Methods of treating cognitive impairment

Abstract

The present disclosure relates to the treatment of cognitive disorders, such as senescence-related cognitive impairment. In certain aspects, the disclosure describes methods of detecting one or more than one protein selected from the group consisting of DLL1, VNN2, VAV3, and SUMF1 from a sample obtained from a subject suffering from or suspected of suffering from a cognitive impairment. In certain embodiments, the cognitive impairment is caused by a degenerative disease of the nervous system, such as Alzheimer's disease. The method further includes identifying the subject as being likely or not likely to respond positively to plasma exchange therapy. In even other aspects, the present disclosure describes methods for treating cognitive impairment in a subject by plasma exchange therapy in which the subject is identified as likely or impossible to respond positively to the plasma exchange therapy based on particular protein expression data. The plasma exchange therapy may be full volume plasma exchange and / or low volume plasma exchange. Kits suitable for implementing the methods disclosed herein are also provided.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Serial No. 63 / 539,280, filed September 19, 2023, pursuant to 35 USC §119(e); the disclosure of which is incorporated herein by reference.

[0003] Reference to the electronic sequence list

[0004] The contents of the electronic sequence list (ALKA-034WO_SEQ_LIST.xml; size: 6935 bytes; creation date: September 5, 2024) are incorporated herein by reference in their entirety.

[0005] I. Introduction Technical Field

[0006] This invention relates to a method for treating cognitive impairment in patients. Background Technology

[0007] The following is provided for background information only and is not intended to be considered prior art of this invention.

[0008] Aging is a significant risk factor for a variety of human diseases, including cognitive impairment, cancer, arthritis, vision loss, osteoporosis, diabetes, cardiovascular disease, and stroke. Age-related neuronal degeneration is a major factor contributing to cognitive impairment. Therefore, aging remains the most important risk factor for dementia-related neurodegenerative diseases such as Alzheimer's disease (AD) (Bishop, N.A. et al., Neural mechanisms of ageing and cognitive decline. Nature 464(7288), 529-535 (2010); Heeden, T. et al., Insights into the ageingmind: a view from cognitive neuroscience. Nat. Rev. Neurosci. 5(2), 87-96 (2004); Mattson, MP. et al., Ageing and neuronal vulnerability. Nat. Rev. Neurosci. 7(4), 278-294 (2006)).

[0009] Aging affects all tissues and functions of the body, including the central nervous system, and neurodegeneration and functional decline such as cognitive or motor skills can severely impact quality of life. Treatments for cognitive decline, motor impairment, and neurodegenerative diseases have had limited success in preventing and reversing damage. Furthermore, while some treatments, such as plasma-based therapies or plasma exchange, have shown promise (Boada et al., Alzheimers Dement.; 16:1412-25, 2020), these treatments are not always successful. In other words, while some patients show positive outcomes after receiving such treatments for cognitive impairment, others do not. This results in a loss of valuable time and resources as the disease progresses.

[0010] Therefore, it is desirable to identify patients who may or may not respond positively to cognitive impairment treatment before implementing such treatments.

[0011] II. Summary of the Invention

[0012] In some aspects, the present invention provides methods for treating cognitive impairment. Such treatment may include plasma exchange therapy. In some cases, a subject is identified as potentially or potentially unlikely to respond positively to plasma exchange therapy. The determination of whether a subject is likely or unlikely to respond positively to plasma exchange therapy may be based on the detection of one or more proteins in a sample of the subject that indicate the subject's responsiveness to treatment. Some of these proteins may be one or more of the following: delta-like typical Notch ligand 1 (DLL1), vascular non-inflammatory molecule 2 (VNN2), guanine nucleotide exchange factor (VAV3), and sulfatase modifying factor 1 (SUMF1). In some embodiments, aptamer-based multiplex proteomics assays, such as SomaScan, are used. TM These proteins are detected using a detection method.

[0013] Therefore, in some aspects, the present invention provides a method for detecting a sample of one or more proteins selected from DLL1, VNN2, VAV3, and SUMF1; wherein the subject has or is suspected of having cognitive impairment.

[0014] Cognitive impairment can be caused by neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, and vascular dementia.

[0015] Other aspects of the invention provide kits comprising reagents for detecting one or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in a sample.

[0016] III. Merging by Reference

[0017] All publications and patent applications mentioned in this specification are incorporated herein by reference to the extent that each individual publication or patent application is specifically and individually indicated as incorporated by reference.

[0018] IV. Brief Description of the Attached Drawings

[0019] Figure 1 A schematic diagram illustrating an example of plasma exchange therapy is shown. Figure 1 In the described embodiments, the Alzheimer's Management By Albumin Replacement (AMBAR) medical research protocol includes a 6-week intensive period of routine therapeutic PE (TPE, also known as FPE – total plasma exchange), processing one plasma volume [≈2500 mL to 3000 mL], using 5% albumin for replacement (TPE once a week), followed by a 12-month maintenance period of low-volume plasma exchange (LVPE: removal of approximately 1 / 3 of plasma volume [≈690 mL to 880 mL]), using 20% ​​albumin or IVIG for replacement (LVPE once a month), applicable to all three active groups: one group uses 20 g of 20% Albutein for replacement in LVPE; while in the other two groups, Albutein 20% replacement (20 g or 40 g) and Flebogamma DIF 5% (10 g or 20 g) are used alternately. IVIG: Intravenous immunoglobulin; Albutein and FlebogammaDIF are commercially available products.

[0020] Figures 2A to 2B Proteomics analysis of the samples. Figure 2A This is a brief description of the statistical analysis for identifying protein biomarkers. Figure 2B It is a flowchart of the process of identifying one or more proteins that can indicate an object's responsiveness to treatment for cognitive impairment.

[0021] Figures 3A to 3C Examples of clinical changes predicted for stratified objects based on protein DLL1 are shown. Figure 3A This describes the relationship between baseline plasma DLL1 levels and improvement in the Clinical Dementia Rating-Sum of Boxes (CDR.Sb). Higher baseline DLL1 levels were associated with improvement in CDR.Sb (measured as change relative to baseline) (Spearman correlation coefficient (ρ) = -0.552, area under the curve (AUC) = 0.770 indicates better than baseline, AUC = 0.791 indicates no worse than baseline). Figure 3B The DLL1 receiver operating characteristics (ROC) are superior to the CDR.Sb baseline (bl) score in predicting study completion (EOS). Figure 3C This represents the estimated change in CDR.Sb in the stratified DLL1 group after plasma exchange therapy.

[0022] Figures 4A to 4C Examples of clinical changes predicted in hierarchical objects based on protein VNN2 are shown. Figure 4A This describes the relationship between baseline plasma VNN2 levels and improvement in CDR.Sb. Higher baseline VNN2 levels were associated with improvement in CDR.Sb (rho = -0.476, AUC = 0.761 indicates better than baseline, AUC = 0.726 indicates no worse than baseline). Figure 4B It is the ROC of VNN2, which outperforms the baseline score of CDR.Sb when used to predict EOS. Figure 4C This represents the estimated change in CDR.Sb in the stratified VNN2 group after plasma exchange therapy.

[0023] Figures 5A to 5C Some examples of clinical changes predicted in stratified subjects based on the protein VAV3 are shown. Figure 5A This describes the relationship between baseline plasma VAV3 levels and improvement in CDR.Sb. Higher baseline VAV3 levels were associated with improvement in CDR.Sb (rho = -0.494, AUC = 0.788 indicates better than baseline, AUC = 0.724 indicates no worse than baseline). Figure 5B It is the ROC of VAV3, which outperforms the bl score of CDR.Sb when used to predict EOS. Figure 5C This represents the estimated change in CDR.Sb in the stratified VAV3 group after plasma exchange therapy.

[0024] Figures 6A to 6C Some examples of clinical changes predicted in stratified subjects based on the protein SUMF1 are shown. Figure 6A This describes the relationship between baseline plasma SUMF1 levels and improvement in CDR.Sb. Higher baseline SUMF1 levels were associated with improvement in CDR.Sb (rho = -0.489, AUC = 0.696 indicates better than baseline, AUC = 0.756 indicates no worse than baseline). Figure 6B Figure 1 shows the ROC of SUMF1, used to predict EOS when the bl score is equal to or better than CDR.Sb. Figure C shows the estimated change in CDR.Sb in the stratified SUNF1 group after plasma exchange therapy.

[0025] V. Detailed Implementation

[0026] A. Introduction

[0027] This invention relates to the treatment of cognitive impairment, such as age-related cognitive impairment. In some aspects, the invention provides a method for detecting one or more proteins selected from the group consisting of DLL1, VNN2, VAV3, and SUMF1 in a sample obtained from a subject with or suspected of having cognitive impairment. In some embodiments, the cognitive impairment is caused by a neurodegenerative disease, such as AD. The method also includes identifying the subject as likely or unlikely to respond positively to plasma exchange therapy. In a further aspect, this disclosure describes a method for treating cognitive impairment in a subject by plasma exchange therapy, wherein the subject is identified as likely or unlikely to respond positively to said plasma exchange therapy based on specific protein expression data. Plasma exchange therapy can be full-volume plasma exchange and / or low-volume plasma exchange. In some cases, plasma exchange includes albumin exchange. Kits suitable for carrying out the methods of this disclosure are also provided.

[0028] Before describing the invention in detail, it should be understood that this disclosure is not limited to the specific methods or compositions described, as these can certainly vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, as the scope of this disclosure is limited only by the appended claims.

[0029] The publications discussed herein are provided only for their disclosure prior to the filing date of this application. Nothing herein should be construed as an admission that the invention is not entitled to precedence over such publication by virtue of a prior invention. Furthermore, the publication dates provided may differ from the actual publication dates, which require independent verification.

[0030] When a numerical range is provided, it should be understood that every intermediate value between the upper and lower limits of the range is included in this invention, unless the context explicitly indicates otherwise, each intermediate value should be as low as one-tenth of the lower limit unit. Every smaller range between any stated value or intermediate value within the range and any other stated value or intermediate value within the range is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range, and each range in which any, neither, or both are included is also included in this invention, but is subject to any specific exclusions from the range. When the range includes one or two limits, the range excluding any or both of those included limits is also included in this invention.

[0031] It should be noted that the claims may be drafted to exclude any optional elements. Therefore, this statement is intended as a premise for the use of exclusive terms such as "only" or "only" in relation to the elemental statement or "negative" limitation of the claims.

[0032] As will be apparent to those skilled in the art upon reading this disclosure, each individual embodiment described and illustrated herein has its own components and features, which can be readily separated from or combined with features of any other several embodiments without departing from the scope or spirit of the invention. Any of the described methods may be performed in the order of the events described or in any other logically possible order.

[0033] B. Definition

[0034] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of this invention, some possible and preferred methods and materials are described hereafter. All publications mentioned herein are incorporated herein by reference to disclose and describe methods and / or materials associated with the cited publications. It should be understood that, in the event of any conflict, this disclosure takes precedence over the disclosure of any incorporated publication.

[0035] It is important to note that, unless the context clearly specifies otherwise, as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural references. Thus, for example, “sample” can refer to multiple such samples.

[0036] In describing the methods of the present invention, the terms “host,” “object,” “individual,” and “patient” are used interchangeably and refer to any mammal requiring such treatment according to the disclosed methods. Such mammals include, for example, humans, sheep, cattle, horses, pigs, dogs, cats, non-human primates, mice, and rats. In some embodiments, the object is a non-human mammal. In some embodiments, the object is a farm animal. In other embodiments, the object is a pet. In some embodiments, the object is a mammal. In some cases, the object is a human. Other objects may include domestic pets (e.g., dogs and cats), livestock (e.g., cattle, pigs, goats, horses, etc.), rodents (e.g., mice, guinea pigs, and rats, for example, as animal models of disease), and non-human primates (e.g., chimpanzees and monkeys). Therefore, the object of the present invention includes, but is not limited to, mammals such as humans and other primates such as chimpanzees and other ape and monkey species; etc., wherein in some embodiments the object is a human. The term object is also intended to include any person or organism of any age, weight, or other physical characteristic, wherein the object can be an adult, child, infant, or newborn.

[0037] As used herein, “treatment” means reducing or eliminating cognitive impairment. Treatment can be therapeutically administered, i.e., administered after the onset of disease. Effects can include improving the cognitive performance of a subject with cognitive impairment. Therefore, as used herein, the term “treatment” encompasses any treatment of cognitive impairment in mammals and includes: (a) suppressing the disease, i.e., preventing its development; or (b) alleviating the disease, even if the disease subsides. Treatment can result in a variety of different physical manifestations, such as modulation of gene expression, tissue or organ rejuvenation, etc. Treatment can be administered during or after the onset of cognitive impairment. Treatment of ongoing cognitive impairment is of interest, wherein said treatment stabilizes or reduces adverse clinical symptoms in the patient. Such treatment can be administered before complete loss of function of the affected tissues. Treatment of said subjects can be administered during the symptomatic stage of the disease, and in some cases, after the symptomatic stage of the disease.

[0038] "Cognitive impairment" refers to an individual's cognitive abilities being impaired relative to healthy individuals (e.g., age-matched healthy individuals) or relative to an individual's abilities at an earlier point in time (e.g., 2 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, 5 years, or 10 years or more).

[0039] "Cognitive ability" or "cognition" refers to the mental processes that include attention and concentration, learning complex tasks and concepts, memory (acquiring, retaining, and retrieving new information in the short and / or long term), information processing (processing information collected by the five senses), visuospatial functions (visual perception, depth perception, using mental images, copying drawings, constructing objects or shapes), language production and comprehension, verbal fluency (word finding), problem-solving, decision-making, and executive functions (planning and prioritizing). "Cognitive decline" refers to the gradual decline of one or more of these abilities, such as a decrease in memory, language, thinking, or judgment.

[0040] In some implementations, cognitive impairment is defined as “age-related cognitive impairment.” As used herein, “age-related cognitive impairment” refers to cognitive impairment that is generally associated with aging, including, for example, cognitive impairment associated with the natural aging process, such as mild cognitive impairment (MCI); and cognitive impairment associated with age-related diseases, i.e., diseases that increase in frequency with aging, such as neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, vascular dementia, etc.

[0041] In some implementations, cognitive impairment in the subject is caused by neuroinflammation, for example, an increase in age-related neuroinflammation in the individual. "Neuroinflammation" refers to the biochemical and cellular responses of the nervous system to injury, infection, or neurodegenerative diseases. Such responses aim to mitigate triggering factors by mobilizing the central nervous system's immune system to defend against potential harm. Neurodegeneration occurs in the brain and spinal cord and is characterized by the loss of neuronal structure and function. Neuroinflammatory diseases or conditions or illnesses associated with neuroinflammation include, but are not limited to, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, etc.

[0042] C. Detection Method

[0043] Some aspects of the present invention provide a method for analyzing samples obtained from an object, the method comprising:

[0044] The sample was tested for one or more proteins selected from the following: DLL1, VNN2, VAV3, and SUMF1;

[0045] The subject in question suffers from or is suspected of suffering from cognitive impairment.

[0046] The DLL1 protein is present in various organisms, including humans. In humans, an instance of the DLL1 protein is described in the Uniprot database with ID number O00548. The sequence of the human DLL1 protein is as follows:

[0047]

[0048] Therefore, in some embodiments, the methods disclosed herein include detecting a protein having the sequence of SEQ ID NO: 1, or a protein having at least 70%, for example at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity with the sequence of SEQ ID NO: 1, from a sample of a subject suffering from or suspected of suffering from cognitive impairment. For example, those skilled in the art can readily identify homologs of the human DLL1 protein having the sequence of SEQ ID NO: 1 in an organism of interest, and then detect such DLL1 protein in a sample obtained from that organism. Such embodiments are within the scope of this disclosure.

[0049] The VNN2 protein is also present in various organisms, including humans. In humans, an instance of the VNN2 protein is described in the Uniprot database with ID number O95498. The sequence of the human VNN2 protein is as follows:

[0050]

[0051] Therefore, in some embodiments, the methods disclosed herein include detecting a protein having the sequence of SEQ ID NO: 2, or a protein having at least 70%, for example at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity with the sequence of SEQ ID NO: 2, from a sample of a subject with or suspected of having cognitive impairment. For example, those skilled in the art can readily identify homologs of the human VNN2 protein having the sequence of SEQ ID NO: 2 in an organism of interest, and then detect such VNN2 protein in a sample obtained from that organism. Such embodiments are within the scope of this disclosure.

[0052] Similarly, the VAV3 protein is present in a variety of organisms, including humans. In humans, an instance of the VAV3 protein is described in the Uniprot database with the ID Q9UKW4. The sequence of the human VAV3 protein is as follows:

[0053]

[0054] Therefore, in some embodiments, the methods disclosed herein include detecting a protein having the sequence of SEQ ID NO: 3, or a protein having at least 70%, for example at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity with the sequence of SEQ ID NO: 3, from a sample of a subject with or suspected of having cognitive impairment. For example, those skilled in the art can readily identify homologs of the human VAV3 protein having the sequence of SEQ ID NO: 3 in an organism of interest, and then detect such VAV3 protein in a sample obtained from that organism. Such embodiments are within the scope of this disclosure.

[0055] Similarly, the SUMF1 protein is present in various organisms, including humans. In humans, an instance of the SUMF1 protein is described in the Uniprot database with ID number P01042. The sequence of the human SUMF1 protein is as follows:

[0056]

[0057] Therefore, in some embodiments, the methods disclosed herein include detecting a protein having the sequence of SEQ ID NO: 4, or a protein having at least 70%, for example at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity with the sequence of SEQ ID NO: 4, from a sample of a subject with or suspected of having cognitive impairment. For example, those skilled in the art can readily identify homologs of the human SUMF1 protein having the sequence of SEQ ID NO: 4 in an organism of interest, and then detect such SUMF1 protein in a sample obtained from that organism. Such embodiments are within the scope of this disclosure.

[0058] The one or more proteins detected are also referred to as "target proteins" in this article.

[0059] In some cases, the methods disclosed herein include the qualitative detection of one or more target proteins in a sample.

[0060] Some non-limiting embodiments of qualitative detection of target proteins include determining the sequence of one or more target proteins and / or the post-translational modifications of one or more target proteins.

[0061] Such qualitative detection of one or more target proteins in a sample can be performed using any suitable method. The sample can be obtained from an individual with or suspected of having cognitive impairment. The sample can be any readily available biological sample. "Biological sample" refers both to a natural organism or a subset of its tissues and to homogenates, lysates, or extracts prepared from a subset of an organism or its tissues, including but not limited to, plasma, serum, cerebrospinal fluid, lymph, skin sections, respiratory sections, digestive tract sections, cardiovascular sections, and urogenital sections, tears, saliva, breast milk, blood cells, tumors, and organs. The biological sample can be any type of biological tissue, including healthy and diseased tissues (e.g., cancerous, malignant, necrotic, etc.). In some embodiments, the biological sample is a liquid sample, such as blood or its derivatives, such as plasma, tears, urine, semen, etc., wherein in some cases the sample is a blood sample, including whole blood, such as blood obtained by venipuncture or finger-prick sampling (wherein the blood may or may not be combined with any reagents, such as preservatives, anticoagulants, etc., before detection). Other embodiments of biological samples are well known to those skilled in the art, and such embodiments are within the scope of this invention. Methods for obtaining such samples from objects are also well known in the art, and such implementations fall within the scope of this disclosure.

[0062] In some cases, SomaScan is used. TM SomaScan analyzes the levels of the proteins described in this article in samples using a detection method. TMSome details of the detection method are described in reference Gold et al. (2010), PLoS One; 5(12): e15004, which is incorporated herein by reference in its entirety. In short, SomaScan... TM The assay uses aptamers called SOMAmers, which specifically bind to target protein classes. The assay involves binding SOMAmers to the target proteins in the sample and then isolating the SOMAmers, resulting in a complex mixture containing SOMAmers in proportion to the proteins present in the sample. The resulting mixture of SOMAmers is quantified using DNA microarray technology to produce a relative fluorescence unit (RFU) reading. Therefore, SomaScan... TM The detection method provides the levels of various proteins in a sample using "relative fluorescence units" (RFU) or derivatives of RFU, such as log2RFU.

[0063] Other non-limiting examples of methods for determining target protein sequences include mass spectrometry or Edman degradation using a protein sequencer.

[0064] Non-limiting examples of methods for determining post-translational modifications of target proteins include mass spectrometry and antibody-binding assays based on post-translational modifications, such as Western blotting or immunoassays.

[0065] In some cases, the methods disclosed herein involve the quantitative detection of one or more target proteins in a sample. Some non-limiting examples of the quantitative detection of one or more target proteins include immunoassays, mass spectrometry, and protein detection array analysis.

[0066] Immunoassays typically involve contacting a sample with a binding agent that specifically binds to a target protein, and then detecting the binding between the binding agent and the target protein. Such assays may include detecting a label conjugated to a binding agent or a label conjugated to a second binding agent that specifically binds to the binding agent and / or the binding agent-target protein complex. The binding agent is typically an antibody or an antigen-binding fragment of an antibody. The binding agent may also be an aptamer or a peptide-binding member.

[0067] Non-limiting embodiments of immunoassays include Western blot analysis, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), lateral flow immunoassay, particle-based immunoassay, quantum dot-based immunoassay, etc. Details of methods for performing some exemplary immunoassays are described in the review article Rizzo (2022), Chemosensors, 10(8), 326, the entire contents of which are incorporated herein by reference.

[0068] Mass spectrometry used to detect target proteins is typically designed to digest protein-containing samples into peptides, ionize these peptides, and analyze the ionized peptides to determine the identity of the digested protein. Due to the unique sequence of the protein, the specific digestion of the target protein produces unique digested and ionized peptides, which are identified in the mass spectrometer. The presence and quantity of these unique ionized peptides contribute to the identification of the target protein.

[0069] Non-limiting examples of mass spectrometry methods for detecting one or more target proteins include quadrupole mass spectrometry, time-of-flight mass spectrometry, magnetic sector mass spectrometry, electrostatic sector mass spectrometry, quadrupole ion trap mass spectrometry, and ion cyclotron resonance mass spectrometry. Details of some exemplary mass spectrometry-based detection methods are described in the review article Ma (2022), Molecules, 27, 6466, the entire contents of which are incorporated herein by reference.

[0070] Protein detection array analysis involves detecting the binding of a target protein in a sample to a binding agent (typically an antibody) located at a specific site on a support. The sample is brought into contact with such an array, and the binding of the target protein to the corresponding binding agent (e.g., an antibody located at a localized point on a solid support) is visualized, for example, using a detectable label. For instance, the captured target protein can be detected and / or quantified using a labeled secondary antibody, such as one labeled with a fluorescent dye.

[0071] As an alternative to binders located on a solid support, lysate microarrays involve immobilizing lysates of the sample onto a support, such as a nitrocellulose-coated glass slide. The immobilized proteins are then detected using a solution-bound binder capable of detecting label conjugations (e.g., a fluorescently labeled solution-phase-specific antibody). Differentially labeled binders, such as various fluorescently labeled antibodies targeting different target proteins, allow for multiplex detection of proteins, thus enabling the simultaneous detection of two or more proteins.

[0072] Non-limiting examples of protein detection array analysis include analytical protein microarrays, functional protein microarrays, and reversed-phase protein microarrays. Those skilled in the art can determine which protein microarrays are suitable for use with the methods disclosed herein. Details of methods for performing certain exemplary protein detection array analyses are described in the review article by Neagu et al. (2019), WorldAcad. Sci. J., 1:113-124, the entire contents of which are incorporated herein by reference.

[0073] In some cases, the method involves detecting one of DLL1, VNN2, VAV3, and SUMF1 in the sample.

[0074] In some cases, the method involves detecting any two of DLL1, VNN2, VAV3, and SUMF1 in the sample, such as the following combinations: DLL1 and VNN2; DLL1 and VAV3; DLL1 and SUMF1; VNN2 and VAV3; VNN2 and SUMF1; and VAV3 and SUMF1.

[0075] In other implementations, the method includes detecting any three of DLL1, VNN2, VAV3, and SUMF1 in a sample, such as the following combinations: DLL1, VNN2, and VAV3; DLL1, VAV3, and SUMF1; DLL1, VNN2, and SUMF1; and VNN2, VAV3, and SUMF1.

[0076] In some implementations, the method includes detecting all four types of molecules: DLL1, VNN2, VAV3, and SUMF1 in the sample.

[0077] In some cases, the method involves detecting only one protein among DLL1, VNN2, VAV3, and SUMF1 in the sample, without detecting other proteins except for certain control proteins in some cases.

[0078] In some cases, the method involves detecting only two proteins from DLL1, VNN2, VAV3, and SUMF1 in a sample, without detecting other proteins except for certain control proteins. For example, only the following protein combinations may be detected: DLL1 and VNN2; DLL1 and VAV3; DLL1 and SUMF1; VNN2 and VAV3; VNN2 and SUMF1; and VAV3 and SUMF1. In addition, one or more control proteins may be detected.

[0079] In other embodiments, the method includes detecting only three proteins of DLL1, VNN2, VAV3, and SUMF1 in a sample, without detecting other proteins except for certain control proteins. For example, the following protein combinations may be detected: DLL1, VNN2, and VAV3; DLL1, VAV3, and SUMF1; DLL1, VNN2, and SUMF1; and VNN2, VAV3, and SUMF1, and in addition, one or more control proteins may be detected.

[0080] In some implementations, the method includes detecting only four proteins in the sample: DLL1, VNN2, VAV3, and SUMF1, with the exception of certain control proteins in some cases, and no other proteins being detected.

[0081] As used herein, a "control protein" is a protein that cannot distinguish between subjects who may respond positively to plasma exchange therapy for the treatment of cognitive impairment and those who are unlikely to respond positively to plasma exchange therapy for the treatment of cognitive impairment. In other words, the level of the control protein is the same regardless of the subject's responsiveness to plasma exchange therapy for the treatment of cognitive impairment. Such proteins are known to those skilled in the art, or can be readily identified.

[0082] The methods disclosed herein for detecting one or more proteins may also include comparing the detection results with a threshold, a reference value, or a reference sample.

[0083] The threshold or reference value can be predetermined, for example, based on the concentration of one or more detected proteins in individuals known to respond to cognitive impairment treatment. For example, a retrospective testing of one or more proteins can be performed on a group of subjects who have responded to cognitive impairment treatment. These retrospective tests can be used to determine a threshold or reference value that truly indicates the likelihood of a subject responding to cognitive impairment treatment. In a specific implementation, the treatment is plasma exchange therapy, as described in detail below.

[0084] Alternatively, such a threshold or reference value can be predetermined, for example, based on the concentration of one or more detected proteins in individuals known to be unresponsive to cognitive impairment treatment. For instance, a retrospective testing of one or more proteins can be performed on a group of subjects unresponsive to cognitive impairment treatment. These retrospective tests can be used to determine a threshold or reference value that definitively indicates the likelihood of a subject responding to cognitive impairment treatment. In a specific implementation, the treatment is plasma exchange therapy, which is described in detail below.

[0085] Reference samples may contain one or more target proteins at known concentrations to indicate the subject’s responsiveness to plasma exchange therapy for the treatment of cognitive impairment.

[0086] The samples used to detect one or more target proteins in the methods disclosed herein can be obtained from subjects with cognitive impairment. Alternatively, the samples used to detect one or more target proteins in the methods disclosed herein can be obtained from subjects suspected of having cognitive impairment.

[0087] As used in this article, “subjects with cognitive impairment” refers to subjects who exhibit symptoms of cognitive impairment.

[0088] As used herein, “an individual suspected of having cognitive impairment” refers to a subject exhibiting symptoms of other conditions associated with cognitive impairment. For example, as discussed in detail below, a subject may exhibit some signs of a neurodegenerative disease, such as Alzheimer's disease or Parkinson's disease, but has not yet developed cognitive impairment. Such subjects are included in the scope of the term “subject suspected of having cognitive impairment.”

[0089] In some implementations, cognitive impairment is caused by neurodegenerative diseases. Non-limiting examples of neurodegenerative diseases that may cause cognitive impairment include Alzheimer's disease (AD), Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis, glaucoma, myotonic dystrophy, and vascular dementia. These diseases will be described below, and further information about them is well known in the art.

[0090] In the specific implementation plan, the neurodegenerative disease is AD, that is, the subject suffers from cognitive impairment caused by AD.

[0091] In some implementations, the samples are obtained from subjects who are candidates for plasma exchange therapy to treat cognitive impairment.

[0092] Any suitable sample from the subject can be tested according to the methods disclosed herein. In a specific implementation, the sample is a blood sample, serum sample, plasma sample, or cerebrospinal fluid sample.

[0093] In some cases, the sample may be aqueous humor, hyaline fluid, bile, chyle, endolymph, perilymph, lymph, mucus (including nasal secretions and phlegm), pericardial fluid, peritoneal fluid, pleural fluid, pus, rheum, saliva, sputum, or synovial fluid. Other examples of biological samples are well known to those skilled in the art, and such embodiments are within the scope of this invention. Methods for obtaining such samples from an object are also well known in the art, and such embodiments are within the scope of this disclosure.

[0094] D. Diagnostic methods

[0095] In addition to detecting one or more proteins in samples obtained from subjects with or suspected of having cognitive impairment, certain aspects of the invention provide for determining, based on the detection results, whether a subject is likely or unlikely to respond positively to a therapy intended to treat the subject's cognitive impairment. In some embodiments, the treatment is plasma exchange therapy. Details of plasma exchange therapies applicable according to the invention will be discussed in detail below and are applicable to the diagnostic methods described herein.

[0096] In some implementations, subjects are identified as potentially or potentially unlikely to respond positively to plasma exchange therapy for the treatment of cognitive impairment based on the detection results of one or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in the subject sample. For example, the detection results of DLL1 in the subject sample may identify subjects as potentially or potentially unlikely to respond positively to plasma exchange therapy for the treatment of cognitive impairment. Similarly, the detection results of VNN2 in the subject sample may identify subjects as potentially or potentially unlikely to respond positively to plasma exchange therapy for the treatment of cognitive impairment. Furthermore, the detection results of VAV3 in the subject sample may identify subjects as potentially or potentially unlikely to respond positively to plasma exchange therapy for the treatment of cognitive impairment. Moreover, the detection results of SUMF1 in the subject sample may identify subjects as potentially or potentially unlikely to respond positively to plasma exchange therapy for the treatment of cognitive impairment.

[0097] In other embodiments, based on the detection results of two or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in a subject sample, a subject can be identified as potentially or unpromisably responsive to plasma exchange therapy for the treatment of cognitive impairment. Such combinations include: DLL1 and VNN2; DLL1 and VAV3; DLL1 and SUMF1; VNN2 and VAV3; VNN2 and SUMF1; and VAV3 and SUMF1.

[0098] Additionally, based on the detection results of three or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in the subject sample, the subject can be identified as potentially or unlikely to respond positively to plasma exchange therapy used to treat cognitive impairment. Such combinations include: DLL1, VNN2, and VAV3; DLL1, VAV3, and SUMF1; DLL1, VNN2, and SUMF1; and VNN2, VAV3, and SUMF1.

[0099] Furthermore, based on the detection results of all proteins DLL1, VNN2, VAV3, and SUMF1 in the subject samples, the subjects can be identified as potentially or unpromising to respond positively to plasma exchange therapy used to treat cognitive impairment.

[0100] In some implementations, determining whether a subject is likely or unlikely to respond positively to a therapy for treating cognitive impairment based on test results includes comparing the test results with thresholds or reference values.

[0101] As described above, such thresholds or reference values ​​can be predetermined, for example, based on the concentration of one or more detected proteins in individuals known to respond to cognitive impairment treatment. For instance, retrospective testing of one or more proteins can be performed on a group of subjects who have responded to cognitive impairment treatment. These retrospective tests can be used to determine thresholds or reference values ​​that definitively indicate the likelihood of a subject responding to cognitive impairment treatment. In a specific embodiment, the therapy is plasma exchange therapy, which is described in detail below.

[0102] As described above, such thresholds or reference values ​​can be predetermined, for example, based on the concentration of one or more detectable proteins in individuals known to be unresponsive to cognitive impairment treatment. For instance, retrospective testing of one or more proteins can be performed on a group of subjects who have not responded to cognitive impairment treatment. These retrospective tests can be used to determine thresholds or reference values ​​that definitively indicate the likelihood of a subject responding to cognitive impairment treatment. In a specific implementation, the therapy is plasma exchange therapy, which is described in detail below.

[0103] Reference samples may contain one or more target proteins at known concentrations to indicate the subject’s responsiveness to plasma exchange therapy for the treatment of cognitive impairment.

[0104] In some implementations, an object is identified as potentially or unpromising to respond positively to plasma exchange therapy for the treatment of cognitive impairment, based on the level of increase of one or more proteins detected in a sample obtained from the object relative to a threshold, reference value, or reference sample.

[0105] As mentioned above, in some cases, SomaScan is used. TM The detection method analyzed the protein levels described in this article in patient samples. Tables 2 and 3 below provide the results obtained using SomaScan. TM Certain thresholds of proteins quantified by detection methods indicate a subject's responsiveness to cognitive impairment treatments.

[0106] Therefore, through SomaScan TM A log2RFU value of DLL protein in a sample from the subject, determined by a detection method, of at least 9 to 13, such as at least 9, at least 10, at least 11, at least 12, at least 13, or at least 11.2, indicates that the subject may be in a positive response to a therapy used to treat cognitive impairment, such as plasma exchange therapy. In some cases, plasma exchange therapy, such as... Figure 1 Or as described in the experimental section below.

[0107] Similarly, via SomaScan TMA detection method determining a log2RFU value of VNN2 protein in a sample from the subject of at least 8 to 12, such as at least 8, at least 9, at least 10, at least 11, at least 12, or at least 10.6, indicates that the subject may be in a positive response to a therapy used to treat cognitive impairment, such as plasma exchange therapy. In some cases, plasma exchange therapy, such as... Figure 1 Or as described in the experimental section below.

[0108] In addition, through SomaScan TM A log2RFU value of VAV3 protein in a sample from the subject, determined by a detection method, of at least 8 to 12, such as at least 8, at least 9, at least 10, at least 11, at least 12, or at least 10.4, indicates that the subject may be in a positive response to a therapy used to treat cognitive impairment, such as plasma exchange therapy. In some cases, plasma exchange therapy, such as... Figure 1 Or as described in the experimental section below.

[0109] Furthermore, through SomaScan TM A log2RFU value of SUMF1 protein in a sample from the subject, determined by a detection method, of at least 10 to 14, such as at least 10, at least 11, at least 12, at least 13, at least 14, or at least 12.1, indicates that the subject may be in a positive response to a therapy used to treat cognitive impairment, such as plasma exchange therapy. In some cases, plasma exchange therapy, such as... Figure 1 Or as described in the experimental section below.

[0110] As mentioned above, the protein levels described in this article can also be measured using methods other than SomaScan. TM Determined using techniques other than detection methods. Based on SomaScan. TM By analyzing the detection results and the associated log2RFU value of the protein described herein, a skilled technician can determine the actual protein concentration that indicates whether a patient is likely or unlikely to respond positively to a therapy used to treat cognitive impairment, such as plasma exchange. Certain such techniques have been described above in the discussion of suitable methods for the qualitative detection of one or more target proteins in a sample, and such embodiments fall within the scope of this disclosure.

[0111] When using certain types of SomaScan TMWhen using methods other than detection methods, those skilled in the art can determine appropriate thresholds or reference values ​​or reference samples that can be used to compare detection results to determine an increase in the level of one or more proteins. For example, such a threshold or reference value might indicate the highest level of one or more proteins in cognitive impairment, which might indicate responsiveness or non-responsiveness to plasma exchange therapy for cognitive impairment. Therefore, a level of one or more proteins above this threshold or reference value indicates that the subject has responded to plasma therapy for the treatment of cognitive impairment.

[0112] In some cases, the detection result of a protein can be compared with a threshold or reference value for the corresponding protein. As those skilled in the art will recognize, the control sample or reference value used for comparison will depend on the sample type, such as a blood sample or a cerebrospinal fluid sample.

[0113] In some other cases, the detection results of two, three, or four proteins can be compared with the corresponding threshold or reference value for combinations of two, three, or four proteins. When comparing the results of two or more proteins, certain statistical analyses can be performed on the results of individual proteins to obtain the combined results of two or more proteins, which are then compared with the threshold or reference value.

[0114] For example, a combination of two proteins may show a statistically significant increase in level when considered as a combination, while one of the proteins may show no change or even a decrease in level. Similarly, a combination of three proteins may show a statistically significant increase in level when considered as a combination, while one or two of the proteins may show no change or even a decrease in level. Likewise, a combination of four proteins may show a statistically significant increase in level when considered as a combination, while one, two, or three of the proteins may show no change or even a decrease in level. Statistical analyses used to generate combined values ​​for two or more protein levels are well known in the art. For example, in some cases, protein ratios, as well as multivariate linear and nonlinear models, can be used to compare protein expression data.

[0115] Statistical analysis

[0116] The following describes some of the terminology used in the statistical analysis of data obtained from bodily fluid samples used to analyze subjects. Based on the subjects' biomarker levels, these statistical analyses can be used to identify whether subjects are likely or unlikely to respond positively to treatment for cognitive impairment.

[0117] First, treatment benefit was confirmed by comparing changes in clinical outcomes between the treatment and placebo groups. Then, candidate proteomic biomarkers were identified by significant Spearman correlation coefficients between baseline protein levels measured by certain assays (here, the SomaScan assay, denoted as log2(RFU)) and altered cognitive outcomes at the end of the study. These candidate proteomic biomarkers were assessed by receiver operating characteristic (ROC) and measured by the area under the ROC curve (AUC) to indicate whether subjects would have better cognitive outcomes (with treatment benefit) or not (no treatment benefit) after treatment. ROC is a graph of sensitivity versus specificity for multiple thresholds of a biomarker. ROC is used to show the predictive power of a continuous biomarker for binary outcomes, such as treatment benefit or no treatment benefit.

[0118] AUC represents the probability of correctly ranking patients with or without treatment benefit based on continuous biomarkers. Generally, an AUC > 70% is considered acceptable, and an AUC > 80% is considered to have excellent predictive power.

[0119] Then, based on the optimal decision threshold (here, the optimal cutoff point or reference value for baseline protein levels) determined by the Youden's index, which maximizes accuracy (overall correct classification rate, as shown in Table 1 below), high biomarker levels (here, baseline protein levels) and low biomarker levels are defined. Finally, the optimal binary biomarker is determined based on its predictive power as measured by accuracy, specificity, and sensitivity (the calculation methods of which are explained below and shown in Table 1).

[0120] The terms A1, B1, A0, and B0 used in the following paragraphs are shown in Table 1 below:

[0121]

[0122] Accuracy represents the percentage of correctly classified subjects: (number of subjects who truly benefited from treatment + number of subjects who did not truly benefit from treatment) / total number of predictions. ((A1 + B0) / N, N = total number of predictions).

[0123] Specificity represents the true negative rate, which is the percentage of subjects who actually do not benefit from treatment out of the total number of subjects predicted not to benefit from treatment (B0 / (B1 + B0)).

[0124] Sensitivity represents the true positive rate, which is the percentage of subjects who actually have a treatment benefit out of the total number of subjects predicted to have a treatment benefit (A1 / (A1 + A0)).

[0125] These identified biomarkers will then be validated through computer simulations, such as testing in other clinical outcomes, testing in randomly reselected samples, testing at other visits, and / or testing in similar clinical trials.

[0126] According to the methods described herein, at least 60%, for example at least 70%, at least 80%, or at least 90% of the subjects identified as potentially likely to respond positively to treatment for cognitive impairment actually responded positively to the treatment. In some cases, plasma exchange therapy, such as Figure 1 Or as described in the experimental section below.

[0127] Conversely, according to the methods described herein, at least 60%, for example at least 70%, at least 80%, or at least 90% of patients identified as unlikely to respond positively to treatment for cognitive impairment actually do not respond positively to treatment. In some cases, plasma exchange therapy, such as Figure 1 Or as described in the experimental section below.

[0128] In some cases, treatments such as plasma exchange therapy are not administered to individuals who are deemed unlikely to respond positively to treatments for cognitive impairment, such as plasma exchange therapy.

[0129] In some cases, the baseline biomarker levels that indicate whether subjects are likely to respond positively to plasma therapy for the treatment of cognitive impairment are shown in Table 2 below:

[0130] Table 2. Baseline biomarker levels in blood indicating whether subjects are likely to have a positive (better than baseline) response to plasma therapy for the treatment of cognitive impairment (such as the plasma therapy described in the Experiments section below).

[0131]

[0132] Table 3. Baseline biomarker levels in blood indicating whether the subject is likely to respond positively to plasma therapy for the treatment of cognitive impairment (e.g., plasma therapy described in the Experiment section below) (deterioration less than 1 point – a clinically significant change in the total CDR box score).

[0133]

[0134] In some embodiments, the method may include providing a report indicating whether the subject is likely to respond to plasma therapy for the treatment of cognitive impairment. In some embodiments, this step may involve calculating a score based on the detection results of one or more proteins, the score being correlated with responsiveness, and may be numerical, such as a probability, likelihood, or a score out of 10. In these embodiments, the method may include inputting the amount of each protein into one or more algorithms or calculations, executing said algorithms or calculations, and receiving a score based on the calculations. In these embodiments, other measurements of the subject, such as whether the subject is male or female, the subject's age, the degree of current cognitive impairment, etc., may be considered and incorporated into the algorithm or calculation.

[0135] In some implementations, the method may involve creating a report, for example electronically, and forwarding it to physicians or other healthcare professionals to help determine appropriate course of action, such as identifying individuals suitable for receiving plasma exchange therapy to treat cognitive impairment. This report may be used in conjunction with other indicators to determine appropriate course of action.

[0136] In some cases, a report may be forwarded to a “remote location,” which refers to a location other than the location where the report was generated. For example, a remote location could be another location within the same city (e.g., an office, laboratory, etc.), another location in a different city, another location in a different state, another location in a different country, etc. Therefore, when one item is indicated as being “remote” to another item, the two items may be in the same room but separated, or at least in two different rooms or different buildings, and may be at least one mile, ten miles, or at least one hundred miles apart. “Communication” information refers to data representing that information transmitted in the form of electrical signals through a suitable communication channel (e.g., a private or public network). “Forwarding” an item refers to any means of moving the item from one location to another, whether by physical transport of the item or other means (where possible), and at least in terms of data, includes physically transporting the medium carrying the data or communicating the data. Examples of communication media include radio or infrared transmission channels, and network connections to another computer or networked device, the Internet, or information including email transmissions and records on websites, etc. In some implementations, the report may be analyzed by an MD or other qualified medical professional, and the report based on the image analysis results may be forwarded to the recipient who provided the sample.

[0137] In computer-related implementations, the system may include a computer that includes a processor, storage components (i.e., memory), a display component, and other components typically found in general-purpose computers. The storage components store information accessible to the processor, including instructions executable by the processor and data that can be retrieved, manipulated, or stored by the processor.

[0138] The storage component includes instructions for determining, using the detection results of one or more proteins as described above, whether a subject is likely to respond to plasma exchange therapy for the treatment of cognitive impairment. A computer processor is connected to the storage component and configured to execute the instructions stored in the storage component to receive patient data and analyze the patient data according to one or more algorithms or calculations. A display component can display information about the patient's responsiveness to plasma therapy for the treatment of cognitive impairment.

[0139] Storage components can be any type capable of storing information accessible to the processor, such as hard drives, memory cards, ROM, RAM, DVDs, CD-ROMs, USB flash drives, writable memory, and read-only memory. The processor can be any well-known processor, such as those from Intel. Alternatively, the processor can be a dedicated controller, such as an ASIC.

[0140] Instructions can be any set of instructions that are executed directly by the processor (e.g., machine code) or indirectly (e.g., a script). In this regard, the terms "instruction," "step," and "program" are used interchangeably herein. Instructions can be stored as object code for direct processor processing or in any other computer language, including scripts or sets of independent source code modules that are interpreted on demand or pre-compiled.

[0141] Data can be retrieved, stored, or modified by a processor according to instructions. For example, although the system disclosed herein is not limited to any particular data structure, data can be stored in a computer register, in a relational database as a table with multiple distinct fields and records, an XML document, or a flat file. Data can also be formatted in any computer-readable format, such as, but not limited to, binary values, ASCII, or Unicode. Furthermore, data can contain any information sufficient to identify the relevant information, such as numbers, descriptive text, proprietary code, pointers, references to data stored in other memory (including other network locations), or information about functions used to calculate the relevant data.

[0142] E. Treatment methods

[0143] In addition to determining whether a subject is likely to respond positively to a therapy for cognitive impairment, certain aspects of the invention also provide the ability to treat a subject's cognitive impairment by administering the therapy to the subject.

[0144] Therefore, certain aspects of this disclosure provide a method for treating cognitive impairment in a subject, the method comprising administering plasma exchange therapy to the subject, wherein the subject is identified as potentially or potentially unable to respond positively to plasma exchange therapy for treating the cognitive impairment.

[0145] In some cases, based on the detection results of one or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in the subject sample, the subject may or may not respond positively to plasma exchange therapy used to treat the cognitive impairment.

[0146] The details of identifying whether an object is likely or unlikely to respond positively to plasma exchange therapy for the treatment of cognitive impairment, based on the detection results of one or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in the object sample, are described in the "Diagnostic Methods" section above.

[0147] In short, based on the detection results of one, two, three, or four proteins from DLL1, VNN2, VAV3, and SUMF1 in the subject sample, the subject can be identified as potentially or unpromising to respond positively to plasma exchange therapy for the treatment of cognitive impairment.

[0148] Based on the increase in the levels of one, two, three, or four proteins detected in a sample obtained from the subject relative to a reference value or reference sample, an object can be identified as potentially or unpromising to respond positively to plasma exchange therapy for the treatment of cognitive impairment.

[0149] "Plasma exchange therapy" refers to replacing all or part of a patient's plasma with a plasma exchange solution. Typically, during plasma exchange therapy, blood is gradually removed from the patient's body, blood components (such as blood cells and platelets) are separated from the plasma, the plasma exchange solution is mixed with the separated blood components, and the resulting mixture is then returned to the patient.

[0150] Plasma exchange therapy may include replacing substantially all of a subject's plasma with a plasma exchange solution, referred to herein as "total plasma exchange". In total plasma exchange, substantially all of a subject's plasma, for example, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% of the plasma, is replaced by the plasma exchange solution. In some cases of total plasma exchange, the plasma exchange solution is an albumin solution containing 3% w / v to 10% w / v albumin, such as 3% w / v, 4% w / v, 5% w / v, 6% w / v, 7% w / v, 8% w / v, 9% w / v, or 10% w / v albumin solutions. In one embodiment, the plasma exchange solution used for total plasma exchange is a 5% w / v albumin solution.

[0151] Plasma exchange therapy may also involve replacing a portion of the subject's plasma with a plasma exchange solution, referred to herein as "low-volume plasma exchange." In low-volume plasma exchange, 10% w / v to 50% w / v, such as 15% w / v to 45% w / v, 20% w / v to 40% w / v, 25% w / v to 35% w / v, or about 30% w / v, of the subject's plasma is replaced by a plasma exchange solution. In some cases of low-volume plasma exchange, the plasma exchange solution is an albumin solution containing 15% w / v to 25% w / v albumin, such as 15% w / v, 16% w / v, 17% w / v, 18% w / v, 19% w / v, 20% w / v, 21% w / v, 22% w / v, 23% w / v, 24% w / v, or 25% w / v albumin.

[0152] In some implementations, plasma exchange therapy includes a course of total plasma exchange. In some cases, a course of total plasma exchange includes weekly sessions for 1 to 8 weeks, such as weekly sessions for 1, 2, 3, 4, 5, 6, 7, or 8 weeks. In some such implementations, the plasma exchange solution contains 5% w / v albumin.

[0153] In other embodiments, plasma exchange therapy includes a course of low-volume plasma exchange using a second albumin solution following a course of total plasma exchange. The course of low-volume plasma exchange may be administered monthly for at least one month to at least ten months, for example, monthly for 12 to 16 months, such as 12, 13, 14, 15, or 16 months. In some such embodiments, the plasma exchange solution contains 20% w / v albumin.

[0154] In specific implementation plans, plasma therapy includes Figure 1The treatments shown are as follows. Such treatments include weekly total plasma exchange for 6 weeks, with the plasma exchange solution containing 5% w / v albumin. Following this total plasma exchange course, the plasma exchange therapy includes monthly low-volume plasma exchange therapy for 10 to 16 months, such as 10, 11, 12, 13, 14, 15, or 16 months. The low-volume plasma exchange therapy involves replacing 15% to 25%, such as 20%, of the recipient's plasma with a plasma exchange solution containing 20 to 40 grams of albumin (e.g., 20, 30, or 40 grams of albumin). In some cases, the low-volume plasma exchange therapy involves replacing 15% to 25%, such as 20%, of the recipient's plasma with a plasma exchange solution containing 20 to 40 grams of albumin (e.g., 20, 30, or 40 grams of albumin) and 10 to 20 grams of immunoglobulin (e.g., 10, 15, or 20 grams of immunoglobulin).

[0155] In some cases, plasma exchange therapy includes weekly total plasma exchange for 6 weeks, with the plasma exchange solution containing 5% w / v albumin. For example, during total plasma exchange, 2500 to 3000 ml of patient plasma is replaced with an equal volume of 5% albumin solution (e.g., 5% albutein). Following total plasma exchange, plasma exchange therapy includes monthly low-volume plasma exchange for 10 to 16 months, e.g., 10, 11, 12, 13, 14, 15, or 16 months. During low-volume plasma exchange, 600 to 900 ml (e.g., 650 to 880 ml) of patient plasma is replaced with 100 or 200 ml of 20% albumin (e.g., 20% albutein). In some cases, low-volume plasma exchange therapy includes replacement with a 20% albumin solution (e.g., Albutein 20%) containing 20 to 40 grams (e.g., 20, 30, and 40 grams) of albumin. In some cases, albumin replacement is performed alternately with immunoglobulin replacement, for example, with 10 to 30 grams, such as 10, 20 or 30 grams of 5% immunoglobulin solution (e.g., Flebogamma DIF 5%).

[0156] Plasma replacement solution

[0157] In some cases, plasma exchange therapy may be performed using any of the plasma exchange solutions described below.

[0158] Albumin plasma products (APP) fall into two categories: plasma protein fraction (PPF) and human albumin solution (HAS). PPF is derived from processes with higher yields than HAS, but the minimum albumin purity of PPF is lower than that of HAS (PPF > 83%, HAS > 95%) (Production of human albumin solution: a continually developing colloid, P. Matejtschuk et al., British J. of Anaesthesia 85(6): 887-95, at 888 (2000)). In some cases, the albumin purity of PPF is between 83% and 95%, or between 83% and 96%. Albumin purity can be determined by electrophoresis or other quantitative methods, such as mass spectrometry.

[0159] Those skilled in the art will recognize that several commercial sources of PPF (“commercial PPF formulations”) have existed or exist now. These include Plasma-Plex™ PPF (Armour Pharmaceutical Co., Tarrytown, NY), Plasmanate™ PPF (Grifols, Clayton, NC), Plasmatein™ (Alpha Therapeutics, Los Angeles, CA), and Protenate™ PPF (Baxter Labs, Inc., Deerfield, IL).

[0160] Those skilled in the art will also recognize that several commercial sources of HAS (“commercial HAS formulations”) have existed or exist now. These include Albaminar™ (CSL Behring), AlbuRx™ (CSL Behring), Albutein™ (Grifols, Clayton, NC), Buminate™ (Baxatla, Inc., Bannockburn, IL), Flexbumin™ (Baxatla, Inc., Bannockburn, IL), and Plasbumin™ (Grifols, Clayton, NC).

[0161] Human plasma protein components (PPF)

[0162] According to the U.S. Food and Drug Administration (“FDA”), “Plasma Protein Component (Human)” or PPF is the correct name for this product, defined as “a sterile protein solution of albumin and globulin extracted from human plasma” (Federal Regulation “CFR” 21 CFR 640.90, incorporated herein by reference). The raw material source for PPF is whole blood recycled plasma prepared in accordance with 21 CFR 640.1–640.5 (incorporated herein by reference), or raw material plasma prepared in accordance with 21 CFR 640.60–640.76 (incorporated herein by reference).

[0163] PPF must be tested to determine whether it conforms to the following standards as specified in 21 CFR 640.92 (incorporated herein by reference):

[0164] (a) The final product should be a solution with a protein concentration of 5.0% + / - 0.30%; and

[0165] (b) The total protein in the final product shall contain at least 83% albumin and no more than 17% globulin. Gamma globulin shall not exceed 1% of the total protein. Protein composition shall be determined using a method approved by the Director of the Centre for Biologics Evaluation and Research, FDA, and each manufacturer’s method shall be separately approved.

[0166] As used herein, “plasma protein fraction” or “PPF” refers to a sterile protein solution derived from human plasma, consisting of albumin and globulins, which, as determined by electrophoresis, contains at least 83% albumin, no more than 17% globulins (including α1-globulin, α2-globulin, β-globulin, and γ-globulin) and other plasma proteins, and no more than 1% γ-globulin. (Hink, JH, Jr., et al., Preparation and Properties of a Heat-Treated Human Plasma Protein Fraction, VOX SANGUINIS 2(174) (1957)). PPF can also refer to a solid form, which has a similar composition when suspended in a solvent. The total globulin fraction can be determined by subtracting albumin from the total protein. (Busher, J., Serum Albumin and Globulin, CLINICAL METHODS: THE HISTORY, PHYSICAL, AND LABORATORY EXAMINATIONS, Chapter 10, Walker HK, Hall WD, Hurst JD, editors. (1990)).

[0167] Human albumin (HAS)

[0168] According to the FDA, “Albumin (Human)” (also referred to herein as “HAS”) is the correct name for this product, defined as “a sterile solution of albumin extracted from human plasma” (Federal Regulation “CFR” 21 CFR 640.80, incorporated herein by reference). Albumin (Human) is derived from whole blood recycled plasma prepared in accordance with 21 CFR 640.1–640.5 (incorporated herein by reference), or from raw plasma prepared in accordance with 21 CFR 640.60–640.76 (incorporated herein by reference). Other requirements for Albumin (Human) are listed in 21 CFR 640.80–640.84 (incorporated herein by reference).

[0169] Albumin (human) must be tested to determine whether it meets the following criteria as specified in 21 CFR 640.82:

[0170] (a) Protein concentration. The final product shall be a protein solution with a protein concentration of 4.0% + / - 0.25%; 5.0% + / - 0.30%; 20.0% + / - 1.2%; and 25.0% + / - 1.5%.

[0171] (b) Protein composition. The total protein in the final product should contain at least 96% albumin, determined by a method approved by the Director of the Centre for Biological Evaluation and Research, FDA. Each manufacturer’s method requires separate approval.

[0172] As used herein, “albumin (human)” or “HAS” refers to a sterile protein solution derived from human plasma, consisting of albumin and globulins, with an albumin content of at least 95% and globulins (including α1-globulin, α2-globulin, β-globulin, and γ-globulin) and other plasma proteins not exceeding 5%. HAS can also refer to a solid form, having a similar composition when suspended in a solvent. The total globulin fraction can be determined by subtracting albumin from the total protein content.

[0173] Those skilled in the art will recognize that the PPF and HAS fractions can also be lyophilized or exist in other solid forms. Such formulations, with the addition of appropriate additives, can be used to manufacture, for example, tablets, powders, granules, or capsules. The solid form can be formulated into an injection formulation by dissolving, suspending, or emulsifying in an aqueous solution; and, if desired, conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, and preservatives can be added.

[0174] F. Indications

[0175] The methods and compositions of this invention can be used to treat cognitive impairment, such as cognitive impairment caused by neuroinflammation. In some cases, cognitive impairment is age-related, such as age-related impairment of an individual's cognitive abilities.

[0176] Non-limiting examples of such cognitive impairment include age-related dementia, immune diseases, and physical or functional decline. Individuals who have or are suspected of having cognitive impairment and who may benefit from the treatments disclosed herein include individuals aged approximately 50 years or older, such as 60 years or older, 70 years or older, 80 years or older, 90 years or older, and 100 years or older, i.e., individuals aged approximately 50 to 100 years, such as 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or approximately 100 years, and who have cognitive impairment associated with natural aging. Individuals with age-related cognitive impairment, such as mild cognitive impairment (MCI); and individuals aged approximately 50 years or older, such as 60 years or older, 70 years or older, 80 years or older, 90 years or older, and generally not exceeding 100 years of age, i.e., individuals aged approximately 50 to 90 years, such as 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or approximately 100 years of age, who have not yet begun to show symptoms of cognitive impairment. Examples of cognitive impairment / indications that may be caused by natural aging include the following:

[0177] 1. Mild cognitive impairment (MCI)

[0178] Mild cognitive impairment is a mild cognitive disorder manifested as problems with memory or other mental functions, such as planning, following instructions, or decision-making. These problems worsen over time, but overall mental function and daily activities are not impaired. Therefore, although significant neuronal death does not typically occur, neurons in the aging brain are susceptible to sublethal age-related structural, synaptic integrity, and synaptic molecular processing alterations, all of which impair cognitive function. Individuals with or suspected of having age-related cognitive impairment will benefit from treatment using the methods disclosed in this invention.

[0179] This article also envisions individuals of any age with cognitive impairment due to age-related diseases; and individuals of any age diagnosed with an age-related disorder that is typically accompanied by cognitive impairment. Examples of such age-related disorders include the following:

[0180] 2. Alzheimer's disease (AD)

[0181] Alzheimer's disease is a progressive, unstoppable loss of cognitive function, associated with excessive senile plaques in the cerebral cortex and subcortical gray matter. These plaques also contain beta-amyloid protein and neurofibrillary tangles composed of tau protein. The most common form affects people over 60 years of age, and its incidence increases with age. Alzheimer's disease accounts for more than 65% of dementia cases in the elderly.

[0182] The cause of Alzheimer's disease (AD) is unclear. The disease has a familial inheritance pattern in approximately 15% to 20% of cases. The remaining so-called sporadic cases have some genetic determinants. The disease exhibits an autosomal dominant inheritance pattern in most early-onset cases and some late-onset cases, but with variable penetrance in later life. Environmental factors are a focus of active research.

[0183] During the disease process, synapses and terminal neurons are lost in the cerebral cortex, hippocampus, and subcortical structures (including selective cell loss in the basal ganglia of Minert), locus coeruleus, and dorsal raphe nucleus. Glucose utilization and perfusion are reduced in certain brain regions (parietal and temporal lobes in early-stage disease, and prefrontal cortex in late-stage disease). Neuroinflammatory plaques or senile plaques (composed of neurites, astrocytes, and glial cells surrounding an amyloid core) and neurofibrillary tangles (composed of paired spiral filaments) play a role in the pathogenesis of Alzheimer's disease (AD). While senile plaques and neurofibrillary tangles can also appear during normal aging, they are more prevalent in AD patients.

[0184] 3. Parkinson's disease

[0185] Parkinson's disease (PD) PD is an idiopathic, slowly progressive, degenerative central nervous system disease characterized by bradykinesia (reduced movement and activity), rigidity, resting tremor (dystonia), muscle freezing, and postural instability. Initially considered primarily a motor disorder, it is now recognized that PD can also cause depression and mood changes. PD can also affect cognition, behavior, sleep, autonomic function, and sensory function. The most common cognitive impairments include attention and concentration, working memory, executive function, language production, and visuospatial function. A characteristic feature of PD is that symptoms associated with decreased motor function usually precede those associated with cognitive impairment, which aids in diagnosis.

[0186] In primary Parkinson's disease, pigmentary neurons in the substantia nigra, locus coeruleus, and other brainstem dopaminergic cell populations degenerate. The cause is unknown. The loss of substantia nigra neurons (which project to the caudate nucleus and putamen) leads to the depletion of the neurotransmitter dopamine in these areas. Onset typically occurs after age 40, with an increased incidence in the elderly.

[0187] Approximately 60,000 new cases of Parkinson's disease are diagnosed in the United States each year, currently affecting about one million Americans. Although PD itself is not fatal, its complications are the 14th leading cause of death in the United States. Currently, there is no cure for PD; treatment usually aims to control symptoms, and surgery is only used in advanced, severe cases.

[0188] Treatment options for Parkinson's disease (PD) include the use of medications to help manage motor deficits. These options increase or replace the neurotransmitter dopamine, which is present in lower concentrations in the brains of PD patients. Such medications include: carbidopa / levodopa (which produces more dopamine in the brain); apomorphine, pramipexole, ropinirole, and rotigotine (dopamine agonists); selegiline and rasagiline (MAO-B inhibitors that prevent dopamine breakdown); entacapone and tocapone (catechol-O-methyltransferase [COMT] inhibitors, making more levodopa available in the brain); benzalkonium chloride and trihexyphenidyl (anticholinergics); and amantadine (to control tremors and rigidity). Exercise / physical therapy is also commonly used to help maintain physical and mental function.

[0189] However, current treatments only address the symptoms of PD, failing to cure the disease or halt its progression. Furthermore, current medications often become ineffective in later stages of PD. The most commonly used drug, levodopa, typically produces adverse effects within 5 to 10 years of starting treatment. These adverse effects can be severe, leading to motor fluctuations and unpredictable fluctuations in motor control during treatment, as well as tics / convulsions (motor disorders), which are difficult to manage and can even be as disabling as the symptoms of PD itself. Therefore, new therapies with novel mechanisms of action, which can be administered alone or in combination with current PD medications, remain in demand.

[0190] 4. Parkinson's syndrome

[0191] Secondary Parkinsonism (also known as atypical Parkinson's disease or Parkinson's plus syndrome) is caused by the loss or blockage of dopamine activity in the basal ganglia due to other idiopathic degenerative diseases, medications, or exogenous toxins. The most common cause of secondary Parkinsonism is the ingestion of antipsychotic drugs or reserpine, which induce Parkinsonism by blocking dopamine receptors. Less common causes include carbon monoxide or manganese poisoning, hydrocephalus, structural lesions (tumors or infarctions affecting the midbrain or basal ganglia), subdural hematoma, and degenerative diseases, including substantia nigra-striatal degeneration. Certain diseases, such as progressive supranuclear palsy (PSP), multiple system atrophy (MSA), corticobasal degeneration (CBD), and Lewy body dementia (DLB), may present with Parkinsonism symptoms before the major symptoms required for a specific diagnosis appear, and may therefore be labeled as "Parkinsonism syndrome."

[0192] 5. Frontotemporal dementia

[0193] Frontotemporal dementia (FTD) is a disease caused by progressive degeneration of the frontal lobe of the brain. Over time, this degeneration may extend to the temporal lobe. FTD is the second most common dementia after Alzheimer's disease (AD), accounting for 20% of Alzheimer's cases. Symptoms are divided into three groups based on the function of the affected frontal and temporal lobes:

[0194] Behavioral variant FTD (bvFTD) presents with symptoms including somnolence and loss of initiative on one hand, and disinhibition on the other; progressive nonfluent aphasia (PNFA) is characterized by interrupted speech fluency due to articulation difficulties, phonological and / or syntactic errors, but word comprehension is preserved; and semantic dementia (SD) is characterized by fluent speech with normal phonology and syntax, but gradually increasing difficulty in naming and word comprehension. Other common cognitive symptoms in all FTD patients include impaired executive function and concentration. Other cognitive abilities, including perception, spatial skills, memory, and application abilities, are usually intact. FTD can be diagnosed by observing atrophy of the frontal lobe and / or anterior temporal lobe on structural MRI scans.

[0195] There are various forms of frontotemporal dementia, any of which can be treated or prevented using the methods and compositions of this invention. For example, one form of frontotemporal dementia is semantic dementia (SD). SD is characterized by loss of semantic memory in both verbal and nonverbal domains. Patients with SD typically complain of difficulty finding words. Clinical signs include fluent aphasia, naming difficulties, impaired understanding of word meaning, and associative visual agnosia (inability to match semantically relevant pictures or objects). As the disease progresses, behavioral and personality changes often occur, similar to those observed in frontotemporal dementia, although there are also case descriptions of “simple” semantic dementia, with fewer late-stage behavioral symptoms. Structural MRI imaging shows a characteristic pattern of temporal lobe atrophy (primarily on the left side), with greater involvement of the inferior temporal lobe than the superior temporal lobe, and greater atrophy of the anterior temporal lobe than the posterior temporal lobe.

[0196] Another example of frontotemporal dementia is Pick's disease (PiD, also known as PcD). The defining characteristic of this disease is the accumulation of tau protein in neurons, forming silver-stained spherical aggregates called "Pick bodies." Symptoms include loss of speech (aphasia) and dementia. Patients with impaired orbitofrontal lobe function may become aggressive and exhibit socially inappropriate behaviors. They may steal or display compulsive or repetitive stereotyped behaviors. Patients with impaired dorsolateral nucleus or dorsolateral frontal lobe function may exhibit apathy, emotional blunting, or decreased initiative. Patients may exhibit a lack of self-monitoring, abnormal self-awareness, and an inability to understand meaning. Patients with bilateral posterolateral orbitofrontal cortex and right anterior insula gray matter loss may exhibit changes in eating behavior, such as morbid cravings for sweets. Patients with more pronounced focal gray matter loss in the anterolateral orbitofrontal cortex may develop hyperphagia. Although some symptoms may initially be relieved, the disease progresses, and patients typically die within two to ten years.

[0197] 6. Huntington's disease

[0198] Huntington's disease (HD) is a hereditary, progressive neurodegenerative disorder characterized by the development of mood, behavior, and psychotic abnormalities; loss of intellectual or cognitive function; and motor abnormalities (motor disorders). Typical signs of HD include the development of chorea—involuntary, rapid, irregular, and agitated movements that may affect the face, arms, legs, or trunk—and cognitive decline, including a gradual loss of cognitive processing abilities and acquired intellectual capacity. Impaired memory, abstract thinking, and judgment may occur; disorientation (misperception of time, place, or identity); increased agitation; and personality changes (depersonalization). While symptoms typically become apparent in one's forties or fifties, the age of onset is variable, ranging from early childhood to late adulthood (e.g., in one's seventies or eighties).

[0199] HD is inherited in families as an autosomal dominant trait. The disorder is caused by an abnormally long sequence or "duplication" of a gene encoding an instruction on chromosome 4 (4p16.3). The progressive loss of neurological function associated with HD is due to the loss of neurons in certain areas of the brain, including the basal ganglia and cerebral cortex.

[0200] 7. Amyotrophic Lateral Sclerosis (ALS)

[0201] Amyotrophic lateral sclerosis (ALS) is a rapidly progressive and inevitably fatal neurological disease that attacks motor neurons. Signs of muscle weakness and atrophy, as well as anterior horn cell dysfunction, most commonly appear in the hands initially, and less frequently in the feet. The site of onset is random, and progression is asymmetrical. Spasticity is common and may precede weakness. Few patients survive 30 years; 50% die within 3 years of onset, 20% survive 5 years, and 10% survive 10 years.

[0202] Diagnostic features include onset in middle or late adulthood and progressive, generalized motor involvement without sensory abnormalities. Nerve conduction velocity remains normal until the late stages of the disease. Recent studies have also documented cognitive impairment, particularly declines in immediate verbal memory, visual memory, language, and executive function.

[0203] Reports indicate that even neurons that appear normal in ALS patients exhibit reduced cell body area, synapse number, and total synaptic length. It has been proposed that persistent synaptic loss may lead to dysfunction when the plasticity of active areas reaches its limit. Promoting the formation of new synapses or preventing synaptic loss may help maintain neuronal function in these patients.

[0204] 8. Multiple sclerosis

[0205] Multiple sclerosis (MS) is characterized by a variety of symptoms and signs of central nervous system dysfunction, with remissions and relapses. The most common initial symptoms are sensory disturbances in one or more limbs, trunk, or one side of the face; weakness or clumsiness in the legs or hands; or visual disturbances such as partial blindness and monocular pain (retrobulbar optic neuritis), blurred vision, or scotomas. Common cognitive impairments include impaired memory (acquiring, retaining, and retrieving new information), attention and concentration (especially distributive attention), information processing, executive function, visuospatial function, and verbal fluency. Common early symptoms include oculomotor palsy leading to diplopia (diplopia), transient weakness in one or more limbs, mild stiffness or unusual fatigue of limbs, mild gait disturbances, bladder control difficulties, dizziness, and mild mood disturbances; all of these indicate diffuse involvement of the central nervous system and often appear months or years before the disease is recognized. Overheating may exacerbate symptoms and signs.

[0206] The course of the disease is highly variable and unpredictable, and in most patients it is remission-prone. Initially, there may be remission periods of several months or years between attacks, especially when the disease begins with retrobulbar optic neuritis. However, some patients experience frequent attacks and rapid disability; in a minority of patients, the disease course may progress rapidly.

[0207] 9. glaucoma

[0208] Glaucoma is a common neurodegenerative disease that affects retinal ganglion cells (RGCs). Evidence supports a compartmentalized degenerative process in synapses and dendrites, including retinal ganglion cells. Recent evidence also suggests a correlation between cognitive impairment and glaucoma in older adults (Yochim BP et al., Prevalence of cognitive impairment, depression, and anxiety symptoms among older adults with glaucoma. J Glaucoma. 2012;21(4):250-254).

[0209] 10. Myotonic dystrophy

[0210] Myotonic dystrophy (DM) is an autosomal dominant multisystem disorder characterized by dystrophic muscle weakness and rigidity. The molecular defect is an amplification of the trinucleotide (CTG) repeat sequence in the 3' untranslated region of the tonicin kinase gene on chromosome 19q. Symptoms can occur at any age and range in clinical severity. Rigidity is prominent in the muscles of the hands, and ptosis is common even in mild cases. In severe cases, significant peripheral muscle weakness is present, often accompanied by cataracts, premature baldness, axe-like facial features, cardiac arrhythmias, testicular atrophy, and endocrine abnormalities (such as diabetes). Intellectual disability is common in the severe congenital form, while in the milder adult form of the disease, age-related decline in frontal and temporal lobe cognitive function, particularly language and executive functions, is observed. Severely affected individuals die in their early 50s.

[0211] 11. dementia

[0212] Dementia describes a class of diseases that severely affect thinking and social abilities to the point of interfering with daily functioning. Besides dementia observed later in the age-related diseases mentioned above, other examples of dementia include vascular dementia and Lewy body dementia, as described below.

[0213] In vascular dementia, or "multiple infarct dementia," cognitive impairment is caused by problems with blood supply to the brain, often resulting from a series of minor strokes, or sometimes, a major stroke followed by other smaller strokes. Vascular lesions can be diffuse (e.g., small vessel disease) or focal lesions, or a combination of both. Following an acute cerebrovascular event, patients with vascular dementia present with acute or subacute cognitive impairment, followed by a progressive decline in cognitive abilities. The cognitive impairment is similar to that observed in Alzheimer's disease, including impairments in language, memory, complex visual processing, or executive function, although the related changes in the brain are not due to AD pathology but rather to a chronic reduction in cerebral blood flow that ultimately leads to dementia. Single-photon emission computed tomography (SPECT) and positron emission tomography (PET) neuroimaging, combined with mental status examinations, can be used to confirm the diagnosis of multiple infarct dementia.

[0214] Lewy body dementia (DLB, also known by several other names including Lewy body dementia, diffuse Lewy body disease, cortical Lewy body disease, and Lewy body-type Alzheimer's disease) is a type of dementia characterized anatomically by the presence of Lewy bodies (clumps of α-synuclein and ubiquitin proteins) in neurons, detectable in post-mortem brain histology. The main characteristic of Lewy body dementia is cognitive decline, particularly executive function. Alertness and short-term memory may fluctuate.

[0215] Vivid and detailed visual hallucinations, whether persistent or recurring, are often an early diagnostic symptom. DLB is frequently confused with Alzheimer's disease and / or vascular dementia in its early stages. While Alzheimer's disease typically has a slow onset, DLB often has a rapid or acute onset. DLB symptoms also include motor symptoms similar to those of Parkinson's disease. The difference between DLB and dementia, which sometimes occurs in Parkinson's disease, lies in the timeframe of the dementia symptoms relative to the Parkinson's symptoms. When dementia begins more than a year after the onset of Parkinson's disease, the diagnosis is Parkinson's disease with dementia (POD). When cognitive symptoms occur simultaneously with Parkinson's symptoms or begin within a year of the onset of Parkinson's symptoms, the diagnosis is DLB.

[0216] 12. CADASIL

[0217] Autosomal dominant cerebral arteriosclerosis with subcortical infarction and leukoencephalopathy (CADASIL) is a genetic disorder associated with mutations in the NOTCH3 gene (Locatelli M, et al., Front. Pharmacol. 11:321(2020)). It typically occurs in middle-aged adults and manifests as cognitive impairment leading to dementia and disability (ibid.). Other manifestations include mood disorders, migraines with aura, and recurrent strokes. Effective treatments have been difficult to find because the pathogenesis of the disease remains unclear (ibid.). CADASIL is the most common inherited subcortical vascular dementia (Kalimo H, et al., Future Neurology, 3(6) (2008)).

[0218] CADASIL is characterized by four main common symptoms: migraine with aura, recurrent ischemic stroke, psychiatric disorders, and cognitive decline. The first symptom is usually the first symptom, occurring in 20% to 40% of patients. The second symptom occurs in 60% to 85% of symptomatic individuals. The third symptom, psychiatric disorders, occurs in 25% to 30% of patients and manifests as moderate / major depression, bipolar disorder, panic disorder, schizophrenia, and emotional blunting. Cognitive impairment occurs in 60% of patients, becoming clinically detectable between the ages of 35 and 50, and gradually worsening with age (ibid.). In younger patients, attention, memory, and executive function impairments are predominant (Buffon F, et al., J NeurolNeurosurg Psychiatry 77(2):175-80 (2006)). Visuospatial and reasoning abilities deteriorate with age, primarily after age 60. Dementia occurs in 25% of patients, with 75% of these being over 60 years of age. However, the frequency of ischemic attacks is not related to dementia. (Ibid.)

[0219] CADASIL is a progressive and fatal disease. To date, there are no disease-modifying treatments. (Locatelli et al., ibid.). Symptomatic treatment is the only option for clinicians, based on routine clinical practice, such as: acetazolamide or sodium valproate for migraines; daily aspirin to reduce the risk of heart attack or stroke; and supportive care for cognitive loss. Notably, no medication has yet been clearly shown to be beneficial for CADASIL-related cognitive loss. (ibid.). Interventions that have been studied but failed include donepezil (for improving cognitive impairment in Alzheimer's disease), galantamine (an acetylcholinesterase inhibitor for treating cognitive impairment in Alzheimer's disease), and levodopa (for Alzheimer's and Parkinson's diseases).

[0220] 13. Progressive supranuclear palsy

[0221] Progressive supranuclear palsy (PSP) is a brain disorder that causes severe and progressively worsening problems with gait and balance control, complex eye movements, and thinking. One of the hallmarks of the disease is the inability to properly align the eyes, due to damage to areas of the brain that coordinate eye movements. Some individuals describe this effect as blurring. Affected individuals often exhibit mood and behavioral changes, including depression and emotional blunting, as well as progressive mild dementia. The long name of the disease indicates that it begins slowly and worsens over time (progressive) and causes weakness (paralysis) by damaging parts of pea-sized structures in the brain that control eye movements, called nuclei. PSP was first described as a distinct disease in 1964 when three scientists published a paper differentiating it from Parkinson's disease. It is sometimes referred to as the Steele-Richardson-Olszewski syndrome, reflecting the combined name given by the scientists who defined the disease. Although PSP progresses gradually, no one dies from PSP itself.

[0222] 14. Ataxia

[0223] Ataxia is a condition characterized by coordination problems caused by impaired portions of the nervous system that control movement and balance. Ataxia can affect the fingers, hands, arms, legs, body, speech, and eye movements. The term "ataxia" is commonly used to describe symptoms of poor coordination that may be associated with infection, injury, other diseases, or degenerative changes in the central nervous system. Ataxia is also used to refer to a specific group of neurodegenerative disorders called hereditary ataxia and sporadic ataxia, which is the primary focus of the Ataxia Foundation in the United States.

[0224] 15. Multiple system atrophy

[0225] Multiple system atrophy (MSA) is a degenerative neurological disorder. MSA is associated with the degeneration of nerve cells in specific areas of the brain. This cellular degeneration leads to problems with movement, balance, and other autonomic bodily functions, such as bladder control or blood pressure regulation.

[0226] The cause of MSA is unknown, and no specific risk factors have been identified. Approximately 55% of cases occur in men, with a typical age of onset in the late 1950s to early 1960s. MSA often presents with some of the same symptoms as Parkinson's disease. However, MSA patients typically have little or no response to dopamine medications used to treat Parkinson's disease.

[0227] 16. weak

[0228] Frailty is an aging syndrome characterized by functional and physical decline, including decreased mobility, muscle weakness, slowness, poor endurance, reduced physical activity, malnutrition, and involuntary weight loss. This decline is often accompanied by and is a consequence of diseases such as cognitive impairment and cancer. However, frailty can occur even in the absence of disease. Individuals with frailty syndrome have an increased risk of fractures, accidental falls, disability, comorbidities, and premature death (C. Buigues, et al., Effect of a Prebiotic Formulation on Frailty Syndrome: A Randomized, Double-Blind Clinical Trial, Int. J. Mol. Sci. 2016, 17, 932). Furthermore, individuals with frailty syndrome have a higher incidence of increased healthcare expenditures (ibid.).

[0229] Common symptoms of frailty syndrome can be identified by certain types of tests. For example, involuntary weight loss involves a loss of at least 10 pounds or more than 5% of body weight in the previous year; muscle weakness can be identified by the lowest 20% of grip strength at baseline (adjusted for sex and BMI); bradykinesia can be identified based on the time required to walk 15 feet; poor endurance can be identified based on an individual's self-reported fatigue; and low physical activity can be measured using standardized questionnaires. (Z. Palace et al., The Frailty Syndrome, Today's Geriatric Medicine 7(1), at18 (2014)).

[0230] 17. Neuromyelitis optica spectrum disorders

[0231] Neuromyelitis optica spectrum disorder (NMOSD), also known as Devic disease, is a rare inflammatory disease of the central nervous system. It is characterized by optic neuritis (inflammation of the optic nerve) and myelitis (inflammation of the spinal cord). Patients typically experience recurrent episodes of inflammation interspersed with remission. The disease is believed to be caused by autoantibodies that typically target myelin oligodendrocyte glycoprotein (MOG-IgG) or aquaporin 4 (AQP4-IgG), leading to demyelination of the optic nerve and spinal cord, and axonal damage.

[0232] 18. Postoperative cognitive impairment

[0233] Postoperative cognitive decline occurs after anesthesia and surgery. This is common in patients over 60 years of age and is diagnosed through preoperative and postoperative cognitive tests. Patients typically present with memory impairment, delirium, and impaired performance on intellectual tasks.

[0234] 19. Chronic traumatic encephalopathy

[0235] Chronic traumatic encephalopathy (CTE) is a neurodegenerative brain disorder most commonly seen in athletes, veterans, or others with a history of recurrent head trauma. It is one of many tau protein disorders characterized by an excess of tau protein in the brain, leading to neuronal loss. Symptoms include memory loss, mood or personality changes, confusion, impaired judgment, impulse control disorders, aggression, and depression.

[0236] 20. Traumatic brain injury

[0237] Traumatic brain injury (TBI) is caused by a severe impact to the head or body. It can also be caused by an object penetrating brain tissue during the injury. It leads to bleeding, tissue tears, physical damage to brain cells, and cell death. Physical symptoms vary, but include loss of consciousness, headache, nausea, extreme fatigue, speech difficulties, sleep problems, dizziness, blurred vision, sensitivity to light or sound, memory loss, and attention problems.

[0238] In some embodiments, the methods and compositions of the present invention can be used to slow the progression of age-related cognitive impairment. In other words, after receiving treatment with the methods disclosed in the present invention, the rate of decline in cognitive impairment will be slower than before treatment or without treatment with the methods disclosed in the present invention. In some such cases, the treatment methods of the present invention include measuring the progression of cognitive impairment after treatment and determining the slowing of the decline. In some such cases, this is determined by comparison with a reference, for example, by comparing with the individual's pre-treatment rate of decline, for example, by measuring cognitive ability at two or more time points before application of the blood product of the present invention.

[0239] The methods and compositions of this invention are also used to stabilize the cognitive abilities of individuals, such as those with age-related cognitive impairment or those suspected of having age-related cognitive decline. For example, an individual may exhibit certain age-related cognitive impairments, and after receiving treatment with the methods disclosed in this invention, the progression of cognitive impairment observed before treatment will cease.

[0240] As another example, an individual may be at risk of developing age-related cognitive decline (e.g., the individual may be 50 years of age or older, or may have been diagnosed with an age-related disease), and after receiving treatment with the method disclosed in this invention, the individual's cognitive abilities remain substantially unchanged compared to before treatment, i.e., no cognitive decline is detected.

[0241] The methods and compositions of the present invention are also used to reduce cognitive impairment in individuals suffering from age-related damage. In other words, the cognitive abilities of affected individuals are improved after treatment with the methods of the present invention. For example, an individual's cognitive abilities increase relative to pre-treatment cognitive abilities after treatment with the methods of the present invention, for example, by 2 times or more, 5 times or more, 10 times or more, 15 times or more, 20 times or more, 30 times or more, or 40 times or more, including 50 times or more, 60 times or more, 70 times or more, 80 times or more, 90 times or more, or 100 times or more.

[0242] In some cases, treatment with the methods and compositions of the present invention restores the cognitive abilities of individuals suffering from age-related cognitive impairment to the level they would have had at approximately 40 years of age or younger. In other words, cognitive or motor impairment is eliminated.

[0243] G. Methods for monitoring improvement

[0244] In some cases, among various methods for monitoring disease progression and improvement in cognitive impairment, the following types of assessments are used alone or in combination for subjects with cognitive impairment. These methods are presented as examples and are not limited to. Any convenient method for monitoring disease can be used to practice this invention as needed. These methods also fall within the scope of the methods of this invention.

[0245] i. General understanding

[0246] Some embodiments of the method of the present invention also include methods for monitoring the effects of a drug or treatment used to treat cognitive impairment (e.g., age-related cognitive impairment) on the cognitive impairment of a subject. Some such methods include comparing cognitive function before and after treatment. Methods for assessing cognitive function are well known in the art. For example, but not limited to, methods may include assessing cognitive function based on medical history, family history, physical and neurological examinations performed by a clinician specializing in cognitive function, laboratory tests, and neuropsychological assessments. Other embodiments contemplated by the present invention include: assessment of consciousness, such as using the Glasgow Coma Scale (EMV); examination of mental status, including the Mini-Mental State Examination (AMTS) or the Mini-Mental State Examination (MMSE) (Folstein et al., J. Psychiatr. Res 1975; 12:1289-198); overall assessment of higher functions; and estimation of intracranial pressure, such as by funduscopy. In one implementation, monitoring the effects on cognitive impairment (e.g., age-related cognitive impairment) includes examining improvements in scores of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 using the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-COG).

[0247] In one implementation, an examination of the peripheral nervous system can be used to assess cognitive function, including any of the following: olfaction, visual field and visual acuity, eye movements and pupils (sympathetic and parasympathetic), facial sensory function, facial and shoulder girdle muscle strength, hearing, taste, pharyngeal motor and reflexes, and tongue movements, which can be tested individually (e.g., visual acuity can be tested using the Snellen visual acuity chart; reflexes are tested using a reflex hammer, including the masseter reflex, biceps and triceps tendon reflexes, patellar reflex, ankle reflex, and plantar reflex (i.e., Babinski sign); muscle strength is typically assessed using an MRC grading from 1 to 5; and signs of muscle tone and rigidity).

[0248] Some embodiments of the methods disclosed herein also include monitoring the effects of treatment for cognitive impairment on motor function before and after treatment. Some such methods include assessing motor function in PD patients treated for cognitive impairment according to the methods disclosed herein. Methods for assessing motor function are well known in the art. For example, but not limited to, these methods may include assessing motor function based on medical history, family history, physical and neurological examinations performed by a clinician specializing in neurodegenerative and motor disorders, laboratory tests, and neurodegenerative assessments. Other embodiments contemplated by the invention include the use of rating scales discussed below.

[0249] Several rating scales have been used to assess the progression of PD. The most widely used scales include the Unified Parkinson's Disease Rating Scale (UPDRS, introduced in 1987) (J. Rehabil Res. Dev., 2012 49(8): 1269-76) and the Hoehn and Yahr Scale (Neruology, 1967 17(5): 427-42). Other scales include the Movement Disorders Society (MDS) updated UPDRS scale (MDS-UPDRS) and the Schwab and England Activities of Daily Living (ADL) Scale.

[0250] The UPDRS scale assesses 31 items, which are divided into three subscales: (1) mental activity, behavior, and mood; (2) activities of daily living; and (3) motor examination. The Hoehn and Yahr scale divides PD into five stages with well-defined substages: 0 – no signs of disease; 1 – symptoms only on one side; 1.5 – symptoms on one side, but also involving the neck and spine; 2 – symptoms on both sides, without balance impairment; 2.5 – mild symptoms on both sides, recoverable on a “pull” test; 3 – balance impairment, mild to moderate; 4 – severe disability, but able to walk or stand independently; 5 – requires a wheelchair or is bedridden and unable to care for oneself. The Schwab and England scale divides PD into several percentages (from 100% – completely independent to 10% – completely dependent).

[0251] General motor function can be assessed using widely used scales, including the General Motor Function Scale (GMF). This test comprises three components: dependence, pain, and insecurity. (Aberg AC, et al., (2003) Disabil. Rehabil. 2003 May 6;25(9):462-72.). Motor function can also be assessed using home monitoring or wearable sensors. For example: gait (speed of movement, variability, leg rigidity) can be sensed using accelerometers; posture (trunk tilt) using gyroscopes; leg movements using accelerometers; hand movements using accelerometers and gyroscopes; tremor (amplitude, frequency, duration, asymmetry) using accelerometers; falls using accelerometers; gait freeze using accelerometers; movement disorders using accelerometers, gyroscopes, and inertial sensors; bradykinesia (duration and frequency) using accelerometers and gyroscopes; and aphasia (pitch) using microphones. (Pastorino M, et al., Journal of Physics: Conference Series 450 (2013)012055).

[0252] Some embodiments of the methods disclosed herein also include monitoring the effect of treatment for cognitive impairment on the progression or improvement of neurodegenerative disease. Some such methods include assessing the progression or improvement of neurodegenerative disease in subjects with neurodegenerative diseases (such as MS, HD, ALS, glaucoma, PSP) who are treated for cognitive impairment according to the methods disclosed herein.

[0253] Methods for monitoring the progression or improvement of neurodegenerative diseases are well known to those skilled in the art. For example, and not limitingly, monitoring can be performed using techniques such as: cerebrospinal fluid (CSF) monitoring; magnetic resonance imaging (MRI) to detect the development of lesions and demyelinating plaques; evoked potential studies; and gait monitoring.

[0254] CSF analysis can be performed, for example, by lumbar puncture to obtain pressure, appearance, and CSF composition. Normal values ​​typically range as follows: pressure (70 mm H2O to 180 mm H2O); appearance: clear and colorless; total protein (15 mg / 100 mL to 60 mg / 100 mL); IgG as a percentage of total protein: 3% to 12%; glucose (50 mg / 100 mL to 80 mg / 100 mL); cell count: 0 to 5 white blood cells, no red blood cells; chloride (110 mEq / L to 125 mEq / L). Abnormal results may indicate the presence or progression to MS.

[0255] MRI is another technique that can be used to monitor disease progression and improvement. Typical criteria for monitoring MS using MRI include patchy abnormal white matter areas in the cerebral hemispheres and periventricular regions, lesions in the cerebellum and / or brainstem, and cervical or thoracic spinal cord regions.

[0256] Evoked potentials can be used to monitor the progression and improvement of metastatic disorders (MS) in subjects. Evoked potentials measure a slowing of electrical impulses, such as visual evoked potentials (VER), brainstem auditory evoked potentials (BAER), and somatosensory evoked potentials (SSER). Abnormal responses help indicate a slowing of conduction velocity in central sensory pathways.

[0257] Gait monitoring can also be used to monitor disease progression and improvement in MS subjects. MS is often accompanied by impaired mobility and abnormal gait, partly due to fatigue. Monitoring can be performed using a motion monitoring device worn by the subject. (Moon, Y., et al., Monitoring gait in multiple sclerosis with novel wearable motion sensors, PLOS One, 12(2):e0171346 (2017)).

[0258] This disclosure also envisions objects that treat or improve degenerative or damaged neuropathy, which may manifest as cognitive or motor dysfunction or as neuroinflammation.

[0259] Embodiments of the present invention also envision determining neurogenesis levels before, during, and / or after treatments according to this disclosure, such as plasma exchange therapy. Non-invasive techniques for assessing neurogenesis have been reported (Tamura Y. et al., J. Neurosci. (2016) 36(31):8123-31). Positron emission tomography (PET) combined with the tracer [18F]FLT and the BBB transporter inhibitor probenecid allows the tracer to accumulate in neurogenesis regions of the brain. This imaging can assess neurogenesis in patients undergoing treatment for neurodegenerative diseases.

[0260] H. Sports

[0261] Exercise can be characterized by aerobic or anaerobic activity, and can involve high-calorie-burning and moderate-calorie-burning activities. Exercise may involve strength training (e.g., weight training or isometric exercises). Exercise may also involve, for example, running, cycling, walking, dancing, walking, swimming, yoga, tai chi, balance exercises, leg bends, rope skipping, surfing, rowing, rotating or bending arms or legs, gardening, cleaning, active games (such as bowling), aerobic exercise, Pilates, and martial arts.

[0262] The exercise program may include performing one exercise at a certain frequency, or a combination of exercises at a certain frequency. The frequency may be once, twice, three times, four times, five times, six times, or seven times per week. The frequency may vary from week to week. The exercise program may be at the same intensity and / or frequency level as the subject before applying the composition of the present invention. The exercise program may also have a higher intensity and / or frequency than the level practiced by the subject before applying the therapy of the present invention. The exercise program may be advised or prescribed by a health or fitness professional, or the exercise program may be initiated by the subject themselves.

[0263] I. Reagents, Kits, and Equipment

[0264] Some aspects of the present invention provide a kit comprising reagents for measuring the amount of one or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in a sample.

[0265] As stated above, the detection of one or more target proteins can be performed by any suitable method, such as qualitative or quantitative methods. Therefore, the kits disclosed herein can be designed for the qualitative or quantitative detection of one or more target proteins.

[0266] Therefore, in some cases, the kits disclosed herein provide reagents and / or devices for determining the sequence of target proteins. Certain such reagents and / or devices can be used for qualitative target protein analysis using mass spectrometry. These reagents and / or devices include reagents for lysing proteins in a sample, such as enzymes for digesting proteins, like trypsin; various solvents, such as water, methanol, acetonitrile, chloroform, tetrahydrofuran, and dichloromethane; and various buffering reagents for preparing buffers used in the analysis.

[0267] Certain other reagents and / or devices may be used for qualitative target protein analysis using an adelman degradation sequencer. Such reagents include phenyl isothiocyanate. Other reagents used in adelman degradation sequencing are well known in the art, and kits containing such reagents are within the scope of this disclosure.

[0268] In some cases, the kits disclosed herein are designed for the quantitative detection of one or more proteins in a sample. Some non-limiting examples include kits for the quantitative detection of one or more target proteins by immunoassay, mass spectrometry, and protein detection array analysis.

[0269] As described above, non-limiting examples of immunoassays include Western blotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), lateral flow immunochromatography, particle-based immunoassays, quantum dot-based immunoassays, etc. Therefore, certain embodiments of this disclosure provide kits for performing one or more of these assays on one or more target proteins disclosed herein.

[0270] In one embodiment, the kit includes a lateral flow immunochromatography device. Such a device allows for the detection and / or quantification of one or more target proteins: DLL1, VNN2, VAV3, and SUMF1. The lateral flow immunochromatography device may also include an internal control displaying a reference value or threshold.

[0271] In some cases, lateral flow immunochromatography devices may only allow the detection and / or quantification of one, two, three, or four target proteins from the following: DLL1, VNN2, VAV3, and SUMF1. In addition to the aforementioned one or more target proteins, lateral flow immunochromatography devices may allow the detection and / or quantification of one or more control proteins.

[0272] Some kits allow the detection of one or more target proteins using a protein detection array. Some of these protein detection arrays may include specific binding agents, such as antibodies that specifically bind to one or more of the following proteins: DLL1, VNN2, VAV3, and SUMF1. The antibodies are located at specific sites on a solid support. The kit may further include reagents that facilitate visualization of the specific binding between the target protein in the sample and the binding agent on the array. Such reagents include labeled secondary antibodies, such as those labeled with fluorescent dyes.

[0273] In some implementations, the kit allows for the detection of one or more target proteins via lysate microarray analysis. Such kits allow for the preparation of sample lysates and their immobilization on a support, such as a nitrocellulose-coated slide. The target protein is then detected using a labeled solution-phase-specific binder (e.g., a fluorescently labeled antibody that specifically binds to one or more target proteins). Therefore, the kit includes a labeled binder for one or more target proteins, particularly a fluorescently labeled antibody that specifically binds to one or more target proteins. In some cases, antibodies against different target proteins are differentially labeled, thereby allowing for multiplex detection and / or quantification of two or more target proteins.

[0274] In some cases, kits are designed to detect one of the following proteins in a sample: DLL1, VNN2, VAV3, and SUMF1. For example, some kits allow the detection of DLL1 in a sample. Some other kits allow the detection of VNN2 in a sample. Other implementations describe kits that allow the detection of VAV3 in a sample. There are even kits that allow the detection of SUMF1 in a sample.

[0275] In some cases, the kit is designed to detect any two of the following proteins in a sample: DLL1, VNN2, VAV3, and SUMF1, such as the following combinations: DLL1 and VNN2; DLL1 and VAV3; DLL1 and SUMF1; VNN2 and VAV3; VNN2 and SUMF1; and VAV3 and SUMF1.

[0276] In other embodiments, the kit is designed to detect any three of the following proteins in a sample: DLL1, VNN2, VAV3, and SUMF1, for example, the following combinations: DLL1, VNN2, and VAV3; DLL1, VAV3, and SUMF1; DLL1, VNN2, and SUMF1; and VNN2, VAV3, and SUMF1.

[0277] In some implementations, the kit is designed to detect all four proteins in a sample: DLL1, VNN2, VAV3, and SUMF1.

[0278] In some cases, the kit is designed to detect only one protein among DLL1, VNN2, VAV3, and SUMF1 in a sample, without detecting other proteins except for certain control proteins.

[0279] In some cases, the kit is designed to detect only two proteins in a sample of DLL1, VNN2, VAV3, and SUMF1, without detecting other proteins besides certain control proteins. For example, only the following protein combinations may be detected: DLL1 and VNN2; DLL1 and VAV3; DLL1 and SUMF1; VNN2 and VAV3; VNN2 and SUMF1; and VAV3 and SUMF1. In addition, one or more control proteins may be detected.

[0280] In other embodiments, the kit is designed to detect only three proteins of DLL1, VNN2, VAV3, and SUMF1 in a sample, without detecting other proteins except for certain control proteins. For example, the following protein combinations may be detected: DLL1, VNN2, and VAV3; DLL1, VAV3, and SUMF1; DLL1, VNN2, and SUMF1; and VNN2, VAV3, and SUMF1. In addition, one or more control proteins may be detected.

[0281] In some implementations, the kit is designed to detect only four proteins in a sample: DLL1, VNN2, VAV3, and SUMF1, without detecting other proteins except for certain control proteins.

[0282] In some cases, each specific binding member in the kits described herein independently comprises an antibody or its antigen-binding fragment, aptamer, or peptide-binding member. The one or more specific binding members are labeled with a detectable portion, such as an optically detectable portion.

[0283] In some cases, the kits disclosed herein are used in the methods disclosed herein, namely, to identify whether a subject is likely to respond positively to plasma exchange therapy for treating cognitive impairment in that subject.

[0284] In addition to the components described above, the kit of the present invention may also include instructions for carrying out the method of the present invention. These instructions may exist in the kit of the present invention in various forms, possibly one or more. One possible form of these instructions is as printed information on a suitable medium or substrate, such as printed paper, kit packaging, packaging inserts, etc. Another form is on a computer-readable medium, such as a disk, CD, portable flash drive, etc., on which the information is recorded. Another possible form is a website address, providing information about a remote site accessible via the Internet. In some cases, the kit may provide information about a smartphone application, which will subsequently provide relevant information. Any other convenient means of communication may also exist in the kit.

[0285] VI. Experimental Examples

[0286] A. Example 1 – Identification of proteins predicting responsiveness to plasma exchange therapy for the treatment of cognitive impairment

[0287] Example 1 describes an exemplary method for identifying protein biomarkers to predict a subject's response to plasma exchange (PE) therapy for the treatment of cognitive impairment. First, treatment benefit is confirmed by comparing changes in clinical outcomes between the treatment and placebo groups based on appropriate statistical tests (such as t-tests, ANOVA, MMRM, linear mixed models, chi-square tests, Fisher's exact test, logistic regression, etc.). Such analyses are also performed in subgroups if the sample size is sufficient.

[0288] In this embodiment, the Clinical Dementia Rating Scale Total Box Score (CDR.sb) was used as the clinical outcome. It is a holistic assessment of AD patients and a secondary endpoint of the AMBAR clinical trial. Six categories were assessed: memory, orientation, judgment and problem-solving, community affairs, home and hobbies, and personal care. A higher CDR.sb indicates more severe dementia.

[0289] A significant improvement in EOS (Elastic Occult Scale) was observed in the treatment group after PE treatment, with a decrease of 1.1 points (p = 0.002). A similar improvement (a decrease of 1.5 points) was observed in patients with moderate AD (p = 0.01), where both the PE and control groups experienced worsening, but PE treatment mitigated this worsening. In mild AD, the PE treatment group showed improvement, while the placebo group showed cognitive decline. (Boada et al., (2020), Alzheimer's & Dementia, 16:10, pp. 1412-1425.)

[0290] In a subset of patients, baseline predictive biomarkers for identifying responsiveness to clinical improvement in PE-Alb treatment were identified based on CDR.sb. Spearman correlation analysis was used to identify baseline plasma proteins associated with changes in CDR.sb at EOS. Such analyses demonstrate the strength of proteomic biomarkers associated with continuous clinical changes.

[0291] The Spearman correlation coefficient, or Spearman's rho (ρ), and its associated p-value are reported. A positive ρ indicates a positive correlation, and a negative ρ indicates a negative correlation.

[0292] An absolute value of ρ (|ρ|) > 0.3 is considered a true correlation, ρ between 0.3 and 0.5 is considered a low correlation, ρ between 0.5 and 0.7 is considered a moderate correlation, ρ between 0.7 and 0.9 is considered a high correlation, and ρ > 0.9 is considered a very high correlation. (See Mukaka (2012), Malawi Med J., 24(3): 69–71.)

[0293] Based on the concept of diagnostic testing, the CDR.sb changes over time (EOS) are transformed into two binary variables and compared to baseline: better or worse than baseline, and no worse than a clinically significant change (CDR.sb increases by 1 point). ROC is generated and AUC is calculated, and the diagnostic / predictive capabilities of candidate proteomic biomarkers are compared.

[0294] The optimal decision threshold (reference value) for high or low baseline protein levels was then determined based on the Youden index. Binary biomarkers (above or below the reference value) were then assessed based on their predictive power (measured by accuracy, sensitivity, and specificity). The estimated mean change in CDR.sb in each new group (high biomarker group and low biomarker group) was then calculated based on a linear mixture model (adjusted for age, sex, and baseline CDR.sb score).

[0295] These identified candidate biomarkers were validated through computer simulations, including testing in other clinical outcomes (ADAS_Cog and ADCS_ADL in the AMBAR study) and in randomly resampled samples (1001 resampled datasets (80% of the original sample size)).

[0296] Based on the above results, according to Figure 2B The flowchart shown prepares the final list of candidate biomarkers.

[0297] Selected single plasma protein biomarkers are provided in Table 4, with additional predictive information provided in Tables 5 and 6.

[0298] Table 4. Candidate plasma biomarkers identified based on Spearman correlation:

[0299]

[0300] Table 5. Predictive power of treatment benefit (CDR.Sb was better than baseline (>0) at EOS).

[0301]

[0302] Table 6. Predictive power of treatment benefit (CDR. Sb change <1 point relative to baseline, 2019 Diseaseseverity and minimal clinically important differences in clinical outcome assessments for Alzheimer's disease clinical trials). CMC: Clinically significant change.

[0303]

[0304] The data in Tables 5 and 6 indicate that DLL1 levels can be used as an exclusion criterion if the outcome after seeking treatment is better than baseline, due to its 92.5% specificity. If the deterioration after seeking treatment is less than 1 point, DLL1 levels can be used as a predictive biomarker, as it has a balanced sensitivity and specificity and an accuracy of 78.4%.

[0305] Table 7 shows the computer simulation validation in random sampling and other clinical outcomes for predicting AUCs that are better than baseline or at least worse than clinically meaningful changes (CMC).

[0306]

[0307] 1. What is the clinically relevant change on the ADAS-Cog? (2012) J. Neurosurg Psychiatry;83(2):171-3.

[0308] Figures 3A to 3C An example of predicting clinical changes from stratified objects derived from the candidate predictive biomarker baseline DLL1 is described.

[0309] Figures 4A to 4C An example of predicting clinical changes from a hierarchical object derived from a candidate predictive biomarker baseline VNN2 is described.

[0310] Figures 5A to 5C Examples of stratified subjects predicting clinical changes from baseline VAV3, a candidate predictive biomarker, are described.

[0311] Figures 6A to 6C Examples of stratified subjects predicting clinical changes from the candidate predictive biomarker baseline SUMF1 are described.

[0312] Despite the accompanying claims, this disclosure is also defined by the following terms:

[0313] 1. A method for analyzing samples obtained from an object, the method comprising:

[0314] The sample was tested for one or more proteins selected from the following: delta-like typical Notch ligand 1 (DLL1), vascular non-inflammatory molecule 2 (VNN2), guanine nucleotide exchange factor (VAV3), and sulfatase modifying factor 1 (SUMF1);

[0315] The subject in question has or is suspected of having cognitive impairment.

[0316] 2. The method according to Clause 1, which includes detecting DLL1 in the sample.

[0317] 3. The method according to Clause 1, which includes detecting VNN2 in the sample.

[0318] 4. The method according to Clause 1, which includes detecting VAV3 in the sample.

[0319] 5. The method according to Clause 1, which includes detecting SUMF1 in the sample.

[0320] 6. The method according to any one of Clauses 1 to 5, comprising qualitative detection of one or more proteins in the sample.

[0321] 7. The method according to any one of Clauses 1 to 5, comprising quantitative detection of one or more proteins in the sample.

[0322] 8. The method according to any one of Clauses 1 to 7, wherein the subject suffers from the aforementioned cognitive impairment.

[0323] 9. The method according to any one of Clauses 1 to 7, wherein the subject is suspected of having the cognitive impairment.

[0324] 10. The method according to any one of the preceding clauses, wherein the cognitive impairment is caused by a neurodegenerative disease.

[0325] 11. The method described in Clause 10, wherein the neurodegenerative disease is Alzheimer's disease (AD), Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, or vascular dementia.

[0326] 12. The method according to Clause 11, wherein the neurodegenerative disease is AD.

[0327] 13. The method according to any one of the preceding clauses, wherein the object is a candidate for plasma exchange therapy for treating the cognitive impairment.

[0328] 14. The method according to any one of the preceding clauses, comprising detecting one or more of the proteins in an immunoassay, mass spectrometry, or protein detection array analysis.

[0329] 15. The method according to any one of Clauses 1 to 13, comprising detecting one or more of the proteins in an aptamer-based multiplex proteomics assay.

[0330] 16. The method according to any one of the preceding clauses includes comparing the detection result with a threshold or reference value.

[0331] 17. The method according to any one of the preceding clauses, wherein the sample is a blood sample, serum sample, plasma sample or cerebrospinal fluid sample.

[0332] 18. The method according to any one of the preceding clauses, further comprising:

[0333] Based on protein expression data, it was determined whether the subject was likely to respond positively to plasma exchange therapy used to treat the subject's cognitive impairment.

[0334] 19. The method according to Clause 18, further comprising treating the cognitive impairment of the subject by administering the plasma exchange therapy to the subject if it is determined that the subject may respond positively to the plasma exchange therapy.

[0335] 20. A method for treating cognitive impairment in a subject, the method comprising:

[0336] Plasma exchange therapy is administered to the subject, wherein the subject is identified as likely to respond positively to plasma exchange therapy for the treatment of the cognitive impairment.

[0337] 21. The method according to Clause 20, wherein the subject is identified as potentially responding positively to plasma exchange therapy for the treatment of the cognitive impairment based on the detection results of one or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in the subject sample.

[0338] 22. The method according to Clause 20 or 21, wherein the subject is identified as potentially responding positively to plasma exchange therapy for the treatment of the cognitive impairment based on the detection results of two or more proteins selected from DLL1, VNN2, VAV3 and SUMF1 in the subject sample.

[0339] 23. The method according to any one of Clauses 20 to 22, wherein the subject is identified as potentially responding positively to plasma exchange therapy for the treatment of said cognitive impairment based on the detection results of three or more proteins selected from DLL1, VNN2, VAV3 and SUMF1 in the subject sample.

[0340] 24. The method according to any one of Clauses 20 to 23, wherein the subject is identified as potentially responding positively to plasma exchange therapy for the treatment of said cognitive impairment based on the detection results of all proteins of DLL1, VNN2, VAV3 and SUMF1 in the subject sample.

[0341] 25. The method according to Clause 20 or 24, wherein the object is identified as potentially responding positively to plasma exchange therapy for treating the cognitive impairment based on the detection results of DLL1 in the object sample.

[0342] 26. The method according to Clause 20 or 21, wherein the object is identified as potentially responding positively to plasma exchange therapy for treating the cognitive impairment based on the detection results of VNN2 in the object sample.

[0343] 27. The method according to Clause 20 or 21, wherein the subject is identified as potentially responding positively to plasma exchange therapy for the treatment of the cognitive impairment based on the detection results of VAV3 in the subject sample.

[0344] 28. The method according to Clause 20 or 21, wherein the subject is identified as potentially responding positively to plasma exchange therapy for the treatment of said cognitive impairment based on the detection results of SUMF1 in the subject sample.

[0345] 29. The method according to any one of Clauses 20 to 28, wherein the object is identified as potentially responding positively to plasma exchange therapy for treating the cognitive impairment based on an increase in the level of one or more proteins detected in a sample obtained from the object relative to a threshold, a reference value, or a reference sample.

[0346] 30. The method according to any one of Clauses 20 to 29, wherein the plasma exchange therapy comprises a total plasma exchange procedure.

[0347] 31. The method according to Clause 30, wherein the total plasma exchange comprises replacing substantially all of the subject's plasma with a first albumin solution.

[0348] 32. The method according to Clause 30 or 31, wherein the total plasma exchange procedure comprises weekly total plasma exchange for 5 to 8 weeks.

[0349] 33. The method according to any one of Clauses 30 to 32, wherein the total plasma exchange treatment comprises weekly total plasma exchange for 6 weeks.

[0350] 34. The method according to any one of clauses 31 to 33, wherein the first albumin solution contains 5% albumin.

[0351] 35. The method according to any one of clauses 30 to 34, wherein the plasma exchange therapy comprises performing a low-volume plasma exchange procedure using a second albumin solution after the total plasma exchange procedure.

[0352] 36. The method according to Clause 35, wherein the low-volume plasma exchange comprises replacing 20% ​​to 40% of the subject's plasma with the second albumin solution.

[0353] 37. The method according to Clause 35 or 36, wherein the low-volume plasma exchange procedure comprises monthly low-volume plasma exchange for at least 10 months.

[0354] 38. The method according to any one of clauses 35 to 37, wherein the low-volume plasma exchange procedure comprises monthly low-volume plasma exchange for 12 to 16 months.

[0355] 39. The method according to any one of clauses 35 to 38, wherein the second albumin solution comprises 20% albumin.

[0356] 40. A kit comprising reagents for measuring the amount of one or more proteins selected from DLL1, VNN2, VAV3 and SUMF1 in a sample.

[0357] 41. The kit according to Clause 40, comprising reagents for measuring the amount of DLL1 in the sample.

[0358] 42. The kit according to Clause 40, comprising reagents for measuring the amount of VNN2 in the sample.

[0359] 43. The kit according to Clause 40, comprising reagents for measuring the amount of VAV3 in the sample.

[0360] 44. The kit according to Clause 40, comprising reagents for measuring the amount of SUMF1 in the sample.

[0361] 45. A kit according to any one of clauses 40 to 44, wherein the reagent comprises one or more specific binding members that specifically bind to one or more corresponding proteins.

[0362] 46. ​​The kit according to Clause 45, wherein the one or more specific binding members are each independently an antibody or its antigen-binding fragment, aptamer or peptide binding member.

[0363] 47. The kit according to clause 45 or 46, wherein one or more specific binding members are labeled with a detectable portion.

[0364] 48. The kit according to Clause 47, wherein the detectable portion is an optically detectable portion.

[0365] 49. The kit according to any one of clauses 40 to 48, wherein the kit is suitable for performing immunoassays, Western blot analysis, mass spectrometry analysis or protein detection array analysis.

[0366] 50. The kit according to any one of clauses 40 to 48, wherein the kit is suitable for performing aptamer-based multiplex proteomics assays.

[0367] 51. The kit according to any one of clauses 40 to 50, wherein the kit is used to identify whether a subject is likely to respond positively to plasma exchange therapy for treating cognitive impairment in the subject.

[0368] In at least some of the previously described embodiments, one or more elements used in the embodiments may be used interchangeably in another embodiment, unless such substitution is technically impractical. Those skilled in the art will understand that various omissions, additions, and modifications can be made to the methods and structures described above without departing from the scope of the claimed subject matter. All such modifications and variations are intended to fall within the scope of the subject matter defined by the appended claims.

[0369] Those skilled in the art will understand that, in general, the terms used herein, particularly those in the appended claims (e.g., the body of the appended claims), are generally intended to be “open-ended” terms (e.g., the term “comprising” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “including” should be interpreted as “including but not limited to,” etc.). Those skilled in the art will further understand that if there is an intent to claim protection for a specific number of the referenced claim elements, that intent will be explicitly stated in the claims, and without such a statement, such intent does not exist. For example, to aid understanding, the appended claims below may contain the use of the introductory phrases “at least one” and “one or more” to introduce claim elements. However, the use of such phrases should not be construed as implying that introducing a claim element with the indefinite article “one” limits any particular claim containing such an introduced claim element to embodiments containing only one such element, even if the same claim includes the introductory phrases “one or more” or “at least one” and indefinite articles such as “one” (e.g., “one” should be interpreted as meaning “at least one” or “one or more”); the same applies to the use of definite articles to introduce claim elements. Furthermore, even when a specific number of elements of the introduced claims is explicitly referenced, those skilled in the art will recognize that such references should be interpreted as indicating at least the number referenced (e.g., simply referring to "two elements" without any other modifiers indicates at least two elements, or two or more elements). Additionally, when using conventions such as "at least one of A, B, and C," this structure is generally intended to have the meaning understood by those skilled in the art (e.g., "a system having at least one of A, B, and C" should include, but is not limited to, systems having a single A, a single B, a single C, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). When using conventions such as "at least one of A, B, or C," this structure is generally intended to have the meaning understood by those skilled in the art (e.g., "a system having at least one of A, B, or C" should include, but is not limited to, systems having a single A, a single B, a single C, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). Those skilled in the art will further understand that any extractive terms and / or phrases presenting two or more alternative terms, whether in the specification, claims, or drawings, should be understood to include the possibility of including one, any, or both terms. For example, the phrase "A or B" should be understood to include the possibility of including "A", "B", or "A and B".

[0370] Furthermore, when the features or aspects of this disclosure are described in accordance with the Markush Group, those skilled in the art will recognize that this disclosure is also described in accordance with any individual member or subgroup of members of the Markush Group.

[0371] As those skilled in the art will understand, for any and all purposes, such as providing a written description, all scopes disclosed herein also encompass any and all possible subscopes and combinations thereof. Any listed scope can be readily identified as adequately describing and inspiring the decomposition into at least two equal halves, thirds, quarters, quintiles, decimals, etc. As a non-limiting embodiment, each scope discussed herein can be readily decomposed into a lower third, a middle third, and an upper third, etc. As those skilled in the art will also understand, all language such as “up to,” “at least,” “greater than,” “less than,” etc., includes the referenced numbers and refers to a scope that can subsequently be decomposed into subscopes as described above. Finally, as those skilled in the art will understand, a scope includes each individual member. Thus, for example, a group having 1 to 3 items refers to a group having 1, 2, or 3 items. Similarly, a group having 1 to 5 items refers to a group having 1, 2, 3, 4, or 5 items, and so on.

[0372] Although the foregoing invention has been described in detail for clarity of understanding through illustration and embodiments, it will be apparent to those skilled in the art, based on the teachings of the present invention, that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

[0373] Therefore, the foregoing only illustrates the principles of the invention. It should be understood that those skilled in the art will be able to devise various arrangements that, while not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all embodiments and conditional language cited herein are primarily intended to aid the reader in understanding the principles of the invention and the conceptual contributions made by the inventors to advance the field, and should be interpreted as not being limited to these specifically cited embodiments and conditions. Moreover, all statements herein regarding the principles, aspects, and embodiments of the invention, as well as specific examples thereof, are intended to cover their structural and functional equivalents. Furthermore, such equivalents are contemplated to include both currently known equivalents and those developed in the future, i.e., any element performing the same function, regardless of its structure. Furthermore, regardless of whether such disclosure is expressly cited in the claims, nothing disclosed herein is intended to be offered to the public.

[0374] Therefore, the scope of the invention is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of the invention are embodied in the appended claims. In the claims, 35 USC §112(f) or 35 U.SC §112(6) is explicitly defined as being invoked for that definition only when the exact phrase “means for…” or the exact phrase “steps for…” is used in the definition of the claim; if such an exact phrase is not used in the definition of the claim, then 35 USC §112(f) or 35 USC §112(6) is not invoked.

Claims

1. A method for analyzing samples obtained from an object, the method comprising: The sample was tested for one or more proteins selected from the following: delta-like typical Notch ligand 1 (DLL1), vascular non-inflammatory molecule 2 (VNN2), guanine nucleotide exchange factor (VAV3), and sulfatase modifying factor 1 (SUMF1); The subject in question suffers from or is suspected of suffering from cognitive impairment.

2. The method of claim 2, further comprising qualitatively detecting one or more proteins in the sample.

3. The method according to claim 1 or 2, comprising quantitatively detecting one or more proteins in the sample.

4. The method according to any one of claims 1 to 3, wherein the subject suffers from or is suspected of suffering from the cognitive impairment.

5. The method according to any one of the preceding claims, wherein the cognitive impairment is caused by a neurodegenerative disease.

6. The method according to any one of the preceding claims, wherein the object is a candidate for plasma exchange therapy for treating the cognitive impairment.

7. The method according to any one of the preceding claims, comprising detecting one or more proteins in an immunoassay, mass spectrometry, protein detection array analysis, or aptamer-based multiplex proteomics assay.

8. The method according to any one of the preceding claims, comprising comparing the detection result with a threshold or a reference value.

9. The method according to any one of the preceding claims, wherein the sample is a blood sample, serum sample, plasma sample, or cerebrospinal fluid sample.

10. The method according to any one of the preceding claims, further comprising: Based on protein expression data, it was determined whether the subject was likely to respond positively to plasma exchange therapy used to treat the subject's cognitive impairment.

11. The method of claim 10, further comprising treating the cognitive impairment of the subject by administering the plasma exchange therapy to the subject if it is determined that the subject may respond positively to the plasma exchange therapy.

12. A method for treating cognitive impairment in a subject, the method comprising: Plasma exchange therapy is administered to the subject, wherein the subject is identified as likely to respond positively to plasma exchange therapy for the treatment of the cognitive impairment.

13. The method of claim 12, wherein the object is identified as potentially responding positively to plasma exchange therapy for treating the cognitive impairment based on the detection results of one or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in a sample of the object.

14. The method of claim 12 or 13, wherein the object is identified as potentially responding positively to plasma exchange therapy for the treatment of the cognitive impairment based on the detection results of two or more proteins selected from DLL1, VNN2, VAV3, and SUMF1 in a sample of the object.

15. A kit comprising reagents for measuring the amount of one or more proteins selected from DLL1, VNN2, VAV3 and SUMF1 in a sample.

Images