Perspectives on PCR-Based Cellular Kinetics and Biodistribution Analysis for Emerging Adoptive Cell Therapies

Abstract A growing number of chimeric antigen receptor (CAR) T-cell therapies have been developed and investigated in clinical studies, with several FDA-approved therapies targeting CD19 and BCMA antigens on hematological malignancies. To further expand the indication to solid tumors and autoimmune diseases, a new generation of CAR T-cell therapies has been investigated to explore new tumor-associated antigens, allogeneic options, and cytokine armoring strategies, etc. to overcome the current limitations, including safety concerns, relapse rates, tumor microenvironment challenges, patient accessibility and manufacturing complexities. Cellular kinetics and biodistribution assessments are crucial in understanding the efficacy and safety of cell therapy, as a living drug that typically exhibits four distinct phases: distribution, expansion, contraction, and persistence within the body. Droplet digital PCR has emerged as the technology of choice in cell and gene therapy for transgene quantification with higher sensitivity, specificity, reproducibility, and absolute quantification, and as an end-point PCR provides higher tolerance to PCR inhibition. In the meantime, several unique challenges remain to be addressed in the cellular kinetics and biodistribution studies of CAR T, depending on the program stage, types of immune cells, and target indication (hematological and solid tumors, autoimmune). This opinion paper discusses the challenges and considerations of PCR-based cellular kinetics and biodistribution assessment to support emerging adoptive cell therapy programs. Graphical Abstract