Comprehensive Profiling of Intrahepatic Cholangiocarcinoma

This study included five ICC patient cohorts. (1) The FU-iCCA cohort enrolled 262 ICC patients from Zhongshan Hospital, Fudan University [22 (link)]. Multi-omics data of this cohort, including data of whole-exome sequencing (WES), RNA sequencing, and proteome, were analyzed. (2) The second cohort recruited 259 patients with pathologically confirmed ICC undergoing curative resection in Zhongshan Hospital (ZSH cohort) from June 2012 to December 2017. All enrolled patients received no anti-cancer therapy prior to surgery. All tumor specimens from the ZSH cohort were formalin-fixed and paraffin-embedded and collected for tissue microarrays (TMA) construction. The ZSH cohort was used to validate the findings from the FU-iCCA cohort. The baseline characteristics of the FU-iCCA cohort and the ZSH cohort are detailed in Table 1. Serological tests were performed within 3 days before the operation. The clinical stage was evaluated based on the American Joint Committee on Cancer (AJCC) 8th edition [23 (link)]. (3) The third cohort included five ICC patients from the single-cell RNA sequencing dataset GSE138709 [24 (link)]. (4) We extracted the single-cell data of ten ICC patients for the immune checkpoint blockade (ICB) clinical trial (ICB cohort) from GSE151530 and divided them into two groups (baseline group and ICB-treated group) [25 (link)]. (5) The fifth cohort recruited an independent cohort of 33 ICC patients receiving surgical resection from January 2019 to June 2019 in Zhongshan Hospital. CD73 expression between matched tumor and para-tumor tissues was compared by RT-PCR assays.Table 1

Correlation between CD73 expression and clinical features of patients enrolled

Characteristics	FU-iCCA cohort (n = 244)					ZSH cohort (n = 259)
	Patients		CD73 expression			Patients		CD73 expression
	No	%	Low	High	P value	No	%	Low	High	P value
All patients	244	100	140	104		259	100	146	113
Sex
Female	106	43.4	62	44	0.758	99	38.2	67	32	0.004
Male	138	56.6	78	60		160	61.8	79	81
Age
≤ 60	113	46.3	67	46	0.574	125	48.3	69	56	0.714
> 60	131	53.7	73	58		134	51.7	77	57
HBsAg
Negative	179	73.4	104	75	0.705	179	69.1	97	82	0.290
Positive	65	26.6	36	29		80	30.9	49	31
Liver cirrhosis
No	222	91.0	128	94	0.778	186	71.8	104	82	0.813
Yes	22	9.0	12	10		73	28.2	42	31
Vascular invasion
No	141	57.8	87	54	0.110	185	71.4	115	70	0.003
Yes	103	42.2	53	50		74	28.6	31	43
LN metastasis
No	194	79.5	114	80	0.389	207	79.9	124	83	0.022
Yes	50	20.5	26	24		52	20.1	22	30
Tumor size
≤ 5 cm	108	44.3	63	45	0.788	116	44.8	71	45	0.158
> 5 cm	136	55.7	77	59		143	55.2	75	68
CA199
≤ 37 U/mL	109	44.7	75	34	0.001	116	44.8	76	40	0.008
> 37 U/mL	135	55.3	65	70		143	55.2	70	73
CEA
≤ 5 ng/mL	185	75.8	116	69	0.003	189	73.0	118	71	0.001
> 5 ng/mL	59	24.2	24	35		70	27.0	28	42
AJCC 8th
I–II	154	63.1	96	58	0.040	203	78.4	122	81	0.021
III–IV	90	36.9	44	46		56	21.6	24	32

Data in bold indicated statistical significance

ZSH cohort Zhongshan Hospital cohort, LN Lymph node, CEA Carcinoembryonic antigen, AJCC American Joint Committee on Cancer