Diet, the Gut Microbiome and Cancer

Johanna W. Lampe, PhD, RD Fred Hutchinson Cancer Research Center,

Seattle, WA jlampe@fredhutch.org

AICR Conference – Split Session A Dietary Modulation of the Microbiome and Cancer Risk

AICR Conference on Nutrition, Physical Activity, Obesity and Cancer November 14 – 16, 2016 • Bethesda North Marriott • North Bethesda, MD

Dietary Exposures and Cellular Processes Linked to Cancer

Diet

Differentiation

DNA repair

Carcinogen metabolism

Hormonal regulation

Inflammation Immune function

Apoptosis

Proliferation

Adapted from WCRF/AICR 2007 Expert Report

Diet and microbial metabolism

Involvement of Microbial Metabolism

Outline

How does diet affect gut microbial community? How does the gut microbiome affect components

of diet?

Gut Microbial Metabolism Obtain energy and nutrients to

live and reproduce

Microbiome: >100 times as many genes as human genome

Carry out reactions that human gut enzymes cannot Fermentation Denitrification Sulfate reduction Aromatic fission Hydrolysis/deconjugation

The indigestibles The leftovers

Food

Human digestion

Bacterial metabolism

Fermentation of Carbohydrates: Sugars, Dietary Fiber and Resistant Starch

Acetate Propionate Butyrate

>81 different glycoside hydrolase families

Gill et al, Science, 2006 Tremaroli & Bäckhed, Nature, 2012

Microbial Metabolism of Proteins & Amino Acids

Aromatic Amino acids

Other Amino acids

Sulfur Amino acids

Sulfur compounds

Phenols and indoles

Ammonia NH3

+/NH4

Amines H2, CO2, CH4 Organic acids

Proteins Peptides hydrolysis

α, β elimination deamination deamination &

fermentation decarboxylation

Adapted from Nyangale et al. J Proteome Res, 2012

Gut Microbiome and Dietary Fats

Joyce and Gahan, Ann Rev Food Sci Technol, 7:313, 2016

Metabolism of lipids Bile acid metabolism Primary to

secondary bile acids

Altered microbial community

Changes in signaling to liver

How does diet affect the gut microbiome? Evidence from:

Observational studies Globally distinct populations Long-term food pattern consumption

Short-term dietary interventions Low- vs high-fiber diets Animal vs plant food sources Macronutrient ratios

Reviewed in Arora & Bäckhed. J Int Med 280:339, 2016

Global Population Differences: Children in Rural Africa (BF) vs Urban Europe (EU)

De Filippo et al PNAS 107:14691, 2010

Global Population Differences: African-Americans in US vs Native South Africans

Ou et al, Am J Clin Nutr 98:111, 2013

Microbiota Change with Diet Switch in African Americans and Native South Africans

2-wk feeding study

US AAs fed: Fiber: 14→55g/d Fat: 35→16%

Native Africans fed: Fiber: 66→12g/d Fat: 16→52%

African Americans

Native Africans

Afric

an d

iet

Wes

tern

die

t

Wes

tern

die

t

Afric

an d

iet

O’Keefe et al, Nat Comm 6:6342, 2015

Diet Pattern Change and Gut Microbiome : Fat & Fiber and Colorectal Cancer Risk Factors

High-fiber, low-fat diet changed microbiome: Increased saccharolytic fermentation and

butyrate production Decreased secondary bile acid synthesis

Functional changes in gut microbiota were accompanied by colorectal cancer relevant changes in colonic mucosal proliferation and inflammation

O’Keefe et al, Nat Comm 6:6342, 2015

Short-Term Feeding of Plant- and Animal-Based Diets Alters Gut Microbiota

10 subjects tracked across plant- and animal-based diet treatments. Animal-based diet increased bile-

tolerant microorganisms and decreased microbes that metabolize plant polysaccharides. Bacterial metabolic gene

expression (RNA-seq) tends to cluster by diet.

Diet doesn’t always overcome inter-individual differences in GMC structure (16S rRNA).

12 David et al, Nature, 505:559 2014

Diet Patterns and Gut Microbiome: Controlled Low and High-Glycemic Load Diets

4-week randomized crossover feeding study in 12 healthy U.S. men and women.

Eucaloric diets with same macronutrient distribution, except: • Low GL: 55 g fiber • High GL: 28 g fiber

Discriminant analysis of microbiome distinguishes by diet

Neuhouser et al, J Nutr 2012

Hullar & Lampe, unpublished data, 2016

Martens E. Nature 529:158-159, 2016 Sonnenburg, E. et al. Nature 529: 212-215, 2016

Multigenerational Effects of Diet on Bacterial Diversity

How does the gut microbiome affect diet?

Alters exposure to nutrients and bioactives Generates new compounds, which: Serve as energy source Regulate metabolism Reduce inflammation Cause oxidative stress

Bacteria Can Produce New Compounds from Food Components

Food Component Bacterial Metabolite Dietary fiber Butyrate and other SCFAs Choline Trimethylamine Soy isoflavones Equol, O-desmethylangolensin Plant lignans Enterodiol, enterolactone Ellagitannins Urolithins A and B Anthocyanins Hippuric acid & small phenolics Glucosinolates Isothiocyanates Linoleic Acid Conjugated linoleic acid

Diet-Microbial Community Interaction Affects Exposure to Dietary Metabolites

Russell et al., Am J Clin Nutr 2011;93:1062–72.

Tota

l N-n

itros

o co

mpo

unds

(ng/

ml)

a

c

b

a,b,c significantly different p<0.001

17 obese men, randomized cross-over design

4 weeks of weight-reduction diets: high-protein; med carb high-protein; low-carb

HPLC diet resulted in decrease

in fecal cancer-protective metabolites (butyrate) and increased concentrations of hazardous metabolites (N-nitroso compounds).

Protein, % 13 28 29

Fat, % 37 37 66

Carb, % 50 35 5

NSP, g 22 9 13

Choline Metabolism, TMAO, and Cancer?

Shared risk factors with atherosclerosis: Oxidative stress

Inflammation

Insulin resistance

TMAO and aggressive prostate cancer in ATBC: OR: 1.36 (1.02–1.81), p=0.039

Choline metabolites and colorectal cancer in WHI: OR: 1.65 (1.17-2.34)

Tang et al. NEJM, 2013

Dietary phosphatidyl

choline

Choline

Trimethylamine

Trimethylamine N-oxide

Betaine

Atherosclerosis

Death Stroke Heart attack

Gut microbiota

Mondul et al. Int J Cancer, 2015 Bae et al, Cancer Res, 2014

Anthocyanin Metabolism by Gut Microbes

Higher consumption of high-anthocyanin foods inversely associated with risk of hypertension and CVD mortality.

Low bioavailability of parent compounds.

Microbial hydrolysis of glucosides and ring cleavage yield range of small phenolics detected in serum and urine.

De Ferrars et al, Br J Pharmacol, 2014

Metabolic Phenotypes: Microbial Production of Equol and ODMA From Soy Isoflavone Daidzein

O

O

HO

OH

OHO

OH

O

O

HO

OH

OH

O

HO

OH

OHO

OHOH

Daidzein Dihydrodaidzein

EquolO-Desmethylangolensin

Cis/Trans-isoflavan-4-ol

20-60% of individuals produce

80-90% of individuals produce

Daidzein-Metabolizing Phenotypes and Cardiovascular Risk among 595 Chinese Postmenopausal Women

Equol producers, compared to nonproducers, had: Higher fat-free mass

Lower systolic and diastolic blood pressure

Lower serum triglyceride), hs-CRP and FFA

O-DMA producers, compared to nonproducers, had: Lower BMI and %body fat

Lower total cholesterol

Habitual soy isoflavone intake had little relation to CVD risk factors in either metabolizing phenotype.

Liu Z-m, PLoS ONE 9(2): e87861, 2014.

Obese Adults More Likely to be ODMA-Nonproducer Phenotype

Frankenfeld et al., Eur J Clin Nutr, 2014

18 to <25 kg/m2

25 to 29.9 kg/m2

30+ kg/m2 P-trend

ODMA producers n (%) 142 (59.9) 71 (30.0) 24 (10.1) ODMA nonproducers 29 (48.3) 17 (28.3) 14 (23.3) OR* REF 1.0 (0.5, 2.1) 2.8 (1.2, 6.2) 0.032

Equol producers n (%) 77 (62.1) 32 (25.8) 15 (12.1) Equol nonproducers 94 (54.3) 56 (32.4) 23 (13.3) OR* REF 1.3 (0.7, 2.2) 1.1 (0.5, 2.2) 0.629

*n=297; adjusted for age (in years), race, and gender and menopausal status.

LEGEND: 1. O-deglycosylation 2. O-demethylation 3. dehydrogenation 4. dehydroxylation

Enterodiol (END) Enterolactone (ENL)

Secoisolariciresinol Diglucoside

Secoisolariciresinol)

1

2

4

3

4 3

Enterolignan Production by Gut Bacteria

Gut Microbial Diversity Differs by Tertiles of Urinary Enterolactone Excretion

05

10152025

no ENL Low ENL High ENL

Dive

rsity

Dive

rsity

b b

Low ENL (T1)

Medium ENL (T2)

High ENL (T3)

a

Hullar et al, CEBP, 2015

Gut Microbiome Associated with Urinary Enterolactone (ENL) Excretion

GMC composition is significantly different between high and low ENL excreters (MRPP, p<0.0005).

Association remains significant with adjustment for fiber intake and adiposity.

Low ENL clusters together

Hullar et al, CEBP, 2015

Low ENL

High ENL

Phylogenetic Distribution of Microbiome in High-ENL Excreters

Unique genera high ENL excreters

Distributed across Phyla

Firmicutes

Summary Diet can change the gut microbiome and its impact on

the host. Is it the microbe itself or a consortia of microbes? Is it the metabolites produced by microbes? Or the combination of both?

Availability of nutrients or bioactive substances important for health can be influenced by gut microbiota.

Metabolic phenotypes integrate diet and microbial action.

Better understanding the impact of the bacterial metabolites on regulatory pathways may help guide future diet and cancer prevention strategies.

Supported by: US National Cancer Institute Kellogg Corporate Citizens Fund Fred Hutch

J Lampe Lab

* Meredith Hullar Isaac Elkon Lisa Levy Fei Li Sandi Navarro Wendy Thomas Elizabeth Traylor Seth Yoder

Texas A&M University Robert Chapkin Ivan Ivanov

Hawaii Cancer Center Loic Le Marchand

University of Bristol Charlotte Atkinson

University of Helsinki Kristiina Wähälä

FHCRC and UW collaborators Mario Kratz Marian Neuhouser Tim Randolph Ali Shojae George Mason U Cara Frankenfeld