Search

Purpose: Although CD19 chimeric antigen receptor T cells (CAR-T) therapy has shown remarkable success in B-cell malignancies, a substantial fraction of patients do not obtain a long-term clinical response. This could be influenced by the quality of the individual CAR-T infusion product. To shed some light on this, clinical outcome was correlated to characteristics of CAR-T infusion products. Patients and Methods: In this phase II study, patients with B-cell lymphoma (n = 23) or leukemia (n = 1) received one or two infusions of third-generation CD19-directed CAR-Ts (2 × 108/m2). The clinical trial was registered at clinicaltrials.gov: NCT03068416. We investigated the transcriptional profile of individual CD19 CAR-T infusion products using targeted single-cell RNA sequencing and multicolor flow cytometry. Results: Two CAR-T infusions were not better than one in the settings used in this study. As for the CAR-T infusion products, we found that effector-like CD8+CAR-Ts with a high polyfunctionality, high cytotoxic and cytokine production profile, and low dysfunctional signature were associated with clinical response. An extended ex vivo expansion time during CAR-T manufacturing negatively influenced the proportion of effector CD8+CAR-Ts in the infusion product. Conclusions: We identified cell-intrinsic characteristics of effector CD8+CAR-Ts correlating with response that could be used as an indicator for clinical outcome. The results in the study also serve as a guide to CAR-T manufacturing practices., De två första författarna delar förstaförfattarskapetDe två sista författarna delar sistaförfattarskapet

In recent years, immunotherapy has revolutionized cancer treatment by prolonging survival and even curing patients lacking other available therapies. Besides immune checkpoint inhibitors, one of the major advances is attributed to the success of chimeric antigen receptor (CAR)-T cell therapy in treating patients with B-cell malignancies. Although many patients respond, some are resistant to treatment and others will relapse after an initial response. CAR-T cell efficacy can be enhanced by adding a transgene cassette encoding an immune-stimulatory molecule that induce bystander immunity and a broader anti-cancer response. This is particularly important to improve efficacy in solid tumors where the “CAR-target-antigen” is often heterogeneously expressed. The neutrophil-activating protein (NAP), a virulence factor of Helicobacter Pylori, is an efficient immune stimulator. When secreted from CAR-T cells NAP can induce bystander immunity with killing of “CAR-target-negative” tumor cells. In my thesis, I aim at investigating resistance mechanisms to CAR-T therapy and developing new improved CAR-T therapies. In paper I, clinical response in patients with B-cell lymphoma was evaluated after treatment with two doses of third-generation CD19CAR-T cells. Response was observed in 9 out of 24 patients (37.5%) irrespectively of receiving one or two doses. We also analyzed whether the profile of the individual CAR-T infusion products could predict response. CAR-Ts from responders had high cytotoxic and low dysfunctional profile while CAR-Ts from non-responders had high dysfunction profile. Extended culture time during manufacturing was associated with dysfunction. As CAR-T products of non-responders were on average cultured longer, this may explain their dysfunctionality. In paper II, we developed CD20-targeting CAR-T cells (CAR20-T), which efficiently killed CD20-expressing human lymphoma cell lines and patient-derived lymphoma cells in vitro. By arming CAR20-T cells with NAP we

Differentially expressed genes between responders and non-responders (Fc -1.2 ≤ or ≤ 1.2)

Stapled peptides targeting the interaction between p53 and its negative regulators MDM2 and MDM4 have exhibited great potential as anti-cancer drugs, albeit with room for improvement in formulation and tumor specificity. Lipid bilayer disks (lipodisks) have emerged as promising drug nanocarriers and can by attachment of targeting moieties be directed selectively towards tumor cells. Tumor-targeted delivery of stapled peptides by use of lipodisks may therefore increase the uptake in the tumors and limit toxicity in healthy tissue. Here, we utilized epidermal growth factor receptor (EGFR)-targeted lipodisks to deliver p53-activating stapled peptide VIP116 to EGFR-expressing tumor cells. We demonstrate that VIP116 can be stably formulated in lipodisks (maximum peptide/lipid molar ratio 0.11). In vitro cell studies verify specific binding of EGF-decorated lipodisks to tumor cells and confirm that targeted delivery of VIP116 significantly decreases tumor cell viability.