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P2‐231: EXPLORING THE NEED FOR ROBUST BIOMARKER ASSAYS IN ALZHEIMER'S DISEASE AND OTHER NEURODEGENERATIVE DISEASES
Author(s) -
Blennow Kaj,
Dage Jeffrey,
Johnson Sterling C.,
Reiman Eric M.,
Shaw Leslie M.,
Simon Maryline,
Eichenlaub Udo,
Bittner Tobias,
Stutz Vivienne,
Wahl Simone,
Wild Norbert,
Batrla-Utermann Richard,
Molinuevo Jose Luis
Publication year - 2019
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.2019.06.2638
Subject(s) - biomarker , disease , clinical trial , medicine , biomarker discovery , context (archaeology) , neurogranin , bioinformatics , proteomics , biology , paleontology , biochemistry , protein kinase c , gene , enzyme
non-carrier subjects. Total lipid content and oxidation were measured using BODIPY dyes on a subset of lymphocyte samples. AD was diagnosed by McKhann et al. 40 ml of blood was collected in tubes containing acid-citrate-dextrose anticoagulant. Platelet-rich plasma and buffy coat were isolated by centrifugation. Lymphocytes were stained with Mitotracker Red/Annexin V, MitoSox Red, JC-1, or BODIPY and fluorescence was measured via flow cytometry. Mitochondria were isolated from platelets via nitrogen cavitation. Platelet mitochondria were used to spectrophotometrically determine COX and CS Vmax. Values were normalized to protein content. Lymphocyte cultures were expanded up to seven days and harvested for protein, RNA, and DNA. Cultured lymphocytes were also used to determine oxygen consumption with an Oroboros O2K and ATP/ADP levels with a bioluminescent assay. Group means were compared using Student’s T-test or One-Way ANOVA (posthoc LSD). Results: Mean COX Vmax (normalized to protein content) was lower in the APOE ε4 carriers (p<0.05). Lymphocyte Annexin V was significantly higher in APOE ε4 carriers (p<0.05). No differences were observed for MitoSox, JC1, MitoTracker , or BODIPY fluorescence. ATP levels (normalized to protein) were higher in APOE ε4 carrier cultured lymphocytes (p<0.05); no difference was observed in measured oxygen consumption parameters. Protein expression analysis from cultured lymphocytes revealed reduced phosphorylated mTOR, increased phosphorylated SIRT1 and ACC, increased PINK1, AceCS1, and ATP citrate lyase expression. Conclusions: Our data support a relationship between APOE genotype and bioenergetic pathways. Further studies are warranted to understand the relationship between APOE and energy sensing molecular machinery.