Tomatoes, Health & the Metabolome

Transcription

1 Tomatoes, Health & the Metabolome Jessica Cooperstone, Ph.D. Assistant Professor Horticulture & Crop Science Food Science & Technology

2 Are tomatoes healthy?

3 Better understand the relationship between phytochemicals in plants and health outcomes, using metabolomics to elucidate these relationships

4 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)

5 Absorbance tetra-cislycopene all-translycopene Nanometers all-trans-lycopene tetra-cis-lycopene

6 Baseline corrected total lycopene (nmol/l TRL ± SEM) Lycopene is more bioavailable from tangerine tomatoes Tangerine tomato Red tomato Hours post meal consumption 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% Tangerine tomato Red tomato cis-lycopene all-trans-lycopene Cooperstone et al., Mol Nutr Food Res 2015:59,

7 Carotenoid physical storage form differs between red and tangerine tomatoes Light micrographs of red and tangerine at 400x magnification. Cooperstone et al., Mol Nutr Food Res 2015:59,

8 Carotenoid physical storage form affects processing stability tetra-cis-lycopene Cooperstone et al., Food Chem 2016;210:

9 Carotenoids are higher in human n=15 plasma after L Tangerine tomato juice O L (2 cans/day, ~325 ml) M consuming tangerine M tomato juice I E Z n=15 E N R O N T R A N D E n=15 Control (no juice) Red tomato juice (2 cans/day, ~325 ml) S U R G E R Y E N R O L L M E N T R A N D O M I Z E n=15 n=15 n=15 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 nmol/l lycopene weeks ab ab bc b c d Control Red Tangerine 3-5 weeks 0 Visit 1 (baseline) Visit 2 (surgery) Different letters denote statistically significant differences via two-way ANOVA with Tukey s posthoc Unpublished test (P<0.05). dat Different letters denote statistically significant differences via two-way ANOVA with Tukey s posthoc test (P<0.05). Unpublished data, Pelotonia IRP

10 Can tomatoes modulate prostate cancer in vivo? Tangerine AIN-93G (control) 4 week old, male TRAMP +/- mice AIN-93G + 10% tangerine tomato powder AIN-93G + 10% red tomato powder Red Week 0 Week 18 sacrifice Carotenoid analysis Tumor pathology tetra-cislycopene all-translycopene Unpublished data, AICR grant #318555

11 nmol/l Plasma carotenoids after 18 weeks of consumption of 10% tomato powder containing AIN-93G diet Red Tangerine and tangerine tomatoes tomatoes lead higher reduce prostate plasma cancer carotenoids by ~40% Tangerine Red 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% High Grade Pin Microscopic Cancer Macroscopic Cancer Control Red Tangerine Most severe cancer pathology observed in each animal, displayed as a percentage. Unpublished data, AICR grant #318555

12 20 Weeks 21 Weeks 22 Weeks 23 Weeks 24 Weeks 25 Weeks 26 Weeks 27 Weeks 28 Weeks 29 Weeks 30 Weeks 31 Weeks 32 Weeks 33 Weeks 34 Weeks 35 Weeks pmol lycopene/g skin Average number of tumors per male mouse Tomatoes can modulate risk for skin cancer as well * Male Control Male Tangerine Male Red * Tangerine Red 0.0 Cooperstone et al., Sci Reports, 2017;7:5106

13 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?

14 What is metabolomics? Genomics (genome) Transcriptomics (transcriptome) Proteomics (proteome) Metabolomics (metabolome) Phenotype! Metabolome can be influenced by: Genetic factors Environmental factors The metabolome is everchanging! Hormones Amino acids Nucleic acids Lipids Primary metabolites Small CHO Secondary metabolites

15 Untargeted metabolomics vs. targeted analyses Targeted analyses 1 to ~dozens of analytes Work on the front end Quantitative Hypothesis driven Metabolomics 100s to 1,000s of analytes Work on the back end Comparative Hypothesis generating Untargeted metabolomics Targeted analysis

16 Response Response Response Metabolomics is time-sensitive! Genomics Proteomics Metabolomics Adapted from David Wishart s Canadian Bioinformatics Workshop (

17 Ask a scientific question! Metabolomics workflow Prepare samples for analysis Acquire raw data Spectral processing Integrate with other omics and meta-data Choose your samples Postacquisition data analysis Validation with targeted analyses Contextualization Metabolite ID and annotation

18 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

19 Metabolomics allows global investigation of murine skin AIN-93G (control) AIN-93G + 10% tangerine tomato powder 4 week old, male SKH1 mice AIN-93G + 10% red tomato powder Week 0 Week 35 sacrifice Cooperstone et al., Sci Reports, 2017;7:5106 Metabolomic analysis

20 Metabolomics workflow

21 Metabolites in murine skin that differentiate animals on control vs. tomato diets linked to tomato glycoalkaloids a Identities assigned via accurate mass, relative retention time and MS/MS fragmentation patterns. b Tomatidine identity confirmed by authentic standard. Cooperstone et al., Sci Reports, 2017;7:5106

22 What are tomato glycoalkaloids? Cholesterol derived, steroidal, nitrogen containing Found only in the tomato clade Protective against Fusarium 1,2 Bioactivity in vitro Colon 3, liver 3 and prostate 4 cancer cell lines Bioactivity in vivo Inhibit skeletal muscle atrophy 5, reduce plasma cholesterol 6 Tomato Genetics Resource Center UC Davis, LA2213 Solanum pimpinellifolium 1 Gottleib D Phytopathology 1943;33: Irving GW et al., Science 1945;102: Lee K-R et al., J. Agric. Food Chem. 2004;52: Choi SH et al., J Agric. Food Chem. 2012;60: Dyle MC et al., J. Biol. Chem. 2014;289: Friedman M et al., Food Chem. Toxicol. 2000;38:

23 Tomato Confirming glycoalkaloids tomatidine have in plausible human plasma biological activity α-tomatine Tomatidine authentic standard hydrolysis & absorption Human plasma Unpublished data tomatidine 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 m/z as the [M+H] in positive ion mode.

24 Understanding phenotypic diversity in tomato alkaloids 108 accessions, previously genotyped using Tomato Infinium Array (7,720 SNPs) 1 2 locations 2 ripeness stages Mature green Ripe red 684 samples UPLC-MS/MS targeted alkaloid screening Michael Dzakovich 1 Blanca et al., BMC Genomics 2015;16:257

25 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.

26 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

27 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.

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

29 Also BIG thanks 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 Gregory Lesinski, Ph.D. Tom Mace, Ph.D. Janet Novotny, Ph.D. OSU s Foods for Health Discovery Theme Initiative Center for Advanced Processing and Packaging Studies

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31 Itkin et al., Plant Cell 2011

32 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.