Blood tests to screen for Parkinson's disease

Abstract

In one embodiment, the disclosure relates to a method for excluding subjects from the need for diagnostic testing for Parkinson's disease (PD). In another embodiment, the disclosure relates to a method for excluding subjects from recruitment into a clinical study for an investigational PD drug. In yet another embodiment, the disclosure relates to a method for screening subjects to determine whether they are excluded from having PD, where subjects who cannot be excluded are administered a diagnostic test for PD, a treatment for PD, or a combination thereof.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 63 / 185,563, filed May 7, 2021, the disclosure of which is incorporated herein by reference in its entirety.

[0002] Statement of Federally Sponsored Research This invention was made with government support under grants AG054073 and AG058537 awarded by the National Institutes of Health. The government has certain rights in this invention. [Background technology]

[0003] Parkinson's disease (PD) is the second most common neurodegenerative disorder in the United States (US), affecting more than 1% of individuals aged 65 years or older. The cost of PD to society was reported to be $23 billion annually in the US in 2005. Given the rapid growth of the aging population, these costs will continue to increase over the coming decades. The most accurate diagnosis of PD is achieved through specialty clinics, where clinical evaluation and advanced neurodiagnostic procedures are costly, time-consuming, and invasive. In the US, primary care clinics serve as "gatekeepers" to specialty clinics, and these frontline primary care practitioners provide referrals for advanced diagnostic procedures. However, the average primary care consultation time is approximately 18 minutes, making a detailed neurological examination difficult.

[0004] In 2017, community-dwelling PD patients and general practitioners (GPs) were interviewed to understand their views on the role of primary care in managing PD. Disagreements between patients and GPs were found, with patients feeling that GPs lacked skilled knowledge or skills and undermined the GP's role in patients with advanced PD stages. On the other hand, GPs respected patient autonomy in making early-stage decisions but felt that a more active role for GPs was justified in more advanced PD stages. One conclusion of the study was that patients would likely benefit from a more holistic approach provided by GPs when combined with specialized care. However, there are currently no rapid, cost-effective screening tools for primary care providers to use in their daily activities to rule out PD among their patients and thus prevent further diagnostic testing for PD. Summary of the Invention [Problem to be solved by the invention]

[0005] Thus, there is a need in the art for faster, cost-effective methods for screening subjects to rule out PD and to prevent them from undergoing more invasive and / or expensive testing for Parkinson's disease. The present disclosure fulfills this unmet need. [Means for solving the problem]

[0006] In one aspect, the disclosure provides a method for eliminating a subject from the need for diagnostic testing for Parkinson's disease, comprising the steps of: (a) obtaining a blood, plasma, or serum sample from the subject; (b) detecting in the blood, plasma, or serum sample any of interleukin (IL)-7, tumor necrosis factor alpha (TNFα), IL-5, IL-6, C-reactive protein (CRP), IL-10, soluble intracellular adhesion molecule (sICAM-1), Factor VII, I309, alpha-2-microglobulin (A2M), chemokine (CC motif) ligand 17 (TARC), eotaxin 3, soluble vascular cell adhesion molecule 1 (sVCAM-1), factor VII, I309, alpha-2-microglobulin (A2M), factor VII, factor VII, factor VIII ... (c) measuring the expression levels of one or more biomarkers selected from the group consisting of serum amyloid A1 cluster (SAA), thrombopoietin (TPO), fatty acid binding protein (FABP), IL-18, beta-2-microglobulin (B2M), serum amyloid A1 cluster (SAA), tenacin C, tumor necrosis factor receptor 1 (TNFR1), and any pancreatic polypeptide (PPY); (c) comparing the expression levels of the one or more biomarkers in (b) with a representative statistical sample of subjects, thereby determining whether the subject can be excluded from diagnostic testing for Parkinson's disease; and (d) determining that the subject should be excluded from diagnostic testing for Parkinson's disease based on the comparing step.In certain embodiments, the method further comprises (e) avoiding, not initiating, or discontinuing diagnostic testing for Parkinson's disease, wherein the diagnostic testing is selected from the group consisting of a neurological exam, an MRI, a dopamine transporter (DAT) scan, a brain ultrasound, a PET scan, a detailed neuropsychological testing, and any combination thereof. In certain embodiments, the method further comprises (f) avoiding, not initiating, or discontinuing treatment for Parkinson's disease, wherein the treatment is selected from the group consisting of levodopa, a dopamine agonist, a glutamate agonist, an anticholinergic, a catechol-o-methyl transferase (COMT) inhibitor, a monoamine oxidase type B (MAO-B) inhibitor, a dopamine therapy, an N-methyl-D-aspartate (NMDA) antagonist, and any combination thereof. In certain embodiments, the one or more biomarkers are selected from the group consisting of tenascin-C, IL-6, I309, IL-7, and FABP. In certain embodiments, the expression level of each biomarker in the group consisting of tenascin-C, IL-6, I309, and IL-7 is measured, and optionally the expression level of one or more biomarkers selected from the group consisting of FABP, TNFα, IL-5, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, IL-18, B2M, SAA, and PPY is measured. In certain embodiments, the expression level of the one or more biomarkers is measured using electrochemiluminescence.In certain embodiments, if the expression level of one or more biomarkers in (i)(b) is statistically similar to the mean expression level of the corresponding one or more biomarkers obtained from a group of individuals in the statistical sample who do not have Parkinson's disease, the subject is excluded from the diagnostic test for Parkinson's disease; or if the expression level of one or more biomarkers in (ii)(b) is not statistically similar to the mean expression level of the corresponding one or more biomarkers obtained from a group of individuals in the statistical sample who have been diagnosed with Parkinson's disease, the subject is excluded from the diagnostic test for Parkinson's disease.

[0007] In another aspect, the disclosure provides a method for excluding subjects from recruitment into a clinical trial for an investigational Parkinson's disease drug, comprising the steps of: (a) obtaining a blood, plasma, or serum sample from the subject; (b) detecting in the blood, plasma, or serum sample any of interleukin (IL)-7, tumor necrosis factor alpha (TNFα), IL-5, IL-6, C-reactive protein (CRP), IL-10, soluble intercellular adhesion molecule (sICAM-1), Factor VII, I309, alpha-2-microglobulin (A2M), chemokine (CC motif) ligand 17 (TARC), eotaxin 3, soluble vascular cell adhesion molecule 1 (sVCAM-1), thrombopoietin, ... (c) measuring the expression levels of one or more biomarkers selected from the group consisting of ATP, ATP-dependent ATPase inhibitor (TPO), fatty acid binding protein (FABP), IL-18, beta-2-microglobulin (B2M), serum amyloid Al cluster (SAA), tenascin-C, tumor necrosis factor receptor 1 (TNFR1), and any pancreatic polypeptide (PPY); (c) comparing the expression levels of the one or more biomarkers in (b) with a representative statistical sample of subjects; and (d) excluding a subject from recruitment into a clinical trial if the comparison with the statistical sample excludes the subject from having Parkinson's disease. In certain embodiments, the method further comprises (e) avoiding, not initiating, or discontinuing diagnostic testing for Parkinson's disease, wherein the diagnostic testing is selected from a neurological exam, an MRI, a dopamine transporter (DAT) scan, a brain ultrasound, a PET scan, and a detailed neuropsychological test. In certain embodiments, the method further comprises (f) avoiding, not initiating, or discontinuing treatment for Parkinson's disease, wherein the treatment is selected from levodopa, dopamine agonists, glutamate agonists, anticholinergics, catechol-o-methyltransferase (COMT) inhibitors, monoamine oxidase type B (MAO-B) inhibitors, dopamine therapy, N-methyl-D-aspartate (NMDA) antagonists, and any combination thereof.In certain embodiments, the one or more biomarkers are selected from the group consisting of tenascin-C, IL-6, I309, IL-7, and FABP. In certain embodiments, the expression level of each biomarker in the group consisting of tenascin-C, IL-6, I309, and IL-7 is measured, and optionally the expression level of one or more biomarkers selected from the group consisting of FABP, TNFα, IL-5, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, IL-18, B2M, SAA, and PPY is measured. In certain embodiments, the expression level of the one or more biomarkers is measured using electrochemiluminescence. In certain embodiments, if the expression level of the one or more biomarkers in (i)(b) is statistically similar to the mean expression level of the corresponding one or more biomarkers obtained from a group of individuals in the statistical sample who do not have Parkinson's disease, the subject is excluded from recruitment into the clinical study; or if the expression level of the one or more biomarkers in (ii)(b) is not statistically similar to the mean expression level of the corresponding one or more biomarkers obtained from a group of individuals in the statistical sample who have been diagnosed with Parkinson's disease, the subject is excluded from recruitment into the clinical study.

[0008] In another aspect, the disclosure provides a method for screening a subject to determine whether the subject is ruled out as having Parkinson's disease, comprising the steps of: (a) obtaining a blood, plasma, or serum sample from the subject; (b) detecting in the blood, plasma, or serum sample any of interleukin (IL)-7, tumor necrosis factor alpha (TNFα), IL-5, IL-6, C-reactive protein (CRP), IL-10, soluble intercellular adhesion molecule (sICAM-1), Factor VII, I309, alpha-2-microglobulin (A2M), chemokine (CC motif) ligand 17 (TARC), eotaxin 3, soluble vascular cell adhesion molecule 1 (sVCAM-1), thrombopoietin (TPO), fatty acid binding protein (FABP), IL-18, beta-2-microglobulin (B2M), serum amyloid A1, IL-20, IL-12, IL-14, IL-16, IL-18, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-24, IL-25, IL-26, IL-27, IL-28, IL-29, IL-30, IL-31, IL-32, IL-33, IL-34, IL-35, IL-36, IL-37, IL-38, IL-39, IL-40, IL-41, IL-42, IL-43, IL-44, IL-45, IL-46, IL-47, IL-48, IL-49, IL-50, IL-51, IL-52, IL-53, IL-54, IL-55, IL-56, IL-57, IL-58, (c) measuring the expression levels of one or more biomarkers selected from the group consisting of ATP-dependent ATPase inhibitors (ATPase inhibitors), ... In certain embodiments, the therapy is selected from the group consisting of levodopa, dopamine agonists, glutamate agonists, anticholinergics, catechol-o-methyltransferase (COMT) inhibitors, monoamine oxidase type B (MAO-B) inhibitors, dopamine therapy, N-methyl-D-aspartate (NMDA) antagonists, and any combination thereof.In certain embodiments, the one or more biomarkers are selected from the group consisting of tenascin-C, IL-6, I309, IL-7, and FABP. In certain embodiments, the expression level of each biomarker in the group consisting of tenascin-C, IL-6, I309, and IL-7 is measured, and optionally the expression level of one or more biomarkers selected from the group consisting of FABP, TNFα, IL-5, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, IL-18, B2M, SAA, and PPY is measured. In certain embodiments, the expression level of the one or more biomarkers is measured using electrochemiluminescence. In certain embodiments, if (i) the expression level of one or more biomarkers in (b) is not statistically similar to the mean expression level of the corresponding one or more biomarkers obtained from a group of individuals in the statistical sample who do not have Parkinson's disease, the subject is not ruled out as having Parkinson's disease; or (ii) if the expression level of one or more biomarkers in (b) is statistically similar to the mean expression level of the corresponding one or more biomarkers obtained from a group of individuals in the statistical sample who have been diagnosed with Parkinson's disease, the subject is not ruled out as having Parkinson's disease. In certain embodiments, the method further comprises the step of (f) referring a subject not ruled out as having Parkinson's disease to a specialist in Parkinson's disease.

[0009] In another aspect, the disclosure provides a method for eliminating a subject from the need for diagnostic testing for Parkinson's disease, comprising the steps of: (a) obtaining a blood, plasma, or serum sample from the subject; (b) detecting in the blood, plasma, or serum sample any of the following: interleukin (IL)-7, tumor necrosis factor alpha (TNFα), IL-5, IL-6, C-reactive protein (CRP), IL-10, soluble intercellular adhesion molecule (sICAM-1), Factor VII, I309, alpha-2-microglobulin (A2M), chemokine (CC motif) ligand 17 (TARC), eotaxin 3, soluble vascular cell adhesion molecule 1 (sVCAM-1), thrombopoietin (TPO), fatty acid binding protein (FABP), I The method includes the steps of: (a) measuring the expression level of one or more biomarkers selected from the group consisting of L-18, beta-2-microglobulin (B2M), serum amyloid Al cluster (SAA), tenascin-C, TNRF1, and any pancreatic polypeptide (PPY); (b) using a computer to compare the expression levels of the one or more biomarkers in (b) with a representative statistical sample of subjects to determine whether the subject can be excluded from having Parkinson's disease; and (c) using a computer to determine whether the subject should be excluded from diagnostic testing for Parkinson's disease based on the comparing step. In certain embodiments, the method further includes the step of (e) avoiding, not initiating, or discontinuing diagnostic testing for Parkinson's disease, wherein the diagnostic testing is selected from the group consisting of a neurological exam, an MRI, a dopamine transporter (DAT) scan, a brain ultrasound, a PET scan, a detailed neuropsychological test, and any combination thereof. In certain embodiments, the method further comprises (f) avoiding, not initiating, or discontinuing treatment for Parkinson's disease, wherein the treatment is selected from the group consisting of levodopa, dopamine agonists, glutamate agonists, anticholinergics, catechol-o-methyltransferase (COMT) inhibitors, monoamine oxidase type B (MAO-B) inhibitors, dopamine therapy, N-methyl-D-aspartate (NMDA) antagonists, and any combination thereof.In certain embodiments, the one or more biomarkers are selected from the group consisting of tenascin-C, IL-6, I309, IL-7, and FABP. In certain embodiments, the expression level of each biomarker in the group consisting of tenascin-C, IL-6, I309, and IL-7 is measured, and optionally the expression level of one or more biomarkers selected from the group consisting of FABP, TNFα, IL-5, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, IL-18, B2M, SAA, and PPY is measured. In certain embodiments, the expression level of the one or more biomarkers is measured using electrochemiluminescence. In certain embodiments, if the expression level of one or more biomarkers in (i)(b) is determined by the computer to be statistically similar to the mean expression level of the corresponding one or more biomarkers obtained from a group of individuals in the statistical sample who do not have Parkinson's disease, the subject is excluded from the diagnostic test for Parkinson's disease; or if the expression level of one or more biomarkers in (ii)(b) is determined by the computer to be not statistically similar to the mean expression level of the corresponding one or more biomarkers obtained from a group of individuals in the statistical sample who have been diagnosed with Parkinson's disease, the subject is excluded from the diagnostic test for Parkinson's disease. [Brief explanation of the drawings]

[0010] The following detailed description of selected embodiments of the invention will be better understood when read in conjunction with the accompanying drawings. For the purpose of illustrating the invention, there is shown in the drawings illustrative embodiments. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings. [Figure 1]FIG. 1 shows accuracy and variable importance plots for the Parkinson's disease blood test (PDBT) based on a support vector machine (SVM) of the training set. [Figure 2] FIG. 1 shows the accuracy and ROC curves of SVM-based PDBT on the test set. DETAILED DESCRIPTION OF THE INVENTION

[0011] While the making and use of various embodiments of the invention are discussed in detail below, it should be recognized that the present invention provides many applicable inventive concepts that can be embodied in a wide variety of specific contexts. The specific embodiments discussed herein are merely illustrative of specific ways to make and use the invention, and do not delimit the scope of the invention.

[0012] To facilitate understanding of the present invention, several terms are defined below. Terms defined herein have meanings commonly understood by one of ordinary skill in the art relevant to the present invention. Terms such as "a," "an," and "the" are not intended to refer to a singular entity only, but include the general class of which a specific example may be used for illustration. While terminology herein is used to describe particular embodiments of the present invention, their usage does not delimit the scope of the invention except as outlined in the claims.

[0013] The differential diagnosis of neurodegenerative diseases is challenging but crucial for clinical treatment and management, as well as for designing therapeutic and preventative trials. Appropriate referrals are usually required from primary care providers to refer patients to specialty clinics for diagnostic testing and treatment. However, previous studies have demonstrated that the evaluation and management of neurodegenerative diseases is inadequate in primary care settings, with inappropriate medications frequently administered. Given that the average physician consultation time in an outpatient setting for individuals over 65 years of age is approximately 18 minutes, primary care providers desperately need rapid, cost-effective screening to rule out neurological conditions in their patients, which can then result in appropriate referrals to specialists.

[0014] The availability of a blood-based screening tool that can be implemented within a primary care clinic setting would have a significant impact. From a clinical perspective, fewer than half of the physicians surveyed considered screening for neurodegenerative diseases important, while the majority of laypeople and caregivers considered such screening very important. In addition, the average physician consultation lasts less than 20 minutes for geriatric patients in outpatient settings, significantly limiting the time available for even brief neurological and cognitive assessments. Therefore, primary care providers desperately need screening tools to exclude patients from specialist referral for diagnostic testing for possible neurodegenerative diseases. Numerous studies have been conducted demonstrating the utility of diagnostic tests for neurodegenerative diseases (MRI, fMRI, DTI, PET), but they are cost-prohibitive and time-consuming to rule out disease in a primary care setting.

[0015] Within the primary care setting, the role of screening tests is to exclude patients who do not require additional medical procedures or diagnostic follow-up, thereby reducing stress and containing costs. The application of specialty clinic-based diagnostic tests as screens to rule out disease in primary care settings may seem straightforward, but it is not, and as demonstrated below, the previous procedures would not work within the primary care setting. The ability to perform blood-based screens to rule out disease in primary care settings is important, but is highly complicated due to the substantially lower base rate of disease presence compared to specialty clinics, which has a tremendous impact on the predictive accuracy of the test results.

[0016] Several recent advances have been made in the development of biomarker-based tests to detect individuals with neurodegenerative diseases and distinguish them from other neurodegenerative diseases, including Alzheimer's disease (AD), PD, dementia with Lewy bodies (DLB), and AD among adults with Down syndrome (DS-AD). Over the past few decades, the search for biomarkers with diagnostic and predictive utility in neurodegenerative diseases has increased exponentially, with the majority of studies focusing on neuroimaging and cerebrospinal (CSF) methodologies. Indeed, dopamine transporter single-photon emission CT (DaT-SPECT) has been approved as a test for diagnosing PD. In addition to blood-based biomarkers, research suggests that CSF markers may also hold utility in the differential diagnosis of neurodegenerative diseases. Although advanced neuroimaging, blood-based markers, and CSF methods have tremendous potential as diagnostic tests for PD, invasiveness, accessibility, cost barriers, and technical feasibility have so far limited the utility of such methods to diagnostic testing applications. To date, these methods have not been found suitable as screening methods to rule out disease in primary care settings.

[0017]

[0003] The present disclosure relates, in part, to methods for excluding a subject from the need for diagnostic testing for Parkinson's disease. In some embodiments, the present disclosure provides methods for excluding PD in a subject. In some embodiments, the disclosed methods can be used to rule out PD in a subject in a primary care setting. In some embodiments, the methods include measuring the expression level of one or more biomarkers selected from the group consisting of IL-7, TNFα, IL-5, IL-6, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, FABP3, IL18, B2M, SAA, tenascin-C, TNFR1, and optionally PPY.

[0018] In some embodiments, one or more biomarkers include IL-7. In some embodiments, one or more biomarkers include TNFα. In some embodiments, one or more biomarkers include IL-5. In some embodiments, one or more biomarkers include IL-6. In some embodiments, one or more biomarkers include CRP. In some embodiments, one or more biomarkers include IL-10. In some embodiments, one or more biomarkers include sICAM-1. In some embodiments, one or more biomarkers include Factor VII. In some embodiments, one or more biomarkers include I309. In some embodiments, one or more biomarkers include A2M. In some embodiments, one or more biomarkers include TARC. In some embodiments, one or more biomarkers include eotaxin 3. In some embodiments, one or more biomarkers include sVCAM-1. In some embodiments, one or more biomarkers include TPO. In some embodiments, one or more biomarkers include FABP3. In some embodiments, the one or more biomarkers include IL18. In some embodiments, the one or more biomarkers include B2M. In some embodiments, the one or more biomarkers include SAA. In some embodiments, the one or more biomarkers include tenascin-C. In some embodiments, the one or more biomarkers include TNFR1. In some embodiments, the one or more biomarkers include PPY.

[0019] In some embodiments, the one or more biomarkers include tenascin-C. In some embodiments, the one or more biomarkers include tenascin-C and IL-6. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, and I309. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, and IL-7. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, and FABP3. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, and TARC. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, and TNFα. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, and A2M. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, and TPO. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, and IL5. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, and Factor VII. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, and IL10.In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3, and SAA. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3, SAA, and IL 18. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3, SAA, IL18, and sICAM-1. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3, SAA, IL18, sICAM-1, and CRP.

[0020] In certain embodiments, the expression levels of selected biomarkers are measured in a blood, plasma, or serum sample obtained from the subject. In some embodiments, the expression levels of one or more biomarkers are compared to the expression levels of one or more corresponding biomarkers in a representative statistical sample of subjects, where the comparison is used to determine whether the subject warrants diagnostic screening for Parkinson's disease or whether the subject can be excluded from diagnostic testing for Parkinson's disease. In some embodiments, the method for excluding a subject from the need for diagnostic testing for Parkinson's disease can be performed in a primary care setting.

[0021] In another aspect, the disclosure relates to a method for excluding subjects from recruitment into a clinical trial of an investigational Parkinson's disease drug, hi some embodiments, the method comprises measuring the expression level of one or more biomarkers selected from the group consisting of IL-7, TNFα, IL-5, IL-6, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, FABP3, IL18, B2M, SAA, tenascin-C, TNRF1, and optionally PPY.

[0022] In some embodiments, one or more biomarkers include IL-7. In some embodiments, one or more biomarkers include TNFα. In some embodiments, one or more biomarkers include IL-5. In some embodiments, one or more biomarkers include IL-6. In some embodiments, one or more biomarkers include CRP. In some embodiments, one or more biomarkers include IL-10. In some embodiments, one or more biomarkers include sICAM-1. In some embodiments, one or more biomarkers include Factor VII. In some embodiments, one or more biomarkers include I309. In some embodiments, one or more biomarkers include A2M. In some embodiments, one or more biomarkers include TARC. In some embodiments, one or more biomarkers include eotaxin 3. In some embodiments, one or more biomarkers include sVCAM-1. In some embodiments, one or more biomarkers include TPO. In some embodiments, one or more biomarkers include FABP3. In some embodiments, the one or more biomarkers include IL18. In some embodiments, the one or more biomarkers include B2M. In some embodiments, the one or more biomarkers include SAA. In some embodiments, the one or more biomarkers include tenascin-C. In some embodiments, the one or more biomarkers include TNFR1. In some embodiments, the one or more biomarkers include PPY.

[0023] In some embodiments, the one or more biomarkers include tenascin-C. In some embodiments, the one or more biomarkers include tenascin-C and IL-6. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, and I309. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, and IL-7. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, and FABP3. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, and TARC. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, and TNFα. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, and A2M. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, and TPO. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, and IL5. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, and Factor VII. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, and IL10.In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3, and SAA. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3, SAA, and IL 18. In some embodiments, the one or more biomarkers comprise tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3, SAA, IL18, and sICAM-1. In some embodiments, the one or more biomarkers include tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL5, Factor VII, IL10, sVCAM-1, eotaxin 3, SAA, IL18, sICAM-1, and CRP.

[0024] In some embodiments, the expression levels of one or more biomarkers are compared to the expression levels of corresponding one or more biomarkers in a representative statistical sample of subjects, where the comparison is used to determine whether a subject can be ruled out as having Parkinson's disease and therefore should be excluded from the clinical trial.

[0025] definition 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 belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described.

[0026] As used herein, each of the following terms has the meaning associated with it in this section.

[0027] The articles "a" and "an" are used herein to refer to one or more than one (i.e., at least one) of the object of the article. By way of example, "an element" means one element or more than one element. As used herein, "about," when referring to a measurable value, e.g., amount, time, etc., is meant to encompass a variation of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and even more preferably ±0.1% from the specified value, where such variation is appropriate for performing the disclosed method.

[0028] As used herein, the phrases "primary care clinic," "primary care setting," and "primary care provider" are used interchangeably to refer to the primary point of patient contact / consultation within the health care system, connecting the patient with specialists they may need.

[0029] As used herein, the term "specialist" refers to a medical practice or physician that specializes in a particular disorder, such as a neurological disorder, a psychiatric disorder, or even more specifically, a movement disorder or memory disorder.

[0030] As used herein, the phrases "screen out" or "rule out" refer to a blood, serum, or plasma test that rules out Parkinson's disease (PD). Conversely, subjects who are not screened out or ruled out as having PD would be recommended for accurate diagnostic testing for PD. As such, the present disclosure is a screening test, not a diagnostic test. The present disclosure further provides an initial screening blood test that can be used in a primary care setting to screen out PD.

[0031] As used herein, the following abbreviations are used and can include mammalian versions of these genes or gene products, although in certain embodiments, the gene or gene product is a human gene or gene product: IL7 - interleukin-7, TNFα - tumor necrosis factor alpha, IL5 - interleukin-5, IL6 - interleukin-6, CRP - C-reactive protein, IL10 - interleukin-10, TNC - tenascin C, ICAM1 - intracellular adhesion molecule 1, FVII - factor VII, I309 - chemokine (CC motif) ligand 1, TNFR1 - tumor necrosis factor receptor 1, A2M - alpha-2-microglobulin, TARC - chemokine (CC motif) ligand 17, eotaxin 3, VCAM1 - vascular cell adhesion molecule 1, TPO - thrombopoietin, FABP3 - fatty acid binding protein 3, IL18 - interleukin-18, B2M - beta-2-microglobulin, SAA - serum amyloid A1 cluster, PPY - pancreatic polypeptide, DJ1 - Parkinson protein 7, α-syn - α-synuclein.

[0032] As used herein, FABP and FABP3 are interchangeable.

[0033] As used herein, intracellular adhesion molecule 1 (ICAM1, ICAM-1) is interchangeable with soluble intercellular adhesion molecule 1 (sICAM1, sICAM-1).

[0034] As used herein, vascular cell adhesion molecule 1 (VCAM1, VCAM-1) is interchangeable with soluble vascular cell adhesion molecule 1 (sVCAM1, sVCAM-1).

[0035] As used herein, the phrase "neurological disease" refers to a disease or disorder of the central nervous system, many of which can include neurodegenerative disorders such as Alzheimer's disease (AD), Parkinson's disease (PD), mild cognitive impairment (MCI), frontotemporal dementia (FTD), dementia with Lewy bodies (DLB), Down's syndrome (DS), and dementias and neurological diseases, including multiple sclerosis and neuropathies.

[0036] As used herein, the terms "Parkinson's disease patient" and "individual diagnosed with Parkinson's disease" all refer to an individual who has been diagnosed with PD or has received a diagnosis of Parkinson's disease.

[0037] As used herein, the phrase "Parkinson's disease biomarker" refers to a biomarker used to screen out / rule out Parkinson's disease (PD).

[0038] As used herein, the term "Parkinson's disease biomarker protein" refers to any protein biomarker or substance that is functional at the level of the protein biomarker that is measured to screen out / rule out Parkinson's disease (PD).

[0039] As used herein, a "blood sample" refers to a biological sample derived from blood, preferably peripheral blood (or circulating blood). A blood sample may be, for example, whole blood, serum, or plasma. In certain embodiments, serum is preferred as a source for biomarkers because the sample is readily available, often obtained for other sampling, stable, requires little processing, making it ideal for locations with little refrigeration or electricity, easily transportable, and commonly handled by medical support staff.

[0040] As used herein, a "normal" individual or a sample from a "normal" individual refers to quantitative data, qualitative data, or both, from an individual who has been or will be assessed by a physician to be free of a disease, e.g., a neurological disease. Often, the "normal" individual will also be age-matched within 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years to the sample from the individual being assessed.

[0041] As used herein, the term "treatment" refers to the alleviation, amelioration, and / or stabilization of symptoms of a particular disorder, as well as the delay in the progression of the symptoms. For example, "treatment" of PD includes any one or more of the following: (1) elimination of one or more symptoms of PD, (2) reduction of one or more symptoms of PD, (3) stabilization of symptoms of PD (e.g., not progressing to a more advanced stage of PD), and (4) delay in the onset of one or more symptoms of PD; and (5) delay in the progression (i.e., worsening) of one or more symptoms of PD.

[0042] As used herein, the phrase "neurocognitive assessment" is used to describe one or more tests known to those of skill in the art for measuring cognitive status or impairment, and may include, but is not limited to, the 4-point clock drawing test, verbal fluency test, trail making test, list learning test, etc. Those of skill in the art will recognize and know how these tests can be modified, and how new tests measuring similar cognitive function can be developed and administered for use in accordance with the present invention.

[0043] explanation method In one aspect, the present disclosure provides a method for excluding a subject from the need for a diagnostic test for Parkinson's disease. In certain embodiments, the method includes the steps of: (a) obtaining a blood, plasma, or serum sample from the subject; (b) measuring, in the blood, plasma, or serum sample, the expression level of one or more biomarkers selected from the group consisting of IL-7, TNFα, IL-5, IL-6, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, FABP3, IL18, B2M, SAA, tenascin-C, TNFR1, and optionally PPY; (c) comparing the expression level of the one or more biomarkers in (b) with a representative statistical sample of subjects, thereby determining whether the subject can be excluded from the diagnostic test for Parkinson's disease; and (d) determining that the comparing step indicates that the subject should be excluded from the diagnostic test for Parkinson's disease.

[0044] In some embodiments, the method further comprises avoiding, not initiating, or discontinuing diagnostic testing for Parkinson's disease, wherein the diagnostic testing is selected from MRI, dopamine transporter (DAT) scan, brain ultrasound, PET scan, and detailed neuropsychological testing. In some embodiments, the method further comprises avoiding, not initiating, or discontinuing treatment for Parkinson's disease, wherein the treatment is selected from levodopa, dopamine agonists, glutamate agonists, anticholinergics, catechol-o-methyltransferase (COMT) inhibitors, monoamine oxidase type B (MAO-B) inhibitors, dopamine therapy, N-methyl-D-aspartate (NMDA) antagonists, and any combination thereof.

[0045] In certain embodiments, the subject is an elderly subject. In other embodiments, the subject is a middle-aged subject. In some embodiments, the subject has concerns about changes in movement (self-reported, doctor-reported, or otherwise) or a family history of PD. In certain embodiments, the middle-aged subject is 30 years of age or older. In certain embodiments, the subject is a human subject.

[0046] In certain embodiments, the sample is a blood sample. In some embodiments, the blood, plasma, or serum sample is obtained from a subject in a primary care setting.

[0047] In some embodiments, the statistical sample comprises a group of individuals aged 30 years or older who do not have a neurological disease or disorder. In other embodiments, the statistical sample comprises a group of individuals aged 30 years or older who have been diagnosed with a neurological disease or disorder. In some embodiments, the statistical sample comprises a group of individuals aged 30 years or older who have been diagnosed with PD. In some embodiments, the statistical sample comprises a measurement of the expression level of one or more biomarkers selected from IL-7, TNFα, IL-5, IL-6, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, FABP3, IL18, B2M, SAA, tenascin-C, TNRF1, and optionally PPY from each individual in the statistical sample.

[0048] In certain embodiments, a statistically similar expression level of one or more biomarkers in a subject's sample compared to corresponding biomarkers from individuals in the statistical sample who do not have a neurological disease or disorder indicates that diagnostic testing of the subject for PD is not necessary. In another embodiment, a statistically similar expression level of one or more biomarkers in a subject's sample compared to corresponding biomarkers from individuals in the statistical sample who are diagnosed with PD indicates that diagnostic testing of the subject for PD is necessary. In some embodiments, the expression level of one or more biomarkers measured in the subject's sample is compared to the mean expression level of the corresponding biomarkers in the statistical sample for each group of individuals in the statistical sample.

[0049] In certain embodiments, where the expression level of one or more measured biomarkers indicates that the subject needs diagnostic testing for PD, the subject is referred to a PD specialist for diagnostic testing. In other embodiments, where the expression level of one or more measured biomarkers indicates that the subject needs diagnostic testing, the subject is not referred to a PD specialist for diagnostic testing. Thus, in some embodiments, the disclosed methods prevent the subject from undergoing invasive and expensive testing that a specialist could prescribe to determine whether the subject has PD.

[0050] In some embodiments, the step of measuring the expression level of one or more biomarkers in the blood, plasma, or serum sample comprises measuring the expression level of one or more biomarkers of tenascin-C; tenascin-C and IL-6; tenascin-C, IL-6, and I309; tenascin-C, IL-6, I309, and IL-7; tenascin-C, IL-6, I309, IL-7, and FABP; tenascin-C, IL-6, I309, IL-7, FABP3, and TARC; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, and TNFα; tenascin-C, IL- tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, and TPO; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, and A2M; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, and IL-5; Tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, and factor VII; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, and IL-10; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, and sVCAM-1; tenascin-C, IL-6, I309, IL-7, FABP3 P3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, eotaxin 3, and SAA; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3;Tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, and IL-18; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3; tenascin-C, IL-6, I309, IL-7, FABP In some embodiments, the method comprises measuring the expression levels of the following biomarkers: IL-3, TARC, TNFα, A2M, TPO, B2M, IL-5, Factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, IL-18, and sICAM-1; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, Factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, IL-18, sICAM-1, and CRP. In some embodiments, the expression levels of each of the measured biomarkers are compared with the expression levels of the corresponding biomarkers in a statistical sample to determine whether the subject should be excluded from further testing for Parkinson's disease.

[0051] In certain embodiments, the expression levels of one or more of tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M are measured in a blood, plasma, or serum sample. In certain embodiments, the expression levels of each of tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M are measured in a blood, plasma, or serum sample. In certain embodiments, the expression levels of each of tenascin-C, IL-6, I309, and IL-7, and optionally FABP3, are measured in a blood, plasma, or serum sample.

[0052] In certain embodiments, biomarker measurements are obtained by a method selected from the group consisting of immunoassays, enzyme activity assays, fluorescence detection, chemiluminescence detection, electrochemiluminescence detection and patterning arrays, reverse transcriptase polymerase chain reaction, antibody binding, fluorescence activated sorting, detectable bead sorting, antibody arrays, microarrays, enzyme arrays, receptor binding arrays, allele-specific primer extension, target-specific primer extension, solid phase binding arrays, liquid phase binding arrays, fluorescence resonance transfer, and radiolabeling. In some embodiments, biomarker measurements are obtained by electrochemiluminescence detection.

[0053] In another aspect, the disclosure relates to a method for excluding a subject from recruitment into a clinical trial of an investigational Parkinson's disease drug, the method comprising the steps of: (a) obtaining a blood, plasma, or serum sample from the subject; (b) measuring in the blood, plasma, or serum sample the expression level of one or more biomarkers selected from the group consisting of IL-7, TNFα, IL-5, IL-6, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, FABP3, IL18, B2M, SAA, tenascin-C, TNRF1, and optionally PPY; (c) comparing the expression level of the one or more biomarkers in (b) to a representative statistical sample of subjects; and (d) excluding the subject from recruitment into the clinical trial if the subject is excluded from comparison with the statistical sample as having Parkinson's disease.

[0054] In some embodiments, the method further comprises the step of (e) avoiding, not initiating, or discontinuing diagnostic testing for Parkinson's disease, wherein the diagnostic testing is selected from a neurological exam, an MRI, a dopamine transporter (DAT) scan, a brain ultrasound, a PET scan, and a detailed neuropsychological test.

[0055] In some embodiments, the method further comprises (f) avoiding, not initiating, or discontinuing treatment for Parkinson's disease, wherein the treatment is selected from levodopa, dopamine agonists, glutamate agonists, anticholinergics, catechol-o-methyltransferase (COMT) inhibitors, monoamine oxidase type B (MAO-B) inhibitors, dopamine therapy, N-methyl-D-aspartate (NMDA) antagonists, and any combination thereof.

[0056] In some embodiments, step (d) is replaced with excluding the subject from further testing for recruitment into the clinical trial if the subject is excluded from comparison to the statistical sample as having Parkinson's disease. In some embodiments, the step of excluding the subject from further testing for recruitment into the clinical trial if the subject is excluded from comparison to the statistical sample as having Parkinson's disease is followed by the step of excluding the subject from recruitment into the clinical trial if the subject is excluded from comparison to the statistical sample as having Parkinson's disease.

[0057] In certain embodiments, the subject is an elderly subject. In other embodiments, the subject is a middle-aged subject. In some embodiments, the subject has concerns about changes in movement (self-reported, doctor-reported, or otherwise) or a family history of PD. In certain embodiments, the middle-aged subject is 30 years of age or older. In certain embodiments, the subject is a human subject.

[0058] In certain embodiments, the sample is a blood sample. In some embodiments, the blood, plasma, or serum sample is obtained from a subject in a primary care setting.

[0059] In some embodiments, the statistical sample comprises a group of individuals aged 30 years or older who do not have a neurological disease or disorder. In other embodiments, the statistical sample comprises a group of individuals aged 30 years or older who have been diagnosed with a neurological disease or disorder. In some embodiments, the statistical sample comprises a group of individuals aged 30 years or older who have been diagnosed with PD. In some embodiments, the statistical sample comprises a measurement of the expression level of one or more biomarkers selected from IL-7, TNFα, IL-5, IL-6, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, FABP3, IL18, B2M, SAA, tenascin-C, TNRF1, and optionally PPY from each individual in the statistical sample.

[0060] In certain embodiments, a statistically similar expression level of one or more biomarkers in a subject's sample compared to the expression levels of the corresponding biomarkers from individuals in a statistical sample without a neurological disease or disorder indicates that the subject has been ruled out as having PD and should be excluded from the clinical trial. In another embodiment, a statistically similar expression level of one or more biomarkers in a subject's sample compared to the corresponding biomarkers of individuals in a statistical sample diagnosed with PD indicates that the subject has not been ruled out as having PD and should not be excluded from the clinical trial. In some embodiments, the expression levels of one or more biomarkers measured in the subject's sample are compared to the mean expression levels of the corresponding biomarkers in the statistical sample for each group of individuals in the statistical sample.

[0061] In some embodiments, subjects who are not excluded from having PD are referred to a specialist for a diagnostic test for PD before being recruited into the clinical trial. Exemplary diagnostic tests for PD are described elsewhere herein. In some embodiments, subjects who are diagnosed by a specialist as having PD are recruited into the clinical trial, but subjects who are determined by a specialist not to have PD are not recruited into the clinical trial.

[0062] In some embodiments, the step of measuring the expression level of one or more biomarkers in the blood, plasma, or serum sample comprises measuring the expression level of one or more biomarkers of tenascin-C; tenascin-C and IL-6; tenascin-C, IL-6, and I309; tenascin-C, IL-6, I309, and IL-7; tenascin-C, IL-6, I309, IL-7, and FABP; tenascin-C, IL-6, I309, IL-7, FABP3, and TARC; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, and TNFα; tenascin-C, IL- tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, and TPO; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, and A2M; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, and IL-5; Tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, and factor VII; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, and IL-10; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, and sVCAM-1; tenascin-C, IL-6, I309, IL-7, FABP3 P3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, eotaxin 3, and SAA; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3;Tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, and IL-18; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3; tenascin-C, IL-6, I309, IL-7, FABP In some embodiments, the method comprises measuring the expression levels of PD-1, PD-2, PD-L1, PD-L2, PD-L3, TARC, TNFα, A2M, TPO, B2M, IL-5, Factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, IL-18, and sICAM-1; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, Factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, IL-18, sICAM-1, and CRP. In some embodiments, the expression levels of each of the measured biomarkers are compared with the expression levels of the corresponding biomarkers in a statistical sample to determine whether the subject should be excluded from the clinical trial as having PD.

[0063] In certain embodiments, the expression levels of one or more of tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M are measured in a blood or serum sample. In certain embodiments, the expression levels of each of tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M are measured in a blood, plasma, or serum sample. In certain embodiments, the expression levels of each of tenascin-C, IL-6, I309, and IL-7, and optionally FABP3, are measured in a blood, plasma, or serum sample.

[0064] In certain embodiments, biomarker measurements are obtained by a method selected from the group consisting of immunoassays, enzyme activity assays, fluorescence detection, chemiluminescence detection, electrochemiluminescence detection and patterning arrays, reverse transcriptase polymerase chain reaction, antibody binding, fluorescence activated sorting, detectable bead sorting, antibody arrays, microarrays, enzyme arrays, receptor binding arrays, allele-specific primer extension, target-specific primer extension, solid phase binding arrays, liquid phase binding arrays, fluorescence resonance transfer, and radiolabeling. In some embodiments, biomarker measurements are obtained by electrochemiluminescence detection.

[0065] In yet another aspect, the disclosure provides a method of screening a subject to determine whether the subject excludes having Parkinson's disease, comprising the steps of: (a) obtaining a blood, plasma, or serum sample from the subject; (b) detecting in the blood or serum sample any of interleukin (IL)-7, tumor necrosis factor alpha (TNFα), IL-5, IL-6, C-reactive protein (CRP), IL-10, soluble intercellular adhesion molecule (sICAM-1), Factor VII, I309, alpha-2-microglobulin (A2M), chemokine (CC motif) ligand 17 (TARC), eotaxin 3, soluble vascular cell adhesion molecule 1 (sVCAM-1), thrombopoietin (TPO), fatty acid binding protein (FABP), IL-18, beta-2-microglobulin (B2M), serum amyloid A1 clonogenic factor (ALK), IL-19, IL-20, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-24, IL-25, IL-26, IL-27, IL-28, IL-29, IL-30, IL-31, IL-32, IL-33, IL-34, IL-35, IL-36, IL-37, IL-38, IL-39, IL-40, IL-41, IL-42, IL-43, IL-44, IL-45, IL-46, IL-47, IL-48, IL-49, IL-50, IL-51, IL-52, IL-53, IL-54 (c) measuring the expression levels of one or more biomarkers selected from the group consisting of rastor (SAA), tenascin-C, TNRF1, and any pancreatic polypeptide (PPY); (c) comparing the expression levels of the one or more biomarkers in (b) with a representative statistical sample of subjects, thereby determining whether the subject can be ruled out or not ruled out as having Parkinson's disease; and (d) excluding subjects who are ruled out as having Parkinson's disease from a diagnostic test for Parkinson's disease, a treatment for Parkinson's disease, or a combination thereof; or (e) administering a diagnostic test for Parkinson's disease, a treatment for Parkinson's disease, or a combination thereof to subjects who are not ruled out as having Parkinson's disease.

[0066] In certain embodiments, the subject is an elderly subject. In other embodiments, the subject is a middle-aged subject. In some embodiments, the subject has concerns about changes in movement (self-reported, doctor-reported, or otherwise) or a family history of PD. In certain embodiments, the middle-aged subject is 30 years of age or older. In certain embodiments, the subject is a human subject.

[0067] In certain embodiments, the sample is a blood sample. In some embodiments, the blood, plasma, or serum sample is obtained from a subject in a primary care setting.

[0068] In some embodiments, the statistical sample comprises a group of individuals aged 30 years or older who do not have a neurological disease or disorder. In other embodiments, the statistical sample comprises a group of individuals aged 30 years or older who have been diagnosed with a neurological disease or disorder. In some embodiments, the statistical sample comprises a group of individuals aged 30 years or older who have been diagnosed with PDf. In some embodiments, the statistical sample comprises a measured expression level for one or more biomarkers selected from IL-7, TNFα, IL-5, IL-6, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, FABP3, IL18, B2M, SAA, tenascin-C, TNRF1, and optionally PPY from each individual in the statistical sample.

[0069] In certain embodiments, if the expression level of one or more biomarkers in a subject's blood, plasma, or serum sample is not statistically similar to the expression level of one or more corresponding biomarkers obtained from a group of individuals in the statistical sample who do not have Parkinson's disease, the subject is not ruled out as having Parkinson's disease. In another embodiment, if the expression level of one or more biomarkers in a subject's blood, plasma, or serum sample is statistically similar to the expression level of one or more corresponding biomarkers obtained from a group of individuals in the statistical sample who have been diagnosed with Parkinson's disease, the subject is not ruled out as having Parkinson's disease. In some embodiments, the expression level of one or more biomarkers measured in the subject's sample is compared to the mean expression level of the corresponding biomarkers in the statistical sample for each group of individuals in the statistical sample.

[0070] In some embodiments, the method further comprises the step of referring subjects who are not ruled out as having Parkinson's disease to a PD specialist for diagnostic testing.

[0071] In some embodiments, the step of measuring the expression level of one or more biomarkers in the blood, plasma, or serum sample comprises measuring the expression level of one or more biomarkers of tenascin-C; tenascin-C and IL-6; tenascin-C, IL-6, and I309; tenascin-C, IL-6, I309, and IL-7; tenascin-C, IL-6, I309, IL-7, and FABP3; tenascin-C, IL-6, I309, IL-7, FABP3, and TARC; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, and TNFα; tenascin-C, IL -6, I309, IL-7, FABP3, TARC, TNFα, and A2M; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, and TPO; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, and A2M; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, and IL-5; Tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, and factor VII; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, and IL-10; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, and sVCAM-1; tenascin-C, IL-6, I309, IL-7, FABP3 P3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, eotaxin 3, and SAA; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3;Tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, and IL-18; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, factor VII, IL-10, sVCAM-1, and eotaxin 3; tenascin-C, IL-6, I309, IL-7, FABP In some embodiments, the method comprises measuring the expression levels of biomarkers: IL-3, TARC, TNFα, A2M, TPO, B2M, IL-5, Factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, IL-18, and sICAM-1; tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, B2M, IL-5, Factor VII, IL-10, sVCAM-1, eotaxin 3, SAA, IL-18, sICAM-1, and CRP. In some embodiments, the expression levels of the biomarkers measured are compared with the expression levels of the corresponding biomarkers in a statistical sample to determine whether the subject should be excluded from a diagnostic test for Parkinson's disease.

[0072] In certain embodiments, the expression levels of one or more of tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M are measured in a blood, plasma, or serum sample. In certain embodiments, the expression levels of each of tenascin-C, IL-6, I309, IL-7, FABP3, TARC, TNFα, A2M, TPO, and B2M are measured in a blood, plasma, or serum sample. In certain embodiments, the expression levels of each of tenascin-C, IL-6, I309, and IL-7, and optionally FABP3, are measured in a blood, plasma, or serum sample.

[0073] In certain embodiments, biomarker measurements are obtained by a method selected from the group consisting of immunoassays, enzyme activity assays, fluorescence detection, chemiluminescence detection, electrochemiluminescence detection and patterning arrays, reverse transcriptase polymerase chain reaction, antibody binding, fluorescence activated sorting, detectable bead sorting, antibody arrays, microarrays, enzyme arrays, receptor binding arrays, allele-specific primer extension, target-specific primer extension, solid phase binding arrays, liquid phase binding arrays, fluorescence resonance transfer, and radiolabeling. In some embodiments, biomarker measurements are obtained by electrochemiluminescence detection.

[0074] In some embodiments, the diagnostic test is selected from the group consisting of MRI, dopamine transporter (DAT) scan, brain ultrasound, PET scan, detailed neuropsychological testing, and any combination thereof. In some embodiments, the treatment is selected from the group consisting of levodopa, dopamine agonists, glutamate agonists, anticholinergics, catechol-o-methyltransferase (COMT) inhibitors, monoamine oxidase type B (MAO-B) inhibitors, dopamine therapy, N-methyl-D-aspartate (NMDA) antagonists, and any combination thereof.

[0075] kit In yet another aspect, the present disclosure relates to a kit for screening a subject to determine whether the subject can be ruled out as having Parkinson's disease.

[0076] In certain embodiments, the kit includes a syringe and needle for use in obtaining a blood, plasma, or serum sample from a subject. In certain embodiments, the kit includes instructions.

[0077] In certain embodiments, the kit is intended for use in a primary care setting. In certain embodiments, the kit includes one or more reagents comprising a detectable marker for use in electrochemiluminescence detection. In some embodiments, the one or more reagents comprising a detectable marker are adapted for use in a multiplex biomarker assay platform using electrochemiluminescence. In certain embodiments, the detectable marker is used to measure the expression level of one or more biomarkers described elsewhere herein in a blood, serum, or plasma sample. In certain embodiments, the kit includes a "normal" blood, plasma, or serum sample to compare the expression level of one or more biomarkers in the subject sample with a "normal" sample. In certain embodiments, the kit includes a blood, plasma, or serum sample from an individual diagnosed with PD to compare the expression level of one or more biomarkers from the subject sample with a PD sample. In another embodiment, the kit includes an algorithm or a code segment comprising an algorithm for execution on a suitable processor. In certain embodiments, the algorithm compares the measured expression level of each biomarker to a representative statistical sample of subjects. In certain embodiments, the statistical sample is described elsewhere herein. In certain embodiments, the kit includes instructions on how to determine whether a subject can be ruled out as having Parkinson's disease. In another embodiment, the kit includes a second code segment executable on a suitable processor, wherein the second code segment determines whether a subject can be ruled out as having Parkinson's disease.

[0078] In yet another aspect, the present disclosure relates to kits for use as direct-to-consumer (DTC) products. In certain embodiments, the DTC kits include instructions that allow the subject to self-inquire with their primary care physician to determine whether the subject can be ruled out as having PD. In certain embodiments, the instructions include a list of common early warning signs of PD that the subject can use to self-inquire with their primary care physician. In certain embodiments, a subject with one or more warning signs of PD is instructed to visit their primary care physician to determine whether the subject can be ruled out as having PD. In certain embodiments, the methods disclosed elsewhere herein are used to determine whether a subject can be ruled out as having PD.

[0079] Experimental Example The present invention will be further described in detail by reference to the following experimental examples. These examples are provided for illustrative purposes only and are not intended to be limiting unless otherwise specified. As such, the present invention should in no way be construed as being limited to the following examples, but rather as embracing any and all variations that become apparent as a result of the teachings provided herein.

[0080] Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the compounds of the present invention and practice the claimed methods. The following working examples, therefore, specifically point out preferred embodiments of the present invention, and are not to be construed as limiting in any way the remainder of the disclosure. [Example]

[0081] Parkinson's Disease Blood Testing for Primary Care A series of studies have demonstrated the utility of blood-based biomarkers not only for detecting PD but also for differentiating it from other neurodegenerative diseases. However, no studies have demonstrated the utility of blood-based biomarkers for ruling out PD in a primary care setting. Here, a large-scale cross-validation of a PD blood test (PDBT) for use in a primary care setting was performed.

[0082] method Participant and reference database Parkinson's Disease Data. Baseline and longitudinal assays were performed on serum samples from the previously conducted DATATOP study. The DATATOP study was a multicenter, placebo-controlled clinical trial designed to test the effects of deprenyl 10 mg / day and / or tocopherol (vitamin E) 2000 IU / day (compared to levodopa) on PD progression. A total of 656 baseline PD serum samples had the requisite data for use in the current study. An additional n=190 serum samples from PD cases were already included in the research database from PD expert evaluations. Thus, there were a total of n=846 PD cases. Cases included in this study did not have a diagnosis of PD-dementia.

[0083] Neurodegenerative Disease Blood Test Reference Database (NDRD). Complex diseases, such as neurodegenerative diseases, require consideration of multiple factors (or biological pathways) rather than just a single factor when making a diagnosis. In a previous study, a blood test for detecting specific AD was developed for use in primary care settings. This blood test was discovered and validated on the premise that considering multiple biomarkers provides a more accurate approach than any single marker. This multimarker approach has led to several in vitro diagnostic (IVD) tests being advanced to clinical use in the field of oncology. However, advancing such "algorithms" to the clinic requires an appropriate reference database, which, when combined with the algorithm itself, would actually be covered by FDA regulations as software as a medical device (SAMD). Therefore, the NDRD was developed and published (O'Bryant, SE et al., "Comparing biological markers of Alzheimer's disease across blood fractions and platforms: Comparing apples to oranges," Alzheimer's Dement. Diagnosis, Assess. Dis. Monit., 2016, 3:27-34). The NDRD contains data from n > 5000 participants across a wide range of diseases (e.g., AD, PD, DLB, controls) and blood fractions (serum and plasma). Only fully de-identified data are included in the NDRD. To be included in the NDRD, data were derived from studies that (1) conducted comprehensive cognitive assessments on all participants for accurate diagnosis, and (2) were conducted under IRB approval and where written informed consent was obtained.

[0084] Controls: Controls in the database did not have a diagnosis of neurodegenerative disease, performed neuropsychological tests within normal cognitive parameters, and reported no decline in activities of daily living. For the purposes of this study, control samples derived from serum data were utilized (n=2,291 controls).

[0085] Proteomics All serum samples were assayed using a Hamilton Robotics EasyBlood system for blood processing, aliquoting, and realiquoting. A custom Hamilton Robotics StarPlus system was utilized for all plate preparation. Proteomics assays were performed in a multiplex biomarker assay platform using electrochemiluminescence (ECL) according to previously published methods using commercially available kits. ECL technology uses labels that emit light upon electrical stimulation, which improves the sensitivity of detection for many analytes, even at very low concentrations. ECL measurements have well-established properties, requiring higher sensitivity and less volume than gold-standard conventional ELISA for most assays. Analytical performance and diagnosis of several proteins (normal cognition, mild cognitive impairment, and AD) for n > 1,300 samples across multiple cohorts have recently been reported. The assay is reliable, exhibiting excellent spike recovery, dilution linearity, coefficient of variation, and limit of detection. Inter- and intra-assay variability is excellent. An internal QC protocol was implemented in addition to the manufacturing protocol, including assaying consistent controls across batches and pooled standards across lots. A total of 500 μl of serum was utilized to assay (single specimen) the following markers: fatty acid binding protein (FABP)-3, beta2 microglobulin (B2M), pancreatic polypeptide (PPY), C-reactive protein (CRP), ICAM-1, thrombopoietin, alpha2 macroglobulin (A2M), eotaxin 3, tumor necrosis factor alpha (TNF-α), tenascin-C, interleukin (IL)-5, IL-6, IL-7, IL-10, IL-18, I-309, factor 7 (Factor VII), vascular cell adhesion molecule 1 (VCAM 1), TARC, and serum amyloid α (SAA). Over 20,000 of these assays have been performed over the past few years, with all CVs <10%, and the majority <=6%.

[0086] statistical analysis Statistical analyses were performed using R (v 3.3.3) statistical software, SPSS 24 (IBM), and SAS. Support vector machine (SVM) analysis was performed to distinguish PD cases from controls. SVM is based on the concept of a discriminant plane that defines a discriminant boundary and is a classifier method that primarily performs classification tasks by constructing a hyperplane in multidimensional space that separates cases of different class labels. Diagnostic accuracy was calculated via receiver operating characteristic (ROC) curves. Samples were randomly divided into training and test samples (70 / 30), with diagnostic accuracy derived from the test sample. Finally, to provide an estimate of the overall utility of PDBT in ruling out PD in primary care settings, the negative predictive value (NPV; the probability that a subject with a negative screening test truly does not have the disease) was calculated using a range of base rates including 2%, 5%, 10%, and 15%.

[0087] result Descriptive statistics of the sample are provided in Table 1. The mean age of the sample was 63.8 (SD=13.4). The PD group was younger, more likely to be male, and reported a higher level of education (p-value <0.001) compared to the normal control group.

[0088] [Table 1]

[0089] There were a total of 592 PD samples and 1604 control samples in the training sample. SVM was applied with 5-fold internal cross-validation within the training sample for initial analysis and internal variability. PDBT gave an AUC of 0.98 with an SN of 0.84 and an SP of 0.98 within the training set. Figure 1 shows the overall classification accuracy (correct and incorrect) along with diagnostic accuracy statistics and variable importance plots.

[0090] Next, PDBT was directly applied to a test sample consisting of n = 254 PD cases and n = 687 controls. PDBT gave an AUC of 0.964 with an SN of 0.79 and an SP of 0.97. Figure 2 provides the classification accuracy (correct and incorrect) and the ROC curve.

[0091] Finally, to provide insight into how the PDBT performs as a screening tool to rule out PD in primary care settings, the NPV was calculated for a range of base rates. At a 2% base rate, the NPV was 0.99. Thus, physicians would be 99% accurate in ruling out PD with negative blood tests. The NPVs for 5%, 10%, and 15% base rates were 99%, 98%, and 96%, respectively. If physicians use a 5% base rate for adults reporting new-onset motor changes and see 5,000 patients, the PDBT would exclude 4,660 patients from the need for any additional testing procedures. There would be only 53 false-negative cases.

[0092] Selected arguments The data demonstrate the utility of the PDBT as a screen to rule out PD in a primary care setting. In the current study, data were pooled for a total of 846 PD samples and 2,291 control samples. Overall, the accuracy of the PDBT is excellent (i.e., >98%) for ruling out disease. The goal of screening tests in primary care settings for neurodegenerative diseases is to rule out disease, which is consistent with the use and performance of most screening tests routinely used in primary care settings.

[0093] The COU for PDBT is not a diagnostic but rather a screening tool to rule out PD within the primary care setting. The availability of PDBT for screening purposes in primary care holds tremendous benefits. First, it is a rapid, scalable technology that can be implemented globally as a laboratory-developed test (LTD). PDBT provides primary care providers with practical, objective information supported by several studies and many patients. Additionally, earlier treatment would likely be more beneficial for patients. The availability of PDBT in primary care settings provides a tool for rapid referral. Finally, for clinical trials, PDBT offers a means to dramatically expand access to screening procedures, far superior to specialty clinics. Overall, the current results strongly support the utility of PDBT for COU to rule out PD in primary care settings.

[0094] Those skilled in the art will readily recognize that the steps of the various above-described methods can be performed by a programmed computer. Some embodiments are also intended to encompass a program storage device, such as a digital data storage medium, which is machine- or computer-readable and encodes a machine- or computer-executable program of instructions, where the instructions perform some or all of the steps of the above-described methods. The program storage device may be, for example, a digital memory, a magnetic storage medium, such as a magnetic disk and tape, a hard drive, or any readable digital data storage medium. Some embodiments are also intended to encompass a computer programmed to perform the steps of the above-described methods.

[0095] The functions of the various elements shown in the figures, including any functional blocks labeled as "modules," may be provided through the use of dedicated hardware and hardware capable of executing software in association with appropriate software. If provided by a processor, the functions may be provided by a single dedicated processor, by a single shared processor, or by multiple individual processors, some of which may be shared. Additionally, the explicit use of the term "module" should not be construed to refer exclusively to hardware capable of executing software and may implicitly include, but is not limited to, digital signal processor (DSP) hardware, network processors, application-specific integrated circuits (ASICs), field-programmable gate arrays (FPGAs), read-only memory (ROM) for storing programs, random access memory (RAM), and non-volatile storage. Other hardware, conventional and / or custom, may also be included.

[0096] It is contemplated that any embodiment discussed herein can be implemented with respect to any method, kit, reagent, or composition of the invention, and vice versa. Furthermore, the compositions of the invention can be used to achieve the methods of the invention.

[0097] It will be understood that the specific embodiments described herein are shown by way of illustration and not as limitations of the invention. The principal features of this invention can be employed in various embodiments without departing from the scope of the invention. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific procedures described herein. Such equivalents are considered to be within the scope of this invention and are covered by the claims.

[0098] All publications and patent applications mentioned in this specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0099] The use of the words "a" or "an," when used in conjunction with the word "comprising" in the claims and / or this specification, can mean "one," but is also consistent with the meanings of "one or more," "at least one," and "one or more than one." The use of the word "or" in the claims is used to mean "and / or" unless expressly stated to refer to alternatives only or that the alternatives are mutually exclusive, but this disclosure supports a definition that refers to alternatives only and "and / or." Throughout this specification, the term "about" is used to indicate that a value includes the inherent variation of error for the device, method being employed to determine the value, or the variation that exists among study subjects.

[0100] As used in the specification and claims, the words "comprising" (and any form of comprising, e.g., "comprise" and "comprises"), "having" (and any form of having, e.g., "have" and "has"), "including" (and any form of including, e.g., "includes" and "include"), or "containing" (and any form of containing, e.g., "contains" and "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. In any of the embodiments of the compositions and methods provided herein, "comprising" may be substituted with "consisting essentially of" or "consisting of." As used herein, the phrase "consisting essentially of" requires the specified integers or steps and those that do not materially affect the properties or function of the claimed invention. As used herein, the term "consisting of" is used to indicate only the presence of the enumerated integers (e.g., features, elements, properties, properties, method / process steps, or limitations) or group of integers (e.g., features, elements, properties, properties, method / process steps, or limitations).

[0101] As used herein, the term "or combinations thereof" refers to all permutations and combinations of the listed items preceding the term. For example, "A, B, C, or combinations thereof" is intended to include at least one of A, B, C, AB, AC, BC, or ABC, and, where order is important in a particular context, also BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, combinations containing one or more repeats of an item or term are expressly included, e.g., BB, AAA, AB, BBC, AAABCCCC, CBBAAA, CABABB, etc. Those of skill in the art will understand that there is typically no limit to the number of items or terms in any combination, unless otherwise clear from the context.

[0102] As used herein, terms of approximation, such as, but not limited to, "about," "substantial," or "substantially," when so modified, refer to conditions that are understood not to be necessarily absolute or complete, but are believed to be sufficiently close to those of ordinary skill in the art to warrant specification of the conditions being presented. The degree to which the description may be varied depends on what magnitude of change may be made, and one of ordinary skill in the art will still recognize the modified feature as still possessing the required properties and capabilities of the unmodified feature. Generally, subject to the preceding discussion, numerical values ​​herein modified by terms of approximation such as "about" may vary by at least ±1, 2, 3, 4, 5, 6, 7, 10, 12, or 15% from the stated value.

[0103] Additionally, section headings herein are provided for consistency with the implications of 37 CFR 1.77 or to otherwise provide organizational guidance. These headings do not limit or characterize the invention(s) set forth in any claim that may issue from this disclosure. Specifically, and by way of example, a heading may refer to a "Field of the Invention," but such claim(s) should not be limited by language under this heading describing a so-called technical field. Furthermore, the description of a technology in the "Background of the Invention" section should not be construed as an admission that the technology is prior art to any invention(s) in this disclosure. Nor should the "Summary" be considered a characterization of the invention(s) set forth in any issued claim. Furthermore, any reference to the singular "invention" in this disclosure should not be used to assert that there is only a single novel feature in this disclosure. Multiple inventions may be set forth according to the limitations of multiple claims that issue from this disclosure, and such claims therefore define the invention(s) and their equivalents protected thereby. In all instances, the scope of such claims should be considered on their own merits in light of this disclosure, but should not be constrained by the headings set forth herein.

[0104] All of the compositions and / or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those skilled in the art that variations can be applied to the compositions and / or methods, and to the steps or sequence of steps of the methods described herein, without departing from the concept, spirit, and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope, and concept of the invention as defined by the appended claims.

[0105] To assist the Patent Office, and any reader of any patent that may issue on this application, in interpreting the claims appended hereto, Applicants wish to note that unless the words "means for" or "step for" are expressly used in a particular claim, they do not intend that any of the appended claims invoke 35 U.S.C. § 112(6), 112(f), or their equivalents, as the appended claims exist on the filing date thereof.

[0106] For each claim, each dependent claim may depend on both the independent claim and the preceding dependent claim, so long as the preceding claim provides suitable antecedent for a term or element of the claim, for each and every claim.

Claims

1. 1. A method of collecting data to exclude a subject from the need for diagnostic testing for Parkinson's disease, comprising: (a) measuring the expression levels of the biomarkers tenascin-C and interleukin (IL)-7 in a blood, plasma, or serum sample from the subject; (b) comparing the expression levels of the biomarkers in (a) to a representative statistical sample of the subjects; (c) collecting data to determine whether the subject can be ruled out as having Parkinson's disease based on the measured biomarkers; and (d) collecting data to determine that the subject should be excluded from diagnostic testing for Parkinson's disease based on the comparing step. The method comprising:

2. 10. The method of claim 1, further comprising the step of: (e) collecting data to avoid, not initiate, or discontinue diagnostic testing for Parkinson's disease, wherein the diagnostic testing is selected from the group consisting of a neurological exam, an MRI, a dopamine transporter (DAT) scan, a brain ultrasound, a PET scan, a detailed neuropsychological test, and any combination thereof.

3. 10. The method of claim 1, further comprising the step of: (e) collecting data to avoid, not initiate, or discontinue treatment for Parkinson's disease, wherein the treatment is selected from the group consisting of levodopa, dopamine agonists, glutamate agonists, anticholinergics, catechol-o-methyl transferase (COMT) inhibitors, monoamine oxidase type B (MAO-B) inhibitors, dopamine therapy, N-methyl-D-aspartate (NMDA) antagonists, and any combination thereof.

4. (a) is a measurement of the presence or absence of tumor necrosis factor alpha (TNFα), IL-5, IL-6, C-reactive protein (CRP), IL-10, soluble intracellular adhesion molecule (sICAM-1), factor VII, I309, alpha-2-microglobulin (A2M), chemokine (CC motif) ligand 17 (TARC), eotaxin 3, soluble vascular cell adhesion molecule 1 (sVCAM-1), thrombopoietin (TPO), fatty acid binding protein (FABP) in a blood, plasma, or serum sample derived from a subject; 10. The method of claim 1, further comprising measuring the expression level of one or more additional biomarkers selected from the group consisting of IL-18, beta-2-microglobulin (B2M), serum amyloid Al cluster (SAA), tumor necrosis factor receptor 1 (TNFR1), and pancreatic polypeptide (PPY).

5. 5. The method of claim 4, wherein the one or more additional biomarkers are selected from the group consisting of IL-6, I309, and FABP.

6. 2. The method of claim 1, wherein (a) further comprises the step of measuring the expression level of each biomarker in the group consisting of IL-6 and I309 in a blood, plasma, or serum sample from the subject.

7. 7. The method of claim 6, further comprising measuring the expression level of one or more additional biomarkers selected from the group consisting of FABP, TNFα, IL-5, CRP, IL-10, sICAM-1, Factor VII, I309, A2M, TARC, eotaxin 3, sVCAM-1, TPO, IL-18, B2M, SAA, and PPY in a blood, plasma, or serum sample from the subject.

8. 10. The method of claim 1, wherein the expression level of the biomarker is measured using electrochemiluminescence.

9. (d) collecting data (i) collecting data to determine if the expression levels of the biomarkers in (a) are statistically similar to the mean expression levels of the corresponding biomarkers obtained from a group of individuals in a statistical sample who do not have Parkinson's disease; or (ii) collecting data to determine if the expression level of the biomarker in (a) is not statistically similar to the mean expression level of the corresponding biomarker obtained from a group of individuals in the statistical sample who have been diagnosed with Parkinson's disease. The method of claim 1 , comprising:

10. (e) collecting data to exclude a subject from recruitment into a clinical trial if the subject is excluded from comparison to the statistical sample as having Parkinson's disease. The method of claim 1 further comprising:

11. (i) collecting data to exclude subjects who are excluded from having Parkinson's disease from diagnostic testing for Parkinson's disease, treatment for Parkinson's disease, or a combination thereof; or (ii) collecting data to administer a diagnostic test for Parkinson's disease, a treatment for Parkinson's disease, or a combination thereof to subjects who are not excluded from having Parkinson's disease. The method of claim 1 further comprising:

12. (b) using a computer to compare the expression levels of the biomarkers in (a) to a representative statistical sample of subjects; (c) using a computer to collect data to determine whether the subject can be ruled out as having Parkinson's disease; and (d) using a computer to collect data for determining that the subject should be excluded from diagnostic testing for Parkinson's disease based on the comparing step. The method of claim 1.

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