Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. Assistant Professor Horticulture & Crop Science Food Science & Technology [email protected]
Tomatoes, Health & the Metabolome
Jessica Cooperstone, Ph.D.
Assistant Professor
Horticulture & Crop Science
Food Science & Technology
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Are tomatoes healthy?
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Better understand the relationship between phytochemicals in plants and health outcomes,
using metabolomics to elucidate these relationships
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Tomatoes and prostate cancer
• Relationship between foods and prostate cancer investigated in the HPFS (Giovannucci et al., J Nat Cancer Inst 1995)
• ↑ lycopene intake (primary source: tomatoes), ↓ total prostate cancer, ↓ ↓ lethal
prostate cancer (Zu et al., J Nat Cancer Inst 2014)
• Meta-analysis finds consistent ↓ risk (RR: 0.81) (Rowles et al., Prostate Cancer and Prostatic
Diseases 2018).
• Pre-clinical data also suggests and chemopreventative effect of tomato
consumption (Boileau et al., J Nat Cancer Inst 2003, Tan et al., Cancer Prev Res 2017)
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
tetra-cis-lycopeneall-trans-lycopene
439 473
503
Nanometers
250 300 350 400 450 500 550 600 650
Ab
so
rba
nce
tetra-cis-lycopene all-trans-
lycopene
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
0
20
40
60
80
100
120
0 2 4 6 8 10 12
Baselin
e c
orr
ecte
d tota
l ly
copene
(nm
ol/L T
RL ±
SE
M)
Hours post meal consumption
Tangerine tomato
Red tomato
On average, lycopene
is 8.5 times more
bioavailable from
tangerine compared to
red tomato juice.
Fractional absorption
tangerine: 47.7 ± 8.81%
red: 4.98 ± 1.92%
cis-lycopene
all-trans-lycopene
Red tomato
Tangerine tomato
Cooperstone et al., Mol Nutr Food Res 2015:59, 658-669
Lycopene is more bioavailable from tangerine tomatoes
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Light micrographs of red and tangerine at 400x magnification.
Carotenoid physical storage form differs between red and tangerine tomatoes
Cooperstone et al., Mol Nutr Food Res 2015:59, 658-669
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Cooperstone et al., Food Chem 2016;210:466-472
Carotenoid physical storage form affects processing stability
tetra-cis-lycopene
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Carotenoids are higher in human plasma after consuming tangerine tomato juice
E
N
R
O
L
L
M
E
N
T
R
A
N
D
O
M
I
Z
E
Control (no juice)
Tangerine tomato juice (2 cans/day, ~325 mL)
Red tomato juice(2 cans/day, ~325 mL)
S
U
R
G
E
R
Y
3-5 weeks
n=15
n=15
n=15
Carotenoids are higher in human plasma after consuming tangerine tomato juice
E
N
R
O
L
L
M
E
N
T
R
A
N
D
O
M
I
Z
E
Control (no juice)
Tangerine tomato juice (2 cans/day, ~325 mL)
Red tomato juice(2 cans/day, ~325 mL)
S
U
R
G
E
R
Y
3-5 weeks
n=15
n=15
n=15
0
500
1000
1500
2000
2500
Visit 1 (baseline) Visit 2 (surgery)
nm
ol/L lyco
pen
e Control
Red
Tangerineab ab bc
b
c
d
Unpublished data, Pelotonia IRPDifferent letters denote statistically significant differences via two-way ANOVA with Tukey’s posthoc test (P<0.05).Different letters denote statistically significant differences via two-way ANOVA with Tukey’s posthoc test (P<0.05).
Unpublished data, Pelotonia IRP
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Can tomatoes modulate prostate cancer in vivo?
AIN-93G (control)
AIN-93G + 10% tangerine tomato powder
AIN-93G + 10% red tomato powder4 week old, male
TRAMP+/- mice
Week 0 Week 18
sacrifice
Carotenoid analysis
Tumor pathology
tetra-cis-
lycopene
Tangerine
all-trans-
lycopene
Red
Unpublished data, AICR grant #318555
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
0
500
1000
1500
2000
2500
3000
3500n
mo
l/L
Tangerine
Red
Plasma carotenoids after 18 weeks of consumption
of 10% tomato powder containing AIN-93G diet
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Control Red Tangerine
High Grade Pin
Microscopic Cancer
Macroscopic Cancer
Most severe cancer pathology observed in each
animal, displayed as a percentage.
Unpublished data, AICR grant #318555
Red and tangerine tomatoes reduce prostate cancer by ~40%
Tangerine tomatoes lead to higher plasma carotenoids
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
0.0
0.5
1.0
1.5
2.0
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4.5
5.0
20 W
ee
ks
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ee
ks
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ks
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ee
ks
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ks
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ks
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ee
ks
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ee
ks
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ks
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ee
ks
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ee
ks
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ee
ks
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ee
ks
33 W
ee
ks
34 W
ee
ks
35 W
ee
ksAve
rag
e n
um
ber
of tu
mo
rs p
er
ma
le m
ou
se
Male Control
Male Tangerine
Male Red
Cooperstone et al., Sci Reports, 2017;7:5106
0
200
400
600
800
1000
1200
1400
1600
Tangerine Red
pm
ol ly
co
pe
ne
/g s
kin
*
Tomatoes can modulate risk for skin cancer as well
*
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
So what is going on here?
• Tomato consumption decreases development of disease (prostate and skin cancer)
• Tumor development seems uncorrelated to carotenoid concentrations in blood or tissue
What else may be exerting these biological effects?
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
What is metabolomics?
Metabolome can be influenced by:• Genetic factors
• Environmental factors
The metabolome is ever-changing!
Genomics(genome)
Transcriptomics(transcriptome)
Proteomics(proteome)
Metabolomics(metabolome)
Amino
acidsLipids Small CHO
Nucleic
acids
Primary
metabolites
Secondary
metabolitesHormones
Phenotype!
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Untargeted metabolomics vs. targeted analyses
MetabolomicsTargeted analyses
1 to ~dozens of analytes
Work on the front end
Quantitative
Hypothesis driven
Untargeted
metabolomicsTargeted analysis
100s to 1,000s of analytes
Work on the back end
Comparative
Hypothesis generating
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
ProteomicsRe
sp
on
se
GenomicsRe
sp
on
se
MetabolomicsRe
sp
on
se
Adapted from David Wishart’s Canadian Bioinformatics Workshop (www.bioinformatics.ca)
Metabolomics is time-sensitive!
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Metabolomics workflow
Choose
your
samples
Prepare
samples for
analysis
Acquire raw
data
Ask a
scientific
question!Spectral
processing
Post-
acquisition
data
analysis
Metabolite
ID and
annotation
Context-
ualization
Validation
with
targeted
analyses
Integrate with other
omics and meta-data
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Differences between metabolomics and genomics
• Metabolomics has:• Increased diversity in analytes
• Lack of a ‘reference’
• Accurate mass match does not mean an identification
• Larger differences due to instrumentation platforms
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Metabolomics allows global investigation of murine skin
AIN-93G (control)
AIN-93G + 10% tangerine tomato powder
AIN-93G + 10% red tomato powder4 week old,
male SKH1
mice
Week 0 Week 35
sacrifice
Metabolomic
analysisCooperstone et al., Sci Reports, 2017;7:5106
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Metabolomics workflow
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Cooperstone et al., Sci Reports, 2017;7:5106
aIdentities assigned via accurate mass, relative retention time and MS/MS fragmentation patterns.bTomatidine identity confirmed by authentic standard.
Metabolites in murine skin that differentiate animals on control vs. tomato diets linked to tomato glycoalkaloids
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Cholesterol derived, steroidal, nitrogen containing
Found only in the tomato clade
Protective against Fusarium1,2
Bioactivity in vitroColon3, liver3 and prostate4 cancer cell lines
Bioactivity in vivoInhibit skeletal muscle atrophy5, reduce plasma cholesterol6
1Gottleib D Phytopathology 1943;33:1111.2Irving GW et al., Science 1945;102:9–11.
3Lee K-R et al., J. Agric. Food Chem. 2004;52:2832–2839.4Choi SH et al., J Agric. Food Chem. 2012;60:3891–3899.
Tomato Genetics Resource Center
UC Davis, LA2213
5Dyle MC et al., J. Biol. Chem. 2014;289:14913–14924.6Friedman M et al., Food Chem. Toxicol. 2000;38: 549–553.
What are tomato glycoalkaloids?
Solanum pimpinellifolium
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
α-tomatine
tomatidine
hydrolysis &
absorption
Unpublished data
Confirming tomatidine in human plasma
Presence of tomatidine confirmed in human plasma of one individual
consuming tomato juice daily. The top panel is tomatidine standard and the
bottom panel is in human plasma. Both mass chromatograms are extracted at
416.3517 m/z as the [M+H] in positive ion mode.
Human
plasma
Tomatidine
authentic
standard
Tomato glycoalkaloids have plausible biological activity
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Michael Dzakovich
1Blanca et al., BMC Genomics 2015;16:257
Understanding phenotypic diversity in tomato alkaloids
108 accessions, previously genotyped using Tomato Infinium Array (7,720 SNPs)1
2 locations
2 ripeness stagesMature green
Ripe red
684 samples
UPLC-MS/MS targeted alkaloid screening
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Glycoalkaloids in unripe and ripe tomatoes
Mass chromatograms of glycoalkaloids found in mature green (top) and ripe red tomatoes (bottom). Profiles of glycoalkaloids are visually different at these two ripeness stages.
1: dehydrotomatidine, 2: alpha-tomatine, 3: lycoperoside H, hydroxy-alpha-tomatine or pimpifolidine lycotetraose, 4: dihydroxy-alpha-tomatine or hydroxy-pimpifolidine lycotetraose, 5:
lycoperoside A, B or C, 6: esculeoside B and isomers, 7: dehydrolycoperoside F, G or dehydroesculeoside A, 8: lycoperoside F, G or esculeoside A.
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
From bitter to better: Exploring natural variation, bioavailability and tissue distribution of tomato alkaloids
Survey tomato
accessions for
glycoalkaloids
Determine
pharmacokinetics of
glycoalkaloids in humans
Tissue distribution
of tomato
glycoalkaloids/alkaloids
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Looking forward…
• Tomato consumption has the ability to affect cancer development in animals, and the reason why is unclear.
• Metabolomics is a useful technique to comprehensively profile and generate hypotheses as to other putative bioactive compounds from plant foods.
• Tomato glycoalkaloids warrant further investigation into their bioactivty.
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
BIG thanks!+ Troy, Jin, Eduardo, Sean and the Francis lab
Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab
Also BIG thanks
Center for Advanced
Processing and
Packaging Studies
Michael “Tomato” Dzakovich, M.S.Matt Teegarden, Ph.D.
David Francis, Ph.D.
Steven Schwartz, Ph.D.Robin Ralston, M.S., R.D.Ralf Schweiggert, Ph.D.
Earl Harrison, Ph.D.Rachel Kopec, Ph.D.
Tatiana Oberyszyn, Ph.D.Kathy Tober, Ph.D.
Steven Clinton, M.D., Ph.D.Beth Grainger, Ph.D., R.D.Nancy Moran, Ph.D.Jennifer Thomas-Ahner, Ph.D.Remy Powell
OSU’s Foods for Health
Discovery Theme
Initiative
Gregory Lesinski, Ph.D.
Tom Mace, Ph.D.
Janet Novotny, Ph.D.
Itkin et al., Plant Cell 2011
Figure 1. General structure of alkaloids/glycoalkaloids and alkaloids found in tomatoes. Glycoalkaloids refer to
the structures that include sugars, while alkaloids encompass compounds both with and without attached sugars.
The structure on the left as shown is tomatidine. α-Tomatine exists when the lycotetraose (right), is connected to
tomatidine through an O-glycosylation at C-3. Dehydro-products are produced when there is a C5-C6 double
bond. Hydroxy-products are hydroxylated presumably at a number of different locations. Lycoperosides A-C
result from acetylation on the nitrogen-containing ring, while lycoperosides F, G and esculeoside A are
additionally O-glucosylated lycoperosides A, B and C, respectively. Lycoperoside H is a rearrangement and
oxygenation of the nitrogen containing ring from α-tomatine while esculeoside B is glucosylated (at C27)
lycoperoside H.