Chimeric antigen receptor cells have revolutionized the treatment of hematological malignancies. Anti-CD19 CAR T cells showed impressive results, reaching a complete response in 80% of cases in heavily pre-treated relapsing/ refractory B cell precursor acute lymphoblastic leukemia (r/r BPC-ALL). Despite these encouraging results, about 50% of treated patients eventually relapsed and 40% of relapses are CD19 negative, evading CAR T surveillance. Great efforts are directed to optimize CAR products to improve their efficacy and persistence. Our group developed a method based on the Sleeping Beauty transposon platform for CAR T cell engineering as a sustainable and flexible alternative to standard protocols exploiting viral vectors. Transposon vectors are safe and cheaper than viral vectors. Moreover, to favor access of lymphodepleted patients to CAR T immunotherapy, we exploited donor-derived Cytokine-induced killer (CIK) cells, a T cell population characterized by a higher cytotoxic activity and a lower risk of graft versus host disease (GvHD). Our anti-CD19 CARCIK product has been tested in a clinical trial for the treatment of patients with B-ALL relapsed after a hematopoietic stem cell transplant (NCT03389035). In this study, we proposed to prevent and treat CD19 negative relapses by a multitargeting strategy based on non-viral T cell engineering. In particular, we designed and cloned in Sleeping Beauty vectors second-generation anti-CD22 and anti-BAFF-R CARs. After testing the anti-tumor activity of the single CAR-modified CIK cells, we co-electroporated the two plasmids encoding CARs to generate a mixed anti-CD22 and anti-BAFF-R population of, including single and double targeting CAR T cells.
I recettori chimerici dell'antigene (CAR) hanno rivoluzionato il trattamento delle neoplasie ematologiche. Le cellule CAR T anti-CD19 hanno mostrato risultati impressionanti, permettendo di raggiungere tassi di risposta superiori all'80% anche in pazienti con leucemia linfoblastica acuta a cellule B resistente/refrattaria (B-LLA r/r) pesantemente pre-trattati. Nonostante questi risultati incoraggianti, circa il 50% dei pazienti trattati recidivano ed il 40% di queste recidive sono CD19 negative. Grandi sforzi sono stati diretti nell'ottimizzare la produzione dei CAR migliorandone l'efficacia e la persistenza. Il nostro gruppo ha sviluppato un metodo di ingegnerizzazione delle cellule CAR T basato su una piattaforma a trasposoni Sleeping Beauty, un metodo alternativo sostenibile e flessibile rispetto ai protocolli standard che utilizzano vettori virali. I vettori transposonici sono sicuri e più economici dei vettori virali. Inoltre, per favorire l'accesso all'immunoterapia con cellule CAR T dei pazienti linfopenici, abbiamo esplorato la possibilità di utilizzare cellule killer indotte da citochine (CIK) di derivazione da donatore, caratterizzate da una maggiore citotossicità ed un più basso rischio di sviluppare malattia del trapianto contro l'ospite. Il nostro prodotto anti-CD19 CAR-CIK è stato testato in un Clinical trial per il trattamento di pazienti con B-LLA recidivati dopo trapianto di cellule staminali ematopoietiche (NCT03389035). In questo studio, proponiamo per prevenire e trattare le recidive CD19 negative, una strategia di multi-targeting basata sull'ingegnerizzazione dei linfociti T tramite vettori non virali. In particolare abbiamo disegnato e clonato in vettori Sleeping Beauty due CAR di seconda generazione diretti rispettivamente contro il CD22 e il BAFF-R. Dopo aver testato la loro attività anti-tumorale in vitro, abbiamo co-elettroporato i due plasmidi codificanti i due CAR generando una popolazione mista composta di cellule CAR T esprimenti i singoli CAR od entrambi.
Anti-CD22 and Anti-BAFF-R CARCIK cells engineered with Sleeping Beauty transposon: towards B-ALL multitargeting strategy
MORETTI, ALEX
2024
Abstract
Chimeric antigen receptor cells have revolutionized the treatment of hematological malignancies. Anti-CD19 CAR T cells showed impressive results, reaching a complete response in 80% of cases in heavily pre-treated relapsing/ refractory B cell precursor acute lymphoblastic leukemia (r/r BPC-ALL). Despite these encouraging results, about 50% of treated patients eventually relapsed and 40% of relapses are CD19 negative, evading CAR T surveillance. Great efforts are directed to optimize CAR products to improve their efficacy and persistence. Our group developed a method based on the Sleeping Beauty transposon platform for CAR T cell engineering as a sustainable and flexible alternative to standard protocols exploiting viral vectors. Transposon vectors are safe and cheaper than viral vectors. Moreover, to favor access of lymphodepleted patients to CAR T immunotherapy, we exploited donor-derived Cytokine-induced killer (CIK) cells, a T cell population characterized by a higher cytotoxic activity and a lower risk of graft versus host disease (GvHD). Our anti-CD19 CARCIK product has been tested in a clinical trial for the treatment of patients with B-ALL relapsed after a hematopoietic stem cell transplant (NCT03389035). In this study, we proposed to prevent and treat CD19 negative relapses by a multitargeting strategy based on non-viral T cell engineering. In particular, we designed and cloned in Sleeping Beauty vectors second-generation anti-CD22 and anti-BAFF-R CARs. After testing the anti-tumor activity of the single CAR-modified CIK cells, we co-electroporated the two plasmids encoding CARs to generate a mixed anti-CD22 and anti-BAFF-R population of, including single and double targeting CAR T cells.File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.14242/172921
URN:NBN:IT:UNIMIB-172921