Cellular behavior analysis from live-cell imaging of TCR T cell-cancer cell interactions

Area of Science:

• Immunology
• Bioengineering
• Computational Biology

Background:

• T cell therapies like CAR T and TCR T cells are advancing cancer treatment.
• Engineering cellular behavior is crucial for expanding T cell therapy applications.
• Live-cell imaging is a common method for evaluating engineered T cells.

Purpose of the Study:

• To develop novel computational tools for analyzing T cell behavior from live-cell imaging data.
• To characterize T cell phenotypes, including population dynamics, morphology, movement, and interactions.
• To investigate the impact of specific gene knockouts (RASA2, CUL5) on T cell-cancer cell interactions and therapeutic efficacy.

Main Methods:

• Development of the Caliban pipeline for deep learning-based cell segmentation and tracking.
• Development of the Occident pipeline for cataloging T cell phenotypes in co-cultures.
• Application of spatiotemporal models to quantify T cell recruitment and proliferation.
• Analysis of TCR T cells with RASA2 and CUL5 knockouts co-cultured with tumor cells.

Main Results:

• RASA2 knockout TCR T cells exhibited longer interaction times with cancer cells, enhancing T cell activation and killing efficacy.
• CUL5 knockout TCR T cells demonstrated increased proliferation rates, leading to a greater number of T cells available for targeting cancer cells.
• The Caliban and Occident pipelines effectively enabled detailed cellular behavior analysis.

Conclusions:

• The developed Caliban and Occident pipelines provide powerful tools for T cell behavior analysis.
• Genetic modifications like RASA2 and CUL5 knockouts can significantly enhance T cell therapy effectiveness.
• This approach aids in the engineering of improved T cell therapies for cancer treatment.