tomatoes, health & the metabolome

Tomatoes, Health & the Metabolome

Jessica Cooperstone, Ph.D.

Assistant Professor

Horticulture & Crop Science

Food Science & Technology

[email protected]

mailto:[email protected]

Tomato Breeders Roundtable Meeting, April 6, 2018 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. @CooperstoneLab

Are tomatoes healthy?


Better understand the relationship between phytochemicals in plants and health outcomes,

using metabolomics to elucidate these relationships


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)


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


0

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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


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


Cooperstone et al., Food Chem 2016;210:466-472

Carotenoid physical storage form affects processing stability

tetra-cis-lycopene


Carotenoids are higher in human plasma after consuming tangerine tomato juice

E

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D

O

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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

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Control (no juice)

Tangerine tomato juice (2 cans/day, ~325 mL)

Red tomato juice(2 cans/day, ~325 mL)

S

U

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G

E

R

Y

3-5 weeks

n=15

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n=15

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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


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


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500

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3500n

mo

l/L

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Red

Plasma carotenoids after 18 weeks of consumption

of 10% tomato powder containing AIN-93G diet

0%

10%

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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


0.0

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ou

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Male Control

Male Tangerine

Male Red

Cooperstone et al., Sci Reports, 2017;7:5106

0

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Tangerine Red

pm

ol ly

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pe

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/g s

kin

*

Tomatoes can modulate risk for skin cancer as well

*


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?


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!


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


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!


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


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


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


Metabolomics workflow


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


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


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


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


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.


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


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.


BIG thanks!+ Troy, Jin, Eduardo, Sean and the Francis lab


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.

tomatoes, health & the metabolome

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