Reversal of Remimazolam-Mediated Inhibition of Jurkat T Cell Activation by TGFBI Depletion

Abstract

GABA_A receptors are well-recognized targets for intravenous anesthetics and have been identified in T lymphocytes. Remimazolam, a GABA_A receptor-binding agent, enhances the inhibitory effects of γ-aminobutyric acid (GABA) and provides a rapid onset and offset of sedation, making it suitable for procedural sedation and anesthesia. However, the impact of remimazolam on T cell function remains poorly understood. In this study, we used mass spectrometry analysis to confirm that Jurkat T cells produce and secrete GABA de novo. Consequently, treatment with remimazolam inhibited Jurkat T cell activation, even in the absence of exogenous GABA. Transcriptomic profiling of remimazolam-treated Jurkat T cells exhibited a significant upregulation of TGFBI expression. Furthermore, CRISPR/Cas9-mediated knockout of TGFBI reversed the inhibitory effects of remimazolam on Jurkat T cell activation. These findings highlight the profound influence of anesthetics on T cell activation and could be crucial for optimizing their clinical application.

Keywords

GABA_A receptor / GABA / Remimazolam / T cell activation / TGFBI

Supplementary Materials

The following supporting information can be found at: https://www.sciepublish.com/article/pii/625. Figure S1. Gate strategies for the analysis of CD69 expression and p-ERK levels in WT or TGFBI^−/− Jurkat T cells following various treatments. (A) Gate strategies for the analysis of CD69 expression on Jurkat T cells following various treatments. (B) Gate strategies for the analysis of CD69 expression on WT and TGFBI^−/− Jurkat T cells in a steady state. (C) Gate strategies for the analysis of CD69 expression on WT and TGFBI^−/− Jurkat T cells following P+I treatment. (D) Gate strategies for the analysis of CD69 expression on WT and TGFBI^−/− Jurkat T cells following P+I+Remi treatment. (E,F) Gate strategies for the analysis of p-ERK levels in WT and TGFBI^−/− Jurkat T cells following DMSO (NT) or P+I treatments. (G) Gate strategies for the analysis of CD69 expression on TGFBI^−/− Jurkat T cells following P+I or P+I+Remi treatment. Figure S2. Knockout of TGFBI reduces GABA production by Jurkat T cells. (A) Mass spectrometry detection of GABA in WT and TGFBI knockout Jurkat T cells (up) and their corresponding culture medium following stimulation with or without P+I. (B) Quantification of GABA levels in WT and TGFBI deficiency Jurkat T cells and culture medium following stimulation with or without P+I. Statistical significance is denoted as follows: ** p < 0.01, **** p < 0.0001, unpaired Student’s t-test.

Author Contributions

Y.L. (Yinming Liang) and H.W. (Hui Wang) contributed conception and design of the study. B.Z. (Binhui Zhou) and Y.L. (Yinming Liang) wrote the manuscript; H.W. (Hao Wang), G.G., H.M., Y.Q., B.Z. (Bowen Zhang), T.L. (Tingting Liu), Y.L. (Yang Liu), T.L. (Tianhan Li), and L.Z. contributed to the experiments. All authors contributed to the manuscript revision, and have read and approved the submitted version.

Ethics Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data will be made available on request.

Funding

This work was supported by The Natural Science Foundation of Henan Province to Yinming Liang (grant number.222300420015) and the start-up fund from Xinxiang Medical University to Binhui Zhou (grant number. 505483).

Declaration of Competing Interest

The authors declare no commercial or financial conflict of interest.

References

[1] Saxena NC, Macdonald RL. Properties of putative cerebellar gamma-aminobutyric acid A receptor isoforms. Mol. Pharmacol. 1996, 49, 567-579.

[2] Mendu SK, Bhandage A, Jin Z, Birnir B. Different subtypes of GABA-A receptors are expressed in human, mouse and rat T lymphocytes. PLoS ONE 2012, 7, e42959.

[3] Garcia PS, Kolesky SE, Jenkins A. General anesthetic actions on GABA(A) receptors. Curr. Neuropharmacol. 2010, 8, 2-9.

[4] Dionisio L, Arias V, Bouzat C, Esandi Mdel C. GABAA receptor plasticity in Jurkat T cells. Biochimie 2013, 95, 2376-2384.

[5] Sparrow EL, James S, Hussain K, Beers SA, Cragg MS, Bogdanov YD. Activation of GABA(A) receptors inhibits T cell proliferation. PLoS ONE 2021, 16, e0251632.

[6] Barragan A, Weidner JM, Jin Z, Korpi ER, Birnir B. GABAergic signalling in the immune system. Acta Physiol. 2015, 213, 819-827.

[7] Watanabe M, Maemura K, Kanbara K, Tamayama T, Hayasaki H. GABA and GABA receptors in the central nervous system and other organs. Int. Rev. Cytol. 2002, 213, 1-47.

[8] Kilpatrick GJ, McIntyre MS, Cox RF, Stafford JA, Pacofsky GJ, Lovell GG, et al. CNS 7056: a novel ultra-short-acting Benzodiazepine. Anesthesiology 2007, 107, 60-66.

[9] Korol SV, Jin Z, Jin Y, Bhandage AK, Tengholm A, Gandasi NR, et al. Functional characterization of native, high-affinity GABAA receptors in human pancreatic β cells. EBioMedicine 2018, 30, 273-282.

[10] Kang S, Liu L, Wang T, Cannon M, Lin P, Fan TWM, et al. GAB functions as a bioenergetic and signalling gatekeeper to control T cell inflammation. Nat. Metab. 2022, 4, 1322-1335.

[11] Lee A, Shirley M. Remimazolam: a review in procedural sedation. Drugs 2021, 81, 1193-1201.

[12] Zhang H, Li H, Zhao S, Bao F. Remimazolam in general anesthesia: A comprehensive review of its applications and clinical efficacy. Drug Des. Devel. Ther. 2024, 18, 3487-3498.

[13] Hu Q, Liu X, Wen C, Li D, Lei X. Remimazolam: an updated review of a new sedative and anaesthetic. Drug Des. Dev. Ther. 2022, 16, 3957-3974.

[14] Wang Z, Bian L, Mo C, Shen H, Zhao LJ, Su K-J, et al. Quantification of aminobutyric acids and their clinical applications as biomarkers for osteoporosis. Communications Biology 2020, 3, 39.

[15] Cai J, Zhu L, Zha Y, Kang Q. TGFBI gene mutation analysis in chinese families with corneal dystrophies. Genet. Test. Mol. Biomarkers 2016, 20, 388-392.

[16] Yu H, Wergedal JE, Zhao Y, Mohan S. Targeted disruption of TGFBI in mice reveals its role in regulating bone mass and bone size through periosteal bone formation. Calcif. Tissue Int. 2012, 91, 81-87.

[17] Corona A, Blobe GC. The role of the extracellular matrix protein TGFBI in cancer. Cell Signal. 2021, 84, 110028.

[18] Peng P, Zhu H, Liu D, Chen Z, Zhang X, Guo Z, et al. TGFBI secreted by tumor-associated macrophages promotes glioblastoma stem cell-driven tumor growth via integrin αvβ5-Src-Stat3 signaling. Theranostics 2022, 12, 4221-4236.

[19] Guo S-K, Shen M-F, Yao H-W, Liu Y-S. Enhanced expression of TGFBI promotes the proliferation and migration of glioma cells. Cell Physiol. Biochem. 2018, 49, 1138-1150.

[20] Wu L, Jiang L, Zhou Y, Zheng W, Feng A, Guo H. TGFBI regulates the TGF-β pathway to affect the malignant progression and cisplatin sensitivity in diffuse large B-cell lymphoma. Ann. Hematol. 2025, 104, 1165-1176.