Extrahepatic Cells Research Articles

Metabolic dysfunction-associated steatotic liver disease-induced changes in the antioxidant system: a review.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a heterogeneous condition characterized by liver steatosis, inflammation, consequent fibrosis, and cirrhosis. Chronic impairment of lipid metabolism is closely related to oxidative stress, leading to cellular lipotoxicity, mitochondrial dysfunction, and endoplasmic reticulum stress. The detrimental effect of oxidative stress is usually accompanied by changes in antioxidant defense mechanisms, with the alterations in antioxidant enzymes expression/activities during MASLD development and progression reported in many clinical and experimental studies. This review will provide a comprehensive overview of the present research on MASLD-induced changes in the catalytic activity and expression of the main antioxidant enzymes (superoxide dismutases, catalase, glutathione peroxidases, glutathione S-transferases, glutathione reductase, NAD(P)H:quinone oxidoreductase) and in the level of non-enzymatic antioxidant glutathione. Furthermore, an overview of the therapeutic effects of vitamin E on antioxidant enzymes during the progression of MASLD will be presented. Generally, at the beginning of MASLD development, the expression/activity of antioxidant enzymes usually increases to protect organisms against the increased production of reactive oxygen species. However, in advanced stage of MASLD, the expression/activity of several antioxidants generally decreases due to damage to hepatic and extrahepatic cells, which further exacerbates the damage. Although the results obtained in patients, in various experimental animal or cell models have been inconsistent, taken together the importance of antioxidant enzymes in MASLD development and progression has been clearly shown.

RAB18 regulates extrahepatic siRNA-mediated gene silencing efficacy

Small interfering RNAs (siRNAs) hold considerable therapeutic potential to selectively silence previously “undruggable” disease-associated targets, offering new opportunities to fight human diseases. This therapeutic strategy, however, is limited by the inability of naked siRNAs to passively diffuse across cellular membranes due to their large molecular size and negative charge. Delivery of siRNAs to liver through conjugation of siRNA to N-acetylgalactosamine (GalNAc) has been a success; providing robust and durable gene knockdown, specifically in hepatocytes. Yet, the poor delivery and silencing efficacy of siRNAs in other cell types has hindered their applications outside the liver. We previously reported that a genome-wide pooled knockout screen identified RAB18 as a major modulator of GalNAc-siRNA conjugates. Herein, we demonstrate RAB18 knockout/knockdown efficaciously enhances siRNA-mediated gene silencing in hepatic, extrahepatic cell lines, and in vivo. Our results reveal a mechanism by which retrograde Golgi-ER transport and the intracellular lipid droplets (LDs) positively regulate siRNA-mediated gene silencing.

64Cu2+ Complexes of Tripodal Amine Ligands' In Vivo Tumor and Liver Uptakes and Intracellular Cu Distribution in the Extrahepatic Bile Duct Carcinoma Cell Line TFK-1: A Basic Comparative Study.

Copper (Cu) is a critical element for cancer cell proliferation and considerably accumulates in the nucleus. 64Cu2+ is an anticancer radiopharmaceutical that targets the copper requirement of cancer cells. However, intravenously injected 64Cu2+ ions primarily accumulate in the liver. Ligand complexation of 64Cu2+ may be a promising method for increasing tumor delivery by reducing liver uptake. In this study, we used three tripodal amine ligands [tris(2-aminoethyl)amine (Tren), diethylenetriamine (Dien), and tris(2-pyridylmethyl)amine (TPMA)] to enclose 64Cu2+ ions and compared their in vivo tumor and liver uptakes using a tumor-bearing xenograft mouse model of the extrahepatic bile duct carcinoma cell line TFK-1. We examined intracellular Cu distribution using microparticle-induced X-ray emission (micro-PIXE) analysis of these compounds. 64Cu2+-Tren and 64Cu2+-Dien showed higher tumor uptake than 64Cu2+-TPMA and 64Cu2+ ions in TFK-1 tumors. Among the three 64Cu2+ complexes and 64Cu2+ ions, liver uptake was inversely correlated with tumor uptake. Micro-PIXE analysis showed that in vitro cellular uptake was similar to in vivo tumor uptake, and nuclear delivery was the highest for 64Cu2+-Tren. Conclusively, an inverse correlation between tumor and liver uptake was observed using three 64Cu2+ complexes of tripodal amine ligands and 64Cu2+ ions. These results provide useful information for the future development of anticancer 64Cu radiopharmaceuticals.

Performance of sofosbuvir and NITD008 in extrahepatic neuronal cells against HEV

Hepatitis E is an underestimated disease, leading to estimated 20 million infections and up to 70,000 deaths annually. Infections are mostly asymptomatic, but can reach mortality rates up to 25% in pregnant women or become chronic in immunocompromised patients. Hepatitis E virus (HEV) infection have been associated with a range of extrahepatic manifestations, including a spectrum of neurological symptoms. Current therapy options are limited to non-specific antivirals like ribavirin, but recently, repurposed viral polymerase inhibitors like sofosbuvir and NITD008 were described to inhibit HEV replication. Here, we evaluated the efficacy of these drugs in various neuronal-derived cell lines to determine their potency outside the liver. Our findings indicate that both drugs, especially sofosbuvir, exhibited reduced efficacy in neuronal cells compared to hepatic cells. These results should be taken into account in the development of direct-acting antivirals for HEV and their potency at extrahepatic replication sites.

The ketone body β-Hydroxybutyrate as a fuel source of chondrosarcoma cells

Chondrosarcoma (CS) is a malignant bone tumor arising from cartilage-producing cells. The conventional subtype of CS typically develops within a dense cartilaginous matrix, creating an environment deficient in oxygen and nutrients, necessitating metabolic adaptation to ensure proliferation under stress conditions. Although ketone bodies (KBs) are oxidized by extrahepatic tissue cells such as the heart and brain, specific cancer cells, including CS cells, can undergo ketolysis. In this study, we found that KBs catabolism is activated in CS cells under nutrition-deprivation conditions. Interestingly, cytosolic β-hydroxybutyrate dehydrogenase 2 (BDH2), rather than mitochondrial BDH1, is expressed in these cells, indicating a specific metabolic adaptation for ketolysis in this bone tumor. The addition of the KB, β-Hydroxybutyrate (β-HB) in serum-starved CS cells re-induced the expression of BDH2, along with the key ketolytic enzyme 3-oxoacid CoA-transferase 1 (OXCT1) and monocarboxylate transporter-1 (MCT1). Additionally, internal β-HB production was quantified in supplied and starved cells, suggesting that CS cells are also capable of ketogenesis alongside ketolysis. These findings unveil a novel metabolic adaptation wherein nutrition-deprived CS cells utilize KBs for energy supply and proliferation.

First report on establishment and characterization of the extrahepatic cholangiocarcinoma sarcoma cell line CBC2T-2

Inflammatory bowel disease (IBD) has as a main characteristic the exacerbation of the immune system against enterocytes, compromising the individual’s intestinal microbiota. This inflammatory cascade causes several nutritional deficiencies, which further compromise immunological functioning and, as a result, worsen the prognosis. This vicious cycle can be interrupted as the patient’s dietary pattern meets their needs according to their clinical condition, acting directly on the inflammatory process of IBD through the interaction of food, intestinal microbiota, and epigenome. Specific nutritional intervention for IBD has a crucial role in preventing and managing disease activity. This review addresses epigenetic modifications through dietary compounds as a mechanism for modulating the intestinal microbiota of patients with IBD.

Distinct immune surveillance in primary biliary cholangitis and primary sclerosing cholangitis is linked with discrete cholangiocarcinoma risk.

Primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) are 2 major liver autoimmune diseases. PBC is common in women and primarily affects intrahepatic small bile duct epithelial cells, known as cholangiocytes. In contrast, PSC is dominant in men and primarily affects medium and big intrahepatic and extrahepatic bile duct epithelial cells. Cholangiocarcinoma (CCA) is a malignancy arising from cholangiocytes, and its incidence is increasing worldwide in both men and women. Numerous retrospective and clinical studies have suggested that PBC patients rarely develop CCA compared to PSC patients. CCA is accountable for the higher deaths in PSC patients due to ineffective therapies and our inability to diagnose the disease at an early stage. Therefore, it is paramount to understand the differences in immune surveillance mechanisms that render PBC patients more resistant while PSC patients are susceptible to CCA development. Here, we review several potential mechanisms contributing to differences in the susceptibility to CCA in PBC versus PSC patients.

Phytochemical research of the anticancer potential of Aloe turkanensis

Abstract. Adem FA, Yenessew A, Yusuf AO, Wanjohi JM. 2022. Phytochemical research of the anticancer potential of Aloe turkanensis. Asian J Nat Prod Biochem 20: 75-88. The number of people diagnosed with cancer is rising worldwide, particularly in Sub-Saharan Africa. The search for new cancer treatments continues to benefit greatly from nature as a rich supply of promising chemicals. The anticancer effects of quinones have made them a popular medicinal class among natural chemicals (e.g., daunomycin and doxorubicin). Aloe turkanensis Christian, like other members of the genus Aloe, is a good place to get quinones. Dried and powdered A. turkanensis rhizomes and leaves were cold percolated in a mixture of dichloromethane and methanol (1:1). The crude extracts significantly decreased the viability of the human extrahepatic bile duct cancer cell line (TFK-1). Twelve chemicals were isolated from the crude extracts through chromatographic separations on silica gel, Sephadex LH-20, and preparative TLC. Spectroscopic techniques such as UV, 1H, and 13C NMR, COSY, NOESY, HMBC, and HSQC were used to determine the structures of the isolated compounds. Two naphthoquinones [3,5,8-trihydroxy-2-methylnaphthalene-1,4-dione (1) and 5,8-dihydroxy-3-methoxy-2-methylnaphthalene-1,4-dione (2)], seven anthraquinones [chrysophanol (3), aloesaponarin I (4), aloesaponarin II (5), laccaic acid D methyl ester (6) helminthosporin (8) aloe-emodin (10) and ?-L-11-O-rhamnopyranosylaloe-emodin (11)], a pre-anthraquinone [aloesaponol I (7)] a pyrone derivative [feralolide (9)] and a benzoic acid derivative [3,4-dihydroxybenzoic acid (12)] were the chemicals that made up these substances. This study reported again that the Aloe genus produced the naphthoquinones [3,5,8-trihydroxy-2-methylnaphthalene-1,4-dione (1) and 5,8-dihydroxy-2-methoxy-2-methylnaphthalene-1,4-dione (2)]. In addition, this study made the first report of 3,5,8-trihydroxy-2-methylnaphthalene-1,4-dione (1) from the Asphodelaceae family. Human extrahepatic bile duct carcinoma (TFK-1) and liver cancer (HuH7) cell lines were used to test the extracted compounds for in-vitro anticancer activity. The anthraquinone aloe-emodin (10) and the naphthoquinone 5,8-dihydroxy-3-methoxy-2-methylnaphthalene-1,4-dione (2) showed significant inhibition against TFK-1 cell lines using the 3- [4,5-dimethylthiazol-2-yl]-2-,5-diphenyltetrazolium bromide (MTT) assay, with IC50 values of 6.0 and Aloesaponol I (7), a pre-anthraquinone, inhibited the development of TFK-1 cells with IC50 values of 10.0 µg/mL and HuH7 cells with 88.0 µg/mL. Both ?-L-11-O- rhamnopyranosyl aloe-emodin (11) and aloesaponarin II (4), an anthraquinone, decreased TKF-1 cell line viability with IC50 values of 23.0 and 34.0 µg/mL, and HuH7 cell line viability with IC50 values of 47.0 and 55.0 µg/mL, respectively. IC50 values of 46.0 µg/mL for helminthosporin (8) showed considerable inhibition of TFK-1 cell growth, although it did not affect HuH7 cells at those concentrations. Extrahepatic bile duct (TFK-1) and liver (HuH7) cancer cell lines are sensitive to these chemicals identified here for the first time. Before these chemicals can be established as potentially effective anticancer medicines, more research on normal cell lines and their mechanism of action is required.

Transmembrane serine protease 2 cleaves nidogen 1 and inhibits extrahepatic liver cancer cell migration and invasion.

We aimed to confirm whether transmembrane serine protease 2 (TMPRSS2) regulates nidogen 1 (NID1) expression in extracellular vesicles (EVs) and metastatic hepatocellular carcinoma (HCC) cells. HCC cells, HUVEC cells, MRC-5 cells, HLE cells, MHCCLM3 cells, MHCC97L cells, H2P cells, H2M cells, as well as LO2 cells were cultured according to providers' instruction and EV models were established by using BALB/cAnN-nu mice to facilitate the verifications. We found that TMPRSS2 expression was inversely correlated with the metastatic potential of HCC cell lines. The expression of TMPRSS2 decreased in a time-dependent manner in tumor-bearing model mice implanted with MHCCLM3 cells compared with uninoculated mice. TMPRSS2 overexpression in MHCCLM3 and MHCC97L cells led to the significant downregulation of NID1 expression in total cell lysates and isolated EVs. In contrast, TMPRSS2 silencing resulted in the elevation of NID1 expression in cells and EVs. Administration of EVs from MHCCLM3 and MHCC97L cells with overexpressed or silenced TMPRSS2 inhibited or strengthened, respectively, the invasion, proliferation, and migration of LO2 tumor cells. EVs derived from MHCCLM3 and MHCC97L cells with overexpressed or depleted TMPRSS2 also deactivated or activated fibroblasts, respectively. These EVs secrete inflammatory cytokines and phosphorylated p65, facilitate the colonization of fibroblasts, and augment fibroblast growth and motility. These findings provide evidence for a new candidate drug targeting tumorigenic EV-NID1 to treat HCC.

Ursodeoxycholic acid inhibits epithelial-mesenchymal transition, suppressing invasiveness of bile duct cancer cells: An in vitro study.

Epithelial-mesenchymal transition (EMT) features are associated with pathological severity in the progression and metastasis of various cancer types, including bile duct cancer (BDC). Our previous study demonstrated that ursodeoxycholic acid (UDCA) blocked the EGFR-MAPK signaling pathway and inhibited the invasion of BDC cells. The present study was performed to determine whether UDCA inhibits EMT and promotes the expression of E-cadherin to inhibit the invasion and aggressiveness of BDC. In addition, the present study aimed to confirm that the primary mechanism of inhibition of EMT by UDCA is related to the EGFR axis. Human extrahepatic BDC cells were cultured. The effect of UDCA on cell proliferation was evaluated using MTT assays. A cell death ELISA kit was used to measure apoptosis, and western blot assays or immunofluorescence staining assays measured the expression levels of various target proteins. The mRNA expression of Slug and ZEB1 was evaluated via reverse transcription-quantitative PCR. The invasiveness of BDC cells was estimated by invasion assays and western blot assays for focal adhesion kinase (FAK). UDCA inhibited the proliferation of BDC cells as effectively as gefitinib (an EGFR inhibitor), and the combination of UDCA and gefitinib revealed an additive effect on the proliferation of cells. UDCA and gefitinib induced apoptosis, and the combination of UDCA and gefitinib demonstrated an additive effect on apoptosis in BDC cells. UDCA restored the E-cadherin expression inhibited by EGF and suppressed N-cadherin expression increased by EGF as effectively as gefitinib. UDCA suppressed the Slug and ZEB1 mRNA expression induced by EGF in BDC cells. UDCA suppressed the invasiveness of BDC cells and FAK expression linked to the invasiveness of BDC. In conclusion, UDCA enhanced E-cadherin expression and suppressed N-cadherin expression through inhibition of the EGF-EGFR axis, contributing to the inhibition of EMT and invasiveness in BDC cells. Therefore, UDCA may be applied as an adjuvant or palliative antineoplastic agent and as a therapeutic option to enhance the effect of other chemotherapeutics.

S1760 Metastatic Cervical Squamous Cell Carcinoma of the Distal Bile Duct

Introduction: Squamous cell carcinoma (SCC) of the bile duct is sporadic. Most cases are primary. Only 2 cases of cervical SCC metastatic to the bile ducts have been reported. Consideration of this diagnosis is essential in patients who have a history of malignancy and present with symptoms of biliary obstruction. Case Description/Methods: A 63-year-old female presented with persistent abdominal pain, nausea, and vomiting. Mild abdominal tenderness and scleral icterus were noted on physical examination. Bilirubin, AST, and ALT were elevated. Magnetic resonance cholangiopancreatography (MRCP) revealed dilated intra- and extra-hepatic bile ducts; however, there was no evidence of an apparent mass or gallstones. A lesion was observed at the ampullary level measuring around 1 cm that was brushed and biopsied on endoscopic cholangiopancreatography (ERCP). A metal stent was placed for decompression in the common bile duct. Pathology revealed poorly differentiated SCC with an immune profile favoring metastasis from the gynecologic tract. The patient was diagnosed with SCC of the cervix 15 months prior to presentation. A biopsy of the cervical tissue showed poorly differentiated cervical SCC with positivity in CK7 and HMWK, scattered positive staining of vimentin. Chemo-radiation was initiated with Cisplatin, Paclitaxel, and Bevacizumab. Next-generation sequencing (NGS) showed an immunohistochemical profile consistent with gynecological cancers, namely, MLH1, PMS2, MSH2, and MSH6. Pembrolizumab was also started due to positive PDL-1 expression and mutations in ATM and KMT2D. These agents were continued after she underwent the biliary decompression. Discussion: Metastatic SCC of the biliary tract is rare. Due to its low incidence, there is no standardized treatment strategy; however, there are reported cases of surgical resection in addition to chemoradiotherapy. Very few cases of cervical SCC have been reported to spread to gastrointestinal organs. NGS is a modality that has been widely implemented. The combination of immunohistology staining and NGS can aid in the diagnosis and help guide chemotherapy. The genetic variations present a valuable tool to aim the treatment of cervical cancer. While survival rates for patients with cervical carcinoma have improved, treatment remains difficult as not enough cases exist to compare treatment modalities. Limited knowledge is present regarding managing extra-hepatic duct squamous cell carcinoma since it is rare.

HOXA5 inhibits the proliferation of extrahepatic cholangiocarcinoma cells by enhancing MXD1 expression and activating the p53 pathway

Homeobox A5 (HOXA5) is a transcription factor in mammalian and can regulate cell differentiation, proliferation, and apoptosis as well as tumorigenesis. However, little is known on whether and how HOXA5 can regulate the malignant behaviors of cholangiocarcinoma. The methylation levels of HOXA5 were evaluated by methylation microarray and bisulfite sequencing PCR. HOXA5 expression in tissue samples was examined by immunohistochemistry and Western blot. The proliferation of tumor cells was assessed by CCK-8, EdU, and nude mouse tumorigenicity assays. The invasion, apoptosis and cell cycling of tumor cells were evaluated by Wound healing assay and flow cytometry. The interaction between HOXA5 and the MXD1 promoter was examined by CUT & Tag assay, luciferase reporter assay and chromatin immunoprecipitation. Hypermethylation in the HOXA5 promoter down-regulated HOXA5 expression in extrahepatic cholangiocarcinoma (ECCA) tissues, which was correlated with worse overall survival. HOXA5 overexpression significantly inhibited the proliferation and tumor growth. HOXA5 overexpression enhanced MXD1 expression by directly binding to the MXD1 promoter in ECCA cells. MXD1 overexpression inhibited the proliferation and tumor growth while MXD1 silencing abrogated the HOXA5-mediated proliferation inhibition. HOXA5 overexpression increased p53 protein expression in an MXD1-dependent manner. HOXA5 and MXD1 acted as tumor suppressors to inhibit the mitosis of ECCA cells by enhancing the p53 signaling. Our findings may uncover molecular mechanisms by which the HOXA5/MXD1 axis regulates the progression of ECCA, suggesting that the HOXA5/MXD1 may be therapeutic targets for ECCA.

Hepatic, Extrahepatic and Extracellular Vesicle Cytochrome P450 2E1 in Alcohol and Acetaminophen-Mediated Adverse Interactions and Potential Treatment Options.

Alcohol and several therapeutic drugs, including acetaminophen, are metabolized by cytochrome P450 2E1 (CYP2E1) into toxic compounds. At low levels, these compounds are not detrimental, but higher sustained levels of these compounds can lead to life-long problems such as cytotoxicity, organ damage, and cancer. Furthermore, CYP2E1 can facilitate or enhance the effects of alcohol-drug and drug-drug interactions. In this review, we discuss the role of CYP2E1 in the metabolism of alcohol and drugs (with emphasis on acetaminophen), mediating injury/toxicities, and drug-drug/alcohol-drug interactions. Next, we discuss various compounds and various nutraceuticals that can reduce or prevent alcohol/drug-induced toxicity. Additionally, we highlight experimental outcomes of alcohol/drug-induced toxicity and potential treatment strategies. Finally, we cover the role and implications of extracellular vesicles (EVs) containing CYP2E1 in hepatic and extrahepatic cells and provide perspectives on the clinical relevance of EVs containing CYP2E1 in intracellular and intercellular communications leading to drug-drug and alcohol-drug interactions. Furthermore, we provide our perspectives on CYP2E1 as a druggable target using nutraceuticals and the use of EVs for targeted drug delivery in extrahepatic and hepatic cells, especially to treat cellular toxicity.

Inhibition of PCSK9 affects serum lipoprotein functions in patients with familial hypercholesterolemia

Background and Aims : PCSK9, beyond regulating plasma cholesterol levels, exerts several pleiotropic effects modulating lipid metabolism in extrahepatic cells, including macrophages. Macrophage cholesterol homeostasis strictly depends on serum lipoprotein functions, including the HDL capacity to promote cell cholesterol efflux (cholesterol efflux capacity, CEC) and the serum capacity to promote cell cholesterol accumulation (cholesterol loading capacity, CLC). The aim of the present study was to evaluate potential changes in HDL-CEC and serum CLC after six-month of treatment with PCSK9 inhibitors in Familial Hypercholesterolemia (FH) subjects.

Gastrointestinal endoscopy in the diagnosis and treatment of biliary liver disease

Biliary liver disease refers to intrahepatic and extrahepatic bile duct system lesions or bile duct epithelial cells damage, resulting in abnormal liver function, inflammation and fibrosis, which mainly manifests as chronic cholestatic liver disease. The common causes include primary biliary cholangitis, primary sclerosing cholangitis, progressive familial intrahepatic cholangitis, bile duct complications after liver transplantation, and vanishing bile duct syndrome caused by drugs or serious infections, and the most prominent symptoms affecting the patients quality life is pruritus and jaundice. Endoscopic retrograde duodenoscopic cholangiopancreatography and cholangiopancreatoscopy (SpyGlass) and other endoscopic examinations can significantly improve the accuracy of early differential diagnosis,cytology and biopsy of primary sclerosing cholangitis, immunoglobulin G4-related cholangitis and cholangiocarcinoma. Endoscopic nasobiliary drainage, balloon dilatation and intrabiliary stent implantation can significantly improve pruritus symptoms, liver biochemical indicators and prognosis. Therefore, gastrointestinal endoscopy has important value and application prospects in the diagnosis and treatment of biliary liver diseases.

Effects of PCSK9 inhibitors on HDL cholesterol efflux and serum cholesterol loading capacity in familial hypercholesterolemia subjects: a multi-lipid-center real-world evaluation.

Proprotein convertase subtilisin/kexin type 9 (PCSK9), beyond regulating LDL cholesterol (LDL-c) plasma levels, exerts several pleiotropic effects by modulating lipid metabolism in extrahepatic cells such as macrophages. Macrophage cholesterol homeostasis depends on serum lipoprotein functions, including the HDL capacity to promote cell cholesterol efflux (CEC) and the serum capacity to promote cell cholesterol loading (CLC). The aim of this observational study was to investigate the effect of PCSK9 inhibitors (PCSK9-i) treatment on HDL-CEC and serum CLC in patients with familial hypercholesterolemia (FH). 31 genetically confirmed FH patients were recruited. Blood was collected and serum isolated at baseline and after 6 months of PCSK9-i treatment. HDL-CEC was evaluated through the main pathways with a radioisotopic cell-based assay. Serum CLC was assessed fluorimetrically in human THP-1 monocyte-derived macrophages. After treatment with PCSK9-i, total cholesterol and LDL-c significantly decreased (−41.6%, p < 0.0001 and −56.7%, p < 0.0001, respectively). Total HDL-CEC was not different between patients before and after treatment. Conversely, despite no changes in HDL-c levels between the groups, ABCG1 HDL-CEC significantly increased after treatment (+22.2%, p < 0.0001) as well as HDL-CEC by aqueous diffusion (+7.8%, p = 0.0008). Only a trend towards reduction of ABCA1 HDL-CEC was observed after treatment. PCSK9-i significantly decreased serum CLC (−6.6%, p = 0.0272). This effect was only partly related to the reduction of LDL-c levels. In conclusion, PCSK9-i treatment significantly increased HDL-CEC through ABCG1 and aqueous diffusion pathways and reduced the serum CLC in FH patients. The favorable effect of PCSK9-i on functional lipid profile could contribute to the cardiovascular benefit of these drugs in FH patients.

Disparities in palliative care utilization in cholangiocarcinoma patients: A National Cancer Database (NCDB) study.

e24077 Background: Cholangiocarcinoma is an adenocarcinoma of the intra- and extrahepatic bile duct epithelial cells. Although cases of cholangiocarcinoma in the US are rare, there is a low 5-year survival rate for these patients. The preferred treatment option for all subtypes is surgical resection. Alternative options include palliative care (PC) treatments such as chemotherapy, radiotherapy and locoregional treatments. There are many obstacles in accurately diagnosing cholangiocarcinoma, including obtaining adequate tumor samples for histology and determining what is malignant vs. benign in patients with primary sclerosing cholangitis (an important risk factor of cholangiocarcinoma). Such delays in diagnosis can lead to delays in treatment, which in turn results in a poor prognosis and worse outcome for patients. To the authors’ knowledge, there is no current study assessing PC disparities in patients with cholangiocarcinoma. Methods: A total of 46,751 patients who had a diagnosis with cholangiocarcinoma were identified utilizing the National Cancer Database (NCDB) ICD-O-3 histology code 8160. The 5-year survival of PC use was analyzed via a Kaplan-Meier curve. Demographic factors (race, biological sex, income, facility type, insurance, geographic location, and Charlson-Deyo co-morbidity score) were studied in relation to the receipt of PC. Results: Approximately 15.16% of our cohort received PC (7,089/46,751). There was a statistically significant difference in the staging of those patients that received palliative care, with 52% of our cohort being stage IV (p &lt; 0.001). The sex of patients who received PC was equally distributed between men and women (51.0% to 49.0%). The majority of PC recipients were white (83.6%) individuals making under $63,000 (69.6%) with a reported Charlson-Deyo score of 0 (64.9%). Many patients who received PC had Medicare (57.6%) and were treated at either an academic/research program (45.4%) or at a comprehensive community cancer program (33.6%). Facility locations were predominantly located in the Mid-South Atlantic and East North Central regions, compared to the Pacific regions (56.3% vs 8.4%), and patients who received PC lived mainly in urban areas (98.2%). The median survival for PC patients was shorter compared to patients that did not receive PC (5.32 months vs 9.23 months). Conclusions: This is one of the most comprehensive studies that identifies the demographic disparities of patients who received PC for cholangiocarcinoma. The typical PC patient is a white individual with Medicare, living in an urban area in the Mid-South Atlantic and East North Central region making under $63,000. By highlighting the disparities that exist, our study can aid clinicians in addressing PC under-utilization in specific demographics to help provide more comprehensive care.

P-104 Overexpression of mir-224-3p in extrahepatic cholangiocarcinoma associated with increased dose of I131 radiotherapy: An in vitro study

Cholangiocarcinoma (CCA) affects the biliary tract, constituting a heterogeneous tumor, being the second most common type of primary liver cancer, a rare tumor type, representing approximately 3% of gastrointestinal neoplasms. Metabolic radiotherapy with Iodine-131 (131 I) is associated with cell death by apoptosis in CCA. The exposure of cells to hypoxia causes genetic and biological changes, allowing adaptation to the microenvironment. In this process, there are a series of genes regulated by hypoxia involved in multiple biological and pathological processes, such as proliferation, metabolism, angiogenesis and cell migration. In this context, MicroRNAs (miRNAs) play important roles in carcinogenesis, acting as oncogenes or tumor suppressors, in this case, the hypoxia-induced transcription factor (HIF), whose expression in primary tumor cell lines is regulated by miR-224-3p. There is also reference to its association with hepatocarcinoma. To analyze the effect of 131 I radiotherapy on miR-224-3p expression in intra (HuCCT-1) and extra-hepatic (TFK-1) CCA cell lines. Human cell lines of CCA (TFK-1 and HuCCT-1) and cholangiocytes (H69) were cultured and submitted to irradiation with 131 I at different doses (1, 20 and 60 Gy) after 2h, 48h and 12 days. Analysis of the expression of miRNAs was performed by real-time polymerase chain reaction. An alpha error of 5% was admitted. In HuCCT-1 cells, underexpression of miR-224-3p was noted under all conditions analyzed relative to control. It was also observed an increase in the expression of miR-224-3p when using the dose of 60Gy in 2h (0.31), 48h (0.39) and 12 days (0.61; P=0.001). In TFK-1 cells, overexpression of miR-224-3p was observed associated with increased dose after 2h (1Gy= 2.28, 20Gy= 3.78 and 60Gy= 5.89; P=0.001). 48 hours after irradiation with 131 I, the expression levels of this miRNA continued to be overexpressed, however, a decrease was observed at all doses (1Gy= 1.53, 20Gy= 1.41 and 60Gy= 2.91; P=0, 0001) compared to cells treated at 2 hours. miR-224-3p, a tumor suppressor in several carcinogenic pathways, has overexpression associated with an increased dose of 131 I radiotherapy in extrahepatic CCA cells.

Verteporfin synergizes the efficacy of anti-PD-1 in cholangiocarcinoma

BackgroundCholangiocarcinoma (CCA) is one of the primary hepatobiliary malignant neoplasms with only 10% of 5-year survival rate. Promising immunotherapy with the blockade of immune checkpoints has no clear benefit in CCA. The inhibition of YAP1 signaling by verteporfin has shown encouraging results by inhibiting cell proliferation and inducing apoptosis. This study aimed to evaluate the potential benefit of the combination of verteporfin and anti-programmed cell death 1 (PD-1) in CCA mouse model. MethodsWe assessed the cytotoxicity of verteporfin in human CCA cell lines in vitro, including both intrahepatic CCA and extrahepatic CCA cells. We examined the in vitro effect of verteporfin on cell proliferation, apoptosis, and stemness. We evaluated the in vivo efficacy of verteporfin, anti-PD-1, and a combination of both in subcutaneous CCA mouse model. ResultsOur study showed that verteporfin reduced tumor cell growth and enhanced apoptosis of human CCA tumor cells in vitro in a dose-dependent fashion. Nevertheless, verteporfin impaired stemness evidenced by reduced spheroid formation and colony formation, decreased numbers of cells with aldehyde dehydrogenase activity and positive cancer stem cell markers (all P < 0.05). The combination of verteporfin and anti-PD-1 reduced tumor burden in CCA subcutaneous SB1 tumor model compared to either agent alone. ConclusionsVerteporfin exhibits antitumor effects in both intrahepatic and extrahepatic CCA cell lines and the combination with anti-PD-1 inhibited tumor growth.

SAHA induce hippo pathway in CCA cells without increasing cell proliferation.

Cholangiocarcinoma is a malignant tumor originating from bile duct epithelial cells. Since tumor metastasis is associated with poor prognosis and short-term survival of patients, there is an urgent need for alternative therapeutic approaches for CCA. Because of that reason, we aimed to investigate effect of SAHA which is known as HDAC inhibitor on extrahepatic cholangiocarcinoma cell line (TFK-1). Cell cycle was measured by Muse Cell Analyzer. YAP, TAZ, TGF-β protein levels were determined by western-blotting method. TEAD (1-3), TIMP2 and TIMP3 genes level were determined by real-time PCR analysis. We have seen the positive effects of SAHA on the TFK-1 cell line as it reduces cell viability and arresting cells in the G0/G1 phase. We also observed the negative effects of SAHA, as it increases the expression levels of YAP, TAZ, TGF-β protein and TEAD (1-3) gene. We also found that SAHA reduced the expression levels of TIMP2 and TIMP3 in TFK-1 cells, but was not statistically significant. Although observing its antiproliferative effects, these negative effects may be related to the cells being resistant to the drug or the remaining cells having a more aggressive phenotype. Therefore, we think that caution should be exercised in the use of this drug for CCA treatment.