Improvement of blood brain barrier integrity

Abstract

The present disclosure relates generally to the use of agents, such as oligonucleotides, to improve, increase, restore or retain the integrity of the blood brain barrier; and / or to reduce the deposition or accumulation of and / or promote the clearance of, amyloid-β. The disclosure also related to agents, such as oligonucleotides, that increase, promote or restore the level or amount of VE-cadherin in a cell, and are thus useful in the methods of the present disclosure. The present disclosure also relates to the use of such agents to treat diseases and conditions associated with damage to the blood brain barrier, including diseases and conditions associated with neuroinflammation and neurodegeneration, as well as diseases and conditions associated with amyloid-β deposition, including Alzheimer's disease (AD), Cerebral amyloid angiopathy (CAA), Lewy body dementia (LBD) and traumatic brain injury (TBI).

Description

IMPROVEMENT OF BLOOD BRAIN BARRIER INTEGRITYField of the Disclosure

[0001] The present disclosure relates generally to the use of agents, such as oligonucleotides, to improve, increase, restore or retain the integrity of the blood brain barrier; and / or to reduce the deposition or accumulation of and / or promote the clearance of, amyloid-g. The disclosure also related to agents, such as oligonucleotides, that increase, promote or restore the level or amount of VE-cadherin in a cell, and are thus useful in the methods of the present disclosure. The present disclosure also relates to the use of such agents to treat diseases and conditions associated with damage to the blood brain barrier, including diseases and conditions associated with neuroinflammation and neurodegeneration, as well as diseases and conditions associated with amyloid-3 deposition, including Alzheimer's disease (AD), Cerebral amyloid angiopathy (CAA), Lewy body dementia (LBD) and traumatic brain injury (TBI).Related Applications

[0002] This application claims priority to Australian Provisional Patent Application No. 2023903693 entitled " Improvement of blood brain barrier integrity" filed 16 November 2023, the contents of which are incorporated herein by reference in their entirety.Background of the Disclosure

[0003] The blood brain barrier (BBB) is the highly-specialized system of endothelial cells that plays a critical role in protecting the brain from infectious or toxic substances in the blood and controlling the influx and efflux of biological substances that are critical for brain metabolism and neuronal function.

[0004] Effective BBB function results from the complex and unique interaction between brain endothelial cells (ECs), pericytes, vascular smooth muscle cells, astrocytes, neurons, microglia, and a basement membrane, collectively referred to as the neurovascular unit (NVU). ECs of the BBB are morphologically and functionally distinguishable from peripheral ECs in several respects, including the absence of fenestrations, the lack of pinocytic activity, and the expression of active transport mechanisms to regulate the transport of essential molecules while preventing the passage of undesirable substances. ECs of the BBB are linked by tight junctions and adherens junctions that differ from their peripheral counterparts in the types and levels of molecules that form the junctions, and it is these junctions between the ECs that are predominantly responsible for so tightly controlling paracellular permeability. The critical components of the tight junctions in brain ECs include claudins-1, -3, -5 and -12, occludin, lipolysis-stimulated protein (LSR), and junctional adhesion molecule (JAM) -A, -B and -C, with claudin-5 being the most abundant and typically considered the most crucial for tight junction function. Zonula occludens (ZO)-l, -2 and -3 bind to motifs on intracellular domains of claudins and occludin and to the actin cytoskeleton, providing structural integrity to the tight junction. Adherins such as VE-cadherin and platelet endothelial cell adhesion molecule-1 (PECAM-1) are involved in the formation of adherensjunctions (see e.g. Knox et al., Molecular Psychiatry (2022) 27:2659-2673; Lochhead et al. (2020) Front. Physiol, Greene et al., (2019) Fluids Barriers CNS 16, 3). The specialized and distinct nature of the ECs in the BBB, combined with the activity of other cells and components of the NVU (such as the large coverage of pericytes wrapping around the ECs), results in a barrier with extremely low permeability and significantly higher transendothelial electrical resistance (TEER; ~ 1500-200 / cm2) than observed in ECs in the periphery (Stamatovic et al., Curr Neuropharmacol. 2008 Sep; 6(3): 179-192.).

[0005] Integrity of the BBB is essential for brain homeostasis. Conversely, loss of integrity of the BBB is associated with many serious diseases, including neurodegenerative and / or neuroinflammatory diseases, including dementias such as Alzheimer's disease (AD), as well as traumatic brain injury (TBI), and may play a role in the pathology of these diseases. For example, elevated blood pressure and / or pulse pressure can cause blood-brain barrier dysfunction and may be associated with dementia (Levin et al. 2020. Frontiers Neuroscience, art 669). However, the role of the BBB in the pathogenesis of diseases such as AD, cerebral amyloid angiopathy (CAA) and Lewy body dementia (LBD), and in particular the causative or correlative nature of the relationship and the relative importance of any one or more of the specialised molecules, cells or structures in the BBB in the development or progression of these diseases, is not well understood.

[0006] Amyloid-P and the plaques formed by and containing amyloid-p are associated with diseases such as AD, CAA, LBD and TBI, and are thought to be causative of at least some of the symptoms associated with those diseases.

[0007] AD is an age-related neurodegenerative brain disease characterized by the formation of extracellular amyloid-p plaques and intracellular neurofibrillary tangles in the brain, neuroinflammation, and neuronal and synaptic loss, resulting in memory loss, impaired cognition, and impaired reasoning or judgment. Alzheimer's disease is the most common form of dementia, and it is estimated that close to 7 million Americans may have the disease.

[0008] CAA is a type of cerebrovascular disorder characterized by the accumulation of amyloid-p within the leptomeninges and small to medium-sized cerebral blood vessels. Deposition of amyloid-p is thought to result in a weakening of the blood vessels, which may result in intracerebral hemorrhages (ICH). Patients with CAA can also present with amyloid-related imaging abnormalities (ARIA).

[0009] Despite decades of research and the development of a multitude of candidate therapeutics, few effective agents for restoring BBB integrity or preventing or reducing amyloid- 8 exist. There is therefore a need for new therapeutics for this purpose.Summary of the Disclosure

[0010] The present disclosure is predicated in part on the unexpected finding that oligonucleotides that inhibit the binding of miR-27a to VE-cadherin mRNA and thus increase or restore VE-cadherin levels are effective in restoring or retaining the integrity of the blood brainbarrier (BBB) and vasculature of the brain, reducing senescence in brain cells, and inhibiting and / or reducing amyloid-g deposition in the brain.

[0011] As described above, the BBB is a highly-specialized system of ECs that interact with pericytes, vascular smooth muscle cells, astrocytes, neurons, microglia, and a basement membrane to form the biological barrier at the blood to brain interface that effectively separates the brain from the rest of the body. The ECs of the BBB are quite distinct from ECs of the periphery in several respects, including the nature and properties of the junctions between the ECs. At least in part because of the types and / or expression levels of molecules that form the tight and adherens junctions, in particular the claudins such as claudin-5, the ECs of the BBB have reduced permeability and increased TEER compared to ECs in the periphery. The function of the BBB is further supported by other cells and structures, such a pericytes and astrocytes, resulting in an almost-impermeable barrier that is distinct from that seen in the vasculature of the periphery.

[0012] Despite VE-cadherin being just one molecule amongst many molecules and cells that function in the highly complex structure that is the BBB, the present inventors have unexpectedly demonstrated that targeting this one molecule to increase its expression is sufficient to restore or retain the integrity of the BBB and vasculature of the brain in mouse models of Alzheimer's disease. Moreover, as surprisingly demonstrated herein, targeting VE-cadherin with these molecules can inhibit and / or reduce senescence of brain cells and can also inhibit and / or reduce amyloid-g deposition, formation and / or accumulation. In some examples, the inhibition of amyloid-g deposition is within the vasculature of brain. In further examples, inhibition of amyloid- P deposition may be outside the vasculature, e.g. in the brain parenchyma.

[0013] Thus, provided herein are methods for improving, restoring or retaining the integrity of the BBB in a subject; methods for inhibiting and / or reducing amyloid-p deposition, accumulation and / or plaque formation in the brain of a subject; and methods for reducing or inhibiting senescence of brain cells in a subject. Accordingly, also provided are methods for the treatment of diseases and conditions associated with the loss of BBB integrity, such as neuroinflammatory diseases, neurodegenerative diseases, TBI, hypertension and / or elevated pulse pressure. Also provided are methods for the treatment of diseases and conditions associated with amyloid-p deposition, including AD, CAA, LBD and TBI.

[0014] In one aspect, provided is a method for improving, increasing, restoring or retaining the integrity of the blood brain barrier (BBB) in a subject, comprising administering to the subject an effective amount of an oligonucleotide comprising a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG or UCACAGU, to said RNA.

[0015] In some examples, the subject has a disease or condition associated with a loss of BBB integrity or has been determined to have a likelihood of developing a disease or conditionassociated with a loss of BBB integrity. In one embodiment, the disease or condition associated with a loss of BBB integrity is a neuroinflammatory disease or a neurodegenerative disease, e.g. dementia, multiple sclerosis (MS), motor neurone disease (MND), Parkinson's disease and Huntington's disease. The dementia may be selected from Alzheimer's disease, Lewy body dementia (LBD), Parkinson's disease dementia (PDD), vascular dementia and frontotemporal dementia (FTD). In another embodiment, the disease or condition associated with a loss of BBB integrity is traumatic brain injury (TBI). In some examples, the subject has dementia and administration of the oligonucleotide results in an improvement in cognition or a slowing in the rate of decline of cognition. In other examples, the condition associated with a loss of BBB integrity is hypertension or elevated pulse pressure.

[0016] Also provided is a method for reducing or inhibiting the deposition or accumulation of amyloid-p in the brain of a subject, or for increasing or promoting the clearance of amyloid-g in the brain of a subject, comprising administering to the subject an effective amount of an oligonucleotide comprising a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG or UCACAGU, to said RNA. In some examples, the subject has been diagnosed with Alzheimer's disease (AD) Cerebral amyloid angiopathy (CAA) or Lewy body dementia (LBD), or has been determined to have a likelihood of developing AD, CAA or LBD.

[0017] Also provided is a method for treating a subject with a disease or condition associated with a loss of BBB integrity, comprising administering to the subject an effective amount of an oligonucleotide comprising a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG or UCACAGU, to said RNA.

[0018] In some examples, the disease or condition associated with a loss of BBB integrity is a neuroinflammatory disease or a neurodegenerative disease, e.g. dementia (multiple sclerosis (MS), motor neurone disease (MND), Parkinson's disease and Huntington's disease. The dementia may be selected from Alzheimer's disease, Lewy body dementia (LBD), Parkinson's disease dementia (PDD), vascular dementia and frontotemporal dementia (FTD). In other examples, the condition associated with a loss of BBB integrity is traumatic brain injury (TBI), hypertension or elevated pulse pressure.

[0019] Also provided is a method of treating a subject with AD, CAA, LBD or TBI, or a subject likely to develop AD, CAA or LBD, comprising administering to the subject an effective amount of an oligonucleotide comprising a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG or UCACAGU, to said RNA.

[0020] In the methods described above and herein, the oligonucleotide may comprise a contiguous sequence complementary to at least 8 contiguous bases of an RIMA sequence comprising SEQ ID NO: 2, or SEQ ID NO: 2 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG or UCACAGU, to said RNA. In some examples, the miR- 27a miRNA is hsa-miR-27a comprising the nucleotide sequence set forth in SEQ ID NO: 13.

[0021] In some embodiments, the oligonucleotide comprises a contiguous sequence complementary to a sequence of at least or about 7 bases, at least or about 8 bases, at least or about 9 bases, at least or about 10 bases, at least or about 11 bases, at least or about 12 bases, at least or about 13 bases, at least or about 14 bases, at least or about 15 bases, at least or about 16 bases, at least or about 17 bases, at least or about 18 bases, at least or about 19 bases, at least or about 20 bases, at least or about 22 bases, at least or about 25 bases, at least or about 30 bases, or at least or about 35 bases of SEQ ID NO: 2, or SEQ ID NO: 2 comprising 1, 2 or 3 substitutions.

[0022] In one example, the oligonucleotide binds to positions 22-27 of SEQ ID NO: 2, In some examples, base pairing between the oligonucleotide and SEQ ID NO: 2 includes positions 8-28, 8-27, 9-27, 10-27, 11-27, 12-27, 13-27, 14-27, 15-27, 16-27, 17-27, 18-27, 19-27, 20- 27, 21-27, 9-28, 10-28, 11-28, 12-28, 13-28, 14-28, 15-28, 16-28, 17-28, 18-28, 19-28, 20-28 or 21-28 of SEQ ID NO: 2.

[0023] In particular examples, the oligonucleotide comprises the sequence set forth in any one of SEQ ID Nos: 3 to 6.

[0024] The oligonucleotide may also comprise one or more modified nucleobases, e.g. a LNA nucleobase, a UNA nucleobase or a 21O-methyl nucleobase. In particular examples, the oligonucleotide comprises a sequence set forth in any one of SEQ ID NOs: 7 to 11.

[0025] In some examples, the treatment results in a reduction or inhibition of the deposition or accumulation of amyloid-p in the brain of the subject; and / or results in the promotion or increase in the clearance of amyloid-p in the brain of the subject, e.g. in the vasculature of the brain and / or the paranchyma of the brain. In some examples, the reduction or inhibition of the deposition or accumulation of amyloid-p, and / or the promotion or increase in the clearance of amyloid-p, is in the leptomeninges vessels of the brain. In further examples, the amyloid-p is in the form of amyloid- 8 plaques.

[0026] The methods of the disclosure may further comprise administering a further therapy, such as a therapy that comprises another active agent, a device, physical therapy, cognitive therapy, and / or occupational therapy. In a particular example, the further therapy comprises an active agent or device that reduces blood pressure (e.g. treats hypertension) and / or pulse pressure.

[0027] In particular examples of the methods, administration of the oligonucleotide and optionally the further therapy results in an improvement in cognition or a slowing in the rate of decline of cognition.

[0028] Also provided is a use of an oligonucleotide in the preparation of a medicament for reducing or inhibiting the deposition or accumulation of amyloid-g plaques in the brain of a subject, or increasing or promoting the clearance of amyloid-g plaques in the brain of a subject, wherein the oligonucleotide comprises a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG or UCACAGU, to said RNA. In some examples, the subject has been diagnosed with Alzheimer's disease (AD), Cerebral amyloid angiopathy (CAA), Lewy body dementia (LBD) or traumatic brain injury (TBI), or has been determined to have a likelihood of developing AD, CAA or LBD.

[0029] Further aspects related to the use of an oligonucleotide in the preparation of a medicament for treating a subject with Alzheimer's disease (AD), Cerebral amyloid angiopathy (CAA), Lewy body dementia (LBD) or traumatic brain injury (TBI), wherein the oligonucleotide comprises a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG or UCACAGU, to said RNA.

[0030] Still further aspects relate to the use of an oligonucleotide in the preparation of a medicament for restoring or retaining the integrity of the blood brain barrier (BBB) in a subject, wherein the oligonucleotide comprises a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG or UCACAGU, to said RNA. In some examples, the subject has a disease or condition associated with a loss of BBB integrity or has been determined to have a likelihood of developing a disease or condition associated with a loss of BBB integrity, e.g. a neuroinflammatory disease and / or a neurodegenerative disease, such as one selected from a dementia, multiple sclerosis (MS), motor neurone disease (MND), Parkinson's disease and Huntington's disease; or TBI, hypertension and / or elevated pulse pressure. In some examples, the dementia is selected from Alzheimer's disease, Lewy body dementia (LBD), Parkinson's disease dementia (PDD), vascular dementia and frontotemporal dementia (FTD).Brief Description of the Drawings

[0031] Embodiments of the disclosure are described herein, by way of non-limiting example only, with reference to the following drawings.

[0032] Figure 1 is a photograph of brain tissue sections from 4 month old APPswe / PSldE9 administered CD5-2 or a control blockmir, and biotin, prior to euthanasia. Biotin leak (grey, red boxes) was detected in mice that received the control, while mice that received CD5-2 showed little or no biotin leak.

[0033] Figure 2 is a photograph of brain tissue sections from 7 month old APPswe / PSldE9 administered CD5-2 or a control blockmir prior to euthanasia. Albumin leak was observed in mice that received the control, while mice that received CD5-2 showed comparably reduced albumin leak.

[0034] Figure 3 is a photograph of brain tissue sections from 6 month old APPswe / PSldE9 administered CD5-2 or a control blockmir prior to euthanasia. Significant amyloid-3 deposition was observed in the leptomeningeal vessels (LV) and in the brain parenchyma of mice that received the control blockmir, while mice that received CD5-2 showed significantly reduced amyloid-3 deposition particularly in the LV.

[0035] Figure 4 is a photograph of brain tissue sections from wild type mice or from 6 month old APPswe / PSldE9 mice administered CD5-2 or a control blockmir prior to euthanasia. Albumin leak (green, arrows) and reduced VE-cadherin expression (red) was observed in APPswe / PSldE9 mice that received the control blockmir. Administration of CD5-2 to APPswe / PSldE9 mice restored VE-cadherin expression and reduced albumin leak to that observed in WT mice.

[0036] Figure 5 shows activity of blockmirs CD5-2, CD5-4 and CD5-9, specifically VE- cadherin expression (relative to GADPH) in endothelial cell isolates as performed by Western blot.A. Relative VE-cadherin expression in cells transfected with control blockmir, CD5-2 and CD5-4.B. Relative VE-cadherin expression in cells transfected with control blockmir, CD5-2 and CD5-9.C. Schematic of the structure of the blockmirs.

[0037] Figure 6 is a graphical representation of VE-cadherin and Claudin-5 expression in endothelial cell junctions of leptomeninges of APPswe / PSldE9 mice administered CD5-2 or a control blockmir. Brain tissue sections were stained and analysed by confocal microscopy, with mean intensity per leptomeningeal area shown. Mean from n=2-3 mice / gp from 2 separate experiments. A. VE-Cadherin (VEC). B. Claudin-5.

[0038] Figure 7 shows albumin leak from the leptomeninges of 6 month old APPswe / PSldE9 mice administered CD5-2 or a control blockmir. A single mouse received scrambled blockmir control and 2 littermates received CD5-2. Brain tissue sections were stained for CD31 (red) or albumin (green) and analysed by confocal microscopy. A. Representative section from mouse administered scrambled control blockmir. Arrow showing albumin leak. B. Representative section from mouse administered CD5-2. Arrow showing albumin leak. C. Graphical representation showing number of albumin leaks per cortical vessel area.

[0039] Figure 8 is a graphical representation of vessel density in APPswe / PSldE9 mice administered CD5-2 or a control blockmir. Brain tissue sections were stained for CD31 and analysed by confocal microscopy, and the area of CD31-positive vessels measured. Mean from n=2-3 mice / gp from 2 separate experiments.

[0040] Figure 9 shows amyloid deposition in leptomeninges and parenchyma of APPswe / PSldE9 mice administered CD5-2 or a control scrambled blockmir. A single mouse received scrambled blockmir control and 2 littermates received CD5-2. Brain tissue sections were stained for beta amyloid (green) and with DAPI (blue) and analysed by confocal microscopy. A. Representative stained sections from a mouse administered scrambled control blockmir and a mouse administered CD5-2. White dashed lines show where the leptomeningeal and penetrating vessels are located. B. Pairwise comparison of amyloid count in the cortex of mice (shown as total number of amyloid deposited relative to perimeter size of the cortex region). N=4 per group.

[0041] Figure 10 shows senescence in brain cells of 7 month old APPswe / PSldE9 mice administered CD5-2 or a control blockmir prior. Brain tissue sections were stained for p21 (red), laminBl (green) and with DAPI (blue) and analysed by confocal microscopy. A. Representative section from a mouse administered CD5-2 and a mouse administered the scrambled control blockmir. DAPI and p21 staining. B. Representative stained sections as in A). p21 and LaminBl staining. C. Magnified view of dashed box (a). D. Magnified view of dashed box (b). Senescent leptomeningeal cells and neurons had high p21 expression localized to nuclei (DAPI) and loss of LaminBl nuclear envelope. White arrows show leptomeningeal vascular-associated cells that are senescent. Yellow arrows show a neuronal population that are senescent.

[0042] Some figures and text contain color representations or entities. Color illustrations are available from the Applicant upon request or from an appropriate Patent Office. A fee may be imposed if obtained from a Patent Office.Detailed Description

[0043] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the disclosure belongs. All patents, patent applications, published applications and publications, databases, websites and other published materials referred to throughout the entire disclosure, unless noted otherwise, are incorporated by reference in their entirety. In the event that there is a plurality of definitions for terms, those in this section prevail. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference to the identifier evidences the availability and public dissemination of such information.

[0044] As used herein, the singular forms "a", "an" and "the" also include plural aspects (i.e. at least one or more than one) unless the context clearly dictates otherwise. Thus, for example, reference to "a polypeptide" includes a single polypeptide, as well as two or more polypeptides.

[0045] In the context of this specification, the term "about," is understood to refer to a range of numbers that a person of skill in the art would consider equivalent to the recited value in the context of achieving the same function or result.

[0046] Throughout this specification and the claims that follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0047] As used herein the term "oligonucleotide" refers to a single-stranded sequence of ribonucleotide or deoxyribonucleotide bases, known analogues of natural nucleotides, or mixtures thereof. An "oligonucleotide" comprises a nucleic-acid based molecule including DNA, RNA, PNA, LNA, UNA or any combination thereof. An oligonucleotide that predominantly comprises ribonucleotide bases, natural or non-natural, may be referred to as an RNA oligonucleotide. Oligonucleotides are typically short (for example less than 50 nucleotides in length) sequences that may be prepared by any suitable method, including, for example, direct chemical synthesis or cloning and restriction of appropriate sequences.

[0048] "Antisense oligonucleotides" are oligonucleotides complementary to a specific DNA or RNA sequence. Typically in the context of the present invention an antisense oligonucleotide is an RNA oligonucleotide complementary to a specific mRNA or miRNA. The antisense oligonucleotide binds to and silences or represses, partially or fully, the activity of its complementary miRNA. Not all bases in an antisense oligonucleotide need be complementary to the 'target' or miRNA sequence; the oligonucleotide need only contain sufficient complementary bases to enable the oligonucleotide to recognise the target. An oligonucleotide may also include additional bases. The antisense oligonucleotide sequence may be an unmodified ribonucleotide sequence or may be chemically modified or conjugated by a variety of means as described herein.

[0049] The term "polynucleotide" as used herein refers to a single- or double- stranded polymer of deoxyribonucleotide, ribonucleotide bases or known analogues of natural nucleotides, or mixtures thereof. A "polynucleotide" comprises a nucleic-acid based molecule including DNA, RNA, PNA, LNA, UNA or any combination thereof. The term includes reference to the specified sequence as well as to the sequence complimentary thereto, unless otherwise indicated. Polynucleotides may be chemically modified by a variety of means known to those skilled in the art. Thus a "polynucleotide" comprises a nucleic-acid based molecule including DNA, RNA, PNA, LNA, UNA or any combination thereof.

[0050] As used herein in relation to oligonucleotides and polynucleotides, the term "nucleotide" refers to a single nucleobase or monomer unit within the oligonucleotide or polynucleotide. The terms "nucleotide" and "monomer" may be used interchangeably herein. The nucleobase may be part of a DNA, RNA, INA, LNA, UNA (or combination of any two or morethereof) oligonucleotide or polynucleotide. In some embodiments, the nucleobase may be a universal base. Modified nucleobases are also contemplated by the present invention, as described hereinbelow.

[0051] The term "variant" as used herein refers to substantially similar sequences. Generally, polypeptide sequence variants also possess qualitative biological activity in common, such as receptor binding activity. Further, these polypeptide sequence variants may share at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity. The term "sequence identity" or "percentage of sequence identity" may be determined by comparing two optimally aligned sequences or subsequences over a comparison window or span, wherein the portion of the polynucleotide sequence in the comparison window may optionally comprise additions or deletions (i.e., gaps) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences.

[0052] The term "complementary" as used herein refers to the ability of two single-stranded nucleotide sequences to base pair, typically according to the Watson-Crick base pairing rules, that is, between G and C and between A and T or U. In some embodiments, G also pairs to U and vice versa to form a so-called wobble base pair. In another embodiment, the base inosine (I) may be included within an oligonucleotide of the invention. I base pairs to A, C and U. In still another embodiment, universal bases may be used. Universal bases can typically base pair to G, C, A, U and T. Often universal bases do not form hydrogen bonds with the opposing base on the other strand. In still another embodiment, a complementary sequence refers to a contiguous sequence exclusively of Watson-Crick base pairs. For two nucleotide molecules to be complementary they need not display 100% complementarity across the base pairing regions, but rather there must be sufficient complementarity to enable base pairing to occur. Thus a degree of mismatching between the sequences may be tolerated and the sequences may still be complementary. As used herein, the term "capable of base pairing with" is used interchangeably with "complementary to".

[0053] The term "substitution" as used herein refers to a nucleobase at a particular position within an oligonucleotide or polynucleotide having been substituted for another nucleobase. The substitution may be, for example, because of the presence of a single nucleotide polymorphism in the target RIMA. The term substitution also encompasses deletions of nucleobases and additions of nucleobases.

[0054] The term "blockmir" as used herein refers to a steric blocking oligonucleotide that binds to an RNA target, thereby blocking the ability of one or more miRNA species from binding to, and affecting the activity of, said target. Blockmirs are constructed so as to be incapable of recruiting cellular RNAi machinery or RNase H. RNAi machinery refers to the cellular componentsnecessary for the activity of siRNAs and miRNAs or for the RNAi pathway. A major component of the RNAi machinery is the RNA induced silencing complex (the RISC complex). Blockmirs are described, for example, in WO 2008 / 061537, WO 2012 / 069059 and WO 2014 / 053014, the disclosures of which are incorporated herein by reference.

[0055] In the context of this specification, the term "activity" as it pertains to a polynucleotide (e.g. a DNA, mRNA or miRNA), protein or polypeptide means any one or more cellular function, action, effect or influence exerted by the polynucleotide, protein or polypeptide. For example, in the context of a mRNA, activity will typically refer to expression of the mRNA, i.e. translation into a protein or peptide. Thus, regulation of the activity of a target mRNA by an oligonucleotide as described herein may include degradation of the mRNA and / or translational regulation. Regulation of mRNA activity may also include affecting intracellular transport of the mRNA. In the context of an oligonucleotide of the invention, "activity" typically means the ability of the oligonucleotide to inhibit the interaction between miR-27a and VE-cadherin, thereby increasing VE-Cadherin expression.

[0056] The term "inhibiting" and variations thereof such as "inhibition" and "inhibits" as used herein do not necessarily imply the complete inhibition of the specified event, activity or function. Rather, the inhibition may be to an extent, and / or for a time, sufficient to produce the desired effect. Inhibition may be prevention, retardation, reduction or otherwise hindrance of the event, activity or function. Such inhibition may be in magnitude and / or be temporal in nature. In particular contexts, the terms "inhibit", "reduce" and "prevent", and variations thereof may be used interchangeably. The inhibition of amyloid-p plaque formation or deposition or accumulation by oligonucleotides of the invention may be direct or indirect and may be in magnitude and / or be temporal in nature.

[0057] The terms "promoting" and "inducing", and variations thereof such as "promotion" and "inducement", as used herein do not necessarily imply the complete promotion or inducement of the specified event, activity or function. Rather, the promotion or inducement may be to an extent, and / or for a time, sufficient to produce the desired effect. The promotion or inducement of amyloid-p plaque clearance by oligonucleotides of the invention may be direct or indirect and may be in magnitude and / or be temporal in nature.

[0058] As used herein the term "effective amount" includes within its meaning a non-toxic but sufficient amount or dose of an agent or compound to provide the desired effect. The exact amount or dose required will vary from subject to subject depending on factors such as the species being treated, the age and general condition of the subject, the severity of the condition being treated, the particular agent being administered and the mode of administration and so forth. Thus, it is not possible to specify an exact "effective amount". However, for any given case, anappropriate "effective amount" may be determined by one of ordinary skill in the art using only routine experimentation.

[0059] As used herein the terms "treating", "treatment", "preventing" and "prevention" refer to any and all uses which remedy a condition or symptoms, prevent the establishment of a condition or disease, or otherwise prevent, hinder, retard, or reverse the progression of a condition or disease or other undesirable symptoms in any way whatsoever. Thus the terms "treating" and "preventing" and the like are to be considered in their broadest context. For example, treatment does not necessarily imply that a patient is treated until total recovery. In conditions which display or a characterized by multiple symptoms, the treatment or prevention need not necessarily remedy, prevent, hinder, retard, or reverse all of said symptoms, but may prevent, hinder, retard, or reverse one or more of said symptoms. In the context of some disorders, methods of the present invention involve "treating" the disorder in terms of reducing or ameliorating the occurrence of a highly undesirable event associated with the disorder or an irreversible outcome of the progression of the disorder but may not of itself prevent the initial occurrence of the event or outcome. Accordingly, treatment includes amelioration of the symptoms of a particular disorder or preventing or otherwise reducing the risk of developing a particular disorder. Indicia of successful "treatment", includes any objective or subjective parameter such as abatement; remission; improvement in or a slowing of decline in memory; condition more tolerable to the patient; slowing in the rate of degeneration or decline or worsening of the illness; making the final point of worsening less debilitating; or improving a subject's physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters; including the results of a physical examination, neurological examination, and / or psychiatric evaluations. In some examples of the present disclosure, where the disease being treated is a dementia, treatment may result in amelioration or reduction in cognitive decline (or the rate of cognitive decline) and / or an improvement in cognitive function.

[0060] As used herein, reference to a disease or condition associated with a loss of BBB integrity means a disease or condition that is characterized by (i.e. has evidence of) a loss of BBB integrity. Loss of BBB integrity is typically assessed by measuring BBB permeability. This can be done in animal models of the disease, using ex vivo analysis and / or in vivo in the clinical setting with patients suspected of having or being likely to develop a disease or condition associated with a loss of BBB integrity. Loss of BBB integrity may contribute to the pathology, clinical outcomes and / or symptoms of the disease, and may include neuroinflammation, neurodegradation (e.g. loss of neuron function), plaque deposition, cognitive decline or impairment, muscle weakness and / or mobility decline. Diseases or conditions associated with a loss of BBB integrity include neuroinflammatory diseases, neurodegenerative diseases, TBI, hypertension and / or elevated pulse pressure.

[0061] Reference to "improving, restoring or retaining the integrity of the blood brain barrier (BBB)" (or grammatical variations thereof) means improving, restoring or retaining the ability of the BBB to function as a barrier at the blood to brain interface, so as to, for example, preventmacromolecules from entering the brain. In some examples, BBB is improved by at least or about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or more compared to a reference (e.g. an earlier timepoint, or a comparative BBB). In some examples, BBB integrity is restored (e.g. to at least 80%, 85%, 90% or 95%) to that observed at an earlier timepoint or to a control or reference (e.g. a normal or healthy control or reference). In other examples, BBB is retained, i.e. is not lost or reduced. BBB integrity is typically assessed by measuring BBB permeability using methods well known in the art, such as those described herein, with permeability being inversely related to BBB integrity.

[0062] As used herein, reference to a disease or condition associated with amyloid-p deposition means a disease or condition that is characterized by (i.e. has evidence of) amyloid-p deposition. Typically, the amyloid-p deposition is in the brain (e.g. in the brain vasculature and / or parenchyma). Amyloid-P deposition can be assessed directly using various imaging techniques, or indirectly using markers, such as blood or CSF amyloid-p levels. Diseases or conditions associated with amyloid-p deposition include AD, CAA, LBD and TBI.

[0063] As used herein, "Alzheimer's disease" or AD refers to a disease characterized by progressive cognitive impairment. The symptoms of Alzheimer's disease typically worsen over time as the disease progresses, with the disease typically progressing through three stages: "mild" (an early-stage form of Alzheimer's disease), "moderate" (a middle-stage form), and "severe" (a late-stage form). In mild Alzheimer's disease, symptoms may include, for example, memory loss, losing or misplacing objects, trouble remembering names or recalling words, increased difficulty with planning or organizing, taking longer to complete normal daily tasks, and repeating questions. In moderate Alzheimer's disease, which is typically the longest stage of the disease for many patients, damage occurs in areas of the brain that control language, reasoning, sensory processing, and conscious thought. In this stage, symptoms may include, for example, forgetfulness of events or of one's one personal history, problems recognizing family and friends, inability to learn new information, difficulty carrying out multi-step tasks, impulsive behavior, changes in sleep patterns, hallucinations, delusions, and paranoia. In severe Alzheimer's disease, memory and cognitive skills continue to worsen, patients typically lose the ability to respond to their environment, carry on a conversation, and / or control movement, and patients require a high level of assistance with daily activities and personal care.

[0064] In some embodiments, a patient has "late onset" Alzheimer's disease, which refers to a form of Alzheimer's disease in which the patient exhibits clinical symptoms of the disease after about age 65. In some embodiments, a patient has "early onset" Alzheimer's disease, which refers to a form of Alzheimer's disease in which a patient exhibits the onset of clinical symptoms of the disease prior to the age of 65. In some embodiments, patients having early onset Alzheimer's disease exhibit the onset of clinical symptoms of the disease in their 30s, 40s, or 50s. In some embodiments, patients have familial Alzheimer's disease (FAD), which is a hereditary form of Alzheimer's disease caused by autosomal dominant mutations that affect APP processing. Reference to Alzheimer's disease also include early stage Alzheimer's disease.

[0065] "Cerebral amyloid angiopathy" or CAA is a type of cerebrovascular disorder characterized by the accumulation of amyloid-g within the leptomeninges and small to mediumsized cerebral blood vessels. Deposition of amyloid-p is thought to result in a weakening of the blood vessels, which may result in intracerebral hemorrhages (ICH). Subjects with CAA may also present with cognitive impairments, incidental microbleeds, hemosiderosis, inflammatory leukoencephalopathy, Alzheimer's disease, or transient neurological symptoms. While a definitive diagnosis of CAA can only be made through a postmortem examination of the brain, a "probable" CAA diagnosis is made during the subject's life by imaging or tissue sampling. For purposes of the present disclosure, diagnosis of probable CAA is diagnosis of CAA. Evidence of CAA includes lobar, cortical-subcortical hemorrhage, or cortical hemorrhage; pathological evidence of CAA; multiple hemorrhages restricted to the cortical, lobar, or cortical-subcortical regions; and / or single lobar, cortical, or cortical-subcortical hemorrhage and focal or disseminated superficial siderosis.

[0066] "Traumatic brain injury" or"TBI" refers to an injury to the brain caused by an external force. TBI can be classified based on severity (e.g. mild, moderate or severe), mechanism (closed or penetrating head injury), or other features (such as occurring in a specific location (e.g. focal) or over a widespread area (e.g. diffuse). TBI can lead to temporary or permanent impairment of cognitive, physical and psychosocial functions. TBI is not a single pathophysiological event but a complex disease process, with structural damage and functional deficits that are due to both primary and secondary injury mechanisms. The primary injury is the result of the immediate mechanical disruption of brain tissue that occurs at the time of exposure to the external force and includes contusion, damage to blood vessels (hemorrhage), and axonal shearing, in which the axons of neurons are stretched and torn. The secondary injury evolves over minutes to months to years after the primary injury, resulting from cascades of various metabolic, cellular and molecular events (including amyloid plaque formation) that ultimately lead to brain cell death, tissue damage and atrophy. TBI is considered a major risk factor for dementia, including AD.

[0067] As used herein, "APP" refers to "amyloid precursor protein." The protein encoded by the APP gene is a type I membrane protein having an El domain and E2 domain. Cleavage of the APP protein produces an amyloid beta (Ag) fragment and the APP intracellular domain (AICD). Human APP gene and protein sequences, including splice and isoform variants, are well known and set forth in, e.g., NCBI GenBank Accession Nos. AH005295.2 and NM_000484.3. In some embodiments, the APP gene or protein is a variant (e.g., polymorphic variant, splice variant, or truncated protein) having at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a naturally occurring APP gene or protein (such as any set forth above) or a fragment thereof.

[0068] The terms "amyloid-P" or "amyloid-p peptides" or AP may be used interchangeably and refer to peptides of between about 27 and 49 amino acids that result from enzymatic processing of APP. Amyloid-P populations are heterogeneous, although the 40 amino acid peptide (AP40) is the most abundant (about 80-90%), followed by the 42 amino acid peptide (A[342,about 5-10%). Longer forms of AP, and in particular AP42, are more hydrophobic and f i bri I logenic, and are the principal species deposited in the brain. In contrast, AP40 is more commonly observed in cerebral amyloid angiopathy (CAA), though AP42 also is generally present. "Amyloid-p plaque" refers to an insoluble aggregate of amyloid-p, and in particular of amyloid-p fibrils. Amyloid-p and amyloid-p plaques in the brain can be detected and measured by any known method, including staining of tissues with anti-AP antibodies and brain imaging techniques such as positron emission tomography (PET). In other cases, brain amyloid-p is detected indirectly by detecting markers of brain amyloid-p. Non-limiting examples include the detection of amyloid-p (and in particular AP40 and / or AP42) in cerebrospinal fluid (CSF) or plasma samples (see e.g. Li et al., Neurology. 2022 Feb 15; 98(7): e688-e699).

[0069] The term "subject" as used herein refers to mammals and includes humans, primates, livestock animals (e.g. sheep, pigs, cattle, horses, donkeys), laboratory test animals (eg. mice, rabbits, rats, guinea pigs), companion animals (eg. dogs, cats) and captive wild animals (eg. foxes, kangaroos, deer). Preferably, the mammal is human or a laboratory test animal. Even more preferably, the mammal is a human. "Subject" and "patient" may be used interchangeably herein.

[0070] It will be appreciated that the above described terms and associated definitions are used for the purpose of explanation only and are not intended to be limiting.Table 1. SequencesSingle underlining represents a locked nucleic acid (LNA) monomer; double underlining represents a 2' O-methyl RNA monomer; bold represents phosphonothioate; italics represents a UNA monomerOligonucleotides

[0071] Provided are oligonucleotides that are suitable for the uses and methods described herein. Such oligonucleotides include those that inhibit the interaction between the 3' untranslated region (3'UTR) of the VE-cadherin (CDH5) mRNA molecule comprising the sequence shown in SEQ ID NO: 1, and an inhibitory miRNA. The oligonucleotides bind the 3'UTR so as to inhibit the interaction between an miRNA comprising the seed sequence UCACAG or UCACAGU and the 3'UTR of VE-cadherin. In particular examples, the miRNA is miR-27a. The nucleotide sequence of mature human miR-27a (hsa-miR-27a) is provided in SEQ ID NO: 13. Also contemplated herein are variants of this miRNA. Variants include nucleotide sequences that are substantially similar to the sequence of miR-27a. For example, a variant miRNA may comprise a sequence displaying at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 13. By inhibiting the interaction between miR27a and VE- cadherin, the oligonucleotides act to increase the expression of VE-cadherin compared to expression levels seen in the absence of the oligonucleotide. In some examples, expression of VE-cadherin is increased by at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,100%, 200%, 300% or more (see e.g. International Patent Publication No. W02014053014 and the Examples below).

[0072] The oligonucleotides typically comprise a contiguous sequence complementary to a sequence selected from the group consisting of at least about 9 contiguous bases, at least about 10 contiguous bases, at least about 11 contiguous bases, at least about 12 contiguous bases, at least about 13 contiguous bases, at least about 14 contiguous bases, at least about 15 contiguous bases, at least about 16 contiguous bases, at least about 17 contiguous bases, at least about 18 contiguous bases, at least about 19 contiguous bases, at least about 20 contiguous bases, at least about 22 contiguous bases, at least about 25 contiguous bases, at least about 30 contiguous bases, and at least about 35 contiguous bases of the sequence set forth in SEQ ID NO: 2 or the sequence of SEQ ID NO: 2 comprising 1, 2 or 3 substitutions.

[0073] In an embodiment, the oligonucleotide may comprise a contiguous sequence complementary to a sequence selected from the group consisting of no more than 8 contiguous bases, no more than 9 contiguous bases, no more than 10 contiguous bases, no more than 11 contiguous bases, no more than 12 contiguous bases, no more than 13 contiguous bases, no more than 14 contiguous bases, no more than 15 contiguous bases, no more than 16 contiguous bases, no more than 17 contiguous bases, no more than 18 contiguous bases, no more than 19 contiguous bases, no more than 20 contiguous bases, no more than 22 contiguous bases, no more than 25 contiguous bases, no more than 30 contiguous bases, and no more than 35 contiguous bases of the sequence set forth in SEQ ID NO: 2 or the sequence of SEQ ID NO: 2 comprising 1, 2 or 3 substitutions.

[0074] In another embodiment, the oligonucleotide may comprise a contiguous sequence complementary to a sequence selected from the group consisting of 8 contiguous bases, 9 contiguous bases, 10 contiguous bases, 11 contiguous bases, 12 contiguous bases, 13 contiguous bases, 14 contiguous bases, 15 contiguous bases, 16 contiguous bases, 17 contiguous bases, 18 contiguous bases, 19 contiguous bases, 20 contiguous bases, 21 contiguous bases, 22 contiguous bases, 23 contiguous bases, 24 contiguous bases, 25 contiguous bases, 30 contiguous bases, and 35 contiguous bases of the sequence set forth in SEQ ID NO: 2 or the sequence of SEQ ID NO: 2 comprising 1, 2 or 3 substitutions.

[0075] Typically, the oligonucleotide binds to positions 22-27 of SEQ ID NO: 2, this region representing the complement of the seed sequence of miR-27a, being the target site for miR-27a binding to the 3'UTR of the VE-cadherin mRNA (the 'anti-seed' region). Base pairing between the oligonucleotide and SEQ ID NO: 2 may include positions 8-32, 8-31, 8-30, 8-29, 8-28, 8-27, 9- 32, 9-31, 9-30, 9-29, 9-28, 9-27, 10-32, 10-31, 10-30, 10-29, 10-28, 10-27, 11-32, 11-31, 11- 30, 11-29, 11-28, 11-27, 12-27, 13-27, 14-27, 15-27, 16-27, 17-27, 18-27, 19-27, 20-27, 21-27, 9-28, 10-28, 11-28, 12-28, 13-28, 14-28, 15-28, 16-28, 17-28, 18-28, 19-28, 20-28 or 21- 28 of SEQ ID NO: 2.

[0076] In one embodiment, base pairing between the oligonucleotide and the sequence of SEQ ID NO: 2 ends at position 27 of SEQ ID NO: 2. In other embodiments, base pairing may end at position 28, 29, 30, 31, 32 or 33 of SEQ ID NO: 2. In another embodiment, base pairing between the oligonucleotide and the sequence of SEQ ID NO: 2 begins at position 22 of SEQ ID NO: 2. In other embodiments, base pairing may start at position 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6 or 5 of SEQ ID NO: 2.

[0077] Those skilled in the art will appreciate that the oligonucleotides may be of any suitable length depending on the precise function or use of the oligonucleotide. In some examples, the oligonucleotide is no more than 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75 or 80 bases in length. Typically, the oligonucleotides are between 8 and 25 bases in lengths. Even more typically, the oligonucleotides are between 10 and 20 bases in length. In some examples, the oligonucleotides are at least 8 bases in length and no more than 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 bases in length; are at least 9 bases in length and no more than 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 bases in length; are at least 10 bases in length and no more than 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 bases in length; are at least 11 bases in length and no more than 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 bases in length; or are at least 12 bases in length and no more than 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 bases in length.

[0078] For strong binding to its target RNA, the length of the oligonucleotide may be increased. In some cases, delivery into cells may be improved may using shorter oligonucleotides. Further, in other cases, the position of the oligonucleotide respective to the anti-seed sequence of the target RNA may be adjusted. For example, the position of bases complementary to position 22-27 of the target RNA of SEQ ID NO: 2 may be adjusted such that they are placed for example at the 5'end of the oligonucleotide, at the 3'end of the oligonucleotide or in or towards the middle of the oligonucleotide. Typically, the position of bases complementary to positions 22-27 are placed in the oligonucleotide such that they start at position 1, position 2, position 3, position 4, position 5 or position 6, or at a position upstream of position 2, position 3, position 4, position 5 or position 6 or at a position downstream of position 1, position 2, position 3, position 4, position 5 or position 6, wherein the positions are counted from the 5'end of the oligonucleotide.

[0079] In some embodiments, the target RNA sequence, for example the sequence of SEQ ID NO: 2 may comprise 1, 2 or 3 substitutions. Alternatively, the sequence may comprise no substitutions. Where substitutions are present, these may be located in the region of complementarity between the oligonucleotide and the target RNA. Substitutions may be single nucleotide polymorphisms (SNPs) that may enhance or decrease miRNA regulation of the giventarget RNA. An SNP may create a new miRNA target site so as to cause aberrant miRNA regulation of the given target RNA. RNA editing may also give rise to substitutions.

[0080] The oligonucleotides may be capable of activating RNase H. RNase H cleaves the RNA part of a RNA-DNA duplex and the structural requirements for RNase H activation are well- known to the skilled addressee. Similarly, oligonucleotides of the invention may be capable of recruiting the cellular RNAi machinery and directing the RNAi machinery to the target RNA. This may result in cleavage of the target RNA or translational repression of the target RNA.

[0081] In particular embodiments of the present invention, the oligonucleotides can neither recruit the RNAi machinery nor RNase H. Thus typically, oligonucleotides of the invention are capable of blocking the activity of the RNAi machinery at a particular target RNA. The oligonucleotides may do so by sequestering the target sequence (the miRNA binding site) of the target RNA, such that the RNAi machinery will not recognize the target sequence. Oligonucleotides of the invention with this activity may also be referred to as Blockmirs, because they block the regulatory activity of a given miRNA at a particular miRNA binding site in target RNA. To achieve the ability to prevent recruitment or activation of RNase H by oligonucleotides of the invention, the oligonucleotides typically do not comprise 5 or more contiguous DNA nucleobases.

[0082] The oligonucleotides may comprise a variety of sequence and structural modifications, depending on the use and function of the oligonucleotide, as will be described further below. Those skilled in the art will appreciate that the sequence and structural modifications described herein are exemplary only, and the scope of the present invention should not be limited by reference to those modifications, but rather additional modifications known to those skilled in the art may also be employed provided the oligonucleotide retains the desired function or activity.

[0083] By way of example only, the oligonucleotide sequence may be modified by the addition of one or more phosphorothioate (for example phosphoromonothioate or phosphorodithioate) linkages between residues in the sequence, or the inclusion of one or morpholine rings into the backbone. Alternative non-phosphate linkages between residues include phosphonate, hydroxlamine, hydroxylhydrazinyl, amide and carbamate linkages, methylphosphonates, phosphorothiolates, phosphoramidates or boron derivatives. The nucleotide residues present in the oligonucleotide may be naturally occurring nucleotides or may be modified nucleotides. Suitable modified nucleotides include 2'-O-methyl nucleotides, 2'-0- flouro nucleotides, 2'-O-methoxyethyl nucleotides, universal nucleobases such as 5-nitro-indole; LNA, UNA, PNA and INA nucleobases, 2'-deoxy-2'-fluoro-arabinonucleic acid (FANA) and arabinonucleic acid (ANA). The ribose sugar moiety that occurs naturally in ribonucleosides may be replaced, for example with a hexose sugar, polycyclic heteroalkyl ring, or cyclohexenyl group.Alternatively, or in addition, the oligonucleotide sequence may be conjugated to one or more suitable chemical moieties at one or both ends. For example, the oligonucleotide may be conjugated to cholesterol via a suitable linkage such as a hydroxyprolinol linkage at the 3' end. As a further example, the oligonucleotide may be conjugated to N-acetylgalactosamine (GalNAc).

[0084] Particular modifications of interest include those that increase the affinity of the oligonucleotide for complementary sequences, i.e. increase the melting temperature of the oligonucleotide base paired to a complementary sequence, or increase the biostability of the oligonucleotide. Such modifications include 2'-O-flouro, 2'-O-methyl, 2'-O-methoxyethyl groups. The use of LNA, UNA, PNA and INA monomers are also typically employed. For shorter oligonucleotides, typically a higher percentage of affinity increasing modifications are present. If the oligonucleotide is less than 12 or 10 nucleobases in length, it may be composed entirely of affinity increasing units, e.g. LNA monomers, UNA monomers or 2'-O-methyl RNA nucleobases.

[0085] In particular embodiments, the fraction of monomers in an oligonucleotide modified at either the base or sugar relatively to the monomers not modified at either the base or sugar may be less than 99%, less than 95%, less than 90%, less than 85 %, less than 80 %, less than 75%, less than 70%, less than 65%, less than 60%, less than 50 %, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 1%, more than 99%, more than 95%, more than 90%, more than 85 %, more than 75%, more than 70%, more than 65%, more than 60%, more than 50 %, more than 45%, more than 40%, more than 35%, more than 30%, more than 25%, more than 20%, more than 15%, more than 10%, and more than 5% or more than 1%.

[0086] Lipids and / or peptides may also be conjugated to the oligonucleotides. Such conjugation may both improve bioavailability and prevent the oligonucleotide from activating RNase H and / or recruiting the RNAi machinery. Conjugation of larger bulkier moieties is typically done at the central part of the oligonucleotide, e.g. at any of the most central 5 monomers. Alternatively, at one of the bases complementary to one of position 1-6 of SEQ ID NO: 1 or one of position 22-27 of SEQ ID NO: 2. In yet another embodiment, the moiety may be conjugated at the 5'end or the 3'end of the oligonucleotide. One exemplary hydrophobic moiety is a cholesterol moiety that may be conjugated to the oligonucleotide preventing the oligonucleotide from recruiting the RNAi machinery and improving bioavailability of the oligonucleotide. For example, the cholesterol moiety may be conjugated to one or more of the nucleobases complementary to positions 22-27 of the sequence of SEQ ID NO: 2, at the 3'end of the oligonucleotide, or at the 5'end of the oligonucleotide.

[0087] Different modifications may be placed at different positions within the oligonucleotide to prevent the oligonucleotide from activating RNase H and / or being capable of recruiting the RNAi machinery.

[0088] In a particular embodiment, phosphorothioate internucleotide linkages may connect the monomers in an oligonucleotide to improve the biostability of the oligonucleotide. All linkages of the oligonucleotide may be phosphorothioate linkages. In another embodiment, the fraction of phosphorothioate linkages may be less than 95%, less than 90%, less than 85 %, less than 80%, less than 75%, less than 70%, less than 65%, less than 60%, less than 50 %, more than 95%, more than 90%, more than 85 %, more than 80%, more than 75%, more than 70%, more than 65%, more than 60% and more than 50%. In some examples, all nucleobases are connected by phosphorothiate linkage except the 3' nucleobase.

[0089] In an embodiment, the oligonucleotide may not comprise any RNA nucleobases. This may assist in preventing the oligonucleotide from being capable of recruiting the RNAi machinery increasing biostability of the oligonucleotide. For example, the oligonucleotide may consist of LN A and DNA nucleobases and these may be connected by phosphorothioate linkages as outlined above. In alternative embodiments, the oligonucleotide does not comprise any DNA nucleobases. In alternative embodiments, the oligonucleotide does not comprise any morpholino and / or LNA nucleobases.

[0090] In an embodiment, the oligonucleotide may comprise a mix of DNA nucleobases and RNA nucleobases to prevent the oligonucleotide from activating RNase H and prevent the oligonucleotide from recruiting the RNAi machinery. For example, DNA and RNA nucleobases may be alternated along the length of the oligonucleotide, or alternatively one or more DNA nucleobases may be located adjacent one another and one or more RNA nucleobases may be located adjacent one another.

[0091] In another particular embodiment, the oligonucleotide comprises a mix of LNA monomers and 2'-O-methyl RNA nucleobases. As above, LNA and 2'-O-methyl RNA nucleobases may be alternated along the length of the oligonucleotide, or alternatively one or more LNA nucleobases may be located adjacent one another and one or more 2'-O-methyl RNA nucleobases may be located adjacent one another.

[0092] In some embodiments, the number of nucleobases present in an oligonucleotide that increase the affinity of the oligonucleotide for complementary sequences is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, or at least 22 nucleobases. In some embodiments, the number of nucleobases present in a oligonucleotide that increase the affinity of the oligonucleotide for complementary sequences is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and 22 nucleobases.

[0093] In particular embodiments, the nucleobases that increase the affinity of the oligonucleotide for complementary sequences may be located at the flanks of the oligonucleotide, i.e. at or near either or both of the 5' and 3' ends of the oligonucleotide, or may be located at or near the centre of the oligonucleotide. The nucleobases that increase the affinity of the oligonucleotide for complementary sequences may also be distributed evenly across the length of the oligonucleotide.

[0094] Table 2 sets out exemplary oligonucleotide sequences. Thus, provided herein is an oligonucleotide that comprises a sequence set forth SEQ ID N0:3, 4, 5 or 6, is less than 50 nucleotides in length and optionally further comprises at least one modified nucleobase, such as an LNA nucleobase and / or a 2' O-methyl nucleobase. Such oligonucleotides can inhibit binding of miR-27a to a RNA molecule comprising SEQ ID NO: 1, 2, 14 or 15. In another example, the oligonucleotide comprises a sequence set forth in SEQ ID N0:7, is less than 50 nucleotides in length (e.g. less than 40, 35, 30, 35 or 30 nucleotides), and can inhibit binding of miR-27a to a RNA molecule comprising SEQ ID NO: 1, 2, 14 or 15. In another example, the oligonucleotide comprises a sequence set forth in SEQ ID NO:8, is less than 50 nucleotides in length (e.g. less than 40, 35, 30, 35 or 30 nucleotides), and can inhibit binding of miR-27a to a RNA molecule comprising SEQ ID NO: 1, 2, 14 or 15. In another example, the oligonucleotide comprises a sequence set forth in SEQ ID NO:9, is less than 50 nucleotides in length (e.g. less than 40, 35, 30, 35 or 30 nucleotides), and can inhibit binding of miR-27a to a RNA molecule comprising SEQ ID NO: 1, 2, 14 or 15. In another example, the oligonucleotide comprises a sequence set forth in SEQ ID NO: 10, is less than 50 nucleotides in length (e.g. less than 40, 35, 30, 35 or 30 nucleotides), and can inhibit binding of miR-27a to a RNA molecule comprising SEQ ID NO: 1, 2, 14 or 15. In another example, the oligonucleotide comprises a sequence set forth in SEQ ID NO: 11, is less than 50 nucleotides in length (e.g. less than 40, 35, 30, 35 or 30 nucleotides), and can inhibit binding of miR-27a to a RNA molecule comprising SEQ ID NO: 1, 2, 14 or 15. The aforementioned oligonucleotides can increase expression and / or activity of VE-cadherin in a cell and reduce vascular leak or permeability. Thus, in methods that comprise contacting the cell with an oligonucleotide described above (e.g. in methods that comprise administering the oligonucleotide to a subject), the expression and / or activity of VE-cadherin is increased and / or vascular leak or permeability is decreased (such as compared to when the cell is not contacted with the oligonucleotide or the subject is not administered the oligonucleotide). By virtue of their ability to increase VE-cadherin levels and reduce vascular leak or permeability, the aforementioned oligonucleotides are suitable for use in methods for the treatment of associated diseases and conditions, including but not limited to those described herein, those described in WQ2014053014 (e.g. oedema, cardiovascular disease, myocardial infarction, peripheral vascular disease, ischaemia, stroke, cancer, atherosclerosis, psoriasis, diabetes, autoimmune diseases such as rheumatoid arthritis, thrombocytopenia, altitude sickness, barotrauma, iatrogenic disorders, bacterial infections, viral infections, and ocular conditions associated with vascular leak such as non-proliferative and proliferative retinopathies, macular oedema, glaucoma and macular degeneration).Table 2. Oligonucleotide sequencesSingle underlining represents an LNA monomer; double underlining represents a 2' O-methyl RNA monomer; bold represents a UNA monomerCompositions

[0095] Oligonucleotides may formulated in the form of pharmaceutical compositions, which compositions may comprise one or more pharmaceutically acceptable carriers, excipients or diluents. Such compositions may be administered in any convenient or suitable route such as by parenteral (e.g. subcutaneous, intraarterial, intravenous, intramuscular), oral (including sublingual), nasal or topical routes. In circumstances where it is required that appropriate concentrations of the oligonucleotide are delivered directly to the site in the body to be treated, administration may be regional rather than systemic. Regional administration provides the capability of delivering very high local concentrations of the oligonucleotide to the required site and thus is suitable for achieving the desired therapeutic or preventative effect whilst avoiding exposure of other organs of the body to the compound and thereby potentially reducing side effects.

[0096] Oligonucleotides of the invention may be packaged and delivered in suitable delivery vehicles which may serve to target or deliver the oligonucleotide or further protect or stabilize the oligonucleotide. By way of example, the delivery vehicle may comprise liposomes or other similar compositions such as micelles (e.g. polymeric micelles), lipoprotein-based drug carriers, microparticles, nanoparticles, liposome nanoparticles (LNPs), lipid nanoparticles or dendrimers.

[0097] Liposomes may be derived from phospholipids or other lipid substances, and are formed by mono- or multi-lamellar hydrated liquid crystals dispersed in aqueous medium. Specific examples of liposomes used in administering or delivering a composition to target cells are DODMA, synthetic cholesterol, DSPC, PEG-cDMA, DLinDMA, or any other non-toxic, physiologically acceptable and metabolisable lipid capable of forming liposomes. The compositions in liposome form may contain stabilisers, preservatives and / or excipients. Methods for preparing liposomes are well known in the art, for example see Methods in Cell Biology, VolumeXIV, Academic Press, New York, N.Y. (1976), p. 33 ff., the contents of which are incorporated herein by reference. Biodegradable microparticles or nanoparticles formed from, for example, polylactide (PLA), polylactide-co-glycolide (PLGA), and epsilon-caprolactone (e-caprolactone) may be used.

[0098] Other means of packaging and / or delivering oligonucleotides, and optionally one or more additional agents, in order to facilitate delivery to the brain will also be well known to those skilled in the art. By way of example only, delivery platforms may include RNA-lipoplex technologies comprising cationic lipids, fusogenic or stabilising co-lipids, and PEGylated lipids.

[0099] Examples of pharmaceutically acceptable carriers or diluents are demineralised or distilled water; saline solution; vegetable based oils such as peanut oil, safflower oil, olive oil, cottonseed oil, maize oil, sesame oil, arachis oil or coconut oil; silicone oils, including polysiloxanes, such as methyl polysiloxane, phenyl polysiloxane and methylphenyl polysolpoxane; volatile silicones; mineral oils such as liquid paraffin, soft paraffin or squalane; cellulose derivatives such as methyl cellulose, ethyl cellulose, carboxymethylcellulose, sodium carboxymethylcellulose or hydroxypropylmethylcellulose; lower alkanols, for example ethanol or iso-propanol; lower aralkanols; lower polyalkylene glycols or lower alkylene glycols, for example polyethylene glycol, polypropylene glycol, ethylene glycol, propylene glycol, 1,3-butylene glycol or glycerin; fatty acid esters such as isopropyl palmitate, isopropyl myristate or ethyl oleate; polyvinylpyrridone; agar; carrageenan; gum tragacanth or gum acacia, and petroleum jelly. Typically, the carrier or carriers will form from 10% to 99.9% by weight of the compositions.

[0100] Pharmaceutical forms suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The formulation must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The preventions of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin.

[0101] Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilisation. Generally, dispersions are prepared by incorporating the various sterilised active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze-drying technique which yield a powder of the active ingredient plus any additional desired ingredient from previously sterile-filtered solution thereof.

[0102] When the active agents are suitably protected they may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsule, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet. For oral therapeutic administration, the active compound may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 1% by weight of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 5 to about 80% of the weight of the unit. The amount of active compound in such therapeutically useful compositions is such that a suitable dosage will be obtained. Preferred compositions or preparations according to the present invention are prepared so that an oral dosage unit form contains between about 0.1 pg and 2000 mg of active.

[0103] Tablets, troches, pills, capsules and the like may also contain the components as listed hereafter: a binder such as gum, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, lactose or saccharin may be added or a flavouring agent such as peppermint, oil of Wintergreen, or cherry flavouring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellac, sugar or both. A syrup or elixir may contain sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavouring such as cherry or orange flavour. Of course, any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed. In addition, the oligonucleotides may be incorporated into sustained-release preparations and formulations.Therapeutic methods

[0104] Provided herein are methods for improving, increasing, restoring or retaining the integrity of the BBB in a subject (e.g. a subject with reduced or damaged BBB integrity and / or asubject with a disease or condition associated with BBB damage or loss of BBB integrity) ; for reducing or inhibiting the deposition or accumulation of amyloid-3 in the brain of a subject in need thereof; for increasing or promoting the clearance of amyloid-3 in the brain of a subject in need thereof; and / or for inhibiting or reducing senescence of cells (such as brain cells) in a subject in need thereof. The methods include the step of administering to a subject in need thereof an effective amount of an oligonucleotide described herein.

[0105] Typically, the subject has been diagnosed with a disease or condition associated with BBB damage or loss of BBB integrity, or has been determined to be likely to develop such as disease or condition. Diseases and conditions associated with BBB damage or loss of BBB integrity include neuroinflammatory and / or neurodegenerative diseases, including but are not limited to, dementia, multiple sclerosis (MS), motor neurone disease (MND) (e.g. amyotrophic lateral sclerosis (ALS), primary lateral sclerosis (PLS), progressive muscular atrophy (PMA), progressive bulbar palsy (PBP) and pseudobulbar palsy), Parkinson's disease and Huntington's disease. The dementia may be associated with or characterized by plaque deposition, or may have little or no associate with or evidence of amyloid-3 deposition. Non-limiting examples of dementia include Alzheimer's disease, Lewy body dementia (LBD), Parkinson's disease dementia (PDD), vascular dementia and frontotemporal dementia (FTD)). Other conditions associated with a loss of BBB integrity include traumatic brain injury (TBI), hypertension (including chronic hypertension) and elevated pulse pressure (e.g. at least 40 mm Hg, 45 mm Hg, 50 mm Hg, 55 mm Hg, 60 mm Hg, or 65 mm Hg), conditions that are also associated with dementia. Thus, in some examples, the subject has TBI and dementia, or hypertension and dementia, or elevated blood pressure and dementia.

[0106] Subjects with a neuroinflammatory and / or neurodegenerative disease or at risk for a neuroinflammatory and / or neurodegenerative disease include patients who have one or more biomarkers of disease severity, or which indicate a susceptibility to developing a neuroinflammatory and / or neurodegenerative disease. Such markers are known in the art and can be used by a clinician, optionally with other clinical signs of disease (e.g. mobility, cognition, etc.).

[0107] For example, where the disease is dementia including Alzheimer's disease, FTD, LBD, PDD and vascular dementia, the marker may be selected from one or more of a mutation in the gene encoding amyloid precursor protein (APP) and presenilins 1 and 2, mutations in the £4, 2 and 3 alleles of the apolipoprotein E (APOE) gene (APOE- s4, APOE- s2, APOE- s3), mutations in Triggering receptor expressed on myeloid cells 2 (TREM2) gene, MAPT gene, GRN gene, also called the PGRN gene, TARDBP gene, VCP gene and the CHMP2B gene. Elevated serum and / or CSF levels of a-synuclein, S100A9 and S100B, chromogranin, circulating DNA, heat shock proteins and amyloid may also be determined. In some examples, high blood pressure or hypertension is a risk factor for the development of dementia, particularly vascular dementia.

[0108] In another example, biomarkers for MND may be selected from one or more of e.g. SOD1, TDP-43, FUS, C9ORF72, ALS2, ALS4, ALS8, NEK1, UBQLN2, VCP, SETX, ANG, PFN1,MATR3, CHCHD10, TUBA4A, TBK1, C21orf2 and OPTN or an expression product thereof. In some embodiments for MND, the presence of cytoplasmic deposition of TDP-43-positive inclusions and / or elevated serum and / or CSF levels of neurofilaments, may also be determined.

[0109] In other embodiments, subjects at risk for a neuroinflammatory or neurodegenerative disease may also be identified by determining the presence of elevated levels of one or more of pro-inflammatory cytokines associated with disease, e.g. TNF, IL-l-a, IL-6, IFN-p, IL-ip, IL-8, IL-18, C-reactive protein (CRP), IL-17, chemokines, CD14+-high monocytes and inflammatory mediator mRNA transcripts in peripheral blood mononuclear cells (PBMC). In an embodiment, the disease is MND and the cytokine is selected from one or more of IL-6 or IL- 17. In an embodiment, the disease is Dementia and the cytokine is selected from IL-1, IL-6 and TNF-a.

[0110] In some examples, the subject has a disease associated with amyloid-p deposition, such as AD, CAA, LBD or TBI, or has been determined to be likely to develop AD, CAA or LBD. Thus, also provided are methods for treating a subject with AD, CAA, LBD or TBI, or treating a subject who is likely to develop AD, CAA or LBD, wherein the methods include the step of administering an effective amount of an oligonucleotide described herein to the subject. In some examples, the TBI is mild, moderate or severe TBI, or moderate-severe TBI.

[0111] In some examples, the subject has been diagnosed with AD, including early onset AD, late onset AD, familial AD, early stage AD and / or late stage AD. In other examples, the subject has been determined (such as by a clinician) to be likely to develop AD, e.g. to have a medium or high likelihood of developing AD, such as a 20%, 30%, 40%, 50% or more likelihood of developing AD. In other examples, the subject has been diagnosed with CAA (including probable CAA), or has been determined to be likely to develop CAA, e.g. to have a medium or high likelihood of developing CAA, such as a 20%, 30%, 40%, 50% or more likelihood of developing CAA.

[0112] Any suitable amount or dose of an oligonucleotide of the invention may be administered to a subject in need in accordance with the present invention. The therapeutically effective amount for any particular subject may depend upon a variety of factors including: the disease being treated and the severity of the disease; the activity of the oligonucleotide employed; the composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration; the route of administration; the rate of sequestration of the molecule or agent; the duration of the treatment; drugs used in combination or coincidental with the treatment, together with other related factors well known in medicine. One skilled in the art would be able, by routine experimentation, to determine an effective, non-toxic amount of protein conjugate to be employed.

[0113] The oligonucleotide can be administered alone or in conjunction with another therapy. The therapy may include another active agent, a device, a physical therapy, a cognitive therapy, an occupational therapy, or any other therapy that may retain or improve BBB integrity and / ortreat a disease or condition, including a neuroinflammatory disease, a neurodegenerative disease, hypertension and / or elevated pulse pressure.

[0114] In one example, the additional therapy includes the use of an active agent and / or device for reducing blood pressure and / or pulse pressure, including reducing blood pressure and / or pulse pressure in the cerebral vessels. Non-limiting examples of such agents include angiotensin-converting enzyme (ACE) inhibitors, Angiotensin II receptor blockers (ARBs) and calcium channel blockers. Non-limiting examples of such devices include those described in International patent publication number WO2021119737. These devices are placed around the external wall of a blood vessel (such as the carotid artery, e.g. the common carotid artery or the internal carotid artery), changing the geometry of the blood vessel so as to, for example, modify the dynamics of transmission of arterial blood pressure to the cerebral microvasculature and to absorb excess energy from the arterial pulse. In some examples, application of the active agent and / or device for reducing blood pressure and / or pulse pressure result in a reduction of blood pressure and / or pulse pressure to normal levels or ranges, e.g. systolic pressure of less than 120, 125, 130, 135 or 140 mm Hg, diastolic pressure of less than 80, 85 or 90 mm Hg, and / or pulse pressure of less than 40, 45, 50, 55 or 60 mm Hg.

[0115] In other examples, the additional therapy includes an active agent for treating dementia, and includes, for example, a cholinesterase inhibitors such as donepezil, rivastigmine or galantamine, memantine and lecanemab. In further example, the active agent is for treating MND, and includes, for example, riluzole, agents that block the interaction between CD40 and CD40 ligand, including antibodies that bind specifically to CD40 and / or CD40 ligand, and antiinflammatories. In further examples, the active agent is for treating Parkinson's disease, and includes levodopa, cardidopa-levodopa, dopamine agonists, monoamine oxidase B (MAO B) inhibitor, catechol O-methyltransferase (COMT) inhibitors, amantadine, and adenosine receptor antagonists. Additional active agents that can be administered for the treatment of MS include, for example, ofatumumab, teriflunomide, dimethyl fumarate, diroximel fumarate, monomethyl fumarate, fingolimod, siponimod, ozanimod and ponesimod.

[0116] The skilled addressee will recognise that in determining an appropriate and effective dosage range for administration to humans based on the mouse studies exemplified herein, dose escalation studies would be conducted. The skilled addressee would therefore appreciate that the above mentioned doses and dosage ranges are exemplary only based on the doses administered in the mouse studies exemplified herein, and the actual dose or dosage range to be employed in humans may be varied depending on the results of such dose escalation studies. Based on the data exemplified herein, the appropriate and effective dose or dosage range to be administered to humans can be determined by routine optimisation, without undue burden or experimentation.

[0117] In some examples, the subject is administered 1, 2, 3, 4, 5, 6, 7 or more doses of the oligonucleotide, over a period of days, weeks, months or years, e.g. every 1, 2, 3, 4, 6, 7, 8,9 10 or more days, weeks or months. In some examples, the subject is administered maintenance doses over a period of years.

[0118] Administration of an oligonucleotide of the invention, optionally with a further therapy, can improve, increase, restore or retain the integrity of the BBB, resulting in, for example, at least a 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100% or more improvement in the BBB integrity compared to prior to administration, and / or limiting any loss of BBB integrity to, for example, less than 30%, 25%, 20%, 15%, 10%, 5% or less over time. BBB integrity can be assessed by measuring BBB permeability using non-invasive imaging techniques such as magnetic resonance imaging, positron emission tomography and perfusion computed tomography (see e.g. Avsenik et al. Radiol Oncol. 2015 Jun; 49(2): 107-114 and Harris et al. Eur J Nucl Med Mol Imaging. 2023; 50(4): 1051-1083).

[0119] In some examples, administration of the oligonucleotide (and optional additional therapy) results in at least a 10%, 15%, 20%, 25%, 30%, 40%, 45% or 50% or more reduction in the amount of amyloid-p deposition in the brain compared to prior to administration. In other examples, administration of the oligonucleotide results in an inhibition or slowing of the rate of accumulation of amyloid-p deposition in the brain compared to prior to administration or compared to a subject with a comparable disease and levels of amyloid-p deposition but who has not been administered the oligonucleotide. In such instances, the rate or amount of amyloid-p deposition of may be reduced by at least 10%, 15%, 20%, 25%, 30%, 40%, 45% or 50% or more. The reduction in the amount or rate of amyloid-p deposition or accumulation in the brain may be in the vasculature of the brain, such as in the leptomeninges vessels or other small to medium sized vessels of the brain, and / or in the parenchyma of the brain. Moreover, the deposits of amyloid-p can comprise amyloid-p monomers and / or amyloid-p aggregates, such as amyloid- 8 oligomers, amyloid-p fibrils and / or amyloid-p plaques.

[0120] Levels of amyloid-p deposition in the brain can be assessed using any known method in the art. In some examples, the amyloid-p deposits are visualized and measured directly using brain imaging techniques such as positron emission tomography. In other examples, markers of brain amyloid-p deposition are used to indirectly measure the levels of amyloid-p deposition in the brain. These include, but are not limited to, the detection of amyloid-p (and in particular AP40 and / or A[J42) in cerebrospinal fluid (CSF) or plasma samples (see e.g. Li et al., Neurology. 2022 Feb 15; 98(7): e688-e699).

[0121] Administration of an oligonucleotide of the present disclosure may result in improvement or amelioration of any one or more symptoms of the disease or condition associated with loss of BBB integrity and / or with amyloid-p deposition. In particular examples, when the disease is a dementia (including dementia resulting from high blood pressure and / or pulse pressure), administration of an oligonucleotide of the present disclosure results in improvement of cognitive function or in a slowing in the rate of cognitive decline (e.g. of at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80% 90% or more). Cognitive assessment tools are well known and include, for example, the 10-point cognitive screener (10-CS), the 6-item cognitiveimpairment test (6CIT), the 6-item screener, the Memory Impairment Screen (MIS), the MiniCog, or Test Your Memory (TYM).Kits

[0122] Embodiments of the present invention also provide kits for use in accordance with the invention. For example, kits of the invention may contain one or more oligonucleotides disclosed herein, and optionally scrambled oligonucleotides for use as controls. Such kits may be used, for example, in medical or biological research activities, including investigations into amyloid-g deposition and plaque formation, accumulation or clearance. Kits according to the present invention may also include other components required to use the oligonucleotides, such as buffers and / or diluents. The kits typically include containers for housing the various components and instructions for using the kit components in the methods of the present invention.

[0123] In order that the invention may be readily understood and put into practical effect, particular preferred embodiments will now be described by way of the following non-limiting examples.

[0124] The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.ExamplesExample 1: General methodsOligonucleotides

[0125] Blockmirs were synthesized by RiboTask, Denmark. The sequences of these oligonucleotides are provided in Table 3. The oligonucleotides were formulated in PBS.Table 3. Oligonucleotide sequencesSingle underlining represents an LNA monomer; double underlining represents a 2' O-methyl RNA monomer; bold represents phosphorothiateMouse model of AD

[0126] The transgenic mouse model of AD used in the studies was APPswe / PSldE9 mice, one of the most extensively used transgenic mouse model of AD. These transgenic mice overexpress the human amyloid precursor protein gene (APP) with the Swedish mutation and the human presenilin-1 gene with deletion of exon 9, both familial AD genes, and are. Amyloid-g deposition can be seen in these mice as early as 4-6 months of age, with significant amyloid- P deposition by 9 months.

[0127] APP / PS1 transgenic mice and wild-type littermates were obtained from Jackson Labs USA and maintained by Australian BioResources Pty Ltd (ABR), NSW.Oligonucleotide delivery

[0128] APP / PS1 mice at 4 months (pre-plaque formation) and 6 or 7 months of age (postplaque formation) and wild-type controls were intravenously administered CD5-2 or control oligonucleotide at 30 mg / kg, or PBS alone. Mice were monitored for any adverse reaction for 20 minutes post-injection.Preparation and analysis of tissues

[0129] Mice were euthanased before cardiac perfusion was performed. In instances where biotin was used to detect and measure of vascular leak, the mice were injected with 100 mg / kg body weight of unconjugated biotin 1 hour prior to euthanasia.

[0130] The total period from euthanasia to cardiac perfusion was 10 minutes per mouse. Mice were perfused through the left ventricle compartment with 10 mL of cold IX PBS followed by 10 mL 1% PFA / 1X PBS. The brain cortices were collected and immersion-fixed with 4% PFA for 5 to 6 hours at 4 °C. In some studies, brains were cut sagittal through the midline into 2 halves. One half was fixed in cold 4% PFA in IX PBS for 4-5 hours, and the other half was fixed in methanol overnight in 4°C.

[0131] Tissue sections of 100 pm thickness were cut using a vibratome (Leica).

[0132] For PFA samples, the tissues were blocked overnight with 1% BSA in PBST (PBS with 1% Triton-X). Sections were stained overnight with antibodies for blood vessels (Isolectin-B4), endothelial cells (CD31), VE-cadherin, pericytes (PDGFRb, CD13), amyloid (D54D2), albumin, biotin leakage. After primary antibody incubation, the sections were washed extensively with trisbuffered saline 0.3% Triton-XlOO (TBST) and incubated with 1:500 Alexa Fluor 555, Alexa Fluor 594 and / or Alexa Fluor 647-preconjugated secondary antibodies (Molecular Probes, Invitrogen) for 2 hours at room temperature. Next, sections were washed three times with TBST and stained with DAPI for 10 minutes at room temperature. Finally, sections were washed once in PBS and cover-slipped with prolong Gold anti-fade mounting medium. Albumin detected at blood vessels located at top and bottom of the Z-stack was excluded as these leaks were considered an artefact of tissue cutting.

[0133] Methanol-fixed brain was sectioned at 100 m thick sagittal slices and stained with 1:200 rabbit anti-mouse Claudin-5 (Invitrogen #34-1600) and rat anti-mouse CD31 (BD #553370).

[0134] For quantitation of stained sections, tilescans of whole brain sections were imaged using confocal microscope at 20x magnification using Leica confocal microscope and Leica Application Suite (LAS) software. Expression of VE-cadherin and Claudin-5 expression in leptomeningeal vasculature were analysed using Image! software (version 2.14.0 / 1.54f). Regions of leptomeningeal layer was outlined and VE-cadherin or Claudin-5 expression in the blood vessels were thresholded and measured for mean intensity.VE-cadherin expression analysis

[0135] Human endothelial cells were cultured in medium 199 (Sigma-Aldrich, MO, USA) supplemented with 20% fetal bovine serum (FBS), 100 U / ml penicillin, 100 U / ml streptomycin, 15 pg / ml endothelial cell growth factor (BD Biosciences, MA, USA) and 15 pg / ml heparin (Sigma- Aldrich, MO, USA) at a humidified 37°C, 5%CO2 incubator. Oligonucleotides were transfected into cells using Hi perfect transfection reagent (Qiagen, Hilden, Germany), diluted in 37.5 pl OptiMEM (Thermo Fisher Scientific, MA, USA) and incubated overnight after which time medium was changed. The effect of oligonucleotides were assessed on cell lysates by Western blot analysis 48 hours after transfection.Example 2. Effect of CD5-2 on BBB integrity and amyloid-(3 deposition

[0136] Vascular leak was used as a marker or indication of BBB integrity in the APPswe / PSldE9 mouse model of AD. Vascular leak was first assessed by detecting the leak of biotin into the brain in 4 month old APPswe / PSldE9 mice that had been injected with biotin prior to euthanasia. The mice were administered the control blockmir, CD5-2 or PBS alone one week prior to euthanasia and preparation of brain tissue. As shown in Figure 1, APPswe / PSldE9 mice that were untreated exhibited vascular leak, indicating that the BBB and vasculature of these preplaque mice was impaired. In contrast, sections from APPswe / PSldE9 mice that received CD5-2 showed little or no biotin, indicating little or no vascular leak in these mice. No vascular leak was observed in wild-type mice, as expected (data not shown).

[0137] BBB and vascular integrity was also assessed using albumin and VE-cadherin as markers. APPswe / PSldE9 and wild-type mice (7 months old) were administered the control blockmir or CD5-2 one week prior to euthanasia and preparation of brain tissue sections. As shown in Figure 2, large albumin leaks were observed in control -treated APPswe / PSldE9 mice. By comparison, APPswe / PSldE9 mice that received CD5-2 showed reduced albumin leak.

[0138] In a further study, 6 month old APPswe / PSldE9 and wild-type mice were administered the control blockmir or CD5-2 one week prior to euthanasia and preparation of brain tissue sections. Sections were then stained to detect signs of amyloid-p deposition. As shown in Figure 3, significant amyloid-p deposition was detected on leptomeningeal vessels (LV) and in thebrain parenchyma in control-treated APPswe / PSldE9 mice, as expected. Surprisingly however, treatment of APPswe / PSldE9 mice with CD5-2 significantly reduced amyloid-3 deposition in the LV wall. When the brain tissues of these 6 month old mice was assessed for albumin leak and VE-cadherin expression, reduced VE-cadherin expression and significant albumin leak was observed in APPswe / PSldE9 mice that received the control blockmir, while APPswe / PSldE9 mice that received CD5-2 showed reduced albumin leak and restored VE-cadherin levels (similar to wild-type mice)(Figure 4).

[0139] This data clearly demonstrates that BBB and vascular integrity is impaired in APPswe / PSldE9 mice as early as 4 months, and is still present at 6-7 months along with amyloid- P deposition in the LV and brain parenchyma. Surprisingly, the administration of CD5-2 is sufficient to restore or improve BBB and vascular integrity in the brain. Moreover, administration of CD5-2 reduces amyloid-3 deposition. Thus, not only can CD5-2 reinstate the integrity of the BBB in APPswe / PSldE9 mice, it can also reduce the deposition or accumulation, and / or promote the clearance, of amyloid-3-Example 2. Effect of CD5-2, CD5-4 and CD5-9 on VE-cadherin expression.

[0140] CD5-2 has previously been shown to increase VE-cadherin expression and reduce vascular leak (see International Patent Publication No. W02014053014). The ability of two other related blockmirs, CD5-4 and CD5-9 (Figure 5C), to increase or promote VE-cadherin was tested and compared to CD5-2. CD5-4 and CD5-9 were designed for alternative and / or improved binding to the VE-cadherin 3'UTR (e.g. cross species binding), and / or to reduce or eliminate possible off- target binding (e.g. to ORC4 and / or FXR1). As shown in Figures 5A and B, both CD5-4 and CD5- 9 demonstrated comparable or even improved activity compared to CD5-2, significantly increasing VE-cadherin expression in endothelial cells, indicating that these blockmirs could be used in the same manner as has been shown for CD5-2, including to improve BBB and vascular integrity; and reduce deposition or accumulation of amyloid-3 and / or promote the clearance of amyloid-3- Indeed, further studies confirmed that CD5-4 has the expected activity (i.e. expected based on the activity of CD5-2) in vivo (data not shown).Example 3. Further assessment of effect of CD5-2 on vessels.

[0141] Further studies were performed to assess the effect of CD5-2 on the brain. APPswe / PSldE9 mice (6-7 months old) were administered CD5-2 or the control blockmir as described above, one week prior to euthanasia and preparation of brain tissue. Various analyses of the sections were performed to assess the effect of CD5-2 on expression of VE-cadherin and Claudin-5, albumin leak, leptomeningeal vessel density, amyloid deposition, and senescence.VE-cadherin and Claudin-5

[0142] Mice were perfused with cold PBS followed by 1% PFA as described above. Brains were cut sagittal through the midline into 2 halves. One half was drop-fixed in cold 4% PFA for4-5 hours, and the PFA fixed brain was then sectioned into 100 m thick sagittal slices and stained for VE-cadherin or albumin. The other half was dropped fixed in methanol overnight in 4°C, and the methanol-fixed brain was sectioned at 100 pm thick sagittal slices and stained for Claudin-5 and CD31. VE-cadherin or Claudin-5 expression in the leptomeningeal vasculature was measured as described above by confocal microscopy with tilescans of whole brain sections.

[0143] As shown in Figure 6, CD5-2 not only increased VE-cadherin in EC junctions, as expected, but also increased Claudin-5.Albumin leak

[0144] Brain slices were prepared as above and stained with goat anti-mouse Albumin and rat anti-mouse CD31. Tilescans of whole brain sections were imaged using confocal microscope at 20x magnification as described above, and Z-stacks of tilescans were compared between mice treated with the control blockmir (Scramble control) and CD5-2 for albumin leak. Albumin leak was defined as albumin leaking from whole vessels in the middle of the z-stacks. Leak at top or bottom of the brain slices or z-stacks were considered artifacts and were excluded. Once leak was established, the number of leaks in the cortex were counted per brain for each mouse per treatment group. This was then normalized to the vascular density of each brain.

[0145] As shown in Figure 7, administration of CD5-2 markedly reduced the number of albumin leaks.Leptomeningeal vessel density

[0146] Regions of interest was used to outline the leptomeningeal area in the brain using the Image! software. Using CD31 expression (as above; Figure 7), the area of CD31 positive vessels was measured to determine the leptomeningeal vessel area / density. As shown in Figure 8, CD5-2 markedly reduced the vessel density in APPswe / PSldE9 mice.Amyloid deposition

[0147] Brain slices were prepared as above and stained with rabbit anti-human beta amyloid. This antibody can recognise all species of human beta amyloid peptides including 1-42 and 1-40. Tilescans of whole brain sections were imaged using confocal microscope at 20x magnification using Leica confocal microscope and Leica Application Suite (LAS) software. Regions of the cortex were outlined using Image! software. Beta amyloid expression was thresholded in the cortex region and analysed for number of amyloid (cut-off of 5 pixels). The total number of amyloid deposited was measured relative to perimeter size of the cortex region. The leptomeningeal area was defined by the top layer on the cortex by DAPI nuclear stain. Amyloid deposition in this layer was also compared between treatment groups.

[0148] As shown in Figure 9, CD5-2 reduced amyloid deposition in both the leptomeninges and the parenchyma.Senescence

[0149] Senescent cells were detected by p21 and loss of laminBl. Brain slices prepared as above were stained for senescent markers p21 and LaminBl. Senescent cells were characterized by high p21 expression and loss of LaminBl nuclear envelope. DAPI was used to stain for cell nuclei and identification of p21 expression in the nuclei. Regions of leptomeningeal and penetrating vessels were imaged throughout the sagittal brain section at 40x magnification using Leica confocal microscope. The images were stacked and analysed using Image! software.

[0150] As shown in Figure 10, it was observed that CD5-2 reduced the number of senescent cells in APPswe / PSldE9 mice.

Claims

CLAIMS:

1. A method for reducing or inhibiting the deposition or accumulation of amyloid-g in the brain of a subject, comprising administering to the subject an effective amount of an oligonucleotide comprising a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG, to said RNA.

2. A method for increasing or promoting the clearance of amyloid-p in the brain of a subject, comprising administering to the subject an effective amount of an oligonucleotide comprising a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG, to said RNA.

3. The method of claim 1 or 2, wherein the subject has been diagnosed with Alzheimer's disease (AD) Cerebral amyloid angiopathy (CAA), Lewy body dementia (LBD), or traumatic brain injury (TBI), or has been determined to have a likelihood of developing AD, CAA or LBD.

4. A method of treating a subject with AD, CAA, LBD or TBI, or a subject likely to develop AD, CAA or LBD, comprising administering to the subject an effective amount of an oligonucleotide comprising a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG, to said RNA.

5. The method of any one of claims 1-4, wherein the oligonucleotide comprises a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 2, or SEQ ID NO: 2 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG, to said RNA.

6. The method of claim 5, wherein the miR-27a miRNA is hsa-miR-27a comprising the nucleotide sequence set forth in SEQ ID NO: 13.

7. The method of any one of claims 1 to 6, wherein the oligonucleotide comprises a contiguous sequence complementary to a sequence of at least or about 7 bases, at least or about 8 bases, at least or about 9 bases, at least or about 10 bases, at least or about 11 bases, at least or about 12 bases, at least or about 13 bases, at least or about 14 bases, at least or about 15 bases, at least or about 16 bases, at least or about 17 bases, at least or about 18 bases, at least or about 19 bases, at least or about 20 bases, at least or about 22 bases, at least or about 25 bases, at least or about 30 bases, or at least or about 35 bases of SEQ ID NO: 2, or SEQ ID NO: 2 comprising 1, 2 or 3 substitutions.

8. The method of any one of claims 1 to 7, wherein the oligonucleotide binds to positions 22-27 of SEQ ID NO: 2.

9. The method of any of claims 1 to 8, wherein base pairing between the oligonucleotide and SEQ ID NO: 2 includes positions 8-28, 8-27, 9-27, 10-27, 11-27, 12-27, 13-27, 14-27, 15-27, 16-27, 17-27, 18-27, 19-27, 20-27, 21-27, 9-28, 10-28, 11-28, 12-28, 13-28, 14-28, 15-28, 16- 28, 17-28, 18-28, 19-28, 20-28 or 21-28 of SEQ ID NO: 2.

10. The method of any one of claims 1 to 9, wherein the oligonucleotide comprises the sequence set forth in SEQ ID NO: 4.

11. The method of any one of claims 1 to 10, wherein the oligonucleotide comprises one or more modified nucleobases.

12. The method of claim 11, wherein the modified nucleobase is an LNA nucleobase, a UNA nucleobase or a 21O-methyl nucleobase.

13. The method of any one of claims 1 to 12, wherein the oligonucleotide comprises a sequence set forth in SEQ ID NO: 7.

14. The method of claim 4, wherein the treatment results in a reduction or inhibition of the deposition or accumulation of amyloid-p in the brain of the subject; and / or results in the promotion or increase in the clearance of amyloid-p in the brain of the subject.

15. The method of any one of claims 1, 2 or 14, wherein the reduction or inhibition of the deposition or accumulation of amyloid-p, and / or the promotion or increase in the clearance of amyloid-p, is in the vasculature of the brain and / or the paranchyma of the brain.

16. The method of claim 15, wherein the reduction or inhibition of the deposition or accumulation of amyloid-p, and / or the promotion or increase in the clearance of amyloid-p, is in the leptomeninges vessels of the brain.

17. The method of any one of claims 1, 2, and 14-16, wherein the amyloid-p is in the form of amyloid- 8 plaques.

18. The method of any one of claims 1 to 17, further comprising administering a further therapy.

19. The method of claim 18, wherein the further therapy comprises another active agent, a device, physical therapy, cognitive therapy, and / or occupational therapy.

20. The method of claim 18 or 19, wherein the further therapy comprises an active agent or device that reduces blood pressure and / or pulse pressure.

21. The method of any one of claims 1-20, wherein administration of the oligonucleotide and optionally the further therapy results in an improvement in cognition or a slowing in the rate of decline of cognition in the subject.

22. Use of an oligonucleotide in the preparation of a medicament for reducing or inhibiting the deposition or accumulation of amyloid-p plaques in the brain of a subject, wherein the oligonucleotide comprises a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNAcomprising a seed region comprising the sequence UCACAG, to said RNA.

23. Use of an oligonucleotide in the preparation of a medicament for increasing or promoting the clearance of amyloid-3 plaques in the brain of a subject, wherein the oligonucleotide comprises a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG, to said RNA.

24. The use of claim 22 or 23, wherein the subject has been diagnosed with Alzheimer's disease (AD), Cerebral amyloid angiopathy (CAA), Lewy body dementia (LBD) or traumatic brain injury (TBI), or has been determined to have a likelihood of developing AD, CAA or LBD.

25. Use of an oligonucleotide in the preparation of a medicament for treating a subject with Alzheimer's disease (AD), Cerebral amyloid angiopathy (CAA), Lewy body dementia (LBD) or traumatic brain injury (TBI), wherein the oligonucleotide comprises a contiguous sequence complementary to at least 8 contiguous bases of an RNA sequence comprising SEQ ID NO: 1, or SEQ ID NO: 1 comprising 1, 2 or 3 substitutions, wherein the oligonucleotide inhibits the binding of miR-27a, a variant thereof or a miRNA comprising a seed region comprising the sequence UCACAG, to said RNA.

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