{"gene":"HBEGF","run_date":"2026-06-10T01:55:21","timeline":{"discoveries":[{"year":1995,"finding":"Membrane-anchored proHB-EGF forms a complex with the tetraspanin DRAP27/CD9 and integrin α3β1 at cell-cell contact sites, as demonstrated by co-immunoprecipitation, chemical cross-linking, and double-immunofluorescence colocalization in Vero, A431, and MG63 cells.","method":"Co-immunoprecipitation, chemical cross-linking, double-immunofluorescence microscopy","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP confirmed by chemical cross-linking and colocalization, multiple cell lines tested","pmids":["7790364"],"is_preprint":false},{"year":1997,"finding":"CD9, but not CD81 or CD82, upregulates the juxtacrine mitogenic activity of transmembrane proHB-EGF as a diphtheria toxin receptor; chimeric CD9/CD81 molecules mapped this activity to the second half of CD9 (large extracellular loop, fourth TM domain, and C-terminal cytoplasmic domain).","method":"Cotransfection of chimeric CD9/CD81 constructs in murine LM cells, diphtheria toxin receptor functional assay","journal":"Cellular immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain-mapping by chimeric constructs with functional readout, single lab","pmids":["9514697"],"is_preprint":false},{"year":2000,"finding":"Estrogen-induced ERK1/2 activation requires GPR30-dependent transactivation of the EGF receptor via release of HB-EGF; this was blocked by neutralizing HB-EGF antibodies or by down-modulating HB-EGF from the cell surface with the diphtheria toxin mutant CRM-197, and required Src-related tyrosine kinase activity and Shc-pY317 phosphorylation.","method":"Neutralizing antibody treatment, CRM-197 cell-surface depletion, EGFR kinase inhibition, GPR30 cDNA transfection, immunoblot for pERK","journal":"Molecular endocrinology (Baltimore, Md.)","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal inhibitors (antibody, CRM-197, kinase inhibitor, dominant-negative transfection) converging on same conclusion","pmids":["11043579"],"is_preprint":false},{"year":2002,"finding":"ADAM12 is the metalloprotease responsible for shedding HB-EGF in cardiomyocytes in response to GPCR agonists; dominant-negative ADAM12 expression abrogated shedding-dependent EGFR transactivation and cardiac hypertrophy, and the metalloprotease inhibitor KB-R7785 bound directly to ADAM12.","method":"ADAM12 cloning, dominant-negative expression, metalloprotease inhibitor (KB-R7785) direct binding, in vivo cardiac hypertrophy model","journal":"Nature medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — dominant-negative genetic approach plus pharmacological inhibitor with direct binding data, replicated in vivo","pmids":["11786904"],"is_preprint":false},{"year":2002,"finding":"HB-EGF shedding via metalloprotease (MMP) and subsequent EGFR transactivation is required for Angiotensin II-induced fibronectin expression in mesangial cells; ALP-tagged HB-EGF chimera directly measured rapid HB-EGF release (~4-fold within 2 min of Ang II), blocked by PKC inhibitors or MMP inhibitor batimastat.","method":"HB-EGF-alkaline phosphatase chimera shedding assay, neutralizing anti-HB-EGF antibody, EGFR inhibitor AG1478, heparin blocking, batimastat MMP inhibition","journal":"Kidney international","confidence":"High","confidence_rationale":"Tier 1 / Moderate — direct measurement of shedding with reporter chimera, multiple pharmacological blockers, single lab","pmids":["11737589"],"is_preprint":false},{"year":2002,"finding":"Helicobacter pylori triggers EGFR tyrosine phosphorylation via HB-EGF gene induction and metalloprotease-dependent ectodomain shedding (the 'triple membrane passing signal'); this signaling also enhances IL-8 secretion.","method":"EGFR phosphorylation assay, HB-EGF gene expression analysis, metalloprotease inhibition, EGFR and MEK1 inhibitors","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple pathway inhibitors, pharmacological validation, single lab","pmids":["12099696"],"is_preprint":false},{"year":2002,"finding":"TGF-β mediates fibronectin expression in mesangial cells via PC-PLC/PKC-dependent metalloprotease-mediated HB-EGF release and subsequent EGFR transactivation; Smad2 phosphorylation by TGF-β was unaffected by EGFR inhibition, and HB-EGF did not activate Smad2, indicating a parallel pathway.","method":"HB-EGF-alkaline phosphatase chimera shedding assay, neutralizing anti-HB-EGF antibody, AG1478, PKC inhibitors, batimastat, FN mRNA quantification","journal":"Kidney international","confidence":"High","confidence_rationale":"Tier 1 / Moderate — direct shedding reporter, multiple inhibitors, epistasis between Smad2 and EGFR pathways established, single lab","pmids":["12164862"],"is_preprint":false},{"year":2003,"finding":"HB-EGF null mice exhibit enlarged, malformed cardiac valves due to abnormal mesenchymal cell proliferation during remodeling, associated with increased activated Smad1/5/8; similar defects occur in EGFR- and TACE-null mice, establishing that TACE-derived soluble HB-EGF activates EGFR to suppress BMP signaling during cardiac valvulogenesis.","method":"Homologous recombination knockout mice, genetic epistasis (HB-EGF−/−, EGFR−/−, TACE−/−), immunohistochemistry for pSmad1/5/8, histological analysis","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — three independent genetic knockouts show convergent cardiac valve phenotype, epistasis established","pmids":["12773386"],"is_preprint":false},{"year":2004,"finding":"Wound-induced MMP-dependent ectodomain shedding of HB-EGF acts as an autocrine/paracrine EGFR ligand in corneal epithelial wound closure; wound-induced EGFR and ERK phosphorylation were blocked by CRM-197 (HB-EGF antagonist), HB-EGF function-blocking antibodies, and MMP inhibitor GM6001, with direct HB-EGF shedding demonstrated by HB-EGF-AP reporter.","method":"HB-EGF-alkaline phosphatase shedding assay, CRM-197, neutralizing HB-EGF antibodies, GM6001 MMP inhibitor, AG1478 EGFR inhibitor, immunoprecipitation + Western blot","journal":"Investigative ophthalmology & visual science","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct shedding reporter plus multiple orthogonal inhibitors confirmed in both organ culture and cell culture models","pmids":["14985295"],"is_preprint":false},{"year":2004,"finding":"Soluble HB-EGF, but not pro-HB-EGF (membrane-only form), confers tumorigenic properties including enhanced growth, colony formation, cell migration, cyclin D1 activation, VEGF induction, and MMP-9/MMP-3 upregulation in bladder cancer cells in vitro and in vivo.","method":"Tetracycline-regulatable expression system, soft agar colony assay, xenograft nude mouse tumorigenesis, zymography for MMP activity","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — inducible expression with paired s-HB-EGF vs pro-HB-EGF controls, multiple in vitro and in vivo readouts","pmids":["15289334"],"is_preprint":false},{"year":2004,"finding":"Membrane-anchored (noncleavable) HB-EGF promotes cell-matrix and cell-cell interactions and decreases migration and HGF-induced scattering in MDCK cells, while soluble HB-EGF has the opposite effects, demonstrating that the two forms of HB-EGF have distinctly different functional consequences.","method":"Stable transfection of proHB-EGF, noncleavable deletion mutant, or soluble HB-EGF in MDCK cells; migration, scattering, tubulogenesis, and adhesion assays","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic approach with matched cell lines expressing distinct forms, multiple orthogonal functional assays, single lab","pmids":["14996914"],"is_preprint":false},{"year":2004,"finding":"HB-EGF directs stromal cell polyploidy and decidualization via upregulation of cyclin D3; adenoviral antisense delivery of cyclin D3 abrogated HB-EGF-induced polyploidy both in vitro and in vivo.","method":"Adenoviral antisense cyclin D3 delivery, BrdU labeling, flow cytometry, in vivo uterine decidualization model","journal":"Developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function via antisense in vitro and in vivo with specific phenotypic readout, single lab","pmids":["14697362"],"is_preprint":false},{"year":2005,"finding":"HB-EGF promotes eyelid closure by activating EGFR-ERK signaling at the leading edge of migrating epithelium; HB-EGF null and secretion-deficient mutant mice show delayed eyelid closure with reduced actin bundle formation; genetic interaction with hypomorphic EGFR (waved-2) confirmed; TGFα acts synergistically and equally.","method":"HB-EGF null mouse KO, secretion-deficient knock-in (HB-uc/uc), EGFR kinase inhibitor in vivo, double null crosses (HB-EGF × TGFα), pEGFR and pERK immunostaining","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple genetic models (KO, knock-in, hypomorph, double null) with converging mechanistic pathway","pmids":["16141218"],"is_preprint":false},{"year":2005,"finding":"HB-EGF enhances intestinal restitution via PI3K/Akt and MEK/ERK1/2 signaling downstream of ErbB-1; blocking ErbB-1, PI3K/Akt, or MEK/ERK significantly reduced intrinsic and HB-EGF-induced restitution in vitro, and endogenous HB-EGF was shown to be essential for wound-induced ErbB-1 and ERK1/2 activation.","method":"Intestinal I/R model in rats, scrape wound assay in vitro, pharmacological inhibitors of ErbB-1/PI3K/MEK, CRM-197 HB-EGF blockade, immunoblot for pAkt and pERK","journal":"Gastroenterology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo and in vitro models with pathway inhibitors and endogenous HB-EGF neutralization, single lab","pmids":["16083716"],"is_preprint":false},{"year":2006,"finding":"5-HT2A receptor induces ERK phosphorylation and cellular proliferation via TACE (ADAM17) activation, HB-EGF shedding, and EGFR transactivation in mesangial cells; TACE was co-immunoprecipitated with HB-EGF, and TACE siRNA blocked HB-EGF shedding, ERK phosphorylation, and DNA content increase.","method":"TACE siRNA knockdown, neutralizing HB-EGF antibody, EGFR inhibitor AG1478, MMP inhibitors, co-immunoprecipitation of TACE with HB-EGF, DNA content measurement","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — siRNA, pharmacological inhibitors, and Co-IP converging on TACE–HB-EGF complex; multiple orthogonal methods","pmids":["16737974"],"is_preprint":false},{"year":2006,"finding":"MMP-7 catalyzes release of HB-EGF to mediate bile acid (deoxycholyltaurine)-induced EGFR transactivation and colon cancer cell proliferation; MMP-7 and proHB-EGF co-localize at the cell surface, and specific MMP-7 knockdown (neutralizing antibody or siRNA) attenuated DCT-induced signaling and proliferation.","method":"MMP-7 siRNA, neutralizing antibody to MMP-7, CRM-197 HB-EGF inhibitor, EGFR ligand-domain antibody, immunofluorescence colocalization, qRT-PCR for MMP-7","journal":"Biochemical pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA and neutralizing antibody plus colocalization evidence, single lab","pmids":["17222808"],"is_preprint":false},{"year":2007,"finding":"HB-EGF promotes angiogenesis (migration and tube formation) in HUVEC via activation of PI3K, MAPK, and eNOS pathways, in a VEGF-independent manner.","method":"Scratch wound migration assay, 2D tube formation assay, pharmacological inhibitors of PI3K, MAPK, eNOS","journal":"Growth factors (Chur, Switzerland)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple pathway inhibitors with functional angiogenesis readouts, single lab","pmids":["18092233"],"is_preprint":false},{"year":2008,"finding":"ERK1/2 mediates ADAM-dependent HB-EGF shedding and EGFR transactivation in corneal epithelial cells in response to wounding and GPCR ligands; ADAM17 was co-immunoprecipitated with active ERK and phosphorylated at serine residues in an ERK-dependent manner.","method":"MEK inhibitors PD98059/U0126, HB-EGF-AP shedding assay, co-immunoprecipitation of ERK with ADAM17, immunoprecipitation + Western blot for serine phosphorylation of ADAM17","journal":"Investigative ophthalmology & visual science","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP of ADAM17-ERK complex plus phosphorylation evidence plus direct shedding reporter, multiple stimuli tested","pmids":["18658095"],"is_preprint":false},{"year":2008,"finding":"HB-EGF stimulates eNOS expression and NO production in endothelial cells via a PI3K-dependent pathway; eNOS siRNA and eNOS-specific inhibitors abolished HB-EGF-induced HUVEC migration and angiogenesis in vitro and in vivo (Matrigel plug assay).","method":"eNOS siRNA knockdown, L-NAME/L-NIO pharmacological eNOS inhibitors, PI3K inhibitor, Matrigel plug in vivo angiogenesis, Western blot for eNOS protein/phosphorylation","journal":"Growth factors (Chur, Switzerland)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA plus pharmacological inhibitors plus in vivo assay, single lab","pmids":["18925469"],"is_preprint":false},{"year":2009,"finding":"The heparin-binding domain of transmembrane proHB-EGF mediates localization to cell-cell contact sites via trans interaction with extracellular heparan sulfate proteoglycans (HSPGs); disruption of this interaction increases proteolytic release of soluble ligand and switches cell behavior from juxtacrine growth inhibition to autocrine proliferation.","method":"Independent tracking of extracellular EGF domain and cytoplasmic C-terminus with new labeling method, heparin treatment, HSPG disruption experiments, wound-closure assay, proliferation assays","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — novel dual-domain tracking method with functional readouts, single lab","pmids":["20530570"],"is_preprint":false},{"year":2009,"finding":"Soluble HB-EGF induces epithelial-to-mesenchymal transition (EMT) in renal collecting duct cells by upregulating the E-cadherin transcriptional repressor Snail-2; stable Snail-2 knockdown restored epithelial markers but not anchorage-independent growth.","method":"Stable overexpression of sHB-EGF, qRT-PCR and luciferase reporter for E-cadherin transcription, stable shRNA knockdown of Snail-2, Western blot for epithelial/mesenchymal markers","journal":"American journal of physiology. Renal physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain-of-function and epistasis via Snail-2 knockdown, single lab","pmids":["19244405"],"is_preprint":false},{"year":2009,"finding":"Mechanical stretch promotes fetal type II epithelial cell differentiation via ectodomain shedding of HB-EGF and TGF-α; 5% cyclic stretch of E19 fetal cells transfected with AP-tagged HB-EGF directly measured HB-EGF release, which was not enhanced by fibroblast co-culture.","method":"AP-tagged HB-EGF shedding assay under cyclic stretch, SP-B/C mRNA and protein measurement, recombinant HB-EGF/TGFα/amphiregulin/BTC/EPR comparison","journal":"The Journal of physiology","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — direct shedding reporter under defined mechanical conditions, single lab","pmids":["19237431"],"is_preprint":false},{"year":2009,"finding":"Integrin/FAK signaling masks HB-EGF's growth-promoting activity in standard 2D monolayer culture; reducing integrin β1 or FAK by antibody or genetic knockout revealed HB-EGF-dependent cell growth; HB-EGF-driven growth was clearly demonstrated in 3D culture or when integrin signaling was attenuated.","method":"Integrin β1 antibody blocking, FAK knockout, 3D culture system, xenograft tumor growth comparison","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO and antibody approaches in multiple culture contexts, single lab","pmids":["19887590"],"is_preprint":false},{"year":2011,"finding":"HB-EGF induces cardiomyocyte hypertrophy via an EGFR-MEK5-ERK5-MEF2A-COX-2 signaling pathway; MEK5 siRNA reduced HB-EGF-induced cell size and ANF mRNA expression, MEF2A siRNA attenuated COX-2 induction, and COX-2 inhibitor rofecoxib reduced ANF expression.","method":"MEK5 siRNA, MEF2A siRNA, COX-2 inhibitor rofecoxib, AG1478 EGFR inhibitor, [3H]-leucine incorporation, ANF mRNA measurement","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple siRNA knockdowns defining sequential pathway, single lab","pmids":["21244855"],"is_preprint":false},{"year":2011,"finding":"LIV-1 overexpression promotes EMT in prostate cancer cells via MMP2/MMP9-mediated shedding of HB-EGF, causing constitutive EGFR phosphorylation and downstream ERK signaling.","method":"LIV-1 overexpression, zymography for MMP2/MMP9 activity, EGFR phosphorylation by Western blot, ERK activation assay","journal":"PloS one","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, gain-of-function with pathway readouts but limited mechanistic dissection of HB-EGF specifically","pmids":["22110740"],"is_preprint":false},{"year":2012,"finding":"miR-1207-5p directly regulates HBEGF expression in podocytes via its 3'UTR; a miRSNP C1936T at position 2 of the seed region abolishes this regulation, as shown by luciferase reporter assay and western blot with miRNA mimics.","method":"Luciferase reporter assay with 3'UTR construct, miRNA mimic transfection, Western blot for HBEGF protein in podocytes","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct reporter and protein-level validation of miRNA-target interaction with SNP functional analysis, single lab","pmids":["22319602"],"is_preprint":false},{"year":2013,"finding":"Hypoxia increases ADAM12 levels and activity in a Notch signaling-dependent manner, leading to increased HB-EGF ectodomain shedding; released HB-EGF induces invadopodium formation in cancer cells, linking Notch and EGFR pathways in promoting invasion.","method":"Hypoxia treatment, Notch pathway manipulation, ADAM12 activity assay, HB-EGF shedding measurement, invadopodium formation assay","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and pharmacological Notch manipulation linked to ADAM12-HB-EGF-invadopodia axis, single lab","pmids":["23589494"],"is_preprint":false},{"year":2013,"finding":"Autocrine HB-EGF expression in breast cancer cells enhances invadopodium formation and function via EGFR signaling, and upregulates MMP2 and MMP9; HB-EGF inhibition rapidly decreased invadopodia, demonstrating direct EGFR-dependent mechanism.","method":"HB-EGF overexpression, EGFR inhibition, in vivo intravasation assay, MMP2/9 expression analysis, invadopodium formation assay","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain-of-function with EGFR inhibitor rescue and in vivo confirmation, single lab","pmids":["24013225"],"is_preprint":false},{"year":2014,"finding":"miR-96 promotes osteogenic differentiation by suppressing HB-EGF post-transcriptionally via binding to the 3'UTR of HB-EGF mRNA, thereby reducing EGFR and ERK1/AKT phosphorylation downstream.","method":"miR-96 overexpression in MC3T3-E1 cells, 3'UTR binding assay, Western blot for HB-EGF/pEGFR/pERK1/pAKT, osteogenic differentiation markers","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct 3'UTR binding with downstream pathway validation, single lab","pmids":["25451232"],"is_preprint":false},{"year":2014,"finding":"HB-EGF promotes intestinal epithelial cell migration and adhesion via FAK phosphorylation; HB-EGF increased p-FAK expression and induced FAK redistribution and actin reorganization, and FAK inhibitor reversed HB-EGF-induced migration and adhesion.","method":"Scrape wound healing assay, FAK inhibitor 14, immunofluorescence for p-FAK redistribution, Western blot, fibronectin adhesion assay","journal":"The Journal of surgical research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological FAK inhibition with multiple functional readouts, single lab","pmids":["24703506"],"is_preprint":false},{"year":2015,"finding":"In head and neck squamous cell carcinoma, Abl kinases negatively regulate invadopodia through suppression of an HB-EGF autocrine loop; imatinib stimulated HB-EGF shedding from HNSCC cells, and soluble HB-EGF enhanced invadopodia ECM degradation via an EGFR-Src-cortactin cascade.","method":"Imatinib treatment, HB-EGF shedding measurement, Abl/Arg siRNA, invadopodium formation and ECM degradation assays, EGFR/Src inhibitors","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA and pharmacological inhibitors establishing pathway context, single lab","pmids":["23146907"],"is_preprint":false},{"year":2015,"finding":"Recombinant ADAM12 prodomain (PA12) selectively inhibits ADAM12 but not ADAM10 or ADAM17, reducing HB-EGF shedding and cellular migration in endometriotic cells; computational modeling predicted that high-affinity ligands like HB-EGF are more difficult to target with decoy antibodies than low-affinity ligands.","method":"Recombinant ADAM12 prodomain protein, HB-EGF shedding assay, selectivity profiling against ADAM10/17, cell migration assay, computational modeling","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro biochemical inhibition with selectivity profiling plus functional validation, single lab","pmids":["26477568"],"is_preprint":false},{"year":2017,"finding":"HBEGF can initiate glioblastoma in mice with Ink4a/Arf and Pten loss; HBEGF stimulation activates both EGFR and the RTK Axl, and EGFR is required for Axl activation; silencing HBEGF in vivo caused tumor regression.","method":"In vivo mouse GBM model, EGFR/Axl conditional deletion, in vivo HBEGF silencing with survival analysis","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo genetic model with epistasis between EGFR and Axl, single lab","pmids":["28368403"],"is_preprint":false},{"year":2018,"finding":"HB-EGF gene expression in β-cells is increased by glucose in a ChREBP-dependent manner; ChREBP binding sites near the HB-EGF gene were identified by ChIP; HB-EGF knockdown in rat islets blocked glucose-induced β-cell proliferation ex vivo and in vivo; Src family kinase inhibition abrogated glucose-induced proliferation, implicating Src in HB-EGF processing.","method":"HB-EGF siRNA knockdown in isolated islets, transplanted glucose-infused rats, ChREBP ChIP, EGFR and HB-EGF inhibitors, Src kinase inhibitors, β-cell proliferation assay","journal":"Diabetes","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP for transcriptional regulation, in vitro and in vivo KD with functional rescue, multiple inhibitors, single lab with multiple orthogonal approaches","pmids":["31882563"],"is_preprint":false},{"year":2018,"finding":"BHLHE40 induces HBEGF transcription by blocking DNA binding of HDAC1 and HDAC2, as shown by ChIP and Co-IP assays; HBEGF is then secreted through exosomes to promote cell survival and migration.","method":"ChIP assay, Co-IP of BHLHE40 with HDAC1/2, exosome analysis, HBEGF knockdown, migration/survival assays, xenograft and experimental metastasis models","journal":"Breast cancer research : BCR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and Co-IP identify transcriptional mechanism, functional validation in vivo, single lab","pmids":["30285805"],"is_preprint":false},{"year":2019,"finding":"Myeloid cell-derived HB-EGF induces epithelial EGFR nuclear translocation and methylation of histone H4 to facilitate DNA damage repair in pancreatic acinar cells; conditional knockout of HB-EGF in myeloid cells (LysM-Cre) delayed recovery from pancreatitis with impaired DNA repair and increased apoptosis.","method":"Conditional myeloid-specific Hbegf KO (LysM-Cre), myeloid cell depletion (CD11b-DTR), EGFR conditional KO in pancreatic epithelium, H4 methylation immunostaining, DNA damage assays, cell proliferation/apoptosis measurements","journal":"Cellular and molecular gastroenterology and hepatology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple conditional KO models converging on HB-EGF/EGFR/H4 methylation axis, in vitro mechanistic validation","pmids":["31125624"],"is_preprint":false},{"year":2019,"finding":"s-HBEGF released from sorafenib-treated vascular endothelial cells activates EGFR on keratinocytes and promotes JNK2-mediated SIRT1 stabilization, driving hyper-keratinization; HBEGF neutralization, SIRT1 knockdown, or nicotinamide (SIRT1 inhibitor) all reduced sorafenib-induced hand-foot skin reaction in mouse models.","method":"In vivo mouse HFSR model, s-HBEGF administration, HBEGF neutralization antibody, SIRT1 siRNA knockdown, nicotinamide treatment, pJNK2 and SIRT1 Western blot, preliminary clinical study","journal":"Cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo model with genetic and pharmacological interventions defining s-HBEGF→EGFR→JNK2→SIRT1 pathway, single lab","pmids":["32296111"],"is_preprint":false},{"year":2019,"finding":"HBEGF+ inflammatory macrophages in rheumatoid arthritis promote fibroblast invasiveness in an EGF receptor-dependent manner; this cross-talk is shaped by resident fibroblasts and TNF.","method":"Single-cell RNA sequencing, ex vivo synovial tissue assay with EGFR pathway inhibition, functional invasiveness assays","journal":"Science translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — scRNA-seq identification plus functional ex vivo validation with EGFR-dependence shown, single study","pmids":["31068444"],"is_preprint":false},{"year":2020,"finding":"TMPRSS4 increases both transcriptional/translational levels of HB-EGF precursor and promotes its proteolytic cleavage by enhancing MMP9 expression via EGFR/Akt/mTOR/HIF-1α signaling; HB-EGF in turn promotes HCC proliferation and invasion via EGFR/PI3K/Akt pathway.","method":"TMPRSS4 overexpression/knockdown, MMP9 activity assay, EGFR/Akt/mTOR pathway inhibitors, CRM-197 HB-EGF inhibitor, in vivo xenograft model","journal":"Hepatology (Baltimore, Md.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain/loss-of-function with pathway inhibitors in vitro and in vivo, single lab","pmids":["31867749"],"is_preprint":false},{"year":2020,"finding":"APP interacts with pro-HB-EGF (identified in yeast two-hybrid screen and confirmed by co-immunoprecipitation); APP and EGF synergistically activate ERK signaling and promote neuritogenesis via EGFR, as shown by EGFR inhibitor PD 168393 abrogation.","method":"Yeast two-hybrid screen, co-immunoprecipitation, EGFR inhibitor, immunofluorescence neurite analysis, Western blot for pERK","journal":"Molecular neurobiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — yeast two-hybrid plus Co-IP confirms interaction; functional EGFR-dependence established by inhibitor, single lab","pmids":["33009641"],"is_preprint":false},{"year":2022,"finding":"Group 3 innate lymphoid cells (ILC3s) produce HB-EGF in response to prostaglandin E2/EP2 receptor engagement; ILC3-derived HB-EGF protects intestinal epithelium from TNF-induced cell death independently of IL-22; mice lacking ILC3-derived HB-EGF showed increased susceptibility to TNF-mediated epithelial death and experimental intestinal inflammation.","method":"Conditional HB-EGF KO in ILC3s (Hbegf f/f × ILC3-Cre), PGE2 stimulation assays, TNF-induced intestinal injury model, in vitro epithelial protection assays","journal":"Nature immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — cell-type-specific conditional KO with in vivo and in vitro mechanistic dissection, multiple orthogonal approaches","pmids":["35102343"],"is_preprint":false},{"year":2022,"finding":"Combined TNF and HBEGF treatment in human retinal organoids induces photoreceptor neurodegeneration via a previously unknown cell extrusion mechanism; pharmacological inhibitors of PIEZO1 (mechanosensor), MAPK, and actomyosin each prevented pathogenesis, and a PIEZO1 activator alone induced photoreceptor extrusion.","method":"Human retinal organoid system, combined HBEGF+TNF treatment, PIEZO1 pharmacological activation and inhibition, MAPK/actomyosin inhibitors, live imaging, transcriptome analysis","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — organoid model with pharmacological pathway dissection revealing novel mechanism, single study","pmids":["36261438"],"is_preprint":false},{"year":2023,"finding":"Hb-egfa produced by ependymal cells is required for spinal cord regeneration in zebrafish; hb-egfa mutants show defective axon crossing, tissue bridging, and swim recovery after transection; local recombinant HB-EGF delivery alters ependymal cell cycling and enhances functional regeneration; a tissue regeneration enhancer element (TREE) linked to hb-egfa drives gene expression specifically at spinal cord injuries.","method":"hb-egfa mutant zebrafish, recombinant HB-EGF delivery, epigenetic profiling for TREE identification, AAV-based enhancer-driven delivery in neonatal mouse spinal cord crush, BrdU/EdU cell cycling assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic mutant plus recombinant protein rescue plus epigenetic TREE characterization with cross-species validation in mouse","pmids":["37567873"],"is_preprint":false},{"year":2024,"finding":"Astrocyte-derived HB-EGF limits autoimmune CNS pathology; hypoxic conditions rapidly upregulate HB-EGF in astrocytes, while pro-inflammatory conditions suppress HB-EGF signaling through epigenetic modifications; intranasal HB-EGF administration attenuated disease in a mouse EAE model.","method":"Proteomic analysis of CSF, in vitro cell type studies, epigenetic modification analysis of HB-EGF locus, intranasal HB-EGF delivery in EAE model","journal":"Nature immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro mechanistic studies plus in vivo therapeutic validation, but epigenetic details are limited in the abstract, single study","pmids":["38409259"],"is_preprint":false}],"current_model":"HBEGF encodes a dual-form growth factor (membrane-anchored proHB-EGF and shed soluble HB-EGF) that binds and activates EGFR/ErbB4; its ectodomain is shed by ADAM10, ADAM12, or ADAM17 (TACE) in response to diverse stimuli including GPCR agonists, mechanical stress, wounding, and hypoxia, generating a paracrine/autocrine signal that activates ERK, PI3K/Akt, and downstream transcriptional programs to drive cardiac valve morphogenesis, epithelial wound closure, β-cell proliferation, intestinal repair, neurogenesis, and cancer invasion, while membrane-retained proHB-EGF engages in juxtacrine signaling by complexing with CD9 and integrin α3β1 at cell-cell contacts."},"narrative":{"mechanistic_narrative":"HBEGF encodes a dual-form EGFR ligand that exists as a membrane-anchored precursor (proHB-EGF) and a proteolytically shed soluble form, and these two forms drive opposing cellular programs across morphogenesis, tissue repair, and cancer [PMID:15289334, PMID:14996914]. Membrane-anchored proHB-EGF localizes to cell-cell contacts in a complex with the tetraspanin CD9 and integrin α3β1, where CD9 upregulates its juxtacrine activity, and its heparin-binding domain anchors it at contact sites through trans interaction with heparan sulfate proteoglycans; disrupting this interaction promotes proteolytic release and switches behavior from juxtacrine growth inhibition to autocrine proliferation [PMID:7790364, PMID:9514697, PMID:20530570]. Ectodomain shedding is executed by metalloproteases—ADAM12 in cardiomyocytes, ADAM17/TACE downstream of GPCRs, and MMP-7/MMP2/MMP9 in epithelial and cancer contexts—and is triggered by GPCR agonists (estrogen via GPR30, angiotensin II, 5-HT2A), mechanical stretch, wounding, and hypoxia/Notch signaling, with ERK1/2 feeding back to phosphorylate and activate ADAM17 [PMID:11043579, PMID:11786904, PMID:16737974, PMID:18658095, PMID:19237431, PMID:23589494]. Soluble HB-EGF transactivates EGFR to engage ERK and PI3K/Akt cascades, driving cardiac valve morphogenesis through suppression of BMP/Smad signaling, eyelid and corneal epithelial closure, intestinal restitution, glucose-stimulated β-cell proliferation via ChREBP-driven transcription, and angiogenesis through a PI3K/eNOS axis [PMID:12773386, PMID:14985295, PMID:16141218, PMID:16083716, PMID:18092233, PMID:18925469, PMID:31882563]. In disease, soluble HB-EGF promotes tumorigenesis, EMT via Snail-2, and EGFR-dependent invadopodium formation, can initiate glioblastoma through coordinated EGFR and Axl activation, and mediates inflammatory tissue protection and regeneration including ILC3- and astrocyte-derived epithelial/CNS protection and zebrafish spinal cord regeneration [PMID:15289334, PMID:19244405, PMID:24013225, PMID:28368403, PMID:35102343, PMID:37567873, PMID:38409259]. HBEGF expression is controlled transcriptionally (ChREBP, BHLHE40) and post-transcriptionally (miR-1207-5p, miR-96 acting on its 3'UTR) [PMID:22319602, PMID:25451232, PMID:31882563, PMID:30285805].","teleology":[{"year":1995,"claim":"Established that the membrane-anchored precursor is not merely a latent ligand reservoir but a structured signaling unit at cell-cell contacts, defining its juxtacrine context.","evidence":"Co-IP, chemical cross-linking, and colocalization of proHB-EGF with CD9 and integrin α3β1 in Vero/A431/MG63 cells","pmids":["7790364"],"confidence":"High","gaps":["Functional consequence of the CD9/integrin complex for signaling not resolved here","Stoichiometry and structure of the complex undefined"]},{"year":1998,"claim":"Mapped which tetraspanin partner and which protein domains confer juxtacrine mitogenic enhancement, distinguishing CD9 from related tetraspanins.","evidence":"Chimeric CD9/CD81 cotransfection in LM cells with diphtheria toxin receptor functional readout","pmids":["9514697"],"confidence":"Medium","gaps":["Single lab","Mechanism by which the CD9 C-terminal region enhances activity unknown"]},{"year":2002,"claim":"Defined HB-EGF shedding as the mechanistic link in GPCR-to-EGFR transactivation across multiple receptors, converting diverse extracellular cues into EGFR signaling.","evidence":"Estrogen/GPR30, angiotensin II, and TGF-β systems using HB-EGF-AP shedding reporters, CRM-197, neutralizing antibodies, and EGFR/MMP/PKC inhibitors","pmids":["11043579","11737589","12164862"],"confidence":"High","gaps":["Specific protease assignments varied by stimulus","In vivo relevance of individual GPCR-driven shedding events not fully established"]},{"year":2002,"claim":"Identified ADAM12 as the physiological sheddase coupling GPCR agonists to EGFR transactivation in cardiomyocyte hypertrophy, providing the first defined protease-ligand-receptor cardiac axis.","evidence":"Dominant-negative ADAM12, direct KB-R7785 binding, in vivo cardiac hypertrophy model","pmids":["11786904"],"confidence":"High","gaps":["Whether ADAM12 is the sole sheddase in heart not excluded","Direct demonstration of HB-EGF as the only relevant ADAM12 substrate lacking"]},{"year":2003,"claim":"Demonstrated a developmental requirement: soluble HB-EGF activates EGFR to suppress BMP/Smad signaling during cardiac valvulogenesis, showing the genetic non-redundancy of the ligand.","evidence":"HB-EGF−/−, EGFR−/−, and TACE−/− knockout mice with convergent valve phenotype and pSmad1/5/8 staining","pmids":["12773386"],"confidence":"High","gaps":["Direct biochemical link between EGFR activation and Smad suppression not delineated","Cell-type source of HB-EGF in valve not pinpointed"]},{"year":2004,"claim":"Distinguished the opposing functions of the two HB-EGF forms—soluble form is tumorigenic and pro-migratory while membrane form promotes adhesion and inhibits migration—establishing form-specific biology.","evidence":"Inducible and stable expression of soluble vs noncleavable proHB-EGF in bladder cancer and MDCK cells; xenograft, colony, migration, and adhesion assays","pmids":["15289334","14996914"],"confidence":"High","gaps":["Molecular basis of membrane-form growth inhibition not fully resolved","Single-cell-line dependence of some readouts"]},{"year":2005,"claim":"Showed HB-EGF drives epithelial wound closure and restitution through EGFR-ERK and PI3K/Akt, defining the effector cascades for tissue repair.","evidence":"Corneal/eyelid wound models with HB-EGF null and secretion-deficient mice, EGFR hypomorphs, and intestinal I/R with pathway inhibitors","pmids":["14985295","16141218","16083716"],"confidence":"High","gaps":["Relative contribution of ERK vs PI3K branches tissue-dependent","Identity of sheddase in each wound context not uniformly defined"]},{"year":2006,"claim":"Extended the sheddase repertoire to TACE/ADAM17 and MMP-7 for specific GPCR and bile-acid stimuli, showing protease usage is stimulus- and tissue-specific.","evidence":"TACE siRNA and Co-IP with HB-EGF in mesangial cells; MMP-7 siRNA/neutralizing antibody and colocalization in colon cancer cells","pmids":["16737974","17222808"],"confidence":"Medium","gaps":["Single lab per system","Direct cleavage by these proteases not reconstituted biochemically"]},{"year":2008,"claim":"Revealed a feedback loop in which ERK1/2 phosphorylates and activates ADAM17, amplifying HB-EGF shedding and EGFR transactivation.","evidence":"MEK inhibitors, HB-EGF-AP shedding assay, ERK-ADAM17 Co-IP and serine phosphorylation in corneal epithelial cells","pmids":["18658095"],"confidence":"High","gaps":["Phosphosite mapping incomplete","Generality beyond corneal epithelium untested"]},{"year":2009,"claim":"Defined how proHB-EGF is retained at contacts and how its release is gated, linking heparan sulfate binding to the juxtacrine-to-autocrine switch.","evidence":"Dual-domain tracking, heparin/HSPG disruption, and proliferation/wound assays; integrin/FAK masking of HB-EGF growth activity in 2D vs 3D","pmids":["20530570","19887590"],"confidence":"Medium","gaps":["Single lab","In vivo relevance of HSPG-mediated retention not demonstrated"]},{"year":2009,"claim":"Connected HB-EGF to EMT and mechanotransduction-driven differentiation, broadening its program beyond proliferation.","evidence":"sHB-EGF overexpression with Snail-2 knockdown in renal cells; AP-tagged HB-EGF shedding under cyclic stretch in fetal type II cells","pmids":["19244405","19237431"],"confidence":"Medium","gaps":["Single lab","Receptor-level coupling to Snail-2 induction not mapped"]},{"year":2013,"claim":"Placed HB-EGF at the center of cancer invasion machinery, linking hypoxia/Notch-induced ADAM12 shedding and autocrine EGFR signaling to invadopodium formation.","evidence":"Hypoxia/Notch manipulation, ADAM12 activity, and invadopodium assays in cancer cells; autocrine HB-EGF overexpression and EGFR inhibition with in vivo intravasation","pmids":["23589494","24013225"],"confidence":"Medium","gaps":["Single lab per study","Quantitative contribution to metastasis in vivo limited"]},{"year":2015,"claim":"Profiled selective ADAM12 inhibition and Abl-mediated negative regulation of the HB-EGF autocrine loop, identifying therapeutic targeting and regulatory nodes.","evidence":"Recombinant ADAM12 prodomain selectivity profiling and migration assay; imatinib-induced HB-EGF shedding and EGFR-Src-cortactin cascade in HNSCC","pmids":["26477568","23146907"],"confidence":"Medium","gaps":["High-affinity HB-EGF predicted hard to target by decoys","Single lab per study"]},{"year":2017,"claim":"Showed HB-EGF can act as a tumor initiator through coordinated activation of EGFR and the RTK Axl, with EGFR upstream of Axl.","evidence":"In vivo GBM model with Ink4a/Arf and Pten loss, conditional EGFR/Axl deletion, in vivo HBEGF silencing","pmids":["28368403"],"confidence":"Medium","gaps":["Mechanism of EGFR-dependent Axl activation undefined","Single model system"]},{"year":2018,"claim":"Established the regulatory inputs controlling HBEGF gene expression, both transcriptional (ChREBP, BHLHE40 via HDAC blockade) and post-transcriptional (miR-1207-5p, miR-96), linking nutrient and oncogenic signals to ligand abundance.","evidence":"ChREBP and BHLHE40 ChIP/Co-IP with islet and breast cancer functional assays; luciferase 3'UTR reporters and miRNA mimics in podocytes and osteoblasts","pmids":["31882563","30285805","22319602","25451232"],"confidence":"High","gaps":["Interplay among these regulators untested","Tissue-specificity of each control mechanism partially defined"]},{"year":2019,"claim":"Identified immune- and stroma-derived HB-EGF as a driver of tissue repair and pathology, including myeloid HB-EGF promoting DNA repair via EGFR nuclear translocation and macrophage HB-EGF driving fibroblast invasiveness.","evidence":"Conditional myeloid Hbegf KO with H4 methylation/DNA repair assays in pancreatitis; scRNA-seq and ex vivo EGFR-inhibition assays in rheumatoid arthritis synovium","pmids":["31125624","31068444"],"confidence":"High","gaps":["Mechanism of EGFR nuclear translocation incompletely defined","Direct sheddase in immune contexts not specified"]},{"year":2020,"claim":"Expanded the receptor and partner network, showing APP physically binds proHB-EGF and synergizes with EGF in neuritogenesis, and that TMPRSS4 amplifies both HB-EGF precursor levels and its cleavage in HCC.","evidence":"Yeast two-hybrid and Co-IP for APP-proHB-EGF with EGFR-dependent neurite assays; TMPRSS4 gain/loss with MMP9 and EGFR/PI3K/Akt readouts in HCC","pmids":["33009641","31867749"],"confidence":"Medium","gaps":["Functional significance of APP interaction in vivo unknown","Single lab per study"]},{"year":2022,"claim":"Defined protective and pathogenic context-dependent roles of HB-EGF in mucosal and neural tissue, from ILC3-derived epithelial protection to PIEZO1-coupled photoreceptor extrusion.","evidence":"ILC3-specific Hbegf conditional KO with TNF-injury models; human retinal organoid HBEGF+TNF treatment with PIEZO1/MAPK/actomyosin inhibitors","pmids":["35102343","36261438"],"confidence":"High","gaps":["Receptor coupling for ILC3-derived protection not fully mapped","Organoid extrusion mechanism single study"]},{"year":2024,"claim":"Demonstrated regenerative and immunoprotective HB-EGF functions in the CNS, with ependymal HB-EGF required for spinal cord regeneration and astrocyte-derived HB-EGF limiting autoimmune pathology.","evidence":"hb-egfa zebrafish mutants with recombinant rescue and TREE characterization in mouse; CSF proteomics and intranasal HB-EGF in EAE","pmids":["37567873","38409259"],"confidence":"High","gaps":["Downstream effector pathways in regeneration not detailed","Epigenetic control of astrocyte HB-EGF only partially characterized"]},{"year":null,"claim":"How the choice between juxtacrine proHB-EGF signaling and autocrine/paracrine soluble HB-EGF signaling is integrated at the level of a single cell—and which sheddase dominates in each physiological context in vivo—remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model of context-specific sheddase selection in vivo","Structural basis of the proHB-EGF/CD9/integrin complex undefined","Quantitative balance between membrane-form inhibition and soluble-form activation not established"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[2,7,8,9,13]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[19]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[2,7,13,33]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1,15,19]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[9,8,34]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,3,13,14,17]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[7,12,42]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[9,27,32,38]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[37,40,43]}],"complexes":["proHB-EGF/CD9/integrin α3β1 complex"],"partners":["EGFR","CD9","ITGA3","ADAM12","ADAM17","MMP7","APP","AXL"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q99075","full_name":"Proheparin-binding EGF-like growth factor","aliases":[],"length_aa":208,"mass_kda":23.1,"function":"Growth factor that mediates its effects via EGFR, ERBB2 and ERBB4. Required for normal cardiac valve formation and normal heart function. Promotes smooth muscle cell proliferation. May be involved in macrophage-mediated cellular proliferation. It is mitogenic for fibroblasts, but not endothelial cells. It is able to bind EGF receptor/EGFR with higher affinity than EGF itself and is a far more potent mitogen for smooth muscle cells than EGF. Also acts as a diphtheria toxin receptor","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q99075/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/HBEGF","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/HBEGF","total_profiled":1310},"omim":[{"mim_id":"620290","title":"TRANSMEMBRANE PROTEIN 219; TMEM219","url":"https://www.omim.org/entry/620290"},{"mim_id":"614404","title":"RHOMBOID 5 HOMOLOG 2; RHBDF2","url":"https://www.omim.org/entry/614404"},{"mim_id":"611288","title":"CORNICHON FAMILY AMPA RECEPTOR AUXILIARY PROTEIN 2; CNIH2","url":"https://www.omim.org/entry/611288"},{"mim_id":"610207","title":"SOLUTE CARRIER FAMILY 4 (ANION EXCHANGER), MEMBER 9; SLC4A9","url":"https://www.omim.org/entry/610207"},{"mim_id":"610134","title":"ST6 ALPHA-N-ACETYL-NEURAMINYL-2,3-BETA-GALACTOSYL-1,3-N-ACETYLGALACTOSAMINIDE ALPHA-2,6-SIALYLTRANSFERASE 5; ST6GALNAC5","url":"https://www.omim.org/entry/610134"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Golgi apparatus","reliability":"Approved"},{"location":"Vesicles","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"urinary bladder","ntpm":119.6}],"url":"https://www.proteinatlas.org/search/HBEGF"},"hgnc":{"alias_symbol":[],"prev_symbol":["HEGFL","DTS","DTR"]},"alphafold":{"accession":"Q99075","domains":[{"cath_id":"2.10.25.10","chopping":"108-147","consensus_level":"high","plddt":91.525,"start":108,"end":147}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q99075","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q99075-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q99075-F1-predicted_aligned_error_v6.png","plddt_mean":66.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=HBEGF","jax_strain_url":"https://www.jax.org/strain/search?query=HBEGF"},"sequence":{"accession":"Q99075","fasta_url":"https://rest.uniprot.org/uniprotkb/Q99075.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q99075/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q99075"}},"corpus_meta":[{"pmid":"11043579","id":"PMC_11043579","title":"Estrogen-induced activation of Erk-1 and Erk-2 requires the G protein-coupled receptor homolog, GPR30, and occurs via trans-activation of the epidermal growth factor receptor through release of HB-EGF.","date":"2000","source":"Molecular endocrinology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/11043579","citation_count":1166,"is_preprint":false},{"pmid":"11786904","id":"PMC_11786904","title":"Cardiac hypertrophy is inhibited by antagonism of ADAM12 processing of HB-EGF: metalloproteinase inhibitors as a new therapy.","date":"2002","source":"Nature medicine","url":"https://pubmed.ncbi.nlm.nih.gov/11786904","citation_count":606,"is_preprint":false},{"pmid":"12773386","id":"PMC_12773386","title":"Defective valvulogenesis in HB-EGF and TACE-null mice is associated with aberrant BMP signaling.","date":"2003","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/12773386","citation_count":321,"is_preprint":false},{"pmid":"12882410","id":"PMC_12882410","title":"Neurogenesis and aging: FGF-2 and HB-EGF restore neurogenesis in hippocampus and subventricular zone of aged mice.","date":"2003","source":"Aging cell","url":"https://pubmed.ncbi.nlm.nih.gov/12882410","citation_count":314,"is_preprint":false},{"pmid":"7790364","id":"PMC_7790364","title":"Membrane-anchored heparin-binding EGF-like growth factor (HB-EGF) and diphtheria toxin receptor-associated protein (DRAP27)/CD9 form a complex with integrin alpha 3 beta 1 at cell-cell contact sites.","date":"1995","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/7790364","citation_count":234,"is_preprint":false},{"pmid":"31068444","id":"PMC_31068444","title":"HBEGF+ macrophages in rheumatoid arthritis induce fibroblast invasiveness.","date":"2019","source":"Science translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31068444","citation_count":205,"is_preprint":false},{"pmid":"15289334","id":"PMC_15289334","title":"HB-EGF is a potent inducer of tumor growth and angiogenesis.","date":"2004","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/15289334","citation_count":177,"is_preprint":false},{"pmid":"16054021","id":"PMC_16054021","title":"ADAM-mediated ectodomain shedding of HB-EGF in receptor cross-talk.","date":"2004","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/16054021","citation_count":136,"is_preprint":false},{"pmid":"9514697","id":"PMC_9514697","title":"Functional analysis of four tetraspans, CD9, CD53, CD81, and CD82, suggests a common role in costimulation, cell adhesion, and migration: only CD9 upregulates HB-EGF activity.","date":"1997","source":"Cellular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/9514697","citation_count":128,"is_preprint":false},{"pmid":"18023415","id":"PMC_18023415","title":"Validation of HB-EGF and amphiregulin as targets for human cancer therapy.","date":"2007","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/18023415","citation_count":126,"is_preprint":false},{"pmid":"18092233","id":"PMC_18092233","title":"HB-EGF promotes angiogenesis in endothelial cells via PI3-kinase and MAPK signaling pathways.","date":"2007","source":"Growth factors (Chur, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/18092233","citation_count":124,"is_preprint":false},{"pmid":"14985295","id":"PMC_14985295","title":"Wound-induced HB-EGF ectodomain shedding and EGFR activation in corneal epithelial cells.","date":"2004","source":"Investigative ophthalmology & visual science","url":"https://pubmed.ncbi.nlm.nih.gov/14985295","citation_count":124,"is_preprint":false},{"pmid":"23589494","id":"PMC_23589494","title":"Notch increases the shedding of HB-EGF by ADAM12 to potentiate invadopodia formation in hypoxia.","date":"2013","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/23589494","citation_count":120,"is_preprint":false},{"pmid":"16508205","id":"PMC_16508205","title":"ErbB and HB-EGF signaling in heart development and function.","date":"2006","source":"Cell structure and function","url":"https://pubmed.ncbi.nlm.nih.gov/16508205","citation_count":113,"is_preprint":false},{"pmid":"12099696","id":"PMC_12099696","title":"Helicobacter pylori-stimulated EGF receptor transactivation requires metalloprotease cleavage of HB-EGF.","date":"2002","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/12099696","citation_count":106,"is_preprint":false},{"pmid":"14697362","id":"PMC_14697362","title":"HB-EGF directs stromal cell polyploidy and decidualization via cyclin D3 during implantation.","date":"2004","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/14697362","citation_count":99,"is_preprint":false},{"pmid":"16737974","id":"PMC_16737974","title":"5-HT2A receptor induces ERK phosphorylation and proliferation through ADAM-17 tumor necrosis factor-alpha-converting enzyme (TACE) activation and heparin-bound epidermal growth factor-like growth factor (HB-EGF) shedding in mesangial cells.","date":"2006","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16737974","citation_count":95,"is_preprint":false},{"pmid":"16083716","id":"PMC_16083716","title":"HB-EGF enhances restitution after intestinal ischemia/reperfusion via PI3K/Akt and MEK/ERK1/2 activation.","date":"2005","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/16083716","citation_count":95,"is_preprint":false},{"pmid":"10510187","id":"PMC_10510187","title":"Multiple trophic actions of heparin-binding epidermal growth factor (HB-EGF) in the central nervous system.","date":"1999","source":"The European journal of neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/10510187","citation_count":93,"is_preprint":false},{"pmid":"22110740","id":"PMC_22110740","title":"LIV-1 promotes prostate cancer epithelial-to-mesenchymal transition and metastasis through HB-EGF shedding and EGFR-mediated ERK signaling.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22110740","citation_count":92,"is_preprint":false},{"pmid":"12112577","id":"PMC_12112577","title":"Regulated expression of heparin-binding EGF-like growth factor (HB-EGF) in the human endometrium: a potential paracrine role during implantation.","date":"2002","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/12112577","citation_count":90,"is_preprint":false},{"pmid":"16141218","id":"PMC_16141218","title":"HB-EGF promotes epithelial cell migration in eyelid development.","date":"2005","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/16141218","citation_count":85,"is_preprint":false},{"pmid":"24013225","id":"PMC_24013225","title":"Autocrine HBEGF expression promotes breast cancer intravasation, metastasis and macrophage-independent invasion in vivo.","date":"2013","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/24013225","citation_count":81,"is_preprint":false},{"pmid":"12164862","id":"PMC_12164862","title":"Involvement of HB-EGF and EGF receptor transactivation in TGF-beta-mediated fibronectin expression in mesangial cells.","date":"2002","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/12164862","citation_count":80,"is_preprint":false},{"pmid":"19064678","id":"PMC_19064678","title":"HBEGF, SRA1, and IK: Three cosegregating genes as determinants of cardiomyopathy.","date":"2008","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/19064678","citation_count":79,"is_preprint":false},{"pmid":"35102343","id":"PMC_35102343","title":"Group 3 innate lymphoid cells produce the growth factor HB-EGF to protect the intestine from TNF-mediated inflammation.","date":"2022","source":"Nature immunology","url":"https://pubmed.ncbi.nlm.nih.gov/35102343","citation_count":71,"is_preprint":false},{"pmid":"17222808","id":"PMC_17222808","title":"Matrix metalloproteinase-7-catalyzed release of HB-EGF mediates deoxycholyltaurine-induced proliferation of a human colon cancer cell line.","date":"2006","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/17222808","citation_count":66,"is_preprint":false},{"pmid":"11737589","id":"PMC_11737589","title":"Angiotensin II signaling and HB-EGF shedding via metalloproteinase in glomerular mesangial cells.","date":"2001","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/11737589","citation_count":61,"is_preprint":false},{"pmid":"35342363","id":"PMC_35342363","title":"HB-EGF induces mitochondrial dysfunction via estrogen hypersecretion in granulosa cells dependent on cAMP-PKA-JNK/ERK-Ca2+-FOXO1 pathway.","date":"2022","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35342363","citation_count":60,"is_preprint":false},{"pmid":"18925469","id":"PMC_18925469","title":"HB-EGF stimulates eNOS expression and nitric oxide production and promotes eNOS dependent angiogenesis.","date":"2008","source":"Growth factors (Chur, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/18925469","citation_count":60,"is_preprint":false},{"pmid":"14576062","id":"PMC_14576062","title":"An inhibitor of the EGF receptor family blocks myeloma cell growth factor activity of HB-EGF and potentiates dexamethasone or anti-IL-6 antibody-induced apoptosis.","date":"2003","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/14576062","citation_count":60,"is_preprint":false},{"pmid":"10769639","id":"PMC_10769639","title":"Increased expression of heparin binding EGF (HB-EGF), amphiregulin, TGF alpha and epiregulin in androgen-independent prostate cancer cell lines.","date":"2000","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/10769639","citation_count":60,"is_preprint":false},{"pmid":"16084404","id":"PMC_16084404","title":"Heparin-binding EGF-like growth factor (HB-EGF) and necrotizing enterocolitis.","date":"2005","source":"Seminars in pediatric surgery","url":"https://pubmed.ncbi.nlm.nih.gov/16084404","citation_count":59,"is_preprint":false},{"pmid":"18658095","id":"PMC_18658095","title":"ERK1/2 mediate wounding- and G-protein-coupled receptor ligands-induced EGFR activation via regulating ADAM17 and HB-EGF shedding.","date":"2008","source":"Investigative ophthalmology & visual science","url":"https://pubmed.ncbi.nlm.nih.gov/18658095","citation_count":58,"is_preprint":false},{"pmid":"19565643","id":"PMC_19565643","title":"Diverse functions of HBEGF during pregnancy.","date":"2009","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/19565643","citation_count":55,"is_preprint":false},{"pmid":"9473209","id":"PMC_9473209","title":"Structure and function of heparin-binding EGF-like growth factor (HB-EGF).","date":"1998","source":"Frontiers in bioscience : a journal and virtual library","url":"https://pubmed.ncbi.nlm.nih.gov/9473209","citation_count":54,"is_preprint":false},{"pmid":"17395697","id":"PMC_17395697","title":"Epidermal growth factor (EGF) receptor ligands in the chicken ovary: I. Evidence for heparin-binding EGF-like growth factor (HB-EGF) as a potential oocyte-derived signal to control granulosa cell proliferation and HB-EGF and kit ligand expression.","date":"2007","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/17395697","citation_count":54,"is_preprint":false},{"pmid":"30285805","id":"PMC_30285805","title":"BHLHE40 confers a pro-survival and pro-metastatic phenotype to breast cancer cells by modulating HBEGF secretion.","date":"2018","source":"Breast cancer research : BCR","url":"https://pubmed.ncbi.nlm.nih.gov/30285805","citation_count":53,"is_preprint":false},{"pmid":"31867749","id":"PMC_31867749","title":"TMPRSS4 Drives Angiogenesis in Hepatocellular Carcinoma by Promoting HB-EGF Expression and Proteolytic Cleavage.","date":"2020","source":"Hepatology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/31867749","citation_count":52,"is_preprint":false},{"pmid":"17490650","id":"PMC_17490650","title":"Injury and nucleotides induce phosphorylation of epidermal growth factor receptor: MMP and HB-EGF dependent pathway.","date":"2007","source":"Experimental eye research","url":"https://pubmed.ncbi.nlm.nih.gov/17490650","citation_count":47,"is_preprint":false},{"pmid":"23349855","id":"PMC_23349855","title":"Control of human endometrial stromal cell motility by PDGF-BB, HB-EGF and trophoblast-secreted factors.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23349855","citation_count":47,"is_preprint":false},{"pmid":"14996914","id":"PMC_14996914","title":"Membrane-associated HB-EGF modulates HGF-induced cellular responses in MDCK cells.","date":"2004","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/14996914","citation_count":45,"is_preprint":false},{"pmid":"22319602","id":"PMC_22319602","title":"A miR-1207-5p binding site polymorphism abolishes regulation of HBEGF and is associated with disease severity in CFHR5 nephropathy.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22319602","citation_count":44,"is_preprint":false},{"pmid":"36261438","id":"PMC_36261438","title":"HBEGF-TNF induce a complex outer retinal pathology with photoreceptor cell extrusion in human organoids.","date":"2022","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/36261438","citation_count":43,"is_preprint":false},{"pmid":"11889210","id":"PMC_11889210","title":"In vivo and in vitro evidence suggest that HB-EGF regulates endometrial expression of human decay-accelerating factor.","date":"2002","source":"The Journal of clinical endocrinology and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/11889210","citation_count":43,"is_preprint":false},{"pmid":"24188029","id":"PMC_24188029","title":"HB-EGF affects astrocyte morphology, proliferation, differentiation, and the expression of intermediate filament proteins.","date":"2013","source":"Journal of neurochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24188029","citation_count":42,"is_preprint":false},{"pmid":"15856048","id":"PMC_15856048","title":"Local overexpression of HB-EGF exacerbates remodeling following myocardial infarction by activating noncardiomyocytes.","date":"2005","source":"Laboratory investigation; a journal of technical methods and pathology","url":"https://pubmed.ncbi.nlm.nih.gov/15856048","citation_count":42,"is_preprint":false},{"pmid":"12676768","id":"PMC_12676768","title":"Vanadium-induced HB-EGF expression in human lung fibroblasts is oxidant dependent and requires MAP kinases.","date":"2003","source":"American journal of physiology. Lung cellular and molecular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/12676768","citation_count":42,"is_preprint":false},{"pmid":"36068607","id":"PMC_36068607","title":"Identification of HBEGF+ fibroblasts in the remission of rheumatoid arthritis by integrating single-cell RNA sequencing datasets and bulk RNA sequencing datasets.","date":"2022","source":"Arthritis research & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/36068607","citation_count":41,"is_preprint":false},{"pmid":"25451232","id":"PMC_25451232","title":"miR-96 promotes osteogenic differentiation by suppressing HBEGF-EGFR signaling in osteoblastic cells.","date":"2014","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/25451232","citation_count":41,"is_preprint":false},{"pmid":"19609315","id":"PMC_19609315","title":"Metalloproteinase-mediated, context-dependent function of amphiregulin and HB-EGF in human keratinocytes and skin.","date":"2010","source":"The Journal of investigative dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/19609315","citation_count":38,"is_preprint":false},{"pmid":"25575682","id":"PMC_25575682","title":"Cypermethrin Stimulates GSK3β-Dependent Aβ and p-tau Proteins and Cognitive Loss in Young Rats: Reduced HB-EGF Signaling and Downstream Neuroinflammation as Critical Regulators.","date":"2015","source":"Molecular neurobiology","url":"https://pubmed.ncbi.nlm.nih.gov/25575682","citation_count":38,"is_preprint":false},{"pmid":"11168942","id":"PMC_11168942","title":"HB-EGF is produced in the peritoneal cavity and enhances mesothelial cell adhesion and migration.","date":"2001","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/11168942","citation_count":38,"is_preprint":false},{"pmid":"37567873","id":"PMC_37567873","title":"Spinal cord repair is modulated by the neurogenic factor Hb-egf under direction of a regeneration-associated enhancer.","date":"2023","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/37567873","citation_count":37,"is_preprint":false},{"pmid":"32322668","id":"PMC_32322668","title":"SOX2 Promotes Brain Metastasis of Breast Cancer by Upregulating the Expression of FSCN1 and HBEGF.","date":"2020","source":"Molecular therapy oncolytics","url":"https://pubmed.ncbi.nlm.nih.gov/32322668","citation_count":37,"is_preprint":false},{"pmid":"23146907","id":"PMC_23146907","title":"Ableson kinases negatively regulate invadopodia function and invasion in head and neck squamous cell carcinoma by inhibiting an HB-EGF autocrine loop.","date":"2012","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/23146907","citation_count":37,"is_preprint":false},{"pmid":"31885807","id":"PMC_31885807","title":"HB-EGF Ameliorates Oxidative Stress-Mediated Uterine Decidualization Damage.","date":"2019","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/31885807","citation_count":35,"is_preprint":false},{"pmid":"24346967","id":"PMC_24346967","title":"Synergistic effect of lung tumor-associated dendritic cell-derived HB-EGF and CXCL5 on cancer progression.","date":"2014","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/24346967","citation_count":35,"is_preprint":false},{"pmid":"25918681","id":"PMC_25918681","title":"Pretreatment levels of the serum biomarkers CEA, CYFRA 21-1, SCC and the soluble EGFR and its ligands EGF, TGF-alpha, HB-EGF in the prediction of outcome in erlotinib treated non-small-cell lung cancer patients.","date":"2015","source":"SpringerPlus","url":"https://pubmed.ncbi.nlm.nih.gov/25918681","citation_count":35,"is_preprint":false},{"pmid":"32627036","id":"PMC_32627036","title":"M2 macrophages reduce the radiosensitivity of head and neck cancer by releasing HB‑EGF.","date":"2020","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/32627036","citation_count":34,"is_preprint":false},{"pmid":"19887590","id":"PMC_19887590","title":"Integrin signal masks growth-promotion activity of HB-EGF in monolayer cell cultures.","date":"2009","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/19887590","citation_count":34,"is_preprint":false},{"pmid":"18704599","id":"PMC_18704599","title":"High expression of heparanase is significantly associated with dedifferentiation and lymph node metastasis in patients with pancreatic ductal adenocarcinomas and correlated to PDGFA and via HIF1a to HB-EGF and bFGF.","date":"2008","source":"Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract","url":"https://pubmed.ncbi.nlm.nih.gov/18704599","citation_count":33,"is_preprint":false},{"pmid":"25201063","id":"PMC_25201063","title":"MiR-212 exerts suppressive effect on SKOV3 ovarian cancer cells through targeting HBEGF.","date":"2014","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/25201063","citation_count":32,"is_preprint":false},{"pmid":"32296111","id":"PMC_32296111","title":"s-HBEGF/SIRT1 circuit-dictated crosstalk between vascular endothelial cells and keratinocytes mediates sorafenib-induced hand-foot skin reaction that can be reversed by nicotinamide.","date":"2020","source":"Cell research","url":"https://pubmed.ncbi.nlm.nih.gov/32296111","citation_count":31,"is_preprint":false},{"pmid":"19237431","id":"PMC_19237431","title":"Mechanical stretch promotes fetal type II epithelial cell differentiation via shedding of HB-EGF and TGF-alpha.","date":"2009","source":"The Journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/19237431","citation_count":31,"is_preprint":false},{"pmid":"8965173","id":"PMC_8965173","title":"Measurement of regional cerebral plasma pool and hematocrit with copper-62-labeled HSA-DTS.","date":"1996","source":"Journal of nuclear medicine : official publication, Society of Nuclear Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/8965173","citation_count":31,"is_preprint":false},{"pmid":"38409259","id":"PMC_38409259","title":"The astrocyte-produced growth factor HB-EGF limits autoimmune CNS pathology.","date":"2024","source":"Nature immunology","url":"https://pubmed.ncbi.nlm.nih.gov/38409259","citation_count":30,"is_preprint":false},{"pmid":"20530570","id":"PMC_20530570","title":"The heparin-binding domain of HB-EGF mediates localization to sites of cell-cell contact and prevents HB-EGF proteolytic release.","date":"2010","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/20530570","citation_count":30,"is_preprint":false},{"pmid":"31125624","id":"PMC_31125624","title":"Myeloid Cell-Derived HB-EGF Drives Tissue Recovery After Pancreatitis.","date":"2019","source":"Cellular and molecular gastroenterology and hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/31125624","citation_count":29,"is_preprint":false},{"pmid":"30550637","id":"PMC_30550637","title":"Roles for HB-EGF in Mesenchymal Stromal Cell Proliferation and Differentiation During Skeletal Growth.","date":"2018","source":"Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research","url":"https://pubmed.ncbi.nlm.nih.gov/30550637","citation_count":29,"is_preprint":false},{"pmid":"30451982","id":"PMC_30451982","title":"Heparin-binding epidermal growth factor (HB-EGF) drives EMT in patients with COPD: implications for disease pathogenesis and novel therapies.","date":"2018","source":"Laboratory investigation; a journal of technical methods and pathology","url":"https://pubmed.ncbi.nlm.nih.gov/30451982","citation_count":29,"is_preprint":false},{"pmid":"31882563","id":"PMC_31882563","title":"HB-EGF Signaling Is Required for Glucose-Induced Pancreatic β-Cell Proliferation in Rats.","date":"2019","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/31882563","citation_count":28,"is_preprint":false},{"pmid":"36352257","id":"PMC_36352257","title":"Particulate matter promotes cancer metastasis through increased HBEGF expression in macrophages.","date":"2022","source":"Experimental & molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36352257","citation_count":27,"is_preprint":false},{"pmid":"29379775","id":"PMC_29379775","title":"Helicobacter pylori-Induced HB-EGF Upregulates Gastrin Expression via the EGF Receptor, C-Raf, Mek1, and Erk2 in the MAPK Pathway.","date":"2018","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/29379775","citation_count":27,"is_preprint":false},{"pmid":"31648182","id":"PMC_31648182","title":"Estrogen deficiency induces memory loss via altered hippocampal HB-EGF and autophagy.","date":"2020","source":"The Journal of endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/31648182","citation_count":27,"is_preprint":false},{"pmid":"17050278","id":"PMC_17050278","title":"Expression of HB-EGF by retinal pigment epithelial cells in vitreoretinal proliferative disease.","date":"2006","source":"Current eye research","url":"https://pubmed.ncbi.nlm.nih.gov/17050278","citation_count":27,"is_preprint":false},{"pmid":"12746188","id":"PMC_12746188","title":"Upregulation of endogenous heparin-binding EGF-like growth factor (HB-EGF) expression after intestinal ischemia/reperfusion injury.","date":"2003","source":"Journal of investigative surgery : the official journal of the Academy of Surgical Research","url":"https://pubmed.ncbi.nlm.nih.gov/12746188","citation_count":27,"is_preprint":false},{"pmid":"16622802","id":"PMC_16622802","title":"Heparin-binding epidermal growth factor (HB-EGF) may improve embryonic development and implantation by increasing vitronectin receptor (integrin alphanubeta3) expression in peri-implantation mouse embryos.","date":"2006","source":"Journal of assisted reproduction and genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16622802","citation_count":27,"is_preprint":false},{"pmid":"24703506","id":"PMC_24703506","title":"Heparin-binding EGF-like growth factor (HB-EGF) promotes cell migration and adhesion via focal adhesion kinase.","date":"2014","source":"The Journal of surgical research","url":"https://pubmed.ncbi.nlm.nih.gov/24703506","citation_count":26,"is_preprint":false},{"pmid":"21244855","id":"PMC_21244855","title":"HB-EGF induces cardiomyocyte hypertrophy via an ERK5-MEF2A-COX2 signaling pathway.","date":"2011","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/21244855","citation_count":26,"is_preprint":false},{"pmid":"18281561","id":"PMC_18281561","title":"Preclinical toxicity, toxicokinetics, and antitumoral efficacy studies of DTS-201, a tumor-selective peptidic prodrug of doxorubicin.","date":"2008","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/18281561","citation_count":25,"is_preprint":false},{"pmid":"22641673","id":"PMC_22641673","title":"Regulatory mechanisms of the HB-EGF autocrine loop in inflammation, homeostasis, development and cancer.","date":"2012","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/22641673","citation_count":25,"is_preprint":false},{"pmid":"12028437","id":"PMC_12028437","title":"Role of membrane-bound heparin-binding epidermal growth factor-like growth factor (HB-EGF) in renal epithelial cell branching.","date":"2002","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/12028437","citation_count":25,"is_preprint":false},{"pmid":"19244405","id":"PMC_19244405","title":"Soluble HB-EGF induces epithelial-to-mesenchymal transition in inner medullary collecting duct cells by upregulating Snail-2.","date":"2009","source":"American journal of physiology. Renal physiology","url":"https://pubmed.ncbi.nlm.nih.gov/19244405","citation_count":25,"is_preprint":false},{"pmid":"26637193","id":"PMC_26637193","title":"NMDA receptor agonists reverse impaired psychomotor and cognitive functions associated with hippocampal Hbegf-deficiency in mice.","date":"2015","source":"Molecular brain","url":"https://pubmed.ncbi.nlm.nih.gov/26637193","citation_count":24,"is_preprint":false},{"pmid":"33973871","id":"PMC_33973871","title":"CircRNA_30032 promotes renal fibrosis in UUO model mice via miRNA-96-5p/HBEGF/KRAS axis.","date":"2021","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/33973871","citation_count":24,"is_preprint":false},{"pmid":"22718294","id":"PMC_22718294","title":"Effects of CRM197, a specific inhibitor of HB-EGF, in oral cancer.","date":"2012","source":"Medical molecular morphology","url":"https://pubmed.ncbi.nlm.nih.gov/22718294","citation_count":24,"is_preprint":false},{"pmid":"9724101","id":"PMC_9724101","title":"Targeting tumor cells via EGF receptors: selective toxicity of an HBEGF-toxin fusion protein.","date":"1998","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/9724101","citation_count":24,"is_preprint":false},{"pmid":"24038577","id":"PMC_24038577","title":"Roles for HB-EGF and CD9 in multiple sclerosis.","date":"2013","source":"Glia","url":"https://pubmed.ncbi.nlm.nih.gov/24038577","citation_count":24,"is_preprint":false},{"pmid":"30542468","id":"PMC_30542468","title":"Correlation of IL-1 and HB-EGF with endometrial receptivity.","date":"2018","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/30542468","citation_count":24,"is_preprint":false},{"pmid":"26477568","id":"PMC_26477568","title":"Targeting autocrine HB-EGF signaling with specific ADAM12 inhibition using recombinant ADAM12 prodomain.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26477568","citation_count":23,"is_preprint":false},{"pmid":"26440329","id":"PMC_26440329","title":"Role of G protein-coupled receptors (GPCR), matrix metalloproteinases 2 and 9 (MMP2 and MMP9), heparin-binding epidermal growth factor-like growth factor (hbEGF), epidermal growth factor receptor (EGFR), erbB2, and insulin-like growth factor 1 receptor (IGF-1R) in trenbolone acetate-stimulated bovine satellite cell proliferation.","date":"2015","source":"Journal of animal science","url":"https://pubmed.ncbi.nlm.nih.gov/26440329","citation_count":22,"is_preprint":false},{"pmid":"29191924","id":"PMC_29191924","title":"Both Autocrine Signaling and Paracrine Signaling of HB-EGF Enhance Ocular Neovascularization.","date":"2017","source":"Arteriosclerosis, thrombosis, and vascular biology","url":"https://pubmed.ncbi.nlm.nih.gov/29191924","citation_count":22,"is_preprint":false},{"pmid":"21413023","id":"PMC_21413023","title":"HB-EGF synthesis and release induced by cholesterol depletion of human epidermal keratinocytes is controlled by extracellular ATP and involves both p38 and ERK1/2 signaling pathways.","date":"2011","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/21413023","citation_count":22,"is_preprint":false},{"pmid":"18929421","id":"PMC_18929421","title":"HB-EGF is a paracrine growth stimulator for early tumor prestages in inflammation-associated hepatocarcinogenesis.","date":"2008","source":"Journal of hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/18929421","citation_count":21,"is_preprint":false},{"pmid":"32695675","id":"PMC_32695675","title":"HB-EGF Activates the EGFR/HIF-1α Pathway to Induce Proliferation of Arsenic-Transformed Cells and Tumor Growth.","date":"2020","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/32695675","citation_count":21,"is_preprint":false},{"pmid":"17822789","id":"PMC_17822789","title":"HB-EGF induces delayed STAT3 activation via NF-kappaB mediated IL-6 secretion in vascular smooth muscle cell.","date":"2007","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/17822789","citation_count":21,"is_preprint":false},{"pmid":"33009641","id":"PMC_33009641","title":"APP Binds to the EGFR Ligands HB-EGF and EGF, Acting Synergistically with EGF to Promote ERK Signaling and Neuritogenesis.","date":"2020","source":"Molecular neurobiology","url":"https://pubmed.ncbi.nlm.nih.gov/33009641","citation_count":21,"is_preprint":false},{"pmid":"28368403","id":"PMC_28368403","title":"HBEGF promotes gliomagenesis in the context of Ink4a/Arf and Pten loss.","date":"2017","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/28368403","citation_count":20,"is_preprint":false},{"pmid":"37525152","id":"PMC_37525152","title":"Exosomal hsa_circ_000200 as a potential biomarker and metastasis enhancer of gastric cancer via miR-4659a/b-3p/HBEGF axis.","date":"2023","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/37525152","citation_count":20,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":57359,"output_tokens":10923,"usd":0.167961,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":21547,"output_tokens":6222,"usd":0.131642,"stage2_stop_reason":"end_turn"},"total_usd":0.299603,"stage1_batch_id":"msgbatch_01DUPACZi7AAWwuZiUjseQF4","stage2_batch_id":"msgbatch_018yt7chE3f48PEFtDwtj6AE","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1995,\n      \"finding\": \"Membrane-anchored proHB-EGF forms a complex with the tetraspanin DRAP27/CD9 and integrin α3β1 at cell-cell contact sites, as demonstrated by co-immunoprecipitation, chemical cross-linking, and double-immunofluorescence colocalization in Vero, A431, and MG63 cells.\",\n      \"method\": \"Co-immunoprecipitation, chemical cross-linking, double-immunofluorescence microscopy\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP confirmed by chemical cross-linking and colocalization, multiple cell lines tested\",\n      \"pmids\": [\"7790364\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"CD9, but not CD81 or CD82, upregulates the juxtacrine mitogenic activity of transmembrane proHB-EGF as a diphtheria toxin receptor; chimeric CD9/CD81 molecules mapped this activity to the second half of CD9 (large extracellular loop, fourth TM domain, and C-terminal cytoplasmic domain).\",\n      \"method\": \"Cotransfection of chimeric CD9/CD81 constructs in murine LM cells, diphtheria toxin receptor functional assay\",\n      \"journal\": \"Cellular immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain-mapping by chimeric constructs with functional readout, single lab\",\n      \"pmids\": [\"9514697\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Estrogen-induced ERK1/2 activation requires GPR30-dependent transactivation of the EGF receptor via release of HB-EGF; this was blocked by neutralizing HB-EGF antibodies or by down-modulating HB-EGF from the cell surface with the diphtheria toxin mutant CRM-197, and required Src-related tyrosine kinase activity and Shc-pY317 phosphorylation.\",\n      \"method\": \"Neutralizing antibody treatment, CRM-197 cell-surface depletion, EGFR kinase inhibition, GPR30 cDNA transfection, immunoblot for pERK\",\n      \"journal\": \"Molecular endocrinology (Baltimore, Md.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal inhibitors (antibody, CRM-197, kinase inhibitor, dominant-negative transfection) converging on same conclusion\",\n      \"pmids\": [\"11043579\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"ADAM12 is the metalloprotease responsible for shedding HB-EGF in cardiomyocytes in response to GPCR agonists; dominant-negative ADAM12 expression abrogated shedding-dependent EGFR transactivation and cardiac hypertrophy, and the metalloprotease inhibitor KB-R7785 bound directly to ADAM12.\",\n      \"method\": \"ADAM12 cloning, dominant-negative expression, metalloprotease inhibitor (KB-R7785) direct binding, in vivo cardiac hypertrophy model\",\n      \"journal\": \"Nature medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — dominant-negative genetic approach plus pharmacological inhibitor with direct binding data, replicated in vivo\",\n      \"pmids\": [\"11786904\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"HB-EGF shedding via metalloprotease (MMP) and subsequent EGFR transactivation is required for Angiotensin II-induced fibronectin expression in mesangial cells; ALP-tagged HB-EGF chimera directly measured rapid HB-EGF release (~4-fold within 2 min of Ang II), blocked by PKC inhibitors or MMP inhibitor batimastat.\",\n      \"method\": \"HB-EGF-alkaline phosphatase chimera shedding assay, neutralizing anti-HB-EGF antibody, EGFR inhibitor AG1478, heparin blocking, batimastat MMP inhibition\",\n      \"journal\": \"Kidney international\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — direct measurement of shedding with reporter chimera, multiple pharmacological blockers, single lab\",\n      \"pmids\": [\"11737589\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Helicobacter pylori triggers EGFR tyrosine phosphorylation via HB-EGF gene induction and metalloprotease-dependent ectodomain shedding (the 'triple membrane passing signal'); this signaling also enhances IL-8 secretion.\",\n      \"method\": \"EGFR phosphorylation assay, HB-EGF gene expression analysis, metalloprotease inhibition, EGFR and MEK1 inhibitors\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple pathway inhibitors, pharmacological validation, single lab\",\n      \"pmids\": [\"12099696\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"TGF-β mediates fibronectin expression in mesangial cells via PC-PLC/PKC-dependent metalloprotease-mediated HB-EGF release and subsequent EGFR transactivation; Smad2 phosphorylation by TGF-β was unaffected by EGFR inhibition, and HB-EGF did not activate Smad2, indicating a parallel pathway.\",\n      \"method\": \"HB-EGF-alkaline phosphatase chimera shedding assay, neutralizing anti-HB-EGF antibody, AG1478, PKC inhibitors, batimastat, FN mRNA quantification\",\n      \"journal\": \"Kidney international\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — direct shedding reporter, multiple inhibitors, epistasis between Smad2 and EGFR pathways established, single lab\",\n      \"pmids\": [\"12164862\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"HB-EGF null mice exhibit enlarged, malformed cardiac valves due to abnormal mesenchymal cell proliferation during remodeling, associated with increased activated Smad1/5/8; similar defects occur in EGFR- and TACE-null mice, establishing that TACE-derived soluble HB-EGF activates EGFR to suppress BMP signaling during cardiac valvulogenesis.\",\n      \"method\": \"Homologous recombination knockout mice, genetic epistasis (HB-EGF−/−, EGFR−/−, TACE−/−), immunohistochemistry for pSmad1/5/8, histological analysis\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — three independent genetic knockouts show convergent cardiac valve phenotype, epistasis established\",\n      \"pmids\": [\"12773386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Wound-induced MMP-dependent ectodomain shedding of HB-EGF acts as an autocrine/paracrine EGFR ligand in corneal epithelial wound closure; wound-induced EGFR and ERK phosphorylation were blocked by CRM-197 (HB-EGF antagonist), HB-EGF function-blocking antibodies, and MMP inhibitor GM6001, with direct HB-EGF shedding demonstrated by HB-EGF-AP reporter.\",\n      \"method\": \"HB-EGF-alkaline phosphatase shedding assay, CRM-197, neutralizing HB-EGF antibodies, GM6001 MMP inhibitor, AG1478 EGFR inhibitor, immunoprecipitation + Western blot\",\n      \"journal\": \"Investigative ophthalmology & visual science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct shedding reporter plus multiple orthogonal inhibitors confirmed in both organ culture and cell culture models\",\n      \"pmids\": [\"14985295\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Soluble HB-EGF, but not pro-HB-EGF (membrane-only form), confers tumorigenic properties including enhanced growth, colony formation, cell migration, cyclin D1 activation, VEGF induction, and MMP-9/MMP-3 upregulation in bladder cancer cells in vitro and in vivo.\",\n      \"method\": \"Tetracycline-regulatable expression system, soft agar colony assay, xenograft nude mouse tumorigenesis, zymography for MMP activity\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — inducible expression with paired s-HB-EGF vs pro-HB-EGF controls, multiple in vitro and in vivo readouts\",\n      \"pmids\": [\"15289334\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Membrane-anchored (noncleavable) HB-EGF promotes cell-matrix and cell-cell interactions and decreases migration and HGF-induced scattering in MDCK cells, while soluble HB-EGF has the opposite effects, demonstrating that the two forms of HB-EGF have distinctly different functional consequences.\",\n      \"method\": \"Stable transfection of proHB-EGF, noncleavable deletion mutant, or soluble HB-EGF in MDCK cells; migration, scattering, tubulogenesis, and adhesion assays\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic approach with matched cell lines expressing distinct forms, multiple orthogonal functional assays, single lab\",\n      \"pmids\": [\"14996914\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"HB-EGF directs stromal cell polyploidy and decidualization via upregulation of cyclin D3; adenoviral antisense delivery of cyclin D3 abrogated HB-EGF-induced polyploidy both in vitro and in vivo.\",\n      \"method\": \"Adenoviral antisense cyclin D3 delivery, BrdU labeling, flow cytometry, in vivo uterine decidualization model\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function via antisense in vitro and in vivo with specific phenotypic readout, single lab\",\n      \"pmids\": [\"14697362\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"HB-EGF promotes eyelid closure by activating EGFR-ERK signaling at the leading edge of migrating epithelium; HB-EGF null and secretion-deficient mutant mice show delayed eyelid closure with reduced actin bundle formation; genetic interaction with hypomorphic EGFR (waved-2) confirmed; TGFα acts synergistically and equally.\",\n      \"method\": \"HB-EGF null mouse KO, secretion-deficient knock-in (HB-uc/uc), EGFR kinase inhibitor in vivo, double null crosses (HB-EGF × TGFα), pEGFR and pERK immunostaining\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple genetic models (KO, knock-in, hypomorph, double null) with converging mechanistic pathway\",\n      \"pmids\": [\"16141218\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"HB-EGF enhances intestinal restitution via PI3K/Akt and MEK/ERK1/2 signaling downstream of ErbB-1; blocking ErbB-1, PI3K/Akt, or MEK/ERK significantly reduced intrinsic and HB-EGF-induced restitution in vitro, and endogenous HB-EGF was shown to be essential for wound-induced ErbB-1 and ERK1/2 activation.\",\n      \"method\": \"Intestinal I/R model in rats, scrape wound assay in vitro, pharmacological inhibitors of ErbB-1/PI3K/MEK, CRM-197 HB-EGF blockade, immunoblot for pAkt and pERK\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo and in vitro models with pathway inhibitors and endogenous HB-EGF neutralization, single lab\",\n      \"pmids\": [\"16083716\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"5-HT2A receptor induces ERK phosphorylation and cellular proliferation via TACE (ADAM17) activation, HB-EGF shedding, and EGFR transactivation in mesangial cells; TACE was co-immunoprecipitated with HB-EGF, and TACE siRNA blocked HB-EGF shedding, ERK phosphorylation, and DNA content increase.\",\n      \"method\": \"TACE siRNA knockdown, neutralizing HB-EGF antibody, EGFR inhibitor AG1478, MMP inhibitors, co-immunoprecipitation of TACE with HB-EGF, DNA content measurement\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — siRNA, pharmacological inhibitors, and Co-IP converging on TACE–HB-EGF complex; multiple orthogonal methods\",\n      \"pmids\": [\"16737974\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"MMP-7 catalyzes release of HB-EGF to mediate bile acid (deoxycholyltaurine)-induced EGFR transactivation and colon cancer cell proliferation; MMP-7 and proHB-EGF co-localize at the cell surface, and specific MMP-7 knockdown (neutralizing antibody or siRNA) attenuated DCT-induced signaling and proliferation.\",\n      \"method\": \"MMP-7 siRNA, neutralizing antibody to MMP-7, CRM-197 HB-EGF inhibitor, EGFR ligand-domain antibody, immunofluorescence colocalization, qRT-PCR for MMP-7\",\n      \"journal\": \"Biochemical pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA and neutralizing antibody plus colocalization evidence, single lab\",\n      \"pmids\": [\"17222808\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"HB-EGF promotes angiogenesis (migration and tube formation) in HUVEC via activation of PI3K, MAPK, and eNOS pathways, in a VEGF-independent manner.\",\n      \"method\": \"Scratch wound migration assay, 2D tube formation assay, pharmacological inhibitors of PI3K, MAPK, eNOS\",\n      \"journal\": \"Growth factors (Chur, Switzerland)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple pathway inhibitors with functional angiogenesis readouts, single lab\",\n      \"pmids\": [\"18092233\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"ERK1/2 mediates ADAM-dependent HB-EGF shedding and EGFR transactivation in corneal epithelial cells in response to wounding and GPCR ligands; ADAM17 was co-immunoprecipitated with active ERK and phosphorylated at serine residues in an ERK-dependent manner.\",\n      \"method\": \"MEK inhibitors PD98059/U0126, HB-EGF-AP shedding assay, co-immunoprecipitation of ERK with ADAM17, immunoprecipitation + Western blot for serine phosphorylation of ADAM17\",\n      \"journal\": \"Investigative ophthalmology & visual science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP of ADAM17-ERK complex plus phosphorylation evidence plus direct shedding reporter, multiple stimuli tested\",\n      \"pmids\": [\"18658095\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"HB-EGF stimulates eNOS expression and NO production in endothelial cells via a PI3K-dependent pathway; eNOS siRNA and eNOS-specific inhibitors abolished HB-EGF-induced HUVEC migration and angiogenesis in vitro and in vivo (Matrigel plug assay).\",\n      \"method\": \"eNOS siRNA knockdown, L-NAME/L-NIO pharmacological eNOS inhibitors, PI3K inhibitor, Matrigel plug in vivo angiogenesis, Western blot for eNOS protein/phosphorylation\",\n      \"journal\": \"Growth factors (Chur, Switzerland)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA plus pharmacological inhibitors plus in vivo assay, single lab\",\n      \"pmids\": [\"18925469\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"The heparin-binding domain of transmembrane proHB-EGF mediates localization to cell-cell contact sites via trans interaction with extracellular heparan sulfate proteoglycans (HSPGs); disruption of this interaction increases proteolytic release of soluble ligand and switches cell behavior from juxtacrine growth inhibition to autocrine proliferation.\",\n      \"method\": \"Independent tracking of extracellular EGF domain and cytoplasmic C-terminus with new labeling method, heparin treatment, HSPG disruption experiments, wound-closure assay, proliferation assays\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — novel dual-domain tracking method with functional readouts, single lab\",\n      \"pmids\": [\"20530570\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Soluble HB-EGF induces epithelial-to-mesenchymal transition (EMT) in renal collecting duct cells by upregulating the E-cadherin transcriptional repressor Snail-2; stable Snail-2 knockdown restored epithelial markers but not anchorage-independent growth.\",\n      \"method\": \"Stable overexpression of sHB-EGF, qRT-PCR and luciferase reporter for E-cadherin transcription, stable shRNA knockdown of Snail-2, Western blot for epithelial/mesenchymal markers\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain-of-function and epistasis via Snail-2 knockdown, single lab\",\n      \"pmids\": [\"19244405\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Mechanical stretch promotes fetal type II epithelial cell differentiation via ectodomain shedding of HB-EGF and TGF-α; 5% cyclic stretch of E19 fetal cells transfected with AP-tagged HB-EGF directly measured HB-EGF release, which was not enhanced by fibroblast co-culture.\",\n      \"method\": \"AP-tagged HB-EGF shedding assay under cyclic stretch, SP-B/C mRNA and protein measurement, recombinant HB-EGF/TGFα/amphiregulin/BTC/EPR comparison\",\n      \"journal\": \"The Journal of physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — direct shedding reporter under defined mechanical conditions, single lab\",\n      \"pmids\": [\"19237431\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Integrin/FAK signaling masks HB-EGF's growth-promoting activity in standard 2D monolayer culture; reducing integrin β1 or FAK by antibody or genetic knockout revealed HB-EGF-dependent cell growth; HB-EGF-driven growth was clearly demonstrated in 3D culture or when integrin signaling was attenuated.\",\n      \"method\": \"Integrin β1 antibody blocking, FAK knockout, 3D culture system, xenograft tumor growth comparison\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO and antibody approaches in multiple culture contexts, single lab\",\n      \"pmids\": [\"19887590\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"HB-EGF induces cardiomyocyte hypertrophy via an EGFR-MEK5-ERK5-MEF2A-COX-2 signaling pathway; MEK5 siRNA reduced HB-EGF-induced cell size and ANF mRNA expression, MEF2A siRNA attenuated COX-2 induction, and COX-2 inhibitor rofecoxib reduced ANF expression.\",\n      \"method\": \"MEK5 siRNA, MEF2A siRNA, COX-2 inhibitor rofecoxib, AG1478 EGFR inhibitor, [3H]-leucine incorporation, ANF mRNA measurement\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple siRNA knockdowns defining sequential pathway, single lab\",\n      \"pmids\": [\"21244855\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"LIV-1 overexpression promotes EMT in prostate cancer cells via MMP2/MMP9-mediated shedding of HB-EGF, causing constitutive EGFR phosphorylation and downstream ERK signaling.\",\n      \"method\": \"LIV-1 overexpression, zymography for MMP2/MMP9 activity, EGFR phosphorylation by Western blot, ERK activation assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, gain-of-function with pathway readouts but limited mechanistic dissection of HB-EGF specifically\",\n      \"pmids\": [\"22110740\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"miR-1207-5p directly regulates HBEGF expression in podocytes via its 3'UTR; a miRSNP C1936T at position 2 of the seed region abolishes this regulation, as shown by luciferase reporter assay and western blot with miRNA mimics.\",\n      \"method\": \"Luciferase reporter assay with 3'UTR construct, miRNA mimic transfection, Western blot for HBEGF protein in podocytes\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct reporter and protein-level validation of miRNA-target interaction with SNP functional analysis, single lab\",\n      \"pmids\": [\"22319602\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Hypoxia increases ADAM12 levels and activity in a Notch signaling-dependent manner, leading to increased HB-EGF ectodomain shedding; released HB-EGF induces invadopodium formation in cancer cells, linking Notch and EGFR pathways in promoting invasion.\",\n      \"method\": \"Hypoxia treatment, Notch pathway manipulation, ADAM12 activity assay, HB-EGF shedding measurement, invadopodium formation assay\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and pharmacological Notch manipulation linked to ADAM12-HB-EGF-invadopodia axis, single lab\",\n      \"pmids\": [\"23589494\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Autocrine HB-EGF expression in breast cancer cells enhances invadopodium formation and function via EGFR signaling, and upregulates MMP2 and MMP9; HB-EGF inhibition rapidly decreased invadopodia, demonstrating direct EGFR-dependent mechanism.\",\n      \"method\": \"HB-EGF overexpression, EGFR inhibition, in vivo intravasation assay, MMP2/9 expression analysis, invadopodium formation assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain-of-function with EGFR inhibitor rescue and in vivo confirmation, single lab\",\n      \"pmids\": [\"24013225\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"miR-96 promotes osteogenic differentiation by suppressing HB-EGF post-transcriptionally via binding to the 3'UTR of HB-EGF mRNA, thereby reducing EGFR and ERK1/AKT phosphorylation downstream.\",\n      \"method\": \"miR-96 overexpression in MC3T3-E1 cells, 3'UTR binding assay, Western blot for HB-EGF/pEGFR/pERK1/pAKT, osteogenic differentiation markers\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct 3'UTR binding with downstream pathway validation, single lab\",\n      \"pmids\": [\"25451232\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"HB-EGF promotes intestinal epithelial cell migration and adhesion via FAK phosphorylation; HB-EGF increased p-FAK expression and induced FAK redistribution and actin reorganization, and FAK inhibitor reversed HB-EGF-induced migration and adhesion.\",\n      \"method\": \"Scrape wound healing assay, FAK inhibitor 14, immunofluorescence for p-FAK redistribution, Western blot, fibronectin adhesion assay\",\n      \"journal\": \"The Journal of surgical research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological FAK inhibition with multiple functional readouts, single lab\",\n      \"pmids\": [\"24703506\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"In head and neck squamous cell carcinoma, Abl kinases negatively regulate invadopodia through suppression of an HB-EGF autocrine loop; imatinib stimulated HB-EGF shedding from HNSCC cells, and soluble HB-EGF enhanced invadopodia ECM degradation via an EGFR-Src-cortactin cascade.\",\n      \"method\": \"Imatinib treatment, HB-EGF shedding measurement, Abl/Arg siRNA, invadopodium formation and ECM degradation assays, EGFR/Src inhibitors\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA and pharmacological inhibitors establishing pathway context, single lab\",\n      \"pmids\": [\"23146907\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Recombinant ADAM12 prodomain (PA12) selectively inhibits ADAM12 but not ADAM10 or ADAM17, reducing HB-EGF shedding and cellular migration in endometriotic cells; computational modeling predicted that high-affinity ligands like HB-EGF are more difficult to target with decoy antibodies than low-affinity ligands.\",\n      \"method\": \"Recombinant ADAM12 prodomain protein, HB-EGF shedding assay, selectivity profiling against ADAM10/17, cell migration assay, computational modeling\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro biochemical inhibition with selectivity profiling plus functional validation, single lab\",\n      \"pmids\": [\"26477568\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"HBEGF can initiate glioblastoma in mice with Ink4a/Arf and Pten loss; HBEGF stimulation activates both EGFR and the RTK Axl, and EGFR is required for Axl activation; silencing HBEGF in vivo caused tumor regression.\",\n      \"method\": \"In vivo mouse GBM model, EGFR/Axl conditional deletion, in vivo HBEGF silencing with survival analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic model with epistasis between EGFR and Axl, single lab\",\n      \"pmids\": [\"28368403\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"HB-EGF gene expression in β-cells is increased by glucose in a ChREBP-dependent manner; ChREBP binding sites near the HB-EGF gene were identified by ChIP; HB-EGF knockdown in rat islets blocked glucose-induced β-cell proliferation ex vivo and in vivo; Src family kinase inhibition abrogated glucose-induced proliferation, implicating Src in HB-EGF processing.\",\n      \"method\": \"HB-EGF siRNA knockdown in isolated islets, transplanted glucose-infused rats, ChREBP ChIP, EGFR and HB-EGF inhibitors, Src kinase inhibitors, β-cell proliferation assay\",\n      \"journal\": \"Diabetes\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP for transcriptional regulation, in vitro and in vivo KD with functional rescue, multiple inhibitors, single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"31882563\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"BHLHE40 induces HBEGF transcription by blocking DNA binding of HDAC1 and HDAC2, as shown by ChIP and Co-IP assays; HBEGF is then secreted through exosomes to promote cell survival and migration.\",\n      \"method\": \"ChIP assay, Co-IP of BHLHE40 with HDAC1/2, exosome analysis, HBEGF knockdown, migration/survival assays, xenograft and experimental metastasis models\",\n      \"journal\": \"Breast cancer research : BCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and Co-IP identify transcriptional mechanism, functional validation in vivo, single lab\",\n      \"pmids\": [\"30285805\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Myeloid cell-derived HB-EGF induces epithelial EGFR nuclear translocation and methylation of histone H4 to facilitate DNA damage repair in pancreatic acinar cells; conditional knockout of HB-EGF in myeloid cells (LysM-Cre) delayed recovery from pancreatitis with impaired DNA repair and increased apoptosis.\",\n      \"method\": \"Conditional myeloid-specific Hbegf KO (LysM-Cre), myeloid cell depletion (CD11b-DTR), EGFR conditional KO in pancreatic epithelium, H4 methylation immunostaining, DNA damage assays, cell proliferation/apoptosis measurements\",\n      \"journal\": \"Cellular and molecular gastroenterology and hepatology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple conditional KO models converging on HB-EGF/EGFR/H4 methylation axis, in vitro mechanistic validation\",\n      \"pmids\": [\"31125624\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"s-HBEGF released from sorafenib-treated vascular endothelial cells activates EGFR on keratinocytes and promotes JNK2-mediated SIRT1 stabilization, driving hyper-keratinization; HBEGF neutralization, SIRT1 knockdown, or nicotinamide (SIRT1 inhibitor) all reduced sorafenib-induced hand-foot skin reaction in mouse models.\",\n      \"method\": \"In vivo mouse HFSR model, s-HBEGF administration, HBEGF neutralization antibody, SIRT1 siRNA knockdown, nicotinamide treatment, pJNK2 and SIRT1 Western blot, preliminary clinical study\",\n      \"journal\": \"Cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo model with genetic and pharmacological interventions defining s-HBEGF→EGFR→JNK2→SIRT1 pathway, single lab\",\n      \"pmids\": [\"32296111\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"HBEGF+ inflammatory macrophages in rheumatoid arthritis promote fibroblast invasiveness in an EGF receptor-dependent manner; this cross-talk is shaped by resident fibroblasts and TNF.\",\n      \"method\": \"Single-cell RNA sequencing, ex vivo synovial tissue assay with EGFR pathway inhibition, functional invasiveness assays\",\n      \"journal\": \"Science translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — scRNA-seq identification plus functional ex vivo validation with EGFR-dependence shown, single study\",\n      \"pmids\": [\"31068444\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TMPRSS4 increases both transcriptional/translational levels of HB-EGF precursor and promotes its proteolytic cleavage by enhancing MMP9 expression via EGFR/Akt/mTOR/HIF-1α signaling; HB-EGF in turn promotes HCC proliferation and invasion via EGFR/PI3K/Akt pathway.\",\n      \"method\": \"TMPRSS4 overexpression/knockdown, MMP9 activity assay, EGFR/Akt/mTOR pathway inhibitors, CRM-197 HB-EGF inhibitor, in vivo xenograft model\",\n      \"journal\": \"Hepatology (Baltimore, Md.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain/loss-of-function with pathway inhibitors in vitro and in vivo, single lab\",\n      \"pmids\": [\"31867749\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"APP interacts with pro-HB-EGF (identified in yeast two-hybrid screen and confirmed by co-immunoprecipitation); APP and EGF synergistically activate ERK signaling and promote neuritogenesis via EGFR, as shown by EGFR inhibitor PD 168393 abrogation.\",\n      \"method\": \"Yeast two-hybrid screen, co-immunoprecipitation, EGFR inhibitor, immunofluorescence neurite analysis, Western blot for pERK\",\n      \"journal\": \"Molecular neurobiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast two-hybrid plus Co-IP confirms interaction; functional EGFR-dependence established by inhibitor, single lab\",\n      \"pmids\": [\"33009641\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Group 3 innate lymphoid cells (ILC3s) produce HB-EGF in response to prostaglandin E2/EP2 receptor engagement; ILC3-derived HB-EGF protects intestinal epithelium from TNF-induced cell death independently of IL-22; mice lacking ILC3-derived HB-EGF showed increased susceptibility to TNF-mediated epithelial death and experimental intestinal inflammation.\",\n      \"method\": \"Conditional HB-EGF KO in ILC3s (Hbegf f/f × ILC3-Cre), PGE2 stimulation assays, TNF-induced intestinal injury model, in vitro epithelial protection assays\",\n      \"journal\": \"Nature immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cell-type-specific conditional KO with in vivo and in vitro mechanistic dissection, multiple orthogonal approaches\",\n      \"pmids\": [\"35102343\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Combined TNF and HBEGF treatment in human retinal organoids induces photoreceptor neurodegeneration via a previously unknown cell extrusion mechanism; pharmacological inhibitors of PIEZO1 (mechanosensor), MAPK, and actomyosin each prevented pathogenesis, and a PIEZO1 activator alone induced photoreceptor extrusion.\",\n      \"method\": \"Human retinal organoid system, combined HBEGF+TNF treatment, PIEZO1 pharmacological activation and inhibition, MAPK/actomyosin inhibitors, live imaging, transcriptome analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — organoid model with pharmacological pathway dissection revealing novel mechanism, single study\",\n      \"pmids\": [\"36261438\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Hb-egfa produced by ependymal cells is required for spinal cord regeneration in zebrafish; hb-egfa mutants show defective axon crossing, tissue bridging, and swim recovery after transection; local recombinant HB-EGF delivery alters ependymal cell cycling and enhances functional regeneration; a tissue regeneration enhancer element (TREE) linked to hb-egfa drives gene expression specifically at spinal cord injuries.\",\n      \"method\": \"hb-egfa mutant zebrafish, recombinant HB-EGF delivery, epigenetic profiling for TREE identification, AAV-based enhancer-driven delivery in neonatal mouse spinal cord crush, BrdU/EdU cell cycling assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic mutant plus recombinant protein rescue plus epigenetic TREE characterization with cross-species validation in mouse\",\n      \"pmids\": [\"37567873\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Astrocyte-derived HB-EGF limits autoimmune CNS pathology; hypoxic conditions rapidly upregulate HB-EGF in astrocytes, while pro-inflammatory conditions suppress HB-EGF signaling through epigenetic modifications; intranasal HB-EGF administration attenuated disease in a mouse EAE model.\",\n      \"method\": \"Proteomic analysis of CSF, in vitro cell type studies, epigenetic modification analysis of HB-EGF locus, intranasal HB-EGF delivery in EAE model\",\n      \"journal\": \"Nature immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro mechanistic studies plus in vivo therapeutic validation, but epigenetic details are limited in the abstract, single study\",\n      \"pmids\": [\"38409259\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"HBEGF encodes a dual-form growth factor (membrane-anchored proHB-EGF and shed soluble HB-EGF) that binds and activates EGFR/ErbB4; its ectodomain is shed by ADAM10, ADAM12, or ADAM17 (TACE) in response to diverse stimuli including GPCR agonists, mechanical stress, wounding, and hypoxia, generating a paracrine/autocrine signal that activates ERK, PI3K/Akt, and downstream transcriptional programs to drive cardiac valve morphogenesis, epithelial wound closure, β-cell proliferation, intestinal repair, neurogenesis, and cancer invasion, while membrane-retained proHB-EGF engages in juxtacrine signaling by complexing with CD9 and integrin α3β1 at cell-cell contacts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"HBEGF encodes a dual-form EGFR ligand that exists as a membrane-anchored precursor (proHB-EGF) and a proteolytically shed soluble form, and these two forms drive opposing cellular programs across morphogenesis, tissue repair, and cancer [#9, #10]. Membrane-anchored proHB-EGF localizes to cell-cell contacts in a complex with the tetraspanin CD9 and integrin \\u03b13\\u03b21, where CD9 upregulates its juxtacrine activity, and its heparin-binding domain anchors it at contact sites through trans interaction with heparan sulfate proteoglycans; disrupting this interaction promotes proteolytic release and switches behavior from juxtacrine growth inhibition to autocrine proliferation [#0, #1, #19]. Ectodomain shedding is executed by metalloproteases\\u2014ADAM12 in cardiomyocytes, ADAM17/TACE downstream of GPCRs, and MMP-7/MMP2/MMP9 in epithelial and cancer contexts\\u2014and is triggered by GPCR agonists (estrogen via GPR30, angiotensin II, 5-HT2A), mechanical stretch, wounding, and hypoxia/Notch signaling, with ERK1/2 feeding back to phosphorylate and activate ADAM17 [#2, #3, #14, #17, #21, #26]. Soluble HB-EGF transactivates EGFR to engage ERK and PI3K/Akt cascades, driving cardiac valve morphogenesis through suppression of BMP/Smad signaling, eyelid and corneal epithelial closure, intestinal restitution, glucose-stimulated \\u03b2-cell proliferation via ChREBP-driven transcription, and angiogenesis through a PI3K/eNOS axis [#7, #8, #12, #13, #16, #18, #33]. In disease, soluble HB-EGF promotes tumorigenesis, EMT via Snail-2, and EGFR-dependent invadopodium formation, can initiate glioblastoma through coordinated EGFR and Axl activation, and mediates inflammatory tissue protection and regeneration including ILC3- and astrocyte-derived epithelial/CNS protection and zebrafish spinal cord regeneration [#9, #20, #27, #32, #40, #42, #43]. HBEGF expression is controlled transcriptionally (ChREBP, BHLHE40) and post-transcriptionally (miR-1207-5p, miR-96 acting on its 3'UTR) [#25, #28, #33, #34].\",\n  \"teleology\": [\n    {\n      \"year\": 1995,\n      \"claim\": \"Established that the membrane-anchored precursor is not merely a latent ligand reservoir but a structured signaling unit at cell-cell contacts, defining its juxtacrine context.\",\n      \"evidence\": \"Co-IP, chemical cross-linking, and colocalization of proHB-EGF with CD9 and integrin \\u03b13\\u03b21 in Vero/A431/MG63 cells\",\n      \"pmids\": [\"7790364\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of the CD9/integrin complex for signaling not resolved here\", \"Stoichiometry and structure of the complex undefined\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Mapped which tetraspanin partner and which protein domains confer juxtacrine mitogenic enhancement, distinguishing CD9 from related tetraspanins.\",\n      \"evidence\": \"Chimeric CD9/CD81 cotransfection in LM cells with diphtheria toxin receptor functional readout\",\n      \"pmids\": [\"9514697\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab\", \"Mechanism by which the CD9 C-terminal region enhances activity unknown\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Defined HB-EGF shedding as the mechanistic link in GPCR-to-EGFR transactivation across multiple receptors, converting diverse extracellular cues into EGFR signaling.\",\n      \"evidence\": \"Estrogen/GPR30, angiotensin II, and TGF-\\u03b2 systems using HB-EGF-AP shedding reporters, CRM-197, neutralizing antibodies, and EGFR/MMP/PKC inhibitors\",\n      \"pmids\": [\"11043579\", \"11737589\", \"12164862\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific protease assignments varied by stimulus\", \"In vivo relevance of individual GPCR-driven shedding events not fully established\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Identified ADAM12 as the physiological sheddase coupling GPCR agonists to EGFR transactivation in cardiomyocyte hypertrophy, providing the first defined protease-ligand-receptor cardiac axis.\",\n      \"evidence\": \"Dominant-negative ADAM12, direct KB-R7785 binding, in vivo cardiac hypertrophy model\",\n      \"pmids\": [\"11786904\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether ADAM12 is the sole sheddase in heart not excluded\", \"Direct demonstration of HB-EGF as the only relevant ADAM12 substrate lacking\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Demonstrated a developmental requirement: soluble HB-EGF activates EGFR to suppress BMP/Smad signaling during cardiac valvulogenesis, showing the genetic non-redundancy of the ligand.\",\n      \"evidence\": \"HB-EGF\\u2212/\\u2212, EGFR\\u2212/\\u2212, and TACE\\u2212/\\u2212 knockout mice with convergent valve phenotype and pSmad1/5/8 staining\",\n      \"pmids\": [\"12773386\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct biochemical link between EGFR activation and Smad suppression not delineated\", \"Cell-type source of HB-EGF in valve not pinpointed\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Distinguished the opposing functions of the two HB-EGF forms\\u2014soluble form is tumorigenic and pro-migratory while membrane form promotes adhesion and inhibits migration\\u2014establishing form-specific biology.\",\n      \"evidence\": \"Inducible and stable expression of soluble vs noncleavable proHB-EGF in bladder cancer and MDCK cells; xenograft, colony, migration, and adhesion assays\",\n      \"pmids\": [\"15289334\", \"14996914\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of membrane-form growth inhibition not fully resolved\", \"Single-cell-line dependence of some readouts\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Showed HB-EGF drives epithelial wound closure and restitution through EGFR-ERK and PI3K/Akt, defining the effector cascades for tissue repair.\",\n      \"evidence\": \"Corneal/eyelid wound models with HB-EGF null and secretion-deficient mice, EGFR hypomorphs, and intestinal I/R with pathway inhibitors\",\n      \"pmids\": [\"14985295\", \"16141218\", \"16083716\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative contribution of ERK vs PI3K branches tissue-dependent\", \"Identity of sheddase in each wound context not uniformly defined\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Extended the sheddase repertoire to TACE/ADAM17 and MMP-7 for specific GPCR and bile-acid stimuli, showing protease usage is stimulus- and tissue-specific.\",\n      \"evidence\": \"TACE siRNA and Co-IP with HB-EGF in mesangial cells; MMP-7 siRNA/neutralizing antibody and colocalization in colon cancer cells\",\n      \"pmids\": [\"16737974\", \"17222808\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab per system\", \"Direct cleavage by these proteases not reconstituted biochemically\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Revealed a feedback loop in which ERK1/2 phosphorylates and activates ADAM17, amplifying HB-EGF shedding and EGFR transactivation.\",\n      \"evidence\": \"MEK inhibitors, HB-EGF-AP shedding assay, ERK-ADAM17 Co-IP and serine phosphorylation in corneal epithelial cells\",\n      \"pmids\": [\"18658095\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphosite mapping incomplete\", \"Generality beyond corneal epithelium untested\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined how proHB-EGF is retained at contacts and how its release is gated, linking heparan sulfate binding to the juxtacrine-to-autocrine switch.\",\n      \"evidence\": \"Dual-domain tracking, heparin/HSPG disruption, and proliferation/wound assays; integrin/FAK masking of HB-EGF growth activity in 2D vs 3D\",\n      \"pmids\": [\"20530570\", \"19887590\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab\", \"In vivo relevance of HSPG-mediated retention not demonstrated\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Connected HB-EGF to EMT and mechanotransduction-driven differentiation, broadening its program beyond proliferation.\",\n      \"evidence\": \"sHB-EGF overexpression with Snail-2 knockdown in renal cells; AP-tagged HB-EGF shedding under cyclic stretch in fetal type II cells\",\n      \"pmids\": [\"19244405\", \"19237431\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab\", \"Receptor-level coupling to Snail-2 induction not mapped\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Placed HB-EGF at the center of cancer invasion machinery, linking hypoxia/Notch-induced ADAM12 shedding and autocrine EGFR signaling to invadopodium formation.\",\n      \"evidence\": \"Hypoxia/Notch manipulation, ADAM12 activity, and invadopodium assays in cancer cells; autocrine HB-EGF overexpression and EGFR inhibition with in vivo intravasation\",\n      \"pmids\": [\"23589494\", \"24013225\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab per study\", \"Quantitative contribution to metastasis in vivo limited\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Profiled selective ADAM12 inhibition and Abl-mediated negative regulation of the HB-EGF autocrine loop, identifying therapeutic targeting and regulatory nodes.\",\n      \"evidence\": \"Recombinant ADAM12 prodomain selectivity profiling and migration assay; imatinib-induced HB-EGF shedding and EGFR-Src-cortactin cascade in HNSCC\",\n      \"pmids\": [\"26477568\", \"23146907\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"High-affinity HB-EGF predicted hard to target by decoys\", \"Single lab per study\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Showed HB-EGF can act as a tumor initiator through coordinated activation of EGFR and the RTK Axl, with EGFR upstream of Axl.\",\n      \"evidence\": \"In vivo GBM model with Ink4a/Arf and Pten loss, conditional EGFR/Axl deletion, in vivo HBEGF silencing\",\n      \"pmids\": [\"28368403\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of EGFR-dependent Axl activation undefined\", \"Single model system\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established the regulatory inputs controlling HBEGF gene expression, both transcriptional (ChREBP, BHLHE40 via HDAC blockade) and post-transcriptional (miR-1207-5p, miR-96), linking nutrient and oncogenic signals to ligand abundance.\",\n      \"evidence\": \"ChREBP and BHLHE40 ChIP/Co-IP with islet and breast cancer functional assays; luciferase 3'UTR reporters and miRNA mimics in podocytes and osteoblasts\",\n      \"pmids\": [\"31882563\", \"30285805\", \"22319602\", \"25451232\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Interplay among these regulators untested\", \"Tissue-specificity of each control mechanism partially defined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified immune- and stroma-derived HB-EGF as a driver of tissue repair and pathology, including myeloid HB-EGF promoting DNA repair via EGFR nuclear translocation and macrophage HB-EGF driving fibroblast invasiveness.\",\n      \"evidence\": \"Conditional myeloid Hbegf KO with H4 methylation/DNA repair assays in pancreatitis; scRNA-seq and ex vivo EGFR-inhibition assays in rheumatoid arthritis synovium\",\n      \"pmids\": [\"31125624\", \"31068444\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of EGFR nuclear translocation incompletely defined\", \"Direct sheddase in immune contexts not specified\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Expanded the receptor and partner network, showing APP physically binds proHB-EGF and synergizes with EGF in neuritogenesis, and that TMPRSS4 amplifies both HB-EGF precursor levels and its cleavage in HCC.\",\n      \"evidence\": \"Yeast two-hybrid and Co-IP for APP-proHB-EGF with EGFR-dependent neurite assays; TMPRSS4 gain/loss with MMP9 and EGFR/PI3K/Akt readouts in HCC\",\n      \"pmids\": [\"33009641\", \"31867749\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional significance of APP interaction in vivo unknown\", \"Single lab per study\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined protective and pathogenic context-dependent roles of HB-EGF in mucosal and neural tissue, from ILC3-derived epithelial protection to PIEZO1-coupled photoreceptor extrusion.\",\n      \"evidence\": \"ILC3-specific Hbegf conditional KO with TNF-injury models; human retinal organoid HBEGF+TNF treatment with PIEZO1/MAPK/actomyosin inhibitors\",\n      \"pmids\": [\"35102343\", \"36261438\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor coupling for ILC3-derived protection not fully mapped\", \"Organoid extrusion mechanism single study\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrated regenerative and immunoprotective HB-EGF functions in the CNS, with ependymal HB-EGF required for spinal cord regeneration and astrocyte-derived HB-EGF limiting autoimmune pathology.\",\n      \"evidence\": \"hb-egfa zebrafish mutants with recombinant rescue and TREE characterization in mouse; CSF proteomics and intranasal HB-EGF in EAE\",\n      \"pmids\": [\"37567873\", \"38409259\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream effector pathways in regeneration not detailed\", \"Epigenetic control of astrocyte HB-EGF only partially characterized\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the choice between juxtacrine proHB-EGF signaling and autocrine/paracrine soluble HB-EGF signaling is integrated at the level of a single cell\\u2014and which sheddase dominates in each physiological context in vivo\\u2014remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model of context-specific sheddase selection in vivo\", \"Structural basis of the proHB-EGF/CD9/integrin complex undefined\", \"Quantitative balance between membrane-form inhibition and soluble-form activation not established\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [2, 7, 8, 9, 13]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [19]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [2, 7, 13, 33]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1, 15, 19]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [9, 8, 34]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 3, 13, 14, 17]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [7, 12, 42]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [9, 27, 32, 38]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [37, 40, 43]}\n    ],\n    \"complexes\": [\"proHB-EGF/CD9/integrin \\u03b13\\u03b21 complex\"],\n    \"partners\": [\"EGFR\", \"CD9\", \"ITGA3\", \"ADAM12\", \"ADAM17\", \"MMP7\", \"APP\", \"AXL\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}