Project description:Background: T cells expressing antigen-specific chimeric antigen receptors (CARs) improve outcomes for CD19-expressing B-cell malignancies. We provide the first human application of T cells genetically modified using the Sleeping Beauty (SB) transposon/transposase system to express a CD19-specific CAR. Methods: T cells were genetically modified using DNA plasmids from the SB platform to stably express a second-generation CD19-specific CAR and selectively propagated ex vivo with activating and propagating cells (AaPC) and cytokines. Twenty-six patients with advanced NHL and ALL safely underwent hematopoietic stem-cell transplantation (HSCT) and infusion of CAR T cells as adjuvant therapy in the autologous (n=7) or allogeneic settings (n=19). Results: SB-mediated genetic transposition and stimulation resulted in 2,200-2,500-fold ex vivo genetically modified T-cell expansion, with 84% CAR expression, and without integration hotspots. The 30-month progression-free and overall survivals were 83% and 100%, respectively, following autologous HSCT, and the respective 12-month rates after allogeneic HSCT were 53% and 63%. No acute or late toxicities and no exacerbation of graft-versus-host-disease were observed. Despite a low antigen burden and unsupportive recipient cytokine environment, CAR T cells persisted for an average of 201 days for autologous and 51 days for allogeneic recipients. Conclusions: CD19-specific CAR T cells generated with SB and AaPC platforms were safe, and may provide additional cancer control as planned infusions after HSCT, supporting further clinical development of this non-viral gene therapy approach.
Project description:Long-Term Follow-Up Study for Subjects Enrolled in the Phase I/II Study of Autologous T Cells Engineered using the Sleeping Beauty System to Express T cell Receptors (TCRs) Reactive Against Cancer-specific Mutations in Subjects with Solid Tumors
Project description:Thymic lymphomas were generated by inducing Sleeping Beauty transposon mutagenesis at different stages of T-cell development. This dataset includes exon array results from 14 tumor samples from two different Sleeping Beauty models of T-ALL (7 Vav-SB and 7 CD4-SB samples).
Project description:A Phase I/II study of autologous T cells engineered using the Sleeping Beauty transposon/transposase system to express TCR(s) reactive against neoantigens in subjects with relapsed/refractory solid tumors
Project description:We successfully established a novel immunocompetent mouse model of CHB(chronic hepatitis B) at the immune-active phase by a simple combination of a Fumarylacetoacetate hydrolase (FAH) knockout mouse, hydrodynamic tail vein injection, and sleeping-beauty transposon system. The employment of single-cell RNA sequence in this model revealed the comprehensive intrahepatic immune landscape in the immune-active phase, which contributes to the persistence of HBV infection. Our model also made it possible to profile the intrahepatic immune response to the immunomodulators.
Project description:A protein aggregation model of human ataxin-1(Q82) was generated in human mesenchymal cells using the Sleeping Beauty transposon system. These cells inducibly express the pathogenic protein that causes SCA1 disease. Here, we describe the transcriptional changes occurring in cells which contain different types of ataxin-1(Q82) protein inclusions
