Exosomal glypican-1 is elevated in pancreatic cancer precursors and can signal genetic predisposition in the absence of endoscopic ultrasound abnormalities

Table 1 Baseline demographic characteristics of the study population, n = 88

Cohort characteristics	MCL, n = 40	HR, n = 20	NLOD, n = 20	CG, n = 8	Total, n = 88	P value
Sex						NS
Male	14 (35.0)	9 (45.0)	9 (45.0)	2 (25.0)	34 (38.6)
Female	26 (65.0)	11 (55.0)	11 (55.0)	6 (75.0)	54 (61.4)
Age in yr	67.2 ± 8.9	48.7 ± 10.8	60.8 ± 7.4	53.8 ± 18.7	60.3 ± 12.6	< 0.001
BMI in kg/m2	24.9 ± 3.8	24.2 ± 3.9	29.9 ± 4.5	25.0 ± 4.0	25.9 ± 4.5	< 0.001
Smoking habits						NS
Never	27 (67.5)	13 (65.0)	12 (60.0)	7 (87.5)	59 (67.1)
Ex-smoker, > 5 yr	8 (20.0)	4 (20.0)	7 (35.0)	1 (12.5)	20 (22.7)
Active	5 (12.5)	3 (15.0)	1 (5.0)	0 (0.0)	9 (10.2)
Alcohol habits						NS
No	22 (55.0)	11 (55.0)	10 (50.0)	7 (87.5)	50 (56.8)
Yes	18 (45.0)	9 (45.0)	10 (50.0)	1 (12.5)	38 (43.2)
Family history of PDAC						< 0.001
No	34 (85.0)	7 (35.0)	18 (90.0)	8 (100.0)	67 (76.1)
Yes	6 (15.0)	13 (65.0)	2 (10.0)	0 (0.0)	21 (23.9)

Data are presented as n (%) or mean ± SD. BMI: Body mass index; CG: Control group; HR: Hereditary risk; MCL: Mucinous cystic lesion; NLOD: New-late onset diabetes mellitus; NS: Non-significant; PDAC: Pancreatic ductal adenocarcinoma; SD: Standard deviation.

Table 2 Characterization of hereditary risk group: Germline mutations and personal history of cancer, n = 20

	Lynch syndrome, n = 17 (85.0%)	Peutz-Jeghers syndrome, n = 1 (5.0%)	FPC, n = 2 (10.0%)
Germline mutation(s)
MLH1	7 (41.2)	-	-
MSH2	6 (35.3)	-	-
MLH1 + MSH2	1 (5.9)	-	-
MSH2 + MSH6	2 (11.7)	-	-
MSH6	1 (5.9)	-	-
STK11	-	1 (100.0)	-
Not identified	-		2 (100.0)
Personal history of cancer
No	11 (64.7)	1 (100.0)	2 (100.0)
Yes	6 (35.3)	0 (0.0)	0 (0.0)
Type of cancer
Colon and rectum	5 (83.3%)	-	-
Endometrium	1 (16.7)	-	-

FPC: Familial pancreatic cancer.

Table 3 Endoscopic ultrasound features of cystic lesions among the mucinous cystic lesions group, n = 40

Type of mucinous cyst, n (%)	MCN	1 (2.5)
	IPMN	39 (97.5)
Type of IPMN, n (%)	MD-IPMN	2 (5.1)
	BD-IPMN	35 (89.8)
	MT-IPMN	2 (5.1)
Number of cysts, n (%)	Single	16 (40.0)
	Multiple	24 (60.0)
Main cyst size in mm, mean ± SD		28.1 ± 14.4
Main cyst size in mm, median (IQR)		21.5 (16.3-40.5)
Septa, n (%)	Absent	10 (25.0)
	Present	30 (75.0)
Mural nodules, n (%)	Absent	36 (90.0)
	Present	4 (10.0)
Wall thickening, n (%)	Absent	37 (92.5)
	Present	3 (7.5)
MPD caliber, n (%)	Normal	34 (85.0)
	Dilated	6 (15.0)

BD-IPMN: Branch-duct IPMN; IPMN: Intraductal papillary mucinous neoplasm; IQR: Interquartile range; MCN: Mucinous cystic neoplasm; MD-IPMN: Main-duct IPMN; MPD: Main pancreatic duct; MT-IPMN: Mixed type.

Table 4 Clinical-pathological findings, treatment and follow-up of the study population, n = 88

	MCL, n = 40	HR, n = 20	NLOD, n = 20	CG, n = 8	Total, n = 88
PA in EUS
No	0 (0.0)	18 (90.0)	13 (65.0)	8 (100.0)	39 (44.3)
Yes	40 (100.0)	2 (10.0)	7 (35.0)	0 (0.0)	49 (55.7)
Type of PA
Cystic lesions	40 (100.0)	2 (100.0)	2 (28.6)	-	44 (89.8)
CP-like parenchymal changes	0 (0.0)	0 (0.0)	2 (28.6)	-	2 (4.1)
Solid lesions	0 (0.0)	0 (0.0)	0 (0.0)	-	0 (0.0)
Lipomatous transformation	0 (0.0)	0 (0.0)	3 (42.8)	-	3 (6.1)
PA with HRS / WF
No	20 (50.0)	2 (100.0)	7 (100.0)	-	29 (59.2)
Yes	20 (50.0)	0 (0.0)	0 (0.0)	-	20 (40.8)
EUS FNA/B
No	5 (12.5)	20 (100.0)	20 (100)	8 (100)	53 (60.2)
Yes	35 (87.5)	0 (0.0)	0 (0.0)	0 (0.0)	35 (39.8)
Inconclusive	13 (37.1)	-	-	-	13 (37.1)
Benign	19 (54.3)	-	-	-	19 (54.3)
Malignant	3 (8.6)1	-	-	-	3 (8.6)1
Surgery
No	25 (62.5)	20 (100)	20 (100)	8 (100)	73 (83.0)
Yes	15 (37.5)	0 (0.0)	0 (0.0)	0 (0.0)	15 (17.0)
Benign/Malignant	12 (80.0)/3 (20.0)	-/-	-/-	-/-	12 (80.0)/3 (20.0)
F-up in mo	33.3 ± 12.8	34.9 ± 7.1	27.7 ± 4.7	36.1 ± 5.1	32.6 ± 10.0
Cancer detection during F-up	1 (2.5)	0 (0.0)	0 (0.0)	0 (0.0)	1 (1.1)
Mortality	5 (12.5)	0 (0.0)	0 (0.0)	0 (0.0)	5 (5.7)

Data are presented as n (%) or mean ± SD.

¹One of 3 patients with malignant cytology was diagnosed during follow-up.

CG: Control group; CP: Chronic pancreatitis; EUS: Endoscopic ultrasound; FNA/B: Fine needle aspiration/biopsy; F-up: Follow up; HR: Hereditary risk; HRS: High risk stigmata; MCL: Mucinous cystic lesion; NLOD: New-late onset diabetes mellitus; NS: Non-significant; PA: Pancreatic abnormality; WF: Worrisome features.

Table 5 Biomarkers (glypican-1-positive circulating exosomes and carbohydrate antigen 19-9) profiles in the study groups, n = 88

Biomarker	MCL, n = 40	HR, n = 20	NLOD, n = 20	CG, n = 8	Total, n = 88	P value
GPC1+ crExos, %						< 0.001
Median	99.4	82.0	12.6	16.2	86.8
Min/Max	6.4/99.9	1.1/99.0	2.1/93.9	1.2/24.5	1.1/99.9
IQR	94.9-99.8	28.9-98.2	5.2-63.4	6.6-20.1	18.6-99.4
CA 19-9 in U/mL						NS
Median	20.4	16.9	18.8	30.6	18.4
Min/Max	8.5/206.6	8.3/28.0	12.8-48.7	13.7-69.6	8.3/206.6
IQR	14.0-35.8	11.7-18.1	16.6-23.4	15.0-40.8	14.2-27.7

P value for Kruskal-Wallis test. CA 19-9: Carbohydrate antigen 19-9; CG: Control group; HR: Hereditary risk; IQR: Interquartile range; GPC1⁺ crExos: Glypican-1 positive circulating exosomes; Max: Maximum; MCL: Mucinous cystic lesion; Min: Minimum; NLOD: New-late onset diabetes mellitus; NS: Non-significant.

Table 6 Size and concentration of exosomes according to the study groups, n = 88

	MCL, n = 40	HR, n = 20	NLOD, n = 20	CG, n = 8	Total, n = 88	P value
Size in nm						0.012
Median	82.5	94.7	100.7	72.8	90.5
IQR	72.8-97.6	83.0-109.7	89.2-110.1	67.0-102.8	77.9-103.2
Concentration as particles E + 10/mL						< 0.001
Median	3.48	3.36	6.05	3.30	4.10
IQR	1.88-5.08	2.51-4.37	4.83-7.22	2.91-4.20	2.60-5.69

P value for Kruskal-Wallis test. CG: Control group; HR: Hereditary risk; IQR: Interquartile range; MCL: Mucinous cystic lesion; NLOD: New-late onset diabetes mellitus.