WO2018129421A1 - A promising drug candidate for parkinson's disease - Google Patents

Abstract

The purpose of this study and report was to evaluate GM6 as a Parkinson's Disease (PD) drug candidate and the possible mechanism of action and pathways. We show that GM6 rescues in vitro survival of salsolinol-treated SH-SY5Y cells and rat cortical neurons treated with post-mortem CSP from PD (Parkinson's Disease) patients. We show that short-term GM6 treatment improves BDNF blood levels i PD patients. Additionally, GM6 completely or partially abrogated dysfunction in 2 in vivo PD mouse models (6-0HDA, MPTP), leading to improved motor performance and dopaminergic neuron survival in both models. We also identify PD-associated genes regulated by GM6. These genes were categorized with respect to 6 hypothesized mechanisms of action, including (1 ) attenutation of kinase dysfunction via LRKK2 down-regulation, (2) anticholinergic actions, (3) pro-GABAergic effects, (4) enhancement of GDNF activity through activation of a GRFAI-RET axis, (5) improved oxidative stress defenses and (6) blunting of free radical generation through decreased mitochondrial activity or abundance.

Description

TITLE

A PROMISING DRUG CANDIDATE FOR PARKINSON'S DISEASE

FIELD

The field includes using the MNTF Factor known as GM6 for the diagnosing, monitoring, prognosing Parkinson's disease, and preventing, delaying the onset, or treating Parkinson's disease through regulation of expression of multiple genes by GM6 to regulate the pathology of Parkinson's disease.

RELATED APPLICATIONS

This Application takes priority from U.S.S.N. 62/444,032, filed January 9, 2017, entitled 'GM6 is a regulator of Parkinson's Disease associated genes and regulatory pathways and is a promising drug candidate for Parkinson's Disease', by Pui-Yuk Dorothy o, incorporated by reference in its' entirety.

BACKGROUND

The following includes information that may be useful in understanding the present inventions. It is not an admission that any of the information provided herein is prior art, or relevant, to the presently described or claimed inventions, or that any publication or document that is specifically or implicitly referenced is prior art.

The isolation and characterization of two motoneuronotrophic factors (MNTF1 and MNTF2) from rat muscle tissues, as well as the subsequent cloning of a recombinant MNTF1-F6 gene derived from a human retinoblastoma cDNA library, is previously described in Applicant's prior U.S. Patent No. 6,309,877. The MNTF1-F6 gene sequence encodes a 33 amino acid sequence referred to herein as SEQ ID NO:l having the following amino acid sequence: LGTFWGDTLNCWMLSAFSRYARCLAEGHDGPTQ [SEQ ID NO: 1].

The naturally occurring and recombinant MNTF1 polypeptides were shown to selectively enhance the survival in vitro of anterior horn motor neurons isolated from rat lumbar spinal cord explants. Photomicrographs of treated cultures exhibited neurite outgrowth of myelinated nerve fibers and a marked reduction in the growth of non-neuronal cells, e.g. glial cells and fibroblasts. Similarly, in vivo administration of MNTF1 to surgically axotomized rat peripheral nerves resulted in a markedly higher percentage of surviving motor neurons than untreated controls, which could be blocked by co-administration of anti-MNTFl monoclonal antibody. Further beneficial effects of MNTF1 were demonstrated in rats subjected to spinal cord hemi-section, repaired by a peripheral nerve autograft and implanted with MNTF1- containing gel sections in close proximity to the nerve graft junctions with spinal cord. MNTF1 treated animals exhibited greater numbers of surviving motor neurons, improved recovery of motor and sensory function, reduced inflammatory response (fewer infiltrating macrophages and lymphocytes) and reduced collagen-containing scar tissue formation at the site of the graft, normal Schwann cell morphology and normal myelinated and non-myelinated nerve fiber formation.

Two previously unrecognized overlapping domains within the MNTF1-F6 molecule that appear to be sufficient for the known biological activities of MNTF1 have now been identified. Each of these domains, designated herein as the "WMLSAFS" and "FSRYAR" domains, are sufficient to stimulate the proliferation of motor neuron derived cell lines in a manner similar to the MNTF1-F6 33-mer. Similarly, the "FSRYAR" domain is sufficient to direct selective reinnervation of muscle targets by motor neurons in vivo in a manner similar to the MNTF1-F6 33-mer. In addition, the "FSRYAR" domain provides an antigenic epitope sufficient to raise antibody that recognizes any MNTF peptide containing the "FSRYAR" sequence, including the MNTF1-F6 33-mer.

Novel peptides and composition from active fragments of MNTF that are capable of modulating viability and growth in neuronal cells, and to methods of modulating neuronal cell viability and growth employing the novel peptides and compositions, containing either a "WMLSAFS domain" or "FSRYAR domain", which is sufficient for neurotrophic or neurotropic function is described in US Patent 7,183,373. The polypeptide domain demonstrated therein were sufficient for the selective maintenance and axonal regeneration of neuronal cells, and to peptides and/or molecules capable of mimicking their structure and/or function. Preferred embodiments of that invention comprise a peptide having the amino acid sequence: FSRYAR [SEQ ID NO:2], the sequence of GM6, also known as GM604 for ALS indication, as well as analogues thereof. Preferably such analogues are functional equivalents of the GM6 [SEQ ID NO:2].

GM6 encompasses the active domain of MNTF, an endogenous master neural growth regulator present during the fetal development phase when neurons are being created and reaching terminal synaptic targets. The fetal phase is the most intense and rapid period of human growth and development, especially within the CNS. In preclinical studies, treatment of GM604 in rodents demonstrated that it promotes neural regeneration and exhibits both trophic and tropic effects (Chau RMW et al. 1990 Neuronotrophic Factor. Chin J. Neuroanat. 6:129-138; Chau RMW et al. 1992. Muscle neurotrophic factors specific for anterior horn motoneurons of rat spinal cord. Recent Adv. Cell Mol Biol. 5:89-94; Yu J. et al. 2008. Motoneuronotrophic Factor analog GM604 reduces infarct volume and behavioral deficits following transient ischemia in the mouse. Brain Res. 1238:143-153; US 7,183,373).

Parkinson's disease (PD) or paralysis agitans is a basal ganglia disorder characterized by the loss of dopaminergic neurons within the midbrain substantia nigra pars compacta and ventral tegmental area. It is a relatively common degenerative disease of aging affecting approximately 1% of the population 50 years of age and older. The disease is usually diagnosed based upon clinical symptoms although radioactive positron emission tomography can be used as a diagnostic aide. Clinical symptoms of PD include a "pill rolling" tremor at rest, cogwheel or lead-pipe rigidity, masked (expressionless) facies, restlessness (akinesia), bradykinesia, postural instability, and shuffling gait with festination and loss of arm swing. Patients may additionally present with sialorrhea (excess of saliva) and display micrographia (small handwriting). In contrast to other forms of tremor, the PD resting tremor is most noticeable in the hands and distal appendages and is alleviated by intentional movement. Histologically, PD is associated with the loss of neuromelanin pigment granules and the presence of intracellular eosinophilic alpha- synuclein inclusions known as Lewy bodies. It is likely that both genetic and environmental factors contribute to PD. The genetic component is polygenic and thus far more than 41 genetic loci have been linked to PD based upon genome- wide association studies (PMID: 28892059).

PD is driven by decreased activity of the nigrostriatal dopamine pathway secondary to the loss of dopaminergic neurons leading to decreased dopamine abundance in the caudate nucleus and putamen. However, disturbance of other neurotransmitters is also observed, with decreased abundance of serotonin and elevations of acetylcholine, leading to altered neurotransmitter balance that may worsen or contribute to the disease process (PMID: 20590830). Additionally, dopaminergic neurons have been observed to also produce the inhibitory neurotransmitter gamma- Aminobutyric acid (GAB A) (PMID: 23034651), and correspondingly GABA is reduced in the prefrontal cortex of PD patients (PMID: 20832408), which has prompted formulation of the "GABA collapse" hypothesis suggesting that GABA deficiency may be of similar importance to dopamine in PD pathogenesis (PMID: 27375426)

At present, a range of pharmaceutical options have been approved to manage PD and control symptoms, including agents increasing L-DOPA availability and preventing its degradation (L-DOPA/carbidopa), agents that prevent dopamine degradation (selegiline, tolcapone), dopamine agonists (bromocriptine, pramipexole, ropinirole), drugs increasing dopamine availability (amantadine) and anti-cholinergic drugs (benztropine, trihexyphenidyl). The mainstay of PD therapy has been the synergistic combination of Levodopa with carbidopa (Sinemet), which serves to increase L-DOPA while the addition of DOPA decarboxylase inhibitor (carbidopa) increases L-DOPA bioavailability to facilitate its transport across the blood-brain barrier. Ultimately, however, the long-term use of levodopa/carbidopa is limited by adverse effects, which include development of arrhythmias along with dyskinesia. All current PD treatments are supportive and intended to address symptoms and delay disease progression, but no existing treatment has been demonstrated to restore loss of dopamine production in the midbrain of advanced PD patients. Given that defects in PD neurotransmission may thus be broader than currently recognized and not limited to dopamine, development of new multi-target may will be needed to fully prevent functional decline in PD patients (PMID: 28328536).

GM6 is an oligopeptide containing 6 amino acids, corresponding to the active site of an embryonic-stage protein functioning within the developing nervous system. GM6 has been shown to cross the blood-brain barrier (Yu et al. 2008, Brain Res. 1238: 143-153). The safety and tolerability of GM6 has been demonstrated in phase 1 and phase 2 clinical trials in Parkinson's Disease (PD), Amyotrophic Lateral Sclerosis (ALS) and Ischemic stroke. It has been proposed that GM6 triggers activation of pro-survival pathways mediating neural development, which may in turn have beneficial effects for treatment of multiple neurodegenerative diseases.

The purpose of this study was to provide a bioinformatic analysis to assess whether GM6 alters the expression of genes associated with PD. Gene expression analyses are performed using several gene expression profiling datasets generated using DNA microarray or RNA-seq technology. These datasets evaluated effects of GM6 on gene expression in SH-SY5Y neuroblastoma cells. Effects of GM6 in SH-SY5Y neuroblastoma cells were of main interest as an in vitro dopaminergic neuron model (PMID: 20497720). Goals were to determine whether PD-associated genes exhibit unique responses to GM6 treatment, which would suggest that GM6 is impacting signaling pathways linked to core processes that underlie PD onset and/or progression.

BRIEF SUMMARY

The inventions described and claimed herein have many attributes and embodiments including, but not limited to, those set forth or described or referenced in this Summary. The inventions described and claimed herein are not limited to or by the features or embodiments identified in this Summary, which is included for purposes of illustration only and not restriction.

These and other aspects and embodiments of the inventions described and claimed herein will be apparent from and throughout the application and claims, all of which shall be considered to be a part of the written description thereof. The inventions provided herein are directed toward methods of diagnosing, monitoring, prognosing, and preventing, delaying the onset, or treating Parkinson's Disease (PD).

Accordingly, one embodiment is directed to a method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide with the peptide sequence FSRYAR (GM6) to a subject to regulate the pathology of Parkinson's disease, wherein GM6 regulates more than one downstream target to regulate the pathology of the disease. There are numerous variations of this embodiment, including without limitation those where: GM6 acts as a master regulator of multiple targets to regulate the pathology of Parkinson' s disease to treat or prevent Parkinson's disease, where GM6 functions both as an agonist and an antagonist for different targets that regulate the pathology of Parkinson's disease, and where GM6 functions both as an agonist and an antagonist for different targets that regulate the pathology of Parkinson's disease, wherein GM6 regulates the pathology of Parkinson's disease by up regulating some targets and downregulating other targets to achieve a homeostasis. Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising the administering a MNTF peptide consisting of the amino acids FSRYAR (GM6) to a subject to regulate the pathology of Parkinson's disease, wherein GM6 regulates more than one downstream target to inhibit mitochondria-mediated neuronal cell death associated with Parkinson' s disease. Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide with the peptide sequence FSRYAR (GM6) to a subject, wherein GM6 regulates more than one of the following downstream targets to regulate the pathology of the disease: GNAI1, CASP3, PRKACB, UBE2L3, CASP9, UBE2J1, SNCAIP, ATP5B, GPR37, GNAI3, SLC18A1, UBE2G1, APAF1, UQCRC2, NDUFS1, NDUFV3, PINK1, PARK2, SNCA, NDUFA4L2, NDUFV1, ATP5E, UBE2J2, NDUFS2, UQCRC1, HTRA2, ADORA2A, COX6A2, COX7A1, LRRK2, PPIF, SLC18A2, VDAC3, SDHD, SLC25A6, COX4I2, NDUFC2, SLC25A31, GNAL, PRKACG, COX7B2, NDUFA10, SLC6A3, UQCRB, ATP5J, COX6B2, DRD2, ADCY5, UBE2L6, GNAI2, ATP5F1, UCHL1, ATP5C1, COX7A2L, UBB, SLC25A5, TH, NDUFB5, NDUFA5, UBE2G2, SDHB, CYCS, CYC1, COX4I1, NDUFB3, ATP5G3, COX8A, NDUFA9, PRKACA, UQCRQ, NDUFS3, COX6B1, NDUFA1, SLC25A4, NDUFS4, COX5B, NDUFA6, NDUFS8, ATP5D, NDUFA13, NDUFB9, NDUFS5, NDUFB10, NDUFB11, NDUFA7, NDUFS6, NDUFA8, NDUFA2, UQCRH, COX7C, NDUFB4, NDUFB6, NDUFC1, COX6C, NDUFAB1, NDUFB2, NDUFB7, COX7A2, COX7B, ATP50, COX5A, PARK7, NDUFA12, and NDUFBl.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide with the peptide sequence FSRYAR (GM6) to a subject, wherein GM6 down-regulates at least one of the following downstream targets to regulate the pathology of the disease: NDUFBl, NDUFA12, PARK7, COX5A, ATP50, COX7B, COX7A2, NDUFB7, NDUFB2, NDUFAB1, COX6C, NDUFC1, NDUFB6, NDUFB4, COX7C, UQCRH, NDUFA2, NDUFA8, NDUFS6, NDUFA7, NDUFBl 1, NDUFB10, NDUFS5, NDUFB9, NDUFA13, ATP5D, NDUFS8, NDUFA6, COX5B, NDUFS4, SLC25A4, NDUFA1, COX6B1, NDUFS3, UQCRQ, PRKACA, NDUFA9, COX8A, ATP5G3, NDUFB3, COX4I1, CYC1, CYCS, SDHB, UBE2G2, NDUFA5, NDUFB5, TH, SLC25A5, UBB, COX7A2L, and UCHL1.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide consisting of the amino acids FSRYAR (GM6) to a subject, wherein GM6 down-regulates at least one of the following downstream targets to regulate the pathology of the disease: NDUFBl, NDUFA12, PARK7, COX5A, ATP50, COX7B, COX7A2, NDUFB7, NDUFB2, NDUFAB1, COX6C, NDUFC1, NDUFB6, NDUFB4, COX7C, UQCRH, NDUFA2, NDUFA8, NDUFS6, NDUFA7, NDUFBl 1, NDUFB10, NDUFS5, NDUFB9, NDUFA13, ATP5D, NDUFS8, NDUFA6, COX5B, NDUFS4, SLC25A4, NDUFA1, COX6B1, NDUFS3, UQCRQ, PRKACA, NDUFA9, COX8A, ATP5G3, NDUFB3, COX4I1, NDUFB5, and TH.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide consisting of the amino acids FSRYAR (GM6) to a subject, wherein GM6 up- regulates at least one of the following downstream targets to regulate the pathology of the disease: GNAI1, CASP3, PRKACB, UBE2L3, CASP9, UBE2J1, SNCAIP.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide consisting of the amino acids FSRYAR (GM6) to a subject, wherein GM6 up- regulates at least one of the following downstream targets to regulate the pathology of the disease: GNAI1, CASP3, PRKACB.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide with the peptide sequence FSRYAR (GM6) to a subject, wherein GM6regulate the genes in pathways selected from the following: mitochondrial protein complex, respiratory chain complex, mitochondrial respiratory chain complex, NADH dehydrogenase complex, mitochondrial envelope, envelope, organelle membrane, catalytic complex, intracellular organelle part, mitochondrion, macromolecular complex, mitochondrial inner membrane, mitochondrial membrane part, mitochondrial respiratory chain, mitochondrial intermembrane space.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by (up) -regulating the expression of one or more genes selected from the group consisting of: MAPT; RAB29; PERI; HSPB1; SLC45A3; HIP1R; RET; CAPN1; ACHE; LINGOl; DNM1; NPTX2; HGF; BCKDK; SLC41A1; SNCAIP; NEDD9; GPR37; CNKSR3; HMOX1; MAOA; DRD2; SREBF1; GAD1; GFRA1; SEPT4; APOE; GST02; SNCB; WNT3; VEGFA; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; KRTCAP2; SLC50A1; C8orf4; CRHR1-IT1 VDAC1; NOS1; BST1; CRYAB; SOD2; HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl; TMEM229B; TGM2 and HSPA1A.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by (down) -regulating the expression of one or more genes selected from the group consisting of: NDUFS2; LMNB1; PRDM15; CD200; MRPL3; NSF; BRINP1; WNT3; ASCL1; SEMA5A; SFXN2; GRK5; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9 and COX4I2; ATXN3; MCCC1; NDUFB2; HSPA8; SIPA1L2; CAT; TP53; CNNM2; SLC25A4; PRDX3; BORCS7; ZNF646; TRAF6; SIAH1; HTT; DDRGK1; UNC13B; GIGYF2; P2RX7; NDUFS4; and LRRK2. Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by (up) -regulating the expression of one or more genes selected from the group consisting of: MAPT; SLC41A1; RAB29; PERI; SNCAIP; HSPB1; SLC45A3; HIP1R; NEDD9; RET; GPR37; CAPN1; ACHE; CNKSR3; LINGOl; DNM1; NPTX2; HMOX1; MAOA; DRD2; SREBF1; HGF; GAD1; GFRA1; SEPT4; BCKDK and APOE. Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by (down) -regulating the expression of one or more genes selected from the group consisting of: NSF; BRINP1; NDUFS2; WNT3; ASCL1; SEMA5A; LMNB1; PRDM15; SFXN2; CD200; GRK5; MRPL3; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9 and COX4I2.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by (up) -regulating the expression of one or more genes selected from the group consisting of: CAPN1; HGF; GST02; SNCB; ACHE; BCKDK; HSPB1; DNM1; WNT3; VEGFA; RAB29; HIP1R; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; SLC45A3; KRTCAP2; RET; SLC50A1; LINGOl; C8orf4; PERI; MAPT; CRHR1-IT1; VDAC1; NOS1; NPTX2; BST1; CRYAB; SOD2; HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl; TMEM229B; TGM2 and HSPA1A.

Another embodiment is a method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by (down) -regulating the expression of one or more genes selected from the group consisting of: ATXN3; MCCC1; NDUFB2; HSPA8; LMNB1; MRPL3; SIPA1L2; CAT; TP53; PRDM15; CNNM2; SLC25A4; NDUFS2; PRDX3; BORCS7; ZNF646; TRAF6; SIAH1; HTT; DDRGK1; UNC13B; GIGYF2; P2RX7; CD200; NDUFS4 and LRRK2.

Another embodiment is a method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: MAPT; RAB29; PERI; HSPB1; SLC45A3; HIP1R; RET; CAPN1; ACHE; LINGOl; DNM1; NPTX2; HGF; BCKDK; SLC41A1; SNCAIP; NEDD9; GPR37; CNKSR3; HMOX1; MAOA; DRD2; SREBF1; GAD1; GFRA1; SEPT4; APOE; GST02; SNCB; WNT3; VEGFA; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; KRTCAP2; SLC50A1; C8orf4; CRHR1- IT1 VDAC1; NOS1; BST1; CRYAB; SOD2; HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl; TMEM229B ; TGM2 and HSPA1A in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson's disease.

Another embodiment is a method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: NDUFS2; LMNB1; PRDM15; CD200; MRPL3; NSF; BRINP1; WNT3; ASCL1; SEMA5A; SFXN2; GRK5; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9 and COX4I2; ATXN3; MCCC1; NDUFB2; HSPA8; SIPA1L2; CAT; TP53; CNNM2; SLC25A4; PRDX3; BORCS7; ZNF646; TRAF6; SIAH1; HTT; DDRGK1; UNC13B; GIGYF2; P2RX7; NDUFS4; and LRRK2 in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson's disease. Another embodiment is a method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: MAPT; SLC41A1; RAB29; PERI; SNCAIP; HSPB1; SLC45A3; HIP1R; NEDD9; RET; GPR37; CAPN1; ACHE; CNKSR3; LINGOl; DNM1; NPTX2; HMOX1; MAOA; DRD2; SREBFl; HGF; GADl; GFRAl; SEPT4; BCKDK and APOE in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson' s disease.

Another embodiment is a method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: NSF; BRINP1; NDUFS2; WNT3; ASCL1; SEMA5A; LMNB1; PRDM15; SFXN2; CD200; GRK5; MRPL3; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9 and COX4I2 in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson' s disease. Another embodiment is a method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: CAPN1; HGF; GST02; SNCB; ACHE; BCKDK; HSPB1; DNM1; WNT3; VEGFA; RAB29; HIP1R; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; SLC45A3; KRTCAP2; RET; SLC50A1; LINGOl; C8orf4; PERI; MAPT; CRHR1-IT1; VDAC1; NOS1; NPTX2; BST1; CRYAB; SOD2; HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl; TMEM229B; TGM2 and HSPA1A in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson' s disease.

Another embodiment is a method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: ATXN3; MCCC1; NDUFB2; HSPA8; LMNB1; MRPL3; SIPA1L2; CAT; TP53; PRDM15; CNNM2; SLC25A4; NDUFS2; PRDX3; BORCS7; ZNF646; TRAF6; SIAH1; HTT; DDRGK1; UNC13B; GIGYF2; P2RX7; CD200; NDUFS4 and LRRK2 in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson' s disease.

There are numerous variations of the embodiment described above, including without limitation those where: GM6 is used in combination with another drug or therapy, where GM6 is administered intravenously, where GM6 is administered subcutaneously, where GM6 is administered orally, and those where GM6 is administered by combination of intravenously and subcutaneously.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 shows that GM6 prevents cell death following treatment with salsolinol or post-mortem CSF from PD patients. (A) Salsolinol. SH-SY5Y cells were untreated (CTL) or treated for 24 hours with GM6 (0.1, 1, or 10 mg/ml) or GM6 + salsolinol (100 μΜ) (n = 10/group). (B) PD post-mortem CSF. Sprague Dawley rat cortical neuronal cells were treated with post-mortem CSF from control subjects or PD patients (n = 5/group). In (A) and (B), thiazolyl blue (MTT) assays were used to assess cell viability. Groups without the same letter differ significantly (P < 0.05, Tukey HSD test).

Fig. 2 shows PD-associated genes most strongly altered by GM6 (Microarray). The figure shows PD-associated genes most strongly altered in SH-5YSY cells treated with GM6 for 48 hours. PD-associated genes with lowest p-values are shown. An asterisk (*) is used to denote genes significantly altered with FDR < 0.10 (left margin). Black bars denote genes additionally altered by GM6 based upon a FC threshold (FC > 1.50 or FC < 0.67). All genes were significantly altered by GM6 (FDR < 0.10) with FC > 1.50 or FC < 0.67. Genes were linked to PD based upon 2 or more of 9 possible database sources.

Fig. 3 illustrates PD-associated genes are predominantly repressed by GM6 in SH-SY5Y cells and associated with mitochondria. (A) Simulation analyses. Sets of 104 SH- SY5Y-expressed genes were sampled at random. The histogram shows average FC among sets of 104 randomly sampled genes (arrow: observed average FC among 104 PD-associated genes). (B) FC estimates. The proportion of GM6-decreased genes (FC < 1) was significantly larger than expected (P = 1.65 x 10-12; Fisher's exact test). (C) GO CC terms enriched among PD- associated GM6-decreased genes (conditional hypergeometric test; left margin parentheses: no. of GM6-decreased genes associated with each term; right margin: example GM6-decreased genes associated with each term).

Fig. 4 illustrates the KEGG PD pathway (hsa05012). Dark grey elements are associated with genes up-regulated in GM6-treated cells compared to CTL cells, and light grey elements are associated with genes down-regulated in GM6-treated cells compared to CTL cells (see color scale, lower right). White elements have not been assigned to specific genes. An interactive version of this diagram is available online:

http://www.genome.jp/kegg-bin/show pathway'?hsa05012

Fig. 5 illustrates that GM6 improves behavioral, biochemical, and histological features in the 6-OHDA mouse model. Male C57BL/6 mice received intracerebral injection of 6- OHDA or solvent (CTL). Treated mice received GM6 twice daily (i.v.) over 5 days after 6- OHDA treatment (0, 1, 5, 10 or 20 mg/kg in saline). At 1 week following 6-OHDA treatment, we evaluated (A) spontaneous activity, (B) rotarod performance, (C) brain dopamine, (D) brain DOPAC, (E) brain HVA, and (F) number of tyrosine hydroxylase positive neurons in the substantia nigra pars compacta (n = 10/group). Groups without the same letter differ significantly (P < 0.05, Tukey HSD test).

Fig. 6 shows how GM6 improves behavioral, biochemical, and histological features in the MPTP mouse model. C57BL/6 mice received intracerebral injection of MPTP or solvent (CTL). Treated mice received GM6 twice daily (i.v.) over 4 days after MPTP treatment (0, 1, 5, 10 or 20 mg/kg in saline). At 1 week following MPTP treatment, we evaluated (A) spontaneous activity, (B) rotarod performance, (C) brain dopamine, (D) brain DOPAC, (E) brain HVA, and (F) number of tyrosine hydroxylase positive neurons in the substantia nigra pars compacta (n = 10/group). Groups without the same letter differ significantly (P < 0.05, Tukey HSD test). Fig. 7 shows short-term GM6 treatment improves BDNF blood levels in PD patients (Phase 2A trial). PD patients received 6 doses of GM6 (n = 4) or placebo (n = 2) over 2 weeks. BDNF was evaluated at baseline (visit 1), after 4 doses (visit 4), after 6 doses (visit 6) and 10 weeks post-treatment (visit 8). We analyzed the change in BDNF blood level for each patient relative to baseline at visits 4, 6, and 8 (one-tailed paired t-test).

Fig. 8 shows genes associated with PD and their response to GM6 treatment (3+ database sources; directional test). PD-associated genes were identified and their average fold- change (GM6/CTL) was compared to randomly sampled gene sets. The arrow indicates the average fold-change (GM6/CTL) among PD-associated genes. The grey histogram represents the distribution of average fold-change estimates in randomly sampled gene sets (10,000 random samples for each analysis). PD-associated genes used for this analysis include only those linked to PD based upon at least 3 database sources.

Fig. 9 shows genes associated with PD and their response to GM6 treatment (3+ database sources; non-directional test). PD-associated genes were identified and their average absolute fold-change [2^Aabs(log2(GM6/CTL))] was compared to randomly sampled gene sets. The arrow indicates the average absolute fold-change among PD-associated genes. The grey histogram represents the distribution of average absolute fold-change estimates in randomly sampled gene sets (10,000 random samples for each analysis). PD-associated genes used for this analysis include only those linked to PD based upon at least 3 database sources.

Fig. 10 shows PD-associated genes most strongly altered by GM6 (RNA-seq).

The figure shows PD-associated genes most strongly altered in SH-5YSY cells (A) treated with GM6 for 6 hours (UM, RNA-seq), (B) treated with GM6 for 24 hours (UM, RNA-seq), (C) treated with GM6 for 48 hours (UM, RNA-seq), (D) treated with GM6 for 6 hours (SBH, 6 hours), and (E) treated with GM6 for 24 hours (SBH, 24 hours). PD-associated genes with lowest p-values are shown in each case. Genes with black bars were altered by GM6 with FC > 1.50 or FC < 0.67. An asterisk (*) is used to denote genes significantly altered with FDR < 0.10 (left margin). Genes were linked to PD based upon 2 or more of 9 possible database sources.

Fig. 11 shows Gene ontology biological process (BP) terms associated with PD- associated genes altered by GM6 (UM, RNA-seq). (A) Gene ontology BP terms enriched with respect to GM6-increased PD-associated genes (FDR < 0.10; pooled from the 3 UM RNA-seq experiments). Values in parentheses (left margin) indicate the number of GM6-increased genes in each category (right margin: exemplar genes most strongly induced by GM6). (B) Gene ontology BP terms enriched with respect to GM6-decreased PD-associated genes (FDR < 0.10; pooled from the 3 UM RNA-seq experiments). Values in parentheses (left margin) indicate the number of GM6-decreased genes in each category (right margin: exemplar genes most strongly repressed by GM6).

Fig. 12 shows Gene ontology biological process (BP) terms associated with PD- associated genes altered by GM6 (SBH, RNA-seq). (A) Gene ontology BP terms enriched with respect to GM6-increased PD-associated genes (FDR < 0.10; pooled from the 2 SBH RNA-seq experiments). Values in parentheses (left margin) indicate the number of GM6-increased genes in each category (right margin: exemplar genes most strongly induced by GM6). (B) Gene ontology BP terms enriched with respect to GM6-decreased PD-associated genes (FDR < 0.10; pooled from the 2 SBH RNA-seq experiments). Values in parentheses (left margin) indicate the number of GM6-decreased genes in each category (right margin: exemplar genes most strongly repressed by GM6)

Fig. 13 shows KEGG pathways in PD diagram (hsa05012; SH-5YSY cells, UM RNA-seq, 6 hours GM6 treatment). Dark grey elements are associated with genes up-regulated in GM6-treated cells compared to CTL cells, and light grey elements are associated with genes down-regulated in GM6-treated cells compared to CTL cells (see color scale, lower right). White elements have not been assigned to specific genes.. An interactive version of this diagram is available online:

htt p://^'www. genome, j p/kegg- bin/show_., pathway ?hsa05012

Figure 14 shows KEGG pathways in PD diagram (hsa05012; SH-5YSY cells, UM RNA-seq, 24 hours GM6 treatment). Dark grey elements are associated with genes up- regulated in GM6-treated cells compared to CTL cells, and light grey elements are associated with genes down-regulated in GM6-treated cells compared to CTL cells (see color scale, lower right). White elements have not been assigned to specific genes. An interactive version of this diagram is available online:

htt ://^'www. genome, j p/kegg- bin/show_.. athway ?hsa05012

Fig. 15 shows KEGG pathways in PD diagram (hsa05012; SH-5YSY cells, UM RNA-seq, 48 hours GM6 treatment). Dark grey elements are associated with genes up-regulated in GM6-treated cells compared to CTL cells, and light grey elements are associated with genes down-regulated in GM6-treated cells compared to CTL cells (see color scale, lower right). White elements have not been assigned to specific genes. An interactive version of this diagram is available online:

http://www.genome.jp/kegg-bin/show pathway'?hsa05012

Fig. 16 shows KEGG pathways in PD diagram (hsa05012; SH-5YSY cells, SBH

RNA-seq, 6 hours GM6 treatment). Dark grey elements are associated with genes up-regulated in GM6-treated cells compared to CTL cells, and light grey elements are associated with genes down-regulated in GM6-treated cells compared to CTL cells (see color scale, lower right). White elements have not been assigned to specific genes. An interactive version of this diagram is available online:

http://wwvv.genome.jp kegg-biii show pathway?hsa05()12

Fig. 17 shows KEGG pathways in PD diagram (hsa05012; SH-5YSY cells, SBH

RNA-seq, 24 hours GM6 treatment). Dark grey elements are associated with genes up-regulated in GM6-treated cells compared to CTL cells, and light grey elements are associated with genes down-regulated in GM6-treated cells compared to CTL cells (see color scale, lower right). White elements have not been assigned to specific genes.An interactive version of this diagram is available online:

http://wwvv.genome.jp kegg-biii show pathway?hsa05()12

Fig. 18. Hypothesized mechanisms of action. This analysis identified PD- associated genes regulated by GM6 consistent with 6 potential mechanisms of action. The figure summarizes these mechanisms and lists GM6-regulated genes that may play a mediating role. All genes shown in the figure were regulated by GM6 in either or both RNA-seq experiments.

DETAILED DESCRIPTION

Parkinson's is a debilitating disease for which existing drugs primarily control symptoms without preventing underlying neurodegeneration. The purpose of this particular study was to evaluate GM6 as a PD drug candidate, utilizing both in vitro and in vivo approaches to model PD pathogenesis. Additionally, to gain insight into potential mechanisms of action, we used gene expression profiling (DNA microarrays and RN A- sequencing) to identify and characterize PD-associated genes altered by GM6 treatment.

Salsolinol (SAL) is a neurotoxin byproduct of dopamine metabolism elevated in CSF from PD patients. We evaluated viability of dopaminergic human neuroblastoma SH-SY5Y cells 24 hours after treatment with GM6, SAL (100 μΜ), or the combination GM6+SAL (Figure

IA) . SAL significantly decreased cell survival by -60% (P < 0.05). This effect was abrogated, however, in cells treated concurrently with GM6 (Figure 1A). Concurrent GM6 treatment led to dose-dependent rescue of cell survival, with complete recovery of cell viability at the highest GM6 dose (10 mg/ml) (Figure 1A). Consistent with these results, GM6 also partially rescued survival of rat cortical neuronal cells treated with post-mortem CSF from PD patients (Figure

IB) . We next used DNA microarrays to identify genes altered in SH-SY5Y cells treated for 48 hrs. with GM6 (n = 2 per group, GM6 or vehicle). We identified > 1000 genes altered by GM6 (FDR < 0.10; 2-fold expression change), but focused on 104 genes associated with the PD pathway as annotated by the Kyoto Encyclopedia of Genes and Genomes (KEGG) database (Figure 2). Strikingly, nearly 80% of the KEGG PD pathway genes were decreased by GM6 (FC < 1). This down-regulation of PD-associated genes was statistically significant based upon simulation-based comparisons to randomly sampled sets of SH-SY5Y-expressed genes (Figure 3A and 3B; P < 0.001).

Overall, 61 of 104 PD-associated genes were significantly repressed by GM6 (FDR < 0.10; FC < 1.0; Figure 2). These genes were largely associated with the mitochondrial protein complex and respiratory chain (Figure 3C) and included genes associated with each mitochondrial complex (Figure 4). GM6 up-regulated 3 genes 2 fold (FDR <0.10 and FC >2.0) and 7 genes 1.5 fold (FDR < 0.10 and FC > 1.50). GM6 down-regulated 46 genes 2 fold (FDR < 0.10 and FC < 0.50) and 52 genes 1.5 fold (FDR < 0.10 and FC < 0.67}.

To assess whether GM6 could prevent dopaminergic neuron loss in vivo, we utilized 2 PD mouse models (Figures 5 and 6). First, intracerebral 6-hydroxy dopamine (6- OHDA) injection was used to damage nigrostriatal dopaminergic neurons in C57BL/6 mice (Simola et al. 2007, Neurotox Res 11:151-67). 6-OHDA-treated mice were then provided varying doses of GM6 for 5 consecutive days post-treatment (0, 1, 5, 10 or 20 mg/kg). GM6 at the highest dose almost completely abrogated 6-OHDA effects, leading to improved motor performance (Figures 5A and 5B), increased numbers of TH-positive neurons in the substantia nigra (Figure 5F), and increased brain levels of dopamine, 3,4-Dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) (Figures 5C - 5E). The experiment was repeated using an alternative PD model, i.e., mice injected with injected with l-methyl-4-phenyl-l,2,3,6- tetrahydropyridine (MPTP; Meredith and Rademacher 2011, J Parkinsons Dis 1: 19-33.) (Figure 6). Results mirrored those from the 6-OHDA experiment, with dose-dependent improvement in motor performance and dopaminergic neuron survival in GM6-treated mice (Figure 6).

We additionally report blood biomarker findings from a pilot double-blinded, randomized, placebo-controlled trial enrolling a small number of PD patients (n = 6). We have completed a pilot phase 2A placebo-controlled study to evaluate GM6 safety and potential evidence of benefit (n = 6 patients). GM6 was well-tolerated and we observed favorable changes in blood biomarkers (e.g., blood-derived neurotrophic factor, BDNF; Figure 7). Long-term data based upon larger patient cohorts appear justified and are needed to fully address clinical efficacy of GM6 in this context. GM6 is not a cocktail of different molecules. It is an endogenous embryonic stage tyrosine kinase motoronotropic factor regulator. It binds to insulin receptor, IGF1 and IGF2 receptors of the human nervous system. Our data suggest that the mechanism of action for GM6 appears to involve a number of different pathways and that, collectively, the combined action serves to promote homeostasis and correction of multiple disease states. Our data also suggest that GM6 does not act solely as an agonist or antagonist. Our findings indicate that GM6 can up and down regulate the same genes depending on whether the disease gene is below or above the normal range respectively. The findings also suggest that GM6 regulates through multiple signaling pathways in response to distress signals.

GM6 regulates 1,259 (2 fold) and 2,207 (1.5 fold) genes in the DNA microarray, within which GM6 regulates 89 (2 fold) to 238 (1.5 fold) ALS associated genes, 48 (2 fold) to 68 (1.5 fold) AD associated genes, 46 (2 fold) and to 59 (1.5 fold) PD associated genes.

We here demonstrate protective effects of GM6 against salsolinol, 6-OHDA, MPTP, and post-mortem CSF from PD patients (Figures 1, 5 and 6). It is notable that these treatments elicit neuron damage by generating reactive oxygen species (salsolinol, 6-OHDA) or by inhibition of mitochondria complex I (MPTP). Consistent with this, our microarray data show that GM6 significantly down-regulates expression of > 40 mitochondrial genes associated with PD (Figures 2 - 4). This may reflect decreased mitochondrial abundance or activity in GM6- treated cells. Potentially, therefore, protective effects of GM6 against post-mortem PD CSF, salsolinol, 6-OHDA and MPTP may involve attenuation of mitochondrial dysfunction, e.g., decreased ROS production and/or dampening of intrinsic apoptosis cascades.

These findings suggest that GM6 has the utility of therapeutic effect to treat or prevent PD disease. By preventing death of dopaminergic neurons, GM6 may improve symptoms but also address the underlying disease etiology.

Pharmaceutical Compositions

The pharmaceutical formulations provided herein may further include, as optional ingredients, pharmaceutically acceptable carriers, diluents, solubilizing or emulsifying agents, and salts of the type that are available in the art. Examples of such substances include normal saline solutions such as physiologically buffered saline solutions and water. Specific non- limiting examples of the carriers and/or diluents that are useful in the pharmaceutical formulations of the present invention include water and physiologically acceptable buffered saline solutions such as phosphate buffered saline solutions pH 7.0-8.0. Suitable pharmaceutical carriers include, but are not limited to sterile water, salt solutions (such as Ringer's solution), alcohols, polyethylene glycols, gelatin, carbohydrates such as lactose, amylose or starch, magnesium stearate, talc, silicic acid, viscous paraffin, fatty acid esters, hydroxymethylcellulose, polyvinyl pyrolidone, etc. The pharmaceutical preparations can be sterilized and desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like which do not deleteriously react with the active compounds. They can also be combined where desired with other active substances, e.g., enzyme inhibitors, to reduce metabolic degradation.

Compounds provided herein may be formulated in a pharmaceutical composition, which may include pharmaceutically acceptable carriers, thickeners, diluents, buffers, preservatives, surface active agents, neutral or cationic lipids, lipid complexes, liposomes, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients and the like in addition to the compound.

Pharmaceutical compositions may also include one or more active ingredients such as interferons, antimicrobial agents, anti-inflammatory agents, anesthetics, and the like. Formulations for parenteral administration may include sterile aqueous solutions which may also contain buffers, liposomes, diluents and other suitable additives. Pharmaceutical compositions comprising the compounds provided herein may include penetration enhancers in order to enhance the alimentary delivery of the compounds. Penetration enhancers may be classified as belonging to one of five broad categories, i.e., fatty acids, bile salts, chelating agents, surfactants and non-surfactants (Lee et al, Critical Reviews in Therapeutic Drug Carrier Systems 8, 91-192 (1991); Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems 7, 1-33 (1990)). One or more penetration enhancers from one or more of these broad categories may be included.

In certain embodiments, the pharmaceutical composition of the invention, can be administered locally, nasally, orally, gastrointestinally, intrabronchially, intravesically, intravaginally, into the uterus, sub-cutaneously, intramuscularly, periarticularly, intraarticularly, into the cerebrospinal fluid (ICSF), into the brain tissue (e.g. intracranial administration), into the spinal medulla, into wounds, intraperitoneally or intrapleurally, or systemically, e.g. intravenously, intraarterially, intraportally or into the organ directly, such as the heart.

The compounds provided herein may be administered parentally. It is sometimes preferred that certain compounds are combined with a pharmaceutically acceptable carrier or diluent to produce a pharmaceutical composition. Suitable carriers and diluents include isotonic saline solutions, for example phosphate-buffered saline. The composition may be formulated for parenteral, intramuscular, intracerebral, intravenous, subcutaneous, or transdermal administration. Uptake of nucleic acids by mammalian cells is enhanced by several known transfection techniques, for example, those that use transfection agents. The formulation which is administered may contain such agents. Example of these agents include cationic agents (for example calcium phosphate and DEAE-dextran) and lipofectants (for example lipofectamTM and transfectam TM).

Formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable. Coated gloves, condoms, and the like may also be useful. Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets or tablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders may be desirable. Compositions for parenteral administration may include sterile aqueous solutions which may also contain buffers, diluents and other suitable additives. In some cases it may be more effective to treat a patient with an compound in conjunction with other traditional therapeutic modalities in order to increase the efficacy of a treatment regimen. As used herein, the term "treatment regimen" is meant to encompass therapeutic, palliative and prophylactic modalities.

Dosing can be dependent on a number of factors, including severity and responsiveness of the disease state to be treated, and with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved. Toxicity and therapeutic efficacy of compounds provided herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. For example, for determining The LD5₀ (the dose lethal to 50% of the population) and the ED5₀ (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD5₀ /ED5₀. Compounds which exhibit large therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissues in order to minimize potential damage to uninfected cells and, thereby, reduce side effects. The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED5₀ with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC₅₀ (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography. Dosing schedules can be calculated from measurements of drug accumulation in the body of the patient.

Suitable dosage amounts may, for example, vary from about 0.1 ug up to a total dose of about 1 gram, depending upon the route of administration. Guidance as to particular dosages and methods of delivery is provided in the literature and generally available to practitioners in the art. Those skilled in the art will employ different formulations for nucleotides than for proteins or their inhibitors. Similarly, delivery of compounds provided herein will be specific to particular cells, conditions, and locations. In general, dosage is from 0.01 mg/kg to 100 mg per kg of body weight, and may be given once or more daily, weekly, monthly or yearly, or even less frequently. In the treatment or prevention of certain conditions, an appropriate dosage level will generally be about 0.001 to 100 mg per kg patient body weight per day which can be administered in single or multiple doses. Compounds according to the invention (e.g. antibodies and binding fragments thereof) may be formulated into pharmaceutical compositions for administration according to well known methodologies. Pharmaceutical compositions may, for example, comprise one or more recombinant expression constructs, and/or expression products of such constructs, in combination with a pharmaceutically acceptable carrier, excipient or diluent. Such carriers will be nontoxic to recipients at the dosages and concentrations employed. A suitable dosage may be from about 0.01 g/kg to about 1 g/kg body weight, typically by the intradermal, subcutaneous, intramuscular or intravenous route, or by other routes. A more typical dosage is about 1 g/kg to about 500 mg/kg, with about 10 μg kg, 100 μg/kg, 1 mg/kg, 10 mg/kg, 20 mg/kg, 50 mg/kg, 100 mg/kg, 200 mg/kg, 300 mg/kg, 400 mg/kg, and various ranges within these amount being still more typical for administration. It will be evident to those skilled in the art that the number and frequency of administration will be dependent upon the response of the host. "Pharmaceutically acceptable carriers" for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remingtons Pharmaceutical Sciences, Mack Publishing Co. (A.R. Gennaro edit. 1985). For example, sterile saline and phosphate-buffered saline at physiological pH may be used. Preservatives, stabilizers, dyes and even flavoring agents may be provided in the pharmaceutical composition. For example, sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid may be added as preservatives. Id. at 1449. In addition, antioxidants and suspending agents may be used. Id.

"Pharmaceutically acceptable salt" refers to salts of the compounds of the present invention derived from the combination of such compounds and an organic or inorganic acid (acid addition salts) or an organic or inorganic base (base addition salts). The compounds of the present invention may be used in either the free base or salt forms, with both forms being considered as being within the scope of the present invention.

However, pharmaceutical compositions provided herein may be in any form which allows for the composition to be administered to a patient. For example, the composition may be in the form of a solid, liquid or gas (aerosol). Typical routes of administration include, without limitation, oral, topical, parenteral (e.g., sublingually or buccally), sublingual, rectal, vaginal, and intranasal. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal, intracavernous, intrathecal, intrameatal, intraurethral injection or infusion techniques. The pharmaceutical composition is formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient. Compositions that will be administered to a patient take the form of one or more dosage units, where for example, a tablet may be a single dosage unit, and a container of one or more compounds of the invention in aerosol form may hold a plurality of dosage units.

For oral administration, an excipient and/or binder may be present. Examples are sucrose, kaolin, glycerin, starch dextrins, sodium alginate, carboxymethylcellulose and ethyl cellulose. Coloring and/or flavoring agents may be present. A coating shell may be employed.

The composition may be in the form of a liquid, e.g., an elixir, syrup, solution, emulsion or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, preferred compositions contain, in addition to one or more binding domain-immunoglobulin fusion construct or expressed product, one or more of a sweetening agent, preservatives, dye/colorant and flavor enhancer. In a composition intended to be administered by injection, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer and isotonic agent may be included.

A liquid pharmaceutical composition as used herein, whether in the form of a solution, suspension or other like form, may include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono or digylcerides which may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. Physiological saline is a preferred adjuvant. An injectable pharmaceutical composition is preferably sterile.

Compounds described herein can be used in diagnostics, therapeutics, prophylaxis, and as research reagents and in kits. Provision of means for detecting compounds of the invention can routinely be accomplished. Such provision may include enzyme conjugation, radiolabelling or any other suitable detection systems. Kits for detecting the presence or absence of compounds of the invention may also be prepared.

The compounds of the invention may also be used for research purposes. Thus, the specific activities or modalities exhibited by the compounds may be used for assays, purifications, cellular product preparations and in other methodologies which may be appreciated by persons of ordinary skill in the art.

Various aspects of the invention will now be described with reference to the following experimental section which will be understood to be provided by way of illustration only and not to constitute a limitation on the scope of the invention.

The following Examples are included for illustration and not limitation.

Example 1 - DNA microarray and RNA-Seq Analysis of GM6 regulated PD- associated genes The purpose of this study was to provide a bioinformatic analysis to assess whether GM6 alters the expression of genes associated with PD. Gene expression analyses are performed using several gene expression profiling datasets generated using DNA microarray or RNA-seq technology. These datasets evaluated effects of GM6 on gene expression in SH-SY5Y neuroblastoma cells. Effects of GM6 in SH-SY5Y neuroblastoma cells were of main interest as an in vitro dopaminergic neuron model (PMID: 20497720). Goals were to determine whether PD-associated genes exhibit unique responses to GM6 treatment, which would suggest that GM6 is impacting signaling pathways linked to core processes that underlie PD onset and/or progression. Methods

Analyses were replicated with respect to 6 datasets generated from in vitro studies in which cultured cells were treated with GM6 for varying lengths of time (Table 1).

Table 1. Expression profiling datasets. In all experiments, GM6 was applied to cells at a concentration of 1 mg/ml in type I water.

Experiments performed with n = 2 technical replicates per treatment.

bExperiments performed with n = 3 - 5 biological replicates per treatment.

Sequencing performed by University of Michigan core sequencing facility.

Sequencing performed by Phalanx Biotech.

Dataset (1) was generated using DNA microarray technology and 2 technical replicates. For these data, differential expression analyses are performed based upon estimated fold-change (FC) values and FDR estimates. Since technical and not biological replicates were used in these experiments, FDR estimates are used for heuristic purposes but are not true FDR estimates (which would require biological replication).

Datasets (2) - (6) were generated using RNA-seq technology with experiments carried out by laboratories at Sunny Biodiscovery (Santa Paula, CA) or SBH Sciences (Natick,

MA). For datasets (2) - (4), RNA-seq data were generated by core facilities at the University of Michigan (Ann Arbor, MI), while for datasets (5) and (6) RNA-seq was carried out by Phalanx

Biotech (San Diego, CA).

Genes were linked to PD based upon an association from one or more of 9 database sources (Table 2). The databases used vary in terms of their criteria and stringency for identifying a gene as PD-associated. All analyses included in this report were performed using genes linked to PD based upon at least 2 of the 9 database sources, with many highlighted genes linked to PD based upon several sources (Table 2). Table 2. Gene databases used to identify PD-associated genes. The current bioinformatic analyses focus on those genes identified with respect to at least 2+ database sources.

aDatabase of genome-wide association studies (GWAS) reported since 2008 (www.ebi.ac.uk/gwas/).

bDatabase of Medical Subject Headings (MeSH) terms based upon annotations of PubMed documents. PD-associated genes were identified based upon the MeSH term D000544 (https://www.ncbi.nlm.nih.gov/mesh).

cDisease Ontology is a disease-centered database with genes organized according to disease etiology (http://www.disease-ontology.org). PD-associated genes were identified based upon the DO identifier 10652.

dDisGeNET provides a comprehensive catalogue of genes and variants associated to human diseases (http://www.disgenet.org).

eKyoto Encyclopedia of Genes and Genomes (KEGG). PD-associated genes were identified based upon the KEGG pathway identifier hsa05010 (http://www.kegg.jp/). ^fDatabase of Disease-Gene Associations (eDGAR) (edgar.biocomp.unibo.it). The eDGAR database integrates gene-disease associations based upon the OMIM, HUMSAVAR and CLINVAR databases.

integrated compendium of annotated diseases mined from 68 data sources (http://www.malacards.org/).

hGenes were obtained from Table S6 from Mariani et al. 2016, PLoS ONE l l:e0161567. The table lists genes identified as differentially expressed between PD patient substantia nigra samples and those from healthy control subjects (whole tissue or laser microdissected samples).

'PD Gene Database. A comprehensive collection of published genetic association studies assessing PD risk, (http://www.pdgene.org/).

^JThe 2 genes linked to PD based upon 8 of 9 sources were leucine rich repeat kinase 2 (LRRK2) and synuclein alpha (SNCA).

Results

Genes associated with PD based upon 3 or more sources were more likely than randomly sampled genes to be increased by 24-48 hours of GM6 treatment (Figures 8C, 8D and 8F). For example, following 24 hours of GM6 treatment, 99 genes associated with PD based upon 3+ database sources were increased by 6% on average by GM6, which was a significantly stronger average increase than observed in randomly sampled sets of 99 genes (P = 0.006; Figure 8C). These results demonstrate a trend in which PD-associated genes tend to be increased by extended (24 - 48 hours) GM6 treatment in SH-SY5Y cells.

A second approach was to evaluate whether PD-associated genes were more responsive to GM6 stimulation compared to other genes, regardless of whether expression was elevated or repressed. The above analyses were thus repeated with average absolute fold-change [abs(log2(GM6/CTL))] as the test statistic (Figures 9). This alternative approach demonstrated similar but stronger trends, with extended (24 - 48 hour) GM6 treatment leading to stronger overall gene expression changes among genes linked to PD based upon 3 or more database sources (Figures 9C, 9D and 9F). For example, GM6 altered the expression of 99 PD-associated genes (3+ sources) by 17% on average, which was a significantly stronger average effect than observed in randomly sampled sets of 99 genes (P <0.001; Figure 9C). These results demonstrate that 24 to 48 hours of GM6 treatment in SH-SY5Y cells leads to stronger expression changes in PD-associated genes (increased or decreased expression) than observed in other genes unrelated to PD. Gene set analyses described above evaluate the average effects of GM6 on PD- associated genes (i.e., gene set analyses; Figures 8 - 9). Table 3 summarizes p-values generated from these analyses and the two alternative analysis methods.

Table 3. Gene set analysis p-value summary. The table lists p-values from simulation analyses evaluating whether average fold-change estimates (GM6/CTL) of PD- associated genes differ significantly from randomly sampled gene sets of the same size (Figures 8 -9)

aDirectional test: Test evaluates whether PD-associated genes are more strongly increased or decreased in comparison to randomly sampled gene sets. The test statistic is average fold-change (GM6/CTL).

bNon-directional test: Test evaluates whether PD-associated genes are more strongly altered (either direction) in comparison to randomly sampled gene sets. The test statistic is average absolute fold-change [abs(log2(GM6/CTL))]. It is instructive to examine trends among individual genes for which links to PD are most robustly supported by strong evidence. Overall, 3358 genes were identified as linked to PD based upon at least one database source, but only a small fraction of these were linked to PD based upon multiple sources (Data not shown). No genes were linked to PD based upon all 9 database sources, although two genes were linked to PD based upon 8 of the 9 sources (leucine rich repeat kinase 2, LRRK2; synuclein alpha, SNCA). Of these, SNCA expression was not significantly altered by GM6 in any of the 6 experimental studies (P > 0.075). However, expression of LRRK2 was decreased by GM6 in all 6 experimental studies (FC < 1.00), with a significant decrease observed in 3 of 6 experiments (P < 0.05; Data not shown). In particular, expression of LRRK2 was decreased in both RNA-seq experiments as an early (6 hour) GM6 response, with expression decreased by 34% at the 6 hour time point in the UM RNA-seq study (P = 0.0099; Data not shown), and by 23% at the 24 hour time point in the SBH RNA-seq (P=0.00683).

Likewise there was 1 gene associated with PD based upon 7 of 9 database sources (microtubule associated protein tau, MAPT), which was significantly altered by GM6 in 5 of 6 experiments. GM6 decreased expression of MAPT in the SH-SY5Y microarray dataset (FC = 0.71; P = 0.00721; Data not shown). In contrast, however, GM6 increased expression of MAPT significantly in 4 of the 5 RNA-seq-based comparisons (Data not shown). GM6 thus had significant effects on MAPT expression, although the direction of effect was not consistent between microarray and RNA-seq studies.

Among other top-ranking genes (6+ sources), it is notable that GM6 increased expression of synuclein alpha interacting protein (SNCAIP) with respect to 3 of 6 experiments, including both microarray and RNA-seq studies (Data not shown). Following 48 hours of GM6 treatment, microarray showed 58% increase (p=0.00129). Following 6 hours of GM6 treatment, RNA-seq UM showed expression of SNCAIP was increased by 62% (P = 0.00056; UM RNA- seq study). Following 24 hours of GM6 treatment, expression of SNCAIP was increased by 31% (P = 0.0488; UM RNA-seq study). Among genes linked to PD based upon 2 or more database sources, the most strongly increased in the microarray dataset included IREB2, GNAI1 and IDE, while the most strongly decreased included NDUFB1, COX6C and NDUFB6 (FDR < 0.10; Figure 2). RNA-seq analyses demonstrated strongly increased expression of SNCAIP, ACHE, GAD1 and APOE, and strongly decreased expression of SEMA5A, NEDD9 and COX4I2 (Figure 10).

The UM and SBH RNA-seq datasets were the best replicated (n = 3 - 5 per treatment) and thus further analyses were performed to characterize GM6-regulated genes identified from these experiments (Figures 11 and 12). With regards to the UM RNA-seq dataset (Figures 10A, 10B, IOC), from among all 3time points evaluated, a total of 27 GM6-increased and 19 GM6-decreased PD-associated genes were identified (FDR < 0.10).

UM, GM6-increased (27 genes) are:

MAPT; SLC41A1; RAB29; PERI; SNCAIP; HSPB1; SLC45A3; HIP1R; NEDD9; RET;

GPR37; CAPN1; ACHE; CNKSR3; LINGOl; DNM1; NPTX2; HMOX1; MAOA; DRD2; SREBF1; HGF; GAD1; GFRA1; SEPT4; BCKDK; APOE.

UM, GM6-decreased (19 genes) are:

NSF; BRINP1; NDUFS2; WNT3; ASCL1; SEMA5A; LMNB1; PRDM15; SFXN2; CD200; GRK5; MRPL3; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9; COX4I2

The 27 GM6-increased genes were significantly associated with neurotransmitter biosynthesis, negative regulation of cellular response to oxidative stress, secretion, and positive regulation of angiogenesis (Figure 11A), while the 19 GM6-decreased PD-associated genes were significantly associated with regulation of axon guidance, neuron projection extension and positive regulation of nervous system development (Figure 1 IB).

Likewise, with respect to the SBH RNA-seq dataset (Figures 10D, 10E), from the 2 time points evaluated, a total of 43 GM6-increased and 26 GM6-decreased PD-associated genes were identified (FDR < 0.10).

SBH, GM6-increased (43 genes) are:

CAPN1; HGF; GST02; SNCB; ACHE; BCKDK; HSPB1; DNM1; WNT3; VEGFA; RAB29; HIP1R; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; SLC45A3; KRTCAP2; RET; SLC50A1; LINGOl; C8orf4; PERI; MAPT; CRHR1-IT1; VDAC1; NOS1; NPTX2; BST1; CRYAB; SOD2; HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl; TMEM229B; TGM2; HSPA1A. SBH, GM6-decreased (26 genes) are:

ATXN3; MCCC1; NDUFB2; HSPA8; LMNB1; MRPL3; SIPA1L2; CAT; TP53; PRDM15; CNNM2; SLC25A4; NDUFS2; PRDX3; BORCS7; ZNF646; TRAF6; SIAH1; HTT; DDRGK1; UNC13B; GIGYF2; P2RX7; CD200; NDUFS4; LRRK2.

The 43 GM6-increased PD-associated genes were associated with regulation of neurogenesis, signaling, apoptotic process and reactive oxygen species metabolism (Figure 12A), and the 26 GM6-decreased genes were associated with the metabolism of ribonucleoside triphosphate, purine nucleoside triphosphate, glycosyl metabolism, purine nucleoside monophosphate and reactive oxygen species (Figure 12B).

There was a total of 56 PD-associated gene increased by GM6 in RNA-seq:

MAPT, RAB29, PERI, HSPB1, SLC45A3, HIP1R, RET, CAPN1, ACHE, LINGOl, DNM1, NPTX2, HGF, BCKDK, SLC41A1, SNCAIP, NEDD9, GPR37, CNKSR3, HMOX1, MAOA, DRD2, SREBF1, GAD1, GFRA1, SEPT4, APOE, GST02, SNCB, WNT3, VEGFA, SYT4, RTN3, KCNN3, PRSS53, UBE2L6, KRTCAP2, SLC50A1, C8orf4, CRHR1-IT1 VDAC1, NOS1, BST1, CRYAB, SOD2, HIVEP3, ARNTL, S100B, LY6E, CTSB, IGF2, DDIT4, NQOl, TMEM229B, TGM2, HSPA1AGM6- increased PD-associated genes from RNA-seq with Fold Change greater than 1.5 (FC>1.5) are SNCAIP, ACHE, GAD1, HMOX1, LINGOl, GFRAl, MAOA, RAB29, S100B, IGF2,TGM2.

There was a total of 40 PD-associated gene decreased by GM6 in RNA-seq: NDUFS2, LMNB1, PRDM15, CD200, MRPL3, NSF, BRINP1, WNT3, ASCL1, SEMA5A, SFXN2, GRK5, CXCR4, DDIT4, GSTA4, DDC, VEGFA, NEDD9, COX4I2,

ATXN3, MCCC1, NDUFB2, HSPA8, SIPA1L2, CAT, TP53, CNNM2, SLC25A4, PRDX3, BORCS7, ZNF646, TRAF6, SIAH1, HTT, DDRGK1, UNC13B, GIGYF2, P2RX7, NDUFS4, LRRK2. GM6-decreased PD-associated genes from RNA-seq with FC<0.67 are PRDM15, SEMA5A, VEGFA, NEDD9, and COX4I2

Patterns of GM6 gene regulation were visualized using color-coded KEGG PD pathway diagrams (hsa05012). Dark grey elements are associated with genes up-regulated in GM6-treated cells compared to CTL cells, and light grey elements are associated with genes down-regulated in GM6-treated cells compared to CTL cells (see color scale, lower right). White elements have not been assigned to specific genes. This diagram provides a gene mapping to pathways leading to cell death and the production of Lewy bodies and disease-related peptides in dopaminergic neurons, including key processes related to disease-mediating ubiquitin and mitochondrial pathways (Figure 4, Figures 13 - 17). Pathway maps showed that GM6 induced down-regulation of ubiquitin and mitochondrial pathway components in the microarray dataset, including genes associated with protein kinase A (PKA) associated with motor impairment, as well as genes linked to ubiquitin B, tyrosine 3-monooxygenase and mitochondrial enzymes (Figure 13). Some of these patterns were attenuated with respect to RNA-seq comparisons (Figures 11 - 17). However, consistent with the microarray dataset, GM6 decreased expression of genes encoding tyrosine 3-monooxygenase as well as mitochondrial enzyme components, such as NADH dehydrogenase (ubiquinone) 1 subunit C2, ubiquinol-cytochrome c reductase subunit 10, cytochrome c oxidase subunit 6b, and F-type H+-transporting ATPase subunit d (UM-RNA seq experiments; 24 - 48 hours GM6 treatment; Figures 14 and 15).

The purpose of this study was to identify PD-associated genes regulated by GM6 and to characterize functional properties of such genes. There was evidence that PD-associated genes are more likely to be altered by GM6 in comparison to other human genes (Figures 9C, 9D and 9F). The analysis further identified GM6-regulated genes with potentially important roles in PD development and progression (Figure 18). These genes were categorized with respect to 6 hypothesized mechanisms of action, including (1) attenutation of kinase dysfunction via LRKK2 down-regulation, (2) anti-cholinergic actions, (3) pro-GABAergic effects, (4) enhancement of GDNF activity through activation of a GRFA1-RET axis, (5) improved oxidative stress defenses and (6) blunting of free radical generation through decreased mitochondrial activity or abundance (Figure 18). Collectively, these effects are hypothesized to broadly impact neurotransmitter balance in PD, by improving survival of dopaminergic neurons to promote dopamine availability (mechanisms 1, 4, 5 and 6) but also by curbing cholinergic activity with concomitant enhancement of GABAergic signaling (mechanisms 2 and 3).

Claims

What is claimed is:

1. A method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide with the peptide sequence FSRYAR (GM6) to a subject to regulate the pathology of Parkinson' s disease, wherein GM6 regulates more than one downstream target gene to regulate the pathology of the disease.

2. A method according to claim 1, wherein GM6 regulates multiple targets to regulate the pathology of Parkinson' s disease.

3. A method according to claim 1, wherein GM6 acts as a master regulator of multiple targets to regulate the pathology of Parkinson's disease to treat or prevent Parkinson's disease.

4. A method according to claim 1, wherein GM6 functions both as an agonist and an antagonist for different targets that regulate the pathology of Parkinson's disease.

5. A method according to claim 1, wherein GM6 functions both as an agonist and an antagonist for different targets that regulate the pathology of Parkinson's disease, wherein GM6 regulates the pathology of Parkinson's disease by up regulating some targets and downregulating other targets to achieve a homeostasis.

6. A method of treating or preventing Parkinson' s disease, the method comprising the administering a MNTF peptide consisting of the amino acids FSRYAR (GM6) to a subject to regulate the pathology of Parkinson' s disease, wherein GM6 regulates more than one downstream target to inhibit mitochondria-mediated neuronal cell death associated with

Parkinson's disease.

7. A method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide with the peptide sequence FSRYAR (GM6) to a subject, wherein GM6 regulates more than one of the following downstream targets to regulate the pathology of the disease: GNAI1, CASP3, PRKACB, UBE2L3, CASP9, UBE2J1, SNCAIP, ATP5B, GPR37, GNAI3, SLC18A1, UBE2G1, APAF1, UQCRC2, NDUFS1, NDUFV3, PINK1, PARK2, SNCA, NDUFA4L2, NDUFV1, ATP5E, UBE2J2, NDUFS2, UQCRC1, HTRA2, ADORA2A, COX6A2, COX7A1, LRRK2, PPIF, SLC18A2, VDAC3, SDHD, SLC25A6, COX4I2,

NDUFC2, SLC25A31, GNAL, PRKACG, COX7B2, NDUFA10, SLC6A3, UQCRB, ATP5J, COX6B2, DRD2, ADCY5, UBE2L6, GNAI2, ATP5F1, UCHL1, ATP5C1, COX7A2L, UBB, SLC25A5, TH, NDUFB5, NDUFA5, UBE2G2, SDHB, CYCS, CYC1, COX4I1, NDUFB3, ATP5G3, COX8A, NDUFA9, PRKACA, UQCRQ, NDUFS3, COX6B1, NDUFA1, SLC25A4, NDUFS4, COX5B, NDUFA6, NDUFS8, ATP5D, NDUFA13, NDUFB9, NDUFS5, NDUFB10, NDUFBll, NDUFA7, NDUFS6, NDUFA8, NDUFA2, UQCRH, COX7C, NDUFB4, NDUFB6, NDUFC1, COX6C, NDUFAB1, NDUFB2, NDUFB7, COX7A2, COX7B, ATP50, COX5A, PARK7, NDUFA12, and NDUFBl.

8. A method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide with the peptide sequence FSRYAR (GM6) to a subject, wherein GM6 down-regulates at least one of the following downstream targets to regulate the pathology of the disease: NDUFBl, NDUFA12, PARK7, COX5A, ATP50, COX7B, COX7A2, NDUFB7, NDUFB2, NDUFAB1, COX6C, NDUFC1, NDUFB6, NDUFB4, COX7C, UQCRH, NDUFA2, NDUFA8, NDUFS6, NDUFA7, NDUFBl l, NDUFBIO, NDUFS5, NDUFB9, NDUFA13, ATP5D, NDUFS8, NDUFA6, COX5B, NDUFS4, SLC25A4, NDUFA1, COX6B1, NDUFS3, UQCRQ, PRKACA, NDUFA9, COX8A, ATP5G3, NDUFB3, COX4I1, CYC1, CYCS, SDHB, UBE2G2, NDUFA5, NDUFB5, TH, SLC25A5, UBB, COX7A2L, and UCHL1.

9. A method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide consisting of the amino acids FSRYAR (GM6) to a subject, wherein GM6 down-regulates at least one of the following downstream targets to regulate the pathology of the disease: NDUFBl, NDUFA12, PARK7, COX5A, ATP50, COX7B, COX7A2, NDUFB7, NDUFB2, NDUFAB1, COX6C, NDUFC1, NDUFB6, NDUFB4, COX7C, UQCRH, NDUFA2, NDUFA8, NDUFS6, NDUFA7, NDUFBll, NDUFBIO, NDUFS5, NDUFB9,

NDUFA13, ATP5D, NDUFS8, NDUFA6, COX5B, NDUFS4, SLC25A4, NDUFA1, COX6B1, NDUFS3, UQCRQ, PRKACA, NDUFA9, COX8A, ATP5G3, NDUFB3, COX4I1, NDUFB5, and TH.

10. A method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide consisting of the amino acids FSRYAR (GM6) to a subject, wherein GM6 up- regulates at least one of the following downstream targets to regulate the pathology of the disease: GNAI1, CASP3, PRKACB, UBE2L3, CASP9, UBE2J1, SNCAIP.

11. A method of treating or preventing Parkinson' s disease, the method comprising administering a MNTF peptide consisting of the amino acids FSRYAR (GM6) to a subject, wherein GM6 up- regulates at least one of the following downstream targets to regulate the pathology of the disease: GNAI1, CASP3, PRKACB.

12. A method of treating or preventing Parkinson's disease, the method comprising administering a MNTF peptide with the peptide sequence FSRYAR (GM6) to a subject, wherein GM6 regulates the genes in pathways selected from the following: mitochondrial protein complex, respiratory chain complex, mitochondrial respiratory chain complex, NADH dehydrogenase complex, mitochondrial envelope, envelope, organelle membrane, catalytic complex, intracellular organelle part, mitochondrion, macromolecular complex, mitochondrial inner membrane, mitochondrial membrane part, mitochondrial respiratory chain, mitochondrial intermembrane space.

13. A method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID

NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by regulating the expression of one or more genes selected from the group consisting of: MAPT; RAB29; PERI; HSPB1; SLC45A3; HIP1R; RET; CAPN1; ACHE; LINGOl; DNM1; NPTX2; HGF; BCKDK; SLC41A1; SNCAIP; NEDD9; GPR37; CNKSR3; HMOX1; MAO A; DRD2; SREBF1; GAD1; GFRA1; SEPT4; APOE; GST02; SNCB; WNT3; VEGFA; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; KRTCAP2; SLC50A1; C8orf4; CRHR1- IT1 VDAC1; NOS1; BST1; CRYAB; SOD2; HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl; TMEM229B; TGM2 and HSPA1A.

14. A method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID

NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by regulating the expression of one or more genes selected from the group consisting of: NDUFS2; LMNBl; PRDM15; CD200; MRPL3; NSF; BRINPl; WNT3; ASCLl; SEMA5A; SFXN2; GRK5; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9 and COX4I2; ATXN3; MCCC1; NDUFB2; HSPA8; SIPA1L2; CAT; TP53; CNNM2; SLC25A4; PRDX3; BORCS7; ZNF646; TRAF6; SIAH1; HTT; DDRGK1; UNC13B; GIGYF2; P2RX7; NDUFS4; and LRRK2.

15. A method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by regulating the expression of one or more genes selected from the group consisting of: MAPT; SLC41A1; RAB29; PERI; SNCAIP; HSPB1; SLC45A3; HIP1R; NEDD9; RET; GPR37; CAPN1; ACHE; CNKSR3; LINGOl; DNM1; NPTX2; HMOX1;

MAO A; DRD2; SREBF1; HGF; GAD1; GFRA1; SEPT4; BCKDK and APOE.

16. A method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by regulating the expression of one or more genes selected from the group consisting of: NSF; BRINP1; NDUFS2; WNT3; ASCL1; SEMA5A; LMNB1; PRDM15; SFXN2; CD200; GRK5; MRPL3; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9 and COX4I2.

17. A method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by regulating the expression of one or more genes selected from the group consisting of: CAPN1; HGF; GST02; SNCB; ACHE; BCKDK; HSPB1; DNM1; WNT3; VEGFA; RAB29; HIP1R; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; SLC45A3; KRTCAP2; RET; SLC50A1; LINGOl; C8orf4; PERI; MAPT; CRHR1-IT1; VDAC1; NOS1; NPTX2; BST1; CRYAB; SOD2; HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl; TMEM229B; TGM2 and HSPA1A.

18. A method of treating or preventing Parkinson's disease, the method comprising the steps of: i) administering a MNTF peptide consisting of the amino acids FSRYAR [SEQ ID

NO:2] (GM6) to a subject to inhibit or prevent Parkinson's disease associated neuron loss or dysfunction by regulating the expression of one or more genes selected from the group consisting of: ATXN3; MCCC1; NDUFB2; HSPA8; LMNB1; MRPL3; SIPA1L2; CAT; TP53; PRDM15; CNNM2; SLC25A4; NDUFS2; PRDX3; BORCS7; ZNF646; TRAF6; SIAH1; HTT; DDRGK1; UNC13B; GIGYF2; P2RX7; CD200; NDUFS4 and LRRK2.

19. A method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: MAPT; RAB29; PERI; HSPB1; SLC45A3; HIP1R; RET; CAPN1; ACHE; LINGOl; DNM1; NPTX2; HGF; BCKDK; SLC41A1; SNCAIP; NEDD9; GPR37; CNKSR3; HMOX1; MAO A; DRD2; SREBF1; GAD1; GFRA1; SEPT4; APOE; GST02; SNCB; WNT3; VEGFA; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; KRTCAP2; SLC50A1; C8orf4; CRHR1-IT1 VDAC1; NOS1; BST1; CRYAB; SOD2; HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl;

TMEM229B; TGM2 and HSPA1A in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson' s disease.

20. A method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: NDUFS2; LMNB1; PRDM15; CD200; MRPL3; NSF; BRINP1; WNT3; ASCL1; SEMA5A; SFXN2; GRK5; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9 and COX4I2; ATXN3; MCCC1; NDUFB2; HSPA8; SIPA1L2; CAT; TP53; CNNM2; SLC25A4; PRDX3; BORCS7; ZNF646; TRAF6; SIAH1; HTT; DDRGK1; UNC13B; GIGYF2; P2RX7; NDUFS4; and LRRK2 in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson' s disease.

21. A method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: MAPT; SLC41A1; RAB29; PERI; SNCAIP; HSPB1; SLC45A3; HIP1R; NEDD9; RET; GPR37;

CAPN1; ACHE; CNKSR3; LINGOl; DNM1; NPTX2; HMOX1; MAOA; DRD2; SREBF1; HGF; GAD1; GFRA1; SEPT4; BCKDK and APOE in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of

Parkinson's disease.

22. A method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: NSF;

BRINP1; NDUFS2; WNT3; ASCL1; SEMA5A; LMNB1; PRDM15; SFXN2; CD200; GRK5; MRPL3; CXCR4; DDIT4; GSTA4; DDC; VEGFA; NEDD9 and COX4I2 in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson' s disease.

23. A method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: CAPN1;

HGF; GST02; SNCB; ACHE; BCKDK; HSPB1; DNM1; WNT3; VEGFA; RAB29; HIP1R; SYT4; RTN3; KCNN3; PRSS53; UBE2L6; SLC45A3; KRTCAP2; RET; SLC50A1; LINGOl;

C8orf4; PERI; MAPT; CRHR1-IT1; VDAC1; NOS1; NPTX2; BST1; CRYAB; SOD2;

HIVEP3; ARNTL; S100B; LY6E; CTSB; IGF2; DDIT4; NQOl; TMEM229B ; TGM2 and

HSPAIA in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of Parkinson' s disease.

24. A method of diagnosing Parkinson's disease in a patient, comprising: detecting the level of expression of one or more genes selected from the group consisting of: ATXN3; MCCCl; NDUFB2; HSPA8; LMNBl; MRPL3; SIPA1L2; CAT; TP53; PRDM15; CNNM2; SLC25A4; NDUFS2; PRDX3; BORCS7; ZNF646; TRAF6; SIAHl; HTT; DDRGKl; UNC13B; GIGYF2; P2RX7; CD200; NDUFS4 and LRRK2 in a biological sample from said patient, wherein differential expression of said one or more gene variants in the sample as compared to control levels of expression of said one or more genes or gene variants is indicative of

Parkinson's disease.

25. A method according to any one of claims 1-24, wherein GM6 is used in combination with another drug or therapy.

26. A method according to claim 1, wherein administration of GM6 regulates one or more genes involved in attenuation of kinase dysfunction via LRKK2 down-regulation.

27. A method according to claim 1, wherein administration of GM6 regulates one or more genes involved in anti-cholinergic actions.

28. A method according to claim 1, wherein administration of GM6 regulates one or more genes involved in pro-GABAergic effects.

29. A method according to claim 1, wherein administration of GM6 regulates one or more genes involved in enhancement of GDNF activity through activation of a GRFAl-RET axis.

30. A method according to claim 1, wherein administration of GM6 regulates one or more genes involved in improved oxidative stress defenses.

31. A method according to claim 1, wherein administration of GM6 regulates one or more genes involved in blunting of free radical generation through decreased mitochondrial activity or abundance

32. A method according to claim 1, wherein GM6 is administered intravenously.

33. A method according to claim 1, wherein GM6 is administered subcutaneously.

34. A method according to claim 1, wherein GM6 is administered orally.

35. A method according to claim 1, wherein GM6 is administered by combination of intravenously and subcutaneously.