Identification of novel autoantibodies based on the protein chip encoded by cancer-driving genes in detection of esophageal squamous cell carcinoma

Figures & data

Figure 1. The design of this study. ESCC: esophageal squamous cell carcinoma; NC: normal control; ELISA: enzyme-linked immunosorbent assay.

Table 1. Clinical characteristics of ESCC patients and controls in verification and validation phase.

	Training cohort (n = 260)		Validation cohort (n = 250)
Variables	ESCC	NC	ESCC	NC	P
Age(year)					0.055
Range	41–87	41–88	41–80	40–79
Median (Q1, Q3)	65.5 (58.5, 70.3)	66.0 (59.8, 71.0)	63.0 (59.9, 68.0)	63.0 (59.0, 68.0)
Sex, n (%)					0.037*
Male	95(73.1)	95(73.1)	76(60.8)	76(60.8)
Female	35(26.9)	35(26.9)	49(39.2)	49(39.2)
Histological grade					0.065
High(grade 1)	9(6.9)		15(12.0)
Middle(grade 2)	44(33.8)		46(36.8)
Low(grade 3)	20(15.4)		50(40.0)
Site of tumor					0.334
Upper esophagus	16(12.3)		22(17.6)
Middle esophagus	70(53.8)		65(52.0)
Lower esophagus	41(31.5)		31(24.8)
TNM stage, n (%)					0.001*
Stage 0	7(5.4)		7(5.6)
Stage I	22(16.9)		47(37.6)
Stage II	23(17.7)		38(30.4)
Stage III	35(26.9)		24(19.2)
Stage IV	12(9.2)		5(4.0)
Depth of tumor invasion					0.054
T0	5(3.8)		7(5.6)
T1	29(22.3)		43(34.4)
T2	19(14.6)		20(16.0)
T3	37(28.5)		48(38.4)
T4	12(9.2)		0(0.00)
Lymph node metastasis					0.000*
Negative	47(36.2)		86(68.8)
Positive	66(50.7)		32(25.6)
Distant metastasis					0.052
No	90(69.2)		116(92.8)
Yes	11(8.5)		5(4.0)

ESCC: esophageal squamous cell carcinoma; NC: normal control; Q1: smaller quartile, Q3: larger quartile, P means comparison between ESCC patients in training cohorts and validation cohorts, P* < 0.05

Table 2. Diagnostic value of 12 anti-TAAs autoantibodies in training and validation cohort for ESCC detection.

	Training cohort					Validation cohort
TAAbs	Se (%)	Sp (%)	AUC	95%CI	Accuracy (%)	Se (%)	Sp (%)	AUC	95%CI	Accuracy (%)
GNA11	59.2	96.9	0.84	0.80–0.89	75.8	81.6	78.4	0.82	0.77–0.88	78.8
PTEN	60.0	66.9	0.65	0.59–0.72	61.1	68.0	56.0	0.64	0.57–0.70	61.2
P53	83.1	64.6	0.76	0.70–0.82	71.5	63.2	74.4	0.75	0.68–0.81	67.6
SRSF2	67.7	52.3	0.61	0.54–0.68	58.5	78.4	45.6	0.64	0.57–0.71	60.0
GNAS	87.7	26.9	0.58	0.51–0.65	54.6	69.6	52.0	0.60	0.53–0.67	58.8
ACVR1B	88.5	36.2	0.59	0.52–0.66	54.6	96.8	28.8	0.61	0.54–0.68	56.8
CASP8	40.8	74.6	0.58	0.51–0.65	55.0	45.6	72.8	0.58	0.51–0.65	55.6
DAXX	40.8	88.5	0.63	0.56–0.70	59.2	36.8	92.8	0.59	0.52–0.66	57.6
PDGFRA	45.4	83.1	0.66	0.59–0.73	62.7	76.0	45.6	0.64	0.57–0.71	60.0
FBXW7	24.6	86.9	0.50	0.43–0.57	49.2	39.2	79.2	0.55	0.48–0.62	53.6
MEN1	33.8	85.4	0.67	0.50–0.64	53.9	54.0	64.0	0.58	0.51–0.65	58.8
EGFR	28.5	90.8	0.53	0.46–0.60	50.8	56.0	59.2	0.56	0.49–0.63	57.2

Se: sensitivity, Sp: specificity, AUC: area under the receiver operating characteristic curve; CI: confidence interval.

Figure 2. The expression levels of 12 anti-TAAs autoantibodies in ESCC patients and healthy controls. (a) Scatter plots of the expression levels of autoantibodies in training cohort, (b) scatter plots of the expression levels of autoantibodies in validation cohort. Lines represented median and quartile range; E, ESCC patients; N normal controls.

Figure 3. Diagnostic performance of 12 anti-TAAs in training cohort for ESCC detection.

Figure 4. Diagnostic performance of 12 anti-TAAs in validation cohort for ESCC detection.

Table 3. Diagnostic value of the anti-TAAs autoantibodies panel for ESCC patients with different stages.

Group	n	Se (%)	Sp (%)	Accuracy (%)	AUC	95%CI	p^a	p^b	PPV (%)	NPV (%)	+LR	-LR
Training cohort
All stage	130	71.5	93.8	79.6	0.88	0.84–0.92			82.9	76.9	11.6	0.3
TNM0-II	52	73.1	93.9	81.3	0.90	0.85–0.95	0.540	0.066	65.5	88.7	11.9	0.3
TNMIII-IV	81	63.8	89.2	73.4	0.81	0.73–0.89	0.137		60.0	86.6	5.9	0.4
T0-T2	53	77.4	80.8	79.2	0.87	0.81–0.91	0.786	0.835	63.6	85.9	4.0	0.3
T3-T4	49	73.5	90.8	83.0	0.86	0.80–0.91	0.641		87.5	89.4	8.0	0.3
Lymph node(-)	47	74.5	93.8	81.9	0.92	0.87–0.96	0.191	0.080	63.5	92.4	12.1	0.3
Lymph node(+)	66	63.6	97.7	79.7	0.85	0.78–0.91	0.447		58.0	85.5	27.6	0.4
Distant metastasis(-)	90	67.8	93.9	79.1	0.86	0.81–0.91	0.558		76.2	63.6	11.1	0.3
Distant metastasis(+)	11	90.9	73.8	82.3	0.87	0.81–0.92	0.859	0.865	25.9	96.5	3.5	0.3
Validation cohort
All stage	125	77.6	81.6	70.8	0.85	0.80–0.90			87.1	64.4	4.2	0.3
TNM0-II	92	81.5	81.6	76.5	0.87	0.83–0.92	0.546	0.071	84.7	73.4	4.4	0.2
TNMIII-IV	29	65.5	82.4	57.6	0.77	0.67–0.86	0.150		47.4	85.3	3.7	0.4
T0-T2	70	84.3	81.6	78.5	0.88	0.82–0.92	0.403	0.184	80.4	77.9	4.6	0.2
T3-T4	48	77.1	75.2	79.8	0.82	0.76–0.88	0.498		71.0	81.7	3.1	0.3
Lymph node(-)	86	87.2	76.0	76.8	0.87	0.82–0.92	0.571	0.237	83.6	74.4	3.6	0.2
Lymph node(+)	32	68.7	83.2	82.2	0.81	0.72–0.89	0.440		59.1	85.9	4.1	0.4
Distant metastasis(-)	116	77.6	81.6	86.7	0.85	0.80–0.90	0.863		91.2	62.7	4.2	0.3
Distant metastasis(+)	5	80.0	82.4	67.6	0.75	0.67–0.83	0.460	0.460	10.0	96.7	4.6	0.2

P^ameans comparison between TNM stage or T stage or lymph node or Distant metastasis and all stage with the method of De Long et al. (1988)

P^b means comparison between TNM0-II and TNMIII-IV stage or T0-T2 and T3-T4 or lymph node (-) and lymph node (+) or Distant metastasis (-) and Distant metastasis (+). Se: sensitivity, Sp: specificity, AUC: area under the receiver operating characteristic curve; CI: confidence interval; PPV: positive predictive value; NPV: negative predictive value; +LR: positive likelihood ration; -LR: negative likelihood ration.

Figure 5. The value of the model for detecting ESCC with different stages in two cohorts. (a) The prediction model for ESCC detection in training cohort. (b) The prediction model for ESCC detection in validation cohort.