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Biology subjects

Mangolini, M.

Publications and source records attributed to Mangolini, M..

3 recordsLinked to original sources

Engineering armoured in vivo CAR T cells through targeted delivery and transient mRNA gating

In vivo generation of chimeric antigen receptor (CAR) T cells offers an off-the-shelf, scalable alternative to ex-vivo therapies, but is constrained by inefficiency, toxicity and durability. We report the first armoured in-vivo CAR T platform with tunable, transient activity, that enhances safety and efficacy. Our platform utilises CD8 targeted lipid nanoparticles (LNP) to transiently deliver CAR and armouring membrane-bound IL12 mRNAs. Payloads utilize a T-cell-restricted (T-trex) strategy to constrain CAR and IL-12 expression to T cells, enabling potent antigen-dependent activity with spatio-temporal control. Selective targeting of T-cells with a T-trex CAR showed enhanced expression. The addition of T-trex IL12 mRNA provides a tunable, localised amplification of CAR function, enhancing antigen-dependent cytotoxicity and cytokine production with robust tumour control. These findings establish a novel dose-efficient armoured in-vivo CAR T approach and general framework for programmable immune cell engineering.

immunology↗

Highly potent novel multi-armoured IL13Rα2 CAR-T subverts the immunosuppressive microenvironment of Glioblastoma

Glioblastoma remains one of the most challenging and lethal brain cancers, with limited treatment options. While CAR-T cells have shown promise in some patients, sustaining T-cell activity and overcoming the immunosuppressive tumour microenvironment remain significant hurdles. Here, we present an armoured CAR-T cell design to address these challenges and enhance persistence in GBM tumours. We developed a highly specific humanised single-domain antibody targeting IL13R2 and included it alongside four additional modular elements in a single retroviral vector for CAR-T generation. Our results demonstrate that this single-cassette CAR-T cell design possesses high resilience against TGF-{beta}-mediated immunosuppression, enhanced tumour-killing capacity through IL-12 secretion while maintaining a favourable safety profile, extended persistence in the host, and an additional layer of safety control through the incorporation of a suicide switch. Importantly, despite its complexity, the construct can still be manufactured efficiently. These advancements represent a significant step forward in addressing key challenges associated with CAR-T cell therapy in solid tumours.

cancer biology↗

NOTCH1 drives immune-escape mechanisms in B cell malignancies

NOTCH1 is a recurrently mutated gene in Chronic Lymphocytic Leukemia (CLL) and Mantle Cell Lymphoma (MCL). Functional studies to investigate its role have been hampered by the inability to genetically manipulate primary human lymphoma cells, attributed to low transduction-efficacy and procedure-associated toxicity. To overcome these limitations, we have developed a novel method to retrovirally transfer genes into malignant human B cells. We generated isogenic human tumor cells from patients with CLL and MCL, differing only in their expression of NOTCH1. Our data demonstrate that NOTCH1 facilitates immune-escape of malignant B cells by up-regulating PD-L1, partly dependent on autocrine interferon-{gamma} signaling. In addition, NOTCH1 causes silencing of the entire HLA-class II locus via suppression of the transcriptional co-activator CIITA. These NOTCH1-mediated immune escape mechanisms are associated with the expansion of CD4+ T cells in vivo, further contributing to the poor clinical outcome of NOTCH1-mutated CLL and MCL.

cancer biology↗