Intestinal Blautia regulates gastrointestinal toxicity in cancer patients receiving standard dose chemotherapy

Wardill HR, Bowen JM, Secombe KR, Tissing WJE, Harmsen HJM, Stringer AM, Al-DasooqiN, Mayo B and Gibson RJ.

Dr Hannah Wardill, PhDNHMRC CJ Martin Biomedical Research FellowThe University of Adelaide (Australia) University Medical Centre Groningen (Netherlands)

@hannahrwardill@ToxicitiesGroup

Conflict of interest Hannah Wardill, PhD

Has no real or apparent conflicts of interest to report.

The microbiome: a new risk prediction tool for

gastrointestinal toxicity?

The microbiome: a new risk prediction tool?• Microbiome is unique and individualized

The microbiome: a new risk prediction tool?• Microbiome is unique and individualized

• Microbiome is uniquely position to modulate cancer treatment efficacy and toxicity due to influence on:

• Drug metabolism • Mucosal immunology / barrier integrity / inflammation • Tolerance / immunogenic cell death

The microbiome: a new risk prediction tool?• Microbiome is unique and individualized

• Microbiome is uniquely position to modulate cancer treatment efficacy and toxicity due to influence on:

• Drug metabolism • Mucosal immunology / barrier integrity / inflammation • Tolerance / immunogenic cell death

• Unlike human genome, the microbiome is highly plastic enabling risk modification and fine tuning of treatment outcomes

Retrospective cohort investigation • Archival fecal samples from N=12 patients undergoing standard dose 5-FU

based chemotherapy for CRC and breast cancer

Retrospective cohort investigation • Archival fecal samples from N=12 patients undergoing standard dose 5-FU

based chemotherapy for CRC and breast cancer

• Patients donated N=2 stool samples samples:

• 1 X before chemotherapy cycle • 1 X day 5 (peak diarrhea)

Retrospective cohort investigation • Archival fecal samples from N=12 patients undergoing standard dose 5-FU

based chemotherapy for CRC and breast cancer

• Patients donated N=2 stool samples samples:

• 1 X before chemotherapy cycle • 1 X day 5 (peak diarrhea)

• Toxicity was assessed using NCI CTCAE v5.0; • Toxic = G3+ diarrhea• Non-toxic = G0/G1 diarrhea

5-FU based chemotherapy disrupts microbial diversity and composition

Figure 1: Species diversity (Shannon’s index) pre- and post-chemotherapy in toxic and non-toxic patients.

T0 T50

1

2

3

4

Sha

nnon

's d

iver

sity

inde

x

Toxic patients Non-toxic patients

Non-Toxic-60

-40

-20

0

20

Cha

nge

in S

peci

es D

ives

ity

(% re

lativ

e to

bas

elin

e)

5-FU based chemotherapy disrupts microbial diversity and composition

Figure 2: Significantly

affected bacterial species in toxic and

non-toxic individuals.

T0 T5-5

0

5

10

15

Rel

ativ

e A

bund

ance

Faecalibacterium prausnitzii

T0 T5-20

5

10

15

20

Rel

ativ

e A

bund

ance

E coli

T0 T5-5

0

5

10

15

20

Rel

ativ

e A

bund

ance

Intestinibacter bartlettii

T0 T5-20

0

20

40

60

80Streptococcus thermophilus

Rel

ativ

e A

bund

ance

T0 T5

0

5

10

Rel

ativ

e A

bund

ance

Ruminococcus bromii

Toxic and non-toxic patients have distinct microbial signatures before chemotherapy

-1.0 -0.5 0.5 1.0

-0.8

-0.6

-0.4

-0.2

0.2

0.4

0.6

0.8

PC2

P

C3

Non-toxic Toxic

16%

11%

Figure 3: Principle component analysis of pre-treatment microbiome composition

Toxic and non-toxic patients have distinct microbial signatures before chemotherapy

TOXIC NON-TOXIC

Figure 4: Microbial composition (genera level) in toxic and non-toxic individuals

Toxic and non-toxic patients have distinct microbial signatures before chemotherapy

TOXIC NON-TOXIC

before chemotherapy

Figure 4: Microbial composition (genera level) in toxic and non-toxic individuals

Figure 5: Relative abundance of Blautia in toxic and non-

toxic individual.

Blautia correlates with toxicity outcome

• Microbial composition aligning with PC considered protective

• Correlated species with PC2 -1.0 -0.5 0.5 1.0

-0.8

-0.6

-0.4

-0.2

0.2

0.4

0.6

0.8

PC2

P

C3

Non-toxic Toxic

16%

11%

Blautia correlates with toxicity outcome

! Microbial composition aligning with PC considered protective

! Correlated species with PC2 -1.0 -0.5 0.5 1.0

-0.8

-0.6

-0.4

-0.2

0.2

0.4

0.6

0.8

PC2

P

C3

Non-toxic Toxic

16%

11%

Collinsellaaerofaciens

Streptococ. Thermophil.

Blautia lutiRuminococcus

lactaris

Correlation coefficient

-0.839*** -0.593** 0.744** 0.616**

What is driving this phenomenon?

Blautia luti in vitro activity• Isolated blautia luti from fresh human faeces (healthy individual)

• Cultured anaerobically in YCFAG, isolated supernatant via centrifugation

• Investigated impact of blautia luti supernatant (B-SPN) on:

• Colonic epithelial proliferation (xCELLigence system)

• Epithelial barrier function (trans-epithelial electrical resistance)

Conclusions• 5-FU based chemotherapy causes microbial dysbiosis reflected by a decrease in

species diversity, a loss of butyrate-producing commensals and an increase in opportunistic pathogens

• Pre-treatment microbial composition critical in determining outcomes

• Blautia genera associated with favourable toxicity outcomes

• Restoration of blautia genera may be important in protecting against gastrointestinal toxicity, via:

• Promotion of epithelial restitution• Restoration of the intestinal barrier • Stimulation of commensal expansion

Acknowledgments Wim Tissing, Hermie Harmsen and Ana Rita da Silva Ferreira: University Medical Centre Groningen (Netherlands)

Kate Secombe, Joanne Bowen, Rachel Gibson, Ysabella Van Sebille, Bronwen Mayo, Noor Al-Dasooqi, Dorothy Keefe and Imogen Ball:The University of Adelaide and University of South Australia (Australia)

FUNDING National Health and Medical Research CommitteeThe University of AdelaideThe Royal Adelaide Hospital

@hannahrwardill@ToxicitiesGroup