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P4‐228: Techniques for assessing Alzheimer's disease progression
Author(s) -
Budgeon Charley Ann,
Murray Kevin,
Turlach Berwin A.,
Baker David,
Villemagne Victor L.,
Burnham Samantha
Publication year - 2015
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2015.08.057
Subject(s) - disease , longitudinal data , regression , dementia , sigmoid function , polynomial , longitudinal study , regression analysis , statistics , mathematics , computer science , medicine , algorithm , artificial intelligence , data mining , pathology , mathematical analysis , artificial neural network
Background: DYRK1A kinase has been implicated in cognitive impairment and mental retardation of DS individuals who upon aging invariably develop AD-type dementia. We have previously shown that DYRK1A is associated with cytoskeleton through a DYRK1A gene dosage-sensitive mechanism (JNEN 71, 1100, 2012). Both the brain tissue and lymphocytes of DS patients displayed a significant reduction in the yield of actin and tubulin co-immunoprecipitated with DYRK1A antibodies. In the present study, similar approach has been applied to analyze lymphoblastoid cell lines(LCLs) established from AD patients. Methods: Association of cytoskeleton with DYRK1A was examined by co-immunoprecipitation with DYRK1A antibodies and by immunofluorescence staining in six control and twelve AD LCL cell lines purchased from Coriell Cell Repositories, as well as, in DS and fragile X cell lines established at IBR. In addition, Gand F-actin enriched fractions were obtained from these cell lines by ultracentrifugation, TCA precipitation, and then analyzed by Western blotting. Results: Like DS, in the co-immunoprecipitation assay with DYRK1A antibody, all twelve AD LCLs, including sporadic and familial cases, displayed characteristic reduction in the yield of cytoskeletal proteins. On average, the yield of actin was reduced by 50% allowing clear distinction between healthy donors and theirs diseased counterparts. The changes seem to be specific for AD since they were not present in samples prepared from fragile X or unclassified mental retardation cases. Analysis of Gand Factin enriched fractions further revealed another feature distinguishing DS/AD from controls. G-actin fractions of DS and AD consistently displayed a characteristic shift in the electrophoretic mobility of the actin band. The changes are detected before clinical onset of the disease and are the same in DS, familial and sporadic AD cases suggesting a common mechanism of pathogenesis. Conclusions:We have detected similar quantitative and qualitative cytoskeleton alterations in cultured AD LCLs as well in the brain. The presence of alterations in the peripheral tissue links them to the AD disease process and suggests that early dysfunction of cytoskeleton may be at root of Alzheimer-type dementia. The cytoskeleton alterations in LCLs described in our study may potentially be used for AD diagnosis.