • the abnormally functioning canal-forming glial cell includes one or more glial canals including structural damage.
  • the glial canal structural damage statistically significantly increases flow of cytoplasm from the neuronal cell into the structurally damaged glial canal.
  • reducing an abnormal function includes maintaining a normal function of the canal-forming glial cell.
  • the normal function of the canal-forming glial cell includes a controlled removal of neuronal waste from the adjacent neuronal cell into the glial canal of the canal-forming glial cell.
  • the neuronal waste includes one or more of lipid-based cellular waste, proteinbased cellular waste, and lipofuscin.
  • reducing the abnormal function of the canal-forming glial cell includes one or more of: increasing production of normal glial- canals by the canal-forming glial cell; maintaining a normal function of the glial-canals in the glial cell; and reducing damage to one or more glial-canals in the canal-forming glial cell.
  • the glial cell is in contact with the neuronal cell.
  • the method includes contacting the canal-forming glial cell with a composition including an aquaporin inhibitor.
  • the aquaporin inhibitor is an aquaporin 4 inhibitor.
  • the aquaporin 4 inhibitor is 2-(nicotinamide)-l,3,4- thiadiazole (TGN-020). In some embodiments, the aquaporin inhibitor is an aquaporin 7 inhibitor or is an aquaporin 9 inhibitor. In certain embodiments, the method additionally includes contacting the canal-forming glial cell with a composition including an agent that increases activity of a proteolytic enzyme. In certain embodiments, the enzyme is a caspase. In some embodiments, the caspase is a caspase 2 or a caspase 3. In some embodiments, the method includes contacting the canal-forming glial cell with a composition including an agent that reduces activity of a secretase or a BACE1.
  • the secretase is an alpha-secretase, a beta-secretase, or a gamma-s ecretas e.
  • the neuronal cell is in a subject. In certain embodiments, the subject is a mammal, optionally is a human. In some embodiments, the neuronal cell is in culture. In some embodiments, the neuronal cell is an engineered neuronal cell. In certain embodiments, the neuronal cell is in or is obtained from a subject known to have, or suspected of having, a neurodegenerative disease or condition. In some embodiments, the neurodegenerative disease or condition is Alzheimer’s disease. Parkinson’s disease. Huntington’s disease, Chronic Traumatic Encephalopathy (CTE), Amyotrophic lateral sclerosis (ALS); or Motor neuron disease. In certain embodiments, the glial cell is a macroglial cell.
  • a composition including an aquaporin inhibitor for treatment of a neurodegenerative disease or condition is provided.
  • the aquaporin inhibitor is an aquaporin 4 inhibitor, optionally wherein the aquaporin 4 inhibitor is 2-(nicotinamide)-l,3,4-thiadiazole (TGN-020).
  • the aquaporin inhibitor is an aquaporin 7 inhibitor or is an aquaporin 9 inhibitor.
  • compositions including an agent that decreases activity of a secretase for treatment of a neurodegenerative disease or condition.
  • the secretase is an alpha-secretase, a beta-secretase, or a gamma-secretase.
  • a method of reducing neurodegeneration in a subject including administering to the subject an aquaporin inhibitor in an amount effective to maintain or increase a normal function of a canal-forming glial cell in the brain of the subject.
  • the normal function of the canalforming glial cell includes a controlled removal of neuronal waste from a neuronal cell adjacent to a glial canal of the canal-forming glial cell.
  • the aquaporin inhibitor is an aquaporin 4 inhibitor, optionally wherein the aquaporin 4 inhibitor is 2- (nicotinamide)-l ,3,4-thiadiazole (TGN-020).
  • the aquaporin inhibitor is an aquaporin 7 inhibitor or is an aquaporin 9 inhibitor.
  • Figure 2A-B shows graphs indicating degeneration onset in the spider leg ganglion.
  • 3C-D are images of H&E stained tanycytes (asterisks) in the alveus form apical projections (black arrowheads) that transected into the adjacent ventricle.
  • the connectivity between adjacent somata and cell projections indicated a syncytial network.
  • Fig. 3E shows that Luxol blue stained hippocampal tanycyte processes formed varicosities (double arrowheads) that gave rise to lateral tanycyte projections (black arrow).
  • FIG. 30 shows tanycyte soma with signs of hypertrophy onset in the soma (double arrowhead).
  • Fig. 3P-S shows different focal planes through the soma of a hypertrophic tanycyte soma (double arrowhead) with attached tanycyte processes (arrow-head).
  • FIG. 4A-D provides schematic diagrams and a photomicrographic image providing a schematic summary of the proposed waste removal system in the human hippocampus and observed histopathologies.
  • Fig. 4A is a schematic diagram of myelinated aquaporin4-IR (AQP4-IR) tanycytes whose somata are located in the alveus send vast networks of tube-like processes that contain central canals into the stratum pyramidale. Tanycytes use adherence clamps to attach to surrounding cells and form intracellular receptacles that internalize catabolized neuronal w aste. This waste is transported to the ventricular lining and specialized glia canals where it may be removed from the brain via the Choroid plexus.
  • Fig. 4A is a schematic diagram of myelinated aquaporin4-IR (AQP4-IR) tanycytes whose somata are located in the alveus send vast networks of tube-like processes that contain central canals into the stratum pyramid
  • FIG. 4B is a schematic diagram of a proposed mechanism of waste uptake into tanycyte receptacles.
  • Receptacles are formed through the formation of myelin protrusions around the outer periphery of myelinated tanycytes.
  • the central, AQP4-expressing canal branches into each receptacle-forming protrusion.
  • Central canal and protrusion form a functional unit by which the functional significance of the receptacle is the filtering and catabolism of cellular waste to prevent obstruction by larger debris particles.
  • the central canal creates a convective flow- tow ard the canal through activation of AQP4.
  • the number of receptacles formed varies. Fig.
  • the invention in part, includes methods of preventing neurodegenerative diseases and conditions.
  • the instant disclosure describes studies performed to investigate waste disposal from neurons. Results show how neurons and glial cells in the invertebrate model system Cupiennius salei form a highly specialized glial-canal system by which cellular debris is cleared from neurons.
  • Central American wandering spiders are exceptionally suitable for this study as they develop similar neurodegenerative pathologies to humans that are likely linked to impaired waste clearance from the brain [R. Fabian-Fine et al., Journal of Comparative Neurology! 531, 618-638 (2023)].
  • Advantages of spider brain are their large neuronal structures, easy accessibility, superior tissue preservation, and the ability to identify initial stages of neurodegeneration based on behavioral cues of affected animals [R.
  • AQP4-IR AQP4-immunoreactive myelinated tanycytes that form waste collection ‘receptacles’ within neurons, glial cells, and extracellular spaces.
  • AQP4-IR AQP4-immunoreactive myelinated tanycytes that form waste collection ‘receptacles’ within neurons, glial cells, and extracellular spaces.
  • AQP4-IR AQP4-immunoreactive myelinated tanycytes that form waste collection ‘receptacles’ within neurons, glial cells, and extracellular spaces.
  • AQP4-IR AQP4-immunoreactive myelinated tanycytes that form waste collection ‘receptacles’ within neurons, glial cells, and extracellular spaces.
  • AD-decedents are largely obstructed by hypertrophic tanycyte receptacles, supporting a conclusion that the structural and functional failure of
  • Certain aspects of the invention comprise methods of preventing neuronal cell death in cells and subjects.
  • methods of the invention reduce an abnormal function of a canal-forming glial cell and prevent neuronal cell death.
  • glial-canal refers to a canal formed in a glial cell.
  • Spider glial canals are formed through interaction of one or more spider oligodendrocytes with tanycyte-like macroglia.
  • a mammalian canal-forming glial cell is a macroglial cell and an oligodendrocyte and is referred to herein as a tanycyte.
  • the invention in part, includes methods that maintain a normal function of a canalforming glial cell in its maintenance of a neuronal cell.
  • maintaining a normal function means reducing an abnormal function of a canal-forming glial cell.
  • Reducing an abnormal function of a canal-forming glial cell is also referred to herein as reducing an abnormal function of a glial canal in the canal-forming glial cell.
  • the term “abnormal” when used in reference to a function of a glial canal and/or a canalforming glial cell means a function that is different from a normal function of a glial canal and/or canal-forming glial cell, respectively.
  • a normal function of a glial canal and/or canal -forming glial cell comprises removing neuronal w aste from the neuronal cell into the glial canal.
  • the term “neuronal waste” as used herein means materials that are normally removed from a neuronal cell in order to maintain the health of the neuronal cell.
  • Non-limiting examples of materials that may be included in neuronal waste are lipid- based cellular waste, protein-based cellular waste, and lipofuscin.
  • '‘normal” as used herein in reference to removal of neuronal waste from a neuronal cell means removal of the neuronal cell waste in a controlled manner. Controlled removal of waste is important for maintaining the health of the neuronal cell and removal of such waste material is a normal function of glial canals and/or canal-forming glial cells. In contrast, an abnormal function of a glial canal and/or canal-forming glial cell comprises uncontrolled removal of cytoplasm from a neuronal cell adjacent to the canal-forming glial cell.
  • abnormal function of a glial canal and/or canal-forming glial cell essentially evacuates cytoplasm from the neuronal cell in an uncontrolled manner, resulting in excess removal and/or depletion of the cytoplasm of the neuronal cell.
  • An abnormally functioning glial canal can deplete the cytoplasm of a neuronal cell physically adjacent to the canal-forming cell comprising the glial-canal, resulting in death of both the neuronal cell and additional cells, such as surrounding oligodendroglia.
  • an abnormal depletion of cytoplasm of a neuronal cell may result in abnormal removal of cytoplasm from a neuronal cell adjacent to a canal-forming glial cell results in death of the neuronal cell, the canal-forming glial cell, and additional oligodendroglia.
  • a canal-forming glial cell is an oligodendroglial cell.
  • Mammalian canal-forming glial cells are tanycytes.
  • a chain-reaction response may occur resulting not only in the death of the neuronal cell and the canal-forming glial cell, but additional cells such as oligodendroglial cells may also die as a result of the release of enzymes and other materials by the neuronal cell, canal-forming glial cell, and/or other negatively impacted cells upon their death.
  • An abnormal function of one or more glial canals and/or canal-forming glial cells may result from structural damage to the one or more glial canals and/or canal-forming glial cells, respectively.
  • the presence of structural damage of a glial canal statistically significantly increases the level of flow of cytoplasm from a neuronal cell, such that a neuronal cell adjacent to a canal-forming glial cell comprising one or more structurally damaged glial canals experiences uncontrolled and lethal outflow 7 of its cytoplasm.
  • the normal function of removing waste from a neuronal cell includes removing lipid-based cellular w aste, protein-based cellular waste, and/or lipofuscin cellular waste, which may be removed in conjunction an amount of neuronal cytoplasm. It will be understood that the amount of neuronal cytoplasm removed in the context of normal waste removal from a neuronal cell into a normally functioning glial canal is an amount that does not negatively impact the neuronal cell.
  • an amount of neuronal cell cytoplasm removed into an abnormally functioning glial canal may be at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 120%, 140%, 160%, 180%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, or more than the amount of neuronal cell cytoplasm removed into a normally functioning glial canal.
  • a mammalian neuronal cell is contacted by one or more tanycytes that form varicose processes into the neuronal cell for removal of neuronal waste. Canals in the processes project membrane-cistemae into the neuronal cytoplasm permitting controlled removal of neuronal waste from the interior of the neuronal cell into the tanycyte.
  • a mammalian neuronal cell is contacted by one or more tanycytes that form varicose processes into the neuronal cell for removal of neuronal waste.
  • methods of the invention comprise reducing an abnormal function of a canal-forming glial cell, which, as indicated herein, reduces an abnormal function of one or more glial canals in the canal -forming glial cell.
  • methods of the invention include maintaining a normal function of the canal-forming glial cell, which as indicated herein, means maintaining a normal function of one or more glial canals of the canal-forming glial cell.
  • Maintaining a normal function of a canal-forming glial cell may include one or more of (a) increasing production of normal glial canals by the canalforming glial cell, and (b) reducing damage, for example the occurrence of structural damage, to one or more glial-canals in the canal-forming glial cell.
  • an abnormal function of a glial canal can be reduced by contacting a canal-forming glial cell with a composition comprising an aquaporin inhibitor agent.
  • Methods of the invention comprise contacting a canal-forming glial cell with a composition comprising one or more aquaporin inhibitor agents, non-limiting examples of which are aquaporin 4 inhibitor agents, aquaporin 7 inhibitor agents, and aquaporin 9 inhibitor agents.
  • a canal-forming glial cell is contacted with a composition comprising one or more aquaporin inhibitors in an amount effective to reduce an abnormal function of a glial canal in the contacted canalforming glial cell.
  • reduce as used herein in reference to a change in an abnormal function of a glial canal and/or canal-forming glial cell means decreasing the abnormal function, which may include fully preventing, partially preventing, inhibiting, and/or eliminating the abnormal function.
  • the composition comprising the aquaporin inhibitor comprises an aquaporin 4 inhibitor agent.
  • Aquaporin 4 inhibitor agents are 2- (nicotinamide)-l,3,4-thiadiazole (TGN-020), IMD 0354, zinc [see Yukutake Y, et al., Biochemistry. 2009 Dec 29;48(51): 12059-61], and an anti-aquaporin 4 antibody or functional fragment thereof.
  • a canal-forming glial cell is contacted with a composition comprising an aquaporin 7 inhibitor agent.
  • Non-limiting examples of Aquaporin 7 inhibitor agents are. Z433927330 (for example: Cat.
  • HY- 126074 MCE Med Chem Express
  • monoacetin monobutyrin and diacetin
  • an anti-aquaporin 7 antibody or functional fragment thereof an anti-aquaporin 7 antibody or functional fragment thereof.
  • a canalforming glial cell is contacted with a composition comprising an aquaporin 9 inhibitor agent.
  • Aquaporin 9 inhibitor agents are RG100204 (see for example, U.S. Patent Publication US20190127360) and an anti-aquaporin 9 antibody or functional fragment thereof. (Each of the above publications is incorporated by reference herein in its entirety .)
  • a combination of aquaporin inhibitor agents may be selected and included in a composition that contacts a canal-forming glial cell in a method of the invention.
  • a canal-forming glial cell may be contacted with a composition comprising one, two, or more of aquaporin inhibitor agents, which may be independently selected from an aquaporin 4 inhibitor agent, an aquaporin 7 inhibitor agent, and an aquaporin 9 inhibitor agent.
  • the term “independently selected” used in reference to aquaporin inhibitor agents means each agent may be chosen to be administered for contacting a canal-forming glial cell independent of the selection of one or more other aquaporin inhibitor agents.
  • a subject may be administered three aquaporin inhibitor agents and the selected agents are: TGN-20, Z433927330, and an anti-aquaporin 4 antibody.
  • a composition comprising one or more aquaporin inhibitor agents comprise one or more components in addition to the aquaporin inhibitor agent(s), non-limiting examples of additional components that may be included in the composition are detectable labels, earners, delivery agents, etc.
  • contacting a neuronal cell or cells with a composition comprising an aquaporin inhibitor agent reduces an abnormal function and increases the likelihood of survival of one or more neuronal cells normally maintained by the canal-forming glial cell contacted with the composition, as compared to a control likelihood of survival.
  • a control likelihood of survival is a likelihood of survival of one or more neuronal cells adjacent to a canal -forming glial cell that is not contacted with the composition.
  • Certain methods of the invention include a treatment regimen comprising administering to a subject identified as having, or at risk of having, a neurodegenerative disease or condition, one or more aquaporin inhibitor agents.
  • Certain embodiments of methods of the invention may include (1) identifying a subject who is at risk of having neuronal degeneration or a subject who has neuronal degeneration, (2) selecting a therapeutic regimen with which to treat the subject identified in step (1); and (3) administering the selected therapeutic regimen to the subject identified as in need of the selected therapeutic regimen.
  • a reduced abnormal function of a glial canal of a canal-forming glial cell that normally functions to maintain a neuronal cell can be treated by contacting the canal-forming glial cell with one or more aquaporin inhibitor agents.
  • Certain embodiments of methods of the invention can be used to prevent and/or treat a subject by administering a composition comprising one or more aquaporin inhibitor agents to the subject in an amount effective to reduce an abnormal glial-canal function and prevent and/or reduce the severity of neuronal death and neuronal degeneration in the subject.
  • Certain methods of the invention include administering composition(s) comprising one or more aquaporin inhibitor agents to a subject identified as having a neuronal degenerative disease or condition or administering a composition comprising one or more aquaporin inhibitor agents prophy lactically to a subject identified as at risk of having a neuronal degenerative disease or condition.
  • Methods that can be used to identify a subject as at risk of having or as having a neuronal degenerative disease or condition are known in the art, and may include but are not limited to imaging methods, behavioral assessment, genetic analysis, etc.
  • an abnormal function of a glial canal may be reduced by contacting a canal-forming glial cell with a composition comprising an agent that increases activity of an enzyme, a non-limiting example of which is a caspase.
  • an enzyme such as a caspase
  • increasing activity of an enzyme, such as a caspase may assist in the degradation of w aste products that are then removed into canal-forming glial cells.
  • methods of the invention comprise contacting a canal-forming glial cell with a composition comprising one or more agents that increase activity of a caspase, non-limiting examples of w hich caspase 2 and caspase 3.
  • a canal-forming glial cell is contacted with a composition comprising one or more agents that increase activity’ of a caspase 2.
  • a canal-forming glial cell is contacted with a composition comprising one or more agents that increase activity- of a caspase 3 in an amount effective to reduce an abnormal function of a glial canal in the contacted canal-forming glial cell.
  • the term “reduce” as used herein in reference to a change in an abnormal function of a glial canal and/or canal-forming glial cell means decreasing the abnormal function, which may include fully preventing, partially preventing, inhibiting, and/or eliminating the abnormal function.
  • an abnormal function is the failure of the glial canal and/or canal- formal glial cells to remove waste.
  • methods of treating a neurodegenerative disease or condition comprise increasing activity of an enzyme in a subject believed to have or to be at risk of having the neurodegenerative disease or condition.
  • Caspase enzymes are non-limiting example of enzymes that methods of the invention may be used to increase to treat a neurodegenerative disease or condition.
  • Caspase 2 and caspase 3 are non-limiting examples of caspase enzymes whose activity’ may be increased in a subject using a method of the invention.
  • activity of caspase 2 and/or caspase 3 is increased in an amount effective to treat a neurodegenerative disease or condition in a subject.
  • Some embodiments of methods of the invention comprise increasing a level of activity of a caspase 2 or caspase 3 enzyme.
  • a method of the invention increases activity of an enzyme in neuronal tissue of a subject by one or more of: increasing expression of the enzyme, reducing loss of the enzyme, increasing enzymatic functioning of the enzyme, and other means that results in a higher level of activity of the enzy e in the neuronal tissue of the subject.
  • one or more agents that increase a level of activity of an enzy me such as but not limited to a caspase, are administered to a subject.
  • a level of activity of an enzyme may be determined before and/or after a treatment of the invention is administered to a subject.
  • a level of activity of an enzyme such as but not limited to a caspase in a subject being treated or to be treated with a method of the invention, may be compared to a control level of activity of the enzyme to identify efficacy of the treatment of the invention. In such instances, whether or not the treatment results in an increase in enzyme activity in the subject can be determined using such comparisons.
  • a control level of activity is a level of activity in a subject or subjects that do not have the degenerative disease or condition that is present in a subject or suspected to be present in the subject.
  • a control level may be a level of enzyme activity in the subject prior to administering a treatment of the invention.
  • the control level may be compared with a level of activity' of the enzyme following a treatment of the subject with a method of the invention as a determination of the efficacy of the treatment against the degenerative disease or condition in the subject.
  • determining a level of activity of an enzyme, such as but not limited to a caspase enzyme in a subject may include but is not limited to one of more of imaging studies, symptom assessment, and other diagnostic methods with which to determine the status of the neurodegenerative disease or condition.
  • a treatment method of the invention may result in a higher level of activity of an enzyme such as, but not limited to, a caspase following the treatment compared to a level of activity of the enzyme prior to the treatment. It will be understood that a “normal” level of activity may be used as a control level.
  • an activity level of an enzyme such as a caspase in a neuronal tissue of the subject may be at a level that is below a “normal” activity level and increasing activity in that subject may be an increase of activity in that subject to a level that is closer to, equal to, or greater than a normal level of activity of the enzyme.
  • a subject in need of treatment with a method of the invention has an essentially normal level of activity of an enzyme such as, but not limited to, a caspase in their neuronal tissues and increasing the enzyme activity in that subject may be an increase to a level greater than a normal level of activity.
  • the subject may be treated by providing a higher-than-normal level of activity of the enzyme, thereby treating the neurodegenerative disease or condition in the subject.
  • methods of treating a neurodegenerative disease or condition comprise decreasing activity of an enzyme in a subject believed to have or to be at risk of having the neurodegenerative disease or condition.
  • alpha-, beta- and/or gamma- secretases may promote the growth of the tanycytes into neurons and the proliferation of receptacles. Results indicate that secretases may promote outgrow th and proliferation of blood vessels through interaction with amyloid precursor protein (APP) that is cleaved by an enzy me called "beta-site APP cleaving enzyme 1” (BACE1).
  • APP amyloid precursor protein
  • Some embodiments of methods of the invention comprise reducing activity of a secretase and/or a BACE1 enzyme reduces proliferation of excessive receptacles in the human AD brain.
  • BACE1 and secretase enzymes are non-limiting examples of enzy mes that methods of the invention may be used to decrease to treat a neurodegenerative disease or condition.
  • Alpa-, beta-, and/or gamma-secretase enzymes are non-limiting examples of secretase enzymes whose activity may be decreased in a subject using a method of the invention.
  • activity’ of one or more of alpha-secretase, beta- secretase, and gamma-secretase is decreased in an amount effective to treat a neurodegenerative disease or condition in a subject.
  • Some embodiments of methods of the invention comprise decreasing a level of activity of one or more of alpha-secretase, beta- secretase, and gamma-secretase.
  • a method of the invention decreases activity of an enzyme in neuronal tissue of a subject by' one or more of: decreasing expression of the enzyme, increasing elimination (for example increasing removal or degradation) of the enzy me, decreasing enzymatic functioning of the enzyme, and other means that results in a lower level of activity of the enzyme in the neuronal tissue of the subject.
  • one or more agents that decrease a level of activity of a secretase such as but not limited to an anti- secretase antibody, a secretase-silencing RNA, are administered to a subject.
  • a level of activity of an enzyme may be determined before and/or after a treatment of the invention is administered to a subject. Comparing a level of activity of an enzyme, such as but not limited to a secretase or BACE1, in a subject to a control level of activity of the enzyme may be used to identify efficacy of a treatment of the invention, by determining whether or not the treatment results in a decrease in secretase activity or BACE1 activity in the subject.
  • a control level of activity is a level of activity in a subject or subjects that do not have the degenerative disease or condition that is present in a subject or suspected to be present in the subject being treated or to be treated with a method of the invention.
  • a control level may be a level of enzy me activity in the subject prior to administering a treatment of the invention.
  • the control level maybe compared with a level of activity of the secretase enzyme and/or BACE1 activity following a treatment of the subject with a method of the invention as a determination of the efficacy of the treatment against the degenerative disease or condition in the subject.
  • determining a level of activity of an enzyme may include but is not limited to one of more of imaging studies, symptom assessment, and other diagnostic methods with which to determine the status of the neurodegenerative disease or condition.
  • a treatment method of the invention may result in a lower level of activity 7 of an enzy me such as, but not limited to, a secretase and/or BACE1 following the treatment compared to a level of activity of the enzyme prior to the treatment. It will be understood that a “normal” level of activity 7 may be used as a control level.
  • an activity' level of an enzyme such as a secretase or BACE1 in a neuronal tissue of the subject may be at a level that is above a “normal”’ activity- level and decreasing activity in that subject may be a decrease of activity in that subject to a level that is closer to, equal to, or lower than a normal level of activity of the enzyme.
  • a subject in need of treatment with a method of the invention has an essentially normal level of activity of an enzy me such as, but not limited to, a secretase and/or BACE1, in their neuronal tissues and decreasing the enzyme activity- in that subject may be a decrease to a level less than a normal level of activity.
  • the subject may be treated by providing a lower-than-normal level of activity of the enzyme, thereby treating the neurodegenerative disease or condition in the subject.
  • the term “at least a portion” used in reference to activity means at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%. 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%. 22%, 23%. 24%. 25%. 26%. 27%. 28%. 29%. 30%. 31%. 32%.
  • Efficacy of a method of the invention to reduce a subject’s risk may be determined by comparing results of administering an aquaporin inhibitor agent to a subject with control results.
  • an aquaporin inhibitor agent administered to a subject reduces the subject’s risk of developing a neurodegenerative disease or condition compared to a control risk of developing the neurodegenerative disease or condition, wherein the control risk is a risk of a subject in essentially identical circumstances developing the neurodegenerative disease or condition in the absence of the administered aquaporin inhibitor agent.
  • Efficacy of a method of the invention to reduce a subject’s risk may be determined by comparing results of administering an agent that decreases activity of an enzyme, such as but not limited to a secretase and/or BACE1 to a subject with control results.
  • an agent that decreases activity of a secretase and/or BACE1 is administered to a subject and reduces the subject’s risk of developing a neurodegenerative disease or condition compared to a control risk of developing the neurodegenerative disease or condition, wherein the control risk is a risk of a subject in essentially identical circumstances developing the neurodegenerative disease or condition in the absence of the administered agent that decreases activity of a secretase and/or BACE1.
  • Certain embodiments of methods of the invention comprise administering an agent that reduces an abnormal function of a glial canal in a canal-forming glial cell in the subject (a non-limiting example of which is an aquaporin inhibitor agent) in an amount effective to treat a neurodegenerative disease or condition.
  • Some embodiments of methods of the invention comprise administering an agent that increases activity of an enzyme, such as but not limited to a caspase, thereby reducing an abnormal function of a glial canal in a canalforming glial cell in the subject in an amount effective to treat a neurodegenerative disease or condition in the subject.
  • Some embodiments of methods of the invention comprise administering an agent that decreases activity of an enzyme, such as but not limited to a secretase and/or BACE1, thereby reducing an abnormal function of a glial canal in a canalforming glial cell in the subject in an amount effective to treat a neurodegenerative disease or condition in the subject.
  • An effective amount is a dosage of the agent sufficient to provide a medically desirable result.
  • pharmacological agents of the invention are used to treat or prevent neurodegenerative diseases or conditions, that is, in some embodiments they may be used to treat an existing neurodegenerative disease or condition in a subject, and they may also be administered prophy lactically to a subject at risk of developing a neurodegenerative disease or condition.
  • An effective amount is that amount that can lower a risk of, slow or perhaps prevent altogether the development of a neurodegenerative disease or condition in a subject.
  • Factors involved in determining an effective amount are well known to those of ordinary skill in the art and can be addressed with no more than routine experimentation. It is generally preferred that a maximum dose of a pharmacological agent of the invention be used, that is, the highest safe dose according to sound medical judgment. It will be understood by those of ordinary skill in the art however, that a subject (also referred to herein as a patient) may insist upon a lower dose or tolerable dose for medical reasons, psychological reasons or for virtually any other reasons.
  • the therapeutically effective amount of a pharmacological agent of the invention is that amount effective to treat the condition, such as a neurodegenerative disease or condition.
  • the desired response is inhibiting the progression of the neurodegenerative disease or condition and/or reducing the severity and/or the level of the neurodegenerative disease or condition. This may involve only slowing the progression of the neurodegenerative disease or condition temporarily, although it may include halting the progression of the neurodegenerative disease or condition permanently. This can be monitored by routine diagnostic methods known to those of ordinary skill in the art.
  • a desired response to a method of the invention to treat a neurodegenerative disease or condition may in some embodiments, be preventing the onset of the neurodegenerative disease or condition.
  • Certain embodiments of methods of the invention include administering an agent (non-limiting examples of which are an aquaporin inhibitor agent, an agent that increases a level of caspase activity, and an agent that decreases a level of secretase activity and/or BACE1 activity ) that reduces an abnormal function of a glial canal in a canal-forming glial cell in the subject, wherein the agent is administered in an amount effective to reduce the subject's risk of developing a neurodegenerative disease or condition, and/or to reduce the severity of a neurodegenerative disease or condition present in the subject.
  • a therapeutically effective amount refers to that amount of the agent being administered to a subject that is sufficient to prevent progression of a neurodegenerative disease or condition.
  • Administration of the agent in an amount effective to reduce an abnormal function of a glial canal in a canal-forming glial cell in the subject may reduce the risk of a subject developing the neurodegenerative disease or condition by at least 1%, 2%, 3%, 4%, 5%. 6%, 7%, 8%.
  • administering an effective amount of an aquaporin inhibitor agent to the subject may reduce the subject’s 20% risk down to less than 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% risk, or to 0% risk.
  • administering an effective amount of an agent that reduces an abnormal function of a glial canal in a canal-forming glial cell in the subject increases the likelihood of survival of neuronal cells in the subject compared to a control likelihood of survival.
  • a control likelihood of survival of neuronal cells is a likelihood of survival of the neuronal cells in a subject in the absence of the administration of the effective amount of the agent.
  • Administration of an effective amount of the agent to a subject in need of such treatment can increase the likelihood of survival neuronal cells in the subject to least 1, 2, 3, 4, 5. 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 100 times higher than the control likelihood of survival of neuronal cells in the subject.
  • Another way of expressing a change in likelihood of survival of neuronal cells is in reduction in the percent likelihood of neuronal cell death due to the neurodegenerative disease or condition.
  • the subject may have a risk of neuronal cell death that is up to 1%. 2%, 3%, 4%, 5%, 6%. 7%, 8%, 9%. 10%.
  • a control risk of neuronal cell death from a neurodegenerative disease or condition is 80%
  • a therapeutic regimen comprising administration of one or more agents that reduce an abnormal function of a glial canal in a canal-forming glial cell in the subject (non-limiting examples of which are an aquaporin inhibitor agent and agent, an agent that increases a level of caspase activity, and an agent that decreases a level of secretase activity and/or BACE1 activity) to a subject determined to be in need such treatment may include administration of one or more agents once, or multiple times.
  • Multiple administrations of an agent that reduces an abnormal function of a glial canal in a canal-forming glial cell in the subject means the agent is administered to a subject 2, 3, 4, 5, 6, 7, 8, 9, 10, or more times.
  • an agent that reduces an abnormal function of a glial canal in a canal-forming glial cell in the subject may be done in combination with additional treatments for a neurodegenerative disease or condition.
  • a therapeutic regimen (also referred to herein as a treatment) may be selected for a subject based at least in part on the identification that the subject is at risk of having or that the subject has neuronal degeneration.
  • selection of a treatment may be based, at least in part, on the severity of a neuronal degenerative disease or condition in a subject.
  • a selected treatment may include administering to the subject an effective amount of one or more agents that reduce an abnormal function of a glial canal in a canal-forming glial cell adjacent to a neuronal cell.
  • the administered agent is an aquaporin inhibitor agent.
  • Certain embodiments of methods of the invention include administering an agent that reduces an abnormal function of a glial canal and administering one or more additional treatments appropriate for the specific neurodegenerative disease or condition in the subject.
  • an additional treatment for a subject identified as having or at risk of having a neurodegenerative disease or condition includes one or more of physical therapy, surgery, administration of one or more additional therapeutic agents, dietary modification, etc. Upon a determination of the presence of, or a risk of.
  • a practitioner will, without undue experimentation, be aware of and able to select one or more treatments that may be administered to a subject in addition to the administration of an agent that reduces an abnormal function of a glial canal, non-limiting examples of which are an aquaporin inhibitor agent, an agent that increases a level of caspase activity, and an agent that decreases a level of secretase activity and/or BACE1 activity.
  • an agent that reduces an abnormal function of a glial canal non-limiting examples of which are an aquaporin inhibitor agent, an agent that increases a level of caspase activity, and an agent that decreases a level of secretase activity and/or BACE1 activity.
  • Neurodegenerative disease 7 and “neurodegenerative” are used interchangeably herein in reference to diseases and conditions.
  • diseases or conditions include, but are not limited to Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, Amyotrophic lateral sclerosis (ALS); Chronic Traumatic Encephalopathy (CTE), and Motor Neuron Disease.
  • ALS Amyotrophic lateral sclerosis
  • CTE Chronic Traumatic Encephalopathy
  • a neurodegenerative disease or condition is a disease or condition in which there is degeneration and death of neuronal cells.
  • Some embodiments of methods of the invention may include selecting a therapeutic regimen for a subject, wherein the therapeutic regimen comprises administering to the subject an agent that reduces an abnormal function of a glial canal in a canal -forming glial cell in the subject.
  • an agent that reduces an abnormal function of a glial canal in a canal -forming glial cell in the subject.
  • a non-limiting example of such an agent is an aquaporin inhibitor agent.
  • a treatment method of the invention may also include one or more additional therapeutic actions or administered medicaments, depending on the specific neurodegenerative disease or condition, the severity of the neurodegenerative disease or condition, or other factors of which a practitioner will be aware as factors for consideration in selecting a treatment.
  • Methods of the invention include producing in a subject in need of such treatment a therapeutic effect against neuronal cell death and/or a neurodegenerative disease or condition.
  • therapeutic effect as used herein in reference to an agent that reduces an abnormal function of a glial canal in a canal-forming glial cell means a clinically beneficial effect of the agent against neuronal cell death and/or a neurodegenerative disease or condition when it is administered to a subject in need of such treatment.
  • a therapeutic effect of an agent that reduces an abnormal function of a glial canal in a canal-forming glial cell in the subject can be determined, for example, by detecting one or more physiological effects of the treatment, such as the decrease or lack of symptoms of the neurodegenerative disease or condition following administration of the treatment.
  • Additional means of monitoring and assessing a neurodegenerative disease or condition in a subject, and ways to assess and determine one or more of a level, severity, change in severity, etc. of a neurodegenerative disease or condition in subject are known in the art and can be used to assess the neurodegenerative condition in a subject following a treatment method of the invention.
  • Non-limiting examples of physiological symptoms of neurodegenerative diseases and conditions that may be assessed in certain embodiments of methods of the invention are provided elsewhere herein and will be known in the art.
  • Methods and compositions of the invention may be used to treat a neurodegenerative disease or condition.
  • the terms “treat”, “treated”, or “treating” when used in relation to a neurodegenerative disease or condition may refer to a prophylactic treatment that decreases the likelihood or risk of a subject developing the neurodegenerative disease or condition, and may be used to refer to a treatment after a subject has developed a neurodegenerative disease or condition in order to eliminate or ameliorate the neurodegenerative disease or condition, prevent the neurodegenerative disease or condition from becoming more advanced or severe, and/or to slow the progression of the neurodegenerative disease or condition compared to the progression of the neurodegenerative disease or condition in the absence of a therapeutic method of the invention.
  • a subject may be a vertebrate animal including but not limited to a human, mouse, rat, guinea pig, rabbit, cow, dog, cat, horse, goat, and non -human primate, e.g., monkey.
  • a subject may be a mammal.
  • a subject is any human or non-human recipient of one or more of an aquaporin-inhibitor agents, a caspase-activity- increasing agent, a secretase-activity reducing agent, a BACE1 -activity reducing agent, a composition, or a pharmaceutical composition of the invention as described herein.
  • a subject may be a domesticated animal, a wild animal, or an agricultural animal.
  • a subject is a human.
  • a subject has or is at risk of having a neurodegenerative disease or condition and is in need of a treatment of the invention.
  • a subject does not have epilepsy or a seizure disorder.
  • a subject does not have cytotoxic brain swelling.
  • a subject does not have fluid- induced brain swelling.
  • a subject does not have brain oedema.
  • a subject does not have a brain tumor.
  • a neuronal cell may be a cell obtained from a cell sample, tissue sample, blood sample, etc.
  • a method of the invention may include contacting a cell with a composition of the invention in vitro, ex vivo, or in vivo.
  • a method of the invention comprises contacting a canal-forming neuronal cell that is in culture.
  • a canal-forming glial cell that is in a subject.
  • Certain embodiments of methods of the invention include ex vivo treatment methods.
  • a neurodegenerative disease or condition in a subject can be detected using an art- known method, certain of which are described elsewhere herein.
  • Methods that may be used to detect a neurodegenerative disease or condition include, but are not limited to identifying the presence of one or more physiological characteristics or symptoms of the neurodegenerative disease or condition in the cell or subject, assessing genetic characteristics of a cell or subject, histological and/or imaging methods applied to a cell and/or a subject, etc.
  • Characteristics of a neurodegenerative disease or condition detected in a subject can be compared to control values of the characteristics of the neurodegenerative disease or condition.
  • a control value may be a predetermined value, which can take a variety of forms. It can be a single cut-off value, such as a median or mean.
  • comparative groups such as in groups of individuals having the neurodegenerative disease or condition, groups of individuals who have been administered a treatment for the neurodegenerative disease or condition, groups of individuals who have not been administered a treatment for the neurodegenerative disease or condition, etc.
  • Another example of comparative groups may be groups of subjects having one or more symptoms of or a diagnosis of the neurodegenerative disease or condition and groups of subjects without the one or more symptoms of or a diagnosis of the neurodegenerative disease or condition.
  • a predetermined value will depend upon the particular population selected. Accordingly, the predetermined value selected may take into account a category in which an individual falls. Non-limiting examples of categories are a subject’s age, a subject’s genetic background, etc. Appropriate categories can be selected and utilized with no more than routine experimentation by those of ordinary skill in the art.
  • a comparison of a treated versus a control may include comparing disease severity differences between a treated subject and a selected control.
  • severity of symptoms or physiological effects of a neurodegenerative disease or condition in a subject treated with a method of the invention may be determined to be less relative to a selected control, with the comparison indicating up to a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%,
  • a level of severity' of a treated subject’s neurodegenerative disease or condition is less than 100% of a control severity level of the neurodegenerative disease or condition.
  • the severity of one or physiological symptoms of the neurodegenerative disease or condition in a subject treated according to a method of the invention is less than or equal to 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%. 86%. 85%. 84%. 83%. 82%. 81%. 80%. 79%.
  • controls may be, in addition to predetermined values, samples of materials tested in parallel with the experimental materials. Examples include samples from control populations or control samples generated through manufacture to be tested in parallel with the experimental samples; and a control may be a sample from a subject prior to, during, or after a treatment with an embodiment of a method or composition of the invention. Thus, one or more characteristics determined for a subj ect having a neurodegenerative disease or condition may be used as “control” values for those characteristics in that subject at a later time.
  • the pharmacological agents used in the methods of the invention are preferably sterile and contain an effective amount of an aquaporin inhibitor agent, an effective amount of an agent that increases a level of caspase activity, or an agent that decreases a level of secretase activity and/or decreases a level of BACE1 activity, for producing the desired response in a unit of weight or volume suitable for administration to a subject.
  • an agent that reduces an abnormal function of a glial canal in a canal-forming glial cell in the subject is delivered in a composition formulated to cross into the brain, e.g., formulated to cross the blood-brain barrier.
  • a composition formulated to cross into the brain e.g., formulated to cross the blood-brain barrier.
  • Doses of pharmacological agents administered to a subject can be chosen in accordance with different parameters, in particular in accordance with the mode of administration used and the state of the subject. Other factors include the desired period of treatment.
  • a pharmacological agent may be adjusted by the individual healthcare provider or veterinarian, particularly in the event of any complication.
  • a therapeutically effective amount typically varies from 0.01 mg/kg to about 1000 mg/kg, from about 0.1 mg/kg to about 200 mg/kg, or from about 0.2 mg/kg to about 20 mg/kg, in one or more dose administrations daily, for one or more days.
  • An agent that reduces an abnormal function of a glial canal in a canal -forming glial cell in the subject may also be referred to herein as a pharmacological agent.
  • the amount of a treatment of the invention administered to a subject may be varied for example by increasing or decreasing the amount of one or more agents administered to the subject to reduce an abnormal function of a glial canal in a canal -forming glial cell in the subject.
  • Changes in a treatment of the invention may include one or more of changing the therapeutic composition administered, changing the route of administration, changing the dosage timing and so on.
  • An effective amount of a composition of the invention will vary with the particular neurodegenerative disease or condition being treated, the age and physical condition of the subject being treated, the severity of the neurodegenerative disease or condition, the duration of the treatment, the specific route of administration, and other art- known factors within the know ledge and expertise of a health practitioner.
  • Treatment compositions and treatment agents of the invention are also referred to herein as pharmaceutical agents, pharmaceutical compounds, and pharmaceutical compositions of the invention.
  • Treatment compositions of the invention may be administered to mammals other than humans, e.g., for testing purposes or veterinary therapeutic purposes, and such administrations may be carried out under substantially the same conditions as described herein. It will be understood that methods and compositions of the invention are applicable to both human and animal diseases. Thus, this invention is intended to be used in husbandry and veterinary medicine as well as in human therapeutics.
  • the pharmaceutical preparations of the invention When administered, the pharmaceutical preparations of the invention are applied in pharmaceutically acceptable amounts and in pharmaceutically acceptable compositions.
  • ⁇ pharmaceutically acceptable means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredients.
  • Such preparations may routinely contain salts, buffering agents, preservatives, compatible carriers, and optionally other therapeutic agents.
  • the salts When used in medicine, the salts should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts may conveniently be used to prepare pharmaceutically acceptable salts thereof and are not excluded from the scope of the invention.
  • Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, maleic, acetic, salicylic, citric, formic, malonic, succinic, and the like.
  • pharmaceutically acceptable salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, potassium or calcium salts.
  • a pharmacological agent or composition of the invention may be combined, if desired, with a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier means one or more compatible solid or liquid fillers, diluents, or encapsulating substances which are suitable for administration into a human.
  • carrier denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate the application.
  • the components of the pharmaceutical compositions also are capable of being co-mingled with the pharmacological agents of the invention, and with each other, in a manner such that there is no interaction which would substantially impair the desired pharmaceutical efficacy.
  • the pharmaceutical compositions may contain suitable buffering agents, non-limiting examples of which are acetate, phosphate, citrate, glycine, borate, carbonate, bicarbonate, hydroxide (and other bases) and pharmaceutically acceptable salts of the foregoing compounds.
  • suitable buffering agents non-limiting examples of which are acetate, phosphate, citrate, glycine, borate, carbonate, bicarbonate, hydroxide (and other bases) and pharmaceutically acceptable salts of the foregoing compounds.
  • suitable preservatives such as: benzalkonium chloride; chlorobutanol; parabens and thimerosal.
  • compositions suitable for oral administration may be presented as discrete units, such as capsules, tablets, pills, lozenges, each containing a predetermined amount of the active compound (e.g., an aquaporin inhibitor).
  • Other compositions include suspensions in aqueous liquids or non-aqueous liquids such as a syrup, elixir, an emulsion, or a gel.
  • Pharmaceutical preparations for oral use can be obtained as solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores.
  • Non-limiting examples of suitable excipients include fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP).
  • disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
  • the oral formulations may also be formulated in saline or buffers, i.e., EDTA for neutralizing internal acid conditions or may be administered without any carriers.
  • the location of release may be the stomach, the small intestine (the duodenum, the jejunum, or the ileum), or the large intestine.
  • the stomach the small intestine (the duodenum, the jejunum, or the ileum), or the large intestine.
  • One skilled in the art has available formulations which will not dissolve in the stomach yet will release the material in the duodenum or elsewhere in the intestine.
  • the release will avoid the deleterious effects of the stomach environment, either by protection of the aquaporin inhibitor agent or by release of the biologically active material beyond the stomach environment, such as in the intestine.
  • the treatment agents of the invention when it is desirable to deliver them systemically, may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion.
  • Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative.
  • the compositions of the invention may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents.
  • compositions for parenteral administration include aqueous solutions of the active compounds in water-soluble form.
  • Suspensions of the active compounds may be prepared as appropriate oily injection suspensions.
  • Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes.
  • Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran.
  • the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
  • the active compounds may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
  • the particle may include, in addition to the pharmacological agent(s), any of those materials routinely used in the art of pharmacy and medicine, including, but not limited to, erodible, nonerodible, biodegradable, or nonbiodegradable material or combinations thereof.
  • the particles may be microcapsules which contain the aquaporin inhibitor agent in a solution or in a semi-solid state.
  • the particles may be of virtually any shape.
  • Both non-biodegradable and biodegradable polymeric materials can be used in the manufacture of particles for delivering the pharmacological agent(s).
  • Such polymers may be natural or synthetic polymers. The polymer is selected based on the period of time over which release is desired.
  • Bioadhesive polymers of particular interest include bioerodible hydrogels described by H. S. Sawhney, C. P. Pathak and J. A. Hubell in Macromolecules, (1993) 26:581-587, the teachings of which are incorporated herein.
  • polyhyaluronic acids include polyhyaluronic acids, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylates), poly(ethyl methacrylates), poly(butylmethacrylate), poly(isobutyl methacrylate), poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate). poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acry late), and poly(octadecyl acrylate).
  • a pharmacological agent of the invention may be contained in controlled-release systems.
  • controlled release 7 is intended to refer to any drug-containing formulation in which the manner and profile of drug release from the formulation are controlled. This refers to immediate as well as non-immediate release formulations, with non-immediate release formulations including but not limited to sustained-release and delayed-release formulations.
  • sustained release also referred to as “extended release” is used in its conventional sense to refer to a drug formulation that provides for gradual release of a drug over an extended period of time, and that may, although not necessarily, results in substantially constant blood levels and/or tissue levels of a drug over an extended time period.
  • delayed release is used in its conventional sense to refer to a drug formulation in which there is a time delay between administration of the formulation and the release of the drug therefrom. “Delayed release” may or may not involve gradual release of drug over an extended period of time, and thus may or may not be “sustained release.”
  • kits can include one or more pharmaceutical preparation vial, a pharmaceutical preparation diluent vial, and an aquaporin inhibitor agent.
  • a vial containing the diluent for the pharmaceutical preparation is optional.
  • a diluent vial may contain a diluent such as physiological saline for diluting what could be a concentrated solution or lyophilized powder of the aquaporin inhibitor agent.
  • the instructions can include instructions for mixing a particular amount of the diluent with a particular amount of the concentrated pharmaceutical preparation, whereby a final formulation for injection or infusion is prepared.
  • the instructions may include instructions for treating a subject with effective amounts of the aquaporin inhibitor agent and/or instructions for treating a subject with effective amounts of an agent that increases a level of caspase activity or an agent that decreases a level of secretase activity 7 and/or decreases a level of BACE1 activity 7 .
  • the containers containing the preparations can contain indicia such as conventional markings that change color when the preparation has been autoclaved or otherwise sterilized.
  • the term “contacting” refers to any suitable method of bringing an inhibitor agent, a composition, or a pharmaceutical composition into contact with a cell.
  • any known method of administration is suitable as described herein.
  • the brains of two 6-month-old juvenile animals were examined, including two 13-month-old healthy adults and three 24 or 36- month-old spiders that showed mild (24 month), or severe (two 36-month-old animals) behavioral signs of degeneration.
  • To achieve optimal tissue preservation at both light- and ultrastructural levels the animals were deeply anesthetized with CO2. After removal of the tarsal leg segments the animals were perfused with freshly prepared ice cold 4% paraformaldehyde (EMS 15710) and 2.5% glutaraldehyde (EMS 16019) in phosphate buffered saline (pH 7.4, 0. 1 M; PBS). All procedures were in accordance with IACUC protocols.
  • peripheral structures used in this study are well-described mechanosensory neurons in the VS-3 slit sense organ that is located on the anterior part of the patella on all eight legs. Location and dissection procedure of this organ has been described previously [R. Fabian-Fine et al., The Journal of Neuroscience 19, 298-310 (1999)]. After dissection of the VS-3 organ and CNS in ice-cold fixative [R. Fabian-Fine et al., Cell and tissue research 362, 461-479 (2015)]. the preparations were washed in PBS and post-fixed for 20 min in 0.5-1.0% osmium tetroxide (Electron Microscopic Sciences, #19150).
  • the human brain samples used in the study were obtained from the brains of three decedents undergoing autopsy examination at the University of Vermont Medical Center and were fully consented for the purposes of diagnosis, research, and teaching. Samples were obtained promptly following brain removal.
  • Tissue from the decedents with a clinical history of Alzheimer dementia was obtained 14-43 hours postmortem.
  • Preparation of human brain tissue for both light and electron microscopic approaches were carried out as described for spider tissue.
  • Diagnostic neuropathologic examination of these brains revealed ADNC with an ABC score of A3B3C2, consistent with a high burden of ADNC.
  • Significant amyloid plaques and phosphorylated tau tangles were present in the hippocampus.
  • the sections were washed in PBS 4x5 min and unspecific binding sites were blocked with blocking medium containing 0.25% Bovine Serum Albumin (Sigma A4503) and 5% Normal goat serum (Sigma G9023) in 1% Triton-X/PBS for 20 min.
  • the sections were incubated overnight at 6 °C within primary rabbit anti Aquaporin4 antiserum (BiCell #20104) at a dilution of 1: 100 in PBS containing 10% blocking medium.
  • Proteins were separated using a Mini-PROTEAN TGX precast polyacrylamide gel (# 456- 1083, BIO-RAD, USA). Wells were loaded with 30 pl of supernatant and 20 pl of protein standard (BioRad # 1610399) ranging from 5-250 kDa. The gel was run at 200 V for ⁇ 30 min. Subsequently the gel was washed in distilled water 3x5 min and incubated in blotting buffer for 5 min. Loading of the blotting chamber and transfer of the proteins into nitrocellulose was performed according to the manufacturer's instructions.
  • the secondary antibody was applied for 1 h at a dilution of 1: 10,000 in the same blocking solution used for primary antibody dilution under gentle agitation, washing 4x5 min in TBS-T, immunoreactive protein bands were visualized using the BioRad Opti-4CN Substrate Kit #1708235.
  • the nitrocellulose was submerged in a solution of 1 part diluent with 9 parts of purified distilled water. After adding 0.02% of substrate the blots were agitated under visual control for ⁇ 5 min. Upon visibility of the labelled protein bands the nitrocellulose was washed in TBS-T and photographed immediately.
  • sODG A specialized characteristic of sODG is the formation of finger-like processes that project into the neuronal cytoplasm and form glial-canals of varying shapes and sizes that engulf cellular debris. Serial section analysis of images from these studies showed that these glial-canals project toward lymphatic canals that drain cellular debris from the brain.
  • Fig. 1 A-C provide a schematic diagram showing proposed glial lobe formation in spider leg ganglia.
  • sODG form two ‘lobe’ types.
  • sODG form two ‘lobe’ types.
  • the first type called ‘forming lobes’ (FLs) consists of a circular central sODG-arm around which gradually expanding circular glial membranes form. These FLs were present in all spiders observed in this study, indicative of regenerative abilities of sODG.
  • the second lobe type referred to as ‘mature lobes’ consists of numerous glial rings that are compacted laterally and take on an elongated appearance whereby the membranes of individual glial rings are compacted laterally and appear linearly aligned. During this compaction, glial cytoplasm and organelles are pushed toward the lateral boundaries. Besides mitochondria, glial lobes contain two conspicuous structures.
  • glial aqua canals were translocated into the neuronal cytoplasm (neuronal aqua canals) and created a cytoplasmic bulk flow that caused swelling of the neuronal aqua canals and the flow of cellular debris into the lumina of the glial-canals.
  • GACs glial aqua-canals'
  • NACs neuroneuronal aqua-canals
  • Forming glial-canals were associated with both GACs and NACs and accumulated large waste clusters (up to > 2 pm in diameter) in or around their lumina.
  • the internalization of waste into the glial cells was either through phagocytosis or through engulfment by one or more glial membranes.
  • the distal tips of canal-forming glial membranes often appeared bulbous and contained GAC profiles that formed openings into the neuronal cytoplasm. Networks of membrane cistemae were located around these distal tips.
  • the formation of debris-containing glial-canals was observed in animals of all developmental stages suggesting that their formation is a normal process by which macroglia ensure controlled waste removal from neurons.
  • AQP4-immunolabeling was used. Strong AQP4-LIR was observed within myelinated glial- canals of varying diameters that are located alongside or within neuronal somata and were distinguishable by their unstructured lumina. These structures were referred to as (tanycyte- like) ‘glial-canals.’ AQP4-LIR profiles within the neuronal somata were consistent with the location, shape, and size of both NACs and GACs.
  • AQP4-LIR was detected in (a) cells lining a tubular system that borders dorsally onto the CNS and is continuous with the gastrointestinal tract (b) tubular appearing glial cells within the brain parenchyma and leg nerves, and (c) in mesenchymal cells in the neural lamella.
  • Advanced degeneration resulted in the rupture of sODG lobes that separated the neuronal cytoplasm from adjacent tanycyte-like glial-canals facilitating the complete cytoplasmic evacuation from neurons.
  • the term ‘gliaptosis' was used to describe this glial-induced neuronal cell death via evacuation of cytoplasmic content by adjacent macroglia.
  • the dorsal tubular system was identified as taking on a brown discoloration consistent with this observed in nephrocytes that internalize cellular debris in degenerating brains.
  • Neuronal cell death w as follow ed by the disintegration of surrounding sODG and gradual replacement with particulate matter as described previously [R. Fabian- Fine et al.. Journal of Comparative Neurology 531, 618-638 (2023)].
  • Vimentin/AQP4 double labelling enabled differentiation of tanycyte profiles from AQP4- negative oligodendroglial and neuronal processes. These cell processes are referred to as tanycytes.
  • Semithin sections showed the formation of swelling varicose projections by tanycytes that were often in close association with cellular waste.
  • human AQP4-LIR tanycytes that project from the ventricular lining into hippocampal neurons form numerous circular fenestrated cell structures, which attach to adjacent cells, likely forming stabilizing ‘connective junctions’ between the cells.
  • tanycytes At the ultrastructural level, tanycytes have an inner lumen that is separated from the peri-luminal space.
  • Varicose projections could be seen to emanate from this peri -luminal space projecting into adjacent cells. Such intraneuronal projections were often adjacent to or surrounded by a ‘penumbra’ of clear cytoplasm. To assess whether the function of these varicosities is to create an AQP4- mediated cytoplasmic bulk flow toward tanycytes and ensure waste removal from neurons, studies were conducted that included vimentin AQP4 double labeling. Tanycytes stained vimentin positive in the central lumen, whereas the peri-luminal space together with emanating varicose projections appeared strongly AQP4-LIR supporting the hypothesis.
  • these glial-canals may be mistaken for mitochondrial profiles if the connecting glial stalk is outside of the section plane.
  • these glial projections formed prominent membrane cistemae that projected into the neuronal cytoplasm similar to this observed in central spider neurons.
  • Results of studies support that a role of these membrane cistemae in endocytotic waste uptake from the neuronal cytoplasm into surrounding glial cells.
  • degenerating human tanycyte profiles form numerous swelling projections that transect from the peri-luminal space into adjacent cells.
  • neuronal profiles adjacent to degenerating tanycyte profiles appeared to gradually clear of their cytoplasmic content.
  • Results of experiments described herein demonstrate that the structural failure of this canal system caused the catastrophic depletion of cy toplasmic content from affected neurons leading to a previously undescribed type of macroglia-induced cell death, which is referred to herein as 'gliaptosis/
  • Studies set forth herein demonstrated that the cellular characteristics of neuronal cell death in the brain of Alzheimer patients with ADNC resemble those described in spider neurons and provide compelling evidence that AQP4-LIR tanycyte profdes may form a similar AQP4 water channel -mediated waste clearance system as this proposed for spider neurons.
  • Results show that tanycytes in degenerating human brain undergo hypertrophic changes and provide strong evidence for an AQP4 water channel -mediated catastrophic depletion of cytoplasmic content from affected neurons similar to the mechanism observed in spider brain. Based on evidence obtained from experiments described herein it is proposed that these pathological changes in tanycytes and the resulting impairment of the glial-canal system may be one of the underlying causes for neuronal cell death in the brain of Alzheimer patients with ADNC.
  • the two key components that form the structural and functional foundation for these canals are (a) myelin, and (b) AQP4-LIR glial cells that transect from the ventricular lining into the hippocampal formation.
  • myelin is the formation of intraneuronal projections and membrane cistemae that are important for the uptake and removal of cellular waste via endocytosis, phagocytosis, or engulfment of debris by glial membranes.
  • myelination in invertebrate neurons is considered unusual [D. K. Hartline and D. R. Colman, Current Biology 17, R29-R35 (2007); A. I. Boulleme, Experimental Neurology 283, 431-445 (2016); B. I.
  • the glial-canal system described herein although structurally altered- appears highly conserved between arachnids and mammals and alludes to the vital importance of this system for the functional integrity and survival of neurons.
  • the mammalian brain Compared to arachnids, the mammalian brain has evolved favoring increased numbers of (a) neurons, (b) dendritic arbors, and (c) synaptic connectivity in exchange for structurally smaller neurons and glial cells. It is likely due to this evolutionary trend and spatial limitations that waste clearance mechanisms in mammals have been reduced to myelinated tanycyte projections along neuronal somata and neurites compared to the space-occupying peri-somatic myelination in spiders.
  • mammalian tanycytes internalize catabolized cell waste that is likely broken down by lytic enzymes contained in and around tanycyte waste receptacles .
  • mammalian tanycyte projections into neurons are often adjacent to or surrounded by a penumbra in which the cytoplasm appears clear and less structured.
  • structurally impaired tanycytes whose outer myelin sheath unravels and opens to the extracellular space show a similar penumbra in which cells that border on impaired tanycytes appear to gradually lyse.
  • microtubule associated proteins H. V. Goodson and E. M. Jonasson, Cold Spring Harbor perspectives in biology 10, a022608 (2016)].
  • Such interactions include motor proteins [H. L. Sweeney and E. L. Holzbaur, Cold Spring Harbor Perspectives in Biology 10, a021931 (2016)], membrane-bound organelles [G. Kanfer et al., Molecular biolog/ of the cell 28, 2400-2409 (2017); A. J. Lomakin et d ., Molecular biology of the cell 22, 4029-4037 (2011)], chromosomes [N. B. Gudimchuk and J. R. McIntosh, Nature reviews Molecular cell biology 22.
  • the human brain tissue used here was obtained from four decedents undergoing autopsy examination at the University of Vermont Medical Center with full consent for biomedical research, diagnosis, and teaching purposes according to Vermont State law. The samples were fixed immediately after removal of the brains.
  • the sections were kept in blocking medium consisting of 0.25% Bovine Serum Albumin (Sigma A4503) and 5% Normal Goat Serum (Sigma G9023) in 1% Triton-X/PBS for 20 min. Incubation with the primary goat anti-rabbit aquaporin-4 antiserum (BiCell #20104) and goat-anti mouse anti-Vimentin (#AMF-17b-s) at a dilution of 1 : 100 in PBS containing 10% blocking medium overnight. Both antibodies are routinely used in human tissue. Subsequently the sections were washed for seven wash cycles in PBS prior to an additional 20-min blocking step in blocking medium.
  • blocking medium consisting of 0.25% Bovine Serum Albumin (Sigma A4503) and 5% Normal Goat Serum (Sigma G9023) in 1% Triton-X/PBS for 20 min. Incubation with the primary goat anti-rabbit aquaporin-4 antiserum (BiCell #20104) and goat-anti mouse anti-V
  • the secondary' fluorochrome-coupled antibodies (Cy3 goat anti-rabbit, Jackson ImmunoResearch Laboratories 111-165-003. FITC goat anti-mouse Jackson ImmunoResearch Laboratories 115-096-072) were used at a dilution of 1:600 in PBS containing 10% blocking medium overnight in the fridge.
  • the sections were washed in PBS for three washing cycles prior to staining with Hoechst Blue nuclear stain (Sigma H 6024; 1 :3000 in PBS) for 20 min. After washing in PBS for five wash cycles the sections were mounted on glass slides and embedded in Mowiol (Sigma# 81381). Due to the light sensitivity of the secondary antibodies all steps were conducted under minimum light conditions to prevent bleaching of the fluorochromes.
  • the sections were analyzed using a confocal Zeiss Axiolmager MZ with Apotome.
  • Preparations processed for ultrastructural analysis were fixed in a mixture of 4% paraformaldehyde and 2.5% glutaraldehyde (EMS 16019) in PBS overnight in the fridge. It is important to note that incubation in this fixative for longer time periods as described here are detrimental to histological staining procedures as semithin sections of the tissue fixed for extended time periods does not stain sufficiently for histological dyes.
  • the preparations were post-fixed in 1.0% osmium tetroxide (Electron Microscopic Sciences, #19150) for approximately 1 hour. After two wash cycles in PBS, the tissue was dehydrated in a graded series of molecular grade ethanol.
  • the preparations were promptly transferred into propylene oxide (Electron Microscopic Sciences, #20401) and slowly infiltrated with Araldite (Electron Microscopic Sciences, #13900). It is important to note that the extended exposure (1-3 days) of the osmicated tissue to ethanol and propylene oxide may result in the removal of osmium from the tissue and compromise tissue preservation. It is thus important not to leave the samples in these media overnight.
  • the tissue was embedded in pure Araldite and polymerized overnight at 60 °C, according to manufacturer's instructions. Semi- and Ultrathin Sectioning.
  • Vibratome sections from 4% paraformaldehyde-fixed tissue were washed in PBS for four wash cycles. Staining with toluidine blue was conducted under visual control by slowly dripping a 2% toluidine blue solution into a petri dish containing brain sections in PBS. The staining was terminated when the neurons appeared appropriately stained for light microscopic investigation without overstaining the preparations. Overstaining will result in the inability to distinguish individual structures in the section.
  • the sections were embedded in Mowiol and examined promptly using an Olympus microscope with differential phase contrast and digital image capturing capabilities. The embedding medium will slowly de-stain the cells, however cellular debris within the cells remains clearly visible in form of brown deposits.
  • Brains were emersed in 10% neutral buffered formalin and incubated for at least 1 week at room temperature. Hippocampal tissue was removed from the brains, dehydrated in a graded series of ethanol, infiltrated with 100% xylene (3x 40 min), and embedded in paraffin (25 min). Sections were cut at 10 mm thickness and mounted on glass slides. Slides were rinsed in xylene (2x), 100% and 95% EtOH (lx each) prior to immersion in 0.1% Luxol fast blue solution (Leica #) at 60 °C overnight at 60 degrees C. Excess stain was removed with 95% EtOH. Slides were rinsed in distilled water and placed in a 0.05% lithium carbonate solution (2 min) followed by 70% EtOH (2 min).
  • Tanycytes form varicose protrusions into hippocampal neurons and glial cells
  • AD-decedents Light microscopic analysis of toluidine-blue stained vibratome sections through the hippocampal formation of patients with diagnosed Alzheimer disease neuropathologic change (referred to in the following as ‘ AD-decedents’) reveals that neuronal somata and their initial axon segments are contacted by numerous varicose cell processes that form large numbers of bulbous intraneuronal protrusions that are associated with cellular debris. These structures are commonly assumed to represent lipofuscin. Double labeling for AQP4 and vimentin indicate an ependymal origin of these cell processes .
  • Pyramidal cells from decedents negative for Alzheimer disease neuropathologic change showed fewer and smaller protrusions within the cytoplasm and less debris accumulation around intraneuronal protrusions.
  • Investigation of both human hippocampus and olivary nucleus using Luxol blue stain show vast numbers of myelinated tanycyte processes transecting from the ventricular lining into the brain parenchyma.
  • Immunolabeling for AQP4 shows strong AQP4-immunoreactivity (AQP4-IR) within these ependymal processes.
  • Circular myelin-derived protrusions that transect into the neuronal cytoplasm mature into specialized receptacles that internalize electron dense cellular debris into a centrally located myelinated canal.
  • AQP4-IR tanycytes form ‘adhesion-clamps ’ and protrusions into adjacent cell profiles
  • Tanycyte processes form AQP4-IR circular clamp-like structures whose diameters range between ⁇ 1 and 10 pm. Individual clamps surround cellular processes of adjacent cells. These circular structures are referred to herein as "adhesion clamps’ (ACs). In AD-affected tissue ACs form numerous protrusions and could be seen swelling to large diameters of >10 pm. Immunolabeling for AQP4 and vimentin demonstrated that these clamps formed along the processes of ependymal tanycytes whose somata reside in the ventricular lining.
  • AQP4-IR tanycytes were observed projecting into the stratum pyramidale forming numerous protrusions alongside and within neuronal somata and astrocytes consistent with the protrusions observed in toluidine-blue stained vibratome sections.
  • tanycytes were observed forming numerous projections along astrocytes whose somata appeared unstained for AQP4 compared to the somata of tanycytes.
  • H&E-stained tanycyte processes in the alveus further show their varicose and inter-connected nature consistent with the ultrastructural observations.
  • Vimentin/ AQP4-double labeling demonstrated that AQP4-IR protrusions that transect toward cell nuclei emanate from vimentin-IR fiber profiles consistent with the observations that AQP4-expressing tanycytes form cellular protrusions into somata.
  • Type-A protrusions were predominantly located within the neuronal cytoplasm and consisted of the central canal that appeared electron dense that was surrounded by several electron lucent bulbous structures (in the following referred to as ‘receptacles’) that have small openings to the neuronal cytoplasm (in the following referred to as ‘outer gates’).
  • the receptacles were separated from the central canal via perforated membranes, referred to herein as ’inner diaphragm.’ Interestingly, numerous vesicular structures approx.
  • Type-B The second protrusion type was predominantly observed in extracellular spaces. This type consisted of a central canal that contained numerous membrane folds within its lumen. In this type, debris-filled central canals were observed around the receptacle. Characteristic circular openings resembling those in Type A receptacles were observed. Interestingly, Type-B protrusions contained numerous distinct circular structures that were near electron-dense cellular debris.
  • Fig. 3A-S shows results of studies in which sections through the hippocampus of decedents with diagnosed AD were assessed. De-stained hippocampal sections of AD patients (in which cellular debris appeared brown stained) showed abundant brown coloration within most brain areas (Fig. 3). Particularly dense accumulations were observed (1) within neuronal somata in the stratum pyramidale, (2) the stratum oriens, and (3) the alveus.
  • Fig. 3A provides photomicrographic image showing areas investigated, which included the alveus, stratum oriens and CAI -C A3 stratum pyramidale (Luxol H&E stain). Fig.
  • Fig. 3H-K provides images of different focal planes of a stained tanycyte process that gave rise to circular protrusions.
  • the image in Fig. 3L shows hypertrophic Luxol blue-stained tanycyte process that contained engorged translucent compartments consistent with excessive liquid intake. Adjacent tanycyte processes appeared less engorged.
  • 3M is an image showing neuronal soma in the olivary nucleus of an AD-decedent densely obstructed with hypertrophic tanycyte receptacles. Results demonstrated that hypertrophic tanycyte processes contacted the neuronal cytoplasm and nucleus and showed tanycyte processes transecting into the neuronal cytoplasm.
  • the Fig. 3N image shows an H&E-stained hypertrophic tanycyte projection in the alveus in close proximity to brown cellular waste and Fig. 30 shows tanycyte soma with signs of hypertrophy onset in the soma. Images at different focal planes through the soma of a hypertrophic tanycyte soma with attached tanycyte processes are shown in Fig. 3P-S.
  • Fig. 4B provides a schematic diagram of a mechanism of waste uptake into tanycyte receptacles.
  • Receptacles are formed through the formation of myelin protrusions around the outer periphery' of myelinated tanycytes.
  • the central, AQP4-expressing canal branches into each receptacle-forming protrusion.
  • Central canal and protrusion form a functional unit by which the functional significance of the receptacle is the filtering and catabolism of cellular waste to prevent obstruction by larger debris particles.
  • results of the studies set forth herein demonstrated that tanycytes interacted with neurons, glial cells, and extracellular spaces via specialized receptacles, likely to ensure controlled waste removal from the brain.
  • the results support a conclusion that hypertrophic abnormalities result in functionally compromised tanycytes.
  • the resulting obstruction of neurons by hypertrophic tanycyte receptacles may cause neuronal cell death in affected neurons.
  • one cause for hypertrophic abnormalities in affected tanycytes appears to be the physical obstruction of apical drainage canals due to abnormal waste aggregation.
  • Results indicate: (a) debris is flushed toward these receptacles via the formation of an AQP4-mediated bulk flow, (b) internalized into central canals, (c) flushed toward the ventricular lining as well as paravascular spaces and (d) drained into CSF. Results suggest that obstruction of this canal system in apical drainage canals may cause an imbalance whereby waste uptake exceeds waste drainage. The resulting increase in turgor within affected tanycytes explains the hypertrophic swelling of myelin protrusions, and spongiform pathologies in the brains of AD- decedents described here. Results indicate that the large number of affected brain cells may be explained by the reticular, interconnected nature of these ependymal cells and their processes.
  • Tanycytes are ependymal glial cells located in the ventricular lining that send long slender processes into the brain parenchyma. Prior knowledge regarding these glial cells is only fragmentary' and the significance of the myelinated processes they form within the brain parenchyma has not been recognized.
  • Currently four different tanycyte types are distinguished based on their location in the third ventricle [R. Dali et al., Physiol. Behav. 263, 1 14108 (2023); R. Pasquettaz et al., J. Comp. Neurol. 529, 553 (2021 )]. However, it has been proposed that more types may exist [Z.
  • Results presented herein demonstrate close cell interactions between tanycytes with neurons and astrocytes. Results support a conclusion that the primary purpose of these interactions may be waste removal from metabolically active cells. Results of studies disclosed herein indicate that a functional foundation for tanycyte-mediated waste clearance in the human brain depends on: (a) the process by which myelinated tanycytes transect into adjacent cells, (b) the ability of myelinated tanycytes to form serial bulbous protrusions, that (c) differentiate into specialized receptacles for waste uptake, (d) the proposed AQP4- mediated flow toward tanycyte receptacles, and (e) proteolytic enzy mes that catabolize larger debris particles, thus preventing blockage of the long, narrow tanycyte processes.
  • Results presented herein relating to spider brain support the proposed role of myelinated glial cells that transect into the neuronal cytoplasm to engulf and remove cellular waste from the healthy brain.
  • C. salei also shows progressive neurodegeneration and degeneration onset has similarly been linked to hypertrophic abnormalities of myelinated glial cells [R. Fabian-Fine et al., J. Comp. Neurol. 531, 618-638 (2023)].
  • results presented herein provide evidence supporting a conclusion that structural impairment and resulting hypertrophy of tanycytes may be one of the main causes for neurodegeneration.
  • This type of glia-mediated cell death characterized by the depletion of cytoplasmic content into adjacent glial cells is referred to herein as “gliaptosis.”
  • structurally impaired glia cells in spiders showed similar hypertrophic swelling, discoloration and depletion of neuronal content shown here in AD affected brain [R. Fabian-Fine et al.. J. Comp. Neurol. 531, 618-638 (2023)].
  • AQP4 The prevalence of AQP4 in the brain has mainly been attributed to their expression in the end feet of astrocytes [S. Wang et al., Aquaporins, Springer, pp. 317-330 (2023); K. Oshio et al.. Neuroscience 127, 685-693 (2004); Q. Lu et al., Int. J. Biol. Sci. 18, 441-458 (2022)].
  • astrocytes are abundant in the stratum oriens, adjacent to the stratum radiatum from which large numbers of AQP4-IR tanycyte processes emerge. If astrocytes indeed synthesize large numbers of AQP4 one would expect strongly immunoreactive somata, reflective of the place of protein synthesis. However, as shown in results herein, the somata of astrocytes did not show significant immunolabeling for AQP4.
  • Results presented herein support a conclusion that the obstruction of apical drainage canals should be considered, and that tanycytes whose apical waste-drainage canals are blocked lose the ability to effectively drain debris that is taken up at their basal ends.
  • the intracellular turgor increases which explains the hypertrophic appearance of (a) tanycyte somata in the ventricular lining, (b) ACs that are formed by affected tanycytes, (c) spongiform abnormalities within and around tanycyte processes, and (d) the excessive formation and swelling of both intra- and extracellular tanycyte receptacles in neurons that are connected to affected tanycytes.
  • Results presented herein support a conclusion that the controlled downregulation of AQP4 channels and/or increase activity of secretases and/or BACE1, would allow affected tanycytes to restore balance between water uptake and drainage and reduce hypertrophy of both affected tanycytes and neurons whose somata are obstructed.
  • the structural appearance of the receptacles in human brain described herein is likely specialized to prevent larger debris particles from blocking the long and narrow mammalian tanycyte processes.
  • the narrow openings of the receptacles are lined by membranous structures that resemble enzyme complexes.
  • the combination of narrow openings surrounded by proteolytic enzymes may prevent larger particles from entering the receptacles.
  • Type- A receptacles described herein are formed within the cytoplasm of cells and may be triggered by intracellular signals inherent to catabolic processes. Results of studies presented herein, indicated that Type-B receptacles were predominantly found in extracellular spaces, supporting a conclusion that they play an important role in extracellular waste clearance.

Landscapes

  • Health & Medical Sciences (AREA)
  • Neurosurgery (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Hospice & Palliative Care (AREA)
  • Psychiatry (AREA)
  • Psychology (AREA)
  • Epidemiology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne des compositions et des procédés pour prévenir la mort cellulaire neuronale. Les procédés de l'invention concernent, en partie, la réduction d'une fonction anormale d'un canal glial dans une cellule gliale de formation de canal adjacente à une cellule neuronale.
PCT/US2024/061013 2023-12-20 2024-12-19 Procédés et compositions pour réduire la gliaptose : mort cellulaire neuronale induite par macroglie Pending WO2025137265A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US202363612602P 2023-12-20 2023-12-20
US63/612,602 2023-12-20
US202463644708P 2024-05-09 2024-05-09
US63/644,708 2024-05-09

Publications (1)

Publication Number Publication Date
WO2025137265A1 true WO2025137265A1 (fr) 2025-06-26

Family

ID=94322990

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2024/061013 Pending WO2025137265A1 (fr) 2023-12-20 2024-12-19 Procédés et compositions pour réduire la gliaptose : mort cellulaire neuronale induite par macroglie

Country Status (1)

Country Link
WO (1) WO2025137265A1 (fr)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004053144A2 (fr) * 2002-12-10 2004-06-24 Ottawa Health Research Institute Modulation de la differenciation de cellules souches par modulation de l'activite de la caspase-3
US20070281978A1 (en) * 2006-06-01 2007-12-06 Niigata University Inhibitors of aquaporin 4, methods and uses thereof
KR20180020718A (ko) * 2016-08-19 2018-02-28 동아대학교 산학협력단 이소플라본을 유효성분으로 포함하는 퇴행성 뇌질환의 예방 또는 치료용 조성물
US20190127360A1 (en) 2016-04-28 2019-05-02 Apoglyx Ab Compounds for modulating aquaporins
WO2021260387A1 (fr) * 2020-06-26 2021-12-30 The University Of Birmingham Inhibition de mmp-9 et mmp-12 pour le traitement d'une lésion de la moelle épinière ou d'une lésion associée à un tissu neurologique

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004053144A2 (fr) * 2002-12-10 2004-06-24 Ottawa Health Research Institute Modulation de la differenciation de cellules souches par modulation de l'activite de la caspase-3
US20070281978A1 (en) * 2006-06-01 2007-12-06 Niigata University Inhibitors of aquaporin 4, methods and uses thereof
US20190127360A1 (en) 2016-04-28 2019-05-02 Apoglyx Ab Compounds for modulating aquaporins
KR20180020718A (ko) * 2016-08-19 2018-02-28 동아대학교 산학협력단 이소플라본을 유효성분으로 포함하는 퇴행성 뇌질환의 예방 또는 치료용 조성물
WO2021260387A1 (fr) * 2020-06-26 2021-12-30 The University Of Birmingham Inhibition de mmp-9 et mmp-12 pour le traitement d'une lésion de la moelle épinière ou d'une lésion associée à un tissu neurologique

Non-Patent Citations (87)

* Cited by examiner, † Cited by third party
Title
A. 1. BOULLERNE, EXPERIMENTAL NEUROLOGY, vol. 283, 2016, pages 431 - 445
A. D. DAVIS ET AL., NATURE, vol. 398, 1999, pages 571
A. J. LOMAKIN ET AL., MOLECULAR BIOLOGY OF THE CELL, vol. 22, 2011, pages 4029 - 4037
A. J. SMITH ET AL., ELIFE, 2017, pages 6
A. MORENO-GARCIA ET AL., FRONT NEUROSCI, vol. 12, 2018, pages 464
A. PETERS ET AL.: "The fine structure of the nervous system : neurons and their supporting cells", 1991, OXFORD UNIVERSITY PRESS
A. RUIZ ET AL., NEURON, vol. 39, 2003, pages 961 - 973
A. VAN HARREVELD, THE STRUCTURE AND FUNCTION OF NERVOUS TISSUE, vol. 4, 1972, pages 447 - 511
C. STADELMANN ET AL., PHYSIOL. REV., vol. 99, 2019, pages 1381 - 1431
CATALIN BOGDAN ET AL: "Cerebrolysin and Aquaporin 4 Inhibition Improve Pathological and Motor Recovery after Ischemic Stroke", CNS & NEUROLOGICAL DISORDERS, vol. 17, no. 4, 6 July 2018 (2018-07-06), NL, pages 299 - 308, XP093263637, ISSN: 1871-5273, Retrieved from the Internet <URL:https://eurekaselect.com/article/download/161549> DOI: 10.2174/1871527317666180425124340 *
CUI DAN ET AL: "Alleviation of Cerebral Infarction of Rats With Middle Cerebral Artery Occlusion by Inhibition of Aquaporin 4 in the Supraoptic Nucleus", ASN NEURO, vol. 12, no. 1, 1 January 2020 (2020-01-01), pages 1 - 14, XP093263608, ISSN: 1759-0914, Retrieved from the Internet <URL:http://journals.sagepub.com/doi/full-xml/10.1177/1759091420960550> DOI: 10.1177/1759091420960550 *
D. K. HARTLINED. R. COLMAN, CURRENT BIOLOGY, vol. 17, 2007, pages R29 - R35
D. LEGLAND ET AL., BIOINFORMATICS, vol. 32, 2016, pages 3532 - 3534
E. E. CONGDONE. M. SIGURDSSON, NATURE REVIEWS NEUROLOGY, vol. 14, 2018, pages 399 - 415
E. FARKAS ET AL., ISCIENCE, 2020, pages 23
E. HORSTMANNH. MEVES, ZEITSCHRIFTFÜR ZELLFORSCHUNG UNDMIKROSKOPISCHE ANATOMIE, vol. 49, 1959, pages 569 - 604
E. RODRIGUEZ ET AL., JOURNAL OF NEUROENDOCRINOLOGY, 2019, pages 31
F. DOETSCH ET AL., JOURNAL OF NEUROSCIENCE, vol. 17, 1997, pages 5046 - 5061
G. KANFER ET AL., MOLECULAR BIOLOGY OF THE CELL, vol. 28, 2017, pages 2400 - 2409
G. M. PRESTON ET AL., SCIENCE, vol. 256, 1992, pages 385 - 387
G. SEIFERTC. STEINHÄUSER, CELL AND TISSUE RESEARCH, vol. 373, 2018, pages 653 - 670
H. F. CSERRL. H. OSTRACH, EXPERIMENTAL NEUROLOGY, vol. 45, 1974, pages 50 - 60
H. L. SWEENEYE. L. HOLZBAUR, COLD SPRING HARBOR PERSPECTIVES IN BIOLOGY, vol. 10, 2018, pages a021931
H. S. SAWHNEYC. P. PATHAKJ. A. HUBELL, MACROMOLECULES, vol. 26, 1993, pages 581 - 587
H. V. GOODSONE. M. JONASSON, COLD SPRING HARBOR PERSPECTIVES IN BIOLOGY, vol. 10, 2018, pages a022608
IGARASHI HIRONAKA ET AL: "Pretreatment with a novel aquaporin 4 inhibitor, TGN-020, significantly reduces ischemic cerebral edema", NEUROLOGICAL SCIENCES (TESTO STAMPATO), vol. 32, no. 1, 6 October 2010 (2010-10-06), IT, pages 113 - 116, XP093263665, ISSN: 1590-1874, DOI: 10.1007/s10072-010-0431-1 *
J. EBLINGJ. E. LEWIS, GLIA, vol. 66, 2018, pages 1176 - 1184
J. J. HARTMANR. D. VALE, SCIENCE, vol. 286, 1999, pages 782 - 785
J. J. ILIFF ET AL., SCIENCE TRANSLATIONAL MEDICINE, vol. 4, 2012
J. J. ILIFFM. NEDERGAARD, STROKE, vol. 44, 2013, pages S93 - S95
J. J. ILIFFM. NEDERGAARD, STROKE, vol. 44, pages S93 - S95
J. SCHINDELIN ET AL., NATURE METHODS, vol. 9, 2012, pages 676 - 682
K. 1. VOUMVOURAKIS ET AL., BIOMEDICINES, vol. 11, 2023, pages 2092
K. A. JELLINGERA. D. KORCZYN, BMC MED, vol. 16, 2018, pages 34
K. OSHIO ET AL., NEUROSCIENCE, vol. 127, 2004, pages 685 - 693
K. WEBER ET AL., EUR. J. CELL BIOL., vol. 101, 2022, pages 151218
KATANO T ET AL., DRUG METAB PHARMACOKINET, vol. 29, no. 4, 2014, pages 348 - 51
L. A. LIGON ET AL., NATURE CELL BIOLOGY, vol. 3, 2001, pages 913 - 917
L. C. WALKER ET AL., JAMA NEUROLOGY, vol. 70, 2013, pages 304 - 310
L. KIANI, NATURE REVIEWS NEUROLOGY, vol. 19, 2023, pages 459 - 459
L. SOLNICA-KREZELW. DRIEVER, DEVELOPMENT, vol. 120, 1994, pages 2443 - 2455
LU QI ET AL: "Minocycline improves the functional recovery after traumatic brain injury via inhibition of aquaporin-4", INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES, vol. 18, no. 1, 1 January 2022 (2022-01-01), pages 441 - 458, XP093263598, ISSN: 1449-2288, DOI: 10.7150/ijbs.64187 *
M. AMIRY-MOGHADDAMO. P. OTTERSEN, NATURE REVIEWS NEUROSCIENCE, vol. 4, 2003, pages 991 - 1001
M. ASKENAZI ET AL., NATURE COMMUNICATIONS, vol. 14, 2023, pages 2902 - 15
M. BOLBOREAN. DALE, TRENDS IN NEUROSCIENCES, vol. 36, 2013, pages 91 - 100
M. BRIGHTMAN ET AL., THE JOURNAL OF CELL BIOLOGY, vol. 40, 1969, pages 648 - 677
M. KIRSCHNERT. MITCHISON, CELL, vol. 45, 1986, pages 329 - 342
M. NEDERGAARD, SCIENCE, vol. 340, 2013, pages 1529 - 1530
M. NEDERGAARDS. A. GOLDMAN, SCIENTIFIC AMERICAN, vol. 314, 2016, pages 44 - 49
M. OKLINSKI ET AL., J. HISTOCHEM. CYTOCHEM., vol. 62, 2014, pages 598 - 611
M. SCHWEIGHAUSER ET AL., NATURE, vol. 585, 2020, pages 464 - 469
M. SIMONSK.-A. NAVE, COLD SPRING HARB. PERSPECT. BIOL, vol. 8, 2015
N. B. GUDIMCHUKJ. R. MCINTOSH, NATURE REVIEWS MOLECULAR CELL BIOLOGY, vol. 22, 2021, pages 777 - 795
N. J. ABBOTT ET AL., ACTA NEUROPATHOL, vol. 135, 2018, pages 387 - 407
N. J. ABBOTT ET AL., ACTA NEUROPATHOLOGICA, vol. 135, 2018, pages 387 - 407
N. J. ABBOTT ET AL., ACTCL NEUROPATHOLOGICA, vol. 135, 2018, pages 387 - 407
N. L. HAUGLUND ET AL., CURRENT OPINION IN PHYSIOLOGY, vol. 15, 2020, pages 647 - 665
N. SAKR ET AL., INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 24, 2023, pages 3432
P. A. ADENIYI ET AL., ASN NEURO, vol. 14, no. Sage Publications, Ltd., 2022, pages 1 - 13
P. AGRE ET AL., JOURNAL OF PHYSIOLOGY, vol. 542, 2002, pages 3 - 16
P. N. DE FRANCESCO ET AL., CELL AND TISSUE RESEARCH, vol. 369, 2017, pages 369 - 380
P. W. BAAS ET AL., CYTOSKELETON, vol. 73, 2016, pages 442 - 460
PERGAKIS MELISSA ET AL: "An update on the pharmacological management and prevention of cerebral edema: current therapeutic strategies", EXPERT OPIN PHARMACOTHER, vol. 22, no. 8, 27 January 2021 (2021-01-27), London, UK, pages 1025 - 1037, XP093263278, ISSN: 1465-6566, DOI: 10.1080/14656566.2021.1876663 *
Q. LU ET AL., INT. J. BIOL. SCI., vol. 18, 2022, pages 441 - 458
R. DALI ET AL., PHYSIOL. BEHAV, vol. 263, 2023, pages 114108
R. DALI ET AL., PHYSIOLOGY & BEHAVIOR, vol. 263, 2023, pages 114108
R. FABIANE.-A. SEYFARTH, CELL AND TISSUE RESEARCH, vol. 287, 1997, pages 413 - 423
R. FABIAN-FINE ET AL., CELL AND TISSUE RESEARCH, vol. 362, 2015, pages 461 - 479
R. FABIAN-FINE ET AL., J. COMP. NEUROL, vol. 531, 2023, pages 618 - 638
R. FABIAN-FINE ET AL., J. COMP. NEUROL., vol. 531, 2023, pages 618 - 638
R. FABIAN-FINE ET AL., JOURNAL OF COMPARATIVE NEUROLOGY, vol. 531, 2023, pages 618 - 638
R. FABIAN-FINE ET AL., THE JOURNAL OF COMPARATIVE NEUROLOGY, vol. 420, 2000, pages 195 - 210
R. FABIAN-FINE ET AL., THE JOURNAL OF NEUROSCIENCE, vol. 19, 1999, pages 298 - 310
R. PASQUETTAZ ET AL., J. COMP. NEUROL, vol. 529, 2021, pages 553
R. PASQUETTAZ ET AL., THE JOURNAL OF COMPARATIVE NEUROLOGY, vol. 529, 2021, pages 553
S. B. HLADKYM. A. BARRAND, FLUIDS AND BARRIERS OF THE CNS, vol. 19, 2022, pages 1 - 33
S. ECKERLE ET AL., DEVELOPMENTAL BIOLOGY, vol. 434, 2018, pages 249 - 266
S. POCKES ET AL., TRANSLATIONAL RESEARCH, vol. 254, 2023, pages 34 - 40
S. WANG ET AL.: "Aquaporins", 2023, SPRINGER, pages: 317 - 330
SUN CHENGFENG ET AL: "Acutely Inhibiting AQP4 With TGN-020 Improves Functional Outcome by Attenuating Edema and Peri-Infarct Astrogliosis After Cerebral Ischemia", FRONTIERS IN IMMUNOLOGY, vol. 13, 3 May 2022 (2022-05-03), Lausanne, CH, XP093263508, ISSN: 1664-3224, DOI: 10.3389/fimmu.2022.870029 *
T. M. O'HERRON ET AL., NEUROBIOLOGY OF AGING, vol. 21, 2000, pages 145 - 145
V. PREVOT ET AL., ENDOCRINE REVIEWS, vol. 39, 2018, pages 333 - 368
VERMA N ET AL., NATURE COMMUNICATIONS, vol. 6, no. 1, 2015, pages 6064
W. MOBIUS ET AL., BRAIN RES, vol. 1641, 2016, pages 92 - 100
W. MÖBIUS ET AL.: "Methods Cell Biol.", vol. 96, 2010, ELSEVIER, pages: 475 - 512
Y.-W. KUOJ. HOWARD, TRENDS IN CELL BIOLOGY, vol. 31, 2021, pages 50 - 61
YUKUTAKE Y ET AL., BIOCHEMISTRY, vol. 48, no. 51, 29 December 2009 (2009-12-29), pages 12059 - 61

Similar Documents

Publication Publication Date Title
Hendricson et al. Aberrant synaptic activation of N-methyl-D-aspartate receptors underlies ethanol withdrawal hyperexcitability
Amartumur et al. Neuropathogenesis-on-chips for neurodegenerative diseases
Ren et al. Omega‐3 polyunsaturated fatty acids promote amyloid‐β clearance from the brain through mediating the function of the glymphatic system
Kook et al. Aβ1–42-RAGE interaction disrupts tight junctions of the blood–brain barrier via Ca2+-calcineurin signaling
Frautschy et al. Phenolic anti-inflammatory antioxidant reversal of Aβ-induced cognitive deficits and neuropathology
Sivak The aging eye: common degenerative mechanisms between the Alzheimer's brain and retinal disease
Kaindl et al. Antiepileptic drugs and the developing brain
Spoleti et al. Dopamine neuron degeneration in the Ventral Tegmental Area causes hippocampal hyperexcitability in experimental Alzheimer’s Disease
Samantaray et al. Chronic intermittent ethanol induced axon and myelin degeneration is attenuated by calpain inhibition
Caldero et al. Development of microglia in the chick embryo spinal cord: implications in the regulation of motoneuronal survival and death
Valdez et al. Retinal microangiopathy in a mouse model of inducible mural cell loss
Fu et al. Ectopic vesicular glutamate release at the optic nerve head and axon loss in mouse experimental glaucoma
Wijesinghe et al. Ergothioneine, a dietary antioxidant improves amyloid beta clearance in the neuroretina of a mouse model of Alzheimer’s disease
Wu et al. Amyloid fibril–induced astrocytic glutamate transporter disruption contributes to complement C1q-mediated microglial pruning of glutamatergic synapses
Kim et al. Loss of IQSEC3 disrupts GABAergic synapse maintenance and decreases somatostatin expression in the hippocampus
Balu et al. A small-molecule TLR4 antagonist reduced neuroinflammation in female E4FAD mice
Du et al. Subthalamic nucleus deep brain stimulation protects neurons by activating autophagy via PP2A inactivation in a rat model of Parkinson's disease
Zanon et al. MHC I upregulation influences astroglial reaction and synaptic plasticity in the spinal cord after sciatic nerve transection
JP2009149524A (ja) アルツハイマー病の予防・治療剤
Sifonios et al. An enriched environment restores normal behavior while providing cytoskeletal restoration and synaptic changes in the hippocampus of rats exposed to an experimental model of depression
Yenkoyan et al. Neuroprotective action of proline-rich polypeptide-1 in β-amyloid induced neurodegeneration in rats
US20250339422A1 (en) Methods for enhancing cellular clearance of pathological molecules via activation of the cellular protein ykt6
WO2025137265A1 (fr) Procédés et compositions pour réduire la gliaptose : mort cellulaire neuronale induite par macroglie
Zhan et al. ADSC-derived exosomes provide neuroprotection in sepsis-associated encephalopathy by regulating hippocampal pyroptosis
CA2854225A1 (fr) Co-activation des voies mtor et stat3 pour promouvoir la survie et la regenerescence neuronales

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24841379

Country of ref document: EP

Kind code of ref document: A1