# Spatiotemporal profiling reveals distinct dynamics and checkpoint regulations of CAR-T and CAR-NKT cells against solid tumors

## TL;DR

This study compares CAR-T and CAR-NKT cell therapies in solid tumors, revealing differences in their behavior and response to immune checkpoint inhibitors.

## Contribution

The study provides a preclinical comparison of CAR-T and CAR-NKT cells using spatiotemporal profiling and identifies distinct checkpoint regulation patterns.

## Key findings

- CAR-NKT cells show better tumor infiltration and persistence than CAR-T cells.
- CAR-T cells respond synergistically to TIGIT blockade, while CAR-NKT cells are more sensitive to CD96 blockade.
- The study reveals divergent therapeutic dynamics and immune checkpoint profiles between CAR-T and CAR-NKT cells.

## Abstract

Chimeric antigen receptor (CAR)-engineered T (CAR-T) cell therapies have demonstrated remarkable efficacy in hematologic malignancies; however, their clinical performance in solid tumors remains limited due to suboptimal tumor infiltration, antigen heterogeneity, and immunosuppressive tumor microenvironments (TME). Invariant natural killer T (NKT) cells have recently emerged as a promising alternative platform for CAR engineering, owing to their intrinsic tissue-homing capacity, multi-modal cytotoxicity, and ability to reshape the TME. In this study, we performed a comprehensive preclinical comparison of conventional CAR-T cells and allogeneic stem cell–derived IL-15–enhanced CAR-NKT cells in solid tumor models, integrating spatiotemporal transcriptomic profiling across multiple tissues and longitudinal time points. Our analyses revealed distinct in vivo pharmacokinetic, pharmacodynamic, and immunoregulatory profiles between the two cell therapy modalities. Compared with CAR-T cells, CAR-NKT cells demonstrated superior homing, infiltration, and localization within solid tumors, along with prolonged in vivo persistence and a unique immune checkpoint receptor expression landscape. Notably, CAR-T cells exhibited synergistic antitumor responses when combined with TIGIT blockade, whereas CAR-NKT cells showed greater sensitivity to CD96 blockade in vivo. These findings highlight the divergent therapeutic dynamics of CAR-T and CAR-NKT cells and provide mechanistic insights that inform the rational design of next-generation cell therapies and combinatorial strategies for solid tumors.