Séminaire driving innovation Thermofisher
Lieu : IPC2
- Leveraging animal models for cancer research
- Advancing Immuno-Oncology Research and Development
- Interrogation of Three-dimensional Cell Models in Cancer Research
- Insights into Cancer Signalling
Leveraging Animal Models for Cancer Research
With the growing adoption of personalised medicine, recent advances in large scale and high throughput cellular analyses are helping streamline tools that make it easier to attain markers of disease status, examine tumour micro-environments at the single cell level and make for broad biomarker discovery and validation. Researchers today are seeking answers to questions regarding specific attributes of a tumour type, specific tumour cells and tumour micro-environment that are predictive of response in order to explore the immune system and identify pharmacodynamic biomarkers. This seminar will focus on addressing the common challenges cancer researchers face when working in animal models, and discuss recent advances and future prospects.
Advancing Immuno-Oncology Research and Development
The continued growth in immuno-oncology development and cell therapies creates a need for more robust cell-based assays. In this presentation, we will cover the topics of therapeutic antibodies, immune checkpoints, tumour microenvironments, and adoptive cell therapies including chimeric antigen receptor (CAR) T. We will review how to apply imaging, flow cytometry, and reagents to the development of each of these areas. Learn about specific applications, streamlined workflow and simple protocols performed for high-throughput data acquisition to generate deeper insight into cancer research topics.
Interrogation of Three-dimensional Cell Models in Cancer Research
Traditionally, attempts to characterise aberrant cellular processes underlying cancer progression and the development of targeted therapies have relied upon two-dimensional (2D) cell models. While useful, these models do not accurately recapitulate the complex environment of cells in the body, both normal and transformed. Increased interest in adopting three-dimensional (3D) cell models which better reflect the ‘natural’ setting has stemmed from studies demonstrating improved predictive clinical value of therapeutics that are screened in 3D versus 2D cultures. While a wide range of routine analytical methods can be performed on 3D cultures, ranging from genomic through proteomic to cell-based, fluorescence imaging in particular has lent itself to the study of these model types. When combined with antibody-based and functional biosensors, fluorescence microscopy is able to provide readouts of biological processes and regional differences throughout the sample, such as tumour spheroids, without disruption. This seminar will focus on the advantages and challenges of both labelling and imaging of structural and functional landscapes in 3D models relevant to solid tumour cell biology.
Insights into Cancer Signalling
Quantitative analysis of two cancer signalling pathways using multiplex-immunoprecipitation, targeted mass spectrometry and western blotting. The AKT/mTOR and RAS/ERK pathways represent key mechanisms for cells to regulate cell survival, proliferation, and motility. These two pathways extensively engage in cross-talk to positively and negatively regulate each other. The lack of rigorously verified reagents and a reliance on semi-quantitative immunoassays limit the accurate quantitative analysis of these pathway proteins. Immunoprecipitation coupled with mass spectrometry (IP-MS) enables the enrichment of signalling pathway proteins, revealing protein-protein interactions and PTMs and can be used for the identification of low-abundant pathway targets and for the assessment of antibody specificity. Multiplex IP coupled with MS (mIP-MS) further enhances this workflow by simultaneously quantifying multiple proteins and their phosphorylation states in a specific signalling pathway. Multiplexed IP with magnetic beads followed by targeted MS (mIP-tMS) can quantitate multiple proteins of interest, PTMs, and interacting partners in a single MS run. mIP-tMS assays were developed and optimized for absolute quantitation of targets in these pathways and benchmarked with western blot (WB). mIP-tMS assays allowed absolute quantitation of multiple total and phosphorylated targets from both pathways in low to sub-nanogram concentrations across two unstimulated, IGF-1 stimulated, and LY294002 treated cell lysates.