An End-to-End Workflow for Interrogating Tumor-Infiltrating Myeloid Cells Using Mass Cytometry

Area of Science:

• Immunology
• Cancer Biology
• Biotechnology

Background:

• Myeloid cells are key immune components with diverse roles in tumor progression.
• Specific myeloid subsets, like tumor-associated macrophages and myeloid-derived suppressive cells (MDSCs), promote tumor growth.
• Understanding myeloid cell heterogeneity is crucial for developing effective cancer immunotherapies.

Purpose of the Study:

• To provide a comprehensive end-to-end workflow for deep phenotyping of tumor-infiltrating myeloid cells.
• To enable detailed characterization of myeloid cell heterogeneity using mass cytometry.
• To facilitate the interrogation of phosphoproteins in myeloid cells within the tumor microenvironment.

Main Methods:

• Utilized mass cytometry for high-dimensional single-cell analysis of myeloid cells.
• Developed and validated an experimental protocol for sample preparation and staining.
• Implemented computational analysis including Phenograph clustering for myeloid cell subset identification.

Main Results:

• Established a robust workflow for deep phenotyping of tumor-infiltrating myeloid cells.
• Enabled comprehensive characterization of myeloid cell heterogeneity and functional states.
• Provided protocols for phosphoprotein analysis in circulating and tumor-infiltrating myeloid cells.

Conclusions:

• Mass cytometry offers an unprecedented opportunity to dissect myeloid cell heterogeneity in tumors.
• The provided workflow and computational scripts facilitate advanced myeloid cell profiling.
• This research supports the development of targeted cancer immunotherapies by elucidating myeloid cell functions.