Gegen Qinlian decoction enhances the effect of PD-1 blockade in colorectal cancer with microsatellite stability by remodelling the gut microbiota and the tumour microenvironment

From BugSigDB
Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18
study design
Citation
PMID PubMed identifier for scientific articles.
DOI Digital object identifier for electronic documents.
URI
Authors
Lv J, Jia Y, Li J, Kuai W, Li Y, Guo F, Xu X, Zhao Z, Lv J, Li Z
Journal
Cell death & disease
Year
2019
Therapeutic antibodies targeting PD-1 have made major breakthroughs in cancer treatment. However, the majority of colorectal cancer (CRC) cases are microsatellite stable (MSS) and do not respond to anti-PD-1-based immunotherapy. Combination therapy will be an ideal strategy to overcome this limitation. Gegen Qinlian decoction (GQD), a classical traditional Chinese medicine (TCM) formula, has been clinically proven to be effective in the treatment of ulcerative colitis (UC) and type 2 diabetes mellitus. Here, a systemic pharmacological study revealed that GQD acts through multiple targets and pathways in the human body. Combination therapy with GQD and anti-mouse PD-1 potently inhibited the growth of CT26 tumours in a xenograft model. Gut microbiota analysis revealed that combination therapy with GQD and anti-mouse PD-1 significantly enriched for s__Bacteroides_acidifaciens and s__uncultured_organism_g__norank_f__Bacteroidales_S24-7_group. Based on metabolomic analyses, profoundly altered metabolites were identified in the combination therapy group. Two metabolic signalling pathways, namely, glycerophospholipid metabolism and sphingolipid metabolism, were explored. In particular, we found that combination therapy with GQD and anti-mouse PD-1 significantly increased the proportion of CD8+ T cells in peripheral blood and tumour tissues. Direct treatment with GQD and anti-mouse PD-1 increased the expression of IFN-γ, which is a critical factor in antitumour immunotherapy. In addition, combination therapy with GQD and anti-mouse PD-1 downregulated PD-1 and increased IL-2 levels, suggesting that the combination therapy could effectively restore T-cell functions by suppressing inhibitory checkpoints. The application of the Chinese medicinal formula GQD with PD-1 blockade-based immunotherapy can be a novel therapeutic strategy for CRC patients with MSS tumours.

Experiment 1


Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1, WikiWorks, Peace Sandy

Subjects

Location of subjects
China
Host species Species from which microbiome was sampled. Contact us to have more species added.
Mus musculus
Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology
Feces Cow dung,Cow pat,Droppings,Dung,Excrement,Excreta,Faeces,Fecal material,Fecal matter,Fewmet,Frass,Guano,Matières fécales@fr,Merde@fr,Ordure,Partie de la merde@fr,Piece of shit,Porción de mierda@es,Portion of dung,Portion of excrement,Portion of faeces,Portion of fecal material,Portion of fecal matter,Portion of feces,Portion of guano,Portion of scat,Portionem cacas,Scat,Spoor,Spraint,Stool,Teil der fäkalien@de,Feces,feces
Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology
Colorectal cancer cancer of colorectum,cancer of large bowel,cancer of large intestine,cancer of the large bowel,colon cancer,colorectal cancer,colorectum cancer,CRC,large intestine cancer,malignant colorectal neoplasm,malignant colorectal tumor,malignant colorectum neoplasm,malignant large bowel neoplasm,malignant large bowel tumor,malignant large intestine neoplasm,malignant large intestine tumor,malignant neoplasm of colorectum,malignant neoplasm of large bowel,malignant neoplasm of large intestine,malignant neoplasm of the large bowel,malignant neoplasm of the large intestine,malignant tumor of large bowel,malignant tumor of large intestine,malignant tumor of the large bowel,malignant tumor of the large intestine,Colorectal cancer
Group 0 name Corresponds to the control (unexposed) group for case-control studies
non-GQD
Group 1 name Corresponds to the case (exposed) group for case-control studies
low-dose GQD (300 mg/kg)
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
BALB/c mice (male, ~20 g, aged 5 weeks) were fed adaptively for 1 week under specific pathogen-free conditions with food and water provided ad libitum. Then the mice were orally gavaged with 300 mg/kg Gegen Qinlian decoction (GQD), a classical traditional Chinese medicine (TCM) formula, once a day for 10 days. The mouse colorectal carcinoma cell line CT26 (~2.5 × 106 cells/mouse) were transplanted subcutaneously into the left axillary region of each mouse and allowed ~1 week to establish tumours.
Group 0 sample size Number of subjects in the control (unexposed) group
20
Group 1 sample size Number of subjects in the case (exposed) group
23

Lab analysis

Sequencing type
16S
16S variable region One or more hypervariable region(s) of the bacterial 16S gene
V4
Sequencing platform Manufacturer and experimental platform used for quantifying microbial abundance
Illumina

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).
relative abundances
Statistical test
Mann-Whitney (Wilcoxon)
Significance threshold p-value or FDR threshold used for differential abundance testing (if any)
0.05
MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?
Yes

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness
unchanged
Chao1 Abundance-based estimator of species richness
unchanged

Signature 1

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: Figure 3c

Description: Differential microbial abundance between the GQD (300 mg/kg) vs. the non-GQD groups based on the Wilcoxon rank-sum test.

Abundance in Group 1: increased abundance in low-dose GQD (300 mg/kg)

NCBI Quality ControlLinks
Candidatus Stoquefichus
Clostridiales bacterium CIEAF 012
Lactobacillus
Proteus mirabilis
[Clostridium] innocuum
unclassified Erysipelotrichaceae
Parasutterella
Roseburia
Candidatus Saccharimonas

Revision editor(s): Mary Bearkland, Claregrieve1

Signature 2

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: Figure 3c

Description: Differential microbial abundance between the GQD (300 mg/kg) vs. the non-GQD groups based on the Wilcoxon rank-sum test.

Abundance in Group 1: decreased abundance in low-dose GQD (300 mg/kg)

NCBI Quality ControlLinks
Candidatus Saccharimonas
Peptococcaceae
Staphylococcus sp.
Turicibacter
Tyzzerella
uncultured Oscillibacter sp.

Revision editor(s): Mary Bearkland, Claregrieve1

Experiment 2


Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1, WikiWorks, Victoria

Differences from previous experiment shown

Subjects

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
BALB/c mice (male, ~20 g, aged 5 weeks) were fed adaptively for 1 week under specific pathogen-free conditions with food and water provided ad libitum. Then the mice were orally lavaged with 300 mg/kg Gegen Qinlian decoction (GQD), a classical traditional Chinese medicine (TCM) formula, once a day for 10 days. The mouse colorectal carcinoma cell line CT26 (~2.5 × 106 cells/mouse) were transplanted subcutaneously into the left axillary region of each mouse and allowed ~1 week to establish tumours.

Lab analysis

Statistical Analysis

Statistical test
LEfSe
LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool
2

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness
unchanged
Chao1 Abundance-based estimator of species richness
unchanged

Signature 1

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: Figure 3d

Description: Differential microbial abundance between the GQD (300 mg/kg) vs. the non-GQD groups based on LefSe analysis (LDA > 2)

Abundance in Group 1: increased abundance in low-dose GQD (300 mg/kg)

NCBI Quality ControlLinks
Candidatus Stoquefichus
Clostridiales bacterium CIEAF 012
Erysipelotrichaceae
Erysipelotrichales
Erysipelotrichia
Ligilactobacillus murinus
Proteus
Proteus mirabilis
[Clostridium] innocuum
unclassified Erysipelotrichaceae
unclassified Parasutterella
uncultured Roseburia sp.

Revision editor(s): Mary Bearkland, Claregrieve1

Signature 2

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/04

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: Figure 3D

Description: Differential microbial abundance between the GQD (300 mg/kg) vs. the non-GQD groups based on LefSe analysis (LDA > 2)

Abundance in Group 1: decreased abundance in low-dose GQD (300 mg/kg)

NCBI Quality ControlLinks
uncultured Bacteroidales bacterium
Gammaproteobacteria
Enterobacterales
Enterobacteriaceae
Peptococcaceae
Bacillales
Staphylococcus
Candidatus Saccharimonas
Candidatus Saccharibacteria
Turicibacter
Tyzzerella
Oscillibacter sp.

Revision editor(s): Mary Bearkland, Claregrieve1

Experiment 3


Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/13

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1, WikiWorks, Victoria

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies
control
Group 1 name Corresponds to the case (exposed) group for case-control studies
low-dose GQD (300 mg/kg) + PD-1
Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group
BALB/c mice (male, ~20 g, aged 5 weeks) were fed adaptively for 1 week under specific pathogen-free conditions with food and water provided ad libitum. Then the mice were orally gavaged with 300 mg/kg Gegen Qinlian decoction (GQD), a classical traditional Chinese medicine (TCM) formula, once a day for 10 days. The mouse colorectal carcinoma cell line CT26 (~2.5 × 106 cells/mouse) were transplanted subcutaneously into the left axillary region of each mouse and allowed ~1 week to establish tumours. When the tumours reached a size of 50mm3, the mice were intraperitoneally (i.p.) injected with 250 μg of anti-mouse PD-1 mAb. The mice in the control group were administered the same volume of PBS. All mice were injected five times at 3-day intervals with antimouse PD-1 mAb or PBS
Group 0 sample size Number of subjects in the control (unexposed) group
11
Group 1 sample size Number of subjects in the case (exposed) group
12

Lab analysis

Statistical Analysis

Statistical test
Mann-Whitney (Wilcoxon)
LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool
Not specified

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness
unchanged
Chao1 Abundance-based estimator of species richness
unchanged

Signature 1

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/13

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: Figure 4f

Description: Differential microbial abundance between the GQD (300 mg/kg) + PD-1 group vs. the control group based on the Wilcoxon rank-sum test.

Abundance in Group 1: increased abundance in low-dose GQD (300 mg/kg) + PD-1

NCBI Quality ControlLinks
Alistipes
Bacteroidales
Bacteroides acidifaciens
Candidatus Stoquefichus
Clostridiales bacterium
Lachnospiraceae bacterium
Peptococcaceae
Prevotellaceae
Ruminiclostridium
uncultured Eubacterium sp.
uncultured Oscillospiraceae bacterium
Eubacterium xylanophilum
unclassified Mollicutes

Revision editor(s): Mary Bearkland, Claregrieve1

Signature 2

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/14

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: Fig 4f

Description: Differential microbial abundance between GQD(300mg/kg)+PD-1 group and control group

Abundance in Group 1: decreased abundance in low-dose GQD (300 mg/kg) + PD-1

NCBI Quality ControlLinks
Anaerotruncus sp.
Bifidobacterium animalis
Candidatus Saccharimonas
Roseburia sp.
unclassified Lachnospiraceae
uncultured Bacteroidales bacterium
uncultured Candidatus Saccharimonas sp.
uncultured Coprococcus sp.
uncultured Eubacteriales bacterium
uncultured Ruminiclostridium sp.
Oscillospiraceae bacterium
Clostridiales bacterium

Revision editor(s): Mary Bearkland, Claregrieve1

Experiment 4


Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/15

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1, WikiWorks, Victoria

Differences from previous experiment shown

Subjects

Group 1 name Corresponds to the case (exposed) group for case-control studies
low-dose GQD (300 mg/kg)

Lab analysis

Statistical Analysis

Statistical test
LEfSe

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness
increased
Chao1 Abundance-based estimator of species richness
increased

Signature 1

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/18

Curated date: 2022/09/15

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: Supp fig S6D

Description: Differential microbial abundance between GQD(300mg/kg) treated mice and controls

Abundance in Group 1: increased abundance in low-dose GQD (300 mg/kg)

NCBI Quality ControlLinks
Bacteroidaceae
Bacteroides
Eubacterium xylanophilum
Hungatella
Parabacteroides
Porphyromonadaceae
unclassified Coriobacteriaceae

Revision editor(s): Mary Bearkland, Claregrieve1

Signature 2

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/19

Curated date: 2022/09/15

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: supp fig S6

Description: Differential microbial abundance between GQD(300mg/kg) treated mice and controls

Abundance in Group 1: decreased abundance in low-dose GQD (300 mg/kg)

NCBI Quality ControlLinks
Actinomycetota
Anaeroplasma
Anaeroplasmataceae
Anaeroplasmatales
Bifidobacteriaceae
Bifidobacteriales
Bifidobacterium
Blautia
Christensenellaceae
Coprococcus
Coriobacteriaceae
Coriobacteriales
Deltaproteobacteria
Desulfovibrio
Desulfovibrionales
Desulfurococcaceae
Enterorhabdus
Tyzzerella
[Eubacterium] brachy
unclassified Bacteroidales
unclassified Christensenellaceae
unclassified Lachnospiraceae

Revision editor(s): Mary Bearkland, Claregrieve1

Experiment 5


Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/20

Curated date: 2022/09/17

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1, WikiWorks, Victoria

Differences from previous experiment shown

Subjects

Group 1 name Corresponds to the case (exposed) group for case-control studies
PD-1 group
Group 1 sample size Number of subjects in the case (exposed) group
9

Lab analysis

Statistical Analysis

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness
unchanged
Chao1 Abundance-based estimator of species richness
unchanged

Signature 1

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/20

Curated date: 2022/09/17

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland

Source: supp fig S6

Description: Supplementary Figure S6: (E) LDA scores computed for differentially abundant taxa in the faecal microbiomes of mice from each group.

Abundance in Group 1: increased abundance in PD-1 group

NCBI Quality ControlLinks
Adlercreutzia
Anaeroplasma
Anaeroplasmataceae
Anaeroplasmatales
Thomasclavelia
Eubacterium ventriosum
Hungatella
Mollicutes
Mycoplasmatota

Revision editor(s): Mary Bearkland

Signature 2

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/20

Curated date: 2022/09/17

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland

Source: supp fig S6

Description: Supplementary Figure S6: (E) LDA scores computed for differentially abundant taxa in the faecal microbiomes of mice from each group.

Abundance in Group 1: decreased abundance in PD-1 group

NCBI Quality ControlLinks
Oscillibacter
Bacillota

Revision editor(s): Mary Bearkland

Experiment 6


Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/20

Curated date: 2022/09/17

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1, WikiWorks, Victoria

Differences from previous experiment shown

Subjects

Group 1 name Corresponds to the case (exposed) group for case-control studies
low dose 300 mg/kg GQD + PD-1 group
Group 1 sample size Number of subjects in the case (exposed) group
12

Lab analysis

Statistical Analysis

Signature 1

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/20

Curated date: 2022/09/17

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: supp fig S6

Description: Differentially abundant taxa between control mice and mice treated with 300mg/kg GQD + PD-1

Abundance in Group 1: increased abundance in low dose 300 mg/kg GQD + PD-1 group

NCBI Quality ControlLinks
Alistipes
Bacteroidaceae
Bacteroides
Candidatus Stoquefichus
Prevotellaceae
Rikenellaceae
Ruminiclostridium
unclassified Prevotellaceae
Oscillospiraceae

Revision editor(s): Mary Bearkland, Claregrieve1

Signature 2

Reviewed Marked as Reviewed by Claregrieve1 on 2022/11/20

Curated date: 2022/09/17

Curator: Mary Bearkland

Revision editor(s): Mary Bearkland, Claregrieve1

Source: Supp fig S6

Description: Differentially abundant taxa between control mice and mice treated with 300mg/kg GQD + PD-1

Abundance in Group 1: decreased abundance in low dose 300 mg/kg GQD + PD-1 group

NCBI Quality ControlLinks
Anaerotruncus
Bacteroidales
Bifidobacteriaceae
Bifidobacteriales
Bifidobacterium
Candidatus Saccharibacteria

Revision editor(s): Mary Bearkland, Claregrieve1