1. COST CM1103 Training School Structure-based drug design for diagnosis and treatment of neurological diseases Istanbul, 9-13 Sept 2013 Mirjana Babić, mag.biol.mol. Laboratory for Developmental Neuropathology Croatian Institute for Brain Research Biomarkers of Alzheimer’s disease

2. “ Detection and tracking of biological markers for early therapeutic intervention in sporadic Alzheimer's disease” Project of the Croatian Science Foundation grant no. 09/16 from 1st Jan 2012 – 31st Dec 2014 neurodegenerative disorder loss of memory and cognitive decline in 2050 - approximately 80 million people will suffer from Alzheimer’s disease Alzheimer's disease

3. Ideal marker for diagnosis of Alzheimer's disease is not found yet! Diagnosis of AD based on criteria of: DSM-IV-TR NINCDS-ADRDA ICD 10 Characteristics of good marker: sensitivity and specificity above 85% availability non invasiveness acceptable price possibility for repetitive measures Aim of this project to determine the diagnostic accuracy of potentially highly useful biological markers for discrimination among subjects mild cognitive impairment (MCI), non-demented HC, and patients with other primary causes of dementia

4. Neuropsychological testing Early detection of non-cognitive BPSD (behavioural and psychological symptoms of dementia): o NPI (Neuropsychiatric Inventory) o ADAS-noncog (Alzheimer's disease Assessment Scale for non-cognitive symptoms) o BEHAVE-AD (behaviour rating scale) Laczo et al., 2009. Additional testing of patients with the risk of AD: o Hidden-goal task (human analogue of the Morris water maze task)

5. Imaging biomarkers Earliest change in the brain of AD patients is atrophy of hippocampus and entorhinal cortex. Monitoring of disease progression by: MRI (Magnetic resonance imaging) MRS (Magnetic resonance spectroscopy) SPECT (Single photon emission computorized tomography) Blennow and Zetterberg, 2006.

6. Genetic biomarkers Gene expression profiling using the RNA extracted from cells precipitated in pellets of CSF samples Familial AD caused by mutations in: 1.APP (amyloid precursor protein) 2.PSEN1 (presenilin 1) 3.PSEN2 (presenilin 2) Sporadic AD 1.ε4 allele of the apolipoprotein E gene (APOE)

7. 1.serotonergic system (5HT-2A, 5HT-1B, 5HT-2C) 2.dopaminergic system (COMT, DBH, MAO-B) 3. inflammation pathways (IL-1, IL-6, IL-10, IL-10, TNF) 4.neuronal development and differentiation (BDNF) 5.lipoproteins’ metabolism (ApoE) Specific polymorphisms of genes coding for components of:

8. CSF biomarkers CSF amyloid β 1-42, total tau and phosphorylated tau are the main reflect two major neuropathological hallmarks of AD - neurofibrillary tangles and senile plaques. T-tau 300% increased in AD patients Aβ 1-42 50% decreased AD patients Andreasson et al., 2007.

9. Phospho-tau reflects phosphorylation state of tau protein and formation of neurofibrillary tangles in the brain P-tau199P-tau181P-tau231 Novel CSF biomarkers VILIP-1, neuronal calcium-sensor protein VILIP-1/Aβ 1-42 ratio sphingolipids

10. Standardization of procedures in CSF analysis Levels of CSF biomarkers vary among different laboratories. The cause are variations in: 1.Pre-analytical procedures 2.analytical procedures 3.differences between ELISA kits of various manufacturers

11. Ultimate goal of this project is to predict AD in healthy, asymptomatic subjects

12. Acknowledgements Thank you for your attention! Please visit: http://alzbiotrack.hiim.hr/