RESUMEN
In recent years, chimeric antigen receptor T (CAR T)-cell therapy has shown great potential in treating haematologic disease, but no breakthrough has been achieved in solid tumours. In order to clarify the antitumour mechanism of CAR T cell in solid tumours, the pharmacokinetic (PK) and pharmacodynamic (PD) investigations of CD19 CAR T cell were performed in human leukaemic xenograft mouse models. For PK investigation, we radiolabelled CD19 CAR T cell with 89 Zr and used PET imaging in the CD19-positive and the CD19-negative K562-luc animal models. For PD evaluation, optical imaging, tumour volume measurement and DNA copy-number detection were performed. Unfortunately, the qPCR results of the DNA copy number in the blood were below the detection limit. The tumour-specific uptake was higher in the CD19-positive model than in the CD19-negative model, and this was consistent with the PD results. The preliminary PK and PD studies of CD19 CAR T cell in solid tumours are instructive. Considering the less efficiency of CAR T-cell therapy of solid tumours with the limited number of CAR T cells entering the interior of solid tumours, this study is suggestive for the subsequent CAR T-cell design and evaluation of solid tumour therapy.
Asunto(s)
Antígenos CD19/inmunología , Inmunoterapia Adoptiva/métodos , Leucemia Experimental/terapia , Receptores de Antígenos de Linfocitos T/inmunología , Animales , Femenino , Humanos , Células K562 , Leucemia Experimental/diagnóstico por imagen , Ratones , Ratones Endogámicos NOD , Imagen Multimodal/métodos , Imagen Óptica/métodos , Tomografía de Emisión de Positrones/métodos , Radiofármacos/química , Radiofármacos/farmacocinética , Circonio/químicaRESUMEN
Chimeric antigen receptor (CAR) T cell therapy for hematologic malignancies is fraught with several unknowns, including number of functional T cells that engage target tumor, durability and subsequent expansion and contraction of that engagement, and whether toxicity can be managed. Non-invasive, serial imaging of CAR T cell therapy using a reporter transgene can address those issues quantitatively. We have transduced anti-CD19 CAR T cells with the prostate-specific membrane antigen (PSMA) because it is a human protein with restricted normal tissue expression and has an expanding array of positron emission tomography (PET) and therapeutic radioligands. We demonstrate that CD19-tPSMA(N9del) CAR T cells can be tracked with [18F]DCFPyL PET in a Nalm6 model of acute lymphoblastic leukemia. Divergence between the number of CD19-tPSMA(N9del) CAR T cells in peripheral blood and bone marrow and those in tumor was evident. These findings underscore the need for non-invasive repeatable monitoring of CAR T cell disposition clinically.
Asunto(s)
Antígenos de Superficie/metabolismo , Glutamato Carboxipeptidasa II/metabolismo , Inmunoterapia Adoptiva , Tomografía de Emisión de Positrones/métodos , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico por imagen , Animales , Antígenos CD19/metabolismo , Antígenos de Superficie/genética , Glutamato Carboxipeptidasa II/genética , Humanos , Leucemia Experimental/diagnóstico por imagen , Leucemia Experimental/patología , Lisina/análogos & derivados , Ratones Endogámicos NOD , Receptores de Antígenos de Linfocitos T/metabolismo , Linfocitos T/fisiología , Urea/análogos & derivadosRESUMEN
G2A is a lymphocyte-expressed G protein-coupled receptor whose genetic ablation results in the development of autoimmunity. Using HSV-TK reporter gene directed positron emission tomography (PET), we demonstrate that prior to any indication of the onset of illness, mice transplanted with BCR-ABL transduced G2A-deficient bone marrow harbor expanded populations of leukemic cells compared to recipients of wild-type bone marrow. The target cell type and anatomical locations of leukemia development are indistinguishable in animals transplanted with G2A+/+ or G2A-/- cells. Shorter disease latency in the G2A-deficient background is associated with an increased rate of cellular expansion. PET can be successfully applied to the temporal and spatial analysis of Bcr-Abl driven leukemic progression and should have utility for the study of other leukemias and lymphomas.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Proteínas de Fusión bcr-abl/fisiología , Leucemia Experimental/diagnóstico por imagen , Linfoma/diagnóstico por imagen , Proteínas Oncogénicas/genética , Receptores Acoplados a Proteínas G , Animales , Médula Ósea/patología , Proteínas de Ciclo Celular/genética , Transformación Celular Neoplásica , Cartilla de ADN/química , Herpesvirus Humano 1 , Humanos , Leucemia Experimental/genética , Leucemia Experimental/metabolismo , Linfoma/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , ARN/metabolismo , Retroviridae/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sensibilidad y Especificidad , Simplexvirus/enzimología , Simplexvirus/genética , Timidina Quinasa/genética , Timidina Quinasa/metabolismo , Tomografía Computarizada de EmisiónRESUMEN
The binding of the syngeneic monoclonal antibodies IC5F5 and 4D2B4 to Rauscher virus-induced myeloid leukemic (RMB-1) cells was analyzed in vivo in tumor-bearing BALB/c mice. To verify it these antibodies bind specifically to RMB-1 cells, purified antibodies were iodinated with the isotopes 125I and 131I. Mice previously inoculated with tumor cells were injected with these labeled monoclonal antibodies and the plasma clearance and the tissue distribution were determined. The clearance in tumor-bearing animals was faster than in control mice. The tissue distribution was corrected for nonspecific accumulation by scoring for an unrelated antibody. Calculation of a localization index showed that IC5F5 binds at least 4.5 times more specifically to tumor cells than to other tissues. A preferential localization of radioactivity in s.c. tumor tissue was seen in the scanning of animals injected with 131I-labeled antibodies. The most direct proof of specific binding was observed in autoradiograms of animals treated with 125I-labeled antibodies. Small islands of tumor cells in the livers of mice inoculated i.v. had a high density of grains compared to other tissues and also compared to tumor cells in mice treated with unrelated monoclonal antibodies. These results show efficient targeting of these monoclonal antibodies and make immunotherapy of these myeloid leukemic cells possible.
Asunto(s)
Anticuerpos Monoclonales , Radioisótopos de Yodo , Leucemia Experimental/diagnóstico por imagen , Leucemia Mieloide/diagnóstico por imagen , Animales , Anticuerpos Monoclonales/análisis , Anticuerpos Monoclonales/genética , Especificidad de Anticuerpos , Autorradiografía , Femenino , Isoanticuerpos/análisis , Isoanticuerpos/genética , Leucemia Experimental/metabolismo , Leucemia Experimental/patología , Leucemia Mieloide/metabolismo , Leucemia Mieloide/patología , Tasa de Depuración Metabólica , Ratones , Ratones Endogámicos BALB C , Cintigrafía , Virus Rauscher , Distribución TisularRESUMEN
Chelate-derivatized monoclonal antibody labeled with paramagnetic gadolinium-3+ ion has been evaluated as a tumor-specific contrast-enhancing agent in nuclear magnetic resonance imaging in the Rauscher murine erythroleukemia system. With 10(-7) M concentrations of Gd3+ delivered to the tumor target, a small but reproducible difference in proton relaxation times (T1S) was observed in excised tumors. Nuclear magnetic resonance imaging of animals, however, failed to show significant contrast enhancement of the tumor; by comparison, gamma camera images with 153Gd-labeled specific antibody did permit clear tumor visualization without subtraction. The potential use of monoclonal antibodies in tumor imaging appears to be far greater in gamma camera and positron imaging than in nuclear magnetic resonance imaging.
Asunto(s)
Anticuerpos Monoclonales , Gadolinio , Leucemia Experimental/diagnóstico por imagen , Espectroscopía de Resonancia Magnética , Radioisótopos , Animales , Anticuerpos Monoclonales/metabolismo , Femenino , Gadolinio/metabolismo , Aumento de la Imagen , Leucemia Eritroblástica Aguda/diagnóstico por imagen , Leucemia Experimental/metabolismo , Ratones , Ratones Endogámicos BALB C , Radioisótopos/metabolismo , Cintigrafía , Virus Rauscher , Neoplasias del Bazo/diagnóstico por imagenRESUMEN
High-resolution gamma camera images of mouse erythroid tumors were obtained by use of leukemia cell-specific monoclonal antibodies labeled with bifunctional radioactive metal chelates. Small tumors (200 to 300 milligrams) were visible without subtraction or enhancement 1 to 5 hours after injection of antibody. Chelate-derivitized monoclonal antibodies permit targeting of a broad spectrum of radioisotopes, including those that are optimum for agamma for gamma camera imaging or positron tomography, as well as those that are tumoricidal.