scholarly journals Characterisation of tumour microenvironment remodelling following oncogene inhibition in preclinical studies with imaging mass cytometry

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Febe van Maldegem ◽  
Karishma Valand ◽  
Megan Cole ◽  
Harshil Patel ◽  
Mihaela Angelova ◽  
...  
Keyword(s):  
Tumour Microenvironment ◽  
Mass Cytometry ◽  
Effector Cells ◽  
Genetic Manipulations ◽  
Mouse Tissues ◽  
Multiplexed Imaging ◽  
Lung Cancer Model ◽  
Antigen Presenting ◽  
Source Of Information ◽  
Immune Competent Mouse

AbstractMouse models are critical in pre-clinical studies of cancer therapy, allowing dissection of mechanisms through chemical and genetic manipulations that are not feasible in the clinical setting. In studies of the tumour microenvironment (TME), multiplexed imaging methods can provide a rich source of information. However, the application of such technologies in mouse tissues is still in its infancy. Here we present a workflow for studying the TME using imaging mass cytometry with a panel of 27 antibodies on frozen mouse tissues. We optimise and validate image segmentation strategies and automate the process in a Nextflow-based pipeline (imcyto) that is scalable and portable, allowing for parallelised segmentation of large multi-image datasets. With these methods we interrogate the remodelling of the TME induced by a KRAS G12C inhibitor in an immune competent mouse orthotopic lung cancer model, highlighting the infiltration and activation of antigen presenting cells and effector cells.

scholarly journals Characterisation of tumour immune microenvironment remodelling following oncogene inhibition in preclinical studies using an optimised imaging mass cytometry workflow

2021 ◽  
Author(s):  
Febe van Maldegem ◽  
Karishma Valand ◽  
Megan Cole ◽  
Harshil Patel ◽  
Mihaela Angelova ◽  
...  
Keyword(s):  
Tumour Microenvironment ◽  
Cancer Model ◽  
Mass Cytometry ◽  
Genetic Manipulations ◽  
Mouse Tissues ◽  
Wide Range ◽  
Multiplexed Imaging ◽  
Lung Cancer Model ◽  
Source Of Information ◽  
Immune Competent Mouse

AbstractMouse models are critical in pre-clinical studies of cancer therapy, allowing dissection of mechanisms through chemical and genetic manipulations that are not feasible in the clinical setting. In studies of the tumour microenvironment (TME), novel highly multiplexed imaging methods can provide a rich source of information. However, the application of such technologies in mouse tissues is still in its infancy. Here we present a workflow for studying the TME using imaging mass cytometry with a panel of 27 antibodies on frozen mouse tissues. We optimised and validated image segmentation strategies and automated the process in a Nextflow-based pipeline (imcyto) that is scalable and portable, allowing for parallelised segmentation of large multi-image datasets. Incorporating user-specific plugins, imcyto can be flexibly tailored to a wide range of segmentation needs. With these methods we interrogated the dramatic remodelling of the TME induced by a KRAS G12C inhibitor in an immune competent mouse orthotopic lung cancer model, showcasing their potential as key discovery tools to enhance understanding of the interplay between tumour, stroma, and immune cells in the spatial context of the tissue.

scholarly journals Characterisation of tumour immune microenvironment remodelling following oncogene inhibition in preclinical studies using an optimised imaging mass cytometry workflow

2021 ◽  
Author(s):  
Febe van Maldegem ◽  
Karishma Valand ◽  
Megan Cole ◽  
Harshil Patel ◽  
Mihaela Angelova ◽  
...  
Keyword(s):  
Tumour Microenvironment ◽  
Cancer Model ◽  
Mass Cytometry ◽  
Genetic Manipulations ◽  
Mouse Tissues ◽  
Wide Range ◽  
Multiplexed Imaging ◽  
Lung Cancer Model ◽  
Source Of Information ◽  
Immune Competent Mouse

Abstract Mouse models are critical in pre-clinical studies of cancer therapy, allowing dissection of mechanisms through chemical and genetic manipulations that are not feasible in the clinical setting. In studies of the tumour microenvironment (TME), novel highly multiplexed imaging methods can provide a rich source of information. However, the application of such technologies in mouse tissues is still in its infancy. Here we present a workflow for studying the TME using imaging mass cytometry with a panel of 27 antibodies on frozen mouse tissues. We optimised and validated image segmentation strategies and automated the process in a Nextflow-based pipeline (imcyto) that is scalable and portable, allowing for parallelised segmentation of large multi-image datasets. Incorporating user-specific plugins, imcyto can be flexibly tailored to a wide range of segmentation needs. With these methods we interrogated the dramatic remodelling of the TME induced by a KRAS G12C inhibitor in an immune competent mouse orthotopic lung cancer model, showcasing their potential as key discovery tools to enhance understanding of the interplay between tumour, stroma, and immune cells in the spatial context of the tissue.

scholarly journals Understanding the tumour immune microenvironment of stage III colorectal cancer using multiplexed imaging mass cytometry

Pathology ◽  
2021 ◽  
Vol 53 ◽  
pp. S36-S37
Author(s):  
Minh Tran ◽  
Andrew Su ◽  
HoJoon Lee ◽  
Richard Cruz ◽  
Lance Pflieger ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Stage Iii ◽  
Mass Cytometry ◽  
Immune Microenvironment ◽  
Multiplexed Imaging

scholarly journals Simultaneous Multiplexed Imaging of mRNA and Proteins with Subcellular Resolution in Breast Cancer Tissue Samples by Mass Cytometry

Cell Systems ◽  
2018 ◽  
Vol 6 (4) ◽  
pp. 531 ◽  
Author(s):  
Daniel Schulz ◽  
Vito Riccardo Tomaso Zanotelli ◽  
Jana Raja Fischer ◽  
Denis Schapiro ◽  
Stefanie Engler ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Breast Cancer Tissue ◽  
Cancer Tissue ◽  
Mass Cytometry ◽  
Tissue Samples ◽  
Multiplexed Imaging ◽  
Subcellular Resolution

scholarly journals Ly6Chi Monocytes in the Inflamed Colon Give Rise to Proinflammatory Effector Cells and Migratory Antigen-Presenting Cells

Immunity ◽  
2012 ◽  
Vol 37 (6) ◽  
pp. 1076-1090 ◽  
Author(s):  
Ehud Zigmond ◽  
Chen Varol ◽  
Julia Farache ◽  
Elinor Elmaliah ◽  
Ansuman T. Satpathy ◽  
...  
Keyword(s):  
Antigen Presenting Cells ◽  
Effector Cells ◽  
Antigen Presenting

Potent Anti-Leukemic Activity of a Cationic Lipid-DNA Complex.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4891-4891
Author(s):  
Zachary Herse ◽  
Stella Chang ◽  
Denny Liggitt ◽  
Jeffery Fairman ◽  
David F. Claxton
Keyword(s):  
Cationic Lipid ◽  
Green Fluorescence Protein ◽  
Acute Leukemias ◽  
Immune Effector ◽  
Effector Cells ◽  
Bacterial Plasmid ◽  
Fluorescence Protein ◽  
Cellular Immune ◽  
Antigen Presenting ◽  
Dna Complex

Abstract Cationic Lipid and DNA Complex (CLDC) is a chemically defined preparation of lipids and non-coding bacterial plasmid DNA able to activate antigen presenting and immune effector cells and thereby enhance cellular immune responses. We tested this preparation subcutaneously (SC) in two models of transplantable murine leukemia: 32D -(bcr-abl)p210 (leukemogenic in C3H/HEJ animals) and WEHI-3B cells (leukemogenic in BALB-C mice.) In multiple experiments using the 32Dp210 model a single SC dose of CLDC (200 ul) prevented or significantly delayed death from leukemia when delivered between -1 and 16 days from leukemic challenge. Specifically, control animals died at 28 days (SD=2.73) of leukemia whereas 20/24 similarly leukemia challenged but CLDC treated animals were alive >71 days (p<0.0001 − 4/20 animals died of leukemia). Animals dying of leukemia had swollen spleens and leukemic infiltration of spleen and liver on histopathological evaluation. Similarly Balb-C animals challenged with WEHI-3B cells died at a median of 30 days unless treated with CLDC (1 dose day 6 − 4/5 alive >day 40 p=0.0017). A green fluorescence protein (GFP) expressing subclone of 32Dp210 allowed quantitation of leukemia in peripheral blood (PB). PB leukocytes were 20% GFP positive on day 19. After CLDC on day 19, day 21 PB showed only 2% GFP. Animals receiving 32D-p210-GFP and treated with CLDC and CD8 antibody on day 19 showed higher GFP in followup and died earlier (median day 35) than animals treated with CLDC alone (>42 days) but later than animals receiving no treatment after leukemic challenge (median 23 days, p=0.0002.) See Figure below. We conclude that CLDC shows potent anti-leukemia activity in two murine models of AML. Antibody experiments suggest that CD8 positive effector cells contribute to this effect. Additional experiments are maturing and will be reported. This preparation has promise for activity in human acute leukemias. Figure Figure

Mast cells are crucial for early inflammation, migration of Langerhans cells, and CTL responses following topical application of TLR7 ligand in mice

Blood ◽  
2007 ◽  
Vol 110 (3) ◽  
pp. 946-953 ◽  
Author(s):  
Valeska Heib ◽  
Marc Becker ◽  
Tobias Warger ◽  
Gerd Rechtsteiner ◽  
Christine Tertilt ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Topical Application ◽  
Langerhans Cells ◽  
Cytotoxic T Lymphocyte ◽  
Effector Cells ◽  
Tnf Α ◽  
Transcutaneous Immunization ◽  
Antigen Presenting ◽  
Tlr7 Ligand

Abstract Until recently, IgE-activated mast cells have been regarded merely as effector cells of adaptive immune responses, involved in allergic reactions and mucosal immunity to parasites. Herein, we report that murine dermal mast cells, activated by local administration of a cream containing the synthetic TLR7 ligand imiquimod, are essential to initiate an early inflammatory reaction. The mast-cell–derived cytokines TNF-α and IL-1β play an important role in this process. Furthermore, TLR7-activated mast cells are also able to promote the emigration of Langerhans cells, which partly depends on the expression of mast-cell–derived IL-1β. We have previously shown that TLR7 ligation enhances transcutaneous immunization evoked by topical application of vaccine antigens to the skin, a procedure that directly targets skin-resident antigen-presenting cells. Consequently, we now demonstrate here that the capacity to mount a peptide-specific cytotoxic T-lymphocyte response following transcutaneous immunization using imiquimod as adjuvant is severely impaired in mast-cell–deficient mice. Thus, these findings demonstrate the potent versability of alternatively activated mast cells at the interface of innate and adaptive immunity.

scholarly journals Dice-XMBD: Deep Learning-Based Cell Segmentation for Imaging Mass Cytometry

2021 ◽  
Vol 12 ◽  
Author(s):  
Xu Xiao ◽  
Ying Qiao ◽  
Yudi Jiao ◽  
Na Fu ◽  
Wenxian Yang ◽  
...  
Keyword(s):  
Deep Learning ◽  
High Resolution ◽  
Single Cell ◽  
Image Data ◽  
Basic Research ◽  
Cell Segmentation ◽  
Imaging Method ◽  
Imaging Data ◽  
Mass Cytometry ◽  
Multiplexed Imaging

Highly multiplexed imaging technology is a powerful tool to facilitate understanding the composition and interactions of cells in tumor microenvironments at subcellular resolution, which is crucial for both basic research and clinical applications. Imaging mass cytometry (IMC), a multiplex imaging method recently introduced, can measure up to 100 markers simultaneously in one tissue section by using a high-resolution laser with a mass cytometer. However, due to its high resolution and large number of channels, how to process and interpret the image data from IMC remains a key challenge to its further applications. Accurate and reliable single cell segmentation is the first and a critical step to process IMC image data. Unfortunately, existing segmentation pipelines either produce inaccurate cell segmentation results or require manual annotation, which is very time consuming. Here, we developed Dice-XMBD1, a Deep learnIng-based Cell sEgmentation algorithm for tissue multiplexed imaging data. In comparison with other state-of-the-art cell segmentation methods currently used for IMC images, Dice-XMBD generates more accurate single cell masks efficiently on IMC images produced with different nuclear, membrane, and cytoplasm markers. All codes and datasets are available at https://github.com/xmuyulab/Dice-XMBD.

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