Argyrophilic grain disease

Prof. Isidro Ferrer, Institut Neuropatologia, Servei Anatomia Patològica, IDIBELL-Hospital Universitari de Bellvitge, Universitat de Barcelona, CIBERNED, Hospitalet de LLobregat; Spain

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D E F

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AGD: Gallyas

Argyrophilic grains (AGs)

Neurons with pre-tangles

Astrocytes with argyrophilia

Coiled bodies in oligodendrocytes

Ballooned neurons in amygdala

AGD at first: a neuropathological finding in some patients with dementia

AGs and pre-tangles are recognized with severalphospho-specific anti-tau antibodies

ARGYROPHILIC GRAIN DISEASE: DEMENTIA WITH GRAINS

Delineation of a process versus definition of a disease

Braak H. Braak E. Argyrophilic grains Characteristic pathology of cerebral cortex in cases of adult onset dementia withoutAlzheimer changes. Neurosci Lett 1987;76124-127Braak H. Braak E. Cortical and subcortical argyrophilic grains characterize A disease associated withadult onset dementia. Neuropathol Appl Neurobiol 1989;15:13-26Ikeda K, Akiyama H. Kondo H. Haga C. A study of dementia with argyrophilic grains. Possible cytoskeletal abnormality in dendrospinal portion of neurons and oligodendrocytes. Acta Neuropathol 1995;89:409-414Tolnay M, Spillantini MG, Goedert M, Ulrich J, Bangui D, Probst A. Argyrophilic Grain disease: Widespreadhyperphosphorylation of tau protein in limbic neurons. Acta Neuropathol 1997;93:477-484Tolnay M, Probst A. Ballooned neurons expressing alpha B-crystallin as a constant feature of the amygdala in argyrophilic graindisease. Neurosci Lett 1998;246:165-168

AGD: A novel tauopathy

Togo T, Sahara N, Yen SH, Cookson N, Ishizara T, Hutton M, de Silva R, Lees A, Dickson DW. Argyrophilic grain disease is a sporadic 4-repeat tauopaty. J Neuropathol Exp Neurol 2002;61:547-556Tolnay M, Sergeant N, Ghestem A, Chalbot S, de Vos RAI, Jansen Steur ENH, Probst A, Delacourte A. Argyrophilic graindisease and Alzheimer´s disease are distinguished by their different distribution of tau protein isoforms. Acta Neuropathol2002;104:425-434Ferrer I, Barrachina M, Tolnay M, Rey MJ, Vidal N, Carmona M, Blanco R, Puig B. Phosphorylated protein kinases associatedwith neuronal and glial tau deposits in argyrophilic grain disease. Brain Pathol 2003;13:62-78.

Molecular pathology of AGD

Ferrer I, Santpere G, van Leeuwen FW. Argyrophilic grain disease. Brain 2008; 131: 1416-1432

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Grains: Golgi method

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DAGD. A: CA1, AT8; B: laterotuberal, AT8; C: CA1, 4R; D: entorhinal cortex, 4R

AGD. A: CA1; B: dentate gyrus; C: entorhinal cortex; D: amygdala; E: accumbens; F: septum; G: putamen; H: substantia nigra; I: white matter; J: CA1 tau4R; K: EC tau4R; I: amyg, αB-crystallin

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AGD cortical. A: CA1; B: amygdala; C: entorhinal cortex; D: frontal cortex

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Argyrophilic grain (AG) staging

• Stage I: anterior entorhinal cortex; mild involvement of the cortical and basolateral nuclei of the amygdala; mild involvement of the hypothalamic lateral tuberal nucleus.

• Stage II: entorhinal cortex; anterior CA1; transentorhinal cortex; cortical and basolateral nuclei of the amygdala; presubiculum; hypothalamic lateral tuberal nucleus; dentate gyrus.

• Stage III: entorhinal cortex; CA1; perirhinal cortex; presubiculum; amygdala; dentate gyrus; hypothalamic lateral tuberal nucleus; mild involvement of CA2 and CA3; mild involvement of the subiculum; mild involvement of other nuclei of the hypothalamus (i.e. mammillary bodies); mild involvement of the anterior temporal cortex, insular cortex, anterior cingulated gyrus, orbitofrontalcortex, nucleus accumbens, septal nuclei; rare grains in the midbrain.

• Stage IV: moderate to severe additional involvement of the neocortex and brainstem

Clinical symptoms

•Cognitive decline and dementia

•Behavioural abnormalities, personality changes and emotional and mood imbalance

•Episodic memory loss

•Amnesia, irritability and agitation, followed by delusions, dysphoria and apathy

•Mild amnestic cognitive impairment is not uncommon as an initial manifestation of AGD

•A small number of patients present with progressive trans-cortical sensory aphasia, prominent abnormal behaviour and aggression, and moderate to severe cognitive impairment consistent with frontotemporal dementia

The presence of AGD plus mild-moderate AD-type pathology is, probably, the cause of dementia and not just the presence of AGD.

The age of onset is around 70 years old. The duration of the disease is between 4 and 8 years

ARGYROPHILIC GRAIN DISEASE: DEMENTIA WITH GRAINS

Genetics

The disease appears to be sporadic

Not known genetic factors, but for a single hereditary case:

Kovacs GG, Pittman A, Revesz T, Luk C, Lees A, Kiss E, Tariska P, Laszlo L, Molnár K, Molnar MJ, Tolnay M, de Silva R. MAPT S305I mutation: implications for agryrophilic grain disease. Acta Neuropathol 116: 103-118 (2008)

The frequency of apolipoprotein E ε4 (ApoE ε4) allele in AGD is similar to that of the general population. Yet the frequency of ApoE ε2 is higher than that observed in AD and controls

Association of AGD with tau H1 haplotype is confusing, with both positive and negative results (Togo et al., 2002; Miserez et al., 2003).

ARGYROPHILIC GRAIN DISEASE: DEMENTIA WITH GRAINSAssociation of AGD with Alzheimer disease, progressive supranuclear palsy, Corticobasal degeneration and α-synucleinopathies

Martinez-Lage P, Muñoz DG. Prevalence and disease associations of argyrophilic grains of Braak. J Neuropathol Exp Neurol 1997;56:157-164

Braak H, Braak E. Argyrophylic grain disease: Frequency of occurrence in different age categories andneuropthological diagnostic criteria. J Neural Transm 1998;105:801-809

Ikeda K, Akiyama H, Arai T, Matsushita M, Tsuchiya K, Miyazaki H. Clinical aspects of argyrophilic graindisease. Clin Neuropathol 2000;19:278-284

Tolnay M, Monsch AU, Probst A. Argyrophilic grain disease. A frequent dementing disorder in old patients. Adv Exp Med Biol 2001;487:39-58

Togo T, Cookson N, Dickson DW. Argyrophilic grain disease: Neuropathology, frequency in a dementiabrain bank and lack of relationship with apolipoprotein E. Brain Pathol 2002;12:45-52

AGD+PSP. A: s. nigra; B: nucleus of the vagus nerve; C: reticular formation; D: CA1; E: subiculum; F: entorhinal cortex; G: dentate gyrus; H: dentate gyrus; I: amygdala

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AGD+PSP. A: septum; B: accumbens; C: caudate; D: medial putamen; E: mammillary bodies; F: basal forebrain; G: frontal cortex; I: amygdala (tufted)

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AGD+ astrocytic plaques: A: CA1; B: entorhinal cortex; C: amygdala; D: temporal cortex

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ARGYROPHILIC GRAIN DISEASE: DEMENTIA WITH GRAINS

AGD and neurotransmitters

Practically nothing known, excepting

Yamada T, McGeer PL, McGeer EG. Some immunohistochemical features of argyrophilic grain dementia with normal cortical choline acetyltransferase levels but extensive subcortical pathology and markedly reduced dopamine. J Geriatr Psychiatry Neurol 5: 3-13 (1992)

Perez-Buira S, Barrachina M, Rodriguez A, Albasanz JL, Martin M, Ferrer I. Expression levels of adenosine receptors in hippocampus and frontal cortex in argyrophilic grain disease. Neurosci Lett 423:194-199 (2007)

0,2

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1,0

1,4

1,8

C AGD

A1R

pro

tein

leve

ls(A

U) *

AGDC

Hippocampus

A1R (37 KDa)

β-Actin (45 KDa)

B

A

AGDC

Frontal Cortex

A1R (37 KDa)

β-Actin (45 KDa)

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1,5

2,5

C AGD

A1R

pro

tein

leve

ls(A

U)

B

AGDC

Frontal Cortex

β-Actin (45 KDa)

ACI (120 KDa)

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1,5

2,5

C AGD

AC

I pro

tein

leve

ls(A

U)

A

AGDC

Hippocampus

β-Actin (45 KDa)

ACI (120 KDa)

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1,5

2,5

C AGD

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I pro

tein

leve

ls(A

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Adenosine receptors in the frontal cortex and hippocampus in AGD

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anti-tau phospho-specific antibodies Thr181 (A), Ser214 (B), Ser396 (C), Ser199 (D), Ser231 (E), Ser422 (F), Ser202 (G) and Ser262 (H), and antibody 7.51 (I)

AGD is a tauopathy with tauphosphorylated at different threonine and serine sites

S-IF Total

66

46

S-IF Total

6864

S-IF Total

6864

S-IF Total

6864

tau 7.51 Thr 181 Ser 262 Ser 422

AGD is a 4R tauopathy

4R immunohistochemistry

A B C

Active stress kinases: SAPK/JNK-P and p38-P, and GSK-3β

Several tau-kinases are localized in neurons with pre-tangles and in grains

tau-kinases co-localize with tau-P in neurons with pre-tangles and in grains

Tau 422

TotalS-IF

6864

TotalS-IF

GSK-3β-P

54

66

54

TotalS-IF

CaMKII

TotalS-IF

MAPK/ERK-P

66

4244

38

TotalS-IF

p38-P

TotalS-IF

MEK-1

45

TotalS-IF

ERK2

4250

TotalS-IF

JNK-1

TotalS-IF

GSK-3α/β

66

5147

TotalS-IF

54

66

SAPK-JNK-P

Tau kinases in sarkosyl insoluble fractions and in total homogenates in AGD

NF tau SAPK/JNK-P + tau

SAPK/JNK-P + tau

p38-P + tau

αB-crystallin, phosphorylated neurofilaments, phospho-tau and tau-kinases in ballooned neurons in theamygdala

Tau kinases and phospho-tau are also present in ballooned neurons in the entorhinalcortex and amygdala

++-++++GSK-3β-P

++++ p+++++++++P38-P

++++ p+++++++SAPK/JNK-P

++++ d++-MAPK/ERK-P

------GSK-3β

------JNK-1

------ERK

------MEK

+++++ d+++++++++Ser422

+++ p++++++++Ser396

+±++ d+++++++Ser214

++++ p+++++++Ser201

+++ p+++++++++Thr181

Coiledbodies

AstrocytBalloonedneuron

TanglesPre-tanglesGrain

αB-crystallin is phosphorylated in ballooned neurons

Specific tau truncated forms are usually encountered in AGD and they differ from those seen in AD

746864

62

AGD AGD AD

AA

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thrombin calpain 2 caspase 3

Thrombin is a strong candidate as a causative protease of truncated tau

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A p62

ubiquitin

The ubiquitin-proteasome system (UPS) in AGD

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D E F

Ubiquitin + AT8

Ubiquitin co-localizes with tau in AGs

Blot Ab: anti-Ubi

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220120100

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mar

ker

ctls AGD-pat.

band

N; Human AGD-Ubiquitin

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type of sample

dens

ity ctl

AGD-patol.

Human AGD-Ubiquitin

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band1 band2 band3 band4

type of samplede

nsity ctl

AGD

p<0.0048 p<0.005

* *

Human AGD-Ubiquitin

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band1 band2 band3 band4

type of samplede

nsity ctl

AGD

p<0.0048 p<0.005

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Increased ubiquitination in selected protein bands in AGD hippocampus

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Mutant ubiquitin

B CAnti-CML

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AGD

B123 C

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LR

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GO

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Control

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BlotBlot AbAb:: AntiAnti--CELCEL

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CTL AGD

BlotBlot AbAb:: AntiAnti--CMLCML

220

12080

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BlotBlot AbAb:: AntiAnti--MDALMDAL

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CTL AGD

BlotBlot AbAb:: AntiAnti--DNPDNP

220

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DNP CEL CML MDAL

dens

ity a

djus

ted

to a

ctin

CTL

AGD

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*

CEL CML MDAL0

25

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75

100

125

150CTLAGD

*

GSA AASA0

50

100

150CTLAGD

Oxidative stress and oxidative damage in AGD

Increased expression of oxidative damage as revealed with different markers

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RAGE: receptor ofadvanced glycationend products

Oxidative stress responses in AGD

A, B: controlC, D: AGD

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DC

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SOD1: A, B: control; C, D: AGD. A, C: CA1; B, D: EC SOD2: A: control, B: AGD. CA1

BA

Oxidative stress responses in AGD

Superoxide dismutase 1 and 2: SOD1 and SOD2

CTL AGD

30

Blot Ab: Anti-NDUFS3

70

Blot Ab: Anti-SDHA

48

Blot Ab: Anti-UQCRC2

57

Blot Ab: Anti-MTCO1

57

Blot Ab: Anti-AIF

48

BlotBlot AbAb:: AntiAnti--ββ--ActinActin

BlotBlot AbAb:: AntiAnti--PorinPorin

31

36

Blot Ab: Anti-UCP4

* *

**

A

B0

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1

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NDUFS3 SDHA UQCRC2 MTCO1 UCP4 AIF

dens

ityad

just

edto

porin

CTLAGD

Mitochondrial respiratory complexes and AIF in AGD hippocampus

Decreased expression of proteins of mitochondrial complexes, and increased AIF

tau-P+ mitochondrial porin. No porin localization in grains

36

Blot Ab: Anti-Eif-2α

Blot Ab: Anti-pEif-2α

Blot Ab: Anti-IRE1

105

BlotBlot AbAb:: AntiAnti--ββ--ActinActin

48

Blot Ab: Anti-ATF6

90

Blot Ab: Anti-XBP1

31

36

CTL AGD

Blot Ab: Anti-ATF6 cleaved

50

A

78

Blot Ab: Anti-GRP 78

94

Blot Ab: Anti-GRP 94

Blot Ab: Anti-KDEL

9478

48

Blot Ab: Anti-PDI

BlotBlot AbAb:: AntiAnti--ββ--ActinActin

60

CTL AGDB

Increased expression of endoplasmic reticulum stress markers in AGD hippocampus

tau4R

tau hyperphosphorylation

hyperphosphorylated tau aggregation

tau ubiquination

oxidative stress oxidative responses RAGE SOD1 SOD2

activation of stress kinasesSAPK/JNK and p38

activation of otherkinases: i.e. GSK-3β

p62

ubiquitin

tau/UBB+1

impaired UPS function

mRNA misreading: UBB+1

AGs

pre-tangles

tangles

AGs

tangles

Sequestration of active stress kinases and GSK-3β

trombin in AGs andtangles

tau

truncated tau

aging mitochondrial anomalies

Endoplasmic reticulum stress

Abnormal protein folding