Dysbiosis of the duodenal microbiota as a diagnostic marker for pancreaticobiliary cancer

Table 1 Final patient diagnoses

Benign group (n = 22)		Malignant group (n = 12)
Chronic pancreatitis	6	Pancreatic cancer, stage (I/II/III/IV)	9 (2/5/1/1/)
IPMN	5	Biliary ductal cancer, stage (I/II/III/IV)	3 (1/2/0/0)
GB ADM	3
Autoimmune pancreatitis	3
CBD stricture of unknown origin	2
Serous cystic neoplasm	2
CBD stone	1

IPMN: Intraductal papillary mucinous neoplasm; GB ADM: Gallbladder adenomyomatosis; CBD: Common bile duct.

Table 2 Comparison of patient characteristics, tumor markers, and microbiomes

	Benign group (n = 22)	Malignant group (n = 12)	P value
Age, yr (mean ± SD)	63.0 ± 12.2	73.0 ± 8.3	0.016
Sex (male/female)	8/14	4/8	1.0
Reduction in body weight ≥ 5 kg within 6 mo before duodenal juice sampling, n (%)	1 (4.5)	2 (16.7)	0.28
Intake of proton pump inhibitors, n (%)	4 (18.2)	2 (16.7)	1.0
CA19-9, U/mL, median (range)	5.4 (2.0-54.8)	22.8 (2.0-9893.2)	0.03

CA19-9: Cancer antigen 19-9.

Table 3 Microbiome comparison

	Benign group (n = 22)	Malignant group (n = 12)	P value
Bacteroides, %, median (range)	4.3 (0-26.1)	5.6 (0-46.4)	0.55
Bifidobacterium, %, median (range)	0 (0-0.5)	0.3 (0-4.5)	< 0.05
Clostridium cluster IV, %, median (range)	2.9 (0-10.8)	3.4 (0-8.8)	0.80
Clostridium cluster IX, %, median (range)	4.7 (0.6-19.5)	4.9 (0-17.8)	0.68
Clostridium cluster XI, %, median (range)	0 (0-0)	0 (0-0)
Clostridium cluster XVIII, %, median (range)	1.3 (0-3.9)	3.6 (0.8-14.9)	0.006
Clostridium subcluster XIVa, %, median (range)	5.1 (0-23.1)	6.4 (2.9-13.7)	0.38
Lactobacillales, %, mean ± SD	63.0 ± 19.7	62.6 ± 18.3	0.95
Prevotella, %, median (range)	2.2 (0-25.9)	0.1 (0-10.9)	0.04
Others, %, median (range)	4.9 (2.5-20.4)	4.1 (1.5-6.3)	0.14

Table 4 Effects of age on cancer antigen 19-9 levels and the human microbiome

	Age < 69 yr (n = 17)	Age ≥ 69 yr (n = 17)	P value
CA19-9, U/mL, median (range)	7.1 (2-129.3)	4.9 (2-9893.2)	0.77
Bifidobacterium, %, median (range)	0.3 (0-0.5)	0 (0-4.5)	0.3
Clostridium cluster XVIII, %, median (range)	2.6 (0-5.7)	1.8 (0-14.9)	0.82
Prevotella, %, median (range)	1.3 (0-18.2)	1.9 (0-25.9)	0.56

CA19-9: Cancer antigen 19-9.

Table 5 Diagnosis of pancreaticobiliary cancer by the combination of Clostridium cluster XVIII and cancer antigen 19-9 levels

	Cutoff value	Sensitivity	Specificity
CA19-9	18.8 U/mL	66.7% (8/12)	76.2% (16/211)
Clostridium cluster XVIII	3.038%	58.3% (7/12)	95.2% (20/211)
Combination of Clostridium cluster XVIII and CA19-9		91.7% (11/12)	71.4% (15/211)

¹CA19-9 data were missing for a patient in the benign group.

CA19-9: Cancer antigen 19-9.

Table 6 Past reports on microbes and pancreaticobiliary cancer

Disease	Ref.	Microbes	Sample	Role
Pancreatic cancer	Michaud et al[30]	A history of periodontal diseases		Risk factor
	Farrell et al[28]	A combination of Neisseria elongate and Streptococcus mitis	Oral	Distinguishing from healthy controls
	Torres et al[29]	Ratio of Leptotrichia to Porphyromonas	Saliva	Higher in pancreatic cancer patients
	Fan et al[27]	Porphyromonas gingivalis	Oral, antibody	Risk factor
	Olson et al[39]	Firmicutes	Oral	More abundant
	Lu et al[40]	Leptotrichia, Fusobacterium, Rothia, Actinomyces, Corynebacterium, Atopobium, Peptostreptococcus, Catonella, Oribacterium, Filifactor, Campylobacter, Moraxella, Tannerella	Tongue coating	More prevalent
	Mei et al[46]	Acinetobactor, Aquabacterium, Oceanobacillus, Rahnella, Massilia, Delftia, Deinococcus, Sphingobium	Duodenal mucosa	More abundant
	Mitsuhashi et al[43]	Fusobacterium species	Cancer tissue	Poor prognosis
	Riquelme et al[44]	Pseudoxanthomonas, Streptomyces, Saccharopolyspora, Bacillus clausii	Cancer tissue	Long-term survival
Pancreatic and ampullary cancer	Di Calro et al[45]	Escherichia coli, Klebsiella pneumoniae	Bile juice	Predictor for survival
IPMN with high-grade dysplasia	Gaiser et al[47]	Granulicatella adiacens, Fusobacterium nucleatum	Cyst fluid	More abundant

IPMN: Intraductal papillary mucinous neoplasm.

Table 7 Microbiome comparison in patients with pancreatic disease

	Benign pancreatic diseases (n = 16)	Pancreatic cancer (n = 9)	P value
Bacteroides, %, median (range)	2.1 (0-26.1)	5.8 (0-46.4)	0.17
Bifidobacterium, %, median (range)	0 (0-0.5)	0.48 (0-4.5)	0.03
Clostridium cluster IV, %, mean ± SD	4.0 ± 3.3	3.6 ± 2.9	0.76
Clostridium cluster IX, %, mean ± SD	5.3 ± 3.6	6.4 ± 5.2	0.57
Clostridium cluster XI, %, median (range)	0 (0-0)	0 (0-0)
Clostridium cluster XVIII, %, median (range)	1.4 (0-3.9)	3.0 (0.8-14.9)	0.04
Clostridium subcluster XIVa, %, median (range)	3.8 (0-21.4)	6.0 (2.9-13.7)	0.32
Lactobacillales, %, mean ± SD	68.4 ± 19.3	59.7 ± 20.4	0.3
Prevotella, %, median (range)	4.2 (2.5-20.4)	4.0 (1.5-6.3)	0.3
Others, %, median (range)	2.0 (0-18.3)	0.3 (0-11.0)	0.3