Titre :

Droplet digital PCR allows vector copy number assessment and monitoring of experimental CAR T cells in murine xenograft models or approved CD19 CAR T cell-treated patients.

Auteur(s) :

Haderbache, Rafik [Auteur]
Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) [RIGHT]
Warda, W. [Auteur]
Hervouet, E. [Auteur]
Da Rocha, M. N. [Auteur]
Trad, R. [Auteur]
Allain, V. [Auteur]
Nicod, C. [Auteur]
Thieblemeont, Catherine [Auteur]
Hopital Saint-Louis [AP-HP] [AP-HP]
Boissel, N. [Auteur]
Varlet, Pauline [Auteur]
Institut de Recherche Translationnelle sur l'Inflammation (INFINITE) - U1286
Yakoub-Agha, Ibrahim [Auteur]
Institute for Translational Research in Inflammation - U 1286 [INFINITE]
Bouquet, L. [Auteur]
Guiot, M. [Auteur]
Venet, Fabienne [Auteur]
Hospices Civils de Lyon [HCL]
Hôpital Edouard Herriot [CHU - HCL]
Sujobert, P. [Auteur]
Roussel, X. [Auteur]
Rouzaire, Paul-Olivier [Auteur]
Role of intra-Clonal Heterogeneity and Leukemic environment in ThErapy Resistance of chronic leukemias [CHELTER]
CHU Clermont-Ferrand
Caillot, Denis [Auteur]
Service d'Hématologie Clinique (CHU de Dijon)
Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand [CHU Dijon]
Casasnovas, O. [Auteur]
Bories, Jean Christophe [Auteur]
Immunologie humaine, physiopathologie & immunothérapie [HIPI (UMR_S_976 / U976)]
Bachy, E. [Auteur]
Caillat-Zucman, S. [Auteur]
Deschamps, M. [Auteur]
Ferrand, C. [Auteur]

Titre de la revue :

Journal of Translational Medicine

Nom court de la revue :

J Transl Med

Numéro :

19

Pagination :

265

Date de publication :

2021-06-28

ISSN :

1479-5876

Mot(s)-clé(s) en anglais :

Monitoring
Axi-cel
Tisa-cel
IL-1RAP
Droplet digital PCR
Chimeric antigen receptor

Discipline(s) HAL :

Sciences du Vivant [q-bio]

Résumé en anglais : [en]

Background Genetically engineered chimeric antigen receptor (CAR) T lymphocytes are promising therapeutic tools for cancer. Four CAR T cell drugs, including tisagenlecleucel (tisa-cel) and axicabtagene-ciloleucel (axi-cel), ...
Lire la suite >Background Genetically engineered chimeric antigen receptor (CAR) T lymphocytes are promising therapeutic tools for cancer. Four CAR T cell drugs, including tisagenlecleucel (tisa-cel) and axicabtagene-ciloleucel (axi-cel), all targeting CD19, are currently approved for treating B cell malignancies. Flow cytometry (FC) remains the standard for monitoring CAR T cells using a recombinant biotinylated target protein. Nevertheless, there is a need for additional tools, and the challenge is to develop an easy, relevant, highly sensitive, reproducible, and inexpensive detection method. Molecular tools can meet this need to specifically monitor long-term persistent CAR T cells. Methods Based on 2 experimental CAR T cell constructs, IL-1RAP and CS1, we designed 2 quantitative digital droplet (ddPCR) PCR assays. By targeting the 4.1BB/CD3z (28BBz) or 28/CD3z (28z) junction area, we demonstrated that PCR assays can be applied to approved CD19 CAR T drugs. Both 28z and 28BBz ddPCR assays allow determination of the average vector copy number (VCN) per cell. We confirmed that the VCN is dependent on the multiplicity of infection and verified that the VCN of our experimental or GMP-like IL-1RAP CAR T cells met the requirement (< 5 VCN/cell) for delivery to the clinical department, similar to approved axi-cel or tisa-cel drugs. Results 28BBz and 28z ddPCR assays applied to 2 tumoral (acute myeloid leukemia (AML) or multiple myeloma (MM) xenograft humanized NSG mouse models allowed us to quantify the early expansion (up to day 30) of CAR T cells after injection. Interestingly, following initial expansion, when circulating CAR T cells were challenged with the tumor, we noted a second expansion phase. Investigation of the bone marrow, spleen and lung showed that CAR T cells disseminated more within these tissues in mice previously injected with leukemic cell lines. Finally, circulating CAR T cell ddPCR monitoring of R/R acute lymphoid leukemia or diffuse large B cell lymphoma (n = 10 for tisa-cel and n = 7 for axi-cel) patients treated with both approved CAR T cells allowed detection of early expansion, which was highly correlated with FC, as well as long-term persistence (up to 450 days), while FC failed to detect these events. Conclusion Overall, we designed and validated 2 ddPCR assays allowing routine or preclinical monitoring of early- and long-term circulating approved or experimental CAR T cells, including our own IL-1RAP CAR T cells, which will be evaluated in an upcoming phase I clinical trial.Lire moins >

Langue :

Anglais

Comité de lecture :

Oui

Audience :

Internationale

Vulgarisation :

Non

Établissement(s) :

Université de Lille
Inserm
CHU Lille

Collections :

• Institut de Recherche Translationnelle sur l'Inflammation (INFINITE) - U1286

Date de dépôt :

2024-01-12T06:37:54Z
2024-02-27T13:06:06Z