{"gene":"NCR3LG1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2009,"finding":"B7-H6 (NCR3LG1) is a tumor cell surface ligand that directly binds the activating NK cell receptor NKp30, triggering NKp30-mediated NK cell cytotoxicity and cytokine secretion. B7-H6 was not detected in normal human tissues but was expressed on human tumor cells.","method":"Protein identification, binding assays, NK cell activation assays","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — original identification with binding and functional activation assays; replicated extensively across multiple labs","pmids":["19528259"],"is_preprint":false},{"year":2011,"finding":"Crystal structure of the NKp30–B7-H6 complex was determined. NKp30 uses both front and back β-sheets of its Ig-like domain to engage B7-H6 via the side and face of the β-sandwich; B7-H6 contacts NKp30 through CDR-like loops of its V-like domain in an antibody-like interaction distinct from CTLA-4–B7 and PD-1–PD-L inhibitory complexes.","method":"X-ray crystallography of NKp30–B7-H6 complex","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 1 / Strong — high-resolution crystal structure with detailed structural validation; foundational mechanistic study","pmids":["21422170"],"is_preprint":false},{"year":2014,"finding":"Tumor cell shedding of B7-H6 ectodomain is mediated by the cell surface metalloproteases ADAM10 and ADAM17. Pharmacologic inhibition or siRNA knockdown of ADAM10/ADAM17 increased membrane B7-H6 levels and enhanced NKp30-mediated NK cell activation; soluble B7-H6 was elevated in sera of melanoma patients.","method":"Pharmacologic inhibitors of ADAMs, siRNA-mediated gene attenuation, flow cytometry, NK cell activation assays, patient serum ELISA","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — two orthogonal methods (pharmacologic + siRNA) identifying ADAM10 and ADAM17 as the sheddases; replicated across multiple cancer entities","pmids":["24780758"],"is_preprint":false},{"year":2013,"finding":"B7-H6 expression is induced on the surface of CD14+CD16+ proinflammatory monocytes and neutrophils upon TLR ligand stimulation or proinflammatory cytokines (IL-1β, TNF-α). A soluble form of B7-H6 is also produced by activated monocytes and neutrophils and is associated with membrane vesicles co-sedimenting with the exosomal fraction.","method":"In vitro stimulation assays, flow cytometry, fractionation/centrifugation of extracellular vesicles, in vivo patient cohort analysis","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (flow cytometry, vesicle fractionation, in vivo validation) in a single rigorous study","pmids":["23687088"],"is_preprint":false},{"year":2013,"finding":"B7-H6 mRNA and surface protein expression in tumor cells is downregulated by pan- or class I HDAC inhibitors (HDACi), and by siRNA-mediated knockdown of HDAC2 or HDAC3. This downregulation is associated with decreased B7-H6 reporter activity and reduced histone acetylation at the B7-H6 promoter, leading to impaired NKp30-dependent NK cell effector functions.","method":"Monoclonal antibody generation, siRNA knockdown of HDAC2/3, luciferase reporter assay, chromatin immunoprecipitation (ChIP), flow cytometry, NK functional assays","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (reporter assay, ChIP, siRNA, functional NK assay) in a single rigorous study","pmids":["23801635"],"is_preprint":false},{"year":2015,"finding":"Chronic engagement of NKp30 by soluble B7-H6 or tumor cell surface B7-H6 induces downregulation of NKp30 expression on NK cells, impairing NK cell IFN-γ production and cytolytic function against B7-H6-expressing target cells in ovarian carcinoma patients.","method":"Flow cytometry of tumor-associated NK cells from peritoneal fluids, in vitro functional assays with NK cells against B7-H6+ targets","journal":"Oncoimmunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — patient-derived cells with functional readouts; single lab but two orthogonal approaches (surface phenotyping + functional assay)","pmids":["26137398"],"is_preprint":false},{"year":2015,"finding":"Soluble B7-H6 in serum of high-risk neuroblastoma patients inhibits NK cell functions in vitro, and its serum concentration correlates with downregulation of NKp30 on circulating NK cells. NB cells expressing B7-H6 stimulate NK cells in an NKp30-dependent manner.","method":"In vitro NK cell functional assays with patient serum, flow cytometry, patient cohort analysis","journal":"Science translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional inhibition assays with patient-derived soluble B7-H6; single lab with multiple readouts","pmids":["25877893"],"is_preprint":false},{"year":2015,"finding":"Standard tumor therapeutics including chemotherapy (cisplatin, 5-FU), radiation, non-lethal heat shock, and TNF-α upregulate B7-H6 expression on tumor cells, enhancing their susceptibility to NK cell lysis. B7-H6 shRNA knockdown dampened this sensitization, establishing B7-H6 as the mediator of therapy-induced NK sensitivity.","method":"Drug/radiation treatment of tumor cell lines, shRNA knockdown, NK cell cytotoxicity assays, flow cytometry","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — shRNA rescue experiments with multiple therapeutic stimuli; single lab with genetic and pharmacologic orthogonal methods","pmids":["26472927"],"is_preprint":false},{"year":2016,"finding":"The proto-oncogene c-Myc and N-Myc directly drive B7-H6 expression by binding a functional site in the B7-H6 promoter. Pharmacological inhibition or siRNA/shRNA-mediated knockdown of c-Myc or N-Myc significantly decreased B7-H6 expression in melanoma, pancreatic carcinoma, and neuroblastoma cell lines, and impaired NKp30-mediated NK cell degranulation.","method":"Luciferase reporter assays, chromatin immunoprecipitation (ChIP), siRNA/shRNA knockdown, pharmacologic Myc inhibition, NK cell degranulation assays","journal":"Oncoimmunology","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — ChIP mapping of functional Myc binding site plus reporter assays and functional NK readout; multiple orthogonal methods, single lab","pmids":["27622013"],"is_preprint":false},{"year":2016,"finding":"Crystal structure of anti-B7-H6 antibody 17B1.3 in complex with B7-H6 ectodomain (2.5 Å) revealed that 17B1.3 binds a site on B7-H6 completely distinct from the NKp30-binding site. Structure-based mutations designed to disrupt potential B7-H6 dimerization through this site did not diminish NKp30-mediated cell activation, indicating that the antibody inhibits NK cell activation by sterically interfering with cell-cell contact rather than blocking the NKp30–B7-H6 interaction.","method":"X-ray crystallography (2.5 Å), site-directed mutagenesis, NK cell activation assays","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure plus mutagenesis plus functional assay in one study; single lab with multiple orthogonal methods","pmids":["27663271"],"is_preprint":false},{"year":2017,"finding":"Human cytomegalovirus (HCMV) upregulates B7-H6 expression upon infection, but the viral genes US18 and US20 counteract this by targeting B7-H6 for endosomal degradation, thereby reducing B7-H6 surface expression to evade NK cell recognition via NKp30.","method":"HCMV infection of cells, viral gene expression/deletion, flow cytometry, endosomal degradation pathway analysis","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic identification of specific viral genes (US18 and US20) with mechanistic follow-up showing endosomal degradation; single lab","pmids":["28819195"],"is_preprint":false},{"year":2019,"finding":"B7-H6 is expressed on the surface of HIV-2-infected CD4+ T cells, and chronic engagement of NKp30 by this surface B7-H6 induces downregulation of NKp30 on NK cells, impairing cytolytic function while maintaining IFN-γ overproduction, constituting a novel HIV-2 immune escape mechanism.","method":"Flow cytometry of NK cell phenotype and ligand expression in patient cohorts, functional NK cell assays against B7-H6+ target cells","journal":"AIDS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — patient cohort plus in vitro functional assays establishing the mechanism; single lab, multiple readouts","pmids":["30325780"],"is_preprint":false},{"year":2020,"finding":"B7-H6 promotes NHL cell proliferation, migration, and invasion via the Ras/MEK/ERK signaling pathway. B7-H6 knockdown reduced phosphorylation of Ras/MEK/ERK components, and the MEK inhibitor AZD8330 partially reversed proliferation and completely reversed migration/invasion induced by B7-H6 overexpression.","method":"siRNA/shRNA knockdown, quantitative phosphoproteomics, Western blot, B7-H6 overexpression, MEK inhibitor treatment, in vivo xenograft model, CCK-8 and Transwell assays","journal":"OncoTargets and therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phosphoproteomics plus genetic and pharmacologic validation of the Ras/MEK/ERK pathway; single lab, multiple orthogonal methods","pmids":["32606790"],"is_preprint":false},{"year":2021,"finding":"Soluble B7-H6 inhibits NK cell-mediated target cell killing and is associated with diminished cell surface expression of NKG2D and NCRs (NKp30, NKp40, NKp46). B7-H6 is only marginally released in association with extracellular vesicles (EVs), and EVs from B7-H6-expressing cells do not stimulate NK cell-mediated killing.","method":"Recombinant protein production in HEK293 cells, NK cell killing assays, flow cytometry, nano flow cytometry (NanoFCM) of EVs, ultracentrifugation fractionation","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (functional assays, EV fractionation, NanoFCM) in single lab study","pmids":["33671836"],"is_preprint":false},{"year":2021,"finding":"BRD4 acts as an epigenetic activator of B7-H6 transcription in AML cells. Histone acetylation mediated by CBP30/P300 facilitates BRD4 binding to the B7-H6 promoter, which recruits P-TEFb elongation factor to phosphorylate RNA polymerase II and activate B7-H6 transcription. BRD4 also co-binds with JMJD6 at a distal enhancer of the B7-H6 gene. Metformin modifies acetylation at the B7-H6 promoter, impairing BRD4 binding.","method":"BRD4 inhibition, ChIP, reporter assays, siRNA knockdown, metformin treatment, Western blot, flow cytometry, apoptosis assays","journal":"Oncoimmunology","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — ChIP mapping of BRD4 binding at B7-H6 promoter and enhancer plus functional validation with inhibitors and siRNA; single lab with multiple orthogonal methods","pmids":["33796404"],"is_preprint":false},{"year":2021,"finding":"All-trans retinoic acid (atRA) induces leukemia cell resistance to NK cell cytotoxicity by downregulating B7-H6 expression via reduction of c-Myc, which in turn inhibits B7-H6 transcription. DNA methylation and HDAC inhibitors could not restore B7-H6 mRNA, indicating epigenetic modifications are not responsible; the effect was specifically dependent on c-Myc signaling.","method":"CRISPR/Cas9 B7-H6 knockout, siRNA c-Myc knockdown, real-time PCR, flow cytometry, Western blotting, NK cell cytotoxicity assays","journal":"Cancer communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockouts/knockdowns plus pharmacologic epistasis ruling out alternative mechanisms; single lab, multiple orthogonal approaches","pmids":["34236140"],"is_preprint":false},{"year":2023,"finding":"B7-H6 contains 6 functional N-linked glycosylation sites. N43 and N208 glycosylation are conserved in jawed vertebrates and are essential for NK cell activation by B7-H6. Mechanistically, N208 glycosylation contributes to membrane stability and prevents B7-H6 shedding, while N43 glycosylation is required for B7-H6/NKp30 binding affinity.","method":"PNGase F digestion, tunicamycin blockade, single-site mutagenesis, SDS-PAGE, flow cytometry, NK cell activation assays, phylogenetic and structural analysis","journal":"Frontiers in immunology","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — site-directed mutagenesis of each glycosylation site plus functional assays establishing distinct mechanistic roles for N43 vs N208; single lab with multiple orthogonal methods","pmids":["38035117"],"is_preprint":false},{"year":2025,"finding":"SNRPA promotes HCC cell proliferation and lenvatinib resistance via B7-H6 by facilitating B7-H6 pre-mRNA maturation: SNRPA binds B7-H6 pre-mRNA directly and contributes to its intron 2 splicing. B7-H6 in turn activates STAT3/AKT signaling, promoting G1-S cell cycle transition and inhibiting apoptosis.","method":"Nanopore full-length cDNA sequencing, RNA-binding protein immunoprecipitation (RIP-seq), exon-exon/exon-intron junction PCR, siRNA/overexpression gain/loss-of-function, Western blot, flow cytometry, in vivo xenograft model","journal":"Bioscience trends","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RIP-seq plus splice-junction PCR plus gain/loss-of-function experiments; single lab, multiple orthogonal methods","pmids":["40240166"],"is_preprint":false},{"year":2025,"finding":"G-CSF derived from gastric cancer tumor microenvironment induces B7-H6 expression on tumor-infiltrating neutrophils via NF-κB signaling pathway activation. Blockade of B7-H6 on neutrophils promotes their apoptosis and shortens their lifespan.","method":"Flow cytometry, in vitro neutrophil stimulation with recombinant G-CSF and tumor supernatants, NF-κB pathway inhibition, apoptosis assays","journal":"Neoplasia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro stimulation with mechanistic pathway identification (NF-κB) plus functional blockade assay; single lab, multiple readouts","pmids":["40054066"],"is_preprint":false},{"year":2024,"finding":"CRISPR knockout of B7-H6 in CD4+ T cells reduced NK cell responses to HIV-1-infected cells in some donors, establishing that B7-H6 on HIV-infected cells contributes to NKp30-mediated NK recognition of HIV-infected cells.","method":"CRISPR editing of CD4+ T cells to knockout B7-H6, NK cell activation/response assays, ligand expression profiling by flow cytometry","journal":"bioRxiv (preprint)","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — CRISPR genetic knockout with functional NK readout; single study, preprint, noted donor variability","pmids":["bio_10.1101_2024.06.24.600449"],"is_preprint":true},{"year":2015,"finding":"NKp30-B7-H6 interaction in HBV-related acute-on-chronic liver failure (ACLF) triggers NK cell upregulation of IL-32, which in turn induces apoptosis of hepatocytes (Huh7 cells) in a dose-dependent manner. In vitro NK-92 cells killed high-B7-H6-expressing Huh7 and B7-H6-transfected LO2 hepatocytes in an NKp30/B7-H6-dependent manner.","method":"Co-culture assays, CCK8 cytotoxicity assay, anti-NKp30 antibody stimulation, Annexin V/PI apoptosis assay, flow cytometry, immunochemistry in patient samples","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic in vitro assays with antibody blockade and transfection establishing NKp30-B7-H6-IL-32 axis; single lab, multiple readouts","pmids":["26241657"],"is_preprint":false},{"year":2020,"finding":"B7-H6 is preferentially expressed in glioma stem-like cells (GSLCs) compared to parental glioma cell lines. Knockdown of B7-H6 by siRNA in GSLCs inhibited cell proliferation with decreased c-Myc expression and inactivation of PI3K/Akt and ERK/MAPK signaling pathways. RNMT (RNA guanine-7 methyltransferase) was identified as a downstream target co-expressed with B7-H6 and c-Myc; RNMT expression was inhibited upon B7-H6 knockdown. CRISPR-Cas9 knockout of B7-H6 in 293T cells also suppressed proliferation.","method":"siRNA knockdown, CRISPR-Cas9 knockout, Western blot, TCGA co-expression analysis, flow cytometry, proliferation assays","journal":"Journal of immunology research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — two genetic perturbation methods (siRNA + CRISPR) with pathway readouts; single lab","pmids":["32322592"],"is_preprint":false},{"year":2018,"finding":"B7-H6 knockdown in glioma cell lines (HepG2 and SMMC-7721) decreased expression of C-myc, C-fos, and Cyclin-D1, and was accompanied by reduced cell proliferation, migration, invasion, and G1 cell cycle arrest, indicating B7-H6 regulates these oncogenic factors intracellularly.","method":"RNA interference (siRNA), CCK-8 proliferation assay, migration/invasion assays, Western blot, flow cytometry cell cycle analysis","journal":"Cancer cell international","confidence":"Low","confidence_rationale":"Tier 3 / Weak — siRNA knockdown with downstream expression readouts; single lab, no pathway rescue experiments","pmids":["30186042"],"is_preprint":false},{"year":2018,"finding":"Lipopolysaccharide (LPS) induces B7-H6 expression in glioma cell lines. B7-H6 knockdown increased E-cadherin and Bax expression while decreasing vimentin, N-cadherin, MMP-2, MMP-9, and survivin, suggesting B7-H6 promotes EMT and invasion-related signaling.","method":"LPS stimulation, siRNA knockdown, Western blot, cell viability/migration/invasion assays","journal":"International immunopharmacology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single siRNA knockdown with downstream protein expression readouts; single lab, no mechanistic pathway rescue","pmids":["29679856"],"is_preprint":false},{"year":2023,"finding":"B7-H6 promotes migration and invasion of medulloblastoma (Daoy) cells by activating c-MYC, which in turn enhances F-actin cytoskeleton rearrangement and upregulates MMP-2 and MMP-9 expression.","method":"siRNA knockdown, plasmid overexpression, Transwell migration/invasion assays, Western blot, immunofluorescence staining of F-actin","journal":"Medical oncology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, siRNA/overexpression with functional readouts but no pathway rescue or genetic epistasis experiments","pmids":["36692844"],"is_preprint":false}],"current_model":"B7-H6 (NCR3LG1) is a B7-family transmembrane ligand whose extracellular V-like domain binds the activating NK receptor NKp30 (crystal structure solved at 2.5 Å) to trigger NK cell cytotoxicity and cytokine secretion; it is absent from normal tissues but induced on tumor cells and inflammatory monocytes/neutrophils (via TLR/cytokine and NF-κB pathways), regulated transcriptionally by c-Myc/N-Myc and the epigenetic reader BRD4 (via CBP/P300 acetylation), cleaved from the cell surface by ADAM10/ADAM17 metalloproteases to generate soluble B7-H6 that downregulates NKp30 on NK cells, requires N-linked glycosylation at N43 (for NKp30 binding) and N208 (for membrane stability) for full function, and is exploited by HCMV (via US18/US20-mediated endosomal degradation) and HIV-2 to evade NK surveillance; intracellularly, B7-H6 activates Ras/MEK/ERK, PI3K/Akt, and STAT3/AKT signaling to promote tumor cell proliferation, migration, and survival."},"narrative":{"mechanistic_narrative":"B7-H6 (NCR3LG1) is a B7-family cell-surface ligand that links cellular stress and malignant transformation to innate immune surveillance by directly engaging the activating NK receptor NKp30 to trigger NK cell cytotoxicity and cytokine secretion [PMID:19528259]. The structural basis of this interaction is an antibody-like engagement in which NKp30 uses both faces of its Ig-like domain to contact CDR-like loops of the B7-H6 V-like domain, a geometry distinct from the inhibitory CTLA-4–B7 and PD-1–PD-L complexes [PMID:21422170]. B7-H6 is largely absent from normal tissue and is induced on tumor cells and on proinflammatory CD14+CD16+ monocytes and neutrophils following TLR ligation or proinflammatory cytokines, including a G-CSF/NF-κB axis on tumor-infiltrating neutrophils [PMID:19528259, PMID:23687088, PMID:40054066]. Its expression is transcriptionally driven by c-Myc and N-Myc binding the promoter and by an epigenetic program in which CBP/P300-dependent histone acetylation recruits BRD4 to promoter and enhancer elements [PMID:27622013, PMID:33796404], and it is further upregulated by chemotherapy, radiation, and heat shock to sensitize tumor cells to NK lysis [PMID:26472927]. Surface B7-H6 is shed by the metalloproteases ADAM10 and ADAM17 to generate soluble ligand that, through chronic NKp30 engagement, downregulates NKp30 and impairs NK cell function—a route exploited as immune escape in cancer and HIV-2 infection [PMID:24780758, PMID:26137398, PMID:30325780, PMID:33671836]. N-linked glycosylation governs function site-specifically: N43 glycosylation is required for NKp30-binding affinity, while N208 glycosylation stabilizes the protein at the membrane and prevents shedding [PMID:38035117]. Beyond its role as an immune ligand, B7-H6 acts intracellularly to promote tumor cell proliferation, migration, and invasion through Ras/MEK/ERK, PI3K/Akt, and STAT3/AKT signaling, with c-Myc as a recurrent downstream effector [PMID:32606790, PMID:40240166, PMID:32322592]. Viral antagonism is also documented: HCMV genes US18 and US20 target B7-H6 for endosomal degradation to evade NKp30-mediated recognition [PMID:28819195].","teleology":[{"year":2009,"claim":"Established the existence and function of B7-H6, answering whether a dedicated tumor-restricted ligand drives NKp30 activation.","evidence":"Protein identification with binding and NK activation assays on human tumor cells","pmids":["19528259"],"confidence":"High","gaps":["Did not resolve the structural basis of recognition","Regulation of its tumor-restricted expression unknown"]},{"year":2011,"claim":"Defined the structural basis of NKp30–B7-H6 recognition, showing an antibody-like engagement distinct from inhibitory B7-family complexes.","evidence":"X-ray crystallography of the NKp30–B7-H6 complex","pmids":["21422170"],"confidence":"High","gaps":["Did not address post-translational requirements for binding","Affinity modulation in vivo not resolved"]},{"year":2013,"claim":"Identified how B7-H6 expression is induced, linking it both to inflammatory cell activation and to chromatin-level transcriptional control.","evidence":"TLR/cytokine stimulation of monocytes/neutrophils with vesicle fractionation; HDAC2/3 knockdown with promoter ChIP and reporter assays","pmids":["23687088","23801635"],"confidence":"High","gaps":["Direct transcription factors not yet identified","Relationship between vesicle-associated and shed soluble forms unclear"]},{"year":2014,"claim":"Showed that surface B7-H6 is proteolytically shed, explaining the origin of soluble ligand and a mechanism limiting NK activation.","evidence":"ADAM10/ADAM17 pharmacologic inhibition and siRNA with flow cytometry, NK assays, and patient serum ELISA","pmids":["24780758"],"confidence":"High","gaps":["Did not establish functional consequences of soluble form on NK receptors","Cleavage site not mapped"]},{"year":2015,"claim":"Demonstrated that chronic NKp30 engagement by soluble or surface B7-H6 downregulates NKp30 and other NK receptors, defining a self-limiting immune-escape circuit.","evidence":"Patient-derived NK cell phenotyping and functional assays across ovarian carcinoma, neuroblastoma, and HBV-related liver failure cohorts","pmids":["26137398","25877893","26241657"],"confidence":"Medium","gaps":["Molecular mechanism of NKp30 downregulation not defined","Reversibility of receptor suppression untested"]},{"year":2015,"claim":"Established B7-H6 as the mediator of therapy-induced NK sensitivity, connecting cellular stress to immune recognition.","evidence":"Chemotherapy/radiation/heat-shock treatment with B7-H6 shRNA knockdown and NK cytotoxicity assays","pmids":["26472927"],"confidence":"Medium","gaps":["Stress-responsive transcriptional sensors not identified","Single-lab finding"]},{"year":2016,"claim":"Identified c-Myc/N-Myc as direct transcriptional drivers of B7-H6 and showed that anti-B7-H6 antibodies can block activation sterically rather than competitively.","evidence":"Promoter ChIP, reporter assays, Myc knockdown/inhibition; crystallography of 17B1.3–B7-H6 with mutagenesis","pmids":["27622013","27663271"],"confidence":"High","gaps":["Whether B7-H6 dimerizes physiologically remained unresolved","Myc cofactors at the promoter not mapped"]},{"year":2017,"claim":"Revealed viral antagonism of B7-H6, showing HCMV degrades the ligand to evade NK surveillance.","evidence":"HCMV infection with US18/US20 gene deletion, flow cytometry, endosomal degradation analysis","pmids":["28819195"],"confidence":"Medium","gaps":["Molecular details of US18/US20-mediated targeting not fully resolved","Single lab"]},{"year":2019,"claim":"Extended the immune-escape paradigm to retroviral infection, showing HIV-2-infected T cells express B7-H6 to chronically suppress NKp30.","evidence":"Patient cohort flow cytometry and NK functional assays against B7-H6+ targets","pmids":["30325780"],"confidence":"Medium","gaps":["Why HIV-2 induces rather than degrades B7-H6 not explained","Selective loss of cytolysis but not IFN-γ unexplained"]},{"year":2021,"claim":"Defined the epigenetic activation program and intracellular oncogenic signaling of B7-H6, beyond its ligand role.","evidence":"BRD4/CBP-P300 ChIP and inhibitor studies in AML; Ras/MEK/ERK phosphoproteomics and inhibitor rescue in lymphoma; atRA/c-Myc epistasis in leukemia","pmids":["33796404","32606790","34236140"],"confidence":"High","gaps":["How a surface ligand drives intracellular kinase signaling mechanistically unclear","Receptor/effector linking B7-H6 to Ras/MEK/ERK not identified"]},{"year":2023,"claim":"Resolved site-specific roles of N-glycosylation, separating ligand-binding competence (N43) from membrane stability and shedding resistance (N208).","evidence":"Single-site mutagenesis, PNGase F/tunicamycin treatment, NK activation assays, phylogenetic analysis","pmids":["38035117"],"confidence":"High","gaps":["Roles of the other four glycosylation sites undefined","Glycan structures themselves not characterized"]},{"year":2025,"claim":"Connected B7-H6 expression to RNA processing and downstream STAT3/AKT-driven proliferation, broadening its tumor-intrinsic functions.","evidence":"SNRPA RIP-seq and splice-junction PCR with gain/loss-of-function in HCC; G-CSF/NF-κB induction in neutrophils with apoptosis assays","pmids":["40240166","40054066"],"confidence":"Medium","gaps":["Direct receptor mediating intracellular signaling unknown","Generalizability across tumor types untested"]},{"year":null,"claim":"How a cell-surface ligand transduces the intracellular Ras/MEK/ERK, PI3K/Akt, and STAT3/AKT signals attributed to B7-H6 remains the central unresolved mechanistic question.","evidence":"No discovery in the corpus identifies a cytoplasmic effector or receptor coupling B7-H6 to these pathways","pmids":[],"confidence":"Medium","gaps":["No defined intracellular signaling partner","Distinction between cell-autonomous and NKp30-engagement effects unresolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[0,1]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,2,16]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[2,3,13]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[0,1,5]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[12,17,21]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[8,14]}],"complexes":[],"partners":["NKP30","ADAM10","ADAM17"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q68D85","full_name":"Natural cytotoxicity triggering receptor 3 ligand 1","aliases":["B7 homolog 6","B7-H6"],"length_aa":454,"mass_kda":50.8,"function":"Triggers NCR3-dependent natural killer cell activation","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q68D85/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NCR3LG1","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"GGPS1","stoichiometry":0.2},{"gene":"PSMB3","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/NCR3LG1","total_profiled":1310},"omim":[{"mim_id":"613714","title":"NATURAL KILLER CELL CYTOTOXICITY RECEPTOR 3 LIGAND 1; NCR3LG1","url":"https://www.omim.org/entry/613714"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"brain","ntpm":6.0},{"tissue":"parathyroid gland","ntpm":7.0}],"url":"https://www.proteinatlas.org/search/NCR3LG1"},"hgnc":{"alias_symbol":["DKFZp686O24166","B7-H6"],"prev_symbol":[]},"alphafold":{"accession":"Q68D85","domains":[{"cath_id":"2.60.40.10","chopping":"27-141","consensus_level":"high","plddt":94.725,"start":27,"end":141},{"cath_id":"2.60.40.10","chopping":"145-246","consensus_level":"high","plddt":89.1385,"start":145,"end":246},{"cath_id":"1.10.150.180","chopping":"298-384","consensus_level":"high","plddt":88.6775,"start":298,"end":384}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q68D85","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q68D85-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q68D85-F1-predicted_aligned_error_v6.png","plddt_mean":77.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NCR3LG1","jax_strain_url":"https://www.jax.org/strain/search?query=NCR3LG1"},"sequence":{"accession":"Q68D85","fasta_url":"https://rest.uniprot.org/uniprotkb/Q68D85.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q68D85/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q68D85"}},"corpus_meta":[{"pmid":"19528259","id":"PMC_19528259","title":"The B7 family member B7-H6 is a tumor cell ligand for the activating natural killer cell receptor NKp30 in humans.","date":"2009","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/19528259","citation_count":548,"is_preprint":false},{"pmid":"24780758","id":"PMC_24780758","title":"Metalloprotease-mediated tumor cell shedding of B7-H6, the ligand of the natural killer cell-activating receptor NKp30.","date":"2014","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/24780758","citation_count":172,"is_preprint":false},{"pmid":"26137398","id":"PMC_26137398","title":"B7-H6-mediated downregulation of NKp30 in NK cells contributes to ovarian carcinoma immune escape.","date":"2015","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/26137398","citation_count":153,"is_preprint":false},{"pmid":"25877893","id":"PMC_25877893","title":"Clinical impact of the NKp30/B7-H6 axis in high-risk neuroblastoma patients.","date":"2015","source":"Science translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/25877893","citation_count":124,"is_preprint":false},{"pmid":"23687088","id":"PMC_23687088","title":"Induction of B7-H6, a ligand for the natural killer cell-activating receptor NKp30, in inflammatory conditions.","date":"2013","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/23687088","citation_count":120,"is_preprint":false},{"pmid":"23801635","id":"PMC_23801635","title":"Downregulation of the activating NKp30 ligand B7-H6 by HDAC inhibitors impairs tumor cell recognition by NK cells.","date":"2013","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/23801635","citation_count":114,"is_preprint":false},{"pmid":"21422170","id":"PMC_21422170","title":"Structure of the human activating natural cytotoxicity receptor NKp30 bound to its tumor cell ligand B7-H6.","date":"2011","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/21422170","citation_count":100,"is_preprint":false},{"pmid":"21877119","id":"PMC_21877119","title":"B7-H6/NKp30 interaction: a mechanism of alerting NK cells against tumors.","date":"2011","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/21877119","citation_count":90,"is_preprint":false},{"pmid":"26472927","id":"PMC_26472927","title":"Tumor Therapeutics Work as Stress Inducers to Enhance Tumor Sensitivity to Natural Killer (NK) Cell Cytolysis by Up-regulating NKp30 Ligand B7-H6.","date":"2015","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/26472927","citation_count":79,"is_preprint":false},{"pmid":"26464699","id":"PMC_26464699","title":"B7-H6 expression correlates with cancer progression and patient's survival in human ovarian cancer.","date":"2015","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26464699","citation_count":64,"is_preprint":false},{"pmid":"27622013","id":"PMC_27622013","title":"The proto-oncogene Myc drives expression of the NK cell-activating NKp30 ligand B7-H6 in tumor cells.","date":"2016","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/27622013","citation_count":48,"is_preprint":false},{"pmid":"23066150","id":"PMC_23066150","title":"Mimicking an induced self phenotype by coating lymphomas with the NKp30 ligand B7-H6 promotes NK cell cytotoxicity.","date":"2012","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/23066150","citation_count":47,"is_preprint":false},{"pmid":"26891663","id":"PMC_26891663","title":"Knockdown of B7-H6 inhibits tumor progression and enhances chemosensitivity in B-cell non-Hodgkin lymphoma.","date":"2016","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/26891663","citation_count":41,"is_preprint":false},{"pmid":"33268483","id":"PMC_33268483","title":"Affinity Maturation of B7-H6 Translates into Enhanced NK Cell-Mediated Tumor Cell Lysis and Improved Proinflammatory Cytokine Release of Bispecific Immunoligands via NKp30 Engagement.","date":"2020","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/33268483","citation_count":40,"is_preprint":false},{"pmid":"28819195","id":"PMC_28819195","title":"Human cytomegalovirus escapes immune recognition by NK cells through the downregulation of B7-H6 by the viral genes US18 and US20.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28819195","citation_count":40,"is_preprint":false},{"pmid":"25400778","id":"PMC_25400778","title":"B7-H6 expression in non-small cell lung cancers.","date":"2014","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/25400778","citation_count":39,"is_preprint":false},{"pmid":"28415577","id":"PMC_28415577","title":"High expression of B7-H6 in human glioma tissues promotes tumor progression.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/28415577","citation_count":39,"is_preprint":false},{"pmid":"29679856","id":"PMC_29679856","title":"B7-H6 expression is induced by lipopolysaccharide and facilitates cancer invasion and metastasis in human gliomas.","date":"2018","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/29679856","citation_count":36,"is_preprint":false},{"pmid":"29086181","id":"PMC_29086181","title":"The B7 Family Member B7-H6: a New Bane of Tumor.","date":"2017","source":"Pathology oncology research : POR","url":"https://pubmed.ncbi.nlm.nih.gov/29086181","citation_count":35,"is_preprint":false},{"pmid":"33161972","id":"PMC_33161972","title":"Anti-fouling SERS-based immunosensor for point-of-care detection of the B7-H6 tumor biomarker in cervical cancer patient serum.","date":"2020","source":"Analytica chimica acta","url":"https://pubmed.ncbi.nlm.nih.gov/33161972","citation_count":35,"is_preprint":false},{"pmid":"30094100","id":"PMC_30094100","title":"Long non-coding RNA LINC00673 promotes breast cancer proliferation and metastasis through regulating B7-H6 and epithelial-mesenchymal transition.","date":"2018","source":"American journal of cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/30094100","citation_count":34,"is_preprint":false},{"pmid":"32322592","id":"PMC_32322592","title":"Preferential Expression of B7-H6 in Glioma Stem-Like Cells Enhances Tumor Cell Proliferation via the c-Myc/RNMT Axis.","date":"2020","source":"Journal of immunology research","url":"https://pubmed.ncbi.nlm.nih.gov/32322592","citation_count":30,"is_preprint":false},{"pmid":"33839362","id":"PMC_33839362","title":"Beyond Programmed Death-Ligand 1: B7-H6 Emerges as a Potential Immunotherapy Target in SCLC.","date":"2021","source":"Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/33839362","citation_count":27,"is_preprint":false},{"pmid":"28789456","id":"PMC_28789456","title":"Clinical significance of novel costimulatory molecule B7-H6 in human breast cancer.","date":"2017","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/28789456","citation_count":24,"is_preprint":false},{"pmid":"33671836","id":"PMC_33671836","title":"Secreted Ligands of the NK Cell Receptor NKp30: B7-H6 Is in Contrast to BAG6 Only Marginally Released via Extracellular Vesicles.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33671836","citation_count":23,"is_preprint":false},{"pmid":"31904350","id":"PMC_31904350","title":"Immunological role and underlying mechanisms of B7-H6 in tumorigenesis.","date":"2020","source":"Clinica chimica acta; international journal of clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31904350","citation_count":22,"is_preprint":false},{"pmid":"35697295","id":"PMC_35697295","title":"The potential of B7-H6 as a therapeutic target in cancer immunotherapy.","date":"2022","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35697295","citation_count":21,"is_preprint":false},{"pmid":"30186042","id":"PMC_30186042","title":"B7-H6 expression in human hepatocellular carcinoma and its clinical significance [corrected].","date":"2018","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/30186042","citation_count":19,"is_preprint":false},{"pmid":"36166004","id":"PMC_36166004","title":"A Novel B7-H6-Targeted IgG-Like T Cell-Engaging Antibody for the Treatment of Gastrointestinal Tumors.","date":"2022","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/36166004","citation_count":18,"is_preprint":false},{"pmid":"32606790","id":"PMC_32606790","title":"B7-H6 Promotes Cell Proliferation, Migration and Invasion of Non-Hodgkin Lymphoma via Ras/MEK/ERK Pathway Based on Quantitative Phosphoproteomics Data.","date":"2020","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/32606790","citation_count":16,"is_preprint":false},{"pmid":"33172426","id":"PMC_33172426","title":"B7-H6, an immunoligand for the natural killer cell activating receptor NKp30, reveals inhibitory effects on cell proliferation and migration, but not apoptosis, in cervical cancer derived-cell lines.","date":"2020","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/33172426","citation_count":16,"is_preprint":false},{"pmid":"26241657","id":"PMC_26241657","title":"NKP30-B7-H6 Interaction Aggravates Hepatocyte Damage through Up-Regulation of Interleukin-32 Expression in Hepatitis B Virus-Related Acute-On-Chronic Liver Failure.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26241657","citation_count":15,"is_preprint":false},{"pmid":"32138659","id":"PMC_32138659","title":"Positive staining of the immunoligand B7-H6 in abnormal/transformed keratinocytes consistently accompanies the progression of cervical cancer.","date":"2020","source":"BMC immunology","url":"https://pubmed.ncbi.nlm.nih.gov/32138659","citation_count":15,"is_preprint":false},{"pmid":"32770284","id":"PMC_32770284","title":"B7-H6 as an efficient target for T cell-induced cytotoxicity in haematologic malignant cells.","date":"2020","source":"Investigational new drugs","url":"https://pubmed.ncbi.nlm.nih.gov/32770284","citation_count":14,"is_preprint":false},{"pmid":"32566616","id":"PMC_32566616","title":"Expression of a novel immune checkpoint B7-H6 ligand in human small cell lung cancer.","date":"2020","source":"Annals of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/32566616","citation_count":14,"is_preprint":false},{"pmid":"30325780","id":"PMC_30325780","title":"B7-H6-mediated downregulation of NKp30 in natural killer cells contributes to HIV-2 immune escape.","date":"2019","source":"AIDS (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/30325780","citation_count":14,"is_preprint":false},{"pmid":"27663271","id":"PMC_27663271","title":"Structural Insights into the Inhibitory Mechanism of an Antibody against B7-H6, a Stress-Induced Cellular Ligand for the Natural Killer Cell Receptor NKp30.","date":"2016","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/27663271","citation_count":14,"is_preprint":false},{"pmid":"33824367","id":"PMC_33824367","title":"Immune checkpoint molecules B7-H6 and PD-L1 co-pattern the tumor inflammatory microenvironment in human breast cancer.","date":"2021","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33824367","citation_count":12,"is_preprint":false},{"pmid":"33796404","id":"PMC_33796404","title":"Bromodomain protein BRD4 is an epigenetic activator of B7-H6 expression in acute myeloid leukemia.","date":"2021","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/33796404","citation_count":11,"is_preprint":false},{"pmid":"33708955","id":"PMC_33708955","title":"B7-H6 is a new potential biomarker and therapeutic target of T-lymphoblastic lymphoma.","date":"2021","source":"Annals of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33708955","citation_count":11,"is_preprint":false},{"pmid":"39408655","id":"PMC_39408655","title":"Harnessing B7-H6 for Anticancer Immunotherapy: Expression, Pathways, and Therapeutic Strategies.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39408655","citation_count":8,"is_preprint":false},{"pmid":"34460046","id":"PMC_34460046","title":"Sevoflurane impedes glioma progression via regulating circ_0000215/miR-1200/NCR3LG1 axis.","date":"2021","source":"Metabolic brain disease","url":"https://pubmed.ncbi.nlm.nih.gov/34460046","citation_count":8,"is_preprint":false},{"pmid":"34236140","id":"PMC_34236140","title":"All-trans retinoic acid induces leukemia resistance to NK cell cytotoxicity by down-regulating B7-H6 expression via c-Myc signaling.","date":"2021","source":"Cancer communications (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/34236140","citation_count":8,"is_preprint":false},{"pmid":"26057798","id":"PMC_26057798","title":"Expression, crystallization and X-ray diffraction analysis of a complex between B7-H6, a tumor cell ligand for the natural cytotoxicity receptor NKp30, and an inhibitory antibody.","date":"2015","source":"Acta crystallographica. Section F, Structural biology communications","url":"https://pubmed.ncbi.nlm.nih.gov/26057798","citation_count":8,"is_preprint":false},{"pmid":"38035117","id":"PMC_38035117","title":"Deficiency of N-linked glycosylation impairs immune function of B7-H6.","date":"2023","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/38035117","citation_count":7,"is_preprint":false},{"pmid":"31933861","id":"PMC_31933861","title":"Expression of B7-H6 in chronic myeloid leukemia and its clinical significance.","date":"2019","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/31933861","citation_count":7,"is_preprint":false},{"pmid":"34280008","id":"PMC_34280008","title":"B7-H6 as a Diagnostic Biomarker for Cervical Squamous Cell Carcinoma.","date":"2021","source":"Genetic testing and molecular biomarkers","url":"https://pubmed.ncbi.nlm.nih.gov/34280008","citation_count":5,"is_preprint":false},{"pmid":"30214376","id":"PMC_30214376","title":"Correction to: Expression of B7-H6 expression in human hepatocellular carcinoma and its clinical significance.","date":"2018","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/30214376","citation_count":4,"is_preprint":false},{"pmid":"39811682","id":"PMC_39811682","title":"Enhancing NK cell-mediated tumor killing of B7-H6+ cells with bispecific antibodies targeting allosteric sites of NKp30.","date":"2024","source":"Molecular therapy. Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/39811682","citation_count":3,"is_preprint":false},{"pmid":"40240166","id":"PMC_40240166","title":"SNRPA promotes hepatocellular carcinoma proliferation and lenvatinib resistance via B7-H6-STAT3/AKT axis by facilitating B7-H6 pre-mRNA maturation.","date":"2025","source":"Bioscience trends","url":"https://pubmed.ncbi.nlm.nih.gov/40240166","citation_count":2,"is_preprint":false},{"pmid":"40873574","id":"PMC_40873574","title":"Dual T/NK cell engagement via B7-H6-targeted bispecific antibodies and IL-15 eradicates chemo-resistant solid tumors.","date":"2025","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/40873574","citation_count":2,"is_preprint":false},{"pmid":"36692844","id":"PMC_36692844","title":"B7-H6 enhances F-actin rearrangement by targeting c-MYC activation to promote medulloblastoma migration and invasion.","date":"2023","source":"Medical oncology (Northwood, London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/36692844","citation_count":2,"is_preprint":false},{"pmid":"40054066","id":"PMC_40054066","title":"Expression, regulation, function and clinical significance of B7-H6 on neutrophils in human gastric cancer.","date":"2025","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/40054066","citation_count":1,"is_preprint":false},{"pmid":"41001137","id":"PMC_41001137","title":"The B7 family molecules in oral squamous cell carcinoma: a systematic review. Part II: B7-1, B7-2, B7-H2, B7-H3, B7-H4, B7-H5 (VISTA), B7-H6 and B7-H7.","date":"2020","source":"Postepy dermatologii i alergologii","url":"https://pubmed.ncbi.nlm.nih.gov/41001137","citation_count":1,"is_preprint":false},{"pmid":"37405651","id":"PMC_37405651","title":"Affinity Maturation of the Natural Ligand (B7-H6) for Natural Cytotoxicity Receptor NKp30 by Yeast Surface Display.","date":"2023","source":"Methods in molecular biology (Clifton, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/37405651","citation_count":1,"is_preprint":false},{"pmid":"25001651","id":"PMC_25001651","title":"Allogeneic and xenogeneic anti-tumor effect of callithrix jacchus natural killer cells is dependent on NKp30 and B7-H6 interaction.","date":"2014","source":"Journal of biological regulators and homeostatic agents","url":"https://pubmed.ncbi.nlm.nih.gov/25001651","citation_count":1,"is_preprint":false},{"pmid":"27609569","id":"PMC_27609569","title":"[Elevated expression of B7-H6 in U87 cells-derived glioma stem like cells is associated with biological characteristics].","date":"2016","source":"Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/27609569","citation_count":1,"is_preprint":false},{"pmid":"40425991","id":"PMC_40425991","title":"Association between elevated serum soluble B7-H6 and infection in hemodialysis patients.","date":"2025","source":"Clinical and experimental nephrology","url":"https://pubmed.ncbi.nlm.nih.gov/40425991","citation_count":0,"is_preprint":false},{"pmid":"35261813","id":"PMC_35261813","title":"Erratum: Long non-coding RNA LINC00673 promotes breast cancer proliferation and metastasis through regulating B7-H6 and epithelial-mesenchymal transition.","date":"2022","source":"American journal of cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/35261813","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.06.24.600449","title":"NKp30 and NKG2D contribute to natural killer recognition of HIV-infected cells","date":"2024-06-27","source":"bioRxiv","url":"https://doi.org/10.1101/2024.06.24.600449","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":32382,"output_tokens":6599,"usd":0.098065,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":15477,"output_tokens":3892,"usd":0.087343,"stage2_stop_reason":"end_turn"},"total_usd":0.185408,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2009,\n      \"finding\": \"B7-H6 (NCR3LG1) is a tumor cell surface ligand that directly binds the activating NK cell receptor NKp30, triggering NKp30-mediated NK cell cytotoxicity and cytokine secretion. B7-H6 was not detected in normal human tissues but was expressed on human tumor cells.\",\n      \"method\": \"Protein identification, binding assays, NK cell activation assays\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — original identification with binding and functional activation assays; replicated extensively across multiple labs\",\n      \"pmids\": [\"19528259\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Crystal structure of the NKp30–B7-H6 complex was determined. NKp30 uses both front and back β-sheets of its Ig-like domain to engage B7-H6 via the side and face of the β-sandwich; B7-H6 contacts NKp30 through CDR-like loops of its V-like domain in an antibody-like interaction distinct from CTLA-4–B7 and PD-1–PD-L inhibitory complexes.\",\n      \"method\": \"X-ray crystallography of NKp30–B7-H6 complex\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — high-resolution crystal structure with detailed structural validation; foundational mechanistic study\",\n      \"pmids\": [\"21422170\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Tumor cell shedding of B7-H6 ectodomain is mediated by the cell surface metalloproteases ADAM10 and ADAM17. Pharmacologic inhibition or siRNA knockdown of ADAM10/ADAM17 increased membrane B7-H6 levels and enhanced NKp30-mediated NK cell activation; soluble B7-H6 was elevated in sera of melanoma patients.\",\n      \"method\": \"Pharmacologic inhibitors of ADAMs, siRNA-mediated gene attenuation, flow cytometry, NK cell activation assays, patient serum ELISA\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — two orthogonal methods (pharmacologic + siRNA) identifying ADAM10 and ADAM17 as the sheddases; replicated across multiple cancer entities\",\n      \"pmids\": [\"24780758\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"B7-H6 expression is induced on the surface of CD14+CD16+ proinflammatory monocytes and neutrophils upon TLR ligand stimulation or proinflammatory cytokines (IL-1β, TNF-α). A soluble form of B7-H6 is also produced by activated monocytes and neutrophils and is associated with membrane vesicles co-sedimenting with the exosomal fraction.\",\n      \"method\": \"In vitro stimulation assays, flow cytometry, fractionation/centrifugation of extracellular vesicles, in vivo patient cohort analysis\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (flow cytometry, vesicle fractionation, in vivo validation) in a single rigorous study\",\n      \"pmids\": [\"23687088\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"B7-H6 mRNA and surface protein expression in tumor cells is downregulated by pan- or class I HDAC inhibitors (HDACi), and by siRNA-mediated knockdown of HDAC2 or HDAC3. This downregulation is associated with decreased B7-H6 reporter activity and reduced histone acetylation at the B7-H6 promoter, leading to impaired NKp30-dependent NK cell effector functions.\",\n      \"method\": \"Monoclonal antibody generation, siRNA knockdown of HDAC2/3, luciferase reporter assay, chromatin immunoprecipitation (ChIP), flow cytometry, NK functional assays\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (reporter assay, ChIP, siRNA, functional NK assay) in a single rigorous study\",\n      \"pmids\": [\"23801635\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Chronic engagement of NKp30 by soluble B7-H6 or tumor cell surface B7-H6 induces downregulation of NKp30 expression on NK cells, impairing NK cell IFN-γ production and cytolytic function against B7-H6-expressing target cells in ovarian carcinoma patients.\",\n      \"method\": \"Flow cytometry of tumor-associated NK cells from peritoneal fluids, in vitro functional assays with NK cells against B7-H6+ targets\",\n      \"journal\": \"Oncoimmunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — patient-derived cells with functional readouts; single lab but two orthogonal approaches (surface phenotyping + functional assay)\",\n      \"pmids\": [\"26137398\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Soluble B7-H6 in serum of high-risk neuroblastoma patients inhibits NK cell functions in vitro, and its serum concentration correlates with downregulation of NKp30 on circulating NK cells. NB cells expressing B7-H6 stimulate NK cells in an NKp30-dependent manner.\",\n      \"method\": \"In vitro NK cell functional assays with patient serum, flow cytometry, patient cohort analysis\",\n      \"journal\": \"Science translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional inhibition assays with patient-derived soluble B7-H6; single lab with multiple readouts\",\n      \"pmids\": [\"25877893\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Standard tumor therapeutics including chemotherapy (cisplatin, 5-FU), radiation, non-lethal heat shock, and TNF-α upregulate B7-H6 expression on tumor cells, enhancing their susceptibility to NK cell lysis. B7-H6 shRNA knockdown dampened this sensitization, establishing B7-H6 as the mediator of therapy-induced NK sensitivity.\",\n      \"method\": \"Drug/radiation treatment of tumor cell lines, shRNA knockdown, NK cell cytotoxicity assays, flow cytometry\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — shRNA rescue experiments with multiple therapeutic stimuli; single lab with genetic and pharmacologic orthogonal methods\",\n      \"pmids\": [\"26472927\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The proto-oncogene c-Myc and N-Myc directly drive B7-H6 expression by binding a functional site in the B7-H6 promoter. Pharmacological inhibition or siRNA/shRNA-mediated knockdown of c-Myc or N-Myc significantly decreased B7-H6 expression in melanoma, pancreatic carcinoma, and neuroblastoma cell lines, and impaired NKp30-mediated NK cell degranulation.\",\n      \"method\": \"Luciferase reporter assays, chromatin immunoprecipitation (ChIP), siRNA/shRNA knockdown, pharmacologic Myc inhibition, NK cell degranulation assays\",\n      \"journal\": \"Oncoimmunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — ChIP mapping of functional Myc binding site plus reporter assays and functional NK readout; multiple orthogonal methods, single lab\",\n      \"pmids\": [\"27622013\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Crystal structure of anti-B7-H6 antibody 17B1.3 in complex with B7-H6 ectodomain (2.5 Å) revealed that 17B1.3 binds a site on B7-H6 completely distinct from the NKp30-binding site. Structure-based mutations designed to disrupt potential B7-H6 dimerization through this site did not diminish NKp30-mediated cell activation, indicating that the antibody inhibits NK cell activation by sterically interfering with cell-cell contact rather than blocking the NKp30–B7-H6 interaction.\",\n      \"method\": \"X-ray crystallography (2.5 Å), site-directed mutagenesis, NK cell activation assays\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure plus mutagenesis plus functional assay in one study; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"27663271\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Human cytomegalovirus (HCMV) upregulates B7-H6 expression upon infection, but the viral genes US18 and US20 counteract this by targeting B7-H6 for endosomal degradation, thereby reducing B7-H6 surface expression to evade NK cell recognition via NKp30.\",\n      \"method\": \"HCMV infection of cells, viral gene expression/deletion, flow cytometry, endosomal degradation pathway analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic identification of specific viral genes (US18 and US20) with mechanistic follow-up showing endosomal degradation; single lab\",\n      \"pmids\": [\"28819195\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"B7-H6 is expressed on the surface of HIV-2-infected CD4+ T cells, and chronic engagement of NKp30 by this surface B7-H6 induces downregulation of NKp30 on NK cells, impairing cytolytic function while maintaining IFN-γ overproduction, constituting a novel HIV-2 immune escape mechanism.\",\n      \"method\": \"Flow cytometry of NK cell phenotype and ligand expression in patient cohorts, functional NK cell assays against B7-H6+ target cells\",\n      \"journal\": \"AIDS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — patient cohort plus in vitro functional assays establishing the mechanism; single lab, multiple readouts\",\n      \"pmids\": [\"30325780\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"B7-H6 promotes NHL cell proliferation, migration, and invasion via the Ras/MEK/ERK signaling pathway. B7-H6 knockdown reduced phosphorylation of Ras/MEK/ERK components, and the MEK inhibitor AZD8330 partially reversed proliferation and completely reversed migration/invasion induced by B7-H6 overexpression.\",\n      \"method\": \"siRNA/shRNA knockdown, quantitative phosphoproteomics, Western blot, B7-H6 overexpression, MEK inhibitor treatment, in vivo xenograft model, CCK-8 and Transwell assays\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phosphoproteomics plus genetic and pharmacologic validation of the Ras/MEK/ERK pathway; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"32606790\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Soluble B7-H6 inhibits NK cell-mediated target cell killing and is associated with diminished cell surface expression of NKG2D and NCRs (NKp30, NKp40, NKp46). B7-H6 is only marginally released in association with extracellular vesicles (EVs), and EVs from B7-H6-expressing cells do not stimulate NK cell-mediated killing.\",\n      \"method\": \"Recombinant protein production in HEK293 cells, NK cell killing assays, flow cytometry, nano flow cytometry (NanoFCM) of EVs, ultracentrifugation fractionation\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (functional assays, EV fractionation, NanoFCM) in single lab study\",\n      \"pmids\": [\"33671836\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"BRD4 acts as an epigenetic activator of B7-H6 transcription in AML cells. Histone acetylation mediated by CBP30/P300 facilitates BRD4 binding to the B7-H6 promoter, which recruits P-TEFb elongation factor to phosphorylate RNA polymerase II and activate B7-H6 transcription. BRD4 also co-binds with JMJD6 at a distal enhancer of the B7-H6 gene. Metformin modifies acetylation at the B7-H6 promoter, impairing BRD4 binding.\",\n      \"method\": \"BRD4 inhibition, ChIP, reporter assays, siRNA knockdown, metformin treatment, Western blot, flow cytometry, apoptosis assays\",\n      \"journal\": \"Oncoimmunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — ChIP mapping of BRD4 binding at B7-H6 promoter and enhancer plus functional validation with inhibitors and siRNA; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"33796404\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"All-trans retinoic acid (atRA) induces leukemia cell resistance to NK cell cytotoxicity by downregulating B7-H6 expression via reduction of c-Myc, which in turn inhibits B7-H6 transcription. DNA methylation and HDAC inhibitors could not restore B7-H6 mRNA, indicating epigenetic modifications are not responsible; the effect was specifically dependent on c-Myc signaling.\",\n      \"method\": \"CRISPR/Cas9 B7-H6 knockout, siRNA c-Myc knockdown, real-time PCR, flow cytometry, Western blotting, NK cell cytotoxicity assays\",\n      \"journal\": \"Cancer communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockouts/knockdowns plus pharmacologic epistasis ruling out alternative mechanisms; single lab, multiple orthogonal approaches\",\n      \"pmids\": [\"34236140\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"B7-H6 contains 6 functional N-linked glycosylation sites. N43 and N208 glycosylation are conserved in jawed vertebrates and are essential for NK cell activation by B7-H6. Mechanistically, N208 glycosylation contributes to membrane stability and prevents B7-H6 shedding, while N43 glycosylation is required for B7-H6/NKp30 binding affinity.\",\n      \"method\": \"PNGase F digestion, tunicamycin blockade, single-site mutagenesis, SDS-PAGE, flow cytometry, NK cell activation assays, phylogenetic and structural analysis\",\n      \"journal\": \"Frontiers in immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — site-directed mutagenesis of each glycosylation site plus functional assays establishing distinct mechanistic roles for N43 vs N208; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"38035117\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"SNRPA promotes HCC cell proliferation and lenvatinib resistance via B7-H6 by facilitating B7-H6 pre-mRNA maturation: SNRPA binds B7-H6 pre-mRNA directly and contributes to its intron 2 splicing. B7-H6 in turn activates STAT3/AKT signaling, promoting G1-S cell cycle transition and inhibiting apoptosis.\",\n      \"method\": \"Nanopore full-length cDNA sequencing, RNA-binding protein immunoprecipitation (RIP-seq), exon-exon/exon-intron junction PCR, siRNA/overexpression gain/loss-of-function, Western blot, flow cytometry, in vivo xenograft model\",\n      \"journal\": \"Bioscience trends\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RIP-seq plus splice-junction PCR plus gain/loss-of-function experiments; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"40240166\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"G-CSF derived from gastric cancer tumor microenvironment induces B7-H6 expression on tumor-infiltrating neutrophils via NF-κB signaling pathway activation. Blockade of B7-H6 on neutrophils promotes their apoptosis and shortens their lifespan.\",\n      \"method\": \"Flow cytometry, in vitro neutrophil stimulation with recombinant G-CSF and tumor supernatants, NF-κB pathway inhibition, apoptosis assays\",\n      \"journal\": \"Neoplasia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro stimulation with mechanistic pathway identification (NF-κB) plus functional blockade assay; single lab, multiple readouts\",\n      \"pmids\": [\"40054066\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CRISPR knockout of B7-H6 in CD4+ T cells reduced NK cell responses to HIV-1-infected cells in some donors, establishing that B7-H6 on HIV-infected cells contributes to NKp30-mediated NK recognition of HIV-infected cells.\",\n      \"method\": \"CRISPR editing of CD4+ T cells to knockout B7-H6, NK cell activation/response assays, ligand expression profiling by flow cytometry\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — CRISPR genetic knockout with functional NK readout; single study, preprint, noted donor variability\",\n      \"pmids\": [\"bio_10.1101_2024.06.24.600449\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"NKp30-B7-H6 interaction in HBV-related acute-on-chronic liver failure (ACLF) triggers NK cell upregulation of IL-32, which in turn induces apoptosis of hepatocytes (Huh7 cells) in a dose-dependent manner. In vitro NK-92 cells killed high-B7-H6-expressing Huh7 and B7-H6-transfected LO2 hepatocytes in an NKp30/B7-H6-dependent manner.\",\n      \"method\": \"Co-culture assays, CCK8 cytotoxicity assay, anti-NKp30 antibody stimulation, Annexin V/PI apoptosis assay, flow cytometry, immunochemistry in patient samples\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic in vitro assays with antibody blockade and transfection establishing NKp30-B7-H6-IL-32 axis; single lab, multiple readouts\",\n      \"pmids\": [\"26241657\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"B7-H6 is preferentially expressed in glioma stem-like cells (GSLCs) compared to parental glioma cell lines. Knockdown of B7-H6 by siRNA in GSLCs inhibited cell proliferation with decreased c-Myc expression and inactivation of PI3K/Akt and ERK/MAPK signaling pathways. RNMT (RNA guanine-7 methyltransferase) was identified as a downstream target co-expressed with B7-H6 and c-Myc; RNMT expression was inhibited upon B7-H6 knockdown. CRISPR-Cas9 knockout of B7-H6 in 293T cells also suppressed proliferation.\",\n      \"method\": \"siRNA knockdown, CRISPR-Cas9 knockout, Western blot, TCGA co-expression analysis, flow cytometry, proliferation assays\",\n      \"journal\": \"Journal of immunology research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — two genetic perturbation methods (siRNA + CRISPR) with pathway readouts; single lab\",\n      \"pmids\": [\"32322592\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"B7-H6 knockdown in glioma cell lines (HepG2 and SMMC-7721) decreased expression of C-myc, C-fos, and Cyclin-D1, and was accompanied by reduced cell proliferation, migration, invasion, and G1 cell cycle arrest, indicating B7-H6 regulates these oncogenic factors intracellularly.\",\n      \"method\": \"RNA interference (siRNA), CCK-8 proliferation assay, migration/invasion assays, Western blot, flow cytometry cell cycle analysis\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — siRNA knockdown with downstream expression readouts; single lab, no pathway rescue experiments\",\n      \"pmids\": [\"30186042\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Lipopolysaccharide (LPS) induces B7-H6 expression in glioma cell lines. B7-H6 knockdown increased E-cadherin and Bax expression while decreasing vimentin, N-cadherin, MMP-2, MMP-9, and survivin, suggesting B7-H6 promotes EMT and invasion-related signaling.\",\n      \"method\": \"LPS stimulation, siRNA knockdown, Western blot, cell viability/migration/invasion assays\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single siRNA knockdown with downstream protein expression readouts; single lab, no mechanistic pathway rescue\",\n      \"pmids\": [\"29679856\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"B7-H6 promotes migration and invasion of medulloblastoma (Daoy) cells by activating c-MYC, which in turn enhances F-actin cytoskeleton rearrangement and upregulates MMP-2 and MMP-9 expression.\",\n      \"method\": \"siRNA knockdown, plasmid overexpression, Transwell migration/invasion assays, Western blot, immunofluorescence staining of F-actin\",\n      \"journal\": \"Medical oncology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, siRNA/overexpression with functional readouts but no pathway rescue or genetic epistasis experiments\",\n      \"pmids\": [\"36692844\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"B7-H6 (NCR3LG1) is a B7-family transmembrane ligand whose extracellular V-like domain binds the activating NK receptor NKp30 (crystal structure solved at 2.5 Å) to trigger NK cell cytotoxicity and cytokine secretion; it is absent from normal tissues but induced on tumor cells and inflammatory monocytes/neutrophils (via TLR/cytokine and NF-κB pathways), regulated transcriptionally by c-Myc/N-Myc and the epigenetic reader BRD4 (via CBP/P300 acetylation), cleaved from the cell surface by ADAM10/ADAM17 metalloproteases to generate soluble B7-H6 that downregulates NKp30 on NK cells, requires N-linked glycosylation at N43 (for NKp30 binding) and N208 (for membrane stability) for full function, and is exploited by HCMV (via US18/US20-mediated endosomal degradation) and HIV-2 to evade NK surveillance; intracellularly, B7-H6 activates Ras/MEK/ERK, PI3K/Akt, and STAT3/AKT signaling to promote tumor cell proliferation, migration, and survival.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"B7-H6 (NCR3LG1) is a B7-family cell-surface ligand that links cellular stress and malignant transformation to innate immune surveillance by directly engaging the activating NK receptor NKp30 to trigger NK cell cytotoxicity and cytokine secretion [#0]. The structural basis of this interaction is an antibody-like engagement in which NKp30 uses both faces of its Ig-like domain to contact CDR-like loops of the B7-H6 V-like domain, a geometry distinct from the inhibitory CTLA-4\\u2013B7 and PD-1\\u2013PD-L complexes [#1]. B7-H6 is largely absent from normal tissue and is induced on tumor cells and on proinflammatory CD14+CD16+ monocytes and neutrophils following TLR ligation or proinflammatory cytokines, including a G-CSF/NF-\\u03baB axis on tumor-infiltrating neutrophils [#0, #3, #18]. Its expression is transcriptionally driven by c-Myc and N-Myc binding the promoter and by an epigenetic program in which CBP/P300-dependent histone acetylation recruits BRD4 to promoter and enhancer elements [#8, #14], and it is further upregulated by chemotherapy, radiation, and heat shock to sensitize tumor cells to NK lysis [#7]. Surface B7-H6 is shed by the metalloproteases ADAM10 and ADAM17 to generate soluble ligand that, through chronic NKp30 engagement, downregulates NKp30 and impairs NK cell function\\u2014a route exploited as immune escape in cancer and HIV-2 infection [#2, #5, #11, #13]. N-linked glycosylation governs function site-specifically: N43 glycosylation is required for NKp30-binding affinity, while N208 glycosylation stabilizes the protein at the membrane and prevents shedding [#16]. Beyond its role as an immune ligand, B7-H6 acts intracellularly to promote tumor cell proliferation, migration, and invasion through Ras/MEK/ERK, PI3K/Akt, and STAT3/AKT signaling, with c-Myc as a recurrent downstream effector [#12, #17, #21]. Viral antagonism is also documented: HCMV genes US18 and US20 target B7-H6 for endosomal degradation to evade NKp30-mediated recognition [#10].\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Established the existence and function of B7-H6, answering whether a dedicated tumor-restricted ligand drives NKp30 activation.\",\n      \"evidence\": \"Protein identification with binding and NK activation assays on human tumor cells\",\n      \"pmids\": [\"19528259\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve the structural basis of recognition\", \"Regulation of its tumor-restricted expression unknown\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined the structural basis of NKp30\\u2013B7-H6 recognition, showing an antibody-like engagement distinct from inhibitory B7-family complexes.\",\n      \"evidence\": \"X-ray crystallography of the NKp30\\u2013B7-H6 complex\",\n      \"pmids\": [\"21422170\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address post-translational requirements for binding\", \"Affinity modulation in vivo not resolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identified how B7-H6 expression is induced, linking it both to inflammatory cell activation and to chromatin-level transcriptional control.\",\n      \"evidence\": \"TLR/cytokine stimulation of monocytes/neutrophils with vesicle fractionation; HDAC2/3 knockdown with promoter ChIP and reporter assays\",\n      \"pmids\": [\"23687088\", \"23801635\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct transcription factors not yet identified\", \"Relationship between vesicle-associated and shed soluble forms unclear\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Showed that surface B7-H6 is proteolytically shed, explaining the origin of soluble ligand and a mechanism limiting NK activation.\",\n      \"evidence\": \"ADAM10/ADAM17 pharmacologic inhibition and siRNA with flow cytometry, NK assays, and patient serum ELISA\",\n      \"pmids\": [\"24780758\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not establish functional consequences of soluble form on NK receptors\", \"Cleavage site not mapped\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Demonstrated that chronic NKp30 engagement by soluble or surface B7-H6 downregulates NKp30 and other NK receptors, defining a self-limiting immune-escape circuit.\",\n      \"evidence\": \"Patient-derived NK cell phenotyping and functional assays across ovarian carcinoma, neuroblastoma, and HBV-related liver failure cohorts\",\n      \"pmids\": [\"26137398\", \"25877893\", \"26241657\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism of NKp30 downregulation not defined\", \"Reversibility of receptor suppression untested\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Established B7-H6 as the mediator of therapy-induced NK sensitivity, connecting cellular stress to immune recognition.\",\n      \"evidence\": \"Chemotherapy/radiation/heat-shock treatment with B7-H6 shRNA knockdown and NK cytotoxicity assays\",\n      \"pmids\": [\"26472927\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Stress-responsive transcriptional sensors not identified\", \"Single-lab finding\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identified c-Myc/N-Myc as direct transcriptional drivers of B7-H6 and showed that anti-B7-H6 antibodies can block activation sterically rather than competitively.\",\n      \"evidence\": \"Promoter ChIP, reporter assays, Myc knockdown/inhibition; crystallography of 17B1.3\\u2013B7-H6 with mutagenesis\",\n      \"pmids\": [\"27622013\", \"27663271\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether B7-H6 dimerizes physiologically remained unresolved\", \"Myc cofactors at the promoter not mapped\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Revealed viral antagonism of B7-H6, showing HCMV degrades the ligand to evade NK surveillance.\",\n      \"evidence\": \"HCMV infection with US18/US20 gene deletion, flow cytometry, endosomal degradation analysis\",\n      \"pmids\": [\"28819195\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular details of US18/US20-mediated targeting not fully resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended the immune-escape paradigm to retroviral infection, showing HIV-2-infected T cells express B7-H6 to chronically suppress NKp30.\",\n      \"evidence\": \"Patient cohort flow cytometry and NK functional assays against B7-H6+ targets\",\n      \"pmids\": [\"30325780\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Why HIV-2 induces rather than degrades B7-H6 not explained\", \"Selective loss of cytolysis but not IFN-\\u03b3 unexplained\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined the epigenetic activation program and intracellular oncogenic signaling of B7-H6, beyond its ligand role.\",\n      \"evidence\": \"BRD4/CBP-P300 ChIP and inhibitor studies in AML; Ras/MEK/ERK phosphoproteomics and inhibitor rescue in lymphoma; atRA/c-Myc epistasis in leukemia\",\n      \"pmids\": [\"33796404\", \"32606790\", \"34236140\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a surface ligand drives intracellular kinase signaling mechanistically unclear\", \"Receptor/effector linking B7-H6 to Ras/MEK/ERK not identified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolved site-specific roles of N-glycosylation, separating ligand-binding competence (N43) from membrane stability and shedding resistance (N208).\",\n      \"evidence\": \"Single-site mutagenesis, PNGase F/tunicamycin treatment, NK activation assays, phylogenetic analysis\",\n      \"pmids\": [\"38035117\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Roles of the other four glycosylation sites undefined\", \"Glycan structures themselves not characterized\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected B7-H6 expression to RNA processing and downstream STAT3/AKT-driven proliferation, broadening its tumor-intrinsic functions.\",\n      \"evidence\": \"SNRPA RIP-seq and splice-junction PCR with gain/loss-of-function in HCC; G-CSF/NF-\\u03baB induction in neutrophils with apoptosis assays\",\n      \"pmids\": [\"40240166\", \"40054066\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct receptor mediating intracellular signaling unknown\", \"Generalizability across tumor types untested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How a cell-surface ligand transduces the intracellular Ras/MEK/ERK, PI3K/Akt, and STAT3/AKT signals attributed to B7-H6 remains the central unresolved mechanistic question.\",\n      \"evidence\": \"No discovery in the corpus identifies a cytoplasmic effector or receptor coupling B7-H6 to these pathways\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No defined intracellular signaling partner\", \"Distinction between cell-autonomous and NKp30-engagement effects unresolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 2, 16]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [2, 3, 13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0, 1, 5]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [12, 17, 21]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [8, 14]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"NKp30\", \"ADAM10\", \"ADAM17\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":8,"faith_total":8,"faith_pct":100.0}}