bluuebunny/crossref_metadata_2025 huggingface dataset:
Final results of early non-invasive gastric cancer detection study using the serum pepsinogen test method in Croatian patients.
Authors: Trivanovic Dragan , Plestina Stjepko , Honovic Lorena , Dobrilla-Dintinjana Renata , Tanaskovic Jelena Vlasic
Published: May 2019

e15551

Background: Gastric cancer (GC) is the 8th most common cancer cause of cancer deaths in Croatia and 2nd most common cause of cancer deaths worldwide. GC of the intestinal type is usually preceded by a chronic atrophic gastritis which is a precancerous change in the stomach. Methods: A reliable diagnostic tool for the early detection of the GC is essential. Screening programs have led to an improvement of overall 5-y survival rate for GC in Japan and Pepsinogen test method was suggested to reduce mortality from gastric cancer. The gold standard for the diagnosis of GC is the pathohistological study of biopsies obtained during an upper GI endoscopy, an invasive method that is too complicated for use in population screening and patients with comorbidity. We have conducted a prospective single center clinical study over a period of > 2 years to evaluate sensitivity and specificity of reagents, and to determine if these reagents can be part of routine. Inclusion criteria for the study were: signed informed consent, life expectancy > 12, and exclusion criteria were: previous treatments for any malignancy, current usage of IPP or NSAIDs medication, poor ECOG performance status ≥3, known history of H. pylori eradication treatment or gastric surgery. We previously reported preliminary results of PG test method in Croatian population. Here, we present mature data after median follow-up of 26 months. Statistical analyses were performed by using a Mann-Whitney U test, multiple logistic regression and the receiver operating characteristics (ROC) to evaluate the predictive power of biomarkers. Results: Blood samples have been collected from patients with suspicious to GC attending for endoscopy or surgery. We used cut off points to evaluate gastric cancer risk: PGI ≤ 70 and PGI/II ratio ≤ 3.0. Of the 116 patients, 25 patients had GC and 91 had non-malignant pathology on tissue biopsy (like atrophic gastritis). Based on an optimal cut-off value calculated by ROC curve analysis had accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of 86.2%, 60.0%, 93.4%, 65.2%and 89.47%, respectively, for the diagnosis of GC. AUC was 0.767 (95% CI 64.0-89.0). Conclusions: The single use of pepsinogen tests is not sufficient for stomach cancer detection; however, it provides a valuable test for selecting a population that needs further diagnosing. Meanwhile, its high specificity could also help to avoid unnecessary endoscopy, especially in older population or patients with heavy burden of comorbidity. Clinical trial information: 2016-0019-34.
Effect of p-PAQR3Thr32 on PD-L1 expression and immune evasion in gastric cancer.
Authors: Ling Zhi-Qiang
Published: May 2019

e15571

Background: Therapies targeted to the immune checkpoint mediated by PD-1 and PD-L1 show antitumor activity in some solid tumors. We have now examined PD-L1 expression and its regulation in gastric cancer with p-PAQR3Thr32 protein. Methods: The expression of PD-L1 at the protein and mRNA levels in gastric cancer cell lines was examined by flow cytometry, real-time RT-PCR and western blot analysis, respectively. The expression of PD-L1 and p-PAQR3Thr32 protein in 319 surgically resected gastric cancer specimens was evaluated by immunohistochemical analysis. Results: The PD-L1 expression level was higher in gastric cancer cell lines positive for p-PAQR3Thr32 protein induced by glucose starvation than in those negative for the p-PAQR3Thr32 protein. Forced expression of p-PAQR3Thr32 protein in gastric cancer cells markedly increased PD-L1 expression, whereas endogenous PD-L1 expression in p-PAQR3Thr32 protein positive gastric cancer cells was attenuated by treatment with PAQR3 siRNAs. Furthermore, expression of PD-L1 was downregulated by inhibitors of the IRF1 and STAT1 in IFNs-PDL1 signaling pathway in gastric cancer cells positive for p-PAQR3Thr32 protein. At clinical tissue level, the expression level of PD-L1 was positively associated with the presence of p-PAQR3Thr32 protein in gastric cancer specimens. Moreover, the expression level of p-PAQR3Thr32 protein was negatively correlated with CD3, CD8, GZMA (CD8 T cell secretory factor) and positively correlated with CD68 (macrophage marker). Conclusions: Our findings that p-PAQR3Thr32 protein induced by glucose deficiency upregulate PD-L1 by activating IFNs-PDL1 signaling pathway in gastric cancer reveal a direct link between p-PAQR3Thr32 protein and PD-L1 expression. It is suggested that p-PAQR3Thr32 protein may be involved in tumor immunosuppression by inhibiting the proliferation and activity of CD8 T cells in gastric cancer tissues.
The impact of method of recurrence detection on esophageal/gastroesophageal junction (EGJ) cancer outcomes.
Authors: Matloubieh Jubin Eghbali , Licona-Freudensten Alexandra Pilar , Baran Andrea M , Dunne Richard Francis , Hezel Aram F , Noel Marcus Smith , Lada Michal J , Peyre Christian G , Jones Carolyn E , Tejani Mohamedtaki Abdulaziz
Published: May 2019

e15580

Background: Trimodality treatment (tx) with neoadjuvant chemoradiation (CRT) followed by esophagectomy is standard tx for locally advanced EGJ cancer. Post-operatively, there is no strong consensus about role of routine surveillance imaging. At the University of Rochester, patients (pts) have surveillance CT scans every 4-6 months (mos) for the first 2 years post-esophagectomy and every 6-12 mos for the next 3 years. Methods: Pts were identified who underwent esophagectomy for T1-T3 EGJ cancer between January 2011 and December 2015 at our institution. Objectives were to describe the impact of timing and methods of recurrence detection (MoRD) on patient outcomes. Recurrence-free (RFS) and overall survival (OS) were graphed via the Kaplan-Meier method. Results: 138 pts underwent esophagectomy for EGJ cancer: 107 (77.5%) were male, median age was 64, and 116 patients (84.1%) had adenocarcinoma. 112 pts (81.2%) had neoadjuvant CRT. The entire cohort’s median OS was 38.4 mos. 68 pts (49.3%) relapsed with a median RFS of 20.0 mos. Recurrence was detected by routine imaging in 36 pts (52.9%), imaging triggered by symptoms in 27 pts (39.7%), and symptoms alone in 5 pts (7.4%). Post-relapse median OS was 2.3 mos when detected based on symptoms alone, 5.0 mos when detected by imaging triggered by symptoms, and 13.7 mos when detected by routine scans (log-rank p = 0.041). There was no significant association between baseline patient/tumor characteristics or pathologic response and MoRD . 53 patients (77.9%) received salvage/palliative tx with a median of 2 tx (IQR = 1). There was no significant association between MoRD and number of salvage/palliative tx. Conclusions: 49.3% of pts relapsed after esophagectomy for EGJ cancer, consistent with current literature. Almost half of relapses were detected based on symptoms despite routine imaging. Increased OS for pts with relapse detected by routine scans is likely related to lead time bias, but may also be related to increased tx intensity or less aggressive tumors. MoRD did not have a measurable impact on number of lines of post-relapse tx. Prospective randomized trials are needed to determine real benefit of regular surveillance scans among EGJ cancer survivors.
Disassociation of ARID1A with genomic ancestry and prognostic impact in an admixed cohort of Brazilian patients with gastric cancer (GC).
Authors: Freitas Helano C. , Nunes Diana Noronha , Bartelli Thais Fernanda , de Amorim Maria Galli , Araujo Luiza Ferreira , Jesus Victor Hugo Fonseca , Begnami Maria D. , da Silva Israel Tojal , Defelicibus Alexandre , Valieris Renan , Boneti Bianca Salgado , Caires Elizabete Maria Pereira de Andrade , Dias-Neto Emmanuel
Published: May 2019

e15585

Background: Loss of expression of ARID1A, a tumor suppressor gene involved in chromatin remodeling and transcription activation, has been associated with worse prognosis in Asian GC patients (Yang et al. 2016). Mutations in ARID1A have been found in 8-27% of GC, usually leading to gene/protein inactivation. Here we evaluated the possible involvement of ARID1A mutations and clinical characteristics, while considering genomic ancestry and survival, in a cohort of Brazilian GC patients. Methods: We included 112 pts diagnosed with GC and treated at AC Camargo Cancer Center before 2013. The study was approved by local IRB. Genomic DNA was used for capture-based enrichment of a customized gene panel including 99 genes. Libraries were sequenced in the NextSeq 500 platform (Illumina), using paired-end reads (2x75bp). For ancestry inference we used a set of ancestry informative markers, covered by target and off-target reads, described by Elhaik et al. (2014). Results: Median age was 64y (37-91), 63% were male, M:F ratio was 1.73. Most cases were classified as Diffuse (47.3%) followed by Intestinal (41.1%), Mixed (2.7%) and 8.9% were deemed unclassifiable by Lauren´s classification. 22.3% were stage I, 20.5% stage II, 42.9% stage III and 14.3% stage IV. 11.6% were in the GEJ and 80.4% were in corpus/antrum. Five patients were EBV positive (4.5%). Genomic ancestry was as follows: 55.4% European, 27.7% Asian, 8.9% African and 8% were highly admixed ( < 50% of any ancestry). ARID1A was mutated in 19% of cases and showed no association with age at diagnosis (p = 0.21), gender (p = 0.76), tumor location (p = 0.55), staging (p = 0.42), Lauren (p = 0.14) or EBV (p = 0.42). ARID1A had no impact on overall survival (OS) (HR 1.17; 95%CI 0.59-2.31; p = 0.6) or disease-free survival (DFS) (HR 1.24; 95%CI 0.66-2.32; p = 0.5), including the subgroup with Asian genomic background: OS-Asia (HR 1.08; 95%CI 0.31-3.81; p = 0.9), DFS-Asia (HR 1.15; 95%CI 0.32-4.12; p = 0.8). Conclusions: ARID1A is a common driver in GC among Brazilian patients. Unlike in Asians, ARID1A was not prognostic in this Brazilian cohort even in the subgroup with a predominant ( > 50%) Asian genomic ancestry.
Treatment modalities and oncological outcomes in a cohort of patients with advanced gastric cancer in Mexico.
Authors: Valenzuela Ana Karen , Meneses Medina Mónica Isabel , Camargo Vanessa Rosas , Verduzco-Aguirre Haydee Cristina , Hernandez-Felix Jorge Humberto , Huitzil Melendez Fidel David
Published: May 2019

e15590

Background: Gastric cancer (GC) represents the third leading cause of cancer related death in Mexico. Treatment modalities and outcomes have been infrequently reported in our population. Methods: we retrospectively reviewed medical records from all consecutive patients with histologically confirmed metastatic GC treated from January 2005 to December 2015 at our institution. EG junction primaries were excluded. Overall survival (OS) was estimated by Kaplan Meier method from histological diagnosis to dead or last date of follow up and compared by Log-rank test. P value < 0.05 was considered significant. Results: 172 patients were included: 53.5% women, median age 55 years. Lauren’s classification: diffuse 57%, intestinal 20%, 3% mixed and 20% non-specified. Signet ring cells were found in 66% of cases. 37% have T4 tumors. Treatment modalities: 22% none treatment, 12% palliative care, 48% systemic chemotherapy at any time of disease course, 29% initial palliative surgery (derivative o gastrectomy), gastrectomy at any time in 14% (n 24). An R0 resection was achieved in 6/24 of patients. For those patients who received systemic chemotherapy (n 83) objective response rate was 34% and disease control rate 70%. Median OS according to treatment is shown in table. Only 35% of patients received second line chemotherapy and 7% a third line. Conclusions: patients with advanced GC have a dismal prognosis in our center. A third of patients present with terminal disease and are ineligible for oncological treatment. Chemotherapy can be offered to less than a half of patients. Palliative gastrectomy is offered to 12%. Multimodal therapy is associated with the best survival but could be offered to less than 10% of patients. [Table: see text]
Chemotherapy (CT) or chemoradiation (CTRT) in postoperative (simple cholecystectomy) carcinoma gall bladder (GBC) with residual disease, unsuitable for revision surgery.
Authors: Agrawal Sushma , Alam Nawed , Rastogi Neeraj , Das KJ Maria , Lal Punita
Published: May 2019

e15594

Background: Approximately 25% patients of GBC present after SC. Revision surgery is the standard of care, which is possible only in few patients because of delayed presentation ( > 3 months after SC), major vessels encasement by residual lymph-nodes or medical reasons. It is unclear whether these patients should be offered CT /CTRT/ CT followed by consolidation CTRT. The purpose of this analysis is to evaluate whether single modality or dual modality is required in these patients. Methods: Records of GBC patients with SC (non-metastatic based on a staging CECT abdomen) registered between January 2008 - December 2016 were retrieved. Based on the presence and extent of residual disease, they were risk stratified into 3 groups: (1) No residual disease [NRD], (2) LR1: residual disease in GB bed, peri-portal and peri-choledochal lymph-nodes, (3) LR2: residual disease in GB bed, peri-portal and peri-choledochal, coeliac, superior mesenteric or paraaortic lymph-nodes. Patients were either offered CT or CTRT due to lack of any guidelines and according to physician discretion. Those receiving CT were offered 6 cycles of Cisplatin and Gemcitabine. Patients in Consolidation CTRT arm received 3-4 cycles of Cisplatin and Gemcitabine followed by consolidation CTRT (Concurrent Capecitabine daily with RT upto 50.4Gy/28#/5.5 weeks). Response assessment was according to RECIST criteria (CR, PR, SD, PD). Overall survival (OS) was computed by Kaplan Meier method. Results: Out of 300 patients with SC, 87 had the inclusion criteria mentioned above and rest were metastatic. At a median follow-up of 21 months (range 2-129 months), the median OS was not reached (NR) for NRD (n = 17) ( both CT and CTRT), 19 months for LR1 (n = 33) (27 mo with CTRT and 19 mo with CT, p = 0.003) and 14 months for LR2 (14 mo with CTRT and 18 mo with CT, p = 0.29). The median OS was NR, 17 mo, 16 mo and 9 mo for CR (n = 30), PR(n = 31), SD (n = 5), PD (n = 5), p = 0.001. Conclusions: While NRD should be offered single modality, Consolidation CTRT benefits LR1 but not LR2. Risk stratification according to residual disease burden after SC is a prognostic marker for OS.
Development of radiotherapeutic rhenium-188 liposomes in alginate microspheres (Rhe-LAMs) for treatment of liver tumors and technetium-99m liposomes in alginate microspheres (Tech-LAMs) for image-guided treatment planning.
Authors: Bitar Ryan D. , Lopera Jorge , Bao Ande , Merlo Jaclyn , Cigarroa Francisco , Phillips William
Published: May 2019

e15599

Background: Radio-embolic agents such as beta-emitting yttrium-90 spheres have been widely adopted as a modality for liver cancer therapy; however, their production can be timely and costly, shunting to the lungs may occur, and post-procedural visualization is limited. Alginate, a polysaccharide which can easily be formed into microspheres, has already been investigated for drug delivery applications; however, we propose utilizing alginate to manufacture radioembolic microspheres for intra-arterial delivery to liver tumors: Rhenium-188/186-labeled liposomes in alginate microspheres (Rhe-LAMS) as a radioembolic agent for the treatment of liver tumors and technetium-99m labeled liposomes in alginate microspheres (Tech-LAMs) as an agent for nuclear image-guided pre-treatment planning for liver cancer patients. Methods: Liposomes were manufactured and labeled with either Re-188/186 or Tc-99m. The liposomes were then mixed with alginate solution and then cross-linked with CaCl2 to form microspheres. Microsphere diameter was evaluated via light microscopy, and retention of radioactivity was measured over time via dosimeter. Microspheres containing free Re-186/Tc-99m (i.e. no liposomes) were also constructed for control comparison. To test in vivo stability, Tech-LAMs were intra-arterially injected into the liver of rabbits for nuclear imaging. Results: 2 ml batches of Rhe-LAMS/Tech-LAMS of 20-80 microns could be manufactured in 3 hours. Radiolabeling efficiency of the liposomes reached 85% and retention of radioactivity in microspheres reached 75%. After overnight incubation, 90% activity was retained. Control microspheres showed a retention of < 5%. In vivo imaging revealed absent activity in the lungs and high embolic activity in the liver. Conclusions: Our novel method demonstrated success regarding radioactivity retention and embolization capabilities. We envision this method to be a quickly-producible, cost-efficient, and effective means for radioembolization of liver tumors that could be adopted by any radiopharmacy.
Tumor mutation burden in biliary tract cancers with ERBB family or DNA damage repair gene mutations.
Authors: Zhang Yongjie , Shen Ningjia , Yu Wenlong , Fu Xiaohui , Song Jia , Zhao Jing , Zhang Yuzi , Cai Shangli
Published: May 2019

e15602

Background: Biliary tract cancers (BTCs) are aggressive malignancies associated with resistance to chemotherapy. Though immunotherapy presents a promising results based on several studies, the overall response rate is relative low. Tumor mutation burden (TMB) has been considered as a predictive biomarker of immunotherapy. However, the association between common mutations and TMB in BTCs are still unclear. Methods: We analyzed 76 BTC samples (32 cholangiocarcinoma and 44 gallbladder cancer patients) from the Cancer Genome Altas (TCGA) by whole-exome sequencing and 408 samples (276 cholangiocarcinoma and 132 gallbladder cancer patients) from Chinese clinical cohort by 381-gene targeted sequencing. The association between ERBB family members or DNA damage repair (DDR) genes and TMB was explored, respectively. TMB was defined as total number of somatic non-synonymous mutations in coding region. Results: The mutation frequency of ERBB genes was similar between the TCGA cohort and the clinical cohort (cholangiocarcinoma, 13.6% vs 10.9%; gallbladder cancer, 21.9% vs 17.4%). However, DDR mutation rate from the TCGA data was greater than that from the clinical data without consideration the pathological type (cholangiocarcinoma, 40.9% vs 21.0%; gallbladder cancer, 62.5% vs 29.5%). In cholangiocarcinoma, any ERBB gene mutation was associated with higher TMB level in TCGA cohort (P = 0.0013) and clinical cohort (P = 0.0008). Any DDR gene mutation also significantly contributed to higher TMB in two cohorts (TCGA cohort, P = 0.0001; clinical cohort, P = 0.0139). In gallbladder cancer, the same result was discovered on the significant association between ERBB genes statue and higher TMB in both two data (TCGA cohort, P = 0.0370; clinical cohort, P = 0.0008). On the contrary, higher TMB level was observed in patients carrying any DDR alterations (P = 0.0017) in TCGA cohort, but the association was not found in Chinese cohort (P = 0.2660). Besides, prognosis analysis was performed on patients in TCGA cohort. Cholangiocarcinoma patients harboring any ERBB gene mutation represent significantly poorer overall survival (median, 10.6 vs. 40.1 months; HR, 3.37; P = 0.0186). Conclusions: Chinese BTC patients may exhibit distinct gene mutations. These findings were the supplement of better comprehending BTC genomic characteristics. Further studies are needed to evaluate the efficacy of immunotherapy in BTC patients with ERBB or DDR mutation in Chinese clinical cohort.
Derazantinib (DZB) provides antitumor efficacy regardless of line of therapy in patients (pts) with FGFR2-fusion positive advanced intrahepatic cholangiocarcinoma (iCCA).
Authors: Droz Dit Busset Michele , El-Rayes Bassel F. , Harris William Proctor , Damjanov Nevena , Masi Gianluca , Rimassa Lorenza , Bhoori Sherrie , Niger Monica , Personeni Nicola , Braiteh Fadi S. , Lonardi Sara , Braun Stephan , Engelhardt Marc , Saulay Mikael , Schwartz Brian E. , Kazakin Julia , Shaib Walid Labib , Mazzaferro Vincenzo , Papadopoulos Kyriakos P.
Published: May 2019

e15607

Background: FGFR2 fusions are prevalent in 13-22% of iCCA and known oncogenic drivers. DZB is a kinase inhibitor with potent pan-FGFR activity. In a non-comparative Phase 2a study, DZB was administered to 29 pts with FGFR2-fusion positive advanced, inoperable iCCA, either as first-line (1L) (n = 2), 2L (n = 13), 3L (n = 10), 4L (n = 2) or 5L therapy (n = 2). The objective response rate (ORR) with DZB was 21%, disease-control rate (DCR) 83% and median PFS 5.7 months (Mazzaferro et al. 2018 BJC). Data from biliary tract cancer studies suggest decreasing treatment effects of chemotherapy with increasing lines of treatment. Here, we present a post-hoc analysis of outcomes of pts treated with DZB in 1L/2L (n = 15) compared to pts treated post-2L (n = 14). Methods: Pts received 300 mg DZB QD PO. Eligibility criteria included locally confirmed, positive testing of FGFR2 fusion expression (FISH or NGS), ECOG PS 0-1. Objective responses were determined using RECIST 1.1. Disease control rate was defined as CR, PR or SD. Results: The mean age of pts treated in 1L/2L was 66y and 55y in post-2L; 73% were females in 1L/2L and 50% in post-2L treatment; other demographic variables were balanced between groups (87% vs 86% of liver target lesions, median baseline lesion size of 97.5 mm vs 109.5 mm, ECOG PS0 was 60% vs 71%). Of 15 1L/2L group pts, 12 (80%) had prior platinum-based chemotherapy as compared to all 14 pts in the post-2L group. In the 1L/2L and post-2L groups, ORR was 20% and 21%, DCR was 80% and 86%, and a reduction in sum of the largest diameter of target lesions was observed in 60% and 64% of pts, respectively. Median PFS was 5.5 mo (95% CI, 1.9-11.9) and 6.2 mo (3.6-9.2) for the 1L/2L and post-2L groups, respectively. Types of drug-related adverse events were similar in 1L/2L and post-2L. Conclusions: Anti-tumor efficacy of DZB in iCCA patients measured either by ORR, DCR, tumor shrinkage or PFS was numerically similar irrespective of treatment line. These data suggest that DZB is an effective treatment option that can be applied early in the treatment continuum of iCCA patients or at later stages to offer anti-tumor efficacy and disease control. Clinical trial information: 01752920.
The combination of the NT157 and sorafenib as a new therapeutic option for hepatocellular carcinoma.
Authors: Guo Hui , Wang Yu , Yu SZ , Li WY
Published: May 2019

e15610

Background: Sorafenib, which was the only choice of systemic therapy in the past ten years in HCC. But sorafenib alone achieves limited efficacy. Reversing sorafenib resistance was difficulty for oncologist. Based on previous study, we investigated whether NT157 augmented antitumor activity of sorafenib and underlying mechanisms. Methods: HCC cell lines and xenografts were employed to determine antitumor activity of NT157 combined with sorafenib and explore underlying mechanisms. Results: NT157 inhibited proliferation and induced apoptosis in dose and time dependent manner in HCC cells. HCC tumor was efficiently inhibited by NT157 in vivo. Mechanistic investigations elucidated NT157 induced phosphorylation of IRS1 by ERK pathway, and blockade of Stat3 pathway was independent of ERK pathways. NT157-activated p-ERK committed them to degradation by proteasome. CCL20 and CXCL8 were selected as efficacy markers by sequence, which effectively and minimal invasively predict efficacy. No major toxicity of liver, kidney and hemogram was observed, but SII (systemic immune-inflammation = N*P/L) was much lower, implying NT157 may influence immune and inflammatory reaction. Combination of NT157 and sorafenib increased percentage of apoptotic cells, c-caspase3 and PARP. Besides, sorafenib can reverse transient elevation of p-ERK by NT157, which may be potential mechanism of two drugs performing synergy. In vivo tumor volume and weight decreased in combination group compared with monotherapy. NT157 decreased p-AKT, which was overexpressed in resistant cell, implying NT157 may reverse sorafenib resistance by inhibiting AKT. To explore mechanism of sorafenib resistance in HCC, we established sorafenib-resistant HCC cell. Combination of two drugs inhibited cell viability, increased the percentage of apoptotic cells and activated of caspase3 and PARP cleavage in resistant cell. Conclusions: Our findings demonstrate NT157 displays potent antitumor effect in HCC via blockade IRS1 and STAT3 pathways. Combination of NT157 and sorafenib has potential therapeutic effect, even in sorafenib-resistant HCC cell. These findings identify a novel therapeutic strategy for HCC using a combination of NT157 and sorafenib.
Association of the ratio of CD8+ and CD163+ lymphocytes with clinical outcome in hepatocellular carcinoma.
Authors: Chen Huan , Xin Haibei , Wu Lihong , Chen Yanhui , Luo Hongli , Li Nan , Zhang Henghui
Published: May 2019

e15613

Background: Within the tumor microenvironment (TME), infiltrating lymphocytes and myeloid cells including tumor-associated macrophages (TAMs) and tumor-associated neutrophils (TANs) are key players involved in liver cancer progression. The purpose of the study was to explorer whether distinct infiltrated immune cell features differentially affect clinical outcome in hepatocellular carcinoma (HCC). Methods: We obtained respectable stage II HCC specimens, along with adjacent para-tumor tissues from 221 patients who underwent surgical resection at Eastern Hepatobiliary Surgery Hospital, (in Shanghai, China) from 2015 through April 2018. CD8+, CD163+ and CD66B+ tumor-infiltrating lymphocytes (TILs) in the cancer area (CA) and stroma area (SA), as well paratumor stroma area, were analyzed by multiple immunohistochemistry. Results: Hierarchical clustering analysis of immune cell densities revealed that all HCC samples can be classified into three distinct groups. The three immune oncology types (IO-types) were characterized by a strong CD8 T cell density in CA and SA region (IO-1), an intermediate state of CD8 and CD163+ (IO-2), and a strong CD163+ macrophage density in IO-3. Remarkably increasing risks of mortality and recurrence, as well as elevated AST, ALP, GGT and AFP levels, were identified in IO-3 group, when compared with IO-1 group. We then identified that percentages of CA-CD8+ TILs in the tumor sample and SA-CD163+ macrophages in the para-tumor region showed opposite distribution pattern among the three IO types, suggesting a predictive role for CD8/TAM ratio in HCC cohort. Therefore, we next classified all HCC samples into two subgroups, according to the levels of tumor-CA-CD8/paratumor-SA-CD163 ratio. Expectedly, higher rate of CD8/CD163 represented significantly improved overall survival (OS) and progression free survival (PFS), verses lower rate of CD8/CD163. Further association studies suggested that the two subgroups correlated with HBV DNA, tumor size, and microvascular invasion (MVI). Of note, a prognostic signature combining portal vein tumor thrombus (PVTT) and CD8/CD163 ratio discriminated HCC patients into four subtypes with increasing risk of mortality and recurrence. Conclusions: The current results indicated that the CD8/CD163 is a novel, independent prognostic factor for a lower rate of disease recurrence and favorable OS in patients with resectable HCC.
A novel preclinical model of cholangiocarcinoma based on human aberrant FBXW7 expression.
Authors: Wang Jingxiao , Wang Haichuan , Peters Michele , Ding Ning , Ribback Silvia , Utpatel Kirsten , Evert Matthias , Cigliano Antonio , Dombrowski Frank , Song Xinhua , Cossu Antonio , Xu Meng , Che Li , Gordan John Dozier , Calvisi Diego , Chen Xin
Published: May 2019

e15624

Background: Pre-clinical models that mimic human genetic events occurring in intrahepatic cholangiocarcinoma (iCCA) are limited. The ubiquitin ligase F-box and WD repeat domain-containing 7 (FBXW7) is recognized as a tumor suppressor in many cancer types. Methods: Firstly, we determined the FBXW7 mutation frequency (n = 120) and mRNA expression (n = 87) in a collection of human iCCA. Based on the preliminary findings in human data, we generated a mouse model by hydrodynamic tail vein injection of activated/myristylated (myr-)AKT with Fbxw7ΔF, a dominant negative form of Fbxw7. Subsequently, we investigated the role of established targets of Fbxw7, namely Notch2, Yap, and c-Myc in this novel mouse model and in human CCA cell lines. Results: FBXW7 mRNA expression is almost ubiquitously downregulated (71/82; 86.6%) in human iCCA specimens, while only 0.8% of samples showed FBXW7 somatic mutations. In vivo, co-expression of AKT and Fbxw7ΔF triggered the development of iCCA lesions and mice were euthanized by 15 weeks post-injection due to high tumor burden. At the molecular level, a strong induction of FBXW7 canonical targets, including Yap, Notch2, and c-Myc oncoproteins, was detected. However, only c-Myc was consistently confirmed as a FBXW7 target in human CCA cell lines. Interestingly, selected ablation of c-Myc completely impaired iCCA formation in AKT/Fbxw7ΔF mice, whereas deletion of either Yap or Notch2 delayed cholangiocarcinogenesis in the same model. Furthermore, in human iCCA specimens, a strong, inverse correlation between the expression levels of FBXW7 and c-Myc was observed. Conclusions: Downregulation of FBXW7 is almost ubiquitous in human iCCA and cooperates with AKT to induce cholangiocarcinogenesis in mice. This pre-clinical mouse model could be used to test novel therapeutics targeting c-Myc, Notch2, and/or Yap.
A novel subpopulation of circulating tumor cells in patients with cholangiocarcinoma.
Authors: Reduzzi Carolina , Vismara Marta , Silvestri Marco , Celio Luigi , Raimondi Alessandra , Torchio Martina , Pusceddu Sara , Niger Monica , Peverelli Giorgia , De Braud Filippo G. , Daidone Maria Grazia , Cappelletti Vera
Published: May 2019

e15637

Background: In cholangiocarcinoma (CCA), molecular analysis during disease progression is rarely feasible due to low access to tissue. Circulating tumor cells (CTCs) could overcome this issue. Data on CTCs in CCA are presently insufficient to promote their use in clinical management due to their low detection rates. Improved methods to capture all CTC subpopulation are therefore needed. Methods: Peripheral blood (PB) was longitudinally collected from patients with metastatic/unresectable CCA (intrahepatic, extrahepatic and gallbladder). CTCs were enriched from PB (10mL) with Parsortix, labeled for epithelial and leukocyte markers, and analyzed in the DEPArray to collect i) single CTCs positive for epithelial markers (eCTC) and ii) single double negative cells (DNC) lacking epithelial and leukocyte markers. DNCs underwent WGA and low-pass WGS to detect copy number alterations. DNCs with aberrant genotype were defined as non-conventional CTCs (ncCTCs). Results: Forty-one blood samples were collected from 21 patients receiving gemcitabine and/or platinum-based regimens. We detected 18 eCTCs and 73 ncCTCs, respectively in 8/41 (19%, median number = 1.5, range 1-5) and 31/41 (76%, median number = 2, range 1-7) samples. By considering also ncCTCs, CTC-positivity increased by 4.4-fold (83% of CTC+ samples). All untreated patients were CTC+, and CTC- samples (n = 7) were collected only on-treatment. In 18 patients, the detection of eCTCs ( ≥1eCTC/10mL blood) at baseline was associated with a shorter overall survival than in eCTC- patients (7.0 vs 19 months, p = 0.03). CTC count fluctuations during treatment mirrored patient response. Indeed, in responding patients CTCs were not detectable. In some patients, CTC counts were more informative on treatment outcome than imaging and Ca19.9 levels. Conclusions: We developed a new single-cell protocol to detect two distinct CTC subpopulations (eCTC and ncCTC) in patients with CCA. Baseline eCTCs detected by our method inform prognosis whereas ncCTCs provide hints on treatment response. The possibility to collect and molecularly characterize CTC from all patients with our protocol, opens the way to future treatment planning based on CTC molecular asset.
Mutation profiles in circulating tumor DNA (ctDNA) to predict the efficacy of sorafenib treatment in patients with advanced hepatocellular carcinoma (HCC).
Authors: Yang Ping , Ji Guanghui , Jiang Qionghui , Lu Zejun , Yu Yao , Yang Yadong , Jin Ge
Published: May 2019

e15648

Background: To find biomarkers from blood samples through ctDNA and leucocyte genomic DNA sequencing to predict the efficacy of sorafenib. Methods: CtDNA and leucocyte genomic DNA were extracted from patients with advanced hepatocellular carcinoma for sequencing analysis before and after targeted therapy with sorafenib. Mutation profiles were obtained by deep sequencing of 620 selected genes for further bioinformatics analysis. Gene mutation frequency and status were further integrated with clinical data to dynamically monitor and predict the efficacy of sorafenib. Results: Twelve patients with advanced hepatocellular carcinoma admitted to our hospital from March 2017 to September 2018 were enrolled. A total of 27 blood samples were collected for further analysis. The results showed that there were significant differences in BRD4 and FANCG gene mutation abundance between therapeutic effect group and poor effect group (p = 0.014). The mutations of BRD4, FANCG and TP53 were associated with shorter PFS (BRD4 mutation type vs wild type: 12 weeks vs 32 weeks, p = 0.0011; FANCG mutation type vs wild type: 16 weeks vs 32 weeks, p = 0.0103; TP53 mutation type vs wild type: 8 weeks vs 32 weeks, p = 0.0095). Furthermore, we found another 19 genes (SDHA, OTUD7A, TMPRSS2, NAV3, AKT1, EIF2S2, DICER1, MAP3K1, RAD9A, TCF3, IRS1, PIK3C2G, CTCF, TSHZ2, ITPKB, KDM5C, STAG2, AMER1, CDC27) associated with therapeutic efficacy, and dynamic changes of these 19 genes mutation alleys could predict therapeutic efficacy. Conclusions: Our results showed that mutations in ctDNA could predict the efficacy and prognosis of sorafenib for advanced hepatocellular carcinoma, and that dynamic monitoring of mutation frequency in ctDNA of peripheral blood samples could be an additional method to monitor therapeutic efficacy of sorafenib. The clinical significance of the mutations of BRD4, FANCG, TP53 and another 19 genes we found needs to be confirmed on a larger scale.
Chromosome 8q24 amplification predicts prognosis for patients with advanced hepatocellular carcinoma (HCC).
Authors: Cheng Yuan , Huang Mengli , Xie Wenzhuan , Gao Chan , Cai Shangli , Ji Jing , Pan Mingxin
Published: May 2019

e15654

Background: China accounts for over 50% of global liver cancer burden and most patients have advanced disease at the time of diagnosis. Currently, very little is known about the mutational landscape of Chinese HCC patients because it is risky to perform tissue biopsy on late-stage patients. In this study, this problem was overcome by genetic profiling using circulating tumor DNA (ctDNA) and frequently mutated loci were evaluated as biomarkers for prognosis prediction. Methods: Targeted next generation sequencing (NGS) was performed on ctDNA obtained from patients with advanced HCC using a 150-gene panel. The relationship between overall survival (OS) and genetic alterations was analyzed using Kaplan-Meier method. The predictive value of frequently mutated genes was then validated using 607 western HCC cases of the cBioPortal (http://www.cbioportal.org,including TCGA and other two studies). Impact of 8q24 amplification on transcription of other genes was analyzed using mRNA data from TCGA database. Results: Fifty-eight Chinese advanced HCC patients were enrolled and ctDNA was detected in 89.7% of them. The most frequently mutated genes were TP53 (57.7%) and TERT (30.8%) and amplification of chromosome 8q24 occurred at a higher frequency (20.0%) than previously reported for western HCC patients. The 8q24 amplified patients showed significantly shorter OS compared to those without 8q24 amplification (4.9 months vs. Not Reached, p = 0.008). This was confirmed by the cBioPortal database, where 8q24 amplification, which was identified in 10.1% of the HCC cases, was also associated with decreased OS (55.6 months vs.102.7 months, p < 0.001). Gene expression analysis using TCGA mRNA data revealed that upon 8q24 amplification, transcription of 56 genes was upregulated and 169 genes were downregulated, among which ABC transporters, RAS signaling and Hippo signaling pathways were significantly enriched in 8q24 amplified population (p = 0.001, 0.015, 0.027, respectively) as suggested by KEGG analysis. Conclusions: Amplification of chromosome 8q24 ocurred at a higher frequency in Chinese HCC patients and showed promise as a biomarker for prognosis prediction.
Impact on gemcitabine plus cisplatin in unresectable hilar cholangiocarcinoma patients according to effective biliary drainage: A multicenter study.
Authors: Kang Jingu , Lee Jong-Chan , Kim Min Jae , Yang Se Yeol , Woo Sang Myung , Jung Jae Hyup , Hwang Jin-Hyeok
Published: May 2019

e15658

Background: Hilar cholangiocarcinoma (HC) is known to have a poor prognosis due to cholangitis and lack of effective chemotherapeutic regimen. Since 2010, gemcitabine plus cisplatin (GP) is recommended as first-line chemotherapy in cholangiocarcinoma. However, it is still unclear whether GP is effective on HC. We aimed to evaluate the impact on GP in unresectable HC patients according to effective biliary drainage (EBD). Methods: From 2010 to 2016, 112 patients with unresectable HC confirmed by biopsy at three tertiary referral hospitals in Korea were enrolled. 38 patients received GP [cisplatin (25 mg per square meter of body-surface area) followed by gemcitabine (1000 mg per square meter), each administered on days 1 and 8, every 3 weeks]. 74 patients received best supportive care (BSC). The EBD group was defined as a total bilirubin decrease of less than 2 mg/dl within 2 weeks after the first drainage procedure or 50% reduction than pre-procedure total bilirubin. Overall survival (OS) was evaluated in GP group and BSC group according to EBD achieved using the Kaplan–Meier survival curve and the log-rank test. Results: When compared with the BSC group (n = 74), the GP group (n = 38) obtained a benefit in OS (median OS, 6.14 vs 12.7 months, p < 0.001). Moreover, patients with EBD experienced significantly longer OS compared with ineffective biliary drainage (IEBD) patients (median OS, 8.03 vs 5.01 months, p < 0.005). GP chemotherapy was beneficial for OS in IEBD group (p < 0.001) as well as in EBD group (p < 0.005). In the multivariate analysis, GP chemotherapy (HR 0.33, 95% CI 0.21–0.52, p < 0.001), EBD (HR 0.60, 95% CI 0.40–0.91, p < 0.001) and 2 more metastatic site (HR 3.21, 95% CI 1.66–6.18, p < 0.001) were significant prognostic factors. Conclusions: GP chemotherapy provides longer survival in HC, regardless of effective or ineffective biliary drainage.
Integrated multi-omic analysis to reveal comprehensive tumor heterogeneity and novel immunophenotypic classification in primary liver cancer.
Authors: Zhang Qi , Lou Yu , Bai Xueli , Liang Tingbo
Published: May 2019

e15662

Background: Hepatocellular carcinoma (HCC) is heterogeneous,especially in multifocal tumors, which decreases the efficacy of clinical treatments. Understanding tumor heterogeneity is critical when developingnovel treatment strategies. However, a comprehensive investigation of tumor heterogeneity in HCC is lacking, and the available evidence regarding tumor heterogeneity has not ledto improvements inclinical practice. Methods: We harvested 42 samples from eight HCC patients and evaluatedtumor heterogeneity using whole-exome sequencing, RNA sequencing, mass spectrometry-based proteomics and metabolomics, cytometry by time-of-flight, and single-cell analysis. Immunohistochemistry and quantitative polymerase chain reactions (qPCRs) were performed to confirm the expression levels of genes. Results: Tumor heterogeneity is considerable with regard to the genomes, transcriptomes, proteomes, and metabolomes of lesions and tumors. The immune status oftheHCC microenvironment had a relatively low level of heterogeneity. Targeting local immunity could be a suitable intervention with balanced precision and practicability. By clustering immune cells in the HCC microenvironment, we identified three distinctive HCC subtypes with immunocompetent, immunodeficient, and immunosuppressive features. We further revealed the specific metabolic features and cytokine/chemokine expression levels of the different subtypes. Determiningthe expression levels of PTPRCand FOXP3using qPCR and immunohistochemistry in two independent HCC cohorts facilitated the correct classification of HCC patients and the prediction of their prognosis. Conclusions: There is comprehensive intratumoral and intertumoral heterogeneity inall dimensions ofHCC. Based on the results, we propose a novel immunophenotypic classification of HCCs that facilitates prognostic prediction and may support decision making with regard to the choice of therapy.
A retrospective analysis of clinical outcomes among patients with infiltrative hepatocellular carcinoma (iHCC): A single institution study.
Authors: Hannan Lindsay Marie , Chiorean E. Gabriela , Cohen Stacey , Coveler Andrew L. , Shankaran Veena , Wong Kit Man , Mieloszyk Rebecca , Johnson Guy E , Park James Oh , Bhargava Puneet , Harris William Proctor
Published: May 2019

e15667

Background: iHCC is an aggressive disease with poorly characterized treatment outcomes. We sought to describe clinical and radiographic features of iHCC, describe overall survival (OS), and compare outcomes of sorafenib (SOR) use with a cohort of non-infiltrative hepatocellular carcinoma (niHCC) patients. Methods: Radiology reports from the University of Washington with an Li-RADS (LR) assignment, 2013-2017, were reviewed (5037 reports, 1954 patients (pts)) for terms indicative of iHCC. Results: : 102 cases were identified, with 43% arising from known niHCC. iHCC size is difficult to measure but when reported, median was 8cm (interquartile range 5-10cm). Other radiographic features: vascular invasion 72%, R lobe only 62%. Clinical data were obtained via medical record review. The cohort was primarily male (83%), White (65%), age > 55 (88%), HCV (74.5%), heavy alcohol use (67%). At diagnosis: BCLC stage C (76%), ALBI grade 2 (62%) and 3 (21%). Median OS after diagnosis was 5.7 months (m). 38 (37%) pts were too ill to receive therapy. OS for de novo iHCC and for iHCC arising from niHCC did not differ (7.5m vs 5.1m, p = NS). All niHCC pts who received SOR in any line of therapy were identified among pts with a LR 5 lesion in at least one of the aforementioned radiology reports, 2013-2017. Pts who received SOR on trial and/or combined with another agent were excluded. Clinical data for the 83 niHCC pts were abstracted and compared to the 25 iHCC pts who received SOR, any line of therapy. Based on χ2 analyses, the 2 groups did not differ by age, sex, race, BCLC stage at SOR initiation, ECOG status at SOR initiation, HCC etiology, ALBI grade at SOR initiation, or previous locoregional therapy. OS after initiation of SOR was 14.1m for niHCC, 10.5m for iHCC, p = NS. PFS after SOR was 3.7m for niHCC, 7m for iHCC, p = NS. AFP at the time of SOR initiation was higher in iHCC than niHCC (median 1331 vs 48, respectively, p = 0.004), with median %AFP change after 30 days -13% in iHCC and +6% in niHCC, p = NS. Decrease in AFP after 30 days of therapy was associated with improved OS among niHCC (21m versus 7m, p = 0.001) but not among iHCC pts (p = NS). Conclusions: iHCC is a poorly understood subset of HCC with underlying biology not well defined. Compared to niHCC, clinical outcomes with sorafenib do not appear to differ in this cohort. Future characterization of outcomes with Y-90 and immune checkpoint inhibitors is warranted.
Nivolumab for hepatocellular carcinoma (HCC) in a real-world context.
Authors: Rao Nisha , Rizk Emanuelle M. , Hwang Regina , Gartrell Robyn Denise , Makkar Jasnit , Oza Jay , Najjar Marc , Emond Jean , Fox Alyson , Verna Elizabeth , Brejt Sidney , Reis Stephen , Sheynzon Vladimir , Sperling David , Weintraub Joshua , Saenger Yvonne M.
Published: May 2019

e15675

Background: Novel immunotherapies may be effective in patients with HCC; however, patients with advanced liver disease are often excluded from clinical trials. The goal of our study was to evaluate the safety and efficacy of nivolumab for HCC in a real-world context, including among patients with advanced liver disease. Methods: We examined all cases of HCC initiated on off-study nivolumab alone or in combination with locoregional therapy at our institution from 2/2017 to 7/2018. Our sample included 14 patients. We report participant demographics, baseline liver function, treatment toxicity, and overall survival (OS). Results: The majority of participants were male (71%) and Caucasian (57%). NAFLD (36%) and HCV (43%) were the most frequently cited risk factors for HCC. Patients had variable liver disease at baseline, with 36% Child-Turcotte-Pugh (CTP) A, 36% CTP B, and 21% CTP C disease. Patients received either nivolumab only (50%), or nivolumab with locoregional therapy (LRT) (50%). All patients with CTP C disease received nivolumab alone. Treatment was generally safe, with only grade 1 or 2 toxicities reported, most commonly fatigue. Median OS was 7 months. Of note, one patient with baseline CTP C disease showed complete response to nivolumab treatment but experienced recurrence off nivolumab. Conclusions: Our study is the first to report the use of nivolumab in patients with HCC and CTP C disease. Nivolumab was well-tolerated in a real-world context in this small patient cohort, including among patients with advanced cirrhosis. Further study in larger patient populations is warranted. [Table: see text]
Molecular differences between AFP-negative and AFP-positive hepatocellular carcinoma.
Authors: Ling Wu yu , Li Lifeng , Chen Shifu
Published: May 2019

e15681

Background: Alpha-fetoprotein (AFP) is a major biomarker for diagnosis of hepatocellular carcinoma (HCC). Screening of molecular differences between AFP-negative and AFP-positive HCC can be helpful for its diagnosis. Methods: Genomic mutations, gene expressions and clinicopathological data of 270 HCC patients were collected from TCGA database. According to preoperative AFP level, patients were categorized into AFP- (0-20ng/mL, n = 112) and AFP+ ( > 20 ng/mL, n = 158) groups. We identified significantly mutated genes and differentially expressed genes (DEGs) between the two groups and analyzed their correlation with prognosis. Results: The 1-, 3-, and 5-year overall survival (OS) rates were 91.5%, 76.3%, and 67.9% in AFP- and 84.3%, 62.5%, and 34.2% in AFP+. The difference was statistically significant (p = 0.0053). To identify AFP-biased mutation genes, we focused on highly mutated genes in each group (≥5% mutation frequency). At FDR = 0.05, we identified 11 AFP-biased genes: CTNNB1, VWA5B2, HERC2, ZFYVE26, SPEN, MUC6, DST, WDR87, DNAH1, PTPN13, and FANCM. The most striking gene identified was CTNNB1, which is a WNT pathway oncogene. CTNNB1 was more frequently mutated in AFP- than in AFP+ (35.8% vs. 17.0%, p = 0.00059). The OS for HCC was calculated on the basis of the mutation of each gene, which showed that MUC6 mutation was negatively associated with OS (p = 0.018), while other ten genes showed no obvious difference. With the mRNA expression data, a total of 949 DEGs were identified, consisting of 718 upregulated in AFP+, and 231 upregulated in AFP-. The enriched pathways of the DEGs included cell cycle, metabolism of xenobiotics by cytochrome P450 and chemical carcinogenesis. AVP, CCL20 and MUC6 genes were identified as hub genes in the PPI network. Conclusions: According to the results of this study, the AFP level is associated with patients’ OS rates, and AFP- patients tend to have higher CCTNB1 mutation rate. Our ongoing study will verify this result by enrolling new HCC patients.