Tuning charge density of chimeric antigen receptor optimizes tonic signaling and CAR-T cell fitness

Chen, Jian; Qiu, Shizhen; Li, Wentao; Wang, Kun; Yang, Han Kwang; Liu, Baichuan; Li, Guangfei; Li, Li; Chen, Min; Lan, Junjie; Niu, Jun; He, Peng; Cheng, Lei; Fan, Guang; Liu, Xin; Song, Xianmin; Xu, Chenqi; Wu, Haitao; Wang, Haopeng

doi:10.1038/s41422-023-00789-0

Cited by 70 publications

(47 citation statements)

References 59 publications

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“…We identified a few IDRs, including those from FUS, EWS, and TAF15, that promote the membrane-proximal signaling, cytotoxic factor production, and killing of CAR-Ts against low antigen-expressing cancers (CD19, CD22, and HER2). Moreover, no elevated tonic signaling was observed in IDR CAR-Ts, suggesting a difference in signaling outcomes between IDRinduced condensation and previously reported CAR aggregation (Chen et al, 2023;Long et al, 2015). Together, these results demonstrated that IDRs, though not originally linked to T cell signaling, can serve as a new modular motif to improve the anti-tumor effect of CAR-Ts.…”

Section: Introductionmentioning

confidence: 53%

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IDR-induced CAR condensation improves the cytotoxicity of CAR-Ts against low-antigen cancers

Zhang,

Xiao,

Zeng

et al. 2023

Preprint

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SummaryChimeric antigen receptor (CAR)-T cell-based therapies demonstrate remarkable efficacy for the treatment of otherwise intractable cancers, particularly B-cell malignancies. However, CAR-T cells respond to only a subset of cancers and are limited by low signaling efficiency: a few hundred or more antigen molecules are required to activate a CAR T cell whereas a single peptide-loaded MHC molecule is sufficient to trigger the activation of a normal T cell. This low antigen sensitivity not only limits the current CAR-T therapy to high antigen-expressing cancers, but also results in high frequencies of relapse, during which high antigen-expressing cancers downregulate their antigen levels to escape CAR-T’s attacking. To improve the antigen sensitivity of CAR-T cells, we engineered CARs targeting CD19, CD22, and HER2 by including intrinsically disordered regions (IDRs) that promote signaling condensation. The “IDR CARs” triggered enhanced membrane-proximal signaling in the CAR-T synapse, which led to an increased release of cytotoxic factors, a higher killing activity towards low antigen-expressing cancer cells in vitro, and an improved anti-tumor efficacy in vivo. No elevated tonic signaling was observed in IDR CAR-Ts. Together, we demonstrated IDRs as a new tool set to enhance CAR-T cytotoxicity and broaden CAR-T’s application to low antigen-expressing cancers.

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Section: Introductionmentioning

confidence: 53%

“…Previous work showed that aggregation of CARs targeting GD2 or CSPG4 induces tonic signaling, which triggers CAR-T activation in the absence of antigens (Chen et al, 2023; Long et al, 2015). Therefore, we assessed if IDR-induced CAR condensation causes tonic signaling.…”

Section: Resultsmentioning

confidence: 99%

IDR-induced CAR condensation improves the cytotoxicity of CAR-Ts against low-antigen cancers

Zhang,

Xiao,

Zeng

et al. 2023

Preprint

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“…It was found that CAR molecules with more positively charged amino acid residues, or positively charged patches (PCP), in scFv are more likely to aggregate spontaneously and generate stronger electrostatic field and substrate signals. When the ionic strength in the environment is enhanced by increasing the concentration of sodium chloride in the enhanced CAR‐T medium, the aggregation of CAR molecules is effectively prevented and the tonic signal is significantly reduced 49 . After mutating several positively charged residues in the scFv framework region (FR) of a CAR molecule with large PCP to uncharged residues, the CAR‐T cells showed a significant reduction of tonic signaling.…”

Section: Car Tonic Signaling Is Determined By the Surface Charge Of T...mentioning

confidence: 99%

“…CAR‐T cells overexpressing the reduced positively charged CAR molecules were less exhausted and showed enhanced antitumor efficacy in vivo. Importantly, a metric to quantify positively charged patches on the CAR surface, the PCP score, was defined and can be used as a reliable predictor of tonic signaling of CAR molecules 49 . This study brings new design ideas to the CAR‐T field, in which the PCP score on the surface needs to be considered while constructing more powerful CARs.…”

Section: Car Tonic Signaling Is Determined By the Surface Charge Of T...mentioning

confidence: 99%

“…We propose a “Peak Theory” highlighting that optimal CAR signaling is a key of clinical efficacy of CAR‐T cell therapy (Figure 3C). This theory has been justified for abovementioned CAR tonic signaling and needs to be further proven for antigen‐induced CAR signaling 49 . For CARs with high tonic signaling (e.g., GD2.CAR and CSPG4.CAR), reducing PCPs on CARs or enhancing ionic strength in the culture medium during ex vivo CAR‐T expansion minimized spontaneous activation and alleviated CAR‐T cell exhaustion.…”

Section: The Peak Model: Achieving Optimal Signaling For Effective Ca...mentioning

confidence: 99%

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Charging CAR by electrostatic power

Wang

Huang

2023

Immunological Reviews

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SummaryChimeric antigen receptor (CAR)‐T cell therapy has emerged as a promising approach for cancer treatment. CAR is a synthetic immune receptor that recognizes tumor antigen and activates T cells through multiple signaling pathways. However, the current CAR design is not as robust as T cell receptor (TCR), a natural antigen receptor with high sensitivity and efficiency. TCR signaling relies on specific molecular interactions, and thus electrostatic force, the major force of molecular interactions, play critical roles. Understanding how electrostatic charge regulates TCR/CAR signaling events will facilitate the development of next‐generation T cell therapies. This review summarizes recent findings on the roles of electrostatic interactions in both natural and synthetic immune receptor signaling, specifically that in CAR clustering and effector molecule recruitments, and highlights potential strategies for engineering CAR‐T cell therapy by leveraging charge‐based interactions.

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Protein tyrosine phosphatases: emerging role in cancer therapy resistance

Zhao,

Shuai,

et al. 2024

Cancer Communications

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BackgroundTyrosine phosphorylation of intracellular proteins is a post‐translational modification that plays a regulatory role in signal transduction during cellular events. Dephosphorylation of signal transduction proteins caused by protein tyrosine phosphatases (PTPs) contributed their role as a convergent node to mediate cross‐talk between signaling pathways. In the context of cancer, PTP‐mediated pathways have been identified as signaling hubs that enabled cancer cells to mitigate stress induced by clinical therapy. This is achieved by the promotion of constitutive activation of growth‐stimulatory signaling pathways or modulation of the immune‐suppressive tumor microenvironment. Preclinical evidences suggested that anticancer drugs will release their greatest therapeutic potency when combined with PTP inhibitors, reversing drug resistance that was responsible for clinical failures during cancer therapy.Areas coveredThis review aimed to elaborate recent insights that supported the involvement of PTP‐mediated pathways in the development of resistance to targeted therapy and immune‐checkpoint therapy.Expert opinionThis review proposed the notion of PTP inhibition in anticancer combination therapy as a potential strategy in clinic to achieve long‐term tumor regression. Ongoing clinical trials are currently underway to assess the safety and efficacy of combination therapy in advanced‐stage tumors.

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Tuning charge density of chimeric antigen receptor optimizes tonic signaling and CAR-T cell fitness

Cited by 70 publications

References 59 publications

IDR-induced CAR condensation improves the cytotoxicity of CAR-Ts against low-antigen cancers

IDR-induced CAR condensation improves the cytotoxicity of CAR-Ts against low-antigen cancers

Charging CAR by electrostatic power

Protein tyrosine phosphatases: emerging role in cancer therapy resistance

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