Dr. Adolfo LOPEZ DE MUNAINHospital DONOSTIA22 de Junio de 2006

Implicación de la dardarina en la enfermedad de

Parkinson

Genética de la E de Parkinson 1817-1900

• 1817 J Parkinson: An essay on the shaking palsy. Whitingham & Rowland, London, 1817

• 1881 Charcot JM Paralysie agitante, tremblement sénile, sclérose en plaque. Gaz Hopit Paris 1881; 54: 98-100.1880 PD Leroud. Contribution a l’étude des causes de la paralysie agitante (Thèse de Doctorat, Paris, 1880)

• 1889-1901 W. Gowers A Manual of Diseases of the Nervous System, Blakiston, London 1889-New York 1901. 15% casos tienen historia familiar (¿ambiental o genético?)

Genética de la E. de Parkinson 1900-1950

• 1916-1927 Pandemia de gripe– Poskanzer DC, Schwab RS. Studies inthe epidemiology of

Parkinson’s disease predicting its disappearence as a majore clinical entity by 1980. Trans Am Neurol Ass 1961; 86: 234-245. Predijeron la desaparción de la EP

– RC Duvoisin, M Yahr. Encephalitis and parkisonism. Arch Neurol 1965; 12: 227-239. La mitad de los supervivientes desarrollaron EP entre 5-10 años después de sufrir encefalitis.

• 1926 J Bell, A Clark A pedigree of paralysis agitans. Ann Eugenics 1926; 1: 455-462

• 1937 W Allan. Inheritance of the shaking palsy. Arch Intern Med 1937; 60: 424-436 (EP como entidad AD ¿atrofia olivopontocerebelosa?)

• 1949 H Mjönes. Paralysis agitans. A clinical genetic study. Acta Psychiatr Neurol Scand suppl 54: 1-194. Casos de 9 Hospitales: 40.7% casos “familares”: herencia AD con penetrancia incompleta (60%)

Genética de la E. de Parkinson 1950-1983

• 1962 GG Spellman. Report of familial cases of parkinsonism: evidence of a dominant trait in a patient’s family. J Am Med Assoc 1962; 179: 372-374.

• 1973 Kondo et al: Parkinson’s disease. Clinical analysis and evidence of a multifactorial analysis. Mayo Clin Proc 1973; 465-475. Se reanaliza la casuística de Mjönes y la propia, concluyendo que el origen era multifactorial pero existía un fuerte componente genético

• 1982 A Barbeau, Pourcher E. New data on the genetics of Parkinson’s disease. Can J Neurol Sci 1982; 9: 53-60. Postula que la forma tremórica de inicio precoz tiene un origen genético.

• 1983 JW Langston et al: Chronic parkinsonism in humans due to a product of meperidine-analog synthesis. Science 1983; 219: 979-980. Primer tóxico ambiental capaz de inducir EP.

Table 3. Effect of Broader Definition of Disease in Second Member of a Twin Pair on Pairwise Concordance From: Tanner: JAMA, Volume 281(4).January 27, 1999.341-346

Paisán et cols. Annals of

Neurol. 2005

4 initial families (UGM)

UGM 3: III-19

89 y/o Onset 1995, RH rest tremor

Gait disturbanceOscilations, Dyskinesias.No cognitive impairmentNo depressionGood voice

Levodopa 450 mg.dpramipexole 2.5 mg.d

UGM 5, III-2

• 61 y/o• Onset 2001

– Tremor R-H– Dystonia R foot

• Levodopa 500 mg.d.

UGM 6

15 PD patients from “other families”

72 y-o, Basque (same town)

Onset: 2001, RH rest tremor,

RL clumsiness

Foot dystonia, RH Rt

Levodopa 500 mg.d

Possible related

6 “sporadic” patients

• Patient 5, 80 y/o

• Gait, imbalance, daily falls

• Palilalia, hypophonia

• Treatment:

– Levodopa 750/d

– Cabergoline 4 mg/d

Linkage analysis (III)

LOD score ( =0.0)

Microsateellite

Chromosome Position

(Mb, USCS)) UGM03 UGM04 UGM05 UGM06

PARK1 D4S2460 90.19 -3.06 -0.12 -4.00 -4.86 D4S423 92.83 -3.49 -0.03 -3.83 -5.95 D4S1647 99.79 -2.46 0.04 -4.05 -5.77 D4S2380 96.24 -3.12 0.06 -3.62 -5.11 PARK2 D6S437 158.73 -3.42 -0.17 0.25 -4.16 D6S305 162.16 -3.48 -0.11 -3.96 -5.58 D6S1277 164.26 -3.33 -0.13 -4.11 -5.30 D6S264 166.67 -3.26 -0.13 0.77 -4.87 PARK3 D2S337 61.58 0.86 0.04 -3.63 0.21 D2S441 68.15 -3.26 0.04 -4.24 -2.40 D2S1394 73.05 -3.55 -0.03 -3.73 -2.90 D2S286 75.25 -3.46 -0.04 -3.75 -1.40 PARK4-5 D4S2397 26.93 -2.80 0.23 -2.94 -0.12 D4S230 28.32 -2.77 0.35 -2.93 -0.32 D4S3350 34.00 -3.37 0.24 -0.74 -2.89 D4S405 40.19 -3.42 0.19 -3.81 0.47 PARK6-7 D1S468 3.6 -0.35 -0.13 -0.45 -5.66 D1S244 10.5 -1.51 -0.13 -0.97 -0.71 D1S199 19.7 1.36 -0.17 -4.06 -2.83 D1S2828 21.68 1.38 -0.20 -0.02 -2.87 D1S2734 22.46 1.38 -0.20 -4.04 -3.43

D1S247 30.54 1.58 -3.64 -4.08 -3.24 D1S2845 43.68 -1.98 -0.13 -0.46 -5.61

Loci excluded in the four Basque families

Further refinement of the PARK8 locus in the Basque families

UGM03 Family

UGM05 Family

FAMILY UGM03 Position

(Mb, USCS) Order Recomb Lodscore ---------- ------------------------- 15.52 D12S1303 0.1480 1.08 21.66 D12S1066 0.4820 0.00 29.52 D12S1631 0.0000 2.92 30.86 D1S21698 0.1070 1.32 31.75 D12S1621 0.0000 0.60 32.22 D12S345 0.0000 3.21 37.55 D12S331 0.0000 0.65 39.49 D12S1668 0.5040 0.00 41.09 D12S1653 0.0980 2.04 42.35 D12S1301 0.1090 0.80 45.62 D12S85 0.5110 0.00 44.85 D12S1713 0.3870 0.02 47.48 D12S339 0.1060 1.87 49.32 D12S1635 0.7570 0.59

FAMILY UGM05

Order Recomb Lodscore ------------------------- D12S1303 0.0690 0.38 D12S1066 0.5010 -0.00 D12S1631 0.0000 1.51 D12S1698 0.0000 1.37 D12S1621 0.4990 -0.00 D12S345 0.0000 1.11 D12S331 0.9940 0.27 D12S1668 0.5050 -0.00 D12S1653 0.0000 2.14 D12S1301 0.0000 1.57 D12S85 0.5000 -0.00 D12S1713 1.0000 1.82 D12S339 0.5010 -0.00 D12S1635 0.1230 0.13

FAMILY UGM03 Position

(Mb, USCS) Order Recomb Lodscore ---------- ------------------------- 15.52 D12S1303 0.1480 1.08 21.66 D12S1066 0.4820 0.00 29.52 D12S1631 0.0000 2.92 30.86 D1S21698 0.1070 1.32 31.75 D12S1621 0.0000 0.60 32.22 D12S345 0.0000 3.21 37.55 D12S331 0.0000 0.65 39.49 D12S1668 0.5040 0.00 41.09 D12S1653 0.0980 2.04 42.35 D12S1301 0.1090 0.80 45.62 D12S85 0.5110 0.00 44.85 D12S1713 0.3870 0.02 47.48 D12S339 0.1060 1.87 49.32 D12S1635 0.7570 0.59

FAMILY UGM05

Order Recomb Lodscore ------------------------- D12S1303 0.0690 0.38 D12S1066 0.5010 -0.00 D12S1631 0.0000 1.51 D12S1698 0.0000 1.37 D12S1621 0.4990 -0.00 D12S345 0.0000 1.11 D12S331 0.9940 0.27 D12S1668 0.5050 -0.00 D12S1653 0.0000 2.14 D12S1301 0.0000 1.57 D12S85 0.5000 -0.00 D12S1713 1.0000 1.82 D12S339 0.5010 -0.00 D12S1635 0.1230 0.13

FAMILY UGM03 Position

(Mb, USCS) Order Recomb Lodscore ---------- ------------------------- 15.52 D12S1303 0.1480 1.08 21.66 D12S1066 0.4820 0.00 29.52 D12S1631 0.0000 2.92 30.86 D1S21698 0.1070 1.32 31.75 D12S1621 0.0000 0.60 32.22 D12S345 0.0000 3.21 37.55 D12S331 0.0000 0.65 39.49 D12S1668 0.5040 0.00 41.09 D12S1653 0.0980 2.04 42.35 D12S1301 0.1090 0.80 45.62 D12S85 0.5110 0.00 44.85 D12S1713 0.3870 0.02 47.48 D12S339 0.1060 1.87 49.32 D12S1635 0.7570 0.59

FAMILY UGM05

Order Recomb Lodscore ------------------------- D12S1303 0.0690 0.38 D12S1066 0.5010 -0.00 D12S1631 0.0000 1.51 D12S1698 0.0000 1.37 D12S1621 0.4990 -0.00 D12S345 0.0000 1.11 D12S331 0.9940 0.27 D12S1668 0.5050 -0.00 D12S1653 0.0000 2.14 D12S1301 0.0000 1.57 D12S85 0.5000 -0.00 D12S1713 1.0000 1.82 D12S339 0.5010 -0.00 D12S1635 0.1230 0.13

PARK8 locus: Significant LOD Score (D12S345, 3.21)

Second and third linked area on chromosome 12

BASQUE FAMILY ID IDENTIFIER bp CONSENSUS UGM3 UGM4 UGM5 UGM6

rs10879192 39471471 C C C/T‡ C C

D12S1698 30855986 - 122 126 118 124

D12S1621 31754700 - 191 191 191 191

rs1523118 37515966 - T T C/T‡ T

D12S331 37547321

- 177 177 177 177

rs11169992 37603474

- C C C

rs10876410 37708557

- T A/T‡ T A

rs10876646 37887093 - T T T T

rs10747736 37912177 -

T C T T

rs10876876+ 38011263 -

A - A A

rs11171789+ 38024258 -

T C T T

K543R+ 38031891 -

G A G G

rs10876886+ 38035530 -

C A/C‡ C C

rs11172282 38161804 -

G C/G‡ G G

rs11172541 38229025 -

A - A A

rs10877201 38298564 -

C T/C‡ C C

rs4548690 38475137 -

T C T T

rs7294916 38494630 T T T T T

rs4423249 38554997 T T T T T

rs515205 38689229 A A A/G‡ A A

rs937110 38815159 C C C/G‡ C C

rs7135815 38815163 T T T T T

rs4768224 38947670 T T T T T

rs10784661* 38943803 C C/G‡ C C C

rs10784662* 38944038 G G/A‡ G G G

R1441G* 38990503 C C C C C

M1646T* 39011294 G G G G G

rs12423567 39063583 G G G G G

rs10784616 39117987 C C C C C

rs11612876 39256712 T T - T T/C‡

rs10784800 39386364 C C C C C

rs10879192 39471471 C C C/T‡ C C

D12S1668 39489795 235 235 235 235 235

D12S1653 41093561 - 215 215 203 215

2n

d linked a

rea, 3

.2 M

b

3rd lin

ked a

rea, 2

.6 M

b

Markers used to fine map the candidate interval and determine inter-family shared haplotype and the boundaries of this haplotype.

38306287 38588369 FLJ40126

38438831 38785928 SLC2A13

38905080 39049354 DKFZp434H2111

39073517 39224969 LOC441636

39222629 39250821 MUC19

39588426 39750361 CNTN1

40117841 40254659 PDZRN4

40761917 40826522 LOC283464

40840421 40918264 YAF2

40966980 40968629 LOC400024

40992156 41006174 MADP-1

41006214 41128690 PPHLN1

The genes and the predicted transcripts within the critical interval defined by Basque inter-family recombinants at rs4548690 and D12S1653.

R1441G mutation

• M1646T present in control population:- 0.01 allele frequency in European control population- 0.16 allele frequency in Basque control population

• Analysis of the R1441G mutation failed to detect any carriers• The genetic cause of PD in our autosomal dominant PD families is the R1441G mutation

• R1441G mutation lies in the Ras domain • Ras domain function:

- membrane traffic pathway- membrane targeting

Other LRRK2 families

+

Fam PD4:• East-Indian• Right hand resting tremor• I1371V LRRK2 mutation• Not present in 374 chromosomes

+

Fam 436:• European• T2356I mutation• Not present in 432 chromosomes

delGTAA

+

Fam 466:• European• Unilateral tremor and dystonia• IVS19 + 4 del GTAA variant• Not present in 400 chromosomes

3’

LRRK2 variants present in the 7 families

VARIANTS NUCLEOTIDE CHANGE AMINO ACID CHANGE CONTROLS ALLELE FREQUENCY

rs2131088 IVS4 +38 A>T T(0.08), A (0.92)

rs5797663 IVS4 +80 ->T

rs10878245 C457T (x5) L153L T(0.38), C(0.62)

H275H T825C (x7) H275H C(0.02), T(0.98)

rs732374 IVS8 -160 C>T T(0.35), C(0.65)

rs7955902 IVS10 -10 C>A A(0.35), C(0.65)

rs10784661 IVS14-54 A>G G(0.4), A(0.6)

rs7308720 C1653A (x14) N551K 250 G(0.07), C(0.93)

rs10784462 IVS14 +68 C>G G(0.4), C(0.6)

IVS19 - 22 C>T T(0.1), C(0.9)

IVS 19+4delGTAA IVS 19 + 4delGTAA 0/200

IVS20 -17-/ A A(0.45), _(0.55)

rs10878307 A2167G I723V G(0.05), A(0.95)

rs7966550 T2857C (x22) L953L C(0.11), T(0.89)

IVS27 + 96 C>A A(0.02), C(0.98)

I1371V A4111G (x29) I1371V 0/187

rs7133914 G4193A (x30) R1398H A(0.2), G(0.8)

rs11175964 G4269A (x30) K1423K C(0.2), T(0.8)

IVS30 + 20 -/A _(0.1), A(0.9)

ss48398560 C4624T (x32) P1542S 1/95 T(0.02), C(0.98)

rs1427263 C4872A(x34) G1624G C(0.37), A(0.63)

rs11176013 A4911G (x34) K1637K A(0.45), G(0.55)

ss48398562 T4937C (x34) M1646T C(0.02), T(0.98)

rs11564148 T4939A (x34) S1647T A(0,27), T(0.73)

S1721G A5163G (x35) S1721G G(0.01), A(0.99)

rs7137665 IVS36 +32 C>T C(0.4), T(0.6)

rs10878371 T5457C (x37) G1819G T(0.49), C(0.51)

IVS38 + 35 G>A A(0.1), G(0.9)

ss48398564 G6055A (x41) G2019S /700-2680

ss48398565 A6241G (x42) N2081D 1/93 G(0.02), A(0.98)

rs10878405 G6324A (x43) E2108E A(0.3), G(0.7)

rs11176143 IVS43 + 53 G>A A(0.1), G(0.9)

T2356I C7068T (x48) T2356I 0/450

ss48398567 A7155G (x48) G2385G C(0.13), T(0.87)

rs3761863 T7190C (x49) M2397T T(0.4), C(0.6)

rs4768238 IVS51 + 88 A>G A(0.18), G(0.82)

DKFZp434H2111/LRRK2

- Representing the 73.33% of the LRRK2 mutation carriers here reported.

- It has been established as the most common mutation in North American, North African and European populations.

LRRK2 mutations

12527

T2356I7068nt, x48

IVS19+4delGTAA

I1371V4111nt, x29

I1122V3366nt, x25 R1441G

R1441C4321nt, x31

R1441H4322nt, x31

IVS31 + 3A>G

I2012T6035nt, x41

I2020T6059nt, x41

G2385R7153nt, x48

M1869T5606nt, x38

Y1699C5096nt, x35

G2019S 6055nt, x41

R1941H5822nt, x40

S1096C3287nt, x24

S1228T3683nt, x27

987-1241 1870-20841295-1480 2152-2465

R1067Q3200nt, x24

IVS33 + 6 T>A

WD40

Heat repeat LRR RAS KINASEARM

R793M2378nt, x19

Q930R2789nt, x21

A3342G

R1514Q4541nt, x32

• 767 enfermos afectados de 358 familias– 35 (5%) mutación Gly2019Ser

• 34 heterocigotos, 1 homocigoto

• Discordante en 5 de 19 hermanos (fenocopias)

• 13 (37%) un padre afecto (Baja penetrancia?)

– Clínica típica. Evolución benigna

A frequent LRRK2 ....Di Fonzo et al. Lancet 2005

• 61 familias, 4 (6,6%) con la mutación Gly2019Ser– Italia, Portugal y Brasil

• Clínica habitual: 38 a 68 años, algo más joven

• 4 mutaciones de 41 a 58 aun no enfermos

• 482 enfermos, 263 patología comprobada• 8 (1,6%) mutación Gly2019Ser

– 2 tienen un familiar de primer grado afecto– 41 a 70 años. Clínica típica, 1 distonia

• 3 autopsias– Todos degeneración nigral con C L– Dos con CL corticales

Esporádicos

Padres fallecen antes de contraerla

(58 a 92, m: 74)

Penetrancia baja de la mutación genética

Dificultad del consejo genético

Objective:

255 patients with PD living in Guipúzcoa (Basque country, Spain)

- 168 “sporadic”- 87 “familial”

34 possible autosomal dominant 23 of 4 UGM families

11 of 10 other families 53 AR or no pattern

LRR RAS ARM DOMAIN KINASE WD40HEAT REPEAT DOMAIN

I1122V3366nt,

x25

T2356I7068nt, x48

Y1699C5097nt, x35

R1441GR1441C

4323nt, x31

G2019S6057nt, x41

I2020T6060nt, x41

I1371V4111nt, x29

LRRK2/DARDARIN GENE

“Basque” mutationArg1441Gly

Clinical characteristics of PD- Basque LRRK2/dardarin mutation (PARK8)

Patients:

RESULTS: R1441G present in

• 41/255 (16%) PD pts studied

– 20/23 of UGM families (3 phenocopies)

– 15/64 (21%) of other familiar pts• 2 non Basques families

– 6/168 (3.5%) of “sporadic” patients

Problema con los portadores

Pueden conocer o no el resultado

• Ventajas• Planificacion de su futuro• Terapia neuroprotectora

• Inconvenientes• Angustia

Terapia neuroprotectora

• Rasagilina

• Coenzima Q10

• Ejercicio

Gandhi S et al, 2005

A modo de conclusión (provisional)La Enfermedad de Parkinson: nature or nurture ?• La enfermedad de Parkinson es de origen genético en una

proporción significativa pero aún indeterminada de casos• El fenotipo clínico de la EP genética es indistinguible la EP

esporádica (o considerada como tal)• La variabilidad fenotípica pueden existir incluso dentro de

una misma familia• El patrón de herencia predominantemente hallado hasta

ahora es monogénico y mendeliano, pero la penetrancia variable y la edad tardía de aparición puede enmascararlo

• La enfermedad de Parkinson ofrece unas perspectivas prometedoras para construir una teoría integrada que contemple los factores genéticos primarios, los secundarios y los agentes ambientales

Agradecimientos:

• Coro Paisán (NIH, Bethesda)• Andrew Singleton (NIH, Bethesda)• Jordi Pérez Tur (IBV, Valencia)• José Félix Martí Massó• Todos los miembros del grupo de Neurogenética del Hospital

DONOSTIA• A los pacientes que han colaborado y a sus familiares• Al Hospital Donostia, Nuestra Sra. Antigua y Mendaro• A todos las personas, centros e instituciones que han

colaborado en el estudio