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Colombia Médica

On-line version ISSN 1657-9534

Colomb. Med. vol.46 no.3 Cali July/Sept. 2015

 

Original Article

Analysis of the LRRK2 p.G2019S mutation in Colombian Parkinson's Disease Patients

Análisis de la mutación p.G2019S del gen LRRK2 en pacientes colombianos con enfermedad de Parkinson

Andrés Felipe Duque1, Juan Carlos Lopez1,2, Bruno Benitez1, Helena Hernandez1, Juan José Yunis1,2, William Fernandez1,3, Humberto Arboleda1,4, Gonzalo Arboleda1,2.

1 Grupo de Neurociencias. Facultad de Medicina, Universidad Nacional de Colombia, Bogotá, Colombia
2 Departamento de Patología. Facultad de Medicina, Universidad Nacional de Colombia, Bogotá, Colombia
3 Departamento de Medicina Interna. Facultad de Medicina, Universidad Nacional de Colombia, Bogotá, Colombia
4 Departamento de Pediatría. Facultad de Medicina, Universidad Nacional de Colombia, Bogotá, Colombia

Duque AF, Lopez JC, Benitez B, Hernandez H, Yunis JJ, Fernandez W, Arboleda H, Arboleda G. Analysis of the LRRK2 p.G2019S mutation in Colombian Parkinson's Disease Patients. Colomb Med. 2015; 46(3): 117-121.

© 2015. Universidad del Valle. This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Article history: Received: 04 April 2014 Revised: 02 May 2015 Accepted:21 September 2015

Corresponding author:

Gonzalo Arboleda. Departamento de Patología, Universidad Nacional de Colombia, Bogotá, Colombia. Tel: +57 1 3165000 ext 11613; Fax: +57 1 3681517. E-mail: gharboledab@unal.edu.co


Abstract

Introduction: Mutations in the leucine-rich repeat kinase 2 gene (LRRK2 or Dardarin) are considered to be a common cause of autosomal dominant and sporadic Parkinson's disease, but the prevalence of these mutations varies among populations.
Objective: To analyzed the frequency of the LRRK2 p.G2019S mutation (c.6055G>A transition) in a sample of Colombian patients.
Methods: In the present study we have analyzed the frequency of the LRRK2 p.G2019S mutation in 154 patients with familial or sporadic Parkinson Disease, including early and late onset patients, and 162 normal controls.
Results: Our results show occurrence of this mutation in two cases (2/154, 1.3%) with classical Parkinson's signs, and one completely asymptomatic control (1/162, 0.6%).
Conclusion: The p.G2019S mutation is not an important causal factor of Parkinson Disease in Colombia having similar frequencies to those reported in other Latin American populations.

Keywords:

Parkinson's disease, dardarin, LRRK2, p.G2019S mutation, Colombia.


Resumen

Introducción: Las mutaciones en el LRRK2 (del inglés gen leucinerich repeat kinase 2) o Dardarina se consideran una causa común de enfermedad de Parkinson autosómica dominante. Sin embargo, la prevalencia de estas mutaciones varia en diferentes poblaciones.
Objetivo: Snalizar la frecuencia de la mutación p.G2019S (transición c.6055 G>A) del gen LRRK2en una muestra de pacientes colombianos.
Métodos: En el presente estudio analizamos la frecuencia de la mutación en 154 pacientes con enfermedad de Parkinson familiar o esporádica, y 162 controles normales.
Resultados: Se determinó la presencia de la mutación en 2 casos de Parkinson (2/154, 1.3%) los cuales presentan los signos clásicos de la enfermedad y en un control completamente asintomático (1/162, 0.6%).
Conclusión: La mutación p.G2019S no es un factor causal importante de la Enfermedad de Parkinson en la población Colombiana, y muestra frecuencias similares a las reportadas en otras poblaciones latinoamericanas.

Palabras clave:

Enfermedad de Parkinson; dardarin;LRRK2; mutación; G2019S; Colombia.


Introduction

Parkinson's disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease (AD). PD affects all racial groups and has a fairly uniform worldwide distribution1;2. In Europe, the prevalence of PD is 1.6/1003, while in Colombia the prevalence is 4.7/1,0004.

The predominant feature of PD is the progressive slowness of movement (bradikinesia). Patients initially experience resting tremor, difficulty in the initiation of the movements in the absence of paralysis (akinesia), and muscular rigidity, finally leading to postural instability5. A subset of patients also develops dementia6,7. The main pathological features of PD are the selective and progressive loss of dopaminergic neurons in the substantia nigra compact zone of the midbrain, degeneration of the nigrostriatal system and the presence of Lewy bodies8.

The contribution of genetic factors to the development of PD has been largely underestimated for many yrs, although as many as 15% of PD cases have a family history of the disease9. With the recent description of mutations in several genes associated with earlyonset autosomal dominant and recessive familial Parkinsonism, it has been suggested that PD can have a genetic basis10,11. However, the extent of the genetic contribution and the interaction between environmental and genetic factors remain unclear12.

To date, mutations in several genes have been associated to PD: PARK 1 to PARK 15. Some of these mutations are linked to familial autosomal dominant (PARK 1/4: alpha-synuclein, PARK 8: LRRK2) or recessive (PARK 2: Parkin, PARK 6: DJ-1 and PARK 7: PINK1, PARK 13: ATP13A2) forms of PD10,13,14.

Recently, diverse studies in different populations have identified mutations in leucine-rich repeat kinase 2 gene (PARK 8: LRRK2 or Dardarin) in autosomal dominant and sporadic forms of PD15,16. LRRK2 mutations cause PD in many different populations and their prevalence in familial and sporadic cases varies across these groups17,18. Studies of the frequency of LRRK2 mutations in Latin American countries are scant. In Brazil the prevalence of the LRRK2 p.G2019S mutation is about 2%, being more prevalent among familial than in sporadic PD cases (8.7% vs 0.76%)19. In Chilean population the prevalence of this mutation is around 3%20.

In the present paper we have analyzed the prevalence of the LRRK2 p.G2019S mutation in a sample of unrelated Caucasian-mestizo PD patients from Colombia.

Materials and Methods

We analyzed the frequency of the p.G2019S mutation (c.6055 G>A transition) in a case-control study of 154 PD patients and 162 controls. PD cases were 92 males and 62 females (mean age: 59.5 yrs; SD: 15.0); controls were 60 males and 102 females (mean age: 74.9 yrs; SD: 12.3). Twenty eight patients had early onset PD (less than 40 yrs old: mean age at onset: 29.4 yrs; SD: 7.7). There were 126 late onset PD cases (mean age at onset: 58.2 yrs; SD: 10.5). 130 cases were sporadic (mean onset age: 53.4 yrs; SD: 14.8) and 24 were familial cases (mean onset age: 48.7 yrs; SD: 16.6). Patients and controls were analyzed by an interdisciplinary group at the Movement Disorders Clinic of the National University of Colombia from 2005 to 2014. Diagnosis was done following the United Kingdom Parkinson's Disease Society Brain Bank diagnostic criteria (UK PDSBB)21. Controls were free of any motor or cognitive alterations and did not have family history of PD. The research protocol was approved by the ethics committee of the Faculty of Medicine, National University of Colombia and both patients and controls signed informed consent authorizing their participation in this study.

DNA was extracted from peripheral blood with the Wizard Genomics DNA isolation Kit (Promega Corporation, Madison WI). LRRK2 mutation was genotyped following the methodology described by Nichols16. Genomic DNA was amplified using forward (5'-TTTTGATGCTTGACATAGTGGAC-3') and reverse (5'-CACATCTGAGGTCAGTGGTTATC-3') primers, in an amplification reaction containing 50 ng of DNA, 25 pmol of each primer, 250 mM of each dNTP, 1.5 mM of MgCl2, 2.5 mL of reaction buffer 10X and 1U of Taq polymerase, in a final volume of 25 mL. The amplification conditions were the following: an initial denaturing step at 94° C for 5 min followed by 35 cycles of 94° C for 30 s, 60° C for 30 s, 72° C for 45 s, and a final extension of 10 min at 72° C. The PCR products were then subjected to restriction endonuclease digestion analysis using Sfc1 enzyme (New England BioLabs, USA), resolved in a 70% NuSieve and 30% SeaKeam agarose gel electrophoresis and visualized with ethidium bromide under ultraviolet illumination. The amplified fragments were of 228 and 101 bp for non-mutated, and 228, 207, 101 and 21 bp fragments for the mutant heterozygotes. Sequencing was done using BigDye terminator following manufacturer's recommendations and analysed by an ABI-PRISM 3500 automatic sequencer (Applied Biosystems, USA). NovoSNP software was used to reveal the mutations.

Results

The frequency of LRRK2 p.G2019S mutation was 1.3% in patients (2/154) and 0.6% in controls (1/162), p value= 0.9777. The two LRRK2 p.G2019S mutation cases were heterozygotes for the mutation and exhibited signs of classical PD (Table 1). They both had good response to L-Dopa and experienced L-Dopa induced dyskinesias. The control with the LRRK2 p.G2019S mutation has neither motor alteration nor cognitive impairment at the age of 77 yrs, and had no family history of PD.

Table 1. Clinical features of the LRRK2 G2019S positive cases.
Features Patient 1 Patient 2 Control
Sex Female Male Male
Ethnic origin CM CM CM
Family history no yes* no
Age at exam (yrs) 79 67 77
Age at onset (yrs) 71 60 n/a
Bradykinesia yes yes no
Resting Tremor yes no no
Rigidity yes yes no
Postural Instability yes yes no
Asymmetry at onset yes yes no
Dystonia no no no
Behavioral alterations no no no
L-DOPA response yes yes n/a
Cognitive decline no no no
L-DOPA induced dyskinesia yes yes n/a
CM= caucasian mestizo
*affected both mother and half-brother
n/a= non applicable

Family study of one of the positive patients for the LRRK2 p.G2019S mutation (without family history for the disease) revealed the presence of the mutation in 3 out of 4 of her descendants, none of them have been affected by the disease taking into account that it has been described an age-dependent penetrance of the LRRK2 p.G2019S and the oldest of them was 42 yrs old at the moment of the examination, followed by a male aged 40, a woman aged 39, and the youngest aged 36. All of them are heterozygotes (Figs. 1A and 2). The family of the other LRRK2 p.G2019S positive patient, who had family history of PD, refused to participate in the study (Fig. 1B). The control positive for the mutation had no descendants, neither brothers.

Although the LRRK2 pG2019S mutation is an infrequent causal factor for PD among Colombian population, there is a tendency for increased risk (OR= 2.118, CI95%= 0.190-23.616), that is greatly underpowered to consider it significant.

Discussion

The etiology of PD is complex and has not been completely clarified10,13 Mutations in LRRK2 have been described in different populations, with variation in its frequency. The most common of the LRRK2 mutations is the p.G2019S. The frequency of this mutation is high among North Africans (41%)18, followed by Ashkenazi Jews (18.3%)22, in Europe and North America is around 0.8-6%23-25, while is lower among Latin Americans19,20,26 and absent in Asians27 (Table 2). The most plausible explanation for the observed differences in frequencies is the inherent genetic background of each sample, being Latin-America and Colombia in particular a highly mix population28, that defines their specific susceptibility to the development of the disease. In fact, the mutation has been traced among different populations and the findings raise the possibility of a common ancestor for this mutation, following what might be considered a demographic and geographic pattern of evolution29.

Table 2. Frequency of the LRRK2 G2019S in diverse populations around the world.
Population p.G2019S Frequency (%) Reference
Chile 5/166 (3.0) Perez-Pastene et al.20
Brazil 3/154 (2.0) Pimentel et al.19
Brazil 3/83 (3.5) Munhoz et al.26
Spain 16/302 (5.3) Gaig et al.31
Spain 8/105 (7.6) Infante et al.32
Spain 5/225 (2.7) Mata et al.33
Italy 5/513 (0.8) De Rosa et al.25
Uruguay and Peru Uruguay 5/125 (0.04); Peru 1/240 (0.4) Mata et al.29
Argentina 3/55 (5.5) Gatto et al.30
North African Arabs 4/17 (41.0) Lesage et al.18
Azkenashi Jews 22/120 (18.3) Ozelius et al.22
Asian populations 0/675 (0.0) Tan et al.27
Colombia 2/154 (1.3) Present study

The frequency of the LRRK2 p.G2019S mutation in the Colombian population is one of the lowest reported in Latin American populations and is one of the lowest in the world. Reports from Peru, Chile and Brazil have described frequencies of this mutation between 0.4-3.5%19,20,30, and higher frequency in Argentina and Uruguay (4-5%)30,31. However, the small differences observed in the frequency of this mutation between the present study and other Latin American and European populations studied are more probably related to methodological issues such as sample size and composition (number of familiar and sporadic cases in each study).

Clinical characteristics of the positive LRRK2 p.G2019S patients do not differ from any other PD cases, coinciding with the phenotype described for LRRK2 p.G2019S carriers in other reports15,32,33,34. The control positive for the mutation was completely asymptomatic at the age of 77 yrs. No motor alteration, no cognitive decline and no signs of any neurological disease were found. This could be explained by the incomplete penetrance of the mutation as previously described for carriers of the LRRK2 pG2019S over the age of 70 yrs15,32.

Conclusion

Our results demonstrate that the LRRK2 p.G2019S mutation is an infrequent cause of PD in the present sample, having similar frequencies to other Latin American populations.

Conflict of Interest:

Authors declare that do not have any conflict of interest.

Acknowledgment:

This work was supported by grants from DIB (822002) Universidad Nacional de Colombia, COLCIENCIAS (1101-04-10161) and Banco de la República (No 1,762).


References

1. Khandhar SM, Marks WJ. Epidemiology of Parkinson's disease. Dis Mon. 2007; 53(4): 200-5.         [ Links ]

2. Zhang ZX, Roman GC. Worldwide occurrence of Parkinson's disease: an updated review. Neuroepidemiology. 1993; 12(4): 195-208.         [ Links ]

3. de Rijk MC, Tzourio C, Breteler MM, Dartigues JF, Amaducci L, Lopez-Pousa S, et al. Prevalence of parkinsonism and Parkinson's disease in Europe: the EUROPARKINSON Collaborative Study. European community concerted action on the epidemiology of Parkinson's disease. J Neurol Neurosurg Psychiatry. 1997; 62(1): 10-5.         [ Links ]

4. Pradilla AG, Vesga ABE, León-Sarmiento FE, grupo GENECO. Estudio neuroepidemiológico nacional EPINEURO) colombiano. Pan Am J Public Health. 2003; 14(2): 104-11.         [ Links ]

5. Jankovic J. Parkinson's disease: clinical features and diagnosis. J Neurol Neurosurg Psychiatry. 2008; 79(4): 368-76.         [ Links ]

6. Hughes AJ, Daniel SE, Kilford L, Lees AJ. Accuracy of clinical diagnosis of idiopathic Parkinson's disease: a clinico-pathological study of 100 cases. J Neurol Neurosurg Psychiatry. 1992; 55(3): 181-4.         [ Links ]

7. Vermersch P, Delacourte A, Javoy-Agid F, Hauw JJ, Agid Y. Dementia in Parkinson's disease: biochemical evidence for cortical involvement using the immunodetection of abnormal Tau proteins. Ann Neurol. 1993; 33(5): 445-50.         [ Links ]

8. Jellinger KA. Recent developments in the pathology of Parkinson's disease. J Neural Transm Suppl. 2002;(62): 347-76.         [ Links ]

9. Elbaz A, Grigoletto F, Baldereschi M, Breteler MM, Manubens-Bertran JM, Lopez-Pousa S, et al. Familial aggregation of Parkinson's disease: a population-based case-control study in Europe. EUROPARKINSON Study Group. Neurology. 1999; 52(9): 1876-82.         [ Links ]

10. Dawson TM, Dawson VL. Molecular pathways of neurodegeneration in Parkinson's disease. Science. 2003; 302(5646): 819-22.         [ Links ]

11. Zhang Y, Dawson VL, Dawson TM. Oxidative stress and genetics in the pathogenesis of Parkinson's disease. Neurobiol Dis. 2000; 7(4): 240-50.         [ Links ]

12. Riess O, Kruger R. Parkinson's disease--a multifactorial neurodegenerative disorder. J Neural Transm Suppl. 1999; 56: 113-25.         [ Links ]

13. Douglas MR, Lewthwaite AJ, Nicholl DJ. Genetics of Parkinson's disease and parkinsonism. Expert Rev Neurother. 2007; 7(6): 657-66.         [ Links ]

14. Piccini P, Burn DJ, Ceravolo R, Maraganore D, Brooks DJ. The role of inheritance in sporadic Parkinson's disease: evidence from a longitudinal study of dopaminergic function in twins. Ann Neurol. 1999; 45(5): 577-82.         [ Links ]

15. Healy DG, Falchi M, O'Sullivan SS, Bonifati V, Durr A, Bressman S, et al. Phenotype, genotype, and worldwide genetic penetrance of LRRK2-associated Parkinson's disease: a case-control study. Lancet Neurol. 2008; 7(7): 583-90.         [ Links ]

16. Nichols WC, Pankratz N, Hernandez D, Paisan-Ruiz C, Jain S, Halter CA, et al. Genetic screening for a single common LRRK2 mutation in familial Parkinson's disease. Lancet. 2005; 365(9457): 410-2.         [ Links ]

17. Ishihara L, Gibson RA, Warren L, Amouri R, Lyons K, Wielinski C, et al. Screening for Lrrk2 G2019S and clinical comparison of Tunisian and North American Caucasian Parkinson's disease families. Mov Disord. 2007; 22(1): 55-61.         [ Links ]

18. Lesage S, Ibanez P, Lohmann E, Pollak P, Tison F, Tazir M, et al. G2019S LRRK2 mutation in French and North African families with Parkinson's disease. Ann Neurol. 2005; 58(5): 784-7.         [ Links ]

19. Pimentel MM, Moura KC, Abdalla CB, Pereira JS, de Rosso AL, Nicaretta DH, et al. A study of LRRK2 mutations and Parkinson's disease in Brazil. Neurosci Lett. 2008; 433(1): 17-21.         [ Links ]

20. Perez-Pastene C, Cobb SA, Diaz-Grez F, Hulihan MM, Miranda M, Venegas P, et al. Lrrk2 mutations in South America: A study of Chilean Parkinson's disease. Neurosci Lett. 2007; 422(3): 193-7.         [ Links ]

21. Daniel SE, Lees AJ. Parkinson's Disease Society Brain Bank, London: overview and research. J Neural Transm Suppl. 1993; 39: 165-72.         [ Links ]

22. Ozelius LJ, Senthil G, Saunders-Pullman R, Ohmann E, Deligtisch A, Tagliati M, et al. LRRK2 G2019S as a cause of Parkinson's disease in Ashkenazi Jews. N Engl J Med. 2006; 354(4): 424-5.         [ Links ]

23. Kachergus J, Mata IF, Hulihan M, Taylor JP, Lincoln S, Aasly J, et al. Identification of a novel LRRK2 mutation linked to autosomal dominant parkinsonism: evidence of a common founder across European populations. Am J Hum Genet. 2005; 76(4): 672-80.         [ Links ]

24. Kay DM, Zabetian CP, Factor SA, Nutt JG, Samii A, Griffith A, et al. Parkinson's disease and LRRK2: frequency of a common mutation in U.S. movement disorder clinics. Mov Disord. 2006; 21(4): 519-23.         [ Links ]        [ Links ]

26. Munhoz RP, Wakutani Y, Marras C, Teive HA, Raskin S, Werneck LC, et al. The G2019S LRRK2 mutation in Brazilian patients with Parkinson's disease: phenotype in monozygotic twins. Mov Disord. 2008; 23(2): 290-4.         [ Links ]

27. Tan EK, Shen H, Tan LC, Farrer M, Yew K, Chua E, et al. The G2019S LRRK2 mutation is uncommon in an Asian cohort of Parkinson's disease patients. Neurosci Lett. 2005; 384(3): 327-9.         [ Links ]

28. Ruiz-Linares A, Adhikari K, Acuña-Alonzo V, Quinto-Sanchez M, Jaramillo C, Arias W, et al. Admixture in Latin America: geographic structure, phenotypic diversity and self-perception of ancestry based on 7,342 individuals. PLoS Genet. 2014; 10(9): 1004572.         [ Links ]

29. Zabetian CP, Hutter CM, Yearout D, Lopez AN, Factor SA, Griffith A, et al. LRRK2 G2019S in families with Parkinson disease who originated from Europe and the Middle East: evidence of two distinct founding events beginning two millennia ago. Am J Hum Genet. 2006; 79(4): 752-8.         [ Links ]

30. Mata IF, Cosentino C, Marca V, Torres L, Mazzetti P, Ortega O, et al. LRRK2 mutations in patients with Parkinson's disease from Peru and Uruguay. Parkinsonism Relat Disord. 2009; 15(5): 370-3.         [ Links ]

31. Gatto EM, Parisi V, Converso DP, Poderoso JJ, Carreras MC, Martí-Massó JF, et al. The LRRK2 G2019S mutation in a series of Argentinean patients with Parkinson's disease: clinical and demographic characteristics. Neurosci Lett. 2013; 537: 1-5.         [ Links ]

32. Gaig C, Ezquerra M, Marti MJ, Munoz E, Valldeoriola F, Tolosa E. LRRK2 mutations in Spanish patients with Parkinson disease: frequency, clinical features, and incomplete penetrance. Arch Neurol. 2006; 63(3): 377-82.         [ Links ]

33. Infante J, Rodriguez E, Combarros O, Mateo I, Fontalba A, Pascual J, et al. LRRK2 G2019S is a common mutation in Spanish patients with late-onset Parkinson's disease. Neurosci Lett. 2006; 395(3): 224-6.         [ Links ]

34. Mata IF, Ross OA, Kachergus J, Huerta C, Ribacoba R, Moris G, et al. LRRK2 mutations are a common cause of Parkinson's disease in Spain. Eur J Neurol. 2006; 13(4): 391-4.         [ Links ]