LCK inhibition downregulates YAP activity and is therapeutic in patient-derived models of cholangiocarcinoma.

BACKGROUND & AIMS: There is an unmet need to develop novel, effective medical therapies for cholangiocarcinoma (CCA). The Hippo pathway effector, Yes-associated protein (YAP), is oncogenic in CCA, but has historically been difficult to target therapeutically. Recently, we described a novel role for the LCK proto-oncogene, Src family tyrosine kinase (LCK) in activating YAP through tyrosine phosphorylation. This led to the hypothesis that LCK is a viable therapeutic target in CCA via regulation of YAP activity. METHODS: A novel tyrosine kinase inhibitor with relative selectivity for LCK, NTRC 0652-0, was pharmacodynamically profiled in vitro and in CCA cells. A panel of eight CCA patient-derived organoids were characterized and tested for sensitivity to NTRC 0652-0. Two patient-derived xenograft models bearing fibroblast growth factor receptor 2 (FGFR2)-rearrangements were utilized for in vivo assessment of pharmacokinetics, toxicity, and efficacy. RESULTS: NTRC 0652-0 demonstrated selectivity for LCK inhibition in vitro and in CCA cells. LCK inhibition with NTRC 0652-0 led to decreased tyrosine phosphorylation, nuclear localization, and co-transcriptional activity of YAP, and resulted in apoptotic cell death in CCA cell lines. A subset of tested patient-derived organoids demonstrated sensitivity to NTRC 0652-0. CCAs with FGFR2 fusions were identified as a potentially susceptible and clinically relevant genetic subset. In patient-derived xenograft models of FGFR2 fusion-positive CCA, daily oral treatment with NTRC 0652-0 resulted in stable plasma and tumor drug levels, acceptable toxicity, decreased YAP tyrosine phosphorylation, and significantly decreased tumor growth. CONCLUSIONS: A novel LCK inhibitor, NTRC 0652-0, inhibited YAP signaling and demonstrated preclinical efficacy in CCA cell lines, and patient-derived organoid and xenograft models. IMPACT AND IMPLICATIONS: Although aberrant YAP activation is frequently seen in CCA, YAP targeted therapies are not yet clinically available. Herein we show that a novel LCK-selective tyrosine kinase inhibitor (NTRC 0652-0) effectively inhibits YAP tyrosine phosphorylation and cotranscriptional activity and is well tolerated and cytotoxic in multiple preclinical models. The data suggest this approach may be effective in CCA with YAP dependence or FGFR2 fusions, and these findings warrant further investigation in phase I clinical trials.

YAP1 activation and Hippo pathway signaling in the pathogenesis and treatment of intrahepatic cholangiocarcinoma.

Intrahepatic cholangiocarcinoma (iCCA), the second most common primary liver cancer, is a highly lethal epithelial cell malignancy exhibiting features of cholangiocyte differentiation. iCCAs can potentially develop from multiple cell types of origin within liver, including immature or mature cholangiocytes, hepatic stem cells/progenitor cells, and from transdifferentiation of hepatocytes. Understanding the molecular mechanisms and genetic drivers that diversely drive specific cell lineage pathways leading to iCCA has important biological and clinical implications. In this context, activation of the YAP1-TEAD dependent transcription, driven by Hippo-dependent or -independent diverse mechanisms that lead to the stabilization of YAP1 is crucially important to biliary fate commitment in hepatobiliary cancer. In preclinical models, YAP1 activation in hepatocytes or cholangiocytes is sufficient to drive their malignant transformation into iCCA. Moreover, nuclear YAP1/TAZ is highly prevalent in human iCCA irrespective of the varied etiology, and significantly correlates with poor prognosis in iCCA patients. Based on the ubiquitous expression and diverse physiologic roles for YAP1/TAZ in the liver, recent studies have further revealed distinct functions of active YAP1/TAZ in regulating tumor metabolism, as well as the tumor immune microenvironment. In the current review, we discuss our current understanding of the various roles of the Hippo-YAP1 signaling in iCCA pathogenesis, with a specific focus on the roles played by the Hippo-YAP1 pathway in modulating biliary commitment and oncogenicity, iCCA metabolism, and immune microenvironment. We also discuss the therapeutic potential of targeting the YAP1/TAZ-TEAD transcriptional machinery in iCCA, its current limitations, and what future studies are needed to facilitate clinical translation.

Clinical application value of modified invagination for pancreaticojejunostomy in pancreaticoduodenectomy / 中华消化外科杂志

Objective To explore the clinical application value of modified invagination for pancreaticojejunostomy in pancreaticoduodenectomy (PD).Methods The retrospective cohort study was conducted.The clinicopathological data of 39 patients who underwent PD in the Affiliated Hospital of Inner Mongolia Medical University from January 2014 to December 2017 were collected.There were 26 males and 13 females,aged (60±7)years,with a range of 41-75 years.All the 39 patients underwent PD,using Child method to reconstruct digestive tract.Of 39 patients,19 undergoing modified invagination for pancreaticojejunostomy and 20 undergoing mucosa-to-mucosa end-to-side pancreaticojejunostomy were allocated to innovative group and traditional group,respectively.Observation indicators:(1)surgical situations;(2) postoperative complications;(3) follow-up.Follow-up was performed by outpatient examination and telephone interview to detect postoperative tumor recurrence,main pancreatic duct dilatation,survival,and discomfort (abdominal pain,bloating,indigestion,etc.) of patients up to October 2018.Measurement data with normal distribution were represented as Mean±SD,and comparison between groups was analyzed by t test.Measurement data with skewed distribution were represented as M (P25,P75) or M (range),and comparison between groups was analyzed by Mann Whitney U test.Count data were expressed as absolute numbers,and comparison between groups was analyzed by chi-square test or Fisher exact probability.Results (1) Surgical situations:operation time,volume of intraoperative blood loss,cases with soft pancreas or hard pancreas (pancreatic texture),pancreatic duct diameter,time of pancreatic duct removal,cases using somatostatin,and duration of postoperative hospital stay of the innovative group were (342±47) minutes,400 mL (300 mL,400 mL),10,9,3.1 cm (2.9 cm,3.4 cm),37 days (32 days,63 days),17,18 days (15 days,22 days),respectively,versus (392±95)minutes,400 mL (300 mL,525 mL),6,14,3.6 cm (2.6 cm,4.2 cm),43 days (34 days,49 days),18,and 24 days (15 days,27days) of the traditional group;there was no significant difference in the volume of intraoperative blood loss,cases with soft pancreas or hard pancreas (pancreatic texture),pancreatic duct diameter,time of pancreatic duct removal,cases using somatostatin,and duration of postoperative hospital stay between the two groups (Z=-0.775,x2 =2.063,Z=-1.155,Z=-0.295,x2 =0.003,Z=-1.286,P＞0.05);but a significant difference in operation time between the two groups (t =-2.114,P＜0.05).(2) Postoperative complications:6 patients in the innovative group had complications,including 1 of delayed gastric emptying,1 of wound infection,1 of pulmonary infection,1 of acute respiratory failure,1 of perihepatic effusion,and 3 of grade A pancreatic leakage;11 patients in the traditional group had postoperative complications,including 1 of bile leakage,2 of delayed gastric emptying,4 of abdominal infection,4 of wound infection,2 of pulmonary infection,1 of ascites,1 of abdominal hemorrhage,1 of pleural effusion,2 of grade A pancreatic leakage,5 of grade B and C pancreatic leakage;the same patient had multiple complications.There was no significant difference in postoperative complications between the two groups (x2=2.174,P＞0.05),but there was a significant difference in postoperative grade B and C pancreatic leakage between the two groups (P＜O.05).Patients with postoperative complications were improved after symptomatic support treatment,and no patient died during the perioperative period.(3) Follow-up:of the 39 patients,33 (18 in the innovation group and 15 in the traditional group) were followed up for 3-57 months,with a median follow-up time of 17 months.Of the 18 patients receiving follow-up in the innovative group,5 died of tumor recurrence and metastasis,with a survival time of 5-24 months,1 had tumor recurrence at 34 months after operation,1 had main pancreatic duct dilatation and intermittent abdominal pain and abdominal distension,5 had indigestion,1 had back pain,and 5 had good recovery.Of 15 patients receiving follow-up in the traditional group,10 died of tumor recurrence and metastasis,with a survival time of 3-57 months,2 had main pancreatic duct dilatation and intermittent abdominal pain and abdominal distension,2 had indigestion,1 had good recovery.Conclusion Compared with the traditional mucosa-to-mucosa end-to-side pancreaticojejunostomy,modified invagination for pancreaticojejunostomy in the PD is safe and feasible,which can simplify the operation,reduce the requirements for the operator's operation skills,shorten the operation time,and reduce incidence of postoperative grade B and C pancreatic leakage.

Aspartate beta-hydroxylase promotes cholangiocarcinoma progression by modulating RB1 phosphorylation.

Cholangiocarcinoma (CCA) is a highly lethal and aggressive disease. Recently, IDH1/2 mutations have been identified in approximately 20% of CCAs which suggests an involvement of 2-oxoglutarate (2-OG) -dependent dioxygenases in oncogenesis. We investigated if the 2-OG dependent dioxygenase, aspartate beta-hydroxylase (ASPH) was important in tumor development and growth. Immunoassays were used to clarify how ASPH modulates CCA progression by promoting phosphorylation of the retinoblastoma protein (RB1). A xenograft model was employed to determine the role of ASPH on CCA growth. Knockdown of ASPH expression inhibited CCA development and growth by reducing RB1 phosphorylation. Expression of ASPH promoted direct protein interaction between RB1, cyclin-dependent kinases, and cyclins. Treatment with 2-OG-dependent dioxygenase and ASPH inhibitors suppressed the interaction between RB1 and CDK4 as well as RB1 phosphorylation. Knockdown of ASPH expression inhibited CCA progression and RB1 phosphorylation in vivo and they were found to be highly expressed in human CCAs. Knockdown of ASPH expression altered CCA development by modulating RB1 phosphorylation, as one of the major factors regulating the growth of these tumors.

Neurofibroma plexiforme del colédoco: una causa rara de ictericia obstructva. Reporte de caso / Plexiform Neurofibroma in the Common Bile Duct: A Rare Cause of Obstructve Jaundice. Case Report

Paciente masculino de 68 años, con ictericia obstructva quien es llevado a sala de operaciones encontrando una lesión que ocluye el 90% de la luz del tercio medio e inferior del colédoco que es reportada como un neurofbroma plexiforme del colédoco.

Male patent, 68 years old, with obstructve jaundice. Near total obstructon of common bile duct was found during laparotomy, pathology reported a plexiform neurofbroma in the common bile duct.

Extrahepatic biliary cancer: New staging classification.

Tumor staging defines the point in the natural history of the malignancy when the diagnosis is made. The most common staging system for cancer is the tumor, node, metastases classification. Staging of cancers provides useful parameters in the determination of the extent of disease and prognosis. Cholangiocarcinoma are rare and refers to cancers that arise from the biliary epithelium. These tumors can occur anywhere along the biliary tree. These tumors have been previously divided into extrahepatic and intrahepatic lesions. Until recently the extrahepatic bile duct tumors have been considered as a single entity per American Joint Commission on Cancer (AJCC) staging classification. The most recent changes to the AJCC classification of bile duct cancers divide the tumors into two major categories: proximal and distal tumors. This practical classification is based on anatomy and surgical management. High quality cross-sectional computed tomography (CT) and/or magnetic resonance (MR) imaging of the abdomen are essential information to accurately stage this tumors. Imaging plays an important role in diagnosis, localization, staging and optimal management of cholangiocarcinoma. For example, it helps to localize the tumor to either perihilar or distal bile duct, both of which have different management. Further, it helps to accurately stage the disease and identify the presence of significant nodal and distant metastasis, which may preclude surgery. Also, it helps to identify the extent of local invasion, which has a major impact on the management. For example, extensive involvement of hepatic duct reaching up to second-order biliary radicals or major vascular encasement of portal vein or hepatic arteries precludes curative surgery and patient may be managed by palliative therapy. Further, imaging helps to identify any anatomical variations in the hepatic arterial or venous circulation and biliary ductal system, which is vital information for surgical planning. This review presents relevant clinical presentation and imaging acquisition and presentation for the accurate staging classification of bile duct tumors based on the new AJCC criteria. This will be performed with the assistance of anatomical diagrams and representative CT and MR images. The image interpretation must include all relevant imaging information for optimum staging. Detailed recommendations on the items required on the radiology report will be presented.

Markers of bile duct tumors.

Biliary tract carcinomas are relatively rare, representing less than 1% of cancers. However, their incidence has increased in Japan and in industrialized countries like the USA. Biliary tract tumors have a poor prognosis and a high mortality rate because they are usually detected late in the course of the disease; therapeutic treatment options are often limited and of minimal utility. Recent studies have shown the importance of serum and molecular markers in the diagnosis and follow up of biliary tract tumors. This review aims to introduce the main features of the most important serum and molecular markers of biliary tree tumors. Some considerable tumor markers are cancer antigen 125, carbohydrate antigen 19-9, carcinoembryonic antigen, chromogranin A, mucin 1, mucin 5, alpha-fetoprotein, claudins and cytokeratins.

Efficacy of different surgical treatment of 104 patients with hilar cholangiocarcinoma and survival analysis / 中国综合临床

Objective To study the efficacy of surgical treatment in hilar cholangiocarcinoma patients. Methods One hundred and four cases underwent surgical treatment of hilar cholangiocarcinoma were retrospective enrolled in the study from 1998 to 2008, including 45 cases of radical resection, 38 cases of palliative resection, vitro bridge drainage in 21 cases, 93 cases of postoperative patients. Ninty-three patients were followed up for 5 - 67 months, the different procedures of the treatment was summerized. Results Oneyear survival rate of radical resection and palliative resection was 80. 0% ( 32/40 ) and 86. 8% ( 33/38 ),respectively. No significant difference between the two groups (P ＞0. 01 ) were found; 2-year survival rate was 67. 5% (27/40) and 39. 5% ( 15/38), with significant difference between the two groups (P ＜0. 01 ) ;3-year survival rates were 37.5% ( 15/40 ) and 13.2% ( 5/38 ), with significant difference between the two groups (P＜0. 05). Palliative resection had higher 1-year survival rate than in vitro bridge drainage(P ＜ 0.01).Conclusion Surgical treatment of hilar cholangiocarcinoma is the most effective way to prolong the survival time of radical resection, radical surgery had better efficacy than palliative surgery, and palliative surgery is superior to external drainage. In patients of severe jaundice combined with biliary drainage infection, preoperative bridge drainage would improve the safety.

The Role of Radiotherapy in the Treatment of Extrahepatic Bile Duct Carcinoma / 대한치료방사선과학회지

Twenty-seven patients with unresectable extrahepatic bile duct carcinoma (n=21) or with microscopic evidence of tumor rest after aggressive surgery for extrahepatic bile duct carcinoma(n=6) between 1985 and 1990 were given radiotherapy consisting intentionally external radiotherapy and /or intraluminal therapy using Gamma-Med 12i (192-lr) high dose rate (HDR) remote control afterloading system following bile drainage procedures and Gianturco stent insertion. The objectives of this study has been to assess the feasibility and effects on survival of a combination of external radiotherapy and brachytherapy with which we hope to achieve optimal loco-regional control for patients with unresectable extrahepatic bile duct tumors. Sixteen patients were men and deleven were women, and the mean age was 58 years (34-70). 10MV X-ray was used for radiation therapy, with the total dose ranging from 45 Gy to 55 Gy, and intraluminal brachytherapy performed after external radiotherapy, with the dose of total 15 Gy. The minimum follow up was 12 months. Failure were predominantly local-regional, without distant failure. Median survival was 10 months; 2-year actuarial survival rates was 21%. Median survival for common hepatic duct(CHD) cancer was 9 months; for common bile duct (CBD) cancer, was 16 months. And median survival for incomplete surgery/external radiotherapy group and external/intraluminal radiotherapy group was 10 months; for external radiotherapy alone group, was 6 months. Use of chemotherapy and/or hyperthermia were not affected in survival. Therefore, our result is that the survival rates in the group of external/intralumial radiotherapy were comparable with ones in the group of incomplete resection/external radiotherapy, andso we believe that the aggressive local and regional radiotherapy can improve the quality of life and the survival length