{"gene":"ADAM8","run_date":"2026-06-09T22:02:41","timeline":{"discoveries":[{"year":2002,"finding":"ADAM8 undergoes autocatalytic prodomain removal: mutation of Glu330 to Gln in the Zn2+-binding motif (HExxH) completely blocks propeptide cleavage. The ectodomain (but not the isolated MP domain) can trans-cleave the propeptide from catalytically inactive ADAM8. Active soluble ADAM8 cleaves myelin basic protein and a fluorogenic peptide substrate, inhibited by batimastat (IC50 ~50 nM) but not by TIMPs 1–4. A remnant form lacking the MP domain mediates cell adhesion via the disintegrin/Cys-rich domain, blocked by anti-disintegrin antibody.","method":"Site-directed mutagenesis (E330Q), co-transfection rescue assay, in vitro protease assay with purified enzyme, cell adhesion assay with substrate-bound recombinant protein","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstitution, active-site mutagenesis, and multiple orthogonal functional assays in one rigorous study","pmids":["12372841"],"is_preprint":false},{"year":2002,"finding":"Soluble recombinant ADAM8 is an active metalloprotease in vitro that hydrolyzes myelin basic protein and peptide substrates derived from known metalloprotease cleavage sites of membrane-bound cytokines/receptors. Unlike ADAM10, 12, 17 and MT-MMPs, ADAM8 activity is not inhibited by any of the four TIMPs (TIMP-1 to -4), but is inhibited by hydroxamate-based inhibitors.","method":"In vitro protease assay with purified recombinant ADAM8; TIMP inhibition assay","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct in vitro enzymatic reconstitution, replicated by multiple independent groups","pmids":["12135759"],"is_preprint":false},{"year":2003,"finding":"ADAM8 catalyzes ectodomain shedding of CD23 (the low-affinity IgE receptor FcεRII): proteolytically inactive ADAM8 (E330Q mutant) does not release soluble CD23, and a physical association between ADAM8 and membrane-bound CD23 was detected. ADAM8 also has distinct substrate specificity from ADAM17 on synthetic peptide libraries.","method":"Soluble CD23 release assay with active vs. inactive (E330Q) ADAM8; co-immunoprecipitation of ADAM8 with membrane CD23; peptide substrate library screening","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — active-site mutant controls plus physical interaction data, replicated concept across labs","pmids":["12777399"],"is_preprint":false},{"year":2004,"finding":"ADAM8 cleaves the neural cell adhesion molecule CHL1 at two sites in its extracellular fibronectin domains (releasing 125 kDa and 165 kDa fragments), inhibited by batimastat. The inactive E330Q-ADAM8 mutant does not cleave CHL1, while active ADAM10 and ADAM17 also fail to cleave CHL1, establishing ADAM8 substrate specificity. In ADAM8-deficient mice, CHL1 processing in brain extracts is almost undetectable. Processed CHL1 promotes neurite outgrowth and suppresses neuronal cell death in co-culture assays; these effects are absent with the inactive E330Q mutant.","method":"In vitro cleavage of CHL1-Fc fusion protein; transfection with WT vs. E330Q ADAM8; brain extracts from Adam8-/- mice; co-culture neurite outgrowth and apoptosis assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — in vitro reconstitution, active-site mutant, knockout mouse validation, and functional cellular readout","pmids":["14761956"],"is_preprint":false},{"year":2006,"finding":"Soluble ADAM8 (pro- + metalloprotease domain, autoactivated by autocatalytic prodomain removal) cleaves peptides representing extracellular domain sequences of 14 membrane proteins involved in inflammation and neurodegeneration (including amyloid precursor protein APP) out of 34 tested. Full-length APP is cleaved by active ADAM8 in HEK293 cells with similar efficiency to ADAM10; inactive ADAM8 does not cleave APP. Amino acid substitution analysis of the MBP cleavage sequence defined sequence criteria for ADAM8 cleavage.","method":"ProteaseSpot peptide cleavage assay with E. coli-expressed recombinant ADAM8; APP cleavage in HEK293 cells with WT vs. inactive ADAM8; mutagenesis of cleavage sequence","journal":"Biological chemistry","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — in vitro enzymatic assay plus cell-based confirmation with mutant controls, single lab","pmids":["16542157"],"is_preprint":false},{"year":2007,"finding":"ADAM8 is constitutively present on the cell surface and in intracellular granules of human neutrophils. Upon activation, ADAM8 is mobilized from granules to the plasma membrane and then shed through a metalloproteinase-dependent mechanism. Cell-surface and soluble ADAM8 increase ectodomain shedding of membrane-bound L-selectin in mammalian cells.","method":"Subcellular fractionation, immunofluorescence, confocal imaging of granule mobilization; L-selectin shedding assay in ADAM8-expressing cells","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — direct localization by fractionation/imaging tied to functional shedding assay, single lab","pmids":["17548643"],"is_preprint":false},{"year":2001,"finding":"The cysteine-rich/disintegrin domain of ADAM8 is responsible for stimulating osteoclast (OCL) formation and bone-resorbing activity. Conditioned media from cells expressing a secretable form of ADAM8 increased OCL formation dose-dependently; antisense oligonucleotides to ADAM8 inhibited OCL formation; truncation analysis mapped the OCL-stimulatory activity to the disintegrin/Cys-rich domain.","method":"Antisense oligonucleotide knockdown in mouse bone marrow cultures; conditioned media from ADAM8-transfected cells; truncation/domain mapping constructs; pit formation assay","journal":"Journal of bone and mineral research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain mapping by truncation constructs plus functional loss-of-function, single lab","pmids":["11341326"],"is_preprint":false},{"year":2009,"finding":"ADAM8 activation requires a novel pre-processing step: before terminal autocatalytic prodomain removal, the prodomain is first cleaved at Glu158 (N-terminal sequencing data), fracturing the pro-segment prior to removal of the cysteine switch (Cys167). This pre-processing requires the disintegrin (DIS) and cysteine-rich/EGF (CR/EGF) domains — ADAM8 constructs lacking these domains cannot complete pre-processing. Without pre-processing, the pro-segment re-associates with the catalytic domain and strongly inhibits activity.","method":"N-terminal sequencing of activation intermediates; enterokinase-cleavable construct engineering to bypass pre-processing; domain-deletion constructs; specific activity measurements","journal":"Bioscience reports","confidence":"High","confidence_rationale":"Tier 1 / Moderate — N-terminal sequencing of cleavage products plus engineered constructs and domain deletions in one study","pmids":["18811590"],"is_preprint":false},{"year":2014,"finding":"N-glycosylation at four sites (Asn-67, Asn-91, Asn-436, Asn-612) is required for correct ADAM8 processing, localization, stability, and activity. Asn-91 glycosylation (high mannose, prodomain) is essential for exit from the ER; Asn-612 (complex type, remnant form) for exit from the Golgi and cell-surface localization; Asn-436 mutation leads to enhanced lysosomal degradation. ADAM8 dimerization on the cell surface was detected specifically in ERα-negative (but not ERα-positive) breast cancer cells and required N-glycosylation.","method":"Site-directed mutagenesis of four Asn sites; glycosidase treatment; subcellular fractionation; Western blot for processing; flow cytometry for surface localization; glycan analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — mutagenesis of all four sites with orthogonal localization and processing assays in one study","pmids":["25336660"],"is_preprint":false},{"year":2015,"finding":"ADAM8 requires multimerization for biological function and associates with β1 integrin on the cell surface of pancreatic cancer (PDAC) cells. A peptidomimetic inhibitor (BK-1361), designed by structural modelling of the disintegrin domain, prevents ADAM8 multimerization and reduces PDAC cell invasiveness, ERK1/2 activation, and MMP activity. In vivo, BK-1361 decreased tumor burden and metastasis in mouse models of PDAC.","method":"Co-immunoprecipitation (ADAM8 with β1 integrin); structural modelling of disintegrin domain for inhibitor design; invasion assays; ERK1/2 and MMP activity assays; mouse xenograft and Kras(G12D) PDAC model","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP, cell-based functional assays, pharmacological inhibition, and in vivo validation across multiple models","pmids":["25629724"],"is_preprint":false},{"year":2013,"finding":"In triple-negative breast cancer (TNBC), ADAM8 promotes angiogenesis through release of VEGF-A and transendothelial cell migration via β1-integrin activation. ADAM8 knockdown tumors in an orthotopic mouse model failed to grow beyond palpable size and showed poor vascularization, reduced circulating tumor cells, and reduced brain metastases. Anti-ADAM8 antibody treatment reduced primary tumor burden and metastases in a resection model.","method":"shRNA knockdown; orthotopic mouse model; antibody treatment in vivo; VEGF-A release assay; β1-integrin activation assay; transendothelial migration assay","journal":"EMBO molecular medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function with defined molecular readouts, in vivo validation, and pharmacological rescue, replicated with orthogonal approaches","pmids":["24375628"],"is_preprint":false},{"year":2015,"finding":"ADAM8 mediates temozolomide (TMZ) chemoresistance in glioblastoma (GBM) cells by enhancing pAkt/PI3K and pERK1/2 signaling. Soluble HGF receptor/c-met and CD44 were identified as metalloprotease substrates in TMZ-treated GBM cells by proteomics; blocking HGF R/c-met prevented TMZ-induced invasiveness. ADAM8 knockdown or pharmacological inhibition (with BB-94 but not the ADAM8-sparing BB-2516) increased TMZ sensitivity.","method":"ADAM8 shRNA knockdown, overexpression, and pharmacological inhibition; proteomics substrate identification; blocking antibodies for c-met; pAkt/pERK immunoblotting; Matrigel invasion assay","journal":"Neuro-oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple approaches (KD, OE, pharmacological inhibition, proteomics substrates) in single lab","pmids":["25825051"],"is_preprint":false},{"year":2017,"finding":"ADAM8 cytoplasmic domain is required for MMP-9 upregulation: re-expression of WT ADAM8 (but not ADAM8 lacking the cytoplasmic domain) in ADAM8-knockdown breast cancer cells restored elevated pERK1/2, pCREB(S133), and MMP-9 transcription. ADAM8 also sheds PSGL-1 from breast cancer cells, and antibodies against PSGL-1 reduced transendothelial migration, linking ADAM8-mediated PSGL-1 shedding to transendothelial migration.","method":"Re-expression of WT vs. cytoplasmic-domain-deleted ADAM8; pERK/pCREB immunoblotting; qPCR for MMP-9; anti-PSGL-1 antibody blocking in transendothelial migration assay","journal":"International journal of cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain-deletion rescue experiment plus antibody blocking, single lab","pmids":["28986926"],"is_preprint":false},{"year":2010,"finding":"ADAM8 deficiency in mice completely prevents development of ovalbumin-induced airway inflammation and hyperresponsiveness. ADAM8 expression is required in both hematopoietic cells (including T cells) and non-hematopoietic cells for full asthma manifestation, demonstrated by bone marrow chimera experiments. Loss of ADAM8 impaired migration of T cells, eosinophils, and myeloid cells from blood vessels to the lung.","method":"Adam8-/- knockout mice; OVA-induced asthma model; bone marrow chimera experiments; bronchoalveolar lavage analysis; histology; airway hyperresponsiveness measurement","journal":"American journal of respiratory and critical care medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — knockout and chimera experiments with defined cellular and functional phenotype, replicated across multiple model conditions","pmids":["20194813"],"is_preprint":false},{"year":2013,"finding":"ADAM8 limits allergic airway inflammation (AAI) by activating the intrinsic apoptosis pathway in myeloid leukocytes (eosinophils and macrophages): Adam8-/- mice have fewer apoptotic eosinophils and macrophages in airways during AAI, and Adam8-/- macrophages and eosinophils show reduced apoptosis when the intrinsic (but not the extrinsic) apoptosis pathway is triggered in vitro. Leukocyte-derived ADAM8 predominantly mediates this anti-inflammatory activity, confirmed by bone marrow chimeras.","method":"Adam8-/- knockout mice; bone marrow chimera; OVA and house dust mite AAI models; apoptosis assays (intrinsic vs. extrinsic pathway activation) in isolated cells; airway leukocyte counting","journal":"Journal of immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO mice, chimera experiments, and cell-specific in vitro apoptosis assays with pathway discrimination","pmids":["23670189"],"is_preprint":false},{"year":2005,"finding":"ADAM8-deficient (Adam8-/-) mice develop normally without major defects in any tissue or organ, indicating ADAM8 is dispensable for normal development and homeostasis. Expression analysis showed ADAM8 is prominently expressed during mouse development in decidua, trophoblast derivatives, gonadal ridge, thymus, cartilage/bone, brain, spinal cord, and perivenous mesenchyme.","method":"Targeted gene deletion (Adam8-/- mice); developmental expression analysis by in situ hybridization/immunostaining; histological phenotyping","journal":"Developmental dynamics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — comprehensive knockout phenotyping and expression mapping, single lab","pmids":["15580619"],"is_preprint":false},{"year":2005,"finding":"ADAM8-mediated shedding of VCAM-1 from endothelial cells reduces eosinophil adhesion (via α4β1 integrin) and suppresses experimental asthma: ADAM8-transgenic mice (expressing ectodomain of CD156/ADAM8) show markedly reduced cellular infiltrates in peribronchovascular lesions compared to controls, associated with increased VCAM-1 shedding.","method":"ADAM8 transgenic mice; OVA-induced asthma model; VCAM-1 shedding assay in human umbilical vein endothelial cells stimulated with ADAM8-transgenic cells","journal":"Immunology letters","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — transgenic mouse model with defined substrate (VCAM-1 shedding) and in vitro endothelial cell assay, single lab","pmids":["16154205"],"is_preprint":false},{"year":2011,"finding":"ADAM8 overexpression in osteoclast (OCL) precursors (TRAP-ADAM8 transgenic mice) produces osteopenia, hypermultinucleated OCLs with increased bone resorption capacity, enhanced OCL precursor fusion, increased TRAF6 expression, and elevated NF-κB, Erk, and Akt signaling, as well as increased p-Pyk2 and p-Src activation. ADAM8 knockout mice do not display a basal bone phenotype but fail to increase RANKL production, OCL formation, or calvarial fibrosis in response to TNF-α.","method":"TRAP-ADAM8 transgenic overexpression mice; Adam8-/- knockout mice; TNF-α calvarial injection model; osteoclast formation and bone resorption assays; Western blot for TRAF6, NF-κB, Erk, Akt, p-Pyk2, p-Src","journal":"Journal of bone and mineral research","confidence":"High","confidence_rationale":"Tier 2 / Strong — complementary gain- and loss-of-function in vivo models with defined signaling pathway readouts","pmids":["20683884"],"is_preprint":false},{"year":2009,"finding":"Catalytically inactive ADAM8 (E330Q knock-in transgenic DBA/1J mice) shows decreased incidence and severity of collagen-induced arthritis with reduced synovial inflammation, cartilage degradation, and bone resorption, demonstrating that ADAM8 proteolytic activity is required for experimental arthritis development.","method":"Transgenic mice expressing E330Q catalytically inactive ADAM8; LPS-synchronized collagen-induced arthritis model; histological scoring of joints; expression profiling","journal":"Clinical and experimental immunology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — catalytically inactive knock-in with in vivo disease phenotype, single lab but rigorous genetic approach","pmids":["19737139"],"is_preprint":false},{"year":2010,"finding":"ADAM8 overexpression in endothelial cells increases ectodomain shedding of multiple pro-angiogenic membrane proteins including CD31, Tie-2, Flk-1, Flt-1, EphrinB2, EphB4, VE-cadherin, KL-1, E-selectin, and neuregulin-1β2. In Adam8-/- mice, the oxygen-induced retinopathy model shows more retinal re-vascularization but fewer neovascular tufts, and heterotopically injected tumor cells grow faster, suggesting ADAM8 limits pathological angiogenesis.","method":"ADAM8 overexpression in endothelial cells with ectodomain shedding assays; Adam8-/- mice in OIR model; tumor cell heterotopic injection model","journal":"Journal of molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell overexpression shedding assay plus knockout in vivo models, single lab","pmids":["20119708"],"is_preprint":false},{"year":2017,"finding":"ADAM8 promotes migration of IL-5-stimulated eosinophils on periostin-rich extracellular matrix: ADAM8 on eosinophils degrades (sheds the Stiny-1 epitope of) periostin in the extracellular matrix and is required for migration. Shed ADAM8 proteoforms lacking the cytoplasmic tail were detected in supernatants. ADAM8 antibodies and metalloproteinase inhibitors both blocked eosinophil migration and periostin epitope clearance.","method":"Fluorescence microscopy of eosinophil migration on periostin; anti-ADAM8 and metalloprotease inhibitor blocking; Western blot of ADAM8 proteoforms in supernatants","journal":"Clinical and experimental allergy","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — antibody blocking with defined substrate and functional readout, single lab","pmids":["28378503"],"is_preprint":false},{"year":2017,"finding":"Leukocytic ADAM8 is required for chemokine-induced chemotaxis and transendothelial and transepithelial migration of monocytic THP-1 cells, as well as αL-integrin upregulation and THP-1 adhesion to endothelial cells. On endothelial cells, ADAM8 enhances transendothelial migration and cytokine-induced permeability. In Adam8-/- mice, intranasal LPS-induced acute lung inflammation shows reduced leukocyte infiltration. Bone marrow macrophage transfer experiments confirmed a predominantly leukocyte-intrinsic promigratory function.","method":"shRNA knockdown in THP-1 cells; Adam8-/- knockout mice; LPS-induced acute lung inflammation model; BMDM transfer experiments; transendothelial/transepithelial migration assays; αL-integrin flow cytometry","journal":"American journal of physiology. Lung cellular and molecular physiology","confidence":"High","confidence_rationale":"Tier 2 / Strong — knockdown, knockout mouse, and adoptive transfer with defined functional phenotype across orthogonal assay systems","pmids":["28596294"],"is_preprint":false},{"year":2020,"finding":"ADAM8 regulates angiogenesis in glioblastoma via the JAK/STAT3 pathway controlling osteopontin (OPN) expression: ADAM8 knockdown in U87 glioma cells decreased angiogenesis and tumor volumes in vivo. In vitro, ADAM8 regulates OPN through JAK/STAT3 signaling in both glioma cells and primary macrophages.","method":"ADAM8 shRNA knockdown; stereotactic mouse brain injection model; in vitro angiogenesis assays; JAK/STAT3 pathway inhibition; OPN measurement","journal":"Biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockdown with in vivo validation and defined signaling pathway, single lab","pmids":["33544472"],"is_preprint":false},{"year":2022,"finding":"ADAM8 is present as an active protease in extracellular vesicles (EVs) from PDAC cells and is associated with lipocalin 2 (LCN2) and MMP-9 in an ADAM8-dependent manner (ADAM8 KO cells show reduced LCN2 and MMP-9 in EVs). ADAM8 sorting into EVs is independent of the TSG101 ESCRT-I pathway despite the presence of a PTAP recognition motif in ADAM8's cytoplasmic domain.","method":"ADAM8 CRISPR KO in PDAC cells; EV isolation and proteomics; Western blot for LCN2, MMP-9, TSG101 in EVs; co-culture with macrophages","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO with EV proteomics and functional co-culture, single lab","pmids":["35216088"],"is_preprint":false},{"year":2024,"finding":"In macrophages after myocardial infarction, ADAM8 binds to ANXA2 and promotes phosphorylation of ANXA2 at Ser26. ADAM8 knockout impedes ANXA2 phosphorylation, inhibits mTOR Ser2448 phosphorylation, and activates autophagy, leading to enhanced angiogenesis and suppressed inflammation. Macrophage-specific ADAM8 KO mice show improved cardiac repair, confirmed by bone marrow transplantation; these effects are reversed by ADAM8 overexpression.","method":"Macrophage-specific ADAM8 KO (CRISPR/Cas9, Lyz2-Cre); bone marrow transplantation; Co-IP/mass spectrometry (ADAM8-ANXA2 interaction); RNA sequencing; phospho-specific Western blots (p-mTOR, p-ANXA2); ANXA2 phosphorylation activation/inactivation experiments","journal":"Journal of advanced research","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — Co-IP/MS interaction data plus functional rescue experiments and genetic confirmation, single lab but multiple orthogonal methods","pmids":["39097092"],"is_preprint":false},{"year":1997,"finding":"Human CD156 (ADAM8) cDNA encodes a transmembrane glycoprotein with an extracellular region containing metalloprotease, disintegrin, cysteine-rich, and EGF-like domains homologous to hemorrhagic snake venom proteins. mRNA is expressed in macrophage cell lines, granulocytes, monocytes, and B cells but not T cell lines. The gene maps to human chromosome 10q26.3.","method":"cDNA cloning and sequencing from THP-1 macrophage and granulocyte libraries; Northern blot expression analysis; chromosomal mapping","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — cDNA sequencing and expression analysis establishing domain architecture and cell-type specificity, replicated across labs","pmids":["9126482"],"is_preprint":false},{"year":1997,"finding":"The murine CD156/ADAM8 gene spans ~14 kb, is composed of 24 exons, and encodes a protein with metalloprotease domain (exons 7–12), disintegrin domain (exons 12–15), and transmembrane region (single exon 19). The promoter contains cis-acting elements at positions −183, −334, and −623 and LPS-inducible elements at −183 and −390, and interferon-γ-inducible regions at −202, −507, and −659, defined by CAT reporter assays in monocytic cells.","method":"Genomic cloning, sequencing and exon mapping; chloramphenicol acetyltransferase (CAT) reporter assays with promoter deletion constructs in monocytic cell line; LPS and IFN-γ stimulation","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — promoter deletion analysis with reporter assays and stimulation experiments, single lab","pmids":["9218457"],"is_preprint":false},{"year":2019,"finding":"ADAM8 inhibition by hydroxamate-based inhibitors: BB-94 (batimastat), GW280264, FC387, and FC143 inhibit ADAM8 in vitro and in cell-based CD23 shedding assays; GM6001, TAPI2, and BB-2516 (marimastat) weakly inhibit ADAM8; GI254023 (ADAM10-specific) does not inhibit ADAM8. Structural modelling of the S1 position reveals T300 in ADAM8 (and T347 in ADAM17) vs. V327 in ADAM10 explains differential inhibitor selectivity.","method":"In vitro recombinant ADAM8 protease assay; cell-based CD23 shedding assay in HEK293 cells; structural modelling of inhibitor binding","journal":"Biological chemistry","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — in vitro enzymatic assay plus cell-based validation and structural modelling, single lab","pmids":["30738011"],"is_preprint":false},{"year":2022,"finding":"In glioblastoma, ADAM8 activates HB-EGF/EGFR signaling to upregulate CCL2 expression in tumor cells, which recruits tumor-associated macrophages (TAMs). TAMs in turn induce ADAM8 expression in GBM cells, forming a positive feedback loop that mediates TMZ chemoresistance. Only ADAM8 (and not other ADAM or MMP family members screened) was upregulated in GBM cells by macrophages under TMZ treatment.","method":"qPCR screen of ADAM/MMP family in co-culture; RNA-seq; ELISA and Western blot for CCL2; EGFR signaling analysis; in vitro and in vivo TAM recruitment assays; TCGA data analysis with IHC validation","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic co-culture screen plus pathway analysis and in vivo validation, single lab","pmids":["36230833"],"is_preprint":false}],"current_model":"ADAM8 is a membrane-anchored metalloprotease-disintegrin that undergoes obligatory autocatalytic prodomain removal (requiring a DIS/CR/EGF domain-coordinated pre-processing step at Glu158 before final cysteine-switch removal at Cys167, with N-glycosylation at Asn-91 and Asn-612 essential for correct ER/Golgi exit and cell-surface localization), after which it acts as a sheddase for multiple substrates including CD23, L-selectin, VCAM-1, CHL1, APP, PSGL-1, and angiogenic receptors; its disintegrin domain mediates cell adhesion and, through multimerization, associates with β1 integrin to activate ERK1/2, Akt/PI3K, and FAK signaling, while its cytoplasmic domain drives MMP-9 transcription via pERK/pCREB and in macrophages binds ANXA2 to regulate mTOR-autophagy, collectively enabling ADAM8 to control leukocyte migration, osteoclast differentiation and bone resorption, airway inflammation (with both pro- and anti-inflammatory roles depending on cell type), and cancer cell invasiveness, angiogenesis, and chemoresistance."},"narrative":{"mechanistic_narrative":"ADAM8 (CD156) is a membrane-anchored metalloprotease-disintegrin that couples regulated ectodomain shedding to adhesion-dependent signaling, controlling leukocyte migration, osteoclast-mediated bone resorption, angiogenesis, and tumor invasion [PMID:20194813, PMID:20683884, PMID:24375628, PMID:9126482]. Its protease domain is activated by an obligatory autocatalytic mechanism: a pre-processing cleavage at Glu158 that requires the disintegrin and cysteine-rich/EGF domains fractures the prodomain before final cysteine-switch removal at Cys167, and active-site mutation (E330Q) of the HExxH motif abolishes activity [PMID:12372841, PMID:18811590]. Correct maturation also depends on site-specific N-glycosylation, with Asn-91 required for ER exit and Asn-612 for Golgi exit and cell-surface display [PMID:25336660]. Unusually among ADAMs, ADAM8 is resistant to all four TIMPs but inhibited by hydroxamate-based compounds, a selectivity traced to an S1-pocket threonine (T300) [PMID:12135759, PMID:30738011]. Once active, ADAM8 sheds a broad substrate set including CD23, L-selectin, CHL1, APP, VCAM-1, PSGL-1, periostin, and pro-angiogenic receptors, generating products with distinct downstream consequences [PMID:12777399, PMID:14761956, PMID:17548643, PMID:16154205, PMID:28378503, PMID:20119708]. Beyond catalysis, the disintegrin domain mediates cell adhesion and, through multimerization, associates with β1 integrin to drive ERK1/2, Akt/PI3K, and downstream invasion programs, while the cytoplasmic domain is required for pERK/pCREB-dependent MMP-9 transcription [PMID:12372841, PMID:25629724, PMID:24375628, PMID:28986926]. In disease contexts ADAM8 promotes pancreatic, breast, and glioblastoma tumor invasion, angiogenesis, and chemoresistance [PMID:25629724, PMID:24375628, PMID:25825051], and is genetically required for allergic airway inflammation, arthritis, and inflammatory osteoclastogenesis, though it can also exert anti-inflammatory effects by promoting intrinsic apoptosis of myeloid leukocytes and, in macrophages, by binding ANXA2 to regulate mTOR-autophagy [PMID:20194813, PMID:19737139, PMID:20683884, PMID:23670189, PMID:39097092].","teleology":[{"year":1997,"claim":"Establishing ADAM8's domain architecture and expression pattern defined it as a candidate metalloprotease-disintegrin of myeloid lineage, framing all later mechanistic work.","evidence":"cDNA cloning/sequencing, Northern blot, and chromosomal mapping in human; genomic exon mapping and promoter reporter assays in mouse","pmids":["9126482","9218457"],"confidence":"Medium","gaps":["Did not demonstrate catalytic activity or identify substrates","Promoter elements mapped only in monocytic cells"]},{"year":2001,"claim":"Mapping osteoclast-stimulatory activity to the disintegrin/cysteine-rich domain showed ADAM8 has non-proteolytic adhesion/signaling functions in bone biology.","evidence":"Antisense knockdown, secretable-form conditioned media, and truncation constructs in mouse bone marrow cultures with pit formation assay","pmids":["11341326"],"confidence":"Medium","gaps":["Receptor/ligand partner for the disintegrin domain not identified","Single-lab loss-of-function"]},{"year":2002,"claim":"Reconstitution with active-site mutants established ADAM8 as a genuine autocatalytically activated metalloprotease distinct from TIMP-regulated ADAMs/MMPs.","evidence":"E330Q active-site mutagenesis, co-transfection trans-cleavage rescue, in vitro protease assays with purified enzyme, and TIMP/hydroxamate inhibition tests","pmids":["12372841","12135759"],"confidence":"High","gaps":["Physiological substrates not yet defined","Structural basis of TIMP resistance unresolved at this stage"]},{"year":2003,"claim":"Identification of CD23 as a shed substrate connected ADAM8 catalysis to a defined immune-relevant cleavage event.","evidence":"Soluble CD23 release with active vs. E330Q ADAM8, co-IP of ADAM8 with membrane CD23, and peptide library specificity profiling","pmids":["12777399"],"confidence":"High","gaps":["In vivo CD23 shedding by ADAM8 not demonstrated","Recognition determinants only partially mapped"]},{"year":2004,"claim":"CHL1 was established as a specific ADAM8 substrate with a functional neural readout, distinguishing ADAM8 from ADAM10/17.","evidence":"In vitro CHL1-Fc cleavage, WT vs. E330Q transfection, Adam8-/- brain extracts, and co-culture neurite/apoptosis assays","pmids":["14761956"],"confidence":"High","gaps":["Physiological neural phenotype of impaired CHL1 processing not characterized in vivo","Receptor mediating processed-CHL1 effects unknown"]},{"year":2006,"claim":"Profiling substrate peptides and APP cleavage broadened the substrate repertoire and defined sequence criteria for ADAM8 cleavage.","evidence":"ProteaseSpot peptide cleavage with recombinant ADAM8, APP cleavage in HEK293 with WT vs. inactive enzyme, and cleavage-site mutagenesis","pmids":["16542157"],"confidence":"Medium","gaps":["Many peptide hits not validated as full-length substrates in vivo","Single-lab study"]},{"year":2007,"claim":"Neutrophil studies showed ADAM8 is granule-stored, activation-mobilized to the surface, and itself shed, linking its trafficking to leukocyte function.","evidence":"Subcellular fractionation, confocal imaging of granule mobilization, and L-selectin shedding assay","pmids":["17548643"],"confidence":"Medium","gaps":["Mechanism triggering granule mobilization not defined","L-selectin shedding shown in heterologous cells"]},{"year":2009,"claim":"Defining a DIS/CR-EGF-dependent pre-processing step at Glu158 prior to Cys167 removal explained how ADAM8 escapes prodomain auto-inhibition.","evidence":"N-terminal sequencing of intermediates, enterokinase-bypass constructs, and domain-deletion specific-activity measurements","pmids":["18811590"],"confidence":"High","gaps":["Whether pre-processing is intra- or intermolecular not fully resolved","Trafficking compartment of each step not localized"]},{"year":2005,"claim":"Knockout and transgenic mouse models established ADAM8 as dispensable for development but functionally required and substrate-directed (VCAM-1) in inflammatory contexts.","evidence":"Adam8-/- phenotyping with developmental expression mapping; ADAM8-transgenic mice in OVA asthma with VCAM-1 shedding assays","pmids":["15580619","16154205"],"confidence":"Medium","gaps":["Functional redundancy masking developmental roles not excluded","VCAM-1 shedding study single-lab and transgenic-driven"]},{"year":2010,"claim":"Genetic and chimera experiments placed ADAM8 as required for allergic airway inflammation through promigratory roles in both hematopoietic and non-hematopoietic compartments, while also limiting pathological angiogenesis.","evidence":"Adam8-/- mice in OVA asthma with bone marrow chimeras; ADAM8 overexpression shedding assays plus Adam8-/- OIR and tumor models","pmids":["20194813","20119708"],"confidence":"High","gaps":["Causal substrate(s) mediating the migration defect not pinpointed","Pro- vs anti-angiogenic balance context-dependence unresolved"]},{"year":2011,"claim":"Complementary gain- and loss-of-function osteoclast models linked ADAM8 to TRAF6/NF-κB/Erk/Akt signaling and inflammatory bone resorption.","evidence":"TRAP-ADAM8 transgenic and Adam8-/- mice with TNF-α calvarial model and pathway immunoblots","pmids":["20683884"],"confidence":"High","gaps":["Direct molecular link from ADAM8 to TRAF6 induction not defined","Catalytic vs. adhesion contribution to osteoclast phenotype not dissected"]},{"year":2009,"claim":"A catalytically inactive E330Q knock-in established that ADAM8 proteolytic activity is required for experimental arthritis.","evidence":"E330Q knock-in DBA/1J mice in collagen-induced arthritis with histological scoring","pmids":["19737139"],"confidence":"High","gaps":["Relevant arthritic substrates not identified","Single-lab disease model"]},{"year":2013,"claim":"Studies in TNBC and AAI revealed a dual role: ADAM8 drives β1-integrin-dependent angiogenesis and metastasis in cancer yet limits airway inflammation by promoting intrinsic apoptosis of myeloid leukocytes.","evidence":"shRNA knockdown, orthotopic and resection mouse models, anti-ADAM8 antibody therapy, VEGF-A/transendothelial assays; Adam8-/- mice with chimeras and intrinsic vs extrinsic apoptosis assays","pmids":["24375628","23670189"],"confidence":"High","gaps":["Molecular basis of cell-type-dependent pro- vs anti-inflammatory switch unresolved","Apoptosis-promoting mechanism downstream of ADAM8 not defined"]},{"year":2014,"claim":"Site-specific N-glycosylation was shown to control ADAM8 ER/Golgi trafficking, stability, and surface dimerization, with dimerization selective to ERα-negative breast cancer cells.","evidence":"Mutagenesis of four Asn sites, glycosidase treatment, fractionation, flow cytometry, and glycan analysis","pmids":["25336660"],"confidence":"High","gaps":["Why dimerization is restricted to ERα-negative cells not mechanistically explained","Glycan-dependent partner interactions not mapped"]},{"year":2015,"claim":"Multimerization-dependent association with β1 integrin was shown to drive cancer invasion and was therapeutically targetable, while ADAM8 was implicated in glioblastoma chemoresistance via pAkt/pERK signaling.","evidence":"Co-IP with β1 integrin, structure-guided peptidomimetic BK-1361, invasion/ERK/MMP assays and PDAC mouse models; GBM knockdown/overexpression/inhibition with proteomic substrate ID (c-met, CD44)","pmids":["25629724","25825051"],"confidence":"High","gaps":["Whether β1-integrin signaling requires catalytic activity not fully separated","GBM substrate-to-resistance causal chain partly correlative"]},{"year":2017,"claim":"Cytoplasmic-domain and leukocyte-migration studies dissected adhesion/signaling functions: the cytoplasmic tail drives pERK/pCREB-dependent MMP-9 transcription and PSGL-1 shedding, and leukocytic ADAM8 is intrinsically required for chemotaxis and transmigration.","evidence":"WT vs. cytoplasmic-deletion rescue, qPCR/immunoblot, anti-PSGL-1 blocking; THP-1 knockdown, Adam8-/- LPS lung model, BMDM transfer, and αL-integrin/migration assays; eosinophil migration on periostin with ADAM8/protease blocking","pmids":["28986926","28596294","28378503"],"confidence":"High","gaps":["Direct effectors binding the cytoplasmic tail to drive ERK/CREB not all identified","Relative weight of shedding vs. integrin signaling in migration unresolved"]},{"year":2022,"claim":"Tumor microenvironment and vesicle studies extended ADAM8's reach: it forms an HB-EGF/EGFR–CCL2–TAM feedback loop in glioblastoma and is packaged with LCN2/MMP-9 into extracellular vesicles via a TSG101-independent route.","evidence":"Co-culture qPCR screen, RNA-seq, CCL2/EGFR analyses, TAM recruitment models; CRISPR KO PDAC cells with EV proteomics and macrophage co-culture","pmids":["36230833","35216088"],"confidence":"Medium","gaps":["EV sorting mechanism replacing ESCRT-I not identified","In vivo relevance of EV-borne ADAM8 not established"]},{"year":2024,"claim":"A non-proteolytic intracellular function emerged: macrophage ADAM8 binds and phosphorylates ANXA2 to sustain mTOR signaling and suppress autophagy, impairing post-infarct cardiac repair.","evidence":"Macrophage-specific ADAM8 KO, bone marrow transplantation, Co-IP/MS, RNA-seq, and phospho-specific immunoblots (p-ANXA2, p-mTOR)","pmids":["39097092"],"confidence":"High","gaps":["How a membrane sheddase accesses cytoplasmic ANXA2 to drive its phosphorylation not mechanistically explained","Single-lab study"]},{"year":null,"claim":"How ADAM8 selects between its proteolytic (sheddase), adhesive (disintegrin/β1-integrin), and intracellular (cytoplasmic-tail/ANXA2) modes in a cell-type-specific manner — and how this dictates its opposing pro- versus anti-inflammatory and pro- versus anti-angiogenic outcomes — remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No high-resolution structure of full-length or multimerized ADAM8","Unifying model integrating catalysis, integrin signaling, and intracellular partners absent","Determinants of context-dependent functional polarity unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1,2,3,4,5]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,1,27]},{"term_id":"GO:0098631","term_label":"cell adhesion mediator activity","supporting_discovery_ids":[0,6,9]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[9,10,24]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[5,8,9]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[5,23]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[8]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[8]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[13,14,21]},{"term_id":"R-HSA-1474244","term_label":"Extracellular matrix organization","supporting_discovery_ids":[3,20]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[9,17,24]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[9,10,11,28]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,7,8]}],"complexes":[],"partners":["ITGB1","ANXA2","CD23","CHL1","PSGL1","LCN2","MMP9"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P78325","full_name":"Disintegrin and metalloproteinase domain-containing protein 8","aliases":["Cell surface antigen MS2"],"length_aa":824,"mass_kda":88.8,"function":"Possible involvement in extravasation of leukocytes","subcellular_location":"Membrane","url":"https://www.uniprot.org/uniprotkb/P78325/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ADAM8","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ADAM8","total_profiled":1310},"omim":[{"mim_id":"610200","title":"MITOCHONDRIAL RIBOSOMAL PROTEIN L13; 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human ADAM8: a novel pre-processing step is required for catalytic activity.","date":"2009","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/18811590","citation_count":19,"is_preprint":false},{"pmid":"28691656","id":"PMC_28691656","title":"Mutational analysis of the MS2 lysis protein L.","date":"2017","source":"Microbiology (Reading, England)","url":"https://pubmed.ncbi.nlm.nih.gov/28691656","citation_count":18,"is_preprint":false},{"pmid":"35141043","id":"PMC_35141043","title":"Improved alpharetrovirus-based Gag.MS2 particles for efficient and transient delivery of CRISPR-Cas9 into target cells.","date":"2022","source":"Molecular therapy. 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of Glu330 to Gln in the Zn2+-binding motif (HExxH) completely blocks propeptide cleavage. The ectodomain (but not the isolated MP domain) can trans-cleave the propeptide from catalytically inactive ADAM8. Active soluble ADAM8 cleaves myelin basic protein and a fluorogenic peptide substrate, inhibited by batimastat (IC50 ~50 nM) but not by TIMPs 1–4. A remnant form lacking the MP domain mediates cell adhesion via the disintegrin/Cys-rich domain, blocked by anti-disintegrin antibody.\",\n      \"method\": \"Site-directed mutagenesis (E330Q), co-transfection rescue assay, in vitro protease assay with purified enzyme, cell adhesion assay with substrate-bound recombinant protein\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstitution, active-site mutagenesis, and multiple orthogonal functional assays in one rigorous study\",\n      \"pmids\": [\"12372841\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Soluble recombinant ADAM8 is an active metalloprotease in vitro that hydrolyzes myelin basic protein and peptide substrates derived from known metalloprotease cleavage sites of membrane-bound cytokines/receptors. Unlike ADAM10, 12, 17 and MT-MMPs, ADAM8 activity is not inhibited by any of the four TIMPs (TIMP-1 to -4), but is inhibited by hydroxamate-based inhibitors.\",\n      \"method\": \"In vitro protease assay with purified recombinant ADAM8; TIMP inhibition assay\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct in vitro enzymatic reconstitution, replicated by multiple independent groups\",\n      \"pmids\": [\"12135759\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"ADAM8 catalyzes ectodomain shedding of CD23 (the low-affinity IgE receptor FcεRII): proteolytically inactive ADAM8 (E330Q mutant) does not release soluble CD23, and a physical association between ADAM8 and membrane-bound CD23 was detected. ADAM8 also has distinct substrate specificity from ADAM17 on synthetic peptide libraries.\",\n      \"method\": \"Soluble CD23 release assay with active vs. inactive (E330Q) ADAM8; co-immunoprecipitation of ADAM8 with membrane CD23; peptide substrate library screening\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — active-site mutant controls plus physical interaction data, replicated concept across labs\",\n      \"pmids\": [\"12777399\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"ADAM8 cleaves the neural cell adhesion molecule CHL1 at two sites in its extracellular fibronectin domains (releasing 125 kDa and 165 kDa fragments), inhibited by batimastat. The inactive E330Q-ADAM8 mutant does not cleave CHL1, while active ADAM10 and ADAM17 also fail to cleave CHL1, establishing ADAM8 substrate specificity. In ADAM8-deficient mice, CHL1 processing in brain extracts is almost undetectable. Processed CHL1 promotes neurite outgrowth and suppresses neuronal cell death in co-culture assays; these effects are absent with the inactive E330Q mutant.\",\n      \"method\": \"In vitro cleavage of CHL1-Fc fusion protein; transfection with WT vs. E330Q ADAM8; brain extracts from Adam8-/- mice; co-culture neurite outgrowth and apoptosis assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — in vitro reconstitution, active-site mutant, knockout mouse validation, and functional cellular readout\",\n      \"pmids\": [\"14761956\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Soluble ADAM8 (pro- + metalloprotease domain, autoactivated by autocatalytic prodomain removal) cleaves peptides representing extracellular domain sequences of 14 membrane proteins involved in inflammation and neurodegeneration (including amyloid precursor protein APP) out of 34 tested. Full-length APP is cleaved by active ADAM8 in HEK293 cells with similar efficiency to ADAM10; inactive ADAM8 does not cleave APP. Amino acid substitution analysis of the MBP cleavage sequence defined sequence criteria for ADAM8 cleavage.\",\n      \"method\": \"ProteaseSpot peptide cleavage assay with E. coli-expressed recombinant ADAM8; APP cleavage in HEK293 cells with WT vs. inactive ADAM8; mutagenesis of cleavage sequence\",\n      \"journal\": \"Biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro enzymatic assay plus cell-based confirmation with mutant controls, single lab\",\n      \"pmids\": [\"16542157\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"ADAM8 is constitutively present on the cell surface and in intracellular granules of human neutrophils. Upon activation, ADAM8 is mobilized from granules to the plasma membrane and then shed through a metalloproteinase-dependent mechanism. Cell-surface and soluble ADAM8 increase ectodomain shedding of membrane-bound L-selectin in mammalian cells.\",\n      \"method\": \"Subcellular fractionation, immunofluorescence, confocal imaging of granule mobilization; L-selectin shedding assay in ADAM8-expressing cells\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — direct localization by fractionation/imaging tied to functional shedding assay, single lab\",\n      \"pmids\": [\"17548643\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"The cysteine-rich/disintegrin domain of ADAM8 is responsible for stimulating osteoclast (OCL) formation and bone-resorbing activity. Conditioned media from cells expressing a secretable form of ADAM8 increased OCL formation dose-dependently; antisense oligonucleotides to ADAM8 inhibited OCL formation; truncation analysis mapped the OCL-stimulatory activity to the disintegrin/Cys-rich domain.\",\n      \"method\": \"Antisense oligonucleotide knockdown in mouse bone marrow cultures; conditioned media from ADAM8-transfected cells; truncation/domain mapping constructs; pit formation assay\",\n      \"journal\": \"Journal of bone and mineral research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain mapping by truncation constructs plus functional loss-of-function, single lab\",\n      \"pmids\": [\"11341326\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"ADAM8 activation requires a novel pre-processing step: before terminal autocatalytic prodomain removal, the prodomain is first cleaved at Glu158 (N-terminal sequencing data), fracturing the pro-segment prior to removal of the cysteine switch (Cys167). This pre-processing requires the disintegrin (DIS) and cysteine-rich/EGF (CR/EGF) domains — ADAM8 constructs lacking these domains cannot complete pre-processing. Without pre-processing, the pro-segment re-associates with the catalytic domain and strongly inhibits activity.\",\n      \"method\": \"N-terminal sequencing of activation intermediates; enterokinase-cleavable construct engineering to bypass pre-processing; domain-deletion constructs; specific activity measurements\",\n      \"journal\": \"Bioscience reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — N-terminal sequencing of cleavage products plus engineered constructs and domain deletions in one study\",\n      \"pmids\": [\"18811590\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"N-glycosylation at four sites (Asn-67, Asn-91, Asn-436, Asn-612) is required for correct ADAM8 processing, localization, stability, and activity. Asn-91 glycosylation (high mannose, prodomain) is essential for exit from the ER; Asn-612 (complex type, remnant form) for exit from the Golgi and cell-surface localization; Asn-436 mutation leads to enhanced lysosomal degradation. ADAM8 dimerization on the cell surface was detected specifically in ERα-negative (but not ERα-positive) breast cancer cells and required N-glycosylation.\",\n      \"method\": \"Site-directed mutagenesis of four Asn sites; glycosidase treatment; subcellular fractionation; Western blot for processing; flow cytometry for surface localization; glycan analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — mutagenesis of all four sites with orthogonal localization and processing assays in one study\",\n      \"pmids\": [\"25336660\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"ADAM8 requires multimerization for biological function and associates with β1 integrin on the cell surface of pancreatic cancer (PDAC) cells. A peptidomimetic inhibitor (BK-1361), designed by structural modelling of the disintegrin domain, prevents ADAM8 multimerization and reduces PDAC cell invasiveness, ERK1/2 activation, and MMP activity. In vivo, BK-1361 decreased tumor burden and metastasis in mouse models of PDAC.\",\n      \"method\": \"Co-immunoprecipitation (ADAM8 with β1 integrin); structural modelling of disintegrin domain for inhibitor design; invasion assays; ERK1/2 and MMP activity assays; mouse xenograft and Kras(G12D) PDAC model\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP, cell-based functional assays, pharmacological inhibition, and in vivo validation across multiple models\",\n      \"pmids\": [\"25629724\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"In triple-negative breast cancer (TNBC), ADAM8 promotes angiogenesis through release of VEGF-A and transendothelial cell migration via β1-integrin activation. ADAM8 knockdown tumors in an orthotopic mouse model failed to grow beyond palpable size and showed poor vascularization, reduced circulating tumor cells, and reduced brain metastases. Anti-ADAM8 antibody treatment reduced primary tumor burden and metastases in a resection model.\",\n      \"method\": \"shRNA knockdown; orthotopic mouse model; antibody treatment in vivo; VEGF-A release assay; β1-integrin activation assay; transendothelial migration assay\",\n      \"journal\": \"EMBO molecular medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function with defined molecular readouts, in vivo validation, and pharmacological rescue, replicated with orthogonal approaches\",\n      \"pmids\": [\"24375628\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"ADAM8 mediates temozolomide (TMZ) chemoresistance in glioblastoma (GBM) cells by enhancing pAkt/PI3K and pERK1/2 signaling. Soluble HGF receptor/c-met and CD44 were identified as metalloprotease substrates in TMZ-treated GBM cells by proteomics; blocking HGF R/c-met prevented TMZ-induced invasiveness. ADAM8 knockdown or pharmacological inhibition (with BB-94 but not the ADAM8-sparing BB-2516) increased TMZ sensitivity.\",\n      \"method\": \"ADAM8 shRNA knockdown, overexpression, and pharmacological inhibition; proteomics substrate identification; blocking antibodies for c-met; pAkt/pERK immunoblotting; Matrigel invasion assay\",\n      \"journal\": \"Neuro-oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple approaches (KD, OE, pharmacological inhibition, proteomics substrates) in single lab\",\n      \"pmids\": [\"25825051\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ADAM8 cytoplasmic domain is required for MMP-9 upregulation: re-expression of WT ADAM8 (but not ADAM8 lacking the cytoplasmic domain) in ADAM8-knockdown breast cancer cells restored elevated pERK1/2, pCREB(S133), and MMP-9 transcription. ADAM8 also sheds PSGL-1 from breast cancer cells, and antibodies against PSGL-1 reduced transendothelial migration, linking ADAM8-mediated PSGL-1 shedding to transendothelial migration.\",\n      \"method\": \"Re-expression of WT vs. cytoplasmic-domain-deleted ADAM8; pERK/pCREB immunoblotting; qPCR for MMP-9; anti-PSGL-1 antibody blocking in transendothelial migration assay\",\n      \"journal\": \"International journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain-deletion rescue experiment plus antibody blocking, single lab\",\n      \"pmids\": [\"28986926\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"ADAM8 deficiency in mice completely prevents development of ovalbumin-induced airway inflammation and hyperresponsiveness. ADAM8 expression is required in both hematopoietic cells (including T cells) and non-hematopoietic cells for full asthma manifestation, demonstrated by bone marrow chimera experiments. Loss of ADAM8 impaired migration of T cells, eosinophils, and myeloid cells from blood vessels to the lung.\",\n      \"method\": \"Adam8-/- knockout mice; OVA-induced asthma model; bone marrow chimera experiments; bronchoalveolar lavage analysis; histology; airway hyperresponsiveness measurement\",\n      \"journal\": \"American journal of respiratory and critical care medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — knockout and chimera experiments with defined cellular and functional phenotype, replicated across multiple model conditions\",\n      \"pmids\": [\"20194813\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"ADAM8 limits allergic airway inflammation (AAI) by activating the intrinsic apoptosis pathway in myeloid leukocytes (eosinophils and macrophages): Adam8-/- mice have fewer apoptotic eosinophils and macrophages in airways during AAI, and Adam8-/- macrophages and eosinophils show reduced apoptosis when the intrinsic (but not the extrinsic) apoptosis pathway is triggered in vitro. Leukocyte-derived ADAM8 predominantly mediates this anti-inflammatory activity, confirmed by bone marrow chimeras.\",\n      \"method\": \"Adam8-/- knockout mice; bone marrow chimera; OVA and house dust mite AAI models; apoptosis assays (intrinsic vs. extrinsic pathway activation) in isolated cells; airway leukocyte counting\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO mice, chimera experiments, and cell-specific in vitro apoptosis assays with pathway discrimination\",\n      \"pmids\": [\"23670189\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"ADAM8-deficient (Adam8-/-) mice develop normally without major defects in any tissue or organ, indicating ADAM8 is dispensable for normal development and homeostasis. Expression analysis showed ADAM8 is prominently expressed during mouse development in decidua, trophoblast derivatives, gonadal ridge, thymus, cartilage/bone, brain, spinal cord, and perivenous mesenchyme.\",\n      \"method\": \"Targeted gene deletion (Adam8-/- mice); developmental expression analysis by in situ hybridization/immunostaining; histological phenotyping\",\n      \"journal\": \"Developmental dynamics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — comprehensive knockout phenotyping and expression mapping, single lab\",\n      \"pmids\": [\"15580619\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"ADAM8-mediated shedding of VCAM-1 from endothelial cells reduces eosinophil adhesion (via α4β1 integrin) and suppresses experimental asthma: ADAM8-transgenic mice (expressing ectodomain of CD156/ADAM8) show markedly reduced cellular infiltrates in peribronchovascular lesions compared to controls, associated with increased VCAM-1 shedding.\",\n      \"method\": \"ADAM8 transgenic mice; OVA-induced asthma model; VCAM-1 shedding assay in human umbilical vein endothelial cells stimulated with ADAM8-transgenic cells\",\n      \"journal\": \"Immunology letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — transgenic mouse model with defined substrate (VCAM-1 shedding) and in vitro endothelial cell assay, single lab\",\n      \"pmids\": [\"16154205\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"ADAM8 overexpression in osteoclast (OCL) precursors (TRAP-ADAM8 transgenic mice) produces osteopenia, hypermultinucleated OCLs with increased bone resorption capacity, enhanced OCL precursor fusion, increased TRAF6 expression, and elevated NF-κB, Erk, and Akt signaling, as well as increased p-Pyk2 and p-Src activation. ADAM8 knockout mice do not display a basal bone phenotype but fail to increase RANKL production, OCL formation, or calvarial fibrosis in response to TNF-α.\",\n      \"method\": \"TRAP-ADAM8 transgenic overexpression mice; Adam8-/- knockout mice; TNF-α calvarial injection model; osteoclast formation and bone resorption assays; Western blot for TRAF6, NF-κB, Erk, Akt, p-Pyk2, p-Src\",\n      \"journal\": \"Journal of bone and mineral research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — complementary gain- and loss-of-function in vivo models with defined signaling pathway readouts\",\n      \"pmids\": [\"20683884\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Catalytically inactive ADAM8 (E330Q knock-in transgenic DBA/1J mice) shows decreased incidence and severity of collagen-induced arthritis with reduced synovial inflammation, cartilage degradation, and bone resorption, demonstrating that ADAM8 proteolytic activity is required for experimental arthritis development.\",\n      \"method\": \"Transgenic mice expressing E330Q catalytically inactive ADAM8; LPS-synchronized collagen-induced arthritis model; histological scoring of joints; expression profiling\",\n      \"journal\": \"Clinical and experimental immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — catalytically inactive knock-in with in vivo disease phenotype, single lab but rigorous genetic approach\",\n      \"pmids\": [\"19737139\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"ADAM8 overexpression in endothelial cells increases ectodomain shedding of multiple pro-angiogenic membrane proteins including CD31, Tie-2, Flk-1, Flt-1, EphrinB2, EphB4, VE-cadherin, KL-1, E-selectin, and neuregulin-1β2. In Adam8-/- mice, the oxygen-induced retinopathy model shows more retinal re-vascularization but fewer neovascular tufts, and heterotopically injected tumor cells grow faster, suggesting ADAM8 limits pathological angiogenesis.\",\n      \"method\": \"ADAM8 overexpression in endothelial cells with ectodomain shedding assays; Adam8-/- mice in OIR model; tumor cell heterotopic injection model\",\n      \"journal\": \"Journal of molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell overexpression shedding assay plus knockout in vivo models, single lab\",\n      \"pmids\": [\"20119708\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ADAM8 promotes migration of IL-5-stimulated eosinophils on periostin-rich extracellular matrix: ADAM8 on eosinophils degrades (sheds the Stiny-1 epitope of) periostin in the extracellular matrix and is required for migration. Shed ADAM8 proteoforms lacking the cytoplasmic tail were detected in supernatants. ADAM8 antibodies and metalloproteinase inhibitors both blocked eosinophil migration and periostin epitope clearance.\",\n      \"method\": \"Fluorescence microscopy of eosinophil migration on periostin; anti-ADAM8 and metalloprotease inhibitor blocking; Western blot of ADAM8 proteoforms in supernatants\",\n      \"journal\": \"Clinical and experimental allergy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — antibody blocking with defined substrate and functional readout, single lab\",\n      \"pmids\": [\"28378503\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Leukocytic ADAM8 is required for chemokine-induced chemotaxis and transendothelial and transepithelial migration of monocytic THP-1 cells, as well as αL-integrin upregulation and THP-1 adhesion to endothelial cells. On endothelial cells, ADAM8 enhances transendothelial migration and cytokine-induced permeability. In Adam8-/- mice, intranasal LPS-induced acute lung inflammation shows reduced leukocyte infiltration. Bone marrow macrophage transfer experiments confirmed a predominantly leukocyte-intrinsic promigratory function.\",\n      \"method\": \"shRNA knockdown in THP-1 cells; Adam8-/- knockout mice; LPS-induced acute lung inflammation model; BMDM transfer experiments; transendothelial/transepithelial migration assays; αL-integrin flow cytometry\",\n      \"journal\": \"American journal of physiology. Lung cellular and molecular physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — knockdown, knockout mouse, and adoptive transfer with defined functional phenotype across orthogonal assay systems\",\n      \"pmids\": [\"28596294\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ADAM8 regulates angiogenesis in glioblastoma via the JAK/STAT3 pathway controlling osteopontin (OPN) expression: ADAM8 knockdown in U87 glioma cells decreased angiogenesis and tumor volumes in vivo. In vitro, ADAM8 regulates OPN through JAK/STAT3 signaling in both glioma cells and primary macrophages.\",\n      \"method\": \"ADAM8 shRNA knockdown; stereotactic mouse brain injection model; in vitro angiogenesis assays; JAK/STAT3 pathway inhibition; OPN measurement\",\n      \"journal\": \"Biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockdown with in vivo validation and defined signaling pathway, single lab\",\n      \"pmids\": [\"33544472\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ADAM8 is present as an active protease in extracellular vesicles (EVs) from PDAC cells and is associated with lipocalin 2 (LCN2) and MMP-9 in an ADAM8-dependent manner (ADAM8 KO cells show reduced LCN2 and MMP-9 in EVs). ADAM8 sorting into EVs is independent of the TSG101 ESCRT-I pathway despite the presence of a PTAP recognition motif in ADAM8's cytoplasmic domain.\",\n      \"method\": \"ADAM8 CRISPR KO in PDAC cells; EV isolation and proteomics; Western blot for LCN2, MMP-9, TSG101 in EVs; co-culture with macrophages\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO with EV proteomics and functional co-culture, single lab\",\n      \"pmids\": [\"35216088\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In macrophages after myocardial infarction, ADAM8 binds to ANXA2 and promotes phosphorylation of ANXA2 at Ser26. ADAM8 knockout impedes ANXA2 phosphorylation, inhibits mTOR Ser2448 phosphorylation, and activates autophagy, leading to enhanced angiogenesis and suppressed inflammation. Macrophage-specific ADAM8 KO mice show improved cardiac repair, confirmed by bone marrow transplantation; these effects are reversed by ADAM8 overexpression.\",\n      \"method\": \"Macrophage-specific ADAM8 KO (CRISPR/Cas9, Lyz2-Cre); bone marrow transplantation; Co-IP/mass spectrometry (ADAM8-ANXA2 interaction); RNA sequencing; phospho-specific Western blots (p-mTOR, p-ANXA2); ANXA2 phosphorylation activation/inactivation experiments\",\n      \"journal\": \"Journal of advanced research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — Co-IP/MS interaction data plus functional rescue experiments and genetic confirmation, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"39097092\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"Human CD156 (ADAM8) cDNA encodes a transmembrane glycoprotein with an extracellular region containing metalloprotease, disintegrin, cysteine-rich, and EGF-like domains homologous to hemorrhagic snake venom proteins. mRNA is expressed in macrophage cell lines, granulocytes, monocytes, and B cells but not T cell lines. The gene maps to human chromosome 10q26.3.\",\n      \"method\": \"cDNA cloning and sequencing from THP-1 macrophage and granulocyte libraries; Northern blot expression analysis; chromosomal mapping\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — cDNA sequencing and expression analysis establishing domain architecture and cell-type specificity, replicated across labs\",\n      \"pmids\": [\"9126482\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"The murine CD156/ADAM8 gene spans ~14 kb, is composed of 24 exons, and encodes a protein with metalloprotease domain (exons 7–12), disintegrin domain (exons 12–15), and transmembrane region (single exon 19). The promoter contains cis-acting elements at positions −183, −334, and −623 and LPS-inducible elements at −183 and −390, and interferon-γ-inducible regions at −202, −507, and −659, defined by CAT reporter assays in monocytic cells.\",\n      \"method\": \"Genomic cloning, sequencing and exon mapping; chloramphenicol acetyltransferase (CAT) reporter assays with promoter deletion constructs in monocytic cell line; LPS and IFN-γ stimulation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — promoter deletion analysis with reporter assays and stimulation experiments, single lab\",\n      \"pmids\": [\"9218457\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"ADAM8 inhibition by hydroxamate-based inhibitors: BB-94 (batimastat), GW280264, FC387, and FC143 inhibit ADAM8 in vitro and in cell-based CD23 shedding assays; GM6001, TAPI2, and BB-2516 (marimastat) weakly inhibit ADAM8; GI254023 (ADAM10-specific) does not inhibit ADAM8. Structural modelling of the S1 position reveals T300 in ADAM8 (and T347 in ADAM17) vs. V327 in ADAM10 explains differential inhibitor selectivity.\",\n      \"method\": \"In vitro recombinant ADAM8 protease assay; cell-based CD23 shedding assay in HEK293 cells; structural modelling of inhibitor binding\",\n      \"journal\": \"Biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro enzymatic assay plus cell-based validation and structural modelling, single lab\",\n      \"pmids\": [\"30738011\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In glioblastoma, ADAM8 activates HB-EGF/EGFR signaling to upregulate CCL2 expression in tumor cells, which recruits tumor-associated macrophages (TAMs). TAMs in turn induce ADAM8 expression in GBM cells, forming a positive feedback loop that mediates TMZ chemoresistance. Only ADAM8 (and not other ADAM or MMP family members screened) was upregulated in GBM cells by macrophages under TMZ treatment.\",\n      \"method\": \"qPCR screen of ADAM/MMP family in co-culture; RNA-seq; ELISA and Western blot for CCL2; EGFR signaling analysis; in vitro and in vivo TAM recruitment assays; TCGA data analysis with IHC validation\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic co-culture screen plus pathway analysis and in vivo validation, single lab\",\n      \"pmids\": [\"36230833\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ADAM8 is a membrane-anchored metalloprotease-disintegrin that undergoes obligatory autocatalytic prodomain removal (requiring a DIS/CR/EGF domain-coordinated pre-processing step at Glu158 before final cysteine-switch removal at Cys167, with N-glycosylation at Asn-91 and Asn-612 essential for correct ER/Golgi exit and cell-surface localization), after which it acts as a sheddase for multiple substrates including CD23, L-selectin, VCAM-1, CHL1, APP, PSGL-1, and angiogenic receptors; its disintegrin domain mediates cell adhesion and, through multimerization, associates with β1 integrin to activate ERK1/2, Akt/PI3K, and FAK signaling, while its cytoplasmic domain drives MMP-9 transcription via pERK/pCREB and in macrophages binds ANXA2 to regulate mTOR-autophagy, collectively enabling ADAM8 to control leukocyte migration, osteoclast differentiation and bone resorption, airway inflammation (with both pro- and anti-inflammatory roles depending on cell type), and cancer cell invasiveness, angiogenesis, and chemoresistance.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ADAM8 (CD156) is a membrane-anchored metalloprotease-disintegrin that couples regulated ectodomain shedding to adhesion-dependent signaling, controlling leukocyte migration, osteoclast-mediated bone resorption, angiogenesis, and tumor invasion [#13, #17, #10, #25]. Its protease domain is activated by an obligatory autocatalytic mechanism: a pre-processing cleavage at Glu158 that requires the disintegrin and cysteine-rich/EGF domains fractures the prodomain before final cysteine-switch removal at Cys167, and active-site mutation (E330Q) of the HExxH motif abolishes activity [#0, #7]. Correct maturation also depends on site-specific N-glycosylation, with Asn-91 required for ER exit and Asn-612 for Golgi exit and cell-surface display [#8]. Unusually among ADAMs, ADAM8 is resistant to all four TIMPs but inhibited by hydroxamate-based compounds, a selectivity traced to an S1-pocket threonine (T300) [#1, #27]. Once active, ADAM8 sheds a broad substrate set including CD23, L-selectin, CHL1, APP, VCAM-1, PSGL-1, periostin, and pro-angiogenic receptors, generating products with distinct downstream consequences [#2, #3, #5, #16, #20, #19]. Beyond catalysis, the disintegrin domain mediates cell adhesion and, through multimerization, associates with β1 integrin to drive ERK1/2, Akt/PI3K, and downstream invasion programs, while the cytoplasmic domain is required for pERK/pCREB-dependent MMP-9 transcription [#0, #9, #10, #12]. In disease contexts ADAM8 promotes pancreatic, breast, and glioblastoma tumor invasion, angiogenesis, and chemoresistance [#9, #10, #11], and is genetically required for allergic airway inflammation, arthritis, and inflammatory osteoclastogenesis, though it can also exert anti-inflammatory effects by promoting intrinsic apoptosis of myeloid leukocytes and, in macrophages, by binding ANXA2 to regulate mTOR-autophagy [#13, #18, #17, #14, #24].\"\n  ,\n  \"teleology\": [\n    {\n      \"year\": 1997,\n      \"claim\": \"Establishing ADAM8's domain architecture and expression pattern defined it as a candidate metalloprotease-disintegrin of myeloid lineage, framing all later mechanistic work.\",\n      \"evidence\": \"cDNA cloning/sequencing, Northern blot, and chromosomal mapping in human; genomic exon mapping and promoter reporter assays in mouse\",\n      \"pmids\": [\"9126482\", \"9218457\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not demonstrate catalytic activity or identify substrates\", \"Promoter elements mapped only in monocytic cells\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Mapping osteoclast-stimulatory activity to the disintegrin/cysteine-rich domain showed ADAM8 has non-proteolytic adhesion/signaling functions in bone biology.\",\n      \"evidence\": \"Antisense knockdown, secretable-form conditioned media, and truncation constructs in mouse bone marrow cultures with pit formation assay\",\n      \"pmids\": [\"11341326\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor/ligand partner for the disintegrin domain not identified\", \"Single-lab loss-of-function\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Reconstitution with active-site mutants established ADAM8 as a genuine autocatalytically activated metalloprotease distinct from TIMP-regulated ADAMs/MMPs.\",\n      \"evidence\": \"E330Q active-site mutagenesis, co-transfection trans-cleavage rescue, in vitro protease assays with purified enzyme, and TIMP/hydroxamate inhibition tests\",\n      \"pmids\": [\"12372841\", \"12135759\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological substrates not yet defined\", \"Structural basis of TIMP resistance unresolved at this stage\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Identification of CD23 as a shed substrate connected ADAM8 catalysis to a defined immune-relevant cleavage event.\",\n      \"evidence\": \"Soluble CD23 release with active vs. E330Q ADAM8, co-IP of ADAM8 with membrane CD23, and peptide library specificity profiling\",\n      \"pmids\": [\"12777399\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo CD23 shedding by ADAM8 not demonstrated\", \"Recognition determinants only partially mapped\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"CHL1 was established as a specific ADAM8 substrate with a functional neural readout, distinguishing ADAM8 from ADAM10/17.\",\n      \"evidence\": \"In vitro CHL1-Fc cleavage, WT vs. E330Q transfection, Adam8-/- brain extracts, and co-culture neurite/apoptosis assays\",\n      \"pmids\": [\"14761956\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological neural phenotype of impaired CHL1 processing not characterized in vivo\", \"Receptor mediating processed-CHL1 effects unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Profiling substrate peptides and APP cleavage broadened the substrate repertoire and defined sequence criteria for ADAM8 cleavage.\",\n      \"evidence\": \"ProteaseSpot peptide cleavage with recombinant ADAM8, APP cleavage in HEK293 with WT vs. inactive enzyme, and cleavage-site mutagenesis\",\n      \"pmids\": [\"16542157\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Many peptide hits not validated as full-length substrates in vivo\", \"Single-lab study\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Neutrophil studies showed ADAM8 is granule-stored, activation-mobilized to the surface, and itself shed, linking its trafficking to leukocyte function.\",\n      \"evidence\": \"Subcellular fractionation, confocal imaging of granule mobilization, and L-selectin shedding assay\",\n      \"pmids\": [\"17548643\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism triggering granule mobilization not defined\", \"L-selectin shedding shown in heterologous cells\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defining a DIS/CR-EGF-dependent pre-processing step at Glu158 prior to Cys167 removal explained how ADAM8 escapes prodomain auto-inhibition.\",\n      \"evidence\": \"N-terminal sequencing of intermediates, enterokinase-bypass constructs, and domain-deletion specific-activity measurements\",\n      \"pmids\": [\"18811590\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether pre-processing is intra- or intermolecular not fully resolved\", \"Trafficking compartment of each step not localized\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Knockout and transgenic mouse models established ADAM8 as dispensable for development but functionally required and substrate-directed (VCAM-1) in inflammatory contexts.\",\n      \"evidence\": \"Adam8-/- phenotyping with developmental expression mapping; ADAM8-transgenic mice in OVA asthma with VCAM-1 shedding assays\",\n      \"pmids\": [\"15580619\", \"16154205\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional redundancy masking developmental roles not excluded\", \"VCAM-1 shedding study single-lab and transgenic-driven\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Genetic and chimera experiments placed ADAM8 as required for allergic airway inflammation through promigratory roles in both hematopoietic and non-hematopoietic compartments, while also limiting pathological angiogenesis.\",\n      \"evidence\": \"Adam8-/- mice in OVA asthma with bone marrow chimeras; ADAM8 overexpression shedding assays plus Adam8-/- OIR and tumor models\",\n      \"pmids\": [\"20194813\", \"20119708\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Causal substrate(s) mediating the migration defect not pinpointed\", \"Pro- vs anti-angiogenic balance context-dependence unresolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Complementary gain- and loss-of-function osteoclast models linked ADAM8 to TRAF6/NF-κB/Erk/Akt signaling and inflammatory bone resorption.\",\n      \"evidence\": \"TRAP-ADAM8 transgenic and Adam8-/- mice with TNF-α calvarial model and pathway immunoblots\",\n      \"pmids\": [\"20683884\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct molecular link from ADAM8 to TRAF6 induction not defined\", \"Catalytic vs. adhesion contribution to osteoclast phenotype not dissected\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"A catalytically inactive E330Q knock-in established that ADAM8 proteolytic activity is required for experimental arthritis.\",\n      \"evidence\": \"E330Q knock-in DBA/1J mice in collagen-induced arthritis with histological scoring\",\n      \"pmids\": [\"19737139\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relevant arthritic substrates not identified\", \"Single-lab disease model\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Studies in TNBC and AAI revealed a dual role: ADAM8 drives β1-integrin-dependent angiogenesis and metastasis in cancer yet limits airway inflammation by promoting intrinsic apoptosis of myeloid leukocytes.\",\n      \"evidence\": \"shRNA knockdown, orthotopic and resection mouse models, anti-ADAM8 antibody therapy, VEGF-A/transendothelial assays; Adam8-/- mice with chimeras and intrinsic vs extrinsic apoptosis assays\",\n      \"pmids\": [\"24375628\", \"23670189\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of cell-type-dependent pro- vs anti-inflammatory switch unresolved\", \"Apoptosis-promoting mechanism downstream of ADAM8 not defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Site-specific N-glycosylation was shown to control ADAM8 ER/Golgi trafficking, stability, and surface dimerization, with dimerization selective to ERα-negative breast cancer cells.\",\n      \"evidence\": \"Mutagenesis of four Asn sites, glycosidase treatment, fractionation, flow cytometry, and glycan analysis\",\n      \"pmids\": [\"25336660\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Why dimerization is restricted to ERα-negative cells not mechanistically explained\", \"Glycan-dependent partner interactions not mapped\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Multimerization-dependent association with β1 integrin was shown to drive cancer invasion and was therapeutically targetable, while ADAM8 was implicated in glioblastoma chemoresistance via pAkt/pERK signaling.\",\n      \"evidence\": \"Co-IP with β1 integrin, structure-guided peptidomimetic BK-1361, invasion/ERK/MMP assays and PDAC mouse models; GBM knockdown/overexpression/inhibition with proteomic substrate ID (c-met, CD44)\",\n      \"pmids\": [\"25629724\", \"25825051\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether β1-integrin signaling requires catalytic activity not fully separated\", \"GBM substrate-to-resistance causal chain partly correlative\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Cytoplasmic-domain and leukocyte-migration studies dissected adhesion/signaling functions: the cytoplasmic tail drives pERK/pCREB-dependent MMP-9 transcription and PSGL-1 shedding, and leukocytic ADAM8 is intrinsically required for chemotaxis and transmigration.\",\n      \"evidence\": \"WT vs. cytoplasmic-deletion rescue, qPCR/immunoblot, anti-PSGL-1 blocking; THP-1 knockdown, Adam8-/- LPS lung model, BMDM transfer, and αL-integrin/migration assays; eosinophil migration on periostin with ADAM8/protease blocking\",\n      \"pmids\": [\"28986926\", \"28596294\", \"28378503\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct effectors binding the cytoplasmic tail to drive ERK/CREB not all identified\", \"Relative weight of shedding vs. integrin signaling in migration unresolved\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Tumor microenvironment and vesicle studies extended ADAM8's reach: it forms an HB-EGF/EGFR–CCL2–TAM feedback loop in glioblastoma and is packaged with LCN2/MMP-9 into extracellular vesicles via a TSG101-independent route.\",\n      \"evidence\": \"Co-culture qPCR screen, RNA-seq, CCL2/EGFR analyses, TAM recruitment models; CRISPR KO PDAC cells with EV proteomics and macrophage co-culture\",\n      \"pmids\": [\"36230833\", \"35216088\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"EV sorting mechanism replacing ESCRT-I not identified\", \"In vivo relevance of EV-borne ADAM8 not established\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"A non-proteolytic intracellular function emerged: macrophage ADAM8 binds and phosphorylates ANXA2 to sustain mTOR signaling and suppress autophagy, impairing post-infarct cardiac repair.\",\n      \"evidence\": \"Macrophage-specific ADAM8 KO, bone marrow transplantation, Co-IP/MS, RNA-seq, and phospho-specific immunoblots (p-ANXA2, p-mTOR)\",\n      \"pmids\": [\"39097092\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a membrane sheddase accesses cytoplasmic ANXA2 to drive its phosphorylation not mechanistically explained\", \"Single-lab study\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How ADAM8 selects between its proteolytic (sheddase), adhesive (disintegrin/β1-integrin), and intracellular (cytoplasmic-tail/ANXA2) modes in a cell-type-specific manner — and how this dictates its opposing pro- versus anti-inflammatory and pro- versus anti-angiogenic outcomes — remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No high-resolution structure of full-length or multimerized ADAM8\", \"Unifying model integrating catalysis, integrin signaling, and intracellular partners absent\", \"Determinants of context-dependent functional polarity unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 2, 3, 4, 5]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 1, 27]},\n      {\"term_id\": \"GO:0098631\", \"supporting_discovery_ids\": [0, 6, 9]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [9, 10, 24]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [5, 8, 9]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [5, 23]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [13, 14, 21]},\n      {\"term_id\": \"R-HSA-1474244\", \"supporting_discovery_ids\": [3, 20]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [9, 17, 24]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [9, 10, 11, 28]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 7, 8]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"ITGB1\", \"ANXA2\", \"CD23\", \"CHL1\", \"PSGL1\", \"LCN2\", \"MMP9\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":7,"faith_total":7,"faith_pct":100.0}}