The Spatial Biology Week™ 2022

Multiple clinical trials have shown an overall improved response rate in cancers by combining radiotherapy (RT) with immunotherapy (IO), however, treatment rates remain low and unpredictable. The suboptimal outcomes are largely due to the lack of knowledge of the underlying immunomodulation effect and an effective treatment-response biomarker. Previous studies using tissue-based assays like multiplexed immunofluorescence (mIF) have demonstrated that cellular spatial organization within the tumor microenvironment represents a critical factor influencing anti-tumor immunity. Hence, we sought to characterize the in-situ molecular immune response of RT-treated cancer tissues by using multiple spatial and immunology technologies, to advance our understanding of RT-induced immunomodulation effect and its synergistic benefits with IO.

Dissertation points:

• Desired features of spatial biology technologies
• Relationship between tissue biomarkers and liquid biopsies in oncology
• The next in the quest to understand cancer tissue microenvironment dynamics

• In the context of biomarker discovery, what are the common challenges around the analysis of tissue 
• How can spatial proteomics be used to overcome these
• What approaches does Propath UK use to streamline multiplexed IF/IHC 
• Designing biomarker discovery projects using spatial technologies

Multiplex immunofluorescence platforms capable of staining 40+ markers on a single 4 mm tissue are an incredible tool for interrogating the pathology of a variety of disease processes, in particular immune responses to cancers. While these platforms are capable of providing deep and high-resolution multimarker and spatial data from a single sample, there are specific risks to guard against. These include artifacts related to sensitivity, dynamic range, spectral bleed, and signal carryover from staining cycles. Of course, any significant observation made in multiplex IF must be corroborated using orthogonal measures, but due to the expense and effort required to perform the experiments, the potential impact of the results, and the precious nature of the samples, a properly validated assay is required. There is currently no standard validation approach for multiplex IF. Here we review our first experience with a beta unit of the Lunaphore COMET, inclusive of operation, performance, panel development, as well as a prototype validation

• COMET technology walkthrough + mIF landscape
• What a multiplex validation should cover
• Comet validation plan
• What worked & lessons learned

Discussion points: 

• Multiplex panel validation: challenges and opportunities
• Ideal technological requirements in a biopharma setting
• Spatially-resolved biomarkers use in a clinical setting

• The added value of in situ spatial proteomics
• The next era of pathology – from single plex to broad panels
• How can you access the latest spatial omics technologies for your research

The SARS-CoV-2 virus is not only associated with respiratory tract infection, but can also infect other organs such as the placenta, resulting in unique clinical syndromes depending on the organ infected. Through the use of multiomic spatial profiling of SARS-CoV-2 infected tissue, including lung and placenta, we demonstrate that:

• Detection of virus in tissue requires a robust muitiomic approach
• SARS-CoV-2 infection elicits a unique inflammatory response
• Virus infection in lung is associated with dysregulated collagen synthesis
• Placental infection is typified by competing anti-viral and immune evasive responses.

Discussion points:

• Pros and Cons of an open technology platform for spatial biology
• Spatial protein versus RNA biomarkers
• Most promising fields outside oncology where spatial biology can make a long-lasting impact

The identification of the markers expressed in the tumor microenvironment (TME) and their spatial distribution is key to understanding tumor growth and heterogeneity, as well as to direct therapeutic selection.

In this 30-min webinar, Dr. Ghislaine Lioux will present how the HALO® and HALO AI™ software solutions can be used to analyze images of immuno-oncology panels encompassing a 20-biomarker panel from the COMET™ platform.

During a live demonstration of the HALO and HALO AI software, you will learn:

• How to intuitively segment the tumoral and stromal compartments with artificial intelligence
• How to phenotype cells and map their spatial distribution, density, and interactions
• How to automate the analysis of highly variable samples and overcome artefacts in tissue microarrays

• A guided bi-directional workflow tailored for setting phenotypes with continuous QC and review of results
• Powerful pre-trained nuclear detection APPs for multiplex immunofluorescence and IMC (imaging mass cytometry).
• Easy-to-use channel management tools to QC images and review biomarker localization. Group your channels of interest in multiple meaningful 7-color channel groups to quickly toggle between panels.
• An advanced interactive Data Exploration and QC toolbox including, t-SNE, Scatter and Box-plotting capabilities

Join this talk to discover more on:

• Interactive visualization and computational analysis of high-dimensional immunofluorescence slide microscopy imaging data for cancer research and molecular cancer diagnostics
• Combination of human expert annotation, machine learning-based computer vision, and spatial statistics for quantitative single-cell analysis to describe and model patterns of immune cell infiltrates and interactions between individual cells in spatial tissue context
• Discovery and development of quantitative tissue imaging biomarkers for molecularly targeted therapy and immunotherapy