A global live cell barcoding approach for multiplexed mass cytometry profiling of mouse tumors

Area of Science:

• Cancer immunology
• Mass cytometry
• Tumor microenvironment (TME) analysis

Background:

• Mass cytometry is crucial for cancer immunology research, particularly for analyzing the tumor microenvironment (TME).
• Multiplexing samples using barcoding enhances data acquisition and analysis efficiency.
• A reliable method for barcoding mouse cancer cells, alongside immune cells, was previously lacking.

Purpose of the Study:

• To develop and validate a novel barcoding strategy for mouse cancer cells in mass cytometry.
• To enable simultaneous barcoding of both tumor and immune cells for comprehensive TME analysis.
• To apply this method to study tumor-immune interactions in different mouse cancer models.

Main Methods:

• Identification of CD29 and CD29 as widely expressed markers on mouse cancer cell lines.
• Conjugation of anti-CD29 and anti-CD98 antibodies to metal isotopes for barcoding.
• Validation of a 10-plex barcoding system using CD29, CD98, and CD45 (for immune cells).
• Application to multiplexing 10 tumors from MC38 and KPC models.

Main Results:

• CD29 and CD98 were validated as effective markers for barcoding mouse cancer cells.
• A 10-plex barcoding system combining tumor and immune cell markers was successfully established.
• The method allowed for the recapitulation of known differences in the PD1-PDL1 axis between MC38 and KPC models.
• Simultaneous interrogation of tumor cells and immune cells within the TME was achieved.

Conclusions:

• The developed barcoding system enables efficient multiplexing of mouse tumors and immune cells.
• This method facilitates deeper investigation of tumor-immune interactions in the TME.
• The approach holds significant potential for advancing cancer immunology research and therapeutic response characterization.