Genomic characterization of a large panel of patient-derived hepatocellular carcinoma xenograft tumor models for preclinical development

Lack of clinically relevant tumor models dramatically hampers development of effective therapies for hepatocellular carcinoma (HCC). Establishment of patient-derived xenograft (PDX) models that faithfully recapitulate the genetic and phenotypic features of HCC becomes important. In this study, we first established a cohort of 65 stable PDX models of HCC from corresponding Chinese patients. Then we showed that the histology and gene expression patterns of PDX models were highly consistent between xenografts and case-matched original tumors. Genetic alterations, including mutations and DNA copy number alterations (CNAs), of the xenografts correlated well with the published data of HCC patient specimens. Furthermore, differential responses to sorafenib, the standard-of-care agent, in randomly chosen xenografts were unveiled. Finally, in the models expressing high levels of FGFR1 gene according to the genomic data, FGFR1 inhibitor lenvatinib showed greater efficacy than sorafenib. Taken together, our data indicate that PDX models resemble histopathological and genomic characteristics of clinical HCC tumors, as well as recapitulate the differential responses of HCC patients to the standard-of-care treatment. Overall, this large collection of PDX models becomes a clinically relevant platform for drug screening, biomarker discovery and translational research in preclinical setting.     

Establishment of patient-derived xenograft models of adenoid cystic carcinoma to assess pre-clinical efficacy of combination therapy of a PI3K inhibitor and retinoic acid

Due to the difficulties and long periods of establishment, preclinical animal models of adenoid cystic carcinoma (ACC) are scarce but imperative. The researches involving molecular features and therapeutic targets of ACC require an integrated group of preclinical animal models which can credibly retain the heterogeneity of this tumor. Currently chemotherapies and targeting therapies have modest efficacy in ACC and the overall response rate is rather low. Therefore, novel therapeutic regimen of ACC is urgently needed and remains a major clinical challenge. We transplanted a group of tumor samples from human salivary ACC into immunodeficient mice to establish patient-derived xenografts (PDXs). Patient tumors and their matched PDXs were conducted histological analyses, whole-exome sequencing (WES) and RNA-seq respectively. 13 PDXs were successfully established from 34 ACC, involved in 3 histological types, including cribriform, tubular, and solid. These ACC PDXs generally reflected the histopathological and molecular features of their corresponding original tumors. MYB/MYBL1-NFIB fusion (53.85%) and high-frequency mutation genes, such as KDM6A, KMT2C, KMT2D, NOTCH1, NOTCH2, SMARCA4 and PIK3CA were mainly conserved in PDXs. Guided by the genetic alterations, the efficiencies of retinoic acid (RA) and a PI3K inhibitor were evaluated in ACC PDX models harboring both MYB fusion and PIK3CA amplification/mutation. Combination treatment of the PI3K inhibitor and RA demonstrated remarkable inhibition of tumors in PDXs harboring both PIK3CA mutation/amplification and MYB-NFIB fusion gene in vivo and in vitro. In this study, we displayed the morphologically and genetic featured PDXs which recapitulated the heterogeneity of original ACC tumors, indicating that the models could be used as a platform for drug screening for therapy response. The feasibility of combination treatment approaches for dual targets were confirmed, providing new regimens for personalized therapies in ACC.     

索拉非尼治疗肝细胞癌的疗效评价及不良反应

目的：观察索拉非尼治疗肝细胞癌的临床疗效及不良反应。方法：回顾性分析我院2008年7月至2009年12月间应用索拉非尼400mg每日两次治疗21例肝细胞癌患者的临床疗效与不良反应。结果：2例由于在服药后1月内出现肝功能异常而停药,未进行肿瘤评价。19例接受过1次以上实体瘤治疗反应评价标准（RECIST Response Evaluation Criteria in Solid Tumours 1.0版）的评价,其中无客观缓解者,14例（73.7%）疾病稳定且稳定时间大于3月。5例（26.3%）持续肿瘤进展。药物不良反应最常见的为手足皮肤反应、高血压、腹泻,其次有脱发、乏力、纳差、口腔溃疡、肝功能异常、发热等,不良反应一般在服药后1-2周内出现,其中有2例不能耐受不良反应而减少药物剂量,2例由于肝功能异常而停药。结论：索拉非尼治疗肝细胞癌患者以疾病稳定者多见,应严密监测并及时处理不良反应,以保证持续用药。     

索拉非尼治疗肝细胞癌的疗效评价及不良反应

目的:观察索拉非尼治疗肝细胞癌的临床疗效及不良反应。方法:回顾性分析我院2008年7月至2009年12月间应用索拉非尼400mg每日两次治疗21例肝细胞癌患者的临床疗效与不良反应。结果:2例由于在服药后1月内出现肝功能异常而停药,未进行肿瘤评价。19例接受过1次以上实体瘤治疗反应评价标准(RECIST Response Evaluation Criteria in Solid Tumours 1.0版)的评价,其中无客观缓解者,14例(73.7%)疾病稳定且稳定时间大于3月。5例(26.3%)持续肿瘤进展。药物不良反应最常见的为手足皮肤反应、高血压、腹泻,其次有脱发、乏力、纳差、口腔溃疡、肝功能异常、发热等,不良反应一般在服药后1-2周内出现,其中有2例不能耐受不良反应而减少药物剂量,2例由于肝功能异常而停药。结论:索拉非尼治疗肝细胞癌患者以疾病稳定者多见,应严密监测并及时处理不良反应,以保证持续用药。     

索拉非尼治疗中晚期肝细胞癌的不良反应及其处理

背景与目的：SHARP（sorafenib hcc assessment randomized protocol）和ORIENTAL（sorafenib in patients in asia-pacific region with hepatocellular carcinoma）两个Ⅲ期临床试验证实,多激酶抑制剂索拉非尼在一定程度上能延长晚期肝癌患者的总生存期（overall survival,OS）,并显著改善无进展生存期（progress free survival,PFS）和延长疾病进展时间（time to progression,TTP）,但治疗过程中出现的各种不良反应在一定程度上会影响其临床治疗。本研究使用索拉非尼治疗25例中晚期肝细胞癌患者,并对其不良反应情况及临床处理进行总结。方法：选取2008年1月-2009年10月期间符合原发性肝癌临床诊断标准的晚期肝癌患者25例,给予索拉非尼治疗所有患者均满足以下标准：⑴化疗栓塞术后病情进展;⑵广泛门脉癌栓无法栓塞;⑶肝门部、腹膜后淋巴结或肺、骨等多发转移者;⑷弥漫乏血供肿瘤;⑸签署知情同意书。在开始服用索拉非尼治疗后的前12周,观察患者不同级别不良反应发生情况,并给予相应临床处理。结果：共25例患者接受索拉非尼治疗,其中男性17例,女性8例,年龄范围30～72岁,平均51.44岁。期间9例患者死亡,其中3例在服药开始的12周内死亡。3例患者中断索拉非尼治疗,其中2例为治疗不满12周停药,1例为服药5个月后停药。共20例患者符合观察标准。不良反应发生情况如下：手足皮肤反应（hand-foot-skin reaction,HFSR）4例（4/20）,腹泻4例（4/20）,脱发5例（5/20）,皮疹4例（4/20）,乏力8例（8/20）,白细胞和血小板减少4例（4/20）,高血压1例（1/20）,腹痛1例（1/20）。这些不良反应经临床处理后,20例患者均能继续接受索拉非尼治疗。结论：索拉非尼治疗中晚期肝细胞癌具有良好的安全性和良好的耐受性。     

索拉非尼对中晚期肝细胞癌患者的影响

目的：观察中晚期肝细胞癌患者索拉非尼治疗后药物相关性不良反应的发生率,并评价服药后6个月的疗效及对患者生活质量的影响。方法：对36例服用索拉非尼的肝细胞癌患者所发生的药物相关性不良反应进行分级、记录与统计分析。按照RECIST标准评价服药6个月后的疗效。EORTC QLQ-C30量表评估索拉非尼治疗前与治疗后6个月患者的生活质量。结果：患者服用索拉非尼后出现手足皮肤反应、腹泻、皮疹、高血压等不良反应。患者服药6个月后总反应率为75％。经索拉非尼治疗后患者在躯体、角色、认知、情绪及社会功能评分有所上升,有统计学差异（ P＜0.05）。除腹泻外其他症状评分均有下降,有统计学差异（ P     

Pro‐angiogenic TIE‐2‐expressing monocytes/TEMs as a biomarker of the effect of sorafenib in patients with advanced hepatocellular carcinoma

Sorafenib, a multi‐kinase inhibitor, inhibits tumor angiogenesis and is the first‐line systemic therapy for patients with advanced hepatocellular carcinoma (HCC). However, due to its limited effects and frequent occurrence of side effects, biomarkers are needed to predict the effects of sorafenib. We considered the possibility of using TIE‐2‐expressing monocytes (TEMs) to predict the response in sorafenib‐treated patients with advanced HCC. TEMs serve as a diagnostic marker of HCC and are related to angiogenesis. We analyzed 25 advanced HCC patients and prospectively evaluated TEMs before (Pre TEMs) and at 1 month after initial therapy (T1m TEMs). The radiologic response was evaluated by modified Response Evaluation Criteria in Solid Tumors (mRECIST). Median survival time (MST) was significantly longer in the partial response/stable disease (PR/SD) group (21.8 months) than in the PD group (8.7 months). ΔTEMs (changes of T1m TEMs compared to Pre TEMs) were significantly lower in the PR/SD group than in the PD group. MST of the ΔTEMs low group (14.2 months) was significantly longer than that of the high group (8.7 months). Univariate and multivariate Cox regression analyses showed that ΔTEMs [hazard ratio (HR) = 8.53, 95% confidence interval (CI) = 1.51–48.16, p = 0.015] and Child‐Pugh class (HR = 5.59, 95% CI = 1.06–29.63, p = 0.043) were independently associated with overall survival. Our results suggest that ΔTEMs could serve as a biomarker for predicting radiologic response and overall survival in sorafenib‐treated patients with advanced HCC.     

microRNA-622 acts as a tumor suppressor in hepatocellular carcinoma

microRNAs (miRNAs) are important regulators of tumor development and progression. In this study, we aimed to explore the expression and role of miR-622 in hepatocellular carcinoma (HCC). We found that miR-622 was significantly downregulated in human HCC specimens compared to adjacent noncancerous liver tissues. miR-622 downregulation was significantly associated with aggressive parameters and poor prognosis in HCC. Enforced expression of miR-622 significantly decreased the proliferation and colony formation and induced apoptosis of HCC cells. In vivo studies demonstrated that miR-622 overexpression retarded the growth of HCC xenograft tumors. Bioinformatic analysis and luciferase reporter assays revealed that miR-622 directly targeted the 3′-untranslated region (UTR) of mitogen-activated protein 4 kinase 4 (MAP4K4) mRNA. Ectopic expression of miR-622 led to a significant reduction of MAP4K4 expression in HCC cells and xenograft tumors. Overexpression of MAP4K4 partially restored cell proliferation and colony formation and reversed the induction of apoptosis in miR-622-overexpressing HCC cells. Inhibition of JNK and NF-κB signaling phenocopied the anticancer effects of miR-622 on HCC cells. Taken together, miR-622 acts as a tumor suppressor in HCC and restoration of miR-622 may provide therapeutic benefits in the treatment of HCC.     

甲胎蛋白对索拉菲尼治疗进展期肝细胞癌的预后评价

近几年来以索拉菲尼为代表的分子靶向药物已成为进展期肝细胞癌综合治疗的热点之一,但目前对于索拉菲尼治疗进展期肝细胞癌的预后评价指标尚没有统一的标准。甲胎蛋白作为重要的肝癌肿瘤标志物,已经广泛地应用于肝细胞癌的筛查诊断,并被认为具有潜在的预测肝细胞癌患者的预后价值。本文总结了近几年来关于甲胎蛋白对索拉菲尼治疗进展期肝细胞癌预后评价的相关文献。从总体上看,甲胎蛋白对于索拉菲尼治疗进展期肝细胞癌预后评价作用是值得肯定的,并可以作为传统的基于影像学资料的肝细胞癌预后评价标准的补充,具有广泛的研究前景和应用价值。     

Adverse events of sorafenib in hepatocellular carcinoma treatment

Sorafenib is an oral multikinase inhibitor approved by the US Food and Drug Administration for treatment of the patients with surgically unresectable hepatocellular carcinoma (HCC). Sorafenib mitigates angiogenesis by targeting vascular endothelial growth factor receptors and platelet-derived growth factor receptors in endothelial cells and pericytes. Moreover, it suppresses cell proliferation via blockage of B-RAF and RAF1 of the mitogen-activated protein kinase pathway in tumor cells. Sorafenib has been the standard molecular targeted medication in the treatment of advanced-stage HCC patients ineligible for potentially curative interventional (radiofrequency or microwave ablation) or palliative trans-arterial chemoembolization (TACE) therapies for over a decade. However, it only increases overall survival by less than 3 months, and systemic exposure to sorafenib causes clinically significant toxicities (about 50% of patients). Given the high frequency and severity of these toxicities, sorafenib dose must be often reduced or discontinued altogether. In this review, we discussed the mechanism of sorafenib-associated adverse events and their management during HCC treatment.     

索拉菲尼治疗肝癌常见不良反应及处理的研究进展

索拉菲尼是一种口服多激酶抑制剂。通过作用于Raf激酶直接抑制肿瘤细胞增殖,还可作用于血管内皮生长因子受体1,2,3（VEGFR-1, -2, -3）,以及血小板源生长因子受体-β（PDGFR-β）、受体酪氨酸激酶、抑制肿瘤新生血管生成。索拉菲尼通过抑制肿瘤细胞增殖和抗血管生成的双重作用,从而达到抗肿瘤的目的。已被多个国家批准作为首个系统治疗肝细胞肝癌的分子靶向药物。其常见不良反应包括皮肤反应、恶心、腹泻、体质量减轻、高血压等,影响了患者的长期使用依从性,进而影响治疗效果。正确地认识和管理索拉非尼的不良反应则有助于发挥索拉非尼的治疗作用,提高临床效果。本文从索拉菲尼靶向治疗的常见不良反应、发生机制及处理方法等方面进行综述。      

Development of sorafenib and other molecularly targeted agents in hepatocellular carcinoma

It is well appreciated that hepatocellular carcinoma (HCC) represents one of the most challenging malignancies of worldwide importance. In fact, HCC is the fifth most common cancer and the third most common cause of cancer-related death globally. The incidence rates for HCC in the U.S. and Western Europe have been rising. Unresectable or metastatic HCC carries a poor prognosis, and systemic therapy with cytotoxic agents provides marginal benefit. Because of the poor track record of systemic therapy in HCC, there has been a sense of nihilism for this disease in the oncology community for decades. However, with the arrival of newly developed, molecularly targeted agents and the success of some of these agents in other traditionally challenging cancers, such as renal cell carcinoma, there has been renewed interest in developing novel systemic therapy in HCC. At the recent Annual Meeting of the American Society of Clinical Oncology, results of a phase 3, randomized, placebo-controlled trial were presented in which sorafenib demonstrated improved survival in patients with advanced HCC. This landmark study represents the first agent that has demonstrated an improved overall survival benefit in this disease and sets the new standard for first-line treatment of advanced HCC. For this review, the author concisely summarized the current status of molecularly targeted agents that are under clinical development in advanced HCC.     

经肝动脉化疗栓塞联合索拉非尼治疗巨大肝癌的临床疗效及安全性观察

目的:探讨经肝动脉化疗栓塞（TACE）联合索拉非尼治疗巨大肝癌的临床疗效及安全性。方法:回顾性分析2009年6月-2013年6月我院收治的85例巨大肝癌（直径≥10cm）患者的临床资料。其中采用TACE联合索拉非尼治疗的患者共26例作为观察组,单纯采用TACE治疗的患者共37例作为对照组1,单纯采用索拉非尼治疗的患者共22例作为对照组2。Kaplan-Meier法绘制三组患者的生存曲线,采用Log-rank比较三组的中位生存期、生存率、中位肿瘤进展时间,观察并对比三组的不良反应发生率。结果:观察组、对照组1、对照组2中位生存期分别为16.3个月（95%CI:10.1～22.3个月）、10.4个月（95%CI:7.2～13.5个月）、6.2个月（95%CI:4.2～5.6个月）,差异具有统计学意义（P<0.05）;观察组1年、2年、3年生存率明显高于其他两组,差异具有统计学意义（P<0.05）;观察组、对照组1、对照组2的中位肿瘤进展时间分别为6.5个月（95%CI:3.2～10.4个月）、3.2个月（95%CI:1.6～5.5个月）、2.3个月（95%CI:1.2～3.4个月）,差异具有统计学意义（P<0.05）。三组的不良反应发生率比较差异无统计学意义（P>0.05）。结论:TACE联合索拉非尼治疗巨大肝癌可以延缓肿瘤进展,延长生存期,安全性和耐受性良好,具有良好的临床应用前景。     

Sorafenib and DE605, a novel c-Met inhibitor,synergistically suppress hepatocellular carcinoma

Sorafenib, an oral multikinase inhibitor of Raf, VEGF and PDGF receptor signaling is approved for advanced hepatocellular carcinoma (HCC). One strategy to improve HCC therapy is to combine agents that target key signaling pathways. Aberrant mesenchymal-epithelial transition factor (c-Met) activation is associated with a variety of human malignancies and therefore represents a target for therapy. In this study, we investigated a novel c-Met inhibitor, DE605, together with sorafenib in hepatocellular carcinoma cells in vitro and in vivo. DE605 and sorafenib synergistically induced apoptosis in hepatocellular carcinoma cells. Mechanistically, DE605 activated the FGFR3/Erk pathway, which in turn was inhibited by sorafenib, resulting in synergism. Finally, DE605 and sorafenib significantly inhibited growth of PLC/PRF/5 hepatocellular carcinoma tumor xenografts in athymic nude mice. Importantly, no obvious weight loss (toxicity) was detected. Thus in combination, DE605 and sorafenib target complementary anti-apoptotic pathways and synergistically suppress HCC, providing the rationale for clinical studies with this novel combination.     

Bilirubin inhibits the anticancer activity of sorafenib by blocking MCL-1 degradation in hepatocellular carcinoma cells

Objective: Sorafenib is a first-line drug for advanced hepatocellular carcinoma(HCC). Unfortunately, most patients with HCC do not respond to sorafenib, mainly because of the frequent development of drug resistance. Bilirubin is an end metabolite of heme catabolism and an indicator of liver function, but its direct role in regulating the anticancer activity of sorafenib in HCC cells is unclear. In the current study, we aimed to investigate the mechanism of action of bilirubin in sorafenib-mediat...     

索拉非尼治疗肝细胞癌机制的研究进展

索拉非尼（多吉美,Sorafenib）是唯一被批准用于治疗晚期原发性肝细胞癌(hepatocellular carcinoma,HCC)的口服多激酶抑制剂。随着索拉非尼临床应用的发展,其抗癌机制被越来越多的人关注。索拉非尼不仅能作用于 RAF/MEK/ERK 通路、shp 1/STAT3 相关通路、Wnt/β-catenin 通路和内质网途径,还能影响相关基因和酶类,从而达到抑制肿瘤细胞增殖和肿瘤新生血管生成的作用。本文就近年来索拉非尼治疗 HCC 机制的研究进展作一综述。     

CLDN3 inhibits cancer aggressiveness via Wnt-EMT signaling and is a potential prognostic biomarker for hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the most common fatal malignancies but the molecular genetic basis of this disease remains unclear. By using genome-wide methylation profiling analysis, we identified CLDN3 as an epigenetically regulated gene in cancer. Here, we investigated its function and clinical relevance in human HCC. CLDN3 downregulation occurred in 87/114 (76.3%) of primary HCCs, where it was correlated significantly with shorter survival of HCC patients (P=0.021). Moreover, multivariate cyclooxygenase regression analysis showed that CLDN3 was an independent prognostic factor for overall survival (P=0.014). Absent expression of CLDN3 was also detected in 67% of HCC cell lines, which was significantly associated with its promoter hypermethylation. Ectopic expression of CLDN3 in HCC cells could inhibit cell motility, cell invasiveness, and tumor formation in nude mice. Mechanistic investigations suggested through downregulation of GSK3B, CTNNB1, SNAI2, and CDH2, CLDN3 could significantly suppress metastasis by inactivating the Wnt/β-catenin-epithelial mesenchymal transition (EMT) axis in HCC cells. Collectively, our findings demonstrated that CLDN3 is an epigenetically silenced metastasis suppressor gene in HCC. A better understanding of the molecular mechanism of CLDN3 in inhibiting liver cancer cell metastasis may lead to a more effective management of HCC patients with the inactivation of CLDN3.