KASPP (LRRK2) gene, its production and use for the detection and treatment of neurodegenerative disorders

#The present invention refers to a newly discovered gene named KASPP for Kinase Associated with Parkinsonism with Pleiomorphic Pathology or alternatively named LRRK2 for Leucine-Rich Repeat Kinase 2, its production, biochemical characterization and use for the detection and treatment of neurodegenerative disorders, such as Parkinson disease (PD) including, without limitation, sporadic PD, Alzheimer disease (AD), amyotrophic lateral sclerosis (ALS), and other synucleinopathies and/or tauopathy as well as several polymorphisms and mutations in the KASPP/LRRK2 gene segregated with PD.

#Parkinson's disease (PD) is the second most neurodegenerative disorder affecting 1-2% of the population aged 65 and older characterized by a progressive loss of dopaminergic neurons of the substantia nigra, associated with the formation of fibrillar aggregates composed of α-synuclein and other proteins (Lewy bodies and Lewy neurites). In most cases, PD occurs as a sporadic disease of unknown etiology, but in rare instances, point mutations or multiplications of the α-synuclein gene can cause autosomal-dominant parkinsonism which resembles the sporadic disease in many aspects. Recessive forms of parkinsonism have been recognized, which are caused by mutations in the genes for parkin (Kitada T. et al., Nature, 392, 605-608, 1998), DJ-1 (Bonifati V. et al., Science, 299, 256-259, 2002) and PINK1 (Valente E. M. et al., Science, 304 (5674), 1158-1160, 2004). Additional loci have been mapped on chromosomes 2p (Gasser T. et al., Nat. Genet., 18, 262-265, 1998), 12cen (Funayama M. et al., Ann. Neurol., 51(3), 296-301, 2002), 1q (Hicks A. A. et al., Ann. Neurol., 52(5), 549-555, 2002), and 2q (Pankratz N. et al., Am. J. Hum. Genet., 72(4), 1053-1057, 2003). In more than 10% of patients with PD one or more relatives are also affected by this disorder (Elbaz et al., Neurology, 52:1876-82, 1999). However, genetic causes are only very rarely found.

#As α-synuclein aggregation is a pathologic feature both in the common sporadic and a dominantly inherited form of PD, and also in other neurodegenerative diseases, such as dementia with Lewy bodies (DLB) and multiple systems atrophy (MSA), those diseases have collectively been called “synucleinopathies”. Other forms of parkinsonism are associated with the accumulation of filaments composed of the microtubule associated protein tau (MAPT). Mutations in this gene explain at least a subgroup of families with frontotemporal dementia with parkinsonism (FTDP-17; Ghetti B. et al., “Frontotemporal dementia and parkinsonism linked to chromosome 17 associated with tau gene mutations (FTDP-17)”. In: Dickson D. W., “Neurodegeneration: the molecular pathology of dementia and movement disorders” ISN Neuropath Press, Basal, 86-102, 2003), while sporadic cases with tau pathology most commonly present as progressive supranuclear palsy (PSP) or corticobasal degeneration (CBD). Based on the putative central role of the tau protein in these diseases, they have been called “tauopathies”.

#FIG. 1 shows the pedigree structure of the two largest families: A (German-Canadian), D (Western Nebraska) and with mutations. Blackened symbols denote affected family members; asterisks (*): individuals typed for the mutation, m: mutation carrier and wt: wildtype. To protect the confidentiality of these results, the genotypes of some unaffected individuals of families A and D are not shown.

#FIG. 2 shows pedigree structure of smaller families with mutations. Blackened symbols denote affected family members; asterisks (*): individuals typed for the mutation, m: mutation carrier and wt: wildtype. To protect the confidentiality of these results, the genotypes of some unaffected individuals of family 469 are not shown.

#FIG. 3 shows the three across species highly conserved amino acid substitutions R1441C, Y1699C and I1122T (SEQ ID NOs:21-39).

#Clones:

#1) 4E1 2) 4A11 3) 3G3 4) 7F1 5) 2H8 6) 2D7 7) 4A8 8) 2B5 9) 4G11 10) 3G6 11) 4G11 12) 3D9 13) 4H11 14) 4F12 15) 4B7.

#DNA Extraction