LRRC8A:C/E Heteromeric Channels Are Ubiquitous Transporters of cGAMP

Lahey, Lauren J.; Mardjuki, Rachel E.; Wen, Xianlan; Hess, Gaelen T.; Ritchie, Christopher; Carozza, Jacqueline A.; Böhnert, Volker; Maduke, Merritt; Bassik, Michael C.; Li, Lingyin

doi:10.1016/j.molcel.2020.10.021

Cited by 125 publications

(146 citation statements)

References 81 publications

(123 reference statements)

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“…55 The bacterial CDNs 3'3'-cGAMP and 3'3'-CDA are associated with pathogenic bacteria, [56][57][58] while 3'3'-CDG is produced by a wide variety of bacteria, including commensals. 59 The ability of SLC46A2 and other CDN transporters 16,18 to selectively import certain CDNs (such as cGAMP and 3'3'-CDA) but not others (3'3'-CDG) suggests that CDN transporters could regulate how the immune system differentially responds to pathogenic and commensal bacteria.…”

Section: Discussionmentioning

confidence: 99%

“…16 Despite this, another inhibitor of SLC19A1, sulfasalazine (SSZ), strongly inhibited extracellular cGAMP signaling in CD14 + monocytes as determined by IRF3 phosphorylation (Figure 1A). We previously identified the LRRC8A channels as broadly expressed cGAMP transporters; 18 however, SSZ is not known to inhibit LRRC8A channels. This suggests that SSZ is inhibiting an unknown cGAMP transporter in CD14 + monocytes.…”

Section: Cd14 + Monocytes Express High Levels Of the Uncharacterized Transporter Slc46a2mentioning

confidence: 99%

“…Custom plasmids (pTwist-CMV) containing the coding sequences of mouse Slc19a1, Slc46a1, Slc46a2, and Slc46a3 were purchased from Twist Bioscience. To generate doxycycline inducible lentiviral plasmids, the transporter CDS was amplified from the appropriate plasmid using the primers listed in Supplementary Table 1 and cloned into EcoRI/BamHI linearized pLVX-TetOne-FLAG-Hydro plasmid 18 by isothermal Gibson assembly. 68 To create a construct encoding SLC46A2 with an extracellular loop FLAG tag (pTwist-SLC46A2-exFLAG), a custom plasmid containing the codon optimized CDS of SLC46A2 (pTwist-SLC46A2) was linearized with AfeI and an oligonucleotide encoding a GGSG-linker flanked FLAG tag (Supplementary Table 1) was ligated into the cut site.…”

Section: Recombinant Dnamentioning

confidence: 99%

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Human SLC46A2 is the dominant cGAMP importer in extracellular cGAMP-sensing macrophages and monocytes

Cordova

Ritchie

Böhnert

et al. 2020

Preprint

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Administration of exogenous CDNs to activate the cGAMP-STING pathway is a promising therapeutic strategy to unleash the full potential of cancer immunotherapy. This strategy mirrors the role of endogenous extracellular cGAMP, which we recently described as an immunotransmitter exported by cancer cells and imported into local responder cells of unknown identities to promote anti-tumoral immunity, with irradiation enhancing this effect. Here, in low-dose irradiated murine tumors, we identified CD4 + T cells, M1 macrophages, and NKG2D Low NK cells as cGAMP responder cells that have decreased STING activation upon depletion of extracellular cGAMP. At higher doses of radiation, extracellular cGAMP promoted the death of T cells and macrophages. Furthermore, we identified the orphan protein SLC46A2 as the dominant importer of cGAMP and select bacterial CDNs in human macrophages and monocytes. Together, we provide the first cellular and molecular mechanisms of cGAMP as an immunotransmitter, paving the way for effective STING pathway therapeutics.

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Section: Discussionmentioning

confidence: 99%

Section: Cd14 + Monocytes Express High Levels Of the Uncharacterized Transporter Slc46a2mentioning

confidence: 99%

Section: Recombinant Dnamentioning

confidence: 99%

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Human SLC46A2 is the dominant cGAMP importer in extracellular cGAMP-sensing macrophages and monocytes

Cordova

Ritchie

Böhnert

et al. 2020

Preprint

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“… 68 Although asymmetric, 2′3′-cGAMP was efficiently transferred inside or outside of cells, which was largely mediated by various characterized cGAMP importer SLC19A1 181 , 182 or transporter LRRC8A:C/E heteromeric channels. 183 , 184 Increased expression of SLC19A1 was observed in SLE (systemic lupus erythematosus) patients 185 that might promote innate immunity activation. 2′3′-cGAMP was found to be steadily released from cells 186 but degraded by extracellular ENPP1 (ecto-nucleotide pyrophosphatase/phosphodiesterase).…”

Section: Regulation Of the Cgas Enzymatic Product 2′3′-cgampmentioning

confidence: 99%

Cytosolic DNA sensing by cGAS: regulation, function, and human diseases

Liu

2021

Sig Transduct Target Ther

118

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Sensing invasive cytosolic DNA is an integral component of innate immunity. cGAS was identified in 2013 as the major cytosolic DNA sensor that binds dsDNA to catalyze the synthesis of a special asymmetric cyclic-dinucleotide, 2′3′-cGAMP, as the secondary messenger to bind and activate STING for subsequent production of type I interferons and other immune-modulatory genes. Hyperactivation of cGAS signaling contributes to autoimmune diseases but serves as an adjuvant for anticancer immune therapy. On the other hand, inactivation of cGAS signaling causes deficiency to sense and clear the viral and bacterial infection and creates a tumor-prone immune microenvironment to facilitate tumor evasion of immune surveillance. Thus, cGAS activation is tightly controlled. In this review, we summarize up-to-date multilayers of regulatory mechanisms governing cGAS activation, including cGAS pre- and post-translational regulations, cGAS-binding proteins, and additional cGAS regulators such as ions and small molecules. We will also reveal the pathophysiological function of cGAS and its product cGAMP in human diseases. We hope to provide an up-to-date review for recent research advances of cGAS biology and cGAS-targeted therapies for human diseases.

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“…Notably, cGAS-mediated detection of VACV leads to the production of cGAMP that could be efficiently transferred to bystander cells, triggering the activation of a STING-dependent antiviral immunity in non-infected cells (65). Different models of cell-to-cell transfer of cGAMP have been proposed occurring through extracellular vesicles such as exosomes (66), gap-junctions (67,68), and incorporation into enveloped viruses (69) in addition to the recently described cGAMP transporters (70)(71)(72). Importantly, cGAS was rapidly described as a main sensor of HIV and other retroviruses (73).…”

Section: Cgas: a Main Actor Of Cellular Response To Dna And Rna Virusesmentioning

confidence: 99%

Cyclic Guanosine Monophosphate–Adenosine Monophosphate Synthase (cGAS), a Multifaceted Platform of Intracellular DNA Sensing

Verrier

Langevin

2021

Front. Immunol.

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Innate immune pathways are the first line of cellular defense against pathogen infections ranging from bacteria to Metazoa. These pathways are activated following the recognition of pathogen associated molecular patterns (PAMPs) by membrane and cytosolic pattern recognition receptors. In addition, some of these cellular sensors can also recognize endogenous danger-associated molecular patterns (DAMPs) arising from damaged or dying cells and triggering innate immune responses. Among the cytosolic nucleic acid sensors, the cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) synthase (cGAS) plays an essential role in the activation of the type I interferon (IFNs) response and the production of pro-inflammatory cytokines. Indeed, upon nucleic acid binding, cGAS synthesizes cGAMP, a second messenger mediating the activation of the STING signaling pathway. The functional conservation of the cGAS-STING pathway during evolution highlights its importance in host cellular surveillance against pathogen infections. Apart from their functions in immunity, cGAS and STING also play major roles in nuclear functions and tumor development. Therefore, cGAS-STING is now considered as an attractive target to identify novel biomarkers and design therapeutics for auto-inflammatory and autoimmune disorders as well as infectious diseases and cancer. Here, we review the current knowledge about the structure of cGAS and the evolution from bacteria to Metazoa and present its main functions in defense against pathogens and cancer, in connection with STING. The advantages and limitations of in vivo models relevant for studying the cGAS-STING pathway will be discussed for the notion of species specificity and in the context of their integration into therapeutic screening assays targeting cGAG and/or STING.

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LRRC8A:C/E Heteromeric Channels Are Ubiquitous Transporters of cGAMP

Cited by 125 publications

References 81 publications

Human SLC46A2 is the dominant cGAMP importer in extracellular cGAMP-sensing macrophages and monocytes

Human SLC46A2 is the dominant cGAMP importer in extracellular cGAMP-sensing macrophages and monocytes

Cytosolic DNA sensing by cGAS: regulation, function, and human diseases

Cyclic Guanosine Monophosphate–Adenosine Monophosphate Synthase (cGAS), a Multifaceted Platform of Intracellular DNA Sensing

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