Alpha-fetoprotein Negative Hepatocellular Carcinoma Research Articles

Prediction of survival and analysis of prognostic factors for patients with AFP negative hepatocellular carcinoma: a population-based study

PurposeHepatocellular carcinoma (HCC) has a poor prognosis, and alpha-fetoprotein (AFP) is widely used to evaluate HCC. However, the proportion of AFP-negative individuals cannot be disregarded. This study aimed to establish a nomogram of risk factors affecting the prognosis of patients with AFP-negative HCC and to evaluate its diagnostic efficiency.Patients and methodsData from patients with AFP-negative initial diagnosis of HCC (ANHC) between 2004 and 2015 were collected from the Surveillance, Epidemiology, and End Results database for model establishment and validation. We randomly divided overall cohort into the training or validation cohort (7:3). Univariate and multivariate Cox regression analysis were used to identify the risk factors. We constructed nomograms with overall survival (OS) and cancer-specific survival (CSS) as clinical endpoint events and constructed survival analysis by using Kaplan-Meier curve. Also, we conducted internal validation with Receiver Operating Characteristic (ROC) analysis and Decision curve analysis (DCA) to validate the clinical value of the model.ResultsThis study included 1811 patients (1409 men; 64.7% were Caucasian; the average age was 64 years; 60.7% were married). In the multivariate analysis, the independent risk factors affecting prognosis were age, ethnicity, year of diagnosis, tumor size, tumor grade, surgery, chemotherapy, and radiotherapy. The nomogram-based model related C-indexes were 0.762 (95% confidence interval (CI): 0.752–0.772) and 0.752 (95% CI: 0.740–0.769) for predicting OS, and 0.785 (95% CI: 0.774–0.795) and 0.779 (95% CI: 0.762–0.795) for predicting CSS. The nomogram model showed that the predicted death was consistent with the actual value. The ROC analysis and DCA showed that the nomogram had good clinical value compared with TNM staging.ConclusionThe age(HR:1.012, 95% CI: 1.006–1.018, P-value < 0.001), ethnicity(African-American: HR:0.946, 95% CI: 0.783–1.212, P-value: 0.66; Others: HR:0.737, 95% CI: 0.613–0.887, P-value: 0.001), tumor diameter(HR:1.006, 95% CI: 1.004–1.008, P-value < 0.001), year of diagnosis (HR:0.852, 95% CI: 0.729–0.997, P-value: 0.046), tumor grade(Grade 2: HR:1.124, 95% CI: 0.953–1.326, P-value: 0.164; Grade 3: HR:1.984, 95% CI: 1.574–2.501, P-value < 0.001; Grade 4: HR:2.119, 95% CI: 1.115–4.027, P-value: 0.022), surgery(Liver Resection: HR:0.193, 95% CI: 0.160–0.234, P-value < 0.001; Liver Transplant: HR:0.102, 95% CI: 0.072–0.145, P-value < 0.001), chemotherapy(HR:0.561, 95% CI: 0.471–0.668, P-value < 0.001), and radiotherapy(HR:0.641, 95% CI: 0.463–0.887, P-value:0.007) were independent prognostic factors for patients with ANHC. We developed a nomogram model for predicting the OS and CSS of patients with ANHC, with a good predictive performance.

Construction and Validation of a Novel Nomogram Predicting Recurrence in Alpha-Fetoprotein-Negative Hepatocellular Carcinoma Post-Surgery Using an Innovative Liver Function-Nutrition-Inflammation-Immune (LFNII) Score: A Bicentric Investigation.

We developed a nomogram based on the liver function, nutrition, inflammation, and immunity (LFNII) score to predict recurrence-free survival (RFS) post-resection in patients with hepatocellular carcinoma (HCC) exhibiting alpha-fetoprotein (AFP) negativity (AFP ≤20 ng/mL). Clinical data of 661 patients diagnosed with alpha-fetoprotein-negative hepatocellular carcinoma (AFP-NHCC) who underwent surgical resection at two medical centers between 2012 and 2021 were collected. A total of 462 and 199 patients served as the training and validation sets, respectively. Pre-operative blood markers were collected and analyzed for LFNII. The LFNII score was formulated using the least absolute shrinkage and selection operator Cox regression model. A nomogram model was developed using the training set to incorporate other relevant clinicopathological indicators and predict postoperative recurrence. Model discrimination was assessed using the receiver operating characteristic curve, calibration was evaluated using a calibration curve, and clinical applicability was assessed using clinical decision curve analysis. A comparison with liver cancer staging was performed using the nomogram model. Finally, a cohort study was conducted to validate our findings. We derived the LFNII scores from nine indicators. Elevated LFNII scores correlated with unfavorable clinicopathological features. The LFNII score area under the curve revealed superior predictive efficacy at 1-, 2-, and 5-year RFS intervals, with values of 0.675, 0.658, and 0.633, respectively. Multivariate Cox analysis revealed that a high LFNII score independently increased RFS risk in patients with AFP-NHCC. The C-index of the LFNII-nomogram model was 0.686 (95% confidence interval [CI], 0.651-0.721). The nomogram model's clinical application value surpassed that of standard HCC staging systems. The LFNII score-derived nomogram effectively predicted the RFS of patients with AFP-NHCC after curative resection.

[Analysis of the diagnostic efficiency of combining multiple laboratory hematological indicators in alpha-fetoprotein-negative hepatocellular carcinoma].

Objective: To establish a diagnostic model for alpha-fetoprotein-negative hepatocellular carcinoma (AFP-NHCC) by combining multiple laboratory hematological indicators and explore its clinical diagnostic efficiency. Methods: A total of 124 inpatients, including 110 males and 14 females, aged 57 (51, 66) years, who were first diagnosed with AFP-NHCC in the PLA General Hospital were included from December 2011 to June 2017. Meanwhile, 331 cases of non-HCC were enrolled as the control group, including 279 males and 52 females, aged 58 (51, 63) years old, with 47 cases of hepatitis B virus (HBV) infection, 40 cases of liver cirrhosis, 64 cases of hepatic hemangioma or cysts, 7 cases of liver nodules, 8 cases of fatty liver, 146 cases of non-liver disease and 19 health controls. Subjects in the AFP-NHCC group and the control group were divided into a training group and a validation group. A total of 196 subjects were involved in the training group, including 103 AFP-NHCC patients and 93 non-HCC patients (19 healthy controls, 25 patients with HBV infection, 22 patients with liver cirrhosis, 23 patients with hepatic hemangioma or cyst, and 4 patients with liver nodules). The differences in laboratory parameters were analyzed, and a diagnostic model of AFP-NHCC under different AFP levels was established. Likewise, 259 subjects, including 113 patients with liver disease, were involved in the validation group to verify the diagnostic efficiency of the model for AFP-NHCC. The receiver operating characteristic (ROC) curve was used to analyze the sensitivity and specificity of different models, and the area under the curve (AUC) was calculated to evaluate the diagnostic performance of different models. Results: In the training group, the indicators of AFP-NHCC diagnostic model included platelet (PLT), prothrombin activity (PTA), serum albumin (ALB), prothrombin time (PT) and carbohydrate antigen 19-9 (CA19-9), and the AUC of the model was 0.848 (95%CI: 0.786-0.911) when AFP≤5 μg/L. Similarly, the indicators of AFP-NHCC diagnostic model included PLT, PTA, ALB, PT and hematocrit (HCT), and the AUC of the model was 0.839 (95%CI: 0.780-0.897) when AFP≤10 μg/L. When AFP≤20 μg/L, the indicators of AFP-NHCC diagnostic model contained PLT, PTA, ALB, PT, HCT and AFP, and the AUC of the model was 0.866 (95%CI: 0.815-0.917). The AUC values of these three models were higher than those of AFP and CA19-9 alone for the diagnosis of AFP-NHCC [0.634 (95%CI: 0.560-0.709), 0.691 (95%CI:0.620-0.761), all P<0.05]. The indicators screened by these three models were combined to establish the final diagnostic model, and the AUC of the model was 0.873 (95%CI: 0.824-0.923), with the sensitivity of 78.6% (81/103) and the specificity of 81.7% (76/93). In the validation group, the predictive AUC of the final model in liver disease patients was 0.892 (95%CI: 0.832-0.951), with the sensitivity of 100% (21/21) and the specificity of 71.7% (66/92), while in the total validation population, the predictive AUC was 0.931 (95%CI: 0.890-0.972), with the sensitivity of 100.0% (21/21) and the specificity of 75.6% (180/238). Conclusion: The final diagnostic model includes PLT, PTA, ALB, PT, HCT, CA19-9 and AFP, which has higher sensitivity and specificity, and has good diagnostic efficiency for the clinical diagnosis of AFP-NHCC.

Comparison of IDEAL-IQ and IVIM-DWI for Differentiating between Alpha Fetoprotein-Negative Hepatocellular Carcinoma and Focal Nodular Hyperplasia

Background: To compare the differential diagnostic value of iterative decomposition of water and fat with the echo asymmetrical and least-squares estimation quantitation sequence (IDEAL-IQ) with that of intravoxel incoherent motion diffusion-weighted imaging (IVIM DWI) in differentiating between alpha fetoprotein (AFP)-negative hepatocellular carcinoma (HCC) and focal nodular hyperplasia (FNH). Materials and Methods: A total of 28 AFP-negative HCC cases and 15 FNH cases were scanned using the IDEAL-IQ and IVIM-DWI magnetic resonance imaging (MRI) protocols. Two radiologists independently assessed the fat fraction (FF) and the iron level surrogate (R2*) derived from the IDEAL-IQ images and the apparent diffusion coefficient (ADC), pure diffusion coefficient (D), pseudo-diffusion coefficient (D*), and perfusion fraction (f) derived from the IVIM-DWI images. The intraclass correlation coefficients (ICC) were estimated to evaluate the agreement repeatability between the two readers. The area under the curve (AUC) of a receiver operating characteristics curve was used to compare the diagnostic efficiency of the parameters extracted from the two techniques. Results: The lesions in the HCC group had significantly higher FF than the FNH group (8.284 ± 5.756 vs. 2.559 ± 1.247, P < 0.05). ADC and D values were lower in the HCC lesions when compared with the FNH lesions (1.310 ± 0.253 and 0.909 ± 0.192, respectively, vs. 1.624 ± 0.304 and 1.230 ± 0.314; ×10−3 mm2 /s). The FF parameter had the highest AUC (0.923) followed by D (0.864) and ADC (0.854). Conclusions: FF derived from IDEAL-IQ, and ADC and D derived from IVIM-DWI were able to differentiate AFP-negative HCC from FNH. IDEAL-IQ showed better performance for the differentiation of FNH from HCC than the IVIM-DWI-derived parameters.

N‐glycan fingerprint predicts alpha‐fetoprotein negative hepatocellular carcinoma: A large‐scale multicenter study

Alpha-fetoprotein (AFP)-negative hepatocellular carcinoma (ANHCC) patients account for more than 30% of the whole entity of HCC patients and are easily misdiagnosed. This three-phase study was designed to find and validate new ANHCC N-glycan markers which identified from The Cancer Genome Atlas (TCGA) database and noninvasive detection. Differentially expressed genes (DEGs) of N-glycan biosynthesis and degradation related genes were screened from TCGA database. Serum N-glycan structure abundances were analyzed using N-glycan fingerprint (NGFP) technology. Totally 1340 participants including ANHCC, chronic liver diseases and healthy controls were enrolled after propensity score matching (PSM). The Lasso algorithm was used to select the most significant N-glycan structures abundances. Three machine learning models [random forest (RF), support vector machine (SVM) and logistic regression (LR)] were used to construct the diagnostic algorithms. All 13N-glycan structure abundances analyzed by NGFP demonstrated significant and was enrolled by Lasso. Among the three machine learning models, LR algorithm demonstrated the best diagnostic performance for identifying ANHCC in training cohort (LR: AUC: 0.842, 95%CI: 0.784-0.899; RF: AUC: 0.825, 95%CI: 0.766-0.885; SVM: AUC: 0.610, 95%CI: 0.527-0.684). This LR algorithm achieved a high diagnostic performance again in the independent validation (AUC: 0.860, 95%CI: 0.824-0.897). Furthermore, the LR algorithm could stratify ANHCC into two distinct subgroups with high or low risks of overall survival and recurrence in follow-up validation. In conclusion, the biomarker panel consisting of 13N-glycan structures abundances using the best-performing algorithm (LR) was defined and indicative as an effective tool for HCC prediction and prognosis estimate in AFP negative subjects.

Clinical and Contrast-enhanced Ultrasound Characteristics of Epithelioid and Classic Hepatic Angiomyolipoma: Comparison With Alpha-fetoprotein–negative Hepatocellular Carcinoma

Hepatic angiomyolipoma (HAML) comprises epithelioid angiomyolipoma (EAML) and classic hepatic angiomyolipoma (CAML). The imaging appearance of HAML varies widely, and EAML is more easily misdiagnosed as hepatocellular carcinoma (HCC) than as CAML. The clinical and contrast-enhanced ultrasound (CEUS) features of CAML, EAML and HCC with negative alpha-fetoprotein protein expression (HCC[AFP–]) were retrospectively reviewed. The hyper-vascular type was more commonly found in CAML and EAML lesions than in HCC lesions. Most lesions were hyper-enhanced in the arterial phase. CAMLs showed prolonged hyper-enhancement or iso-enhancement during the portal and late phases on CEUS, making them easily distinguishable from HCC(AFP–). Some EAML lesions (41.7%) were hypo-echoic, similar to HCC(AFP–). However, the hypo-enhancement of EAML lesions occurred later than that of HCC(AFP–) lesions. Thus, our findings may be useful in distinguishing among these lesions to improve diagnostic accuracy.

Distinct clinical and genetic characteristics of AFP-negative hepatocellular carcinoma patients infected by HBV or HCV.

e16620 Background: The serum level of alpha-fetoprotein (AFP) is a key biomarker in hepatocellular carcinoma (HCC) screening regardless of the high false-negative rates. Studies have characterized AFP negative HCC patients, however, the differences between HBV and HCV infected AFP negative HCC patients have not been discussed. Methods: The clinical profiles and whole-exome sequencing data of 243 patients were downloaded from the TCGA database. Patients were divided into 4 groups based on their virus infection status and serum AFP measurements as indicated in the table. The mutation profiles and clinical features of the patients were analyzed. Results: In the HBV infected groups, the AFP+ patients carried significantly larger tumors (n = 17, 85.88±11.87 mm) compared to the AFP- patients (n = 13, 49.08±8.933 mm, p = 0.026). The HBV+ AFP+ group exhibited the prevalence of vascular invasion (VI, 76%, 13/17) while less than half of the patients in the HBV+ AFP- group developed VI (46%, 6/13). The WES results identified shared mutations in more than 10% of patients in each group. It appeared that 126 shared mutations were restricted to the HBV+ AFP+ group, they are involved in the regulation of ion transmembrane transport (p &lt; 0.001) and interaction between L1 and Ankyrins (p &lt; 0.0001). Only 51 shared mutations were exclusive to the HBV+ AFP- group, they majorly affect developmental growth (p &lt; 0.001) and skeletal system development (p &lt; 0.01). In contrast, the tumor sizes of HCV+ AFP+ and HCV+ AFP- patients are similar (55.18±7.781 vs 48.19±5.564, p = 0.47), while the VI rate is higher in the HCV+ AFP- group (63%, 17/29, Table). In total, 11 shared mutations were restricted to the HCV+ AFP+ group, several of the genes were involved in the generation of precursor metabolites and energy (p &lt; 0.01) and activation of immune response (p &lt; 0.01). 27 shared mutations were exclusive to the HCV+ AFP- group, the enrichment analysis showed that they are related to the regulation of synapse organization (p &lt; 0.0001) and the regulation of chromosome organization (p &lt; 0.01). Conclusions: HBV and HCV infection were associated with distinct clinical features among the AFP positive and AFP negative patients. The differences were underpinned by the mutations exclusive to each group, and they were involved in distinct molecular and cellular functions. [Table: see text]

Current Status and Perspective Biomarkers in AFP Negative HCC: Towards Screening for and Diagnosing Hepatocellular Carcinoma at an Earlier Stage.

Hepatocellular carcinoma (HCC) is one of the most malignant cancer with high morbidity and mortality which lead to a serious burden to society. AFP (alpha-fetoprotein) is the most widely used serum biomarker to detect HCC worldwide. However, no AFP elevation have been found in many HCC and AFP analysis can't be used to screen HCC in these cases. Currently, many studies have been carried out to find reliable biomarker in diagnosing AFP-negative HCC. Such biomarker would help the diagnosis of AFP-negative HCC, ensuring the timely initiation of treatment. In this review, we highlight the important role of biomarkers that can differentiate AFP-negative HCCs, and discuss their potential clinical applications as biomarkers for the diagnosis of AFP-negative HCC.

Diagnosis value of peripheral blood TEMs percentage in the AFP negative early hepatocellular carcinoma

Objective To evaluate the diagnosis value of the percentage of Tie2-expressing monocytes (TEMs) in CD14+ CD16+ monocytes of peripheral blood from hepatocellular carcinoma (HCC) patients with negative AFP and tumor size≤3 cm. Methods Flow Cytometry (FCM) was used to determine the percentage of TEMs in CD14+ CD16+ monocytes of peripheral blood from patients with HCC (n=82), liver cirrhosis (n=29), chronic hepatitis B (n=28), and healthy controls (n=31). Abbott i2000 microparticle chemiluminescence immunoassay analyzer was used to determine the plasma alpha-fetoprotein (AFP) levels. The difference among multi groups was analyzed by the Kruskal-Wallis H test. Two independent groups were analyzed by the Mann-Whitney U test. The chi-square test was used in the rate comparison. The correlation between TEMs and AFP was analyzed by Spearman rank correlation analysis. Morever, the areas under the receiver operating characteristic curves (ROC-AUC), sensitivity and specificity of TEMs or AFP in differentiating HCC, HCC with AFP negative or tumor size≤3 cm were analyzed. Results The percentage of TEMs in CD14+ CD16+ monocytes of peripheral blood from HCC or HCC with negative AFP or HCC with tumor size≤3 cm was significantly higher than that in patients with liver cirrhosis, chronic hepatitis B and healthy controls(P<0.05). ROC-AUC of TEMs and AFP in the diagnosis of HCC were 0.701 (95% CI 0.626-0.768) and 0.712(95% CI 0.638-0.779)respectively. When the cut-off values of TEMs and AFP were 4.95% and 20 μg/L, the sensitivities of TEMs and AFP were 71.95% and 45.12%, and the specificities of TEMs and AFP were 70.45% and 85.23% . The sensitivity of TEMs in the diagnosis of HCC was significantly higher than that of AFP (χ2=12.16, P=0.000) . The specificity of AFP was significantly higher than that of TEMs (χ2=5.57, P=0.018). There was a highest sensitivity (89.02%) in TEMs/AFP method, and there was a highest specificity (93.18%) in TEMs+ AFP method in the diagnosis of HCC.There was no significant difference between the ROC-AUC for the TEMs and the AFP in the diagnosis of 26 patients with tumor size≤3 cm HCC(0.776 vs 0.645, Z=1.805, P=0.071), TEMs/AFP had the highest sensitivity (84.62%) , while TEMs+ AFP had the highest specificity (93.18%) in the diagnosis of tumor size≤3cm HCC. The ROC-AUC for the TEMs in the diagnosis of 45 patients with AFP negative HCC was 0.739 (95% CI 0.648-0.829). The sensitivity and specificity of TEMs were 80.0% and 70.45% respectively. There was no correlation between the level of plasma AFP and the percentage of TEMs (r=-0.169, P=0.129) determined by Spearmans rank correlation coefficient. Conclusions TEMs is valuable in the diagnosis of HCC with negative AFP and tumor size≤3cm, and the two tests of TEMs and AFP can complement each other in the diagnosis of patients with HCC.(Chin J Lab Med, 2018, 41: 121-131) Key words: Carcinoma, hepatocellular; Liver neoplasms; alpha-Fetoproteins; Receptor, TIE-2; Monocytes

Diagnostic value of glypican-3 in alpha fetoprotein negative hepatocellular carcinoma patients

The prognosis of patients with hepatocellular carcinoma(HCC) is generally very poor with a 5-year survival rate of less than 15% since most of them are diagnosed clinically at their late stage. However, the differential diagnosis between alpha fetoprotein(AFP) negative HCC and cirrhotic nodules is still difficult. To evaluate the diagnostic value of glypican-3(GPC3) in patients with AFP negative hepatitis B related HCC. The liver tissue GPC3 (GPC3L) expression was tested from 426 for surgery and 179 of needle biopsies of hepatitis B related HCC patients using immunohistochemistry staining. Serum GPC3 (GPC3S) and AFP were also measured. Among surgical HCC samples, 80.0% of GPC3L expression was positive, however, in paracarcinomatous and cirrhotic nodules were negative. In needle biopsy tissues, GPC3L positively expression was in 74.9%. The sensitivity of AFP>400 µg/L was 25.4%. The GPC3S >3.5 µg/L was determined as a positive. The area of ROC curve of GPC3S was 0.68(95% CI 0.56-0.79,P<0.05) in all HCC patients,0.81 (95% CI 0.62 -0. 98, P<0.05) in AFP greater or equal to 400 µg/L and 0.64 (95% CI 0.51-0.77, P=0.051) in AFP negative patients. The GPC3S was positive in 48.8% of patients with AFP negative. No difference was observed between GPC3L/GPC3S and serum AFP. GPC3 may be helpful in improving diagnosis of HCC and in differentiating diagnosis between AFP negative HCC and cirrhotic nodules.

Consolidative repeat radiofrequency ablation for alpha-fetoprotein negative hepatocellular carcinoma: does it have a role in local tumor control

Objective To retrospectively evaluate the role of consolidative repeat radiofrequency ablation (CRRFA) based on safety margin (SM) analyses in local tumor control for alpha-fetoprotein (AFP) negative hepatocellular carcinoma (HCC) patients who had been shown to have radiological complete ablation (CA) with radiofrequency ablation (RFA). Methods From July 2002 to July 2009,152 AFP negative HCC patients who were shown to have radiological CA with RFA therapy were retrospectively analyzed. Among them, 110 patients had a SM of less than 1 cm and the other 42 patients had a SM of 1cm or more. Among 110 patients with SM less than 1 cm, fifty nine patients accepted CRRFA within 6 months after the first RFA and 51 did not. From these patients, a narrow SM-CRRFA group (n=41) and a narrow SM-single RFA group (n=37) were enrolled respectively. The wide SM-single RFA group (n= 30) was enrolled from the 42 patients with a SM of 1 cm or more.The LTP (local tumor progression)-free survival rate of the 3 groups were compared with a log-rank test. Results One-, two-, three-, four-, and five-year LTP-free survival rates respectively were 97. 1%, 90.9%, 69.6%, 47.2%, and 33. 0% in the narrow SM-CRRFA patients. 85.9%, 66. 5%,43.5%, 15.8%, and 0. 0%, in the narrow SM-single RFA patients, and were 92.7%, 83.7%,59.3%, 36. 9%, and 9.2% in the wide SM-single RFA patients. There were statistically significant differences (χ2 = 14. 789, P= 0. 001) between the groups. Conclusions An ablation zone with an SM of 1 cm or greater was the most important factor for local control of AFP negative HCC ranging from 3 to 5 cm in diameter. For these patients with a SM of less than 1 cm, CRRFA improved the overall local control outcomes. Key words: Hepatocellular carcinoma; Radiofrequency ablation; Alpha fetoprotein; Safety margin; Local tumor progression

Prognosis evaluation in alpha-fetoprotein negative hepatocellular carcinoma after hepatectomy: Comparison of five staging systems

Aims Alpha-fetoprotein (AFP) loses its potentials in treatment evaluation and prognosis prediction in patients with AFP negative (≤20 ng/ml) hepatocellular carcinoma (HCC). The present study was to identify the risk factors affecting postoperative survival of AFP negative patients and to determine the optimal staging system in predicting the survival of these patients. Methods The data of 306 in total and 98 AFP negative patients amongst were retrospectively reviewed. The risk factors affecting survivals of the patients were identified. And various staging systems were compared, including the sixth tumor node metastasis (TNM) system, Okuda staging, Cancer of the Liver Italian Program (CLIP) score, the Barcelona Clinic Liver Cancer (BCLC) staging system, and the Japan Integrated Staging (JIS) score. Results AFP negative patients tended to have intact tumor capsule and earlier staged tumor by TNM, CLIP and BCLC. The independent risk factors worsening overall survival of AFP negative patients were absence of tumor capsule, Child-Pugh classification B, hepatitis B surface antigen positive and BCLC stage B–C. The risk factors promoting tumor recurrence were tumor size of >3 cm, distribution in two lobes, Okuda stage B–C and BCLC stage B–C. Conclusion Normal AFP level implies earlier staged tumors. BCLC has the strongest potential in prognosis evaluation in AFP negative patients.