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Updated: Aug 6, 2026
A Method for Lineage Tracing of Corneal Cells Using Multi-color Fluorescent Reporter Mice
Published on: December 18, 2015
The development of single-cell lineage tracing technology and its application in immunotherapy
Jun Wang1, Zhisen Li1, Jia Chen1
1School of Life Sciences and Bio-pharmaceutics, the First Affiliated Hospital, Guangdong Pharmaceutical University, Guangzhou, People's Republic of China.
Abstract:
Immunotherapy stands as one of the most promising approaches in cancer treatment, with engineered T cell therapies, particularly chimeric antigen receptor T cell (CAR-T), leading the charge. However, relapse in some patients post-treatment suggests that research in this field remains incomplete. Tumor heterogeneity and the complexities of the immune microenvironment hinder a comprehensive understanding of the changes engineered T cells undergo once introduced into the human body. Single-cell lineage tracing (SCLT) technology facilitates the investigation of cellular development by monitoring the fate and differentiation of individual cells and their descendants within an organism. Employing methodologies such as CRISPR-based labeling and mitochondrial DNA tracking, SCLT allows for dynamic analysis of T cell clonal evolution, exhaustion mechanisms, and memory cell generation. This approach offers single-cell resolution data that contribute to resolving pertinent clinical challenges. This article provides a comprehensive review of recent developments and characteristics of the SCLT multi-omics approach. It elucidates the manner in which SCLT addresses the conventional constraints associated with spatiotemporal resolution and introduces a novel methodology for generating DNA barcodes to monitor CAR-T cells via CRISPR technology. These contributions offer valuable perspectives for the enhancement of cell therapy strategies.
Insights
Single-cell lineage tracing (SCLT) offers new ways to track engineered T cells in cancer therapy. This technology helps understand T cell behavior and improve treatments, addressing challenges like tumor heterogeneity and relapse.
Area of Science:
- Immunology
- Genetics
- Biotechnology
Background:
- Immunotherapy, especially CAR-T cell therapy, shows promise for cancer treatment.
- Patient relapse after CAR-T therapy highlights the need for deeper understanding of T cell dynamics.
- Tumor heterogeneity and the immune microenvironment complicate the study of engineered T cells in vivo.
Purpose of the Study:
- To review recent advancements in single-cell lineage tracing (SCLT) multi-omics.
- To explore how SCLT overcomes limitations in spatiotemporal resolution for cell tracking.
- To introduce a novel CRISPR-based DNA barcoding method for monitoring CAR-T cells.
Main Methods:
- Utilizes CRISPR-based labeling and mitochondrial DNA tracking for SCLT.
- Employs multi-omics approaches for comprehensive cellular analysis.
- Develops novel DNA barcodes for tracking CAR-T cell evolution.
Main Results:
- SCLT provides single-cell resolution data on T cell clonal evolution and exhaustion.
- The reviewed SCLT methods address spatiotemporal resolution constraints in cell tracking.
- A new CRISPR-based method for monitoring CAR-T cells has been developed.
Conclusions:
- SCLT is a powerful tool for understanding engineered T cell behavior in cancer therapy.
- This technology offers insights into T cell dynamics, aiding in overcoming treatment resistance.
- SCLT advancements, including novel barcoding, are crucial for enhancing future cell therapy strategies.
