Oncopig bladder cancer cells recapitulate human bladder cancer treatment responses in vitro.

Introduction: Bladder cancer is a common neoplasia of the urinary tract that holds the highest cost of lifelong treatment per patient, highlighting the need for a continuous search for new therapies for the disease. Current bladder cancer models are either imperfect in their ability to translate results to clinical practice (mouse models), or rare and not inducible (canine models). Swine models are an attractive alternative to model the disease due to their similarities with humans on several levels. The Oncopig Cancer Model has been shown to develop tumors that closely resemble human tumors. However, urothelial carcinoma has not yet been studied in this platform. Methods: We aimed to develop novel Oncopig bladder cancer cell line (BCCL) and investigate whether these urothelial swine cells mimic human bladder cancer cell line (5637 and T24) treatment-responses to cisplatin, doxorubicin, and gemcitabine in vitro. Results: Results demonstrated consistent treatment responses between Oncopig and human cells in most concentrations tested (p>0.05). Overall, Oncopig cells were more predictive of T24 than 5637 cell therapeutic responses. Microarray analysis also demonstrated similar alterations in expression of apoptotic (GADD45B and TP53INP1) and cytoskeleton-related genes (ZMYM6 and RND1) following gemcitabine exposure between 5637 (human) and Oncopig BCCL cells, indicating apoptosis may be triggered through similar signaling pathways. Molecular docking results indicated that swine and humans had similar Dg values between the chemotherapeutics and their target proteins. Discussion: Taken together, these results suggest the Oncopig could be an attractive animal to model urothelial carcinoma due to similarities in in vitro therapeutic responses compared to human cells.

Transcriptional Profiling of Porcine HCC Xenografts Provides Insights Into Tumor Cell Microenvironment Signaling.

Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related death worldwide, representing the most common form of liver cancer. As HCC incidence and mortality continue to increase, there is a growing need for improved translational animal models to bridge the gap between basic HCC research and clinical practice to improve early detection and treatment strategies for this deadly disease. Recently the Oncopig cancer model-a novel transgenic swine model that recapitulates human cancer through Cre recombinase induced expression of KRAS G12D and TP53 R167H driver mutations-has been validated as a large animal translational model for human HCC. Due to the similar size, anatomy, physiology, immunology, genetics, and epigenetics between pigs and humans, the Oncopig has the potential to improve translation of novel diagnostic and therapeutic modalities into clinical practice. Recent studies have demonstrated the importance of tumor cells in shaping its surrounding microenvironment into one that is more proliferative, invasive, and metastatic; however, little is known about the impact of microenvironment signaling on HCC tumor biology and differential gene expression between HCC tumors and its tumor microenvironment (TME). In this study, transcriptional profiling was performed on Oncopig HCC xenograft tumors (n = 3) produced via subcutaneous injection of Oncopig HCC cells into severe combined immunodeficiency (SCID) mice. To differentiate between gene expression in the tumor and surrounding tumor microenvironment, RNA-seq reads originating from porcine (HCC tumor) and murine (microenvironment) cells were bioinformatically separated using Xenome. Principle component analysis (PCA) demonstrated clustering by group based on the expression of orthologous genes. Genes contributing to each principal component were extracted and subjected to functional analysis to identify alterations in pathway signaling between HCC cells and the microenvironment. Altered expression of genes associated with hepatic fibrosis deposition, immune response, and neo angiogenesis were observed. The results of this study provide insights into the interplay between HCC and microenvironment signaling in vivo, improving our understanding of the interplay between HCC tumor cells, the surrounding tumor microenvironment, and the impact on HCC development and progression.

Transjugular Intrahepatic Portosystemic Shunt Creation for Treatment of Gastric Varices: Systematic Literature Review and Meta-Analysis of Clinical Outcomes.

PURPOSE: To quantify the pooled clinical outcomes of stent-graft transjugular intrahepatic portosystemic shunt (TIPS) creation for the management of gastric varices (GVs) through systematic review of the literature and meta-analysis. MATERIALS AND METHODS: A PubMed and Embase search was performed from 2003 to 2020. Search terms included: (transjugular intrahepatic portosystemic shunt OR TIPS) AND (gastric varices OR fundal varices OR gastroesophageal varices OR gastroesophageal varices) AND (hemorrhage OR rebleeding OR rebleeding OR survival). Inclusion criteria spanned: English language studies, publication in peer reviewed journals, sample size ≥ 10, reported clinical outcome data, exclusive treatment of GVs (no esophageal varices), exclusive use of stent-grafts for TIPS, no chemical obliteration of GVs. Outcomes included GV rebleeding rate, overall rebleeding rate, GV occlusion rate, hepatic encephalopathy (HE) incidence, and adverse event (AE) rate. RESULTS: Literature search yielded 936 articles. Of these, 5 (0.5%) retrospective observational cohort studies met inclusion criteria, spanning 209 patients (quinquagenarian men with viral or alcoholic liver disease) with GVs treated using TIPS with adjunctive coil embolization (47%). Clinical follow-up time ranged from 4.3 to 30.6 months. Outcomes included a pooled GV rebleeding rate of 15% (95% CI: 11%, 20%), total rebleeding rate of 21% (95% CI: 15%, 27%), GV occlusion rate of 33% (95% CI: 22%, 45%), HE incidence of 30% (95% CI: 24%, 36%), and AE incidence of 3% (95% CI: 1%, 8%). CONCLUSION: The incidence of GV rebleeding after stent-graft TIPS is high. The results suggest the need for additional measures to reduce recurrent hemorrhage incidence from GVs.

Generation of genetically tailored porcine liver cancer cells by CRISPR/Cas9 editing.

Pigs provide a valuable large animal model for several diseases due to their similarity with humans in anatomy, physiology, genetics and drug metabolism. We recently generated a porcine model for TP53R167H and KRASG12D driven hepatocellular carcinoma (HCC) by autologous liver implantation. Here we describe a streamlined approach for developing genetically tailored porcine HCC cells by CRISPR/Cas9 gene editing and isolation of homogenous genetically validated cell clones. The combination of CRISPR/Cas9 editing of HCC cells described herein with the orthotopic HCC model enables development of various porcine HCC models, each with a specific mutational profile. This allows modeling the effect of different driver mutation combinations on tumor progression and in vivo testing of novel targeted therapeutic approaches in a clinically relevant large animal model.