JPND coFUND ADAGE

Late onset Alzheimer's disease (AD) may have a long natural history and the search for peripheral
biomarkers that could be used for screening in the preclinical phase of the disease is a major challenge.
Ageing is the major risk factor for AD and the most recent conceptualizations of ageing and age-related
diseases, including neurodegeneration and AD, indicate that they share basic mechanisms, e.g. (neuro)-
INFLAMMAGING. Thus, to better understand AD pathogenesis and to identify such early biomarkers
capable of distinguishing AD from physiological ageing without dementia, it is mandatory to fully posit AD
within the framework of ageing process and to study LONGITUDINAL COHORTS OF
PRESYMPTOMATIC INDIVIDUALS. The main goal of ADAGE is to identify preclinical circulating
biomarkers deviating from healthy ageing trajectories towards AD. To this aim the project will adopt the
innovative strategy of comparing extreme phenotypes, i.e. blood samples from AD patients collected
years before the clinical onset of the disease versus blood samples from DEMENTIA-FREE
CENTENARIANS (100+) and their offspring (CO), and HEALTHY NONAGENARIAN SIBLINGS (90+
sibs) characterized by familial longevity, who maintain a good cognitive status despite their very
advanced age. Accordingly, ADAGE will exploit unique, large and very informative EXISTING COHORTS
where biomaterials (blood, plasma, serum and brains) are available and whole genome genetic and
epigenetic studies (as well other omics investigations) have been recently performed: i) old TWINS OF
THE SWEDISH TWIN REGISTRY (STR) FOLLOWED LONGITUDINALLY FOR >45 YEARS, assessed
for lifestyle, cognitive status and exposure to toxicants. Here, incident and prevalent cases of AD
DISCORDANT TWINS have been identified, blood/serum/plasma have been collected at different time
points before AD clinical manifestation and post-mortem brains are available; ii) 100+ and their CO as
well as 90+ sibs as gold standard of healthy ageing, who have never shown any signs of cognitive
disability/decline and are fully characterized phenotypically and with a variety of omics, including genetics
and epigenetics. In these cohorts, ADAGE will perform a metanalysis of existing omics data, an analysis
of circulating proteome (several hundred proteins) and metabolome integrated by studies on circulating
2
miRNA, advanced flow cytometry and an in depth neuropathological analysis of AD versus normal old
brains, focused on markers of inflammation/cell senescence, to be correlated with cognitive status and
omic data. In a consistent subgroup of AD patients and controls where both blood and CSF is available
an in depth analysis (proteomics, metabolomics) of the two fluids will be performed. representing a. This
approach will allow to validate the results obtained in peripheral specimens in the other cohorts but also
to identify new combined CSF and blood AD signature(s). On the whole, ADAGE has the potential to
identify new molecular biomarkers of cognitive aging as well new risk and protective factors for dementia
on bench to bedside translational level (diagnostic and druggable targets).

UCL has recently developed a multiplexed, MRM-based UPLC-MS/MS assay for the simultaneous
quantitation of 27 biomarkers in 100 ul of CSF in under 10 min (figure below). Using this assay in the
analyses of CSF, it was possible to distinguish correctly between patients with Parkinson's disease,
Alzheimer Disease, Lewy Body Dementia (LBD) and age-matched controls (figure 2 and 3). This test
was developed on a triple quadrupole based mass spectral platform which allows the translation of this
test into any large hospital.
Preliminary metabolomic profiling of CSF in the UCL laboratories have identified potential biomarkers
and differences in metabolite expression in patients with Lewy Body Dementia, Parkinson's Disease and
age-matched controls (Figure below). These results indicate that there are changes in the metabolome
and potentially the lipidome (brain is 60% lipid) of patients with neurodegenerative conditions. Cutting
edge 'omic' techniques, a full translational test capability and well defined clinical patient samples
included in WP3, will allow us to identify new biomarkers, potential disease mechanisms and therapeutic
targets for neurodegenerative conditions.
Ageing, Single-Carbon Metabolism, and Total Brain Atrophy in Non-Demented Elders: Ageing (table 1a),
hyperhomocysteinemia (>10.53)(table Xb), and low folate (<18.67)(table xc) increase risk for total brain
atrophy (Bowman et al., 2012; Bowman et al., 2013). Virtually all brain regions atrophy with age and
these structural changes increase risk for cognitive decline and Alzheimer's disease. These data suggest
that single-carbon metabolism has a role in brain aging.

Impact summary
Everybody grows old. But while some people grow old in good health, others experience severe diseases,
like Alzheimer's disease. Recent research demonstrates that Alzheimer's disease shares several
features with the ageing process and suggests that the pathology is the consequence of something that
went wrong during physiological ageing. Researchers involved in ADAGE (Alzheimer's
Disease+AGEing) will use the most advanced biomolecular approaches to compare Alzheimer's disease
patients with people that have successfully aged, like nonagenariangs and centenarians. In this way they
will identify early biomarkers of the disease that will provide new diagnostic, prognostic and therapeutic
targets.
(b) What are the specific aims of the patient/public involvement
The ADAGE consortium will devote effort to establish intense, regular and long lasting interaction with
patient's associations. This will significantly improve the dissemination of project results among
healthcare and patients communities and contribute to promote the theoretical architecture at the basis
of ADAGE.
(c) How will you perform the PPI - (i.e. what approach to PPI will be used)?
CRIC has been included as external collaborator. CRIC provides massive health care support to
thousands of patients and has intense interactions with patients's associations. CRIC involvement led to
the concrete interaction with the Federazione Veneto Solidarietà Alzheimer, an association that supports
the network of all the AD patients associations of Veneto region, that has officially endorsed ADAGE
proposal