HLA-independent T cell receptors for targeting tumors with low antigen density

Mansilla-Soto, Jorge; Eyquem, Justin; Haubner, Sascha; Hamieh, Mohamad; Feucht, Judith; Paillon, Noémie; Zucchetti, Andrés E.; Li, Zhuoning; Sjöstrand, Maria; Lindenbergh, Pieter L.; Saetersmoen, Michelle; Dobrin, Anton; Maurin, Mathieu; Iyer, Archana; Angus, Andreina Garcia; Miele, Matthew M.; Zhao, Zeguo; Giavridis, Theodoros; Stegen, Sjoukje J. C. van der; Tamzalit, Fella; Rivière, Isabelle; Huse, Morgan; Hendrickson, Ronald C.; Hivroz, Claire; Sadelain, Michel

doi:10.1038/s41591-021-01621-1

Cited by 106 publications

(88 citation statements)

References 59 publications

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“…While we have focused on cell proliferation as measured by abundance in this study, pooled knockin screens can be adapted to assess more complex phenotypes such as cytokine release or T cell infiltration into a tumor in vivo and will be a crucial tool in the discovery of synthetic genetic modifications that can be engineered to specifically enhance T cell identity and behavior across therapeutic contexts. In the future, modular pooled knockin screens should be readily adaptable to discover constructs that improve function of different CARs or TCRs and even newer synthetic antigen receptors such as HITs (HLA-independent TCRs) (Mansilla-Soto et al, 2022), STARs (synthetic T cell receptor and antigen receptors) (Liu et al, 2021b) or SNIPRs (synthetic intramembrane proteolysis receptors) and SynNotch receptors (Hyrenius-Wittsten et al, 2021; Zhu et al, 2022)). Furthermore, future screens can be performed in regulatory T cells (Tregs) to facilitate the development of treatments for autoimmunity or inflammatory diseases, or in gamma delta T cells.…”

Section: Discussionmentioning

confidence: 99%

Modular Pooled Discovery of Synthetic Knockin Sequences to Program Durable Cell Therapies

Blaeschke

Chen

Apathy

et al. 2022

Preprint

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Chronic stimulation can cause T cell dysfunction and limit efficacy of cellular immunotherapies. CRISPR screens have nominated gene targets for engineered T cells, but improved methods are required to compare large numbers of synthetic knockin sequences to reprogram cell functions. Here, we developed Modular Pooled Knockin Screening (ModPoKI), an adaptable platform for modular construction of DNA knockin libraries using barcoded multicistronic adaptors. We built two ModPoKI libraries of 100 transcription factors (TFs) and 129 natural and synthetic surface receptors. Over 20 ModPoKI screens across human TCR and CAR T cells in diverse conditions identified a transcription factor AP4 (TFAP4) construct to enhance long-term T cell fitness and anti-cancer function in vitro and in vivo. ModPoKI's modularity allowed us to generate a ~10,000-member library of TF combinations. Non-viral knockin of a combined BATF-TFAP4 polycistronic construct further enhanced function in vivo. ModPoKI facilitates discovery of complex gene constructs to program cellular functions.

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Section: Discussionmentioning

confidence: 99%

Modular Pooled Discovery of Synthetic Knockin Sequences to Program Durable Cell Therapies

Blaeschke

Chen

Apathy

et al. 2022

Preprint

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“…A second approach to improving CAR function has focused on exploiting all the signalling domains present in the TCR/CD3 complex. For example, eTruC receptors fuse the scFv directly to the extracellular domain of CD3 ε (19) whereas STARs (also called HIT receptors) replace the variable domains of the TCR with the scFv variable domains (20, 21). Using a xenograft carcinoma model with EGFR as the target antigen a STAR outperformed an eTruC, and both outperformed CARs (20).…”

Section: Introductionmentioning

confidence: 99%

Inefficient exploitation of accessory receptors reduces the sensitivity of chimeric antigen receptors

Burton

Siller-Farfán

Pettmann

et al. 2021

Preprint

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Chimeric antigen receptors (CARs) can re-direct T cells to target abnormal cells but their activity is limited by a profound defect in antigen sensitivity, the source of which remains unclear. Here, we show that CARs have a >100-fold lower antigen sensitivity compared to the TCR when antigen is presented on antigen-presenting-cells, but nearly identical sensitivity when antigen is presented as purified protein in isolation. Given that the TCR uses other, accessory, receptors to achieve high sensitivity, we screened prominent accessory receptors by presenting their purified ligands together with antigen. We found that ligating the adhesion receptor CD2 or LFA-1 improved antigen sensitivity for the TCR by >100-fold, whereas for CARs the improvement was <10-fold. We reproduced these findings using target cells where the CD2 and/or LFA-1 interaction were abrogated. Sensitivity can be partially restored by fusing the CAR variable domains to the TCR CD3ε subunit (also known as a TRuC) and fully restored when exchanging the TCRαβ variable domains for those of the CAR (also known as a STAR). Our study localises the defect in CAR sensitivity to inefficient use of accessory receptors and suggests approaches to increase sensitivity.

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“…Immune escape by tumors can occur through multiple mechanisms such as antigen loss or binding escaping mutations within the same antigen [34, 35]. Multispecific CAR-T cells potentntly eliminate antigen-heterogeneous B cell tumors in preclinical models to conquer the relapse caused by antigen loss or down-regulation [36].…”

Section: Discussionmentioning

confidence: 99%

“…Optimization of the CAR construct can further increase the efficacy. In one recent study, a CD19 specific CAR was designed inside T cell receptor-CD3 complex targeting cell surface antigens of low abundance (<20 antigens per cell) [35]. We are testing our 3C9 CAR-Ts based on the similar strategy.…”

Section: Discussionmentioning

confidence: 99%

Discovery of a novel human antibody VH domain with potent activity against mesothelin expressing cancer cells in both CAR T-cell and antibody drug conjugate formats

Sun

Chu

Adams

et al. 2022

Preprint

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Antibody based therapeutics targeting mesothelin (MSLN) have shown limited anti-tumor activities in clinical trials. Novel antibodies with high affinity and better therapeutic developability properties are needed as preclinical candidates. In the present study, we have isolated and characterized a novel VH domain 3C9 from a large size human immunoglobulin heavy chain variable (VH) domain library. 3C9 exhibited high affinity [KD (dissociation constant) < 3nM] and demonstrated good specificity in a membrane proteome array (MPA). Both CAR-T cells and antibody domain drug conjugations (DDCs) generated with 3C9, showed effective killing of MSLN positive cells in vitro without off-target effects. This is also the first report of the crystal structure of MSLN based on the analysis of the MSLN-3C9 complex which was solved at 2.9 A resolution. The newly identified antibody domain is a promising candidate therapeutic against cancer.

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HLA-independent T cell receptors for targeting tumors with low antigen density

Cited by 106 publications

References 59 publications

Modular Pooled Discovery of Synthetic Knockin Sequences to Program Durable Cell Therapies

Modular Pooled Discovery of Synthetic Knockin Sequences to Program Durable Cell Therapies

Inefficient exploitation of accessory receptors reduces the sensitivity of chimeric antigen receptors

Discovery of a novel human antibody VH domain with potent activity against mesothelin expressing cancer cells in both CAR T-cell and antibody drug conjugate formats

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