Serum Glial Cell Line-derived Neurotrophic Factor (sGDNF) is a Novel Biomarker in Predicting Cirrhosis in Patients With Chronic Hepatitis B

doi:10.21203/rs.3.rs-740620/v1

We assessed the potential of glial cell line-derived neurotrophic factor (GDNF) as a useful biomarker to predict cirrhosis in chronic hepatitis B (CHB) patients.A total of 735 patients from two medical centers (385 CHB patients and 350 healthy controls) were included to determine the association of serum and tissue GDNF levels with biopsy-proven cirrhosis. The diagnostic accuracy of serum GDNF (sGDNF) was estimated and compared with other indices of cirrhosis. We showed significantly higher levels of sGDNF in CHB patients with fibrosis (28.4 pg/ml vs. 11.6 pg/ml in patients without) and patients with cirrhosis (33.8 pg/ml vs. 23.5 pg/ml in patients without). The areas under receiver operating curve (AUROCs) of sGDNF were 0.83 (95% confidence interval (CI): 0.80-0.87) for predicting liver fibrosis and 0.84 (95% CI: 0.79-0.89) for cirrhosis. Findings from serum protein level and hepatic mRNA expression were consistent. Using the best cutoff to predict cirrhosis, we categorized the patients into sGDNF-high and sGDNF-low groups. The sGDNF-high group had significantly larger Masson’s trichrome and reticulin staining-positive area, higher Scheuer score and METAVIR fibrosis stage (all p<0.001) but not steatosis. On multivariable regression, sGDNF was independently associated with cirrhosis with an odds ratio at 6.98 (95% CI: 1.10-17.94). Finally, we demonstrated that sGDNF outperformed AST to platelet ratio index, FIB-4, fibroscore, forn index, and fibrometer in differentiating F4 vs. F3.

Using serum, tissue mRNA, and biopsy data, our study revealed a significant potential of sGDNF as a novel non-invasive biomarker for cirrhosis in CHB patients.

Clinical Pharmacology

glial cell line-derived neurotrophic factor (GDNF)

cirrhosis

diagnostic accuracy

non-invasive biomarker

This preprint is available for

download as a PDF

.

Table 1A. Clinical demographics and clinical characterics of patients in non-fibrosis and fibrosis groups

	Total (N=694)	Non-fibrosis (N=376)^*	Fibrosis(N=318)^**	p value
Age, yr	43 (34, 55)	45 (39, 62)	40 (32, 48)	0.000
Male, N (%)	360 (52)	221 (61)	113 (34)	0.000
ALT, U/L	21 (11, 53)	42 (28, 168)	71 (30, 195)	0.000
AST, U/L	26 (19, 46)	28 (23, 132)	46 (28, 103)	0.000
BUN, mmol/L	4.4 ± 1.3	3.9 ± 1.0	4.7 ± 1.3	0.001
Cr, μmol/L	67 (57, 79)	63 (56, 77)	70 (58, 80)	0.001
RBC, 10¹²/L	4.7 ± 0.5	4.7 ± 0.5	4.8 ± 0.5	0.108
PLT, 10⁹/L	199 ± 61	227 ± 55	183 ± 59	0.000
WBC, 10⁹/L	5.9 ± 3.1	6.4 ± 4.6	5.5 ± 1.5	0.105
Glucose, mmol/L	4.8 (4.4, 5.2)	5.1 (4.7, 5.1)	4.9 (4.5, 5.5)	0.017
sGDNF, pg/ml	25.2 (11.4, 29.5)	11.6 (7.2, 22.1)	28.4 (26.2, 31.6)	0.000

non-fibrosis is defined by health controls and F0;fibrosis is defined by F1-F4

Table 1B. Clinical demographics and clinical characterics of patients in non-cirrhosis and cirrhosis groups

	Total (N=694)	Non-cirrhosis (N=661)^#	cirrhosis(N=33)^##	p value
Age, yr	42 (34, 52)	42 (33, 52)	40 (35, 49)	0.605
Male, N (%)	360 (52)	335 (51)	25 (76)	0.000
ALT, U/L	21 (11, 53)	16 (10, 51)	107 (53, 448)	0.000
AST, U/L	26 (19, 46)	23 (19, 40)	99 (47, 376)	0.000
BUN, mmol/L	4.7 ± 1.5	4.7 ± 1.5	4.7 ± 1.3	0.858
Cr, μmol/L	67 (57, 79)	79 (67, 88)	67 (60, 80)	0.374
RBC, 10¹²/L	4.7 ± 0.5	4.7 ± 0.5	4.5 ± 0.5	0.021
PLT, 10⁹/L	199 ± 61	203 ± 60	141 ± 42	0.000
WBC, 10⁹/L	5.9 ± 3.0	5.9 ± 3.1	4.9 ± 1.5	0.001
Glucose, mmol/L	4.8 (4.4, 5.2)	4.7 (4.4, 5.2)	4.9 (4.4, 5.5)	0.462
sGDNF, pg/ml	25.2 (11.4, 29.5)	23.5 (10.9, 29.5)	33.8 (29.3, 39.4)	0.000

non-cirrhosis is defined by health controls and F0-F3; cirrhosis is defined by F4

The data are expressed as mean ± standard deviation or median (25%-75%).

ALT, alanine aminotransferase; AST, aspartate aminotransferase; BUN, blood urea nitrogen; Cr, creatinine; PLT, platelet count; RBC, red blood cell; sGDNF, serum GDNF; WBC, white blood cell.

Table 2. Characteristics of study participants relative to their sGDNF levels.

	Total (N=344)	sGDNF-low (N=192)	sGDNF-high (N=152)	p value
Steatosis, N (%)
Negative	209 (60.8)	114 (59.4)	95 (62.5)	0.556
Positive	135 (39.2)	78 (40.6)	57 (37.5)
G stage, N (%)
0-1	32 (9.3)	20 (10.4)	12 (7.9)	0.057
2	176 (51.2)	105 (54.7)	71 (46.7)
3-4	136 (39.5)	67 (34.9)	69 (45.4)
F stage, N (%)
0	26 (7.6)	17 (8.9)	9 (5.9)	0.000
1	84 (24.4)	55 (26.8)	29 (19.1)
2	133 (38.7)	80 (41.7)	53 (34.9)
3	68 (19.8)	35 (18.2)	33 (21.7)
4	33 (9.6)	5 (2.6)	28 (18.4)
Pathology staining, (%)
Masson’s trichrome staining (N=157)	5.0 (2.2, 9.0)	3.1 (1.2, 7.0)	6.3 (3.1, 11.4)	0.001
Reticulin staining (N=224)	2.3 (0.9, 4.5)	1.8 (0.7, 4.3)	2.9 (1.5, 5.0)	0.023

The data are expressed as median (25-75%). p for trends determined through the linear-by linear association test. Liver histopathology of patients with G0, G1, G2, G3, and G4 according to the Scheuer scoring system. G1, portal inflammation; G2, mild piecemeal necrosis; G3, moderate piecemeal necrosis; G4, severe piecemeal necrosis and bridging necrosis. METAVIR scoring system. F0, no fibrosis; F1, Portal fibrosis without septa; F2, Septal fibrosis (portal-portal); F3, Septal fibrosis (portal-central); F4, Cirrhosis.

Table 3. Univariate and multivariate analyses producing odds ratio for significant F4 stage in chronic HBV patients.

Variables	Univariate analysis RR (95% C.I)	p value	Multivariate analysis RR (95% C.I)	p value
AFP	1.02 (1.01-1.04)	0.001
Alb	0.84 (0.77-0.92)	0.000	*0.85 (0.74-0.97)*	*0.019*
ALP	1.01 (1.00-1.01)	0.034
ALT	1.00 (1.00-1.00)	0.016
AST	1.00 (1.00-1.00)	0.010
CHE	1.00 (1.00-1.00)	0.001
CⅣ	1.01 (1.00-1.01)	0.001
G	14.83 (4.51-48.76)	0.000	*5.55 (1.56-19.67)*	*0.008*
Hb	0.96 (0.93-0.99)	0.016
Neutrophil	0.60 (0.40-0.88)	0.010
PLT	0.98 (0.98-0.99)	0.001
PTA	0.98 (0.96-0.99)	0.009
RBC	0.16 (0.06-0.41)	0.000
sGDNF	13.33 (4.76-37.34)	0.000	*6.98 (1.10-17.94)*	*0.036*
TBA	1.01 (1.00-1.02)	0.009
TBil	1.04 (1.02-1.07)	0.001
WBC	0.60 (0.43-0.86)	0.005

All baseline covariates were included in the univariable analysis (two-sided p value < 0.05). Only covariates significantly associated with F4 in the univariable analysis (two-sided p value < 0.05) are shown and included in the multivariable model. Alb, albumin, AFP, alpha fetoprotein; ALP, alkaline phosphatase; ALT, alanine amiotransferase; AST, aspartate aminotransferase; CHE, cholinesterase; CIV, type IV collagen; Hb, hemoglobin; PLT, platelet count; PTA, prothrombin activity; RBC, red blood cell count. TBA, total bile acid; TBil, total bilirubin; WBC, white blood cell count.

Table 4. Areas under the receiver operating characteristic curve for the diagnostic accuracy of sGDNF, APRI, FIB-4, Fibrotest, Forn index, Hepascore, and Fibrometer for the diagnosis of histologic fibrosis stage F4.

	AUROC (95% CI)	Cut-off	Sensitivity (%)	Specificity (%)	PPV (%)	NPV (%)	Youden	vs sGDNF p value
Primary Analysis F0-3 vs F4
sGDNF	0.78 (0.73-0.82)	28.74	84.85	60.65	18.5	97.4	0.4550
APRI	0.68 (0.63-0.73)	0.41	93.94	38.89	14.2	98.3	0.3283	0.1417
FIB-4	0.71 (0.66-0.76)	1.12	90.91	44.05	14.7	97.9	0.3496	0.3216
fibrotest	0.70 (0.65-0.75)	0.89	63.64	70.10	18.4	94.8	0.3373	0.1998
forn index	0.73 (0.68-0.78)	3.94	93.94	40.19	14.3	98.4	0.3413	0.4837
hepascore	0.67 (0.62-0.72)	0.80	75.76	59.16	16.4	95.8	0.3492	0.1234
fibrometer	0.69 (0.64-0.74)	2.54	75.76	56.91	15.7	95.7	0.3267	0.1856
Second analysis F3 vs F4
sGDNF	0.74 (0.65-0.83)	33.43	63.64	83.58	65.6	82.4	0.4722
APRI	0.55 (0.44-0.65)	0.41	93.94	25.76	38.7	89.5	0.1970	0.0124
FIB-4	0.55 (0.44-0.65)	2.20	57.57	61.19	42.2	74.5	0.1877	0.0181
fibrotest	0.61 (0.50-0.70)	0.89	63.64	59.70	43.8	76.9	0.2334	0.0802
forn index	0.57 (0.47-0.67)	5.45	63.64	50.75	38.9	73.9	0.1438	0.0368
hepascore	0.55 (0.45-0.65)	0.80	75.76	40.30	38.5	77.1	0.1606	0.0211
fibrometer	0.53 (0.43-0.64)	1.69	87.88	25.37	36.7	81.0	0.1325	0.0133

AUROC of sGDNF vs APRI, FIB-4, Fibrotest, Forn index, Hepascore, Fibrometer via DeLong test.

NPV, negative predictive value; PPV, positive predictive value; Se, sensitivity; Sp, specificity.

APRI, aspartate transaminase-platelet ratio index;

FIB-4, fibrosis-4 index.

APRI: (AST(U/L)/upper normal limit)×100/platelets (10⁹/L) .

FIB-4: age (years) × AST (U/L) / (PLT (10⁹/L)) × (ALT(U/L)^1/2) .

Fibrometer: -0.007 PLT (G/L)-0.049 PI (%) + 0.012AST (U/L) + 0.005 α2M (mg/dL) + 0.021HA (μg/L) - 0.270 urea (mmol/L) + 0.027 age (yr) + 3.718 .

Forn index: 7.811-3.131 × ln(PLT(10⁹/L)) + 0.781 × ln(GGT(U/L)) + 3.467 × ln(age)-0.014 × (cholesterol (mg/dl) .

Hepascore:

y = exp[4.185818 - (0.0249 × age) + (0.7464 × sex) + (1.0039 × α2M g/L) + (0.0302×HA μg/L) + (0.0691× bilirubin μmol/L) - (0.0012×GGT U/L)], male =1, female=0 .

Fibrotest: 4.467×log (α2M (mg/dL)) -1.357 × log Hap (g/L) + 1.017 log GGT (U/L) + 0.0281× age+1.737× log TBil (μmol/L) - 1.184 × apoA-I (g/L) + 0.301× (sex)-5.540, male =1, female=0 .

Serum Glial Cell Line-derived Neurotrophic Factor (sGDNF) is a Novel Biomarker in Predicting Cirrhosis in Patients With Chronic Hepatitis B

Status:

Version 1

Abstract

Figures

Full Text

Tables

Status:

Version 1