Development of an in situ CAR-T cell protocol through optical and PSMA-targeted PET imaging

N Nisi Zhang (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) J Jai Woong Seo (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) E Elise Robinson (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) A Angelie Rivera-Rodriguez (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) J James Wang (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) Y Yutong Guo (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) D Debra K. Czerwinski (Division of Oncology, Department of Medicine, Stanford Cancer Institute) H Ha Rin Kim (Division of Oncology, Department of Medicine, Stanford University School of Medicine) S Spencer K. Tumbale (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) M Marina N. Raie (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) B Basit L. Jan (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) G Gokce Engudar (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University) A Adrienne Sallets (Division of Oncology, Department of Medicine, Stanford Cancer Institute) I Il Minn (Department of Radiology, University of Texas Southwestern Medical Center) M Martin G. Pomper (Department of Radiology, University of Texas Southwestern Medical Center) R Ronald Levy (Division of Oncology, Department of Medicine, Stanford Cancer Institute) K Katherine W. Ferrara (Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University)

Abstract

In situ T cell transfection methods overcome the complexity and high costs associated with conventional chimeric antigen receptor (CAR)-T therapy. However, the in situ CAR-T cell approach operates within the patient’s complex immune environment and bypasses preinfusion ex vivo cellular quality controls, necessitating advanced imaging techniques to track immune cell migration and function. Positron emission tomography (PET) can detect biochemical processes in patients and, when combined with a radiotracer specific for the engineered cells, can monitor CAR-T cell trafficking. Herein, we develop an approach for in situ T cell generation, tracking, and functional assessment using anti-CD5-conjugated lipid nanoparticles for codelivering CD19 CAR mRNA (mCAR19) and a prostate-specific membrane antigen mRNA (mPSMA) tag. With interleukin-7 (IL-7) preconditioning and repeated administration, this approach achieves tumor-free survival in 75% of B cell lymphoma-bearing mice (similar efficacy to ex vivo approaches), and through PET imaging of 68 Ga-PSMA-617, the generation and tumor infiltration of in situ-engineered PSMA-tagged CD19 CAR-T cells is validated.

Article Details

Volume / Issue Vol. 122, Issue 24
Published June 17, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (17)

N

Nisi Zhang

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

J

Jai Woong Seo

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

E

Elise Robinson

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

A

Angelie Rivera-Rodriguez

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

J

James Wang

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

Y

Yutong Guo

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

D

Debra K. Czerwinski

Division of Oncology, Department of Medicine, Stanford Cancer Institute

H

Ha Rin Kim

Division of Oncology, Department of Medicine, Stanford University School of Medicine

S

Spencer K. Tumbale

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

M

Marina N. Raie

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

B

Basit L. Jan

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

G

Gokce Engudar

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University

A

Adrienne Sallets

Division of Oncology, Department of Medicine, Stanford Cancer Institute

I

Il Minn

Department of Radiology, University of Texas Southwestern Medical Center

M

Martin G. Pomper

Department of Radiology, University of Texas Southwestern Medical Center

R

Ronald Levy

Division of Oncology, Department of Medicine, Stanford Cancer Institute

K

Katherine W. Ferrara

Molecular Imaging Program at Stanford, Department of Radiology, School of Medicine, Stanford University