A 31-plex panel for high-dimensional single-cell analysis of murine preclinical models of solid tumors by imaging mass cytometry

Abstract

Currently, the study of resistance mechanisms and disease progression in cancer relies on the capacity to analyze tumors as a complex ecosystem of healthy and malignant cells. Therefore, one of the current challenges is to decipher the intra-tumor heterogeneity and especially the spatial distribution and interactions of the different cellular actors within the tumor. Preclinical mouse models are widely used to extend our understanding of the tumor microenvironment (TME). Such models are becoming more sophisticated and allow investigating questions that cannot be addressed in clinical studies. Indeed, besides studying the tumor cell interactions within their environment, mouse models allow evaluating the efficacy of new drugs and delivery approaches, treatment posology, and toxicity. Spatially resolved analyses of the intra-tumor heterogeneity require global approaches to identify and localize a large number of different cell types. For this purpose, imaging mass cytometry (IMC) is a major asset in the field of human immuno-oncology. However, the paucity of validated IMC panels to study TME in pre-clinical mouse models remains a critical obstacle to translational or basic research in oncology. Here, we validated a panel of 31 markers for studying at the single-cell level the TME and the immune landscape for discovering/characterizing cells with complex phenotypes and the interactions shaping the tumor ecosystem in mouse models.

Multiplexed Imaging Mass Cytometry Analysis in Preclinical Models of Pancreatic Cancer

“…For these reasons, IMC has been widely used in many studies aimed at dissecting the composition of different types of human cancer [21,22]. By contrast, only a few studies report the use of IMC in preclinical models of diseases [23][24][25][26][27].…”

“…These studies are particularly interesting since they facilitate the development of IMC applications in the oncology field by providing panels of antibodies usable for IMC analysis [44][45][46][47]. However, this technique has been mostly applied to human samples and only a few studies reported the use of IMC technology on mouse models of carcinogenesis [23,26]. To our knowledge, this is the first study reporting the validation of an IMC antibody panel for the investigation of a mouse model of pancreatic cancer.…”

Spatial mapping of rheumatoid arthritis synovial niches reveals specific macrophage networks associated with response to therapy

“…Cell segmentation was performed as previously described ( 49 ). Briefly, pixel classification and training were performed using Ilastik v1.3.3 to generate a probability map.…”

Integrated spatial and multimodal single‐cell transcriptomics reveal patient‐dependent cell heterogeneity in splenic marginal zone lymphoma

“…The principle of the assay has been described previously [28]. Tissue labeling and data acquisition using the Hyperion Imaging System (Standard Biotools, San Francisco, CA, USA) were performed following the manufacturer's instructions, as described previously [29]. Further details are provided in Supplementary materials and methods.…”