invecchiamento e fragilità: il contributo della metabolomicaaging, frailty and age-related diseases...
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Invecchiamento e fragilità:il contributo della metabolomica
Roberta Pastorelli
Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milano
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Metabolomics Definitions
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Metabolites are bio-active small molecules involved in the biochemical
network with a molecule mass less than 1 KDa in size Primary metabolite (endogenous) is directly involved in the normal growth,
development, and reproduction ( e.g. Sugars, Nucleosides, Amino acids,
Lipids)
Secondary metabolite (exogenous) is not directly involved in those processes,
but usually has important ecological function ( e.g. drug, gut microbiota)
Metabolome represents the collection of all metabolites in a given
biological sample
Metabolomics is the systematic identification and quantitation of the
metabolites fingerprint of a biological sample
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‘-OMICS’ sciences
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Environment (lifestyle, drug, diet, age, gender…)
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The Metabolome does not exist in isolation
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano 4
Metabolomics: A Biological Integrator
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
An individual’s metabolic fingerprint
reflects alterations in homeostasis that
underlie health, disease and response.
This fingerprint is also a key tool for
understanding health influences that
operate by changing metabolism,
including:Genetics/Microbiota/Environm
ent/Diet/Epigenetics/Combined effects
Measuring metabolites offers functional understanding of complex biological processes
Given metabolites’ proximity to the phenotype, metabolomics is an important tool
for unlocking the full potential of disease research, genomics and precision
medicine.
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Metabolome Complexity
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Metabolic pathways do not exist in isolation
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Metabolome Complexity
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Metabolites Can Appear in Multiple Pathways
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One technology – many applications
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Not only does metabolomics allow
to extract value from other types of
“omics data”, it can be used to
discover biomarkers, track
treatment effects etc.
And, because the metabolite
composition is central to every
living organism, the in-depth
biological insights metabolomics
provides can advance research
across a variety of areas.:
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Metabolomics strategies
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano 9
Metabolism and Aging
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano 10
Basic concept in metabolism
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano 11
The big picture
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
glycolysis - glucose oxidation in order to
obtain ATP
gluconeogenesis - glucose synthesis
from smaller percursors, to be used by
the brain.
citric acid cycle (Krebs' cycle) - acetyl-
CoA oxidation in order to obtain GTP and
valuable intermediates.
oxidative phosphorylation - disposal of
the electrons released by glycolysis and
citric acid cycle. Much of the energy
released in this process can be stored as
ATP.
urea cycle - disposal of NH4+ in less toxic
forms
fatty acid β-oxidation - fatty acids
breakdown into acetyl-CoA, to be used
by the Krebs' cycle.
gluconeogenesis - glucose synthesis
from smaller percursors, to be used by
the brain.
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The Seven Pillars of Aging
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano 13
Hallmarks of aging
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Mechanistics hallmarks that contribute to ageing to different extents and in
different cells types within an organism. from Lopez-Otin C et al Cell, 2016
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Metabolism and Epigenetic alteration
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Metabolic control of histone methylation
GENE EXPRESSION
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Metabolism & Deregulated nutrient sensing
Invecchiamento e fragilità: il contributo della metabolomicaDott. Roberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Metabolic sensors
mTOR (aminoacids)
AMPK (ADP/AMP) Sirtuins (NAD+/NADH)
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Metabolism & mitochondrial dysfunction
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
TCA cycle decreased efficiency
Inadeguate TCA cycle intermediates
Respiratory chain defects due to
enzyme/substrate alterations
………
from Rustin et al., Mech Age Develop 2000
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Aging, frailty and age-related diseases
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Frailty is a nonspecific state of vulnerability, which
reflects multisystem physiological change
Interrelation between aging and diseases leading to
the development of frailty.
“There is a strong interrelation between
the biological aging changes and the development of
age-related diseases.
The interface could be the development of the-frailty
syndrome.
Besides this intersection, either the biological
aging or the age-related diseases can lead to the
frailty-syndrome characterized by the clinically
apparent changes leading to catastrophic
consequences».
From Fulop et al., 2010.
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The need of multidimensional approach
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano 19
Literature overview of the termsmetabolomics/metabolome and frailty/frail
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
0
1000
2000
3000
4000
5000
6000
2 0 0 0 2 0 0 2 2 0 0 4 2 0 0 6 2 0 0 8 2 0 1 0 2 0 1 2 2 0 1 4 2 0 1 6 2 0 1 8
NU
MB
ER O
F P
UB
BLI
CA
TIO
N
YEAR
(PubMed)
Metabolomics
frailty
Metabolomics & Frailty
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Plasma aminoacids & Frailty
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Circulating aminoacids trajectories linked to malnutrition (both quantitative
and qualitative), a common causative factor of frailty and sarcopenia
From Calvani R et al , 2018
From Sun L et al., 2017
21
Muscle metabolome & Frailty
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
The major differences in muscle
metabolome of frail older subjects to
healthy older subjects relate to
mitochondrial function,
fiber-type composition
tissue turnover
BCCA correlate with genes related to
connective tissue/extracellular matrix
(SPARC)
From Fazelzadeh P et al 2016
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Pre-frailty Metabolomic sub-phenotypes
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Metabolomics allowed discriminating
metabolic sub-phenotypes of pre-frailty
both at the gender level and depending on
the pre-frailty progression and reversibility,
with the identification of specific circulating
discriminant metabolites.
From Pujos-Guillot et al., Frontiers in Physiology, 2019
“In the stable population, pipecolic acid, proline, butyrylcarnitine,
and lysoPEs were found significantly upregulated in pre-frail
men, whereas in women, most of the significant metabolites were
found down regulated. In the group of subjects who will improve
their frailty status (Improvement), the most significant modulated
metabolites are amino acids and derivatives, as well as sugars”
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..still many questions
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Organs influence each other's function via multiple mechanisms including
endocrine signaling, thereby influencing metabolism and ageing in a non-
autonomous fashion.
How do nutrition, exercise and related signaling pathways affect
ageing and frailty?
What metabolites affect ageing (e.g. immune system function) and
frailty/frailty progression?
Are gender differences in frailty affecting metabolome signatures?
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Metabolomics & frailtuning
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
Unit of Proteins and Metabolites Biomarkers,
Lab of Mass Spectrometry
Istituto di Ricerche Farmacologiche Mario Negri IRCCS,
Milano
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Metabolomics & frailtuning
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
WP2. Biomarker assessmentTask 1. Analysis of participants’ metabolic profile
identification of metabolic phenotypes related to
the concept of frailty
The plasma metabolomic profiling are performed by a
targeted mass spectrometry-based quantitative
approach that allows the simultaneous identification
and quantitation of approx. 200 circulating
metabolites spanning over 6 metabolic classes
(acylcarnitines, aminoacids, biogenic amines,
glycerophospholipids, sphingolipids, sugars).
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Metabolomics & frailtuningList of the measurable metabolites
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano 27
Metabolomics & frailtuningPreliminary metabolomic results
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
✓ Frail subjects are separated from pre- and non-
frail subjects mainly by different levels of
aminoacids (increased trend in Fr) and long-
chain diacyl-phosphatidylcholines (PCaa)
(decreased trend in Fr)
✓ Analyses on larger number of subjects and
longitudinal cohort are ON-GOING
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Challange and Hope
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano
define unifying metabolic escape principles that would be suitable to reset metabolism as a means to prevent frailty
29
Invecchiamento e fragilità: il contributo della metabolomicaRoberta Pastorelli/ Istituto Mario Negri IRCCS, MIlano 30