METHODS RELATED TO PARKINSON?S DISEASE AND SYNUCLEINOPATHIES

1 . A method of generating antibodies useful for treating Parkinson's disease (PD) and other synucleinopathies, comprising (a) immunizing a non-human animal with an immunogen composition comprising an alpha-synuclein (α-syn) derived polypeptide or a polymer exhibiting the same conformational epitope as the polypeptide, and (b) isolating one or more antibodies that specifically recognize the polypeptide; wherein the α-syn derived polypeptide comprises a conformationally distinct and nonfibrillar α-syn variant with mitotoxicity.

2 . The method of claim 1 , wherein the α-syn derived polypeptide comprises phosphorylated Ser 129 .

3 . The method of claim 1 , wherein the α-syn derived polypeptide is immunoreactive with anti-phospho-Ser129 antibody GTX50222, lot 821505177.

4 . The method of claim 1 , wherein the α-syn derived polypeptide is not immunoreactive with fibrillar Pα-synF-recognizing 81A and/or antibody MJF-R13.

5 . The method of claim 1 , wherein the α-syn derived polypeptide is an α-syn variant with a deletion of about 0 to 25 N-terminal amino acid residues and/or a deletion of about 0 to 25 C-terminal amino acid residues relative to a full length α-syn protein.

6 . The method of claim 1 , wherein the α-syn derived polypeptide is extracted from Pα-syn* inclusions present in a cell culture, brains of animal models of PD and other synucleinopathies, or brains of patients with PD and other synucleinopathies.

7 . The method of claim 1 , wherein the immunogen composition further comprises an adjuvant.

8 . The method of claim 1 , wherein the antibody is isolated by phage display.

9 . The method of claim 1 , further comprising examining the isolated antibodies for a therapeutic activity.

10 . The method of claim 9 , wherein the therapeutic activity is inhibition of a toxic activity in a cellular or organism model of synucleinopathy.

11 . The method of claim 14 , wherein the therapeutic activity is a reduction in the generation and propagation of pathogenic phosphorylated α-syn.

12 . The method of claim 1 , wherein the polypeptide is derived from a human α-syn.

13 . The method of claim 12 , wherein the human α-syn comprises at least a 50% sequence identity to SEQ ID NO:1, variants or fragments thereof.

14 . The method of claim 13 , wherein the human α-syn comprises SEQ ID NO:1, varaints or fragments thereof.

15 . The method of claim 1 , wherein the mitotoxicity is inducing mitochondrial dysfunction and structural damage resulting in mitophagy

16 . A method for identifying potential therapeutic agents for treating PD and other synucleinopathies, comprising (a) contacting with or administering to a cell or animal model of PD and other synucleinopathies a plurality of candidate agents, (b) detecting in a specific candidate agent-treated model a disruption or decreased formation of an alpha-synuclein (α-syn) derived polypeptide relative to untreated control model, thereby identifying the specific candidate agent as a potential therapeutic agent for treating PD and other synucleinopathies; wherein the α-syn derived polypeptide comprises a conformationally distinct and nonfibrillar α-syn variant with mitotoxicity.

17 . The method of claim 16 , wherein the α-syn derived polypeptide comprises phosphorylated Ser 129 .

18 . The method of claim 16 , wherein the α-syn derived polypeptide is an α-syn variant with a deletion of about 0 to 25 N-terminal amino acid residues and/or a deletion of about 0 to 25 C-terminal amino acid residues relative to a full length α-syn protein.

19 . The method of claim 16 , wherein the mitotoxicity is inducing mitochondrial dysfunction and structural damage resulting in mitophagy.

20 . The method of claim 16 , wherein the potential therapeutic agent inhibits α-syn derived polypeptide mediated formation of phosphorylated acetyl-CoA carboxylase (ACC) aggregates.

21 . The method of claim 16 , wherein the potential therapeutic agent inhibits α-syn derived polypeptide mediated phosphorylation of glycogen synthase kinase 3 beta (GSK3β) and mitogen-activated protein kinases (MAPKs) comprising mitogen-activated protein kinase kinase 4 (MKK4), c-Jun N-terminal kinase (JNK), p38 or extracellular signal-regulated kinase 5 (ERK5).

22 . The method of claim 16 , wherein the potential therapeutic agent inhibits α-syn derived polypeptide mediated formation of phosphorylated tau aggregates.

23 . The method of claim 16 , wherein the potential therapeutic agent inhibits α-syn derived polypeptide mediated synaptic toxicity and loss of dendritic spines of neurons.

24 . The method of claim 16 , wherein the α-syn derived polypeptide is immunoreactive with anti-phospho-Ser129 antibody GTX50222, lot 821505177.

25 . The method of claim 16 , wherein the α-syn derived polypeptide is not immunoreactive with fibrillar Pα-synF-recognizing 81A and/or antibody MJF-R13.

26 . The method of claim 16 , further comprising examining the potential therapeutic agent for a therapeutic activity.

27 . The method of claim 26 , wherein the therapeutic activity is inhibition of a toxic activity in a cellular or organism model of synucleinopathy.

28 . The method of claim 26 , wherein the therapeutic activity is a reduction in the generation and propagation of pathogenic phosphorylated α-syn.

29 . The method of claim 16 , wherein the polypeptide is derived from a human α-syn.

30 . The method of claim 29 , wherein the human α-syn comprises at least a 50% sequence identity to SEQ ID NO:1, variants or fragments thereof.

31 . The method of claim 30 , wherein the human α-syn comprises SEQ ID NO:1, variants or fragments thereof.

32 . A method for identifying potential therapeutic agents for treating PD and other synucleinopathies, comprising (a) contacting with or administering to a cell or animal model of PD and other synucleinopathies or a α-syn derived polypeptide extracted from Pα-syn* inclusions present in a cell culture, brains of animal models, or brains of patients with PD and other synucleinopathies. a plurality of candidate agents, (b) detecting binding of a candidate agent to a alpha-synuclein (α-syn) derived polypeptide specific to the candidate agent-treated relative to untreated control model, thereby identifying the specific candidate agent as a potential therapeutic agent for treating PD and other synucleinopathies; wherein the α-syn derived polypeptide comprises a conformationally distinct and nonfibrillar α-syn variant with mitotoxicity.

33 . The method of claim 32 , wherein the α-syn derived polypeptide comprises phosphorylated Ser 129 .

34 . The method of claim 32 , wherein the α-syn derived polypeptide is an α-syn variant with a deletion of about 0 to 25 N-terminal amino acid residues and/or a deletion of about 0 to 25 C-terminal amino acid residues relative to a full length α-syn protein.

35 . The method of claim 32 , wherein the mitotoxicity is inducing mitochondrial dysfunction and structural damage resulting in mitophagy.

36 . The method of claim 32 , wherein the potential therapeutic agent inhibits α-syn derived polypeptide mediated formation of phosphorylated acetyl-CoA carboxylase (ACC) aggregates.

37 . The method of claim 32 , wherein the potential therapeutic agent inhibits α-syn derived polypeptide mediated phosphorylation of glycogen synthase kinase 3 beta (GSK3β) and mitogen-activated protein kinases (MAPKs) such as mitogen-activated protein kinase kinase 4 (MKK4), c-Jun N-terminal kinase (JNK), p38 and extracellular signal-regulated kinase 5 (ERK5).

38 . The method of claim 32 , wherein the potential therapeutic agent inhibits α-syn derived polypeptide mediated formation of phosphorylated tau aggregates.

39 . The method of claim 32 , wherein the potential therapeutic agent inhibits α-syn derived polypeptide mediated synaptic toxicity and loss of dendritic spines of neurons.

40 . The method of claim 32 , wherein the α-syn derived polypeptide is immunoreactive with anti-phospho-Ser129 antibody GTX50222, lot 821505177.

41 . The method of claim 32 , wherein the α-syn derived polypeptide is not immunoreactive with fibrillar Pα-synF-recognizing 81A and/or antibody MJF-R13.

42 . The method of claim 32 , further comprising examining the potential therapeutic agent for a therapeutic activity.

43 . The method of claim 42 , wherein the therapeutic activity is inhibition of a toxic activity in a cellular or organism model of synucleinopathy.

44 . The method of claim 42 , wherein the therapeutic activity is a reduction in the generation and propagation of pathogenic phosphorylated α-syn.

45 . The method of claim 32 , wherein the polypeptide is derived from a human α-syn.

46 . The method of claim 32 , wherein the human α-syn comprises at least a 50% sequence identity to SEQ ID NO:1, variants or fragments thereof.

47 . The method of claim 34 , wherein the human α-syn comprises SEQ ID NO:1, variants or fragments thereof.

48 . A method of diagnosing or monitoring disease progression in patients affected by PD and other synucleinopathies, comprising detecting the presence and/or quantifying the amount of a conformationally distinct and nonfibrillar α-syn variant with mitotoxicity.

49 . The method of claim 48 , wherein the α-syn derived polypeptide comprises phosphorylated Ser 129 .

50 . The method of claim 48 , wherein the α-syn variant comprises a deletion of about 0 to 25 N-terminal amino acid residues and/or a deletion of about 0 to 25 C-terminal amino acid residues relative to a full length α-syn protein.

51 . The method of claim 48 , wherein the mitotoxicity is inducing mitochondrial dysfunction and structural damage resulting in mitophagy.

52 . The method of claim 48 , wherein the method detects α-syn variant mediated formation of phosphorylated acetyl-CoA carboxylase (ACC) aggregates.

53 . The method of claim 48 , wherein the method detects α-syn variant mediated phosphorylation of glycogen synthase kinase 3 beta (GSK3β) and mitogen-activated protein kinases (MAPKs) such as mitogen-activated protein kinase kinase 4 (MKK4), c-Jun N-terminal kinase (JNK), p38 and extracellular signal-regulated kinase 5 (ERK5).

54 . The method of claim 48 , wherein the method detects α-syn variant mediated formation of phosphorylated tau aggregates.

55 . The method of claim 48 , wherein the method detects α-syn variant mediated synaptic toxicity and loss of dendritic spines of neurons.

56 . The method of claim 48 , wherein the diagnosis or disease monitoring is performed with a tissue or body fluid sample obtained from subjects affected by PD and other synucleinopathies.

57 . An engineered cell or transgenic non-human animal comprising a transgene encoding an alpha-synuclein (α-syn) derived polypeptide, wherein the α-syn derived polypeptide consists of a deletion of about 0 to 25 N-terminal amino acid residues and a deletion of about 0 to 25 C-terminal amino acid residues of a full length α-syn protein.

58 . The engineered cell of claim 57 , which is a neuronal cell.

59 . The transgenic non-human animal of claim 57 , which is a rodent.

60 . An engineered cell or transgenic non-human animal comprising a transgene encoding an alpha-synuclein (α-syn) derived polypeptide, wherein the α-syn derived polypeptide harbours a mutation in one or several amino acid residues of a full length or truncated α-syn protein rendering the α-syn variant prone to adopt a distinct and nonfibrillar α-syn conformation with mitotoxicity.

61 . The engineered cell of claim 60 , which is a neuronal cell.

62 . The transgenic non-human animal of claim 60 , which is a rodent.

63 . A method of generating small molecules useful for treating Parkinson's disease (PD) and other synucleinopathies, comprising (a) performing structure-based drug design directed towards the conformational epitope of an immunogen composition comprising an alpha-synuclein (α-syn) derived polypeptide or a polymer exhibiting the same conformational epitope as the polypeptide, and (b) selecting a small molecule specifically recognizing the conformational epitope of an α-syn derived polypeptide; wherein the α-syn derived polypeptide comprises a conformationally distinct and nonfibrillar α-syn variant with mitotoxicity.

64 . The method of claim 63 , wherein the α-syn derived polypeptide comprises phosphorylated Ser 129 .

65 . The method of claim 63 , wherein the α-syn derived polypeptide is immunoreactive with anti-phospho-Ser129 antibody GTX50222, lot 821505177.

66 . The method of claim 63 , wherein the α-syn derived polypeptide is not immunoreactive with fibrillar Pα-synF-recognizing 81A and/or antibody MJF-R13.

67 . The method of claim 64 , wherein the α-syn derived polypeptide is an α-syn variant with a deletion of about 0 to 25 N-terminal amino acid residues and/or a deletion of about 0 to 25 C-terminal amino acid residues relative to a full length α-syn protein.

68 . The method of claim 63 , wherein the mitotoxicity is inducing mitochondrial dysfunction and structural damage resulting in mitophagy.

69 . The method of claim 63 , wherein the small molecule inhibits α-syn derived polypeptide mediated formation of phosphorylated acetyl-CoA carboxylase (ACC) aggregates.

70 . The method of claim 63 , wherein the small molecule inhibits α-syn derived polypeptide mediated phosphorylation of glycogen synthase kinase 3 beta (GSK3β) and mitogen-activated protein kinases (MAPKs) such as mitogen-activated protein kinase kinase 4 (MKK4), c-Jun N-terminal kinase (JNK), p38 and extracellular signal-regulated kinase 5 (ERK5).

71 . The method of claim 63 , wherein the small molecule inhibits α-syn derived polypeptide mediated formation of phosphorylated tau aggregates.

72 . The method of claim 63 , wherein the small molecule inhibits α-syn derived polypeptide mediated synaptic toxicity and loss of dendritic spines of neurons.

73 . The method of claim 63 , wherein the α-syn derived polypeptide is extracted from Pα-syn* inclusions present in a cell culture, brains of animal models of PD and other synucleinopathies, or brains of patients with PD and other synucleinopathies.

74 . The method of claim 63 , further comprising examining the selected small molecule for a therapeutic activity.

75 . The method of claim 74 , wherein the therapeutic activity is inhibition of a toxic activity in a cellular model of synucleinopathy or a reduction in the generation and propagation of pathogenic phosphorylated α-syn.

76 . The method of claim 63 , wherein the polypeptide is derived from a human α-syn.

77 . The method of claim 76 , wherein the human α-syn comprises at least a 50% sequence identity to SEQ ID NO:1, variants or fragments thereof.

78 . The method of claim 77 , wherein the human α-syn consists of SEQ ID NO:1.