Predicting Chronic Myocardial Ischemia Using CCTA-Based Radiomics Machine Learning Nomogram.

BACKGROUND: Coronary computed tomography angiography (CCTA) is a well-established non-invasive diagnostic test for the assessment of coronary artery diseases (CAD). CCTA not only provides information on luminal stenosis but also permits non-invasive assessment and quantitative measurement of stenosis based on radiomics. PURPOSE: This study is aimed to develop and validate a CT-based radiomics machine learning for predicting chronic myocardial ischemia (MIS). METHODS: CCTA and SPECT-myocardial perfusion imaging (MPI) of 154 patients with CAD were retrospectively analyzed and 94 patients were diagnosed with MIS. The patients were randomly divided into two sets: training (n = 107) and test (n = 47). Features were extracted for each CCTA cross-sectional image to identify myocardial segments. Multivariate logistic regression was used to establish a radiomics signature after feature dimension reduction. Finally, the radiomics nomogram was built based on a predictive model of MIS which in turn was constructed by machine learning combined with the clinically related factors. We then validated the model using data from 49 CAD patients and included 18 MIS patients from another medical center. The receiver operating characteristic curve evaluated the diagnostic accuracy of the nomogram based on the training set and was validated by the test and validation set. Decision curve analysis (DCA) was used to validate the clinical practicability of the nomogram. RESULTS: The accuracy of the nomogram for the prediction of MIS in the training, test and validation sets was 0.839, 0.832, and 0.816, respectively. The diagnosis accuracy of the nomogram, signature, and vascular stenosis were 0.824, 0.736 and 0.708, respectively. A significant difference in the number of patients with MIS between the high and low-risk groups was identified based on the nomogram (P < .05). The DCA curve demonstrated that the nomogram was clinically feasible. CONCLUSION: The radiomics nomogram constructed based on the image of CCTA act as a non-invasive tool for predicting MIS that helps to identify high-risk patients with coronary artery disease.

Predicting the progression of Parkinson's disease using conventional MRI and machine learning: An application of radiomic biomarkers in whole-brain white matter.

PURPOSE: This study aimed to develop and validate a radiomics model based on whole-brain white matter and clinical features to predict the progression of Parkinson disease (PD). METHODS: PD patient data from the Parkinson's Progress Markers Initiative (PPMI) database was evaluated. Seventy-two PD patients with disease progression, as measured by the Hoehn-Yahr Scale (HYS) (stage 1-5), and 72 PD patients with stable PD were matched by sex, age, and category of HYS and included in the current study. Each individual's T1 -weighted MRI scans at the baseline timepoint were segmented to isolate whole-brain white matter for radiomics feature extraction. The total dataset was divided into a training and test set according to subject serial number. The size of the training dataset was reduced using the maximum relevance minimum redundancy (mRMR) algorithm to construct a radiomics signature using machine learning. Finally, a joint model was constructed by incorporating the radiomics signature and clinical progression scores. The test data were then used to validate the prediction models, which were evaluated based on discrimination, calibration, and clinical utility. RESULTS: Based on the overall data, the areas under curve (AUCs) of the joint model, signature and Unified Parkinson Disease Rating Scale III PD rating score were 0.836, 0.795, and 0.550, respectively. Furthermore, the sensitivities were 0.805, 0.875, and 0.292, respectively, and the specificities were 0.722, 0.697, and 0.861, respectively. In addition, the predictive accuracy of the model was 0.827, the sensitivity was 0.829 and the specificity was 0.702 for stage-1 PD. For stage-2 PD, the predictive accuracy of the model was 0.854, the sensitivity was 0.960, and the specificity was 0.600. CONCLUSION: Our results provide evidence that conventional structural MRI can predict the progression of PD. This work also supports the use of a simple radiomics signature built from whole-brain white matter features as a useful tool for the assessment and monitoring of PD progression.

Radiomics nomogram based on MRI for predicting white matter hyperintensity progression in elderly adults.

BACKGROUND: White matter hyperintensity (WMH) is widely observed in aging brain and is associated with various diseases. A pragmatic and handy method in the clinic to assess and follow up white matter disease is strongly in need. PURPOSE: To develop and validate a radiomics nomogram for the prediction of WMH progression. STUDY TYPE: Retrospective. POPULATION: Brain images of 193 WMH patients from the Picture Archiving and Communication Systems (PACS) database in the A Medical Center (Zhejiang Provincial People's Hospital). MRI data of 127 WMH patients from the PACS database in the B Medical Center (Zhejiang Lishui People's Hospital) were included for external validation. All of the patients were at least 60 years old. FIELD STRENGTH/SEQUENCE: T1 -fluid attenuated inversion recovery images were acquired using a 3T scanner. ASSESSMENT: WMH was evaluated utilizing the Fazekas scale based on MRI. WMH progression was assessed with a follow-up MRI using a visual rating scale. Three neuroradiologists, who were blinded to the clinical data, assessed the images independently. Moreover, interobserver and intraobserver reproducibility were performed for the regions of interest for segmentation and feature extraction. STATISTICAL TESTS: A receiver operating characteristic (ROC) curve, the area under the curve (AUC) of the ROC was calculated, along with sensitivity and specificity. Also, a Hosmer-Lemeshow test was performed. RESULTS: The AUC of radiomics signature in the primary, internal validation cohort, external validation cohort were 0.886, 0.816, and 0.787, respectively; the specificity were 71.79%, 72.22%, and 81%, respectively; the sensitivity were 92.68%, 87.94% and 78.3%, respectively. The radiomics nomogram in the primary cohort (AUC = 0.899) and the internal validation cohort (AUC = 0.84). The Hosmer-Lemeshow test showed no significant difference between the primary cohort and the internal validation cohort (P > 0.05). The AUC of the radiomics nomogram, radiomics signature, and hyperlipidemia in all patients from the primary and internal validation cohort was 0.878, 0.848, and 0.626, respectively. DATA CONCLUSION: This multicenter study demonstrated the use of a radiomics nomogram in predicting the progression of WMH with elderly adults (an age of at least 60 years) based on conventional MRI. LEVEL OF EVIDENCE: 3 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2020;51:535-546.

ICSNPathway: identify candidate causal SNPs and pathways from genome-wide association study by one analytical framework.

Genome-wide association study (GWAS) is widely utilized to identify genes involved in human complex disease or some other trait. One key challenge for GWAS data interpretation is to identify causal SNPs and provide profound evidence on how they affect the trait. Currently, researches are focusing on identification of candidate causal variants from the most significant SNPs of GWAS, while there is lack of support on biological mechanisms as represented by pathways. Although pathway-based analysis (PBA) has been designed to identify disease-related pathways by analyzing the full list of SNPs from GWAS, it does not emphasize on interpreting causal SNPs. To our knowledge, so far there is no web server available to solve the challenge for GWAS data interpretation within one analytical framework. ICSNPathway is developed to identify candidate causal SNPs and their corresponding candidate causal pathways from GWAS by integrating linkage disequilibrium (LD) analysis, functional SNP annotation and PBA. ICSNPathway provides a feasible solution to bridge the gap between GWAS and disease mechanism study by generating hypothesis of SNP → gene → pathway(s). The ICSNPathway server is freely available at http://icsnpathway.psych.ac.cn/.

Preoperative Prediction of Microsatellite Instability in Rectal Cancer Using Five Machine Learning Algorithms Based on Multiparametric MRI Radiomics.

Objectives: To establish and verify radiomics models based on multiparametric MRI for preoperatively identifying the microsatellite instability (MSI) status of rectal cancer (RC) by comparing different machine learning algorithms. Methods: This retrospective study enrolled 383 (training set, 268; test set, 115) RC patients between January 2017 and June 2022. A total of 4148 radiomics features were extracted from multiparametric MRI, including T2-weighted imaging, T1-weighted imaging, apparent diffusion coefficient, and contrast-enhanced T1-weighted imaging. The analysis of variance, correlation test, univariate logistic analysis, and a gradient-boosting decision tree were used for the dimension reduction. Logistic regression, Bayes, support vector machine (SVM), K-nearest neighbor (KNN), and tree machine learning algorithms were used to build different radiomics models. The relative standard deviation (RSD) and bootstrap method were used to quantify the stability of these five algorithms. Then, predictive performances of different models were assessed using area under curves (AUCs). The performance of the best radiomics model was evaluated using calibration and discrimination. Results: Among these 383 patients, the prevalence of MSI was 14.62% (56/383). The RSD value of logistic regression algorithm was the lowest (4.64%), followed by Bayes (5.44%) and KNN (5.45%), which was significantly better than that of SVM (19.11%) and tree (11.94%) algorithms. The radiomics model based on logistic regression algorithm performed best, with AUCs of 0.827 and 0.739 in the training and test sets, respectively. Conclusions: We developed a radiomics model based on the logistic regression algorithm, which could potentially be used to facilitate the individualized prediction of MSI status in RC patients.

Tracking the evolution of esophageal squamous cell carcinoma under dynamic immune selection by multi-omics sequencing.

Intratumoral heterogeneity (ITH) has been linked to decreased efficacy of clinical treatments. However, although genomic ITH has been characterized in genetic, transcriptomic and epigenetic alterations are hallmarks of esophageal squamous cell carcinoma (ESCC), the extent to which these are heterogeneous in ESCC has not been explored in a unified framework. Further, the extent to which tumor-infiltrated T lymphocytes are directed against cancer cells, but how the immune infiltration acts as a selective force to shape the clonal evolution of ESCC is unclear. In this study, we perform multi-omic sequencing on 186 samples from 36 primary ESCC patients. Through multi-omics analyses, it is discovered that genomic, epigenomic, and transcriptomic ITH are underpinned by ongoing chromosomal instability. Based on the RNA-seq data, we observe diverse levels of immune infiltrate across different tumor sites from the same tumor. We reveal genetic mechanisms of neoantigen evasion under distinct selection pressure from the diverse immune microenvironment. Overall, our work offers an avenue of dissecting the complex contribution of the multi-omics level to the ITH in ESCC and thereby enhances the development of clinical therapy.

i-GSEA4GWAS: a web server for identification of pathways/gene sets associated with traits by applying an improved gene set enrichment analysis to genome-wide association study.

Genome-wide association study (GWAS) is nowadays widely used to identify genes involved in human complex disease. The standard GWAS analysis examines SNPs/genes independently and identifies only a number of the most significant SNPs. It ignores the combined effect of weaker SNPs/genes, which leads to difficulties to explore biological function and mechanism from a systems point of view. Although gene set enrichment analysis (GSEA) has been introduced to GWAS to overcome these limitations by identifying the correlation between pathways/gene sets and traits, the heavy dependence on genotype data, which is not easily available for most published GWAS investigations, has led to limited application of it. In order to perform GSEA on a simple list of GWAS SNP P-values, we implemented GSEA by using SNP label permutation. We further improved GSEA (i-GSEA) by focusing on pathways/gene sets with high proportion of significant genes. To provide researchers an open platform to analyze GWAS data, we developed the i-GSEA4GWAS (improved GSEA for GWAS) web server. i-GSEA4GWAS implements the i-GSEA approach and aims to provide new insights in complex disease studies. i-GSEA4GWAS is freely available at http://gsea4gwas.psych.ac.cn/.

The Growth Factors: Potential Biomarkers and Therapeutic Targets in Kidney Diseases.

Background: Kidney diseases are a prevalent health problem worldwide. Although substantial progress has been made in understanding the pathophysiology of kidney disease, currently there is no satisfactory clinical treatment available to prevent or treat kidney disease. Therefore, strategies to establish early diagnosis, identify the key molecules, and develop novel therapeutic interventions to slow the progression of kidney diseases and reduce their complications are encouraged. Summary: The growth factors play a crucial role in the development of kidney diseases. The altered levels of growth factors are usually detected in circulation and urine in the disease course. A growing body of studies has suggested that growth factors, receptors, and related regulators are promising biomarkers for the diagnosis and/or prognosis and potential therapeutic targets for the treatment of kidney diseases. In this review, we summarize recent advances in the potential applications of growth factors for diagnostic biomarkers and therapeutic targets in kidney diseases and highlight their performances in clinical trials. Key Messages: Most diagnostic and therapeutic strategies targeting growth factors are still far from clinical implementation. The better understanding of growth factor-regulated pathophysiology and the progress of new intervention approaches are expected to facilitate the clinical translation of growth factor-based diagnosis and therapy of kidney diseases.

A novel computed tomography radiomic nomogram for early evaluation of small airway dysfunction development.

The common respiratory abnormality, small airway dysfunction (fSAD), is easily neglected. Its prognostic factors, prevalence, and risk factors are unclear. This study aimed to explore the early detection of fSAD using radiomic analysis of computed tomography (CT) images to predict fSAD progress. The patients were divided into fSAD and non-fSAD groups and divided randomly into a training group (n = 190) and a validation group (n = 82) at a 7:3 ratio. Lung kit software was used for automatic delineation of regions of interest (ROI) on chest CT images. The most valuable imaging features were selected and a radiomic score was established for risk assessment. Multivariate logistic regression analysis showed that age, radiomic score, smoking, and history of asthma were significant predictors of fSAD (P < 0.05). Results suggested that the radiomic nomogram model provides clinicians with useful data and could represent a reliable reference to form fSAD clinical treatment strategies.

JAML promotes acute kidney injury mainly through a macrophage-dependent mechanism.

Although macrophages are undoubtedly attractive therapeutic targets for acute kidney injury (AKI) because of their critical roles in renal inflammation and repair, the underlying mechanisms of macrophage phenotype switching and efferocytosis in the regulation of inflammatory responses during AKI are still largely unclear. The present study elucidated the role of junctional adhesion molecule-like protein (JAML) in the pathogenesis of AKI. We found that JAML was significantly upregulated in kidneys from 2 different murine AKI models including renal ischemia/reperfusion injury (IRI) and cisplatin-induced AKI. By generation of bone marrow chimeric mice, macrophage-specific and tubular cell-specific Jaml conditional knockout mice, we demonstrated JAML promoted AKI mainly via a macrophage-dependent mechanism and found that JAML-mediated macrophage phenotype polarization and efferocytosis is one of the critical signal transduction pathways linking inflammatory responses to AKI. Mechanistically, the effects of JAML on the regulation of macrophages were, at least in part, associated with a macrophage-inducible C-type lectin-dependent mechanism. Collectively, our studies explore for the first time to our knowledge new biological functions of JAML in macrophages and conclude that JAML is an important mediator and biomarker of AKI. Pharmacological targeting of JAML-mediated signaling pathways at multiple levels may provide a novel therapeutic strategy for patients with AKI.

The Updates of Podocyte Lipid Metabolism in Proteinuric Kidney Disease.

BACKGROUND: Podocytes, functionally specialized and terminally differentiated glomerular visceral epithelial cells, are critical for maintaining the structure and function of the glomerular filtration barrier. Podocyte injury is considered as the most important early event contributing to proteinuric kidney diseases such as obesity-related renal disease, diabetic kidney disease, focal segmental glomerulosclerosis, membranous nephropathy, and minimal change disease. Although considerable advances have been made in the understanding of mechanisms that trigger podocyte injury, cell-specific and effective treatments are not clinically available. SUMMARY: Emerging evidence has indicated that the disorder of podocyte lipid metabolism is closely associated with various proteinuric kidney diseases. Excessive lipid accumulation in podocytes leads to cellular dysfunction which is defined as lipotoxicity, a phenomenon characterized by mitochondrial oxidative stress, actin cytoskeleton remodeling, insulin resistance, and inflammatory response that can eventually result in podocyte hypertrophy, detachment, and death. In this review, we summarize recent advances in the understanding of lipids in podocyte biological function and the regulatory mechanisms leading to podocyte lipid accumulation in proteinuric kidney disease. KEY MESSAGES: Targeting podocyte lipid metabolism may represent a novel therapeutic strategy for patients with proteinuric kidney disease.

Radiomic nomogram based on MRI to predict grade of branching type intraductal papillary mucinous neoplasms of the pancreas: a multicenter study.

BACKGROUND: Accurate diagnosis of high-grade branching type intraductal papillary mucinous neoplasms (BD-IPMNs) is challenging in clinical setting. We aimed to construct and validate a nomogram combining clinical characteristics and radiomic features for the preoperative prediction of low and high-grade in BD-IPMNs. METHODS: Two hundred and two patients from three medical centers were enrolled. The high-grade BD-IPMN group comprised patients with high-grade dysplasia and invasive carcinoma in BD-IPMN (n = 50). The training cohort comprised patients from the first medical center (n = 103), and the external independent validation cohorts comprised patients from the second and third medical centers (n = 48 and 51). Within 3 months prior to surgery, all patients were subjected to magnetic resonance examination. The volume of interest was delineated on T1-weighted (T1-w) imaging, T2-weighted (T2-w) imaging, and contrast-enhanced T1-weighted (CET1-w) imaging, respectively, on each tumor slice. Quantitative image features were extracted using MITK software (G.E.). The Mann-Whitney U test or independent-sample t-test, and LASSO regression, were applied for data dimension reduction, after which a radiomic signature was constructed for grade assessment. Based on the training cohort, we developed a combined nomogram model incorporating clinical variables and the radiomic signature. Decision curve analysis (DCA), a receiver operating characteristic curve (ROC), a calibration curve, and the area under the ROC curve (AUC) were used to evaluate the utility of the constructed model based on the external independent validation cohorts. RESULTS: To predict tumor grade, we developed a nine-feature-combined radiomic signature. For the radiomic signature, the AUC values of high-grade disease were 0.836 in the training cohort, 0.811 in external validation cohort 1, and 0.822 in external validation cohort 2. The CA19-9 level and main pancreatic duct size were identified as independent parameters of high-grade of BD-IPMNs using multivariate logistic regression analysis. The CA19-9 level and main pancreatic duct size were then used to construct the radiomic nomogram. Using the radiomic nomogram, the high-grade disease-associated AUC values were 0.903 (training cohort), 0.884 (external validation cohort 1), and 0.876 (external validation cohort 2). The clinical utility of the developed nomogram was verified using the calibration curve and DCA. CONCLUSIONS: The developed radiomic nomogram model could effectively distinguish high-grade patients with BD-IPMNs preoperatively. This preoperative identification might improve treatment methods and promote personalized therapy in patients with BD-IPMNs.

An Integrative Nomogram for Identifying Early-Stage Parkinson's Disease Using Non-motor Symptoms and White Matter-Based Radiomics Biomarkers From Whole-Brain MRI.

Purpose: To develop and validate an integrative nomogram based on white matter (WM) radiomics biomarkers and nonmotor symptoms for the identification of early-stage Parkinson's disease (PD). Methods: The brain magnetic resonance imaging (MRI) and clinical characteristics of 336 subjects, including 168 patients with PD, were collected from the Parkinson's Progress Markers Initiative (PPMI) database. All subjects were randomly divided into training and test sets. According to the baseline MRI scans of patients in the training set, the WM was segmented to extract the radiomic features of each patient and develop radiomics biomarkers, which were then combined with nonmotor symptoms to build an integrative nomogram using machine learning. Finally, the diagnostic accuracy and reliability of the nomogram were evaluated using a receiver operating characteristic curve and test data, respectively. In addition, we investigated 58 patients with atypical PD who had imaging scans without evidence of dopaminergic deficit (SWEDD) to verify whether the nomogram was able to distinguish patients with typical PD from patients with SWEDD. A decision curve analysis was also performed to validate the clinical practicality of the nomogram. Results: The area under the curve values of the integrative nomogram for the training, testing and verification sets were 0.937, 0.922, and 0.836, respectively; the specificity values were 83.8, 88.2, and 91.38%, respectively; and the sensitivity values were 84.6, 82.4, and 70.69%, respectively. A significant difference in the number of patients with PD was observed between the high-risk group and the low-risk group based on the nomogram (P < 0.05). Conclusion: This integrative nomogram is a new potential method to identify patients with early-stage PD.

Development and clinical application of deep learning model for lung nodules screening on CT images.

Lung cancer screening based on low-dose CT (LDCT) has now been widely applied because of its effectiveness and ease of performance. Radiologists who evaluate a large LDCT screening images face enormous challenges, including mechanical repetition and boring work, the easy omission of small nodules, lack of consistent criteria, etc. It requires an efficient method for helping radiologists improve nodule detection accuracy with efficiency and cost-effectiveness. Many novel deep neural network-based systems have demonstrated the potential for use in the proposed technique to detect lung nodules. However, the effectiveness of clinical practice has not been fully recognized or proven. Therefore, the aim of this study to develop and assess a deep learning (DL) algorithm in identifying pulmonary nodules (PNs) on LDCT and investigate the prevalence of the PNs in China. Radiologists and algorithm performance were assessed using the FROC score, ROC-AUC, and average time consumption. Agreement between the reference standard and the DL algorithm in detecting positive nodules was assessed per-study by Bland-Altman analysis. The Lung Nodule Analysis (LUNA) public database was used as the external test. The prevalence of NCPNs was investigated as well as other detailed information regarding the number of pulmonary nodules, their location, and characteristics, as interpreted by two radiologists.

MyBASE: a database for genome polymorphism and gene function studies of Mycobacterium.

BACKGROUND: Mycobacterial pathogens are a major threat to humans. With the increasing availability of functional genomic data, research on mycobacterial pathogenesis and subsequent control strategies will be greatly accelerated. It has been suggested that genome polymorphisms, namely large sequence polymorphisms, can influence the pathogenicity of different mycobacterial strains. However, there is currently no database dedicated to mycobacterial genome polymorphisms with functional interpretations. DESCRIPTION: We have developed a mycobacterial database (MyBASE) housing genome polymorphism data and gene functions to provide the mycobacterial research community with a useful information resource and analysis platform. Whole genome comparison data produced by our lab and the novel genome polymorphisms identified were deposited into MyBASE. Extensive literature review of genome polymorphism data, mainly large sequence polymorphisms (LSPs), operon predictions and curated annotations of virulence and essentiality of mycobacterial genes are unique features of MyBASE. Large-scale genomic data integration from public resources makes MyBASE a comprehensive data warehouse useful for current research. All data is cross-linked and can be graphically viewed via a toolbox in MyBASE. CONCLUSION: As an integrated platform focused on the collection of experimental data from our own lab and published literature, MyBASE will facilitate analysis of genome structure and polymorphisms, which will provide insight into genome evolution. Importantly, the database will also facilitate the comparison of virulence factors among various mycobacterial strains. MyBASE is freely accessible via http://mybase.psych.ac.cn.

Consistency of radiologists in identifying pulmonary nodules based on low-dose computed tomography.

BACKGROUND: To study the consistency of radiologists in identifying pulmonary nodules based on low-dose computed tomography (LDCT), and to analyze factors that affect the consistency. METHODS: A total of 750 LDCT cases were collected randomly from three medical centers. Three experienced chest radiologists independently evaluated and detected the pulmonary nodules on 625 cases of LDCT images. The detected nodules were classified into 3 groups: group I (detected by all radiologists); group II (detected by two radiologists); group III (detected by only one radiologist). The consistency with respect to the image features of individual nodules was assessed. RESULTS: A total of 1,206 nodules were identified by the three radiologists. There were 234 (19.4%) nodules in group I, 377 (31.3%) nodules in group II, and 595 (49.3%) nodules in group III. Logistic regression showed that the size, density, and location of the nodules correlated with the detection of nodules. Nodules sized great than or equal to 4 mm were more consistently identified than nodules sized less than 4 mm. Solid and calcified nodules were more consistently identified than sub-solid nodules. Nodules located in the outer zone were more consistently identified than hilar nodules. CONCLUSIONS: There was considerable inter-reader variability with respect to identification of pulmonary nodules in LDCT. Larger nodules, solid or calcified nodules, and nodules located in the outer zone were more consistently identified.

Correlation of vitamin D receptor gene (ApaI) polymorphism with periodontitis: A meta-analysis of Chinese population.

Many studies have tried to elucidate the connection between vitamin D receptor (VDR) gene (ApaI) polymorphism and periodontitis; however, so far there is no consensus. To further assess the impact of ApaI polymorphism on periodontitis risk, we have conducted a meta-analysis of Chinese population. Relevant literatures were searched according to PubMed and Chinese database in January 2019. The strength of correlation was evaluated by combining odds ratio (ORs) and 95% confidence interval (CIs). Six case-control studies were identified with inclusion criteria, including 734 cases of periodontitis and 687 controls. Based on the overall analysis, the VDR ApaI polymorphism was not due to the risk of periodontitis in all models. Subgroup analysis showed that the risk of periodontitis in North China was significantly reduced. To sum up, the study shows that VDR-ApaI polymorphism may be connected with a lower risk of periodontitis in northern China. It is suggested that inferential studies should be conducted in other ethnic groups.

Diverse modes of clonal evolution in HBV-related hepatocellular carcinoma revealed by single-cell genome sequencing.

Hepatocellular carcinoma (HCC) is a cancer of substantial morphologic, genetic and phenotypic diversity. Yet we do not understand the relationship between intratumor heterogeneity and the associated morphologic/histological characteristics of the tumor. Using single-cell whole-genome sequencing to profile 96 tumor cells (30-36 each) and 15 normal liver cells (5 each), collected from three male patients with HBV-associated HCC, we confirmed that copy number variations occur early in hepatocarcinogenesis but thereafter remain relatively stable throughout tumor progression. Importantly, we showed that specific HCCs can be of monoclonal or polyclonal origins. Tumors with confluent multinodular morphology are the typical polyclonal tumors and display the highest intratumor heterogeneity. In addition to mutational and copy number profiles, we dissected the clonal origins of HCC using HBV-derived foreign genomic markers. In monoclonal HCC, all the tumor single cells exhibit the same HBV integrations, indicating that HBV integration is an early driver event and remains extremely stable during tumor progression. In addition, our results indicated that both models of metastasis, late dissemination and early seeding, have a role in HCC progression. Notably, early intrahepatic spreading of the initiating clone leads to the formation of synchronous multifocal tumors. Meanwhile, we identified a potential driver gene ZNF717 in HCC, which exhibits a high frequency of mutation at both single-cell and population levels, as a tumor suppressor acting through regulating the IL-6/STAT3 pathway. These findings highlight multiple distinct tumor evolutionary mechanisms in HCC, which suggests the need for specific treatment strategies.