Neuropathological evaluation and apolipoprotein E gene polymorphism analysis in diffuse Lewy body disease

JOURNAL

OF THE

NEUROLOGICAL SCIENCES ELSEVIER

Journal of the Neurological Sciences 136(19%) 140-142

Neuropathological evaluation and apolipoprotein E gene polymorphism analysis in diffuse Lewy body disease Chiaki Kawanishi ap* , Kyoko Suzuki a, Toshinari Odawara a, Eizo Iseki a, Hideki Onishi a, Tomohiro Miyakawa a, Yoshiteru Yamada a, Kenji Kosaka a, Naoki Kondo b, Takayuki Yamamoto b a Department of Psychiatry. Yokohama City University, School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama 236, Japan b Fukushimura Hospital, 19-16 Noyori-cyo, Aza-Yamanaka, Toyohashi 441, Japan

Received 5 September 1995;accepted24 September 1995

Abstract We performed a quantitative neuropathological investigation of senile plaques and neurofibrillary tangles in the brains of 14 Japanese patients with diffuse Lewy body disease (DLBD), and examined apolipoprotein E (APOE) gene polymorphism in these patients. Most DLBD brains had as many senile plaques as those with Alzheimer-type dementia (ATD), but fewer neurofibrillary tangles. The APOE ~4 allele frequency in DLBD was 39.3%, similar to that previously reported in pathologically diagnosed ATD. DLBD and ATD are clinically and pathologically distinct, but may have common mechanisms with regard to the formation of amyloid deposition based on these findings. Future investigation of the pure form of DLBD may clarify the association between Lewy body dementia and APOE. Keywords: Diffuse Lewy body disease;Lewy body; Apolipoprotcin E, APOE r4 allele; Alzheimer-type dementia; Parkinson’s disease;Dementia

1. Introduction Diffuse Lewy body disease (DLBD) is a neurodegenerative disease characterized clinically by progressive dementia frequently accompanied by parkinsonism and psychiatric symptoms. Neuropathologically, DLBD is characterized by the presence of numerous Lewy bodies in the central nervous system (Kosaka et al., 1984). DLBD is divided into two clinicopathological forms (the common form and the pure form), mainly according to the degree of co-existing senile changes (senile plaques and neurofibrillary tangles) in the brain. In the common form, senile plaques (SPs) are found widely distributed throughout the cerebral cortex with mild to moderate neurofibrillary tangles (NFTs) restricted to the hippocampus and the parahippocampal cortex. In the pure form, both senile changes are mild and within the range of physiological aging (Odawara et al., 1994). Recent molecular studies have revealed that the frequency of APOE ~4 is significantly increased in both late onset familial and sporadic ATD patients. A similar associ-

* Correspondingauthor.Tel.: (81) 45-787-2666,Fax: (81) 45-783-2540. 0022-510X/%/$15.00 0 1996 Elsevier Science B.V. All rights reserved SSDI 0022-5 10X(95)0031 2-6

ation of APOE polymorphism has been reported in Lewy body-related disorders including senile dementia of the Lewy body type (Harrington et al., 19941, the Lewy body variant of Alzheimer’s disease (Hansen et al., 1994) and Parkinson’s disease (PD) with dementia. In this study, we evaluated the neuropathological stage of senile changes and APOE polymorphism in 14 pathologically diagnosed Japanese cases of DLBD.

2. Materials

and methods

Senile changes were examined in the brains from 14 cases of DLBD who had more than 10 cortical Lewy bodies in each field of the sites of predilection at a X 100 magnification in preparations stained with an anti-ubiquitin antibody. Thirteen of the cases were diagnosed as the common form of DLBD and 1 case (case l>, as the pure form (Table 1). A quantitative analysis of SPs and ms was performed by a method described previously (Odawara et al., 1994). We used a modification of the staging of SPs and NFTs defined by Odawara et al. (1994).

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of the Neurological Sciences 136 (1996) 140-142

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Table 1 Individual data from 14 Japanese patients with DLBD Case

1 2 3 4 5 6 7 8 9 10 11 12 13 14

Sex

F F F M M F M M M M F M F M

Age at death

Duration of ilhtess(years)

Brain weight(g)

Senile changes SPs stage a

NFT’s stage b

48 75 76 78 78 78 79 80 81 82 82 84 86 88

11 5 2 1 7 8 3 1 I 3 16 4 10 1

1060 1140 1040 1300 1230 1130 1165 1355 1120 1150 1100 1060 1050 1145

* +++ +++ +++ +++ +++ +++ +++ + ++ ++ +++ +++ +++

+ + + + + + ++ + + + f + + ++

Apo E genotype

3/3 3/4 214 3/4 3/4 3/4 3/4 214 3/4 3/4 3/4 3/3 214 3/3

a SPs were counted in the hippocampus, parahippocampus, insula, inferior parietal lobule, precentral motor area and the posterior cingulate cortex. The greatest number (GN) of SPs in any given microscopic field was determined and those in each area were averaged (mean greatest number: MGN). f , O/mm* < MGN < 20/mm2; + , 20 5 MGN < 60 and relatively more SPs in every area, except for parahippocampus; + + + , 60 5 MGN (@lawam et al., 1994). b NRs were counted in the transentorhinal pre-a (TP) region and in the inferior temporal (ITG). f , GN in TP < 9/mm2; + , 9 I GN in TP<60; ++,6O
Genomic DNA was extracted from paraffin embedded brain tissue by the standard method. A polymerase chain reaction was performed to amplify the APOE gene with a set of primers, and the amplified fragments were digested with a restriction enzyme, HhaI (Wenham et al., 1991). Electrophoresis was performed on 6% NuSieve GTG gels (FMC Bio Products), followed by visualization using ethidium bromide staining.

3. Results

Data from the individual DLBD cases are shown in Table 1. In case 1 with the pure form of DLBD, few SPs were found. Among the 13 cases of the common form of

Table 2 Apo E allele frequencies in DLBD patients in this study compared to those of patients with pathologically confirmed ATD reported to date Alleles(n) DLBD Alzheimer-type dementia

Control

l

2(%)

l 3(%)

4%)

28 9oa

10.7 2.2

50.0 64.4

39.3 33.3

113b 134c 184d 116’ 154d

2 3.0 2 8.6 5

65 64.2 57 76.7 90

33 32.8 41 14.7 5

Controls were nemopathologically examined and selected. a Picketing-Brown et al. ( 1994). b Nelbantoglu et al. ( 1994). ’ Harrington et al. (1994). d Zubenko et al. (1994).

DLBD, 10 exhibited stage + f + SPs, the SPs criteria for ATD. The remaining 3 cases had SPs in an amount that staged them at borderline between physiological aging and ATD. With regard to NFTs staging, case 11 showed that of physiological aging, while the rest were rated as borderline between physiological aging and ATD. These findings confirm previous descriptions of a mild degree of NFTs combined with SPs as numerous as those seen in ATD (Odawara et al., 1994). APOE genotypes are shown in Table 1. The frequencies of the E2/~4 heterozygote and the l 3/~3 homozygote were 21.4% each. The frequency of the l 3/~4 heterozygote was 57.1%. Case 1 with the pure form of DLBD was e3/e3. No l 2/e2 or e4/~4 homozygotes were observed. The allele frequencies of ~2, ~3 and ~4 are shown in Table 2. The ~4 frequency in DLBD was 39.3%. The high ~4 frequency in DLBD in our study is similar to that reported previously for ATD (Nelbantoglu et al., 1994; Pickering-Brown et al., 1994; Zubenko et al., 1994) which used pathologically-diagnosed cases for analysis of allele frequencies.

4. Discussion The borderlines between DLBD, ATD and PD with dementia ‘are clinically and neuropathologically obscure. Some authors look upon DLBD as being within the spectrum of ATD, however, the pure form of DLBD is excluded from the spectrum. We compared DLBD to ATD neuropathologically, and differentiated them on the basis of the apparent disparity between ATD and the common

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form of DLBD in the occurrence of NITS (Odawara et al., 1994). We also differentiated DLBD from ATD, based on presence of ubiquitin positive specific neurites in the hippocampus in brains from DLBD (Iseki et al., submitted). Our neuropathological findings with regard to senile changes and the APOE ~4 allele frequency in DLBD suggest that DLBD (common form) and ATD may share some common pathogenic mechanism for amyloid deposition which is affected by APOE phenotype. The components of Lewy bodies have not been fully investigated; however, Arai et al. (1992) have reported that Lewy bodies contain @unyloid precursor protein. Strittmatter et al. (1993) have demonstrated differences between APOE and E4 in binding ability with synthetic /3-amyloid peptide. They have suggested that those differences are one of many factors which contribute to the heterogeneity of the disease (ATD in their article). Further investigation of the pure form of DLBD might provide insight into the correlation between APOE and the neuropathological features of DLBD and AID. We suggest that it is preferable to use pathologically-confirmed cases of the diseases when assessing correlations between gene polymorphisms and clinical phenotypes. Concerning PD, a high APOE ~4 frequency was reported in PD with dementia. In contrast, some authors have demonstrated that ~4 frequency does not differ between PD patients with or without dementia and controls. These studies employed clinically diagnosed cases rather than pathologically confirmed cases. When using clinically diagnosed cases it is likely that cases of DLBD may have been included in the group of PD patients with dementia, since the pure form of DLBD frequently presents with parkinsonism, followed by dementia. Detailed neuropathological and clinical assessment clarify the effect of APOE on the pathogenesis of these dementing disorders.

Sciences I36 (1996) 140-142

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