In vitro characterization of oncolytic virus in combination with STAT3 inhibition for improved therapy against pancreatic ductal adenocarcinoma

Table of Contents

I. Introduction

1.1 Pancreatic cancer.

1.1.1 PDAC development and progression

1.1.2 Microenvironment

1.1.3 Current treatment options and their limitations

1.2 Oncolytic viruses

1.2.1 Vesicular stomatitis virus

1.3 Signal transducer and activator of transcription 3

1.4 Aim of this thesis

II. Material

2.1 Cell lines

2.2 Reagents

2.3 Antibodies

2.4 Consumables

2.5 Appliances

III. Methods

3.1 Cell culture

3.1.1 3D cell culture systems

3.1.2 Co-culture systems

3.2 Viability assay

3.3 Tissue culture infection dose

3.4 Western blotting

3.5 Flow cytometric analysis

3.6 Confocal microscopy

3.7 Luciferase assay

3.7.1 Plasmid DNA preparation

3.7.2 Transfection

3.7.3 Interferon induction and response

3.8 Scratch assay

IV. Results

4.1 PDAC cells are susceptible to rVSV-GFP-mediated oncolysis in vitro

4.2 S3I-201 causes reduction of PDAC cell viability in vitro

4.3 Combination therapy enhances PDAC killing and safety

4.4 Combinational therapy in the microenvironment

4.5 Effects of S3I-201 and VSV in co-culture

4.6 Combination therapy in 3D

4.7 Inhibition of migration

V. Discussion

Research Objectives and Topics

This master thesis investigates the therapeutic potential of combining oncolytic vesicular stomatitis virus (VSV) with STAT3 inhibition as a novel treatment strategy for pancreatic ductal adenocarcinoma (PDAC). The study focuses on the synergistic effects of these agents on tumor cell killing, the reduction of migratory and proliferative potential in cancer-associated fibroblasts, and the potential to enhance therapeutic safety in healthy primary pancreatic cells.

• Oncolytic virotherapy with rVSV-GFP
• STAT3 signaling inhibition using S3I-201
• PDAC microenvironment and tumor-stroma crosstalk
• 2D, 3D, and co-culture model development
• Mechanisms of tumor cell death and migration inhibition

Excerpt from the Book

Summary of Chapters

I. Introduction: Provides an overview of pancreatic ductal adenocarcinoma, the role of oncolytic viruses, the function of STAT3 signaling, and the specific goals of this thesis.

II. Material: Lists the cell lines, reagents, antibodies, consumables, and laboratory equipment used throughout the experiments.

III. Methods: Details the experimental procedures, including cell culture techniques (2D, 3D, and co-culture), viability assays, viral titer determination, western blotting, and microscopy methods.

IV. Results: Presents the findings regarding the synergistic effects of rVSV-GFP and STAT3 inhibitor S3I-201 in various PDAC models, including infection efficacy, cell viability, and migration potential.

V. Discussion: Evaluates the experimental results, interprets the synergy between the two therapeutic agents, discusses implications for future clinical applications, and addresses limitations and future research directions.

Keywords

Pancreatic ductal adenocarcinoma, PDAC, oncolytic virus, vesicular stomatitis virus, VSV, STAT3, S3I-201, cancer-associated fibroblasts, CAFs, tumor microenvironment, synergistic therapy, apoptosis, cell migration, tumor spheroids, immunotherapy.

Frequently Asked Questions

What is the core focus of this research?

The research explores the potential of a combinational therapy using oncolytic viruses and STAT3 inhibitors to improve the treatment of pancreatic ductal adenocarcinoma.

What are the primary themes covered in this thesis?

The thesis covers tumor biology, viral oncology, signaling pathways like STAT3, the tumor microenvironment, and advanced in vitro modeling.

What is the main research question or goal?

The goal is to determine if the combination of rVSV-GFP and the STAT3 inhibitor S3I-201 exhibits synergistic anti-tumor effects in PDAC cells while maintaining or improving safety for healthy cells.

Which scientific methods are employed?

The study utilizes 2D cell cultures, 3D tumor spheroids, transwell co-culture systems, MTT viability assays, flow cytometry, western blotting, and confocal microscopy.

What is addressed in the main body of the work?

The main body details the materials and methods used for cell culture and assays, followed by extensive results on cell viability, viral replication, signaling pathway inhibition, and migration studies.

Which keywords define this work?

Key terms include PDAC, oncolytic virus, STAT3 inhibition, tumor microenvironment, and combinatorial therapy.

How does STAT3 inhibition impact the virus-PDAC interaction?

The study indicates that STAT3 inhibition by S3I-201 enhances the cytotoxic effect against PDAC cells without interfering with viral replication in tumor cells.

What was observed regarding healthy primary pancreatic cells?

The combination therapy was found to be significantly safer for healthy primary cells, as the inhibitor reduced viral titers in healthy tissue, suggesting a potential improvement in therapeutic safety.