Development of a Clinically Relevant Reporter for Chimeric Antigen Receptor T-cell Expansion, Trafficking, and Toxicity

Abstract

Although chimeric antigen receptor T (CART)-cell therapy has been successful in treating certain hematologic malignancies, wider adoption of CART-cell therapy is limited because of minimal activity in solid tumors and development of life-threatening toxicities, including cytokine release syndrome (CRS). There is a lack of a robust, clinically relevant imaging platform to monitor <i>in vivo</i> expansion and trafficking to tumor sites. To address this, we utilized the sodium iodide symporter (NIS) as a platform to image and track CART cells. We engineered CD19-directed and B-cell maturation antigen (BCMA)-directed CART cells to express NIS (NIS⁺CART19 and NIS⁺BCMA-CART, respectively) and tested the sensitivity of ¹⁸F-TFB-PET to detect trafficking and expansion in systemic and localized tumor models and in a CART-cell toxicity model. NIS⁺CART19 and NIS⁺BCMA-CART cells were generated through dual transduction with two vectors and demonstrated exclusive ¹²⁵I uptake <i>in vitro</i>. ¹⁸F-TFB-PET detected NIS⁺CART cells <i>in vivo</i> to a sensitivity level of 40,000 cells. ¹⁸F-TFB-PET confirmed NIS⁺BCMA-CART-cell trafficking to the tumor sites in localized and systemic tumor models. In a xenograft model for CART-cell toxicity, ¹⁸F-TFB-PET revealed significant systemic uptake, correlating with CART-cell <i>in vivo</i> expansion, cytokine production, and development of CRS-associated clinical symptoms. NIS provides a sensitive, clinically applicable platform for CART-cell imaging with PET scan. ¹⁸F-TFB-PET detected CART-cell trafficking to tumor sites and <i>in vivo</i> expansion, correlating with the development of clinical and laboratory markers of CRS. These studies demonstrate a noninvasive, clinically relevant method to assess CART-cell functions <i>in vivo</i>.