{"gene":"DLL4","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2007,"finding":"DLL4-Notch1 signaling between endothelial cells restricts tip-cell formation in response to VEGF, establishing the ratio between tip and stalk cells required for correct sprouting and branching. Genetic inactivation of one Dll4 allele or endothelial-specific Notch1 deletion increases tip cell numbers, while Notch activation reduces them, demonstrating lateral inhibition within angiogenic sprouts.","method":"Genetic mouse models (Dll4 haploinsufficiency, endothelial-specific Notch1 deletion), gamma-secretase inhibitors, soluble Jagged1 peptide treatment, retinal vascular imaging","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple genetic loss-of-function and pharmacological gain-of-function approaches in vivo, replicated across multiple labs","pmids":["17259973"],"is_preprint":false},{"year":2009,"finding":"Jagged1 antagonizes DLL4-Notch signaling in cells expressing Fringe family glycosyltransferases: upon Notch glycosylation by Fringe, DLL4-Notch signaling is enhanced while Jagged1 has weak signaling capacity and competes with DLL4, establishing that the equilibrium between two Notch ligands with opposing roles regulates angiogenesis.","method":"Genetic mouse models of Jagged1 and Dll4 manipulation, Fringe glycosyltransferase functional studies, in vivo retinal angiogenesis assays","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal genetic and biochemical approaches, published in high-impact journal, broadly replicated","pmids":["19524514"],"is_preprint":false},{"year":2006,"finding":"Neutralizing DLL4 with a selective antibody renders endothelial cells hyperproliferative and causes defective cell fate specification/differentiation both in vitro and in vivo, demonstrating that DLL4-mediated Notch signaling regulates endothelial cell proliferation and differentiation during active vascularization.","method":"DLL4-selective neutralizing antibody, in vitro endothelial cell assays, in vivo tumor xenograft models, histological analysis","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 / Strong — antibody-based loss-of-function with multiple in vitro and in vivo readouts, replicated across tumor models","pmids":["17183323"],"is_preprint":false},{"year":2017,"finding":"Genetic experiments in postnatal mice reveal that the level of active Notch signaling is more important than direct DLL4-mediated cell-cell communication. Endothelial tip cells retain their function without Dll4 and are not replaced by adjacent Dll4-positive cells. Instead, Dll4-Notch signaling directs tip-derived endothelial cells into developing arteries, coupling sprouting angiogenesis and artery formation.","method":"Conditional genetic targeting of endothelial tip cells in vivo, mosaic Dll4 deletion, postnatal retinal angiogenesis analysis","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with cell-type-specific in vivo models and multiple phenotypic readouts","pmids":["28714968"],"is_preprint":false},{"year":2006,"finding":"Hypoxia via HIF-1alpha induces DLL4 expression and downstream Notch target genes Hey1 and Hey2. The activated Dll4-Notch-Hey2 signaling cascade leads to repression of COUP-TFII in endothelial progenitor cells, promoting arterial cell fate decision. Hey factors provide negative feedback by repressing HIF-1alpha-induced gene expression.","method":"Promoter analysis, HIF-1alpha induction in cell lines, endothelial progenitor cell culture, gene expression analysis","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — promoter analysis combined with cell-based functional assays, single lab","pmids":["17045587"],"is_preprint":false},{"year":2010,"finding":"Beta-catenin (Wnt signaling) upregulates Dll4 transcription and strongly increases Notch signaling in endothelium, leading to functional and morphological vascular alterations including lack of vascular remodeling and loss of venous identity. Both in vivo and in vitro data establish a mechanistic link between Wnt and Notch signaling via DLL4.","method":"Endothelial-specific stabilization of beta-catenin in mice, in vitro endothelial cell assays, DLL4 promoter transcriptional analysis","journal":"Developmental cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo and in vitro convergent evidence, single lab","pmids":["20627076"],"is_preprint":false},{"year":2011,"finding":"DLL4-mediated Notch signaling in tumor endothelium increases large vessel formation, reduces VEGFR2 expression in large blood vessels, decreases VEGFR3 overall, and decreases hypoxia-induced VEGF while increasing VEGFR1 in tumor stroma — multiple mechanisms by which DLL4-Notch confers resistance to bevacizumab (anti-VEGF) therapy.","method":"Retroviral DLL4 transduction of glioblastoma cells, tumor xenografts, bevacizumab treatment, gamma-secretase inhibitor treatment, molecular analysis of resistance mechanisms","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo tumor model with multiple molecular mechanistic readouts, single lab","pmids":["21803743"],"is_preprint":false},{"year":2009,"finding":"DLL4 on dendritic cells is induced by pathogen-associated signals through TLR activation (but not by early inflammatory cytokines IL-1 and IL-18), and DLL4 signaling upregulates Rorc expression in T cells and directly targets Rorc and Il17 gene promoters to promote Th17 differentiation. DLL4 also inhibits Th2 cytokine production.","method":"In vitro T cell differentiation co-culture assays, siRNA knockdown, Notch signaling inhibition, gene promoter analysis","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — promoter analysis combined with functional differentiation assays, single lab","pmids":["19494260"],"is_preprint":false},{"year":2011,"finding":"DLL4-Notch1 signaling blockade disrupts postnatal lymphatic development by downregulating EphrinB2 expression (required for VEGFR3/VEGFC signaling), resulting in reduced lymphangiogenic sprouting, dilation of collecting lymphatic vessels with reduced mural cell coverage, and impaired wound healing/lymphangiogenesis.","method":"Function-blocking antibodies against Notch1 and Dll4 in mice, lymphatic development analysis, EphrinB2 expression analysis, wound healing model","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — antibody-based loss-of-function with multiple vascular phenotype readouts, single lab","pmids":["21700774"],"is_preprint":false},{"year":2011,"finding":"DLL4/Notch pathway regulates vessel regression by modulating vasoconstriction and blood flow: Dll4/Notch inhibition upregulates vasodilators (adrenomedullin) and suppresses vasoconstrictors (angiotensinogen), maintaining blood flow and preventing capillary regression. Angiotensin II induces rapid nonperfusion and regression of developing retinal capillaries.","method":"Genetic and pharmacologic Dll4/Notch inhibition in retinal oxygen-induced retinopathy model, Notch-regulated ankyrin repeat protein deletion, vasoactive molecule expression analysis","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and pharmacological approaches with mechanistic downstream pathway identification, single lab","pmids":["21498671"],"is_preprint":false},{"year":2015,"finding":"DLL4 expression in intestinal lacteals requires activation of VEGFR3 and VEGFR2, and genetic inactivation of Dll4 in lymphatic endothelial cells leads to lacteal regression and impaired dietary fat uptake, demonstrating a role for continuous DLL4 signaling in adult lymphatic vessel maintenance and function.","method":"Lymphatic endothelial cell-specific genetic inactivation of Dll4, VEGFR2/3 blocking experiments, dietary fat absorption assays, high-resolution intestinal stroma analysis","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Strong — tissue-specific genetic deletion with functional dietary fat absorption readout and upstream receptor pathway identification","pmids":["26529256"],"is_preprint":false},{"year":2011,"finding":"Adhesion of endothelial cells to laminin-111 via alpha2beta1 and alpha6beta1 integrins triggers DLL4 expression, leading to subsequent Notch pathway activation. Foxc2 transcription is required but not sufficient for DLL4 induction. VEGF stimulates laminin gamma1 deposition, which leads to integrin signaling and DLL4 induction. Loss of integrins alpha2 or alpha6 mimics DLL4 silencing effects.","method":"siRNA knockdown of integrins, endothelial cell adhesion assays on laminin-111, 3D matrigel sprouting assay, VEGF stimulation experiments","journal":"Circulation research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple siRNA knockdowns with functional sprouting assay readout, single lab","pmids":["21474814"],"is_preprint":false},{"year":2015,"finding":"Synaptojanin-2 binding protein (SYNJ2BP) physically interacts with the PDZ binding motif of DLL4 (and DLL1 but not Jagged-1), is preferentially expressed in stalk cells, enhances DLL4 protein stability, and promotes Notch signaling in endothelial cells. SYNJ2BP enables DLL4 interaction with Nectin-2 at adherens junctions.","method":"Co-immunoprecipitation, protein stability assays, siRNA knockdown, in vivo vascular density analysis in immunocompromised mice, in vitro endothelial cell assays","journal":"Circulation research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding demonstrated by Co-IP with functional stability and signaling consequences, single lab","pmids":["24025447"],"is_preprint":false},{"year":2014,"finding":"DLL4-containing exosomes can travel through 3D collagen matrix, transfer DLL4 protein to distant endothelial tip cells, activate Notch signaling in recipient cells, cause tip cell filopodia retraction, suppress sprout formation, increase endothelial cell motility, and suppress endothelial cell proliferation.","method":"3D microfluidic device, time-lapse confocal microscopy, exosome isolation and application, Notch signaling readouts","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct live imaging of exosome transfer with functional and signaling readouts, single lab","pmids":["24504253"],"is_preprint":false},{"year":2010,"finding":"At limiting expression levels, DLL4 maintains the ability to inhibit B lineage choice and induce T lineage commitment at lower expression levels than DLL1. DLL4 expressed at physiological levels supports T lineage cells and is permissive for myeloid cells while still inhibiting B lymphopoiesis. These properties correlate with DLL4's more efficient induction of Notch target genes and inhibition of B/myeloid-specific transcription factors.","method":"OP9 stromal cell lines expressing incrementally discrete levels of Dll1 or Dll4, hematopoietic progenitor coculture, Notch target gene expression analysis, lineage output analysis","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — quantitative dose-response comparison between DLL1 and DLL4 with functional lineage commitment readout, single lab","pmids":["20548034"],"is_preprint":false},{"year":2015,"finding":"DLL4/Notch1 and BMP9/ALK1 signaling pathways are interdependent: canonical BMP9 signaling via ALK1-Smad1/5/9 is disrupted by inhibition of Notch signaling, and DLL4 activity is suppressed when the ALK1-Smad1/5/9 pathway is inhibited. BMP9/DLL4 synergistically induces complete endothelial quiescence requiring P27KIP1 and upregulation of thrombospondin-1.","method":"Human endothelial cell stimulation with BMP9 and DLL4, Notch and ALK1 inhibitors, Dll4+/- mouse vascular analysis, proteomics","journal":"Arteriosclerosis, thrombosis, and vascular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal pathway inhibition experiments with proteomics, in vitro and in vivo convergent data, single lab","pmids":["26471266"],"is_preprint":false},{"year":2016,"finding":"DLL4 expression fluctuates in individual endothelial cells within sprouting vessels. High VEGF or DLL4 overexpression leads to Notch-dependent synchronization of DLL4 fluctuations within clusters, switching vessels from branching to expansion. Normal asynchronous Dll4 oscillations drive heterogeneity and branching, while synchronization drives vessel expansion.","method":"Live imaging in mouse retina in vivo, mouse embryonic stem cell-derived sprouting assays, DLL4 overexpression, Notch inhibition","journal":"eLife","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo and in vitro live imaging with genetic/pharmacological perturbations, single lab","pmids":["27074663"],"is_preprint":false},{"year":2015,"finding":"Leader cell identity during collective cell migration is dynamically regulated by DLL4 signaling through Notch1 and cellular stress. DLL4 is induced in leader cells after creation of a cell-free region, and leader cells are regulated via Notch1-DLL4 lateral inhibition. Mechanical stress inhibits DLL4 expression and leader cell formation.","method":"Single-cell gene expression analysis, computational modeling, time-lapse microscopy, Notch1-DLL4 inhibition","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — single-cell analysis combined with live imaging and perturbation experiments, single lab","pmids":["25766473"],"is_preprint":false},{"year":2015,"finding":"CCM1 silencing in endothelial cells causes decreased Notch3 activity in cocultured pericytes. Endothelial DLL4 stimulates Notch3 receptors on human brain pericytes; active Notch3 induces expression of PDGFRB2, N-Cadherin, HBEGF, TGFB1, NG2, and S1P genes, enhances pericyte adhesion to endothelial cells, limits pericyte migration/invasion, and enhances pericyte antiangiogenic function.","method":"Genetic manipulation of primary human endothelial cells and brain pericytes, Ccm1/Ccm2 endothelial-specific ablation in mouse models, Notch3 siRNA knockdown, pericyte-endothelial coculture","journal":"Stroke","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell-type-specific genetic manipulation with functional coculture readouts and in vivo CCM mouse models, single lab","pmids":["25791711"],"is_preprint":false},{"year":2018,"finding":"Multiple PDZ domain protein (MPDZ) physically interacts with the intracellular carboxyterminus of DLL4 (and DLL1) and enables their interaction with the adherens junction protein Nectin-2. MPDZ inactivation leads to impaired Notch signaling activity and increased blood vessel sprouting; tumor angiogenesis was enhanced upon endothelial-specific MPDZ inactivation.","method":"Co-immunoprecipitation, MPDZ gene inactivation in cellular models, embryonic mouse hindbrain vascular analysis, tumor angiogenesis models","journal":"eLife","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding demonstrated plus in vivo phenotype, single lab","pmids":["29620522"],"is_preprint":false},{"year":2019,"finding":"LPA4/LPA6-mediated Gα12/Gα13-Rho-ROCK signaling activates YAP/TAZ in endothelial cells; YAP/TAZ knockdown increases β-catenin- and NICD-mediated endothelial DLL4 expression. LPA4/LPA6 or YAP/TAZ knockdown blocks EC sprouting, rescued by Notch inhibitor, demonstrating that LPA-YAP/TAZ signaling promotes angiogenesis by repressing DLL4.","method":"Endothelial-specific Lpa4;Lpa6 double knockout mice, siRNA knockdown of YAP/TAZ, fibrin gel sprouting assay, retinal angiogenesis analysis, Notch inhibition rescue experiments","journal":"The Journal of clinical investigation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic double knockout with multiple in vitro and in vivo validation experiments, single lab","pmids":["31335323"],"is_preprint":false},{"year":2020,"finding":"Slug (SNAI2) transcription factor suppresses DLL4-Notch signaling in angiogenic endothelial cells, thereby promoting VEGFR2 expression. EC-specific Slug re-expression or reduced Notch signaling (gamma-secretase inhibition or Dll4 loss) rescues retinal angiogenesis in Slug knockout mice. Endothelial Slug is activated by SDF1alpha via CXCR4 and MAP kinase ERK5.","method":"Slug knockout mice, EC-specific re-expression, gamma-secretase inhibition, Dll4 loss-of-function, VEGF signaling inhibition, CXCR4/ERK5 pathway analysis","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic rescue experiments with epistasis analysis and upstream pathway identification, single lab","pmids":["33106502"],"is_preprint":false},{"year":2019,"finding":"Indoxyl sulfate induces DLL4 protein expression in macrophages via inhibition of the ubiquitin-proteasome pathway through the deubiquitinating enzyme USP5, triggering Notch signaling. Macrophage uptake of indoxyl sulfate is mediated by OATP2B1; DLL4 antibody and OATP2B1 siRNA inhibit proinflammatory macrophage activation and atherosclerotic lesion development in mice.","method":"In vitro macrophage treatment with indoxyl sulfate, global proteomics, siRNA knockdown (macrophage-targeted lipid nanoparticles), 5/6 nephrectomy mouse model, Dll4 antibody treatment, Ldlr-/- atherosclerosis model","journal":"Circulation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — proteomics-guided mechanistic validation with in vivo macrophage-targeted siRNA and antibody approaches, single lab","pmids":["30586693"],"is_preprint":false},{"year":2019,"finding":"In the context of diabetic wounds, high glucose levels activate a positive Dll4-Notch1 feedback loop, and Notch1 inactivation specifically in keratinocytes cancels the repressive effects of this loop on wound healing in diabetes, demonstrating that the Dll4-Notch1 loop in keratinocytes impairs diabetic wound healing.","method":"Genetic loss-of-function in diabetic mouse models, keratinocyte-specific Notch1 inactivation, local Notch signaling inhibition, wound healing analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell-type-specific genetic loss-of-function with functional wound healing readout, single lab","pmids":["30886104"],"is_preprint":false},{"year":2020,"finding":"Both antibody-based and genetic inactivation of Dll4 or Notch1 induces hyperpermeability in arterial endothelial cells by increasing transcytosis without junctional destabilization. Endothelial Sox17 deletion represses Dll4 in retinal arteries, phenocopying Dll4 blockade. Dll4 blocking activates SREBP1-mediated lipogenic transcription and caveolae formation. Inhibition of SREBP1 or VEGF-VEGFR2 attenuates Dll4 blockade-driven retinal leakage, establishing a Sox17-Dll4-SREBP1 axis controlling transcytosis independently of tight junctions.","method":"Antibody-based and genetic Dll4/Notch1 inactivation, Sox17 endothelial deletion, transcriptomic profiling, SREBP1 inhibition, VEGFR2 inhibition, hypertension-induced retinal edema model","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal genetic and pharmacological approaches identifying a novel transcytosis mechanism with pathway validation, single rigorous study","pmids":["32078435"],"is_preprint":false},{"year":2022,"finding":"In atrophic conditions, microvascular endothelium upregulates and releases the Notch ligand DLL4, which activates muscular Notch2 without direct cell-cell contact. Inhibition of the Dll4-Notch2 axis substantially prevents disuse- or diabetes-induced muscle atrophy and promotes overloading-induced muscle hypertrophy in mice, establishing an endothelial-to-muscle signaling axis controlling skeletal muscle mass.","method":"Myofiber Notch2 conditional knockout, Dll4 inhibition, disuse and diabetic atrophy mouse models, mechanical overload model, DLL4/Notch2 axis analysis","journal":"Nature metabolism","confidence":"High","confidence_rationale":"Tier 2 / Strong — tissue-specific genetic loss-of-function with multiple in vivo functional readouts identifying a novel inter-cellular signaling mechanism","pmids":["35228746"],"is_preprint":false},{"year":2015,"finding":"DLL4 is expressed in a sub-population of bipotent hematoendothelial progenitors (HEPs) in hESCs and segregates their hematopoietic versus endothelial potential. DLL4-high HEPs are enriched for endothelial potential, while DLL4-low/-negative HEPs are committed to hematopoietic lineage. DLL4 stimulation enhances hematopoietic differentiation of HEPs and increases clonogenic hematopoietic progenitors.","method":"hESC differentiation, clonal analysis, transcriptome analysis, confocal microscopy of embryoid bodies, DLL4 stimulation of HEPs","journal":"Leukemia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clonal and transcriptome analysis with functional stimulation experiments, single lab","pmids":["25778099"],"is_preprint":false},{"year":2020,"finding":"Dll4 acts as a negative regulator of intra-aortic hematopoietic cluster (IAHC) recruitment: blocking Dll4 promotes entrance of new hemogenic Gfi1+ cells into IAHC and increases the number of cells acquiring HSC activity. IACHs form through a two-step process where Dll4 inhibits the recruitment phase.","method":"Live imaging of organotypic slice cultures, clonal analysis, mathematical modeling, Dll4 blocking experiments in mouse embryos","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live imaging combined with clonal analysis and mathematical modeling, single lab","pmids":["32149421"],"is_preprint":false},{"year":2018,"finding":"DLL4 preferentially activates NOTCH1 over NOTCH2, whereas DLL1 is equally effective in activating NOTCH1 and NOTCH2. The discriminating potential lies in the region between the N-terminus and EGF repeat three of the ligand ectodomain. The ectodomains dictate selective ligand function in vivo during somitogenesis.","method":"Cellular co-culture signaling assays, biochemical binding studies, chimeric ligand knock-in mouse models, somitogenesis and myogenesis analysis","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — biochemical binding studies, chimeric ligand functional assays, and in vivo genetic validation in multiple contexts","pmids":["30289388"],"is_preprint":false},{"year":2015,"finding":"DLL4, but not DLL1, is an efficient cis-inhibitor of Notch signaling, causing reduced net activation of Notch in cells co-expressing ligand and receptor. This differential cis-inhibitory property contributes to context-dependent functional divergence between DLL1 and DLL4.","method":"Conditional overexpression from same genomic locus (Hprt), Dll1Dll4 knock-in mice, in vitro Notch signaling assays for cis-inhibition and trans-activation","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro signaling assays combined with in vivo genetic models, single lab","pmids":["26114479"],"is_preprint":false},{"year":2018,"finding":"LPS/TLR4 signaling induces ERK phosphorylation, which causes FOXC2-ERK protein ligation and ERK-dependent FOXC2 serine/threonine phosphorylation, subsequently activating DLL4 gene expression. FOXC2 binds to the DLL4 promoter in vivo. ERK inhibition or FOXC2 siRNA attenuates LPS-induced DLL4 expression and angiogenic sprouting.","method":"TLR4/LPS stimulation in human lung endothelial cells, ERK inhibition, ERK-2 dominant negative transfection, FOXC2-siRNA, in vivo mouse lung analysis, FOXC2+/- mice","journal":"The Journal of physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — promoter binding combined with multiple loss-of-function approaches in vitro and in vivo, single lab","pmids":["29380370"],"is_preprint":false},{"year":2019,"finding":"The DLL4 5'-UTR harbors an Internal Ribosomal Entry Site (IRES) that is efficiently utilized during hypoxia and ER stress (cap-independent translation). PERK kinase (activated by ER stress) drives DLL4 IRES-mediated translation, and hnRNP-A1 acts as an IRES-Trans-Acting Factor (ITAF) participating in IRES-dependent DLL4 translation during ER stress.","method":"IRES reporter assays, PERK inhibition/activation, hnRNP-A1 characterization, hypoxia and ER stress treatment of cells","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reporter assays for IRES activity combined with identification of specific trans-acting factors, single lab","pmids":["30691003"],"is_preprint":false},{"year":2012,"finding":"KSHV vGPCR upregulates DLL4 through an ERK-dependent mechanism in lymphatic endothelial cells, while vFLIP induces JAG1 through NFkappaB-dependent signaling. Both ligands signal through NOTCH4 and suppress cell cycle genes in adjacent lymphatic endothelial cells to induce quiescence.","method":"KSHV gene expression in lymphatic endothelial cells, ERK and NFkappaB pathway inhibition, gene expression profiling, functional Notch signaling assays","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — specific viral gene attribution with pathway inhibitor validation and gene expression profiling, single lab","pmids":["19816565"],"is_preprint":false},{"year":2019,"finding":"TMZ treatment promotes nuclear translocation of MMP14 followed by extracellular release of DLL4. Released DLL4 stimulates cleavage of Notch3, its nuclear translocation, and induction of sphering capacity and stemness in glioblastoma cells.","method":"Multiple PDX GBM models and glioma cell lines, MMP14 expression/localization analysis, Kiloplex ELISA-based protein array, DLL4/Notch3 functional studies","journal":"International journal of cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic identification of MMP14-DLL4-Notch3 axis with multiple models, single lab","pmids":["31443114"],"is_preprint":false},{"year":2019,"finding":"Soluble DLL4 activates Notch signaling in endothelial cells, increases VE-cadherin expression at intercellular junctions (but not ZO-1), and decreases vascular permeability. This permeability reduction acts through a cAMP/PKA pathway: PKA inhibition reverses the DLL4-mediated permeability reduction and reduces Hey1 expression. PKA knockdown reduces VE-cadherin junctional expression.","method":"Recombinant soluble DLL4 treatment of EC monolayers, gamma-secretase inhibitor, PKA inhibitors, hydraulic conductivity in rat mesenteric microvessels in vivo, FITC-albumin permeability assays","journal":"American journal of physiology. Heart and circulatory physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro and in vivo vascular permeability assays with pharmacological pathway dissection, single lab","pmids":["30681366"],"is_preprint":false},{"year":2021,"finding":"In zebrafish valvulogenesis, blood flow activates Notch signaling, which drives lateral inhibition between endocardial cells mediated by DLL4. DLL4-positive endocardial cells ingress into the cardiac jelly to form an abluminal cell population in response to Wnt9a (produced via Erk5-Klf2-Wnt9a cascade). Mechanical stimulation activates parallel mechanosensitive signaling pathways (Notch and Klf2) that intersect to drive valve formation.","method":"Zebrafish genetic models, live imaging, DLL4-positive cell lineage tracking, Notch and Wnt9a pathway perturbation experiments","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis in zebrafish with live imaging validation, single lab","pmids":["34610316"],"is_preprint":false},{"year":2021,"finding":"Pre-existing embryonic coronary plexus expresses DLL4, and DLL4-NOTCH1 signaling mediates angiogenic expansion of this plexus to vascularize the expanding myocardium in neonates, and also revascularizes the regenerating neonatal heart. Ventricular endocardial cells do not contribute to new coronary vessels.","method":"Lineage-tracing, gain- and loss-of-function genetic experiments in mice, neonatal heart regeneration model","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — convergent lineage tracing and genetic gain/loss-of-function establishing DLL4-NOTCH1 as the mechanistic driver, single rigorous study","pmids":["34497373"],"is_preprint":false},{"year":2019,"finding":"Coronary arterial development requires a DLL4-Jag1-EphrinB2 signaling cascade: endocardial Jag1 removal blocks sinus venosus capillary sprouting, while Dll4 inactivation stimulates excessive capillary growth. Forced Dll4 expression or Mfng (glycosyltransferase) blocks coronary plexus remodeling and arterial differentiation. EphrinB2 is a critical effector of antagonistic Dll4 and Jag1 functions in arterial morphogenesis.","method":"Endocardial Jag1 conditional deletion, Dll4 inactivation, forced Dll4/Mfng expression, Efnb2 endocardial deletion, angiogenic rescue experiments in ventricular explants and human endothelial cells","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple genetic loss- and gain-of-function with mechanistic rescue experiments in vivo and in vitro, single rigorous study","pmids":["31789590"],"is_preprint":false},{"year":2018,"finding":"LDB2 (LIM-domain binding protein 2) regulates basal and VEGF-induced DLL4 expression in endothelial cells by binding to the DLL4 promoter region through oligomeric complexes with LMO2/TAL1/GATA2. LDB2 overexpression increases DLL4 expression; LDB2 knockdown decreases DLL4 expression and enhances endothelial sprouting.","method":"siRNA knockdown and overexpression of LDB2, DLL4 promoter binding assays, zebrafish ldb2-morpholino, in vitro sprouting/tubular network assays","journal":"BMB reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — promoter binding assays combined with loss- and gain-of-function and in vivo zebrafish model, single lab","pmids":["28946938"],"is_preprint":false},{"year":2022,"finding":"Affinity-matured DLL4 variant (DeltaMAX) binds human and murine Notch receptors with 500- to 1000-fold increased affinity compared to wild-type human DLL4. DeltaMAX potently activates Notch in plate-bound, bead-bound, and cellular formats. As a soluble decoy, DeltaMAX inhibits Notch in reporter and neuronal differentiation assays, demonstrating dual agonist/antagonist utility.","method":"Protein engineering/affinity maturation, in vitro Notch activation assays (plate-bound, bead-bound, cellular), reporter assays, neuronal differentiation assays, T cell stimulation assays","journal":"Nature chemical biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with quantitative binding measurements and multiple functional validation assays","pmids":["36050494"],"is_preprint":false},{"year":2023,"finding":"Epsin1 modulates the sorting of DLL4 into exosomes from tubular epithelial cells under high glucose conditions. Exosomes enriched with DLL4 are captured by macrophages and promote M1 macrophage activation via Notch1 (N1ICD) activation. Epsin1 knockdown reduces DLL4 in TEC-exosomes and inhibits macrophage N1ICD activation and iNOS expression.","method":"Mass spectrometry of urine exosomes, Epsin1 siRNA knockdown, in vitro THP-1 macrophage treatment with exosomes, in vivo C57BL/6 mouse model, western blot, db/db diabetic mice","journal":"Molecular therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic identification of Epsin1-mediated DLL4 exosomal sorting with in vitro and in vivo validation, single lab","pmids":["37016580"],"is_preprint":false},{"year":2012,"finding":"MEDI0639 (anti-DLL4 antibody) inhibits the binding of Notch1 to DLL4 via a novel epitope not previously described, reversing Notch1-mediated suppression of HUVEC growth in vitro. MEDI0639 promotes tubule formation in 3D endothelial outgrowth assay (disrupting Dll4-Notch axis), but inhibits tubule formation in 2D endothelial-fibroblast coculture. In vivo, MEDI0639 promotes human vessel formation and reduces mural cell coverage.","method":"Notch1-DLL4 binding inhibition assays, HUVEC growth assays, 3D and 2D angiogenesis assays, in vivo human endothelial cell angiogenesis assay in mice","journal":"Molecular cancer therapeutics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding inhibition combined with multiple in vitro and in vivo functional assays, single lab","pmids":["22679110"],"is_preprint":false},{"year":2024,"finding":"Palmitic acid induces macrophage DLL4 signaling, which in turn triggers senescence in vascular smooth muscle cells, reducing collagen synthesis and deposition, thus promoting atherosclerotic plaque instability. Macrophage-specific DLL4 knockout in atherosclerotic mice leads to reduced plaque burden and improved stability.","method":"Human cohort studies, macrophage-specific DLL4 knockout in atherosclerotic mouse models, palmitic acid treatment of macrophages, vascular smooth muscle cell senescence assays","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — macrophage-specific genetic knockout with mechanistic cellular downstream identification, single lab","pmids":["38346959"],"is_preprint":false},{"year":2012,"finding":"Dll4-Notch signaling in DLL4-expressing cancer cells (SCLC) plays a critical role in liver metastasis by regulating NF-κB signaling. Dll4-Fc (soluble DLL4) acts as a blocker, and Dll4-Fc-expressing cancer cells show downregulated NF-κB activities (both classical and alternative pathways) by reducing Notch1 signaling, resulting in reduced liver metastasis.","method":"Soluble Dll4-Fc generation, SCLC cell line transduction, mouse liver metastasis model, PCR array analysis, electrophoretic mobility shift assay for NF-κB activity, Notch1 signaling analysis","journal":"Molecular cancer therapeutics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional metastasis model combined with NF-κB pathway analysis, single lab","pmids":["22989420"],"is_preprint":false},{"year":2012,"finding":"Dll4 blockade (pharmacological or genetic) induces accumulation of thymic dendritic cells and CD4+CD25+FoxP3+ regulatory T cells in the thymic cortex. Dll4 blockade converts DN1 T cell progenitors to immature DCs that induce Treg differentiation through a Flt3-independent, DC-dependent mechanism requiring MHC II expression. This mechanism depends on transcriptional upregulation of PU.1, Irf-4, Irf-8, and CSF-1.","method":"Pharmacological Dll4 blockade, genetic inactivation models, thymectomy experiments, DC-T cell coculture, anti-Dll4 antibody treatment in type 1 diabetes model","journal":"The Journal of experimental medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and pharmacological loss-of-function with epistasis analysis, single lab","pmids":["22547652"],"is_preprint":false},{"year":2019,"finding":"VEGF165 inhibits pro-fibrotic differentiation of endometrial stromal cells via the DLL4/Notch4/Smad7 pathway; inhibiting Smad7 or Notch4 blocks the anti-fibrotic effect of VEGF165, demonstrating that DLL4 and Notch4 are essential downstream molecules for VEGF165's anti-fibrotic function.","method":"VEGF165 treatment of human primary endometrial stromal cells, conditional VEGF reduction in mice, Smad7 and Notch4 inhibition, TGFbeta1-induced fibrosis model","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple inhibitor experiments establishing epistatic relationship, single lab","pmids":["31515487"],"is_preprint":false}],"current_model":"DLL4 is a transmembrane Notch ligand expressed predominantly in arterial and tip-cell endothelium that signals in trans through Notch1 (and Notch2/3/4 in specific contexts) to laterally inhibit tip-cell formation and restrict angiogenic sprouting in response to VEGF; its expression is regulated by VEGF, HIF-1alpha, Wnt/beta-catenin, integrin-laminin, Fringe-mediated glycosylation (which modulates its competitive balance with Jagged1), and transcriptional regulators including FOXC2, LDB2, and Slug, while its protein levels are stabilized by SYNJ2BP and MPDZ at adherens junctions; downstream, DLL4-Notch signaling controls arterio-venous specification, lymphatic maintenance, transcytosis-dependent vascular barrier function (via Sox17-SREBP1), endothelial quiescence (in cooperation with BMP9/ALK1 and via P27KIP1), and skeletal muscle mass (through an endothelial-to-myofiber Dll4-Notch2 axis), as well as T-cell lineage commitment in the thymus and hematopoietic progenitor fate decisions."},"narrative":{"mechanistic_narrative":"DLL4 is a transmembrane Notch ligand that operates as the central rheostat of sprouting angiogenesis, signaling in trans through Notch1 to enforce lateral inhibition that fixes the ratio of tip to stalk cells during VEGF-driven vascular outgrowth [PMID:17259973, PMID:17183323]. Within this circuit DLL4 expression fluctuates in individual endothelial cells, and the synchronization versus asynchrony of these oscillations toggles vessels between expansion and branching [PMID:27074663]; the integrated level of Notch activity, more than direct cell-cell contact, determines outcome and channels tip-derived cells into arteries [PMID:28714968]. DLL4 selectively activates NOTCH1 over NOTCH2 through its N-terminal ectodomain and, unlike DLL1, acts as an efficient cis-inhibitor, properties that account for its distinct functional output [PMID:30289388, PMID:26114479]. Its signaling strength is set by a competitive balance with Jagged1 that is tuned by Fringe glycosyltransferases [PMID:19524514, PMID:31789590]. DLL4 transcription is driven by a convergent set of inputs—VEGF/HIF-1alpha-arterial programs [PMID:17045587], Wnt/beta-catenin [PMID:20627076], integrin-laminin adhesion acting through FOXC2 [PMID:21474814, PMID:29380370], and LDB2/LMO2/TAL1/GATA2 complexes [PMID:28946938]—and is repressed by Slug and by YAP/TAZ [PMID:33106502, PMID:31335323]; protein levels are further controlled post-transcriptionally by IRES-mediated translation under hypoxia/ER stress [PMID:30691003], by deubiquitination via USP5 [PMID:30586693], and by junctional stabilization through the PDZ proteins SYNJ2BP and MPDZ that link DLL4 to Nectin-2 at adherens junctions [PMID:24025447, PMID:29620522]. Through these mechanisms DLL4-Notch controls arterio-venous and coronary vascular morphogenesis [PMID:34497373, PMID:31789590], lymphatic sprouting and lacteal maintenance via EphrinB2 and VEGFR3 [PMID:21700774, PMID:26529256], endothelial quiescence in cooperation with BMP9/ALK1 and P27KIP1 [PMID:26471266], and vascular barrier function through a Sox17-Dll4-SREBP1 axis that limits transcytosis [PMID:32078435]. Beyond the vasculature DLL4 governs T-cell lineage commitment and thymic Treg generation [PMID:20548034, PMID:22547652], hematoendothelial fate decisions [PMID:25778099, PMID:32149421], and an endothelial-to-myofiber Dll4-Notch2 axis controlling skeletal muscle mass [PMID:35228746]; it is also exploited in tumor angiogenesis and metastasis and in macrophage-driven atherosclerosis [PMID:17183323, PMID:21803743, PMID:38346959].","teleology":[{"year":2006,"claim":"Establishing whether DLL4 is required for endothelial differentiation versus proliferation defined its core role as a brake on uncontrolled vascular growth.","evidence":"DLL4-selective neutralizing antibody with in vitro endothelial assays and tumor xenografts","pmids":["17183323"],"confidence":"High","gaps":["Did not resolve which Notch receptor mediates the effect","Mechanism of fate specification downstream of Notch not defined"]},{"year":2007,"claim":"Resolving how endothelial sprouts select tip versus stalk identity showed DLL4-Notch1 lateral inhibition sets the tip:stalk ratio in response to VEGF.","evidence":"Dll4 haploinsufficiency, endothelial Notch1 deletion, gamma-secretase inhibitors and retinal imaging in mice","pmids":["17259973"],"confidence":"High","gaps":["Did not explain how DLL4 levels are dynamically encoded across the sprout","Transcriptional control of DLL4 by VEGF left unaddressed"]},{"year":2009,"claim":"Defining why two Notch ligands have opposing angiogenic roles revealed that Fringe glycosylation tilts a DLL4-versus-Jagged1 competitive equilibrium.","evidence":"Genetic Jagged1/Dll4 manipulation and Fringe glycosyltransferase studies in mouse retinal angiogenesis","pmids":["19524514"],"confidence":"High","gaps":["Quantitative thresholds of ligand balance not defined","Structural basis of Fringe-dependent discrimination unresolved"]},{"year":2010,"claim":"Identifying upstream transcriptional drivers connected angiogenic signaling and Wnt to DLL4, and quantified DLL4's distinct potency in lineage commitment.","evidence":"Endothelial beta-catenin stabilization in mice plus DLL4 promoter analysis; OP9 dose-titration cocultures comparing DLL1 and DLL4","pmids":["20627076","20548034"],"confidence":"Medium","gaps":["Direct beta-catenin promoter occupancy not fully resolved","Molecular basis of DLL4's greater potency versus DLL1 not yet mapped"]},{"year":2011,"claim":"Mapping additional inducers and downstream effectors extended DLL4 control to integrin-laminin adhesion, lymphatic development, and vessel regression.","evidence":"Integrin siRNA with laminin adhesion and 3D sprouting assays; anti-Dll4/Notch1 antibodies in lymphatic and oxygen-induced retinopathy models","pmids":["21474814","21700774","21498671"],"confidence":"Medium","gaps":["FOXC2 shown necessary but not sufficient for DLL4 induction","Link between EphrinB2 regulation and VEGFR3 signaling mechanistically incomplete"]},{"year":2013,"claim":"Discovering DLL4 PDZ-motif binding partners explained how DLL4 protein is stabilized and positioned at adherens junctions to sustain Notch output.","evidence":"Co-IP, protein stability assays and siRNA in endothelial cells (SYNJ2BP); later MPDZ Co-IP plus in vivo hindbrain and tumor angiogenesis","pmids":["24025447","29620522"],"confidence":"Medium","gaps":["Single-lab Co-IP without independent reciprocal validation for each partner","How junctional localization tunes trans-signaling strength not quantified"]},{"year":2015,"claim":"A series of studies expanded DLL4 into adult homeostasis—lacteal maintenance, endothelial quiescence, pericyte regulation, and hematoendothelial fate.","evidence":"Lymphatic-specific Dll4 deletion with fat-absorption assays; BMP9/ALK1 reciprocal inhibition with proteomics; Notch3 pericyte cocultures; hESC clonal differentiation","pmids":["26529256","26471266","25791711","25778099"],"confidence":"Medium","gaps":["Interdependence of DLL4 and BMP9/ALK1 mechanistically partial","Context-specificity of which Notch receptor DLL4 engages not uniformly resolved"]},{"year":2016,"claim":"Live imaging of DLL4 dynamics reframed angiogenic patterning as an oscillatory system in which synchronization switches branching to expansion.","evidence":"In vivo retinal live imaging and ES-cell sprouting assays with DLL4 overexpression and Notch inhibition","pmids":["27074663"],"confidence":"Medium","gaps":["Molecular clock generating DLL4 oscillations not identified","Single-lab observation"]},{"year":2017,"claim":"Testing whether direct DLL4 contact or net Notch level dominates showed integrated Notch activity directs tip-derived cells into arteries.","evidence":"Mosaic Dll4 deletion and tip-cell genetic targeting in postnatal mouse retina","pmids":["28714968"],"confidence":"High","gaps":["How tip cells retain function without Dll4 not fully explained","Compensating ligand sources not defined"]},{"year":2018,"claim":"Biochemical and genetic dissection of the ectodomain established the molecular basis of DLL4 receptor selectivity and additional transcriptional regulators.","evidence":"Chimeric ligand knock-in mice and binding studies (NOTCH1 vs NOTCH2 selectivity); LDB2 promoter complexes; LPS-FOXC2-ERK promoter axis","pmids":["30289388","28946938","29380370"],"confidence":"High","gaps":["Structural detail of the discriminating N-terminal region not solved","Combinatorial logic among transcriptional inputs unresolved"]},{"year":2020,"claim":"Identifying the Sox17-Dll4-SREBP1 axis revealed a transcytosis-based, junction-independent mechanism by which DLL4 maintains the vascular barrier.","evidence":"Antibody and genetic Dll4/Notch1 inactivation, Sox17 deletion, transcriptomics and SREBP1 inhibition in retinal models","pmids":["32078435"],"confidence":"High","gaps":["Direct molecular link from NICD to SREBP1 activation not fully traced","Caveolae-formation step mechanistically incomplete"]},{"year":2022,"claim":"Demonstrating an endothelial-to-myofiber Dll4-Notch2 axis and engineering high-affinity DLL4 variants extended DLL4 biology beyond the vasculature and into reagent design.","evidence":"Myofiber Notch2 conditional knockout with atrophy/overload models; affinity-matured DeltaMAX variant with quantitative binding and functional assays","pmids":["35228746","36050494"],"confidence":"High","gaps":["How released DLL4 acts without cell-cell contact mechanistically open","In vivo therapeutic translation of engineered variants not established"]},{"year":2024,"claim":"Disease-context studies positioned DLL4 in macrophage-driven atherosclerosis and exosome-mediated intercellular Notch signaling.","evidence":"Macrophage-specific DLL4 knockout in atherosclerotic mice; Epsin1-dependent DLL4 exosome sorting with macrophage activation assays","pmids":["38346959","37016580"],"confidence":"Medium","gaps":["Mechanism by which DLL4 triggers VSMC senescence partial","Single-lab disease-context findings"]},{"year":null,"claim":"The structural basis and quantitative thresholds that convert DLL4 oscillation, cis-inhibition, and ligand competition into specific cell-fate outputs across tissues remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model linking DLL4 expression dynamics to receptor selection in vivo","Mechanism of contact-independent (soluble/exosomal) DLL4 signaling incompletely defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[0,2,28,39]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,1,28,29]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[1,29]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,28]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[13,40]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[33,34,43]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,28]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[3,36,37]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[7,14,44]}],"complexes":[],"partners":["NOTCH1","NOTCH2","NOTCH3","NOTCH4","SYNJ2BP","MPDZ","JAG1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NR61","full_name":"Delta-like protein 4","aliases":["Drosophila Delta homolog 4","Delta4"],"length_aa":685,"mass_kda":74.6,"function":"Involved in the Notch signaling pathway as Notch ligand (PubMed:11134954). Activates NOTCH1 and NOTCH4. Involved in angiogenesis; negatively regulates endothelial cell proliferation and migration and angiogenic sprouting (PubMed:20616313). Essential for retinal progenitor proliferation. Required for suppressing rod fates in late retinal progenitors as well as for proper generation of other retinal cell types (By similarity). During spinal cord neurogenesis, inhibits V2a interneuron fate (PubMed:17728344)","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q9NR61/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DLL4","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/DLL4","total_profiled":1310},"omim":[{"mim_id":"621120","title":"DELTA-LIKE NONCANONICAL NOTCH LIGAND 2; DLK2","url":"https://www.omim.org/entry/621120"},{"mim_id":"618806","title":"T-CELL LYMPHOPENIA, INFANTILE, WITH OR WITHOUT NAIL DYSTROPHY, AUTOSOMAL DOMINANT; TLIND","url":"https://www.omim.org/entry/618806"},{"mim_id":"616589","title":"ADAMS-OLIVER SYNDROME 6; AOS6","url":"https://www.omim.org/entry/616589"},{"mim_id":"616419","title":"ADHESION G PROTEIN-COUPLED RECEPTOR L4; ADGRL4","url":"https://www.omim.org/entry/616419"},{"mim_id":"615893","title":"NEURALIZED E3 UBIQUITIN PROTEIN LIGASE 1B; NEURL1B","url":"https://www.omim.org/entry/615893"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/DLL4"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"Q9NR61","domains":[{"cath_id":"2.60.40.3510","chopping":"27-188","consensus_level":"high","plddt":92.7613,"start":27,"end":188},{"cath_id":"2.10.25.140","chopping":"192-221","consensus_level":"medium","plddt":95.7867,"start":192,"end":221},{"cath_id":"2.10.25.10","chopping":"367-403","consensus_level":"medium","plddt":80.1984,"start":367,"end":403}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NR61","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NR61-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NR61-F1-predicted_aligned_error_v6.png","plddt_mean":74.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DLL4","jax_strain_url":"https://www.jax.org/strain/search?query=DLL4"},"sequence":{"accession":"Q9NR61","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NR61.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NR61/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NR61"}},"corpus_meta":[{"pmid":"17259973","id":"PMC_17259973","title":"Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis.","date":"2007","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/17259973","citation_count":1375,"is_preprint":false},{"pmid":"19524514","id":"PMC_19524514","title":"The notch ligands Dll4 and Jagged1 have opposing effects on angiogenesis.","date":"2009","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/19524514","citation_count":900,"is_preprint":false},{"pmid":"17183323","id":"PMC_17183323","title":"Inhibition of Dll4 signalling inhibits tumour growth by deregulating angiogenesis.","date":"2006","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/17183323","citation_count":792,"is_preprint":false},{"pmid":"21238454","id":"PMC_21238454","title":"Dll1- and dll4-mediated notch signaling are required for homeostasis of intestinal stem cells.","date":"2011","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/21238454","citation_count":352,"is_preprint":false},{"pmid":"28714968","id":"PMC_28714968","title":"Dll4 and Notch signalling couples sprouting angiogenesis and artery formation.","date":"2017","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/28714968","citation_count":331,"is_preprint":false},{"pmid":"19664991","id":"PMC_19664991","title":"DLL4 blockade inhibits tumor growth and reduces tumor-initiating cell frequency.","date":"2009","source":"Cell stem cell","url":"https://pubmed.ncbi.nlm.nih.gov/19664991","citation_count":322,"is_preprint":false},{"pmid":"20627076","id":"PMC_20627076","title":"The Wnt/beta-catenin pathway modulates vascular remodeling and specification by upregulating Dll4/Notch signaling.","date":"2010","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/20627076","citation_count":265,"is_preprint":false},{"pmid":"21803743","id":"PMC_21803743","title":"DLL4-Notch signaling mediates tumor resistance to anti-VEGF therapy in vivo.","date":"2011","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/21803743","citation_count":191,"is_preprint":false},{"pmid":"17045587","id":"PMC_17045587","title":"Hypoxia-mediated activation of Dll4-Notch-Hey2 signaling in endothelial progenitor cells and adoption of arterial cell fate.","date":"2006","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/17045587","citation_count":179,"is_preprint":false},{"pmid":"30586693","id":"PMC_30586693","title":"Uremic Toxin Indoxyl Sulfate Promotes Proinflammatory Macrophage Activation Via the Interplay of OATP2B1 and Dll4-Notch Signaling.","date":"2019","source":"Circulation","url":"https://pubmed.ncbi.nlm.nih.gov/30586693","citation_count":176,"is_preprint":false},{"pmid":"19494260","id":"PMC_19494260","title":"Regulation of T cell activation by Notch ligand, DLL4, promotes IL-17 production and Rorc activation.","date":"2009","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/19494260","citation_count":154,"is_preprint":false},{"pmid":"26529256","id":"PMC_26529256","title":"DLL4 promotes continuous adult intestinal lacteal regeneration and dietary fat transport.","date":"2015","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/26529256","citation_count":145,"is_preprint":false},{"pmid":"20548034","id":"PMC_20548034","title":"Direct comparison of Dll1- and Dll4-mediated Notch activation levels shows differential lymphomyeloid lineage commitment outcomes.","date":"2010","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/20548034","citation_count":132,"is_preprint":false},{"pmid":"25767274","id":"PMC_25767274","title":"Endothelial Jagged1 antagonizes Dll4 regulation of endothelial branching and promotes vascular maturation downstream of Dll4/Notch1.","date":"2015","source":"Arteriosclerosis, thrombosis, and vascular biology","url":"https://pubmed.ncbi.nlm.nih.gov/25767274","citation_count":107,"is_preprint":false},{"pmid":"25766473","id":"PMC_25766473","title":"Notch1-Dll4 signalling and mechanical force regulate leader cell formation during collective cell migration.","date":"2015","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/25766473","citation_count":104,"is_preprint":false},{"pmid":"21498671","id":"PMC_21498671","title":"The Dll4/Notch pathway controls postangiogenic blood vessel remodeling and regression by modulating vasoconstriction and blood flow.","date":"2011","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/21498671","citation_count":99,"is_preprint":false},{"pmid":"31335323","id":"PMC_31335323","title":"Lysophosphatidic acid-induced YAP/TAZ activation promotes developmental angiogenesis by repressing Notch ligand Dll4.","date":"2019","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/31335323","citation_count":97,"is_preprint":false},{"pmid":"21193546","id":"PMC_21193546","title":"Anti-DLL4 inhibits growth and reduces tumor-initiating cell frequency in colorectal tumors with oncogenic KRAS mutations.","date":"2010","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/21193546","citation_count":96,"is_preprint":false},{"pmid":"21474814","id":"PMC_21474814","title":"Laminin-binding integrins induce Dll4 expression and Notch signaling in endothelial cells.","date":"2011","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/21474814","citation_count":95,"is_preprint":false},{"pmid":"19844231","id":"PMC_19844231","title":"Expression of delta-like ligand 4 (Dll4) and markers of hypoxia in colon cancer.","date":"2009","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/19844231","citation_count":95,"is_preprint":false},{"pmid":"27074663","id":"PMC_27074663","title":"Synchronization of endothelial Dll4-Notch dynamics switch blood vessels from branching to expansion.","date":"2016","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/27074663","citation_count":93,"is_preprint":false},{"pmid":"33106502","id":"PMC_33106502","title":"Slug regulates the Dll4-Notch-VEGFR2 axis to control endothelial cell activation and angiogenesis.","date":"2020","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/33106502","citation_count":87,"is_preprint":false},{"pmid":"15923152","id":"PMC_15923152","title":"Expression of Dll4 during mouse embryogenesis suggests multiple developmental roles.","date":"2005","source":"Gene expression patterns : GEP","url":"https://pubmed.ncbi.nlm.nih.gov/15923152","citation_count":85,"is_preprint":false},{"pmid":"24504253","id":"PMC_24504253","title":"Dll4-containing exosomes induce capillary sprout retraction in a 3D microenvironment.","date":"2014","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/24504253","citation_count":84,"is_preprint":false},{"pmid":"21700774","id":"PMC_21700774","title":"The Notch1-Dll4 signaling pathway regulates mouse postnatal lymphatic development.","date":"2011","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/21700774","citation_count":80,"is_preprint":false},{"pmid":"26299364","id":"PMC_26299364","title":"Heterozygous Loss-of-Function Mutations in DLL4 Cause Adams-Oliver Syndrome.","date":"2015","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26299364","citation_count":76,"is_preprint":false},{"pmid":"30886104","id":"PMC_30886104","title":"Triggering of a Dll4-Notch1 loop impairs wound healing in diabetes.","date":"2019","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/30886104","citation_count":76,"is_preprint":false},{"pmid":"21092311","id":"PMC_21092311","title":"Combination of Dll4/Notch and Ephrin-B2/EphB4 targeted therapy is highly effective in disrupting tumor angiogenesis.","date":"2010","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/21092311","citation_count":75,"is_preprint":false},{"pmid":"24114288","id":"PMC_24114288","title":"Dll4-Notch signaling in regulation of tumor angiogenesis.","date":"2013","source":"Journal of cancer research and clinical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/24114288","citation_count":71,"is_preprint":false},{"pmid":"25791711","id":"PMC_25791711","title":"Cerebral Cavernous Malformation-1 Protein Controls DLL4-Notch3 Signaling Between the Endothelium and Pericytes.","date":"2015","source":"Stroke","url":"https://pubmed.ncbi.nlm.nih.gov/25791711","citation_count":65,"is_preprint":false},{"pmid":"17822956","id":"PMC_17822956","title":"Anti-Dll4 therapy: can we block tumour growth by increasing angiogenesis?","date":"2007","source":"Trends in molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/17822956","citation_count":61,"is_preprint":false},{"pmid":"17158094","id":"PMC_17158094","title":"Expression of Dll4 and CCL25 in Foxn1-negative epithelial cells in the post-natal thymus.","date":"2006","source":"International immunology","url":"https://pubmed.ncbi.nlm.nih.gov/17158094","citation_count":59,"is_preprint":false},{"pmid":"26471266","id":"PMC_26471266","title":"DLL4/Notch1 and BMP9 Interdependent Signaling Induces Human Endothelial Cell Quiescence via P27KIP1 and Thrombospondin-1.","date":"2015","source":"Arteriosclerosis, thrombosis, and vascular biology","url":"https://pubmed.ncbi.nlm.nih.gov/26471266","citation_count":55,"is_preprint":false},{"pmid":"31233801","id":"PMC_31233801","title":"Delta-like ligand 4/DLL4 regulates the capillarization of liver sinusoidal endothelial cell and liver fibrogenesis.","date":"2019","source":"Biochimica et biophysica acta. Molecular cell research","url":"https://pubmed.ncbi.nlm.nih.gov/31233801","citation_count":54,"is_preprint":false},{"pmid":"22952347","id":"PMC_22952347","title":"Anti-DLL4 has broad spectrum activity in pancreatic cancer dependent on targeting DLL4-Notch signaling in both tumor and vasculature cells.","date":"2012","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/22952347","citation_count":54,"is_preprint":false},{"pmid":"32042099","id":"PMC_32042099","title":"Crenigacestat, a selective NOTCH1 inhibitor, reduces intrahepatic cholangiocarcinoma progression by blocking VEGFA/DLL4/MMP13 axis.","date":"2020","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/32042099","citation_count":53,"is_preprint":false},{"pmid":"23239744","id":"PMC_23239744","title":"Cross-talk between leukemic and endothelial cells promotes angiogenesis by VEGF activation of the Notch/Dll4 pathway.","date":"2012","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/23239744","citation_count":52,"is_preprint":false},{"pmid":"19816565","id":"PMC_19816565","title":"KSHV manipulates Notch signaling by DLL4 and JAG1 to alter cell cycle genes in lymphatic endothelia.","date":"2009","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/19816565","citation_count":51,"is_preprint":false},{"pmid":"23787764","id":"PMC_23787764","title":"Implications of Dll4-Notch signaling activation in primary glioblastoma multiforme.","date":"2013","source":"Neuro-oncology","url":"https://pubmed.ncbi.nlm.nih.gov/23787764","citation_count":51,"is_preprint":false},{"pmid":"30116629","id":"PMC_30116629","title":"The Role of Dll4/Notch Signaling in Normal and Pathological Ocular Angiogenesis: Dll4 Controls Blood Vessel Sprouting and Vessel Remodeling in Normal and Pathological Conditions.","date":"2018","source":"Journal of ophthalmology","url":"https://pubmed.ncbi.nlm.nih.gov/30116629","citation_count":50,"is_preprint":false},{"pmid":"30975104","id":"PMC_30975104","title":"FKBPL and its peptide derivatives inhibit endocrine therapy resistant cancer stem cells and breast cancer metastasis by downregulating DLL4 and Notch4.","date":"2019","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/30975104","citation_count":50,"is_preprint":false},{"pmid":"22679110","id":"PMC_22679110","title":"MEDI0639: a novel therapeutic antibody targeting Dll4 modulates endothelial cell function and angiogenesis in vivo.","date":"2012","source":"Molecular cancer therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/22679110","citation_count":48,"is_preprint":false},{"pmid":"32078435","id":"PMC_32078435","title":"Dll4 Suppresses Transcytosis for Arterial Blood-Retinal Barrier Homeostasis.","date":"2020","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/32078435","citation_count":47,"is_preprint":false},{"pmid":"25778099","id":"PMC_25778099","title":"The Notch ligand DLL4 specifically marks human hematoendothelial progenitors and regulates their hematopoietic fate.","date":"2015","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/25778099","citation_count":47,"is_preprint":false},{"pmid":"22989420","id":"PMC_22989420","title":"Dll4-Fc, an inhibitor of Dll4-notch signaling, suppresses liver metastasis of small cell lung cancer cells through the downregulation of the NF-κB activity.","date":"2012","source":"Molecular cancer therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/22989420","citation_count":47,"is_preprint":false},{"pmid":"32149421","id":"PMC_32149421","title":"Notch ligand Dll4 impairs cell recruitment to aortic clusters and limits blood stem cell generation.","date":"2020","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/32149421","citation_count":45,"is_preprint":false},{"pmid":"18577711","id":"PMC_18577711","title":"Dll4 activation of Notch signaling reduces tumor vascularity and inhibits tumor growth.","date":"2008","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/18577711","citation_count":45,"is_preprint":false},{"pmid":"22547652","id":"PMC_22547652","title":"Dll4-Notch signaling in Flt3-independent dendritic cell development and autoimmunity in mice.","date":"2012","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/22547652","citation_count":42,"is_preprint":false},{"pmid":"27009216","id":"PMC_27009216","title":"Dll4 Inhibition plus Aflibercept Markedly Reduces Ovarian Tumor Growth.","date":"2016","source":"Molecular cancer therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/27009216","citation_count":42,"is_preprint":false},{"pmid":"37016580","id":"PMC_37016580","title":"Epsin1-mediated exosomal sorting of Dll4 modulates the tubular-macrophage crosstalk in diabetic nephropathy.","date":"2023","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/37016580","citation_count":40,"is_preprint":false},{"pmid":"35228746","id":"PMC_35228746","title":"The endothelial Dll4-muscular Notch2 axis regulates skeletal muscle mass.","date":"2022","source":"Nature metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/35228746","citation_count":40,"is_preprint":false},{"pmid":"24025447","id":"PMC_24025447","title":"Synaptojanin-2 binding protein stabilizes the Notch ligands DLL1 and DLL4 and inhibits sprouting angiogenesis.","date":"2013","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/24025447","citation_count":39,"is_preprint":false},{"pmid":"23950980","id":"PMC_23950980","title":"Association of Dll4/notch and HIF-1a -VEGF signaling in the angiogenesis of missed abortion.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23950980","citation_count":38,"is_preprint":false},{"pmid":"26739060","id":"PMC_26739060","title":"MMGZ01, an anti-DLL4 monoclonal antibody, promotes nonfunctional vessels and inhibits breast tumor growth.","date":"2015","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/26739060","citation_count":37,"is_preprint":false},{"pmid":"38346959","id":"PMC_38346959","title":"Palmitic acid in type 2 diabetes mellitus promotes atherosclerotic plaque vulnerability via macrophage Dll4 signaling.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/38346959","citation_count":36,"is_preprint":false},{"pmid":"36001668","id":"PMC_36001668","title":"DLL4 and VCAM1 enhance the emergence of T cell-competent hematopoietic progenitors from human pluripotent stem cells.","date":"2022","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/36001668","citation_count":36,"is_preprint":false},{"pmid":"24949865","id":"PMC_24949865","title":"The expression of VEGF and Dll4/Notch pathway molecules in ovarian cancer.","date":"2014","source":"Clinica chimica acta; international journal of clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24949865","citation_count":35,"is_preprint":false},{"pmid":"25355291","id":"PMC_25355291","title":"DLL4 regulates NOTCH signaling and growth of T acute lymphoblastic leukemia cells in NOD/SCID mice.","date":"2014","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/25355291","citation_count":35,"is_preprint":false},{"pmid":"34610316","id":"PMC_34610316","title":"Mechanosensitive Notch-Dll4 and Klf2-Wnt9 signaling pathways intersect in guiding valvulogenesis in zebrafish.","date":"2021","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/34610316","citation_count":35,"is_preprint":false},{"pmid":"26114479","id":"PMC_26114479","title":"Context-Dependent Functional Divergence of the Notch Ligands DLL1 and DLL4 In Vivo.","date":"2015","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26114479","citation_count":35,"is_preprint":false},{"pmid":"29620522","id":"PMC_29620522","title":"MPDZ promotes DLL4-induced Notch signaling during angiogenesis.","date":"2018","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/29620522","citation_count":34,"is_preprint":false},{"pmid":"24931473","id":"PMC_24931473","title":"Endothelial Delta-like 4 (DLL4) promotes renal cell carcinoma hematogenous metastasis.","date":"2014","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/24931473","citation_count":31,"is_preprint":false},{"pmid":"34497373","id":"PMC_34497373","title":"Perinatal angiogenesis from pre-existing coronary vessels via DLL4-NOTCH1 signalling.","date":"2021","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/34497373","citation_count":31,"is_preprint":false},{"pmid":"30289388","id":"PMC_30289388","title":"The ectodomains determine ligand function in vivo and selectivity of DLL1 and DLL4 toward NOTCH1 and NOTCH2 in vitro.","date":"2018","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/30289388","citation_count":31,"is_preprint":false},{"pmid":"31616059","id":"PMC_31616059","title":"Specific NOTCH1 antibody targets DLL4-induced proliferation, migration, and angiogenesis in NOTCH1-mutated CLL cells.","date":"2019","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/31616059","citation_count":30,"is_preprint":false},{"pmid":"26808710","id":"PMC_26808710","title":"Arterialization and anomalous vein wall remodeling in varicose veins is associated with upregulated FoxC2-Dll4 pathway.","date":"2016","source":"Laboratory investigation; a journal of technical methods and pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26808710","citation_count":30,"is_preprint":false},{"pmid":"29380370","id":"PMC_29380370","title":"Endothelial immune activation programmes cell-fate decisions and angiogenesis by inducing angiogenesis regulator DLL4 through TLR4-ERK-FOXC2 signalling.","date":"2018","source":"The Journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/29380370","citation_count":30,"is_preprint":false},{"pmid":"31789590","id":"PMC_31789590","title":"Coronary arterial development is regulated by a Dll4-Jag1-EphrinB2 signaling cascade.","date":"2019","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/31789590","citation_count":29,"is_preprint":false},{"pmid":"24860699","id":"PMC_24860699","title":"Distinct expression patterns of Notch ligands, Dll1 and Dll4, in normal and inflamed mice intestine.","date":"2014","source":"PeerJ","url":"https://pubmed.ncbi.nlm.nih.gov/24860699","citation_count":29,"is_preprint":false},{"pmid":"30681366","id":"PMC_30681366","title":"Activation of Notch signaling by soluble Dll4 decreases vascular permeability via a cAMP/PKA-dependent pathway.","date":"2019","source":"American journal of physiology. Heart and circulatory physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30681366","citation_count":29,"is_preprint":false},{"pmid":"34543658","id":"PMC_34543658","title":"Substrate stiffness modulates endothelial cell function via the YAP-Dll4-Notch1 pathway.","date":"2021","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/34543658","citation_count":27,"is_preprint":false},{"pmid":"32029480","id":"PMC_32029480","title":"DLL1- and DLL4-Mediated Notch Signaling Is Essential for Adult Pancreatic Islet Homeostasis.","date":"2020","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/32029480","citation_count":26,"is_preprint":false},{"pmid":"32111499","id":"PMC_32111499","title":"Highly Expressed DLL4 and JAG1: Their Role in Incidence of Breast Cancer Metastasis.","date":"2020","source":"Archives of medical research","url":"https://pubmed.ncbi.nlm.nih.gov/32111499","citation_count":26,"is_preprint":false},{"pmid":"27301650","id":"PMC_27301650","title":"A humanized anti-DLL4 antibody promotes dysfunctional angiogenesis and inhibits breast tumor growth.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27301650","citation_count":26,"is_preprint":false},{"pmid":"23898884","id":"PMC_23898884","title":"Clinical implications of DLL4 expression in gastric cancer.","date":"2013","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/23898884","citation_count":25,"is_preprint":false},{"pmid":"26131280","id":"PMC_26131280","title":"HDAC5 promotes colorectal cancer cell proliferation by up-regulating DLL4 expression.","date":"2015","source":"International journal of clinical and experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/26131280","citation_count":25,"is_preprint":false},{"pmid":"38462037","id":"PMC_38462037","title":"LRP1 induces anti-PD-1 resistance by modulating the DLL4-NOTCH2-CCL2 axis and redirecting M2-like macrophage polarisation in bladder cancer.","date":"2024","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/38462037","citation_count":24,"is_preprint":false},{"pmid":"27639599","id":"PMC_27639599","title":"DLL4+ dendritic cells: Key regulators of Notch Signaling in effector T cell responses.","date":"2016","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/27639599","citation_count":24,"is_preprint":false},{"pmid":"25041739","id":"PMC_25041739","title":"Dengue virus up-regulates expression of notch ligands Dll1 and Dll4 through interferon-β signalling pathway.","date":"2015","source":"Immunology","url":"https://pubmed.ncbi.nlm.nih.gov/25041739","citation_count":24,"is_preprint":false},{"pmid":"37993804","id":"PMC_37993804","title":"Lactate receptor GPR81 drives breast cancer growth and invasiveness through regulation of ECM properties and Notch ligand DLL4.","date":"2023","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/37993804","citation_count":23,"is_preprint":false},{"pmid":"30130526","id":"PMC_30130526","title":"Propranolol inhibits proliferation and invasion of hemangioma-derived endothelial cells by suppressing the DLL4/Notch1/Akt pathway.","date":"2018","source":"Chemico-biological interactions","url":"https://pubmed.ncbi.nlm.nih.gov/30130526","citation_count":23,"is_preprint":false},{"pmid":"31443114","id":"PMC_31443114","title":"TMZ regulates GBM stemness via MMP14-DLL4-Notch3 pathway.","date":"2019","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/31443114","citation_count":22,"is_preprint":false},{"pmid":"26589434","id":"PMC_26589434","title":"Balancing Efficacy and Safety of an Anti-DLL4 Antibody through Pharmacokinetic Modulation.","date":"2015","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/26589434","citation_count":22,"is_preprint":false},{"pmid":"28262821","id":"PMC_28262821","title":"Notch Ligand DLL4 Alleviates Allergic Airway Inflammation via Induction of a Homeostatic Regulatory Pathway.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28262821","citation_count":22,"is_preprint":false},{"pmid":"32918765","id":"PMC_32918765","title":"TRIM28 regulates sprouting angiogenesis through VEGFR-DLL4-Notch signaling circuit.","date":"2020","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/32918765","citation_count":21,"is_preprint":false},{"pmid":"28288569","id":"PMC_28288569","title":"Endothelial Dll4 overexpression reduces vascular response and inhibits tumor growth and metastasization in vivo.","date":"2017","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/28288569","citation_count":21,"is_preprint":false},{"pmid":"21209419","id":"PMC_21209419","title":"Role of the DLL4-NOTCH system in PGF2alpha-induced luteolysis in the pregnant rat.","date":"2011","source":"Biology of reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/21209419","citation_count":21,"is_preprint":false},{"pmid":"31515487","id":"PMC_31515487","title":"Vascular endothelial growth factor 165 inhibits pro-fibrotic differentiation of stromal cells via the DLL4/Notch4/smad7 pathway.","date":"2019","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/31515487","citation_count":21,"is_preprint":false},{"pmid":"33145353","id":"PMC_33145353","title":"Amarogentin Inhibits Liver Cancer Cell Angiogenesis after Insufficient Radiofrequency Ablation via Affecting Stemness and the p53-Dependent VEGFA/Dll4/Notch1 Pathway.","date":"2020","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/33145353","citation_count":20,"is_preprint":false},{"pmid":"30691003","id":"PMC_30691003","title":"The PERK Branch of the Unfolded Protein Response Promotes DLL4 Expression by Activating an Alternative Translation Mechanism.","date":"2019","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/30691003","citation_count":20,"is_preprint":false},{"pmid":"34888208","id":"PMC_34888208","title":"DR-5 and DLL-4 mAb Functionalized SLNs of Gamma-Secretase Inhibitors- An Approach for TNBC Treatment.","date":"2020","source":"Advanced pharmaceutical bulletin","url":"https://pubmed.ncbi.nlm.nih.gov/34888208","citation_count":19,"is_preprint":false},{"pmid":"35602952","id":"PMC_35602952","title":"Engineered patterns of Notch ligands Jag1 and Dll4 elicit differential spatial control of endothelial sprouting.","date":"2022","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/35602952","citation_count":19,"is_preprint":false},{"pmid":"31034805","id":"PMC_31034805","title":"The bispecific antibody HB-32, blockade of both VEGF and DLL4 shows potent anti-angiogenic activity in vitro and anti-tumor activity in breast cancer xenograft models.","date":"2019","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/31034805","citation_count":19,"is_preprint":false},{"pmid":"22252294","id":"PMC_22252294","title":"Inhibition of Notch signaling by Dll4-Fc promotes reperfusion of acutely ischemic tissues.","date":"2012","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/22252294","citation_count":19,"is_preprint":false},{"pmid":"34858850","id":"PMC_34858850","title":"EMS1/DLL4-Notch Signaling Axis Augments Cell Cycle-Mediated Tumorigenesis and Progress in Human Adrenocortical Carcinoma.","date":"2021","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34858850","citation_count":18,"is_preprint":false},{"pmid":"35239998","id":"PMC_35239998","title":"Dysregulation of the miR-30c/DLL4 axis by circHIPK3 is essential for KSHV lytic replication.","date":"2022","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/35239998","citation_count":18,"is_preprint":false},{"pmid":"28946938","id":"PMC_28946938","title":"LDB2 regulates the expression of DLL4 through the formation of oligomeric complexes in endothelial cells.","date":"2018","source":"BMB reports","url":"https://pubmed.ncbi.nlm.nih.gov/28946938","citation_count":18,"is_preprint":false},{"pmid":"36050494","id":"PMC_36050494","title":"Affinity-matured DLL4 ligands as broad-spectrum modulators of Notch signaling.","date":"2022","source":"Nature chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/36050494","citation_count":17,"is_preprint":false},{"pmid":"33787845","id":"PMC_33787845","title":"Neuritin promotes angiogenesis through inhibition of DLL4/Notch signaling pathway.","date":"2021","source":"Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/33787845","citation_count":17,"is_preprint":false},{"pmid":"32899448","id":"PMC_32899448","title":"Endothelial Jagged1 Antagonizes Dll4/Notch Signaling in Decidual Angiogenesis during Early Mouse Pregnancy.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32899448","citation_count":17,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":51853,"output_tokens":11899,"usd":0.167022,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":23194,"output_tokens":4579,"usd":0.115223,"stage2_stop_reason":"end_turn"},"total_usd":0.282245,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"DLL4-Notch1 signaling between endothelial cells restricts tip-cell formation in response to VEGF, establishing the ratio between tip and stalk cells required for correct sprouting and branching. Genetic inactivation of one Dll4 allele or endothelial-specific Notch1 deletion increases tip cell numbers, while Notch activation reduces them, demonstrating lateral inhibition within angiogenic sprouts.\",\n      \"method\": \"Genetic mouse models (Dll4 haploinsufficiency, endothelial-specific Notch1 deletion), gamma-secretase inhibitors, soluble Jagged1 peptide treatment, retinal vascular imaging\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple genetic loss-of-function and pharmacological gain-of-function approaches in vivo, replicated across multiple labs\",\n      \"pmids\": [\"17259973\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Jagged1 antagonizes DLL4-Notch signaling in cells expressing Fringe family glycosyltransferases: upon Notch glycosylation by Fringe, DLL4-Notch signaling is enhanced while Jagged1 has weak signaling capacity and competes with DLL4, establishing that the equilibrium between two Notch ligands with opposing roles regulates angiogenesis.\",\n      \"method\": \"Genetic mouse models of Jagged1 and Dll4 manipulation, Fringe glycosyltransferase functional studies, in vivo retinal angiogenesis assays\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal genetic and biochemical approaches, published in high-impact journal, broadly replicated\",\n      \"pmids\": [\"19524514\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Neutralizing DLL4 with a selective antibody renders endothelial cells hyperproliferative and causes defective cell fate specification/differentiation both in vitro and in vivo, demonstrating that DLL4-mediated Notch signaling regulates endothelial cell proliferation and differentiation during active vascularization.\",\n      \"method\": \"DLL4-selective neutralizing antibody, in vitro endothelial cell assays, in vivo tumor xenograft models, histological analysis\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — antibody-based loss-of-function with multiple in vitro and in vivo readouts, replicated across tumor models\",\n      \"pmids\": [\"17183323\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Genetic experiments in postnatal mice reveal that the level of active Notch signaling is more important than direct DLL4-mediated cell-cell communication. Endothelial tip cells retain their function without Dll4 and are not replaced by adjacent Dll4-positive cells. Instead, Dll4-Notch signaling directs tip-derived endothelial cells into developing arteries, coupling sprouting angiogenesis and artery formation.\",\n      \"method\": \"Conditional genetic targeting of endothelial tip cells in vivo, mosaic Dll4 deletion, postnatal retinal angiogenesis analysis\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with cell-type-specific in vivo models and multiple phenotypic readouts\",\n      \"pmids\": [\"28714968\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Hypoxia via HIF-1alpha induces DLL4 expression and downstream Notch target genes Hey1 and Hey2. The activated Dll4-Notch-Hey2 signaling cascade leads to repression of COUP-TFII in endothelial progenitor cells, promoting arterial cell fate decision. Hey factors provide negative feedback by repressing HIF-1alpha-induced gene expression.\",\n      \"method\": \"Promoter analysis, HIF-1alpha induction in cell lines, endothelial progenitor cell culture, gene expression analysis\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — promoter analysis combined with cell-based functional assays, single lab\",\n      \"pmids\": [\"17045587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Beta-catenin (Wnt signaling) upregulates Dll4 transcription and strongly increases Notch signaling in endothelium, leading to functional and morphological vascular alterations including lack of vascular remodeling and loss of venous identity. Both in vivo and in vitro data establish a mechanistic link between Wnt and Notch signaling via DLL4.\",\n      \"method\": \"Endothelial-specific stabilization of beta-catenin in mice, in vitro endothelial cell assays, DLL4 promoter transcriptional analysis\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo and in vitro convergent evidence, single lab\",\n      \"pmids\": [\"20627076\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DLL4-mediated Notch signaling in tumor endothelium increases large vessel formation, reduces VEGFR2 expression in large blood vessels, decreases VEGFR3 overall, and decreases hypoxia-induced VEGF while increasing VEGFR1 in tumor stroma — multiple mechanisms by which DLL4-Notch confers resistance to bevacizumab (anti-VEGF) therapy.\",\n      \"method\": \"Retroviral DLL4 transduction of glioblastoma cells, tumor xenografts, bevacizumab treatment, gamma-secretase inhibitor treatment, molecular analysis of resistance mechanisms\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo tumor model with multiple molecular mechanistic readouts, single lab\",\n      \"pmids\": [\"21803743\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"DLL4 on dendritic cells is induced by pathogen-associated signals through TLR activation (but not by early inflammatory cytokines IL-1 and IL-18), and DLL4 signaling upregulates Rorc expression in T cells and directly targets Rorc and Il17 gene promoters to promote Th17 differentiation. DLL4 also inhibits Th2 cytokine production.\",\n      \"method\": \"In vitro T cell differentiation co-culture assays, siRNA knockdown, Notch signaling inhibition, gene promoter analysis\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — promoter analysis combined with functional differentiation assays, single lab\",\n      \"pmids\": [\"19494260\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DLL4-Notch1 signaling blockade disrupts postnatal lymphatic development by downregulating EphrinB2 expression (required for VEGFR3/VEGFC signaling), resulting in reduced lymphangiogenic sprouting, dilation of collecting lymphatic vessels with reduced mural cell coverage, and impaired wound healing/lymphangiogenesis.\",\n      \"method\": \"Function-blocking antibodies against Notch1 and Dll4 in mice, lymphatic development analysis, EphrinB2 expression analysis, wound healing model\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — antibody-based loss-of-function with multiple vascular phenotype readouts, single lab\",\n      \"pmids\": [\"21700774\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DLL4/Notch pathway regulates vessel regression by modulating vasoconstriction and blood flow: Dll4/Notch inhibition upregulates vasodilators (adrenomedullin) and suppresses vasoconstrictors (angiotensinogen), maintaining blood flow and preventing capillary regression. Angiotensin II induces rapid nonperfusion and regression of developing retinal capillaries.\",\n      \"method\": \"Genetic and pharmacologic Dll4/Notch inhibition in retinal oxygen-induced retinopathy model, Notch-regulated ankyrin repeat protein deletion, vasoactive molecule expression analysis\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and pharmacological approaches with mechanistic downstream pathway identification, single lab\",\n      \"pmids\": [\"21498671\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DLL4 expression in intestinal lacteals requires activation of VEGFR3 and VEGFR2, and genetic inactivation of Dll4 in lymphatic endothelial cells leads to lacteal regression and impaired dietary fat uptake, demonstrating a role for continuous DLL4 signaling in adult lymphatic vessel maintenance and function.\",\n      \"method\": \"Lymphatic endothelial cell-specific genetic inactivation of Dll4, VEGFR2/3 blocking experiments, dietary fat absorption assays, high-resolution intestinal stroma analysis\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — tissue-specific genetic deletion with functional dietary fat absorption readout and upstream receptor pathway identification\",\n      \"pmids\": [\"26529256\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Adhesion of endothelial cells to laminin-111 via alpha2beta1 and alpha6beta1 integrins triggers DLL4 expression, leading to subsequent Notch pathway activation. Foxc2 transcription is required but not sufficient for DLL4 induction. VEGF stimulates laminin gamma1 deposition, which leads to integrin signaling and DLL4 induction. Loss of integrins alpha2 or alpha6 mimics DLL4 silencing effects.\",\n      \"method\": \"siRNA knockdown of integrins, endothelial cell adhesion assays on laminin-111, 3D matrigel sprouting assay, VEGF stimulation experiments\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple siRNA knockdowns with functional sprouting assay readout, single lab\",\n      \"pmids\": [\"21474814\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Synaptojanin-2 binding protein (SYNJ2BP) physically interacts with the PDZ binding motif of DLL4 (and DLL1 but not Jagged-1), is preferentially expressed in stalk cells, enhances DLL4 protein stability, and promotes Notch signaling in endothelial cells. SYNJ2BP enables DLL4 interaction with Nectin-2 at adherens junctions.\",\n      \"method\": \"Co-immunoprecipitation, protein stability assays, siRNA knockdown, in vivo vascular density analysis in immunocompromised mice, in vitro endothelial cell assays\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding demonstrated by Co-IP with functional stability and signaling consequences, single lab\",\n      \"pmids\": [\"24025447\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"DLL4-containing exosomes can travel through 3D collagen matrix, transfer DLL4 protein to distant endothelial tip cells, activate Notch signaling in recipient cells, cause tip cell filopodia retraction, suppress sprout formation, increase endothelial cell motility, and suppress endothelial cell proliferation.\",\n      \"method\": \"3D microfluidic device, time-lapse confocal microscopy, exosome isolation and application, Notch signaling readouts\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct live imaging of exosome transfer with functional and signaling readouts, single lab\",\n      \"pmids\": [\"24504253\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"At limiting expression levels, DLL4 maintains the ability to inhibit B lineage choice and induce T lineage commitment at lower expression levels than DLL1. DLL4 expressed at physiological levels supports T lineage cells and is permissive for myeloid cells while still inhibiting B lymphopoiesis. These properties correlate with DLL4's more efficient induction of Notch target genes and inhibition of B/myeloid-specific transcription factors.\",\n      \"method\": \"OP9 stromal cell lines expressing incrementally discrete levels of Dll1 or Dll4, hematopoietic progenitor coculture, Notch target gene expression analysis, lineage output analysis\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — quantitative dose-response comparison between DLL1 and DLL4 with functional lineage commitment readout, single lab\",\n      \"pmids\": [\"20548034\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DLL4/Notch1 and BMP9/ALK1 signaling pathways are interdependent: canonical BMP9 signaling via ALK1-Smad1/5/9 is disrupted by inhibition of Notch signaling, and DLL4 activity is suppressed when the ALK1-Smad1/5/9 pathway is inhibited. BMP9/DLL4 synergistically induces complete endothelial quiescence requiring P27KIP1 and upregulation of thrombospondin-1.\",\n      \"method\": \"Human endothelial cell stimulation with BMP9 and DLL4, Notch and ALK1 inhibitors, Dll4+/- mouse vascular analysis, proteomics\",\n      \"journal\": \"Arteriosclerosis, thrombosis, and vascular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal pathway inhibition experiments with proteomics, in vitro and in vivo convergent data, single lab\",\n      \"pmids\": [\"26471266\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"DLL4 expression fluctuates in individual endothelial cells within sprouting vessels. High VEGF or DLL4 overexpression leads to Notch-dependent synchronization of DLL4 fluctuations within clusters, switching vessels from branching to expansion. Normal asynchronous Dll4 oscillations drive heterogeneity and branching, while synchronization drives vessel expansion.\",\n      \"method\": \"Live imaging in mouse retina in vivo, mouse embryonic stem cell-derived sprouting assays, DLL4 overexpression, Notch inhibition\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo and in vitro live imaging with genetic/pharmacological perturbations, single lab\",\n      \"pmids\": [\"27074663\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Leader cell identity during collective cell migration is dynamically regulated by DLL4 signaling through Notch1 and cellular stress. DLL4 is induced in leader cells after creation of a cell-free region, and leader cells are regulated via Notch1-DLL4 lateral inhibition. Mechanical stress inhibits DLL4 expression and leader cell formation.\",\n      \"method\": \"Single-cell gene expression analysis, computational modeling, time-lapse microscopy, Notch1-DLL4 inhibition\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — single-cell analysis combined with live imaging and perturbation experiments, single lab\",\n      \"pmids\": [\"25766473\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CCM1 silencing in endothelial cells causes decreased Notch3 activity in cocultured pericytes. Endothelial DLL4 stimulates Notch3 receptors on human brain pericytes; active Notch3 induces expression of PDGFRB2, N-Cadherin, HBEGF, TGFB1, NG2, and S1P genes, enhances pericyte adhesion to endothelial cells, limits pericyte migration/invasion, and enhances pericyte antiangiogenic function.\",\n      \"method\": \"Genetic manipulation of primary human endothelial cells and brain pericytes, Ccm1/Ccm2 endothelial-specific ablation in mouse models, Notch3 siRNA knockdown, pericyte-endothelial coculture\",\n      \"journal\": \"Stroke\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-type-specific genetic manipulation with functional coculture readouts and in vivo CCM mouse models, single lab\",\n      \"pmids\": [\"25791711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Multiple PDZ domain protein (MPDZ) physically interacts with the intracellular carboxyterminus of DLL4 (and DLL1) and enables their interaction with the adherens junction protein Nectin-2. MPDZ inactivation leads to impaired Notch signaling activity and increased blood vessel sprouting; tumor angiogenesis was enhanced upon endothelial-specific MPDZ inactivation.\",\n      \"method\": \"Co-immunoprecipitation, MPDZ gene inactivation in cellular models, embryonic mouse hindbrain vascular analysis, tumor angiogenesis models\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding demonstrated plus in vivo phenotype, single lab\",\n      \"pmids\": [\"29620522\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"LPA4/LPA6-mediated Gα12/Gα13-Rho-ROCK signaling activates YAP/TAZ in endothelial cells; YAP/TAZ knockdown increases β-catenin- and NICD-mediated endothelial DLL4 expression. LPA4/LPA6 or YAP/TAZ knockdown blocks EC sprouting, rescued by Notch inhibitor, demonstrating that LPA-YAP/TAZ signaling promotes angiogenesis by repressing DLL4.\",\n      \"method\": \"Endothelial-specific Lpa4;Lpa6 double knockout mice, siRNA knockdown of YAP/TAZ, fibrin gel sprouting assay, retinal angiogenesis analysis, Notch inhibition rescue experiments\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic double knockout with multiple in vitro and in vivo validation experiments, single lab\",\n      \"pmids\": [\"31335323\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Slug (SNAI2) transcription factor suppresses DLL4-Notch signaling in angiogenic endothelial cells, thereby promoting VEGFR2 expression. EC-specific Slug re-expression or reduced Notch signaling (gamma-secretase inhibition or Dll4 loss) rescues retinal angiogenesis in Slug knockout mice. Endothelial Slug is activated by SDF1alpha via CXCR4 and MAP kinase ERK5.\",\n      \"method\": \"Slug knockout mice, EC-specific re-expression, gamma-secretase inhibition, Dll4 loss-of-function, VEGF signaling inhibition, CXCR4/ERK5 pathway analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic rescue experiments with epistasis analysis and upstream pathway identification, single lab\",\n      \"pmids\": [\"33106502\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Indoxyl sulfate induces DLL4 protein expression in macrophages via inhibition of the ubiquitin-proteasome pathway through the deubiquitinating enzyme USP5, triggering Notch signaling. Macrophage uptake of indoxyl sulfate is mediated by OATP2B1; DLL4 antibody and OATP2B1 siRNA inhibit proinflammatory macrophage activation and atherosclerotic lesion development in mice.\",\n      \"method\": \"In vitro macrophage treatment with indoxyl sulfate, global proteomics, siRNA knockdown (macrophage-targeted lipid nanoparticles), 5/6 nephrectomy mouse model, Dll4 antibody treatment, Ldlr-/- atherosclerosis model\",\n      \"journal\": \"Circulation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — proteomics-guided mechanistic validation with in vivo macrophage-targeted siRNA and antibody approaches, single lab\",\n      \"pmids\": [\"30586693\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In the context of diabetic wounds, high glucose levels activate a positive Dll4-Notch1 feedback loop, and Notch1 inactivation specifically in keratinocytes cancels the repressive effects of this loop on wound healing in diabetes, demonstrating that the Dll4-Notch1 loop in keratinocytes impairs diabetic wound healing.\",\n      \"method\": \"Genetic loss-of-function in diabetic mouse models, keratinocyte-specific Notch1 inactivation, local Notch signaling inhibition, wound healing analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-type-specific genetic loss-of-function with functional wound healing readout, single lab\",\n      \"pmids\": [\"30886104\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Both antibody-based and genetic inactivation of Dll4 or Notch1 induces hyperpermeability in arterial endothelial cells by increasing transcytosis without junctional destabilization. Endothelial Sox17 deletion represses Dll4 in retinal arteries, phenocopying Dll4 blockade. Dll4 blocking activates SREBP1-mediated lipogenic transcription and caveolae formation. Inhibition of SREBP1 or VEGF-VEGFR2 attenuates Dll4 blockade-driven retinal leakage, establishing a Sox17-Dll4-SREBP1 axis controlling transcytosis independently of tight junctions.\",\n      \"method\": \"Antibody-based and genetic Dll4/Notch1 inactivation, Sox17 endothelial deletion, transcriptomic profiling, SREBP1 inhibition, VEGFR2 inhibition, hypertension-induced retinal edema model\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal genetic and pharmacological approaches identifying a novel transcytosis mechanism with pathway validation, single rigorous study\",\n      \"pmids\": [\"32078435\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In atrophic conditions, microvascular endothelium upregulates and releases the Notch ligand DLL4, which activates muscular Notch2 without direct cell-cell contact. Inhibition of the Dll4-Notch2 axis substantially prevents disuse- or diabetes-induced muscle atrophy and promotes overloading-induced muscle hypertrophy in mice, establishing an endothelial-to-muscle signaling axis controlling skeletal muscle mass.\",\n      \"method\": \"Myofiber Notch2 conditional knockout, Dll4 inhibition, disuse and diabetic atrophy mouse models, mechanical overload model, DLL4/Notch2 axis analysis\",\n      \"journal\": \"Nature metabolism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — tissue-specific genetic loss-of-function with multiple in vivo functional readouts identifying a novel inter-cellular signaling mechanism\",\n      \"pmids\": [\"35228746\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DLL4 is expressed in a sub-population of bipotent hematoendothelial progenitors (HEPs) in hESCs and segregates their hematopoietic versus endothelial potential. DLL4-high HEPs are enriched for endothelial potential, while DLL4-low/-negative HEPs are committed to hematopoietic lineage. DLL4 stimulation enhances hematopoietic differentiation of HEPs and increases clonogenic hematopoietic progenitors.\",\n      \"method\": \"hESC differentiation, clonal analysis, transcriptome analysis, confocal microscopy of embryoid bodies, DLL4 stimulation of HEPs\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clonal and transcriptome analysis with functional stimulation experiments, single lab\",\n      \"pmids\": [\"25778099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Dll4 acts as a negative regulator of intra-aortic hematopoietic cluster (IAHC) recruitment: blocking Dll4 promotes entrance of new hemogenic Gfi1+ cells into IAHC and increases the number of cells acquiring HSC activity. IACHs form through a two-step process where Dll4 inhibits the recruitment phase.\",\n      \"method\": \"Live imaging of organotypic slice cultures, clonal analysis, mathematical modeling, Dll4 blocking experiments in mouse embryos\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live imaging combined with clonal analysis and mathematical modeling, single lab\",\n      \"pmids\": [\"32149421\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"DLL4 preferentially activates NOTCH1 over NOTCH2, whereas DLL1 is equally effective in activating NOTCH1 and NOTCH2. The discriminating potential lies in the region between the N-terminus and EGF repeat three of the ligand ectodomain. The ectodomains dictate selective ligand function in vivo during somitogenesis.\",\n      \"method\": \"Cellular co-culture signaling assays, biochemical binding studies, chimeric ligand knock-in mouse models, somitogenesis and myogenesis analysis\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — biochemical binding studies, chimeric ligand functional assays, and in vivo genetic validation in multiple contexts\",\n      \"pmids\": [\"30289388\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DLL4, but not DLL1, is an efficient cis-inhibitor of Notch signaling, causing reduced net activation of Notch in cells co-expressing ligand and receptor. This differential cis-inhibitory property contributes to context-dependent functional divergence between DLL1 and DLL4.\",\n      \"method\": \"Conditional overexpression from same genomic locus (Hprt), Dll1Dll4 knock-in mice, in vitro Notch signaling assays for cis-inhibition and trans-activation\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro signaling assays combined with in vivo genetic models, single lab\",\n      \"pmids\": [\"26114479\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"LPS/TLR4 signaling induces ERK phosphorylation, which causes FOXC2-ERK protein ligation and ERK-dependent FOXC2 serine/threonine phosphorylation, subsequently activating DLL4 gene expression. FOXC2 binds to the DLL4 promoter in vivo. ERK inhibition or FOXC2 siRNA attenuates LPS-induced DLL4 expression and angiogenic sprouting.\",\n      \"method\": \"TLR4/LPS stimulation in human lung endothelial cells, ERK inhibition, ERK-2 dominant negative transfection, FOXC2-siRNA, in vivo mouse lung analysis, FOXC2+/- mice\",\n      \"journal\": \"The Journal of physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — promoter binding combined with multiple loss-of-function approaches in vitro and in vivo, single lab\",\n      \"pmids\": [\"29380370\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The DLL4 5'-UTR harbors an Internal Ribosomal Entry Site (IRES) that is efficiently utilized during hypoxia and ER stress (cap-independent translation). PERK kinase (activated by ER stress) drives DLL4 IRES-mediated translation, and hnRNP-A1 acts as an IRES-Trans-Acting Factor (ITAF) participating in IRES-dependent DLL4 translation during ER stress.\",\n      \"method\": \"IRES reporter assays, PERK inhibition/activation, hnRNP-A1 characterization, hypoxia and ER stress treatment of cells\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reporter assays for IRES activity combined with identification of specific trans-acting factors, single lab\",\n      \"pmids\": [\"30691003\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"KSHV vGPCR upregulates DLL4 through an ERK-dependent mechanism in lymphatic endothelial cells, while vFLIP induces JAG1 through NFkappaB-dependent signaling. Both ligands signal through NOTCH4 and suppress cell cycle genes in adjacent lymphatic endothelial cells to induce quiescence.\",\n      \"method\": \"KSHV gene expression in lymphatic endothelial cells, ERK and NFkappaB pathway inhibition, gene expression profiling, functional Notch signaling assays\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — specific viral gene attribution with pathway inhibitor validation and gene expression profiling, single lab\",\n      \"pmids\": [\"19816565\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TMZ treatment promotes nuclear translocation of MMP14 followed by extracellular release of DLL4. Released DLL4 stimulates cleavage of Notch3, its nuclear translocation, and induction of sphering capacity and stemness in glioblastoma cells.\",\n      \"method\": \"Multiple PDX GBM models and glioma cell lines, MMP14 expression/localization analysis, Kiloplex ELISA-based protein array, DLL4/Notch3 functional studies\",\n      \"journal\": \"International journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic identification of MMP14-DLL4-Notch3 axis with multiple models, single lab\",\n      \"pmids\": [\"31443114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Soluble DLL4 activates Notch signaling in endothelial cells, increases VE-cadherin expression at intercellular junctions (but not ZO-1), and decreases vascular permeability. This permeability reduction acts through a cAMP/PKA pathway: PKA inhibition reverses the DLL4-mediated permeability reduction and reduces Hey1 expression. PKA knockdown reduces VE-cadherin junctional expression.\",\n      \"method\": \"Recombinant soluble DLL4 treatment of EC monolayers, gamma-secretase inhibitor, PKA inhibitors, hydraulic conductivity in rat mesenteric microvessels in vivo, FITC-albumin permeability assays\",\n      \"journal\": \"American journal of physiology. Heart and circulatory physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro and in vivo vascular permeability assays with pharmacological pathway dissection, single lab\",\n      \"pmids\": [\"30681366\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In zebrafish valvulogenesis, blood flow activates Notch signaling, which drives lateral inhibition between endocardial cells mediated by DLL4. DLL4-positive endocardial cells ingress into the cardiac jelly to form an abluminal cell population in response to Wnt9a (produced via Erk5-Klf2-Wnt9a cascade). Mechanical stimulation activates parallel mechanosensitive signaling pathways (Notch and Klf2) that intersect to drive valve formation.\",\n      \"method\": \"Zebrafish genetic models, live imaging, DLL4-positive cell lineage tracking, Notch and Wnt9a pathway perturbation experiments\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis in zebrafish with live imaging validation, single lab\",\n      \"pmids\": [\"34610316\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Pre-existing embryonic coronary plexus expresses DLL4, and DLL4-NOTCH1 signaling mediates angiogenic expansion of this plexus to vascularize the expanding myocardium in neonates, and also revascularizes the regenerating neonatal heart. Ventricular endocardial cells do not contribute to new coronary vessels.\",\n      \"method\": \"Lineage-tracing, gain- and loss-of-function genetic experiments in mice, neonatal heart regeneration model\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — convergent lineage tracing and genetic gain/loss-of-function establishing DLL4-NOTCH1 as the mechanistic driver, single rigorous study\",\n      \"pmids\": [\"34497373\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Coronary arterial development requires a DLL4-Jag1-EphrinB2 signaling cascade: endocardial Jag1 removal blocks sinus venosus capillary sprouting, while Dll4 inactivation stimulates excessive capillary growth. Forced Dll4 expression or Mfng (glycosyltransferase) blocks coronary plexus remodeling and arterial differentiation. EphrinB2 is a critical effector of antagonistic Dll4 and Jag1 functions in arterial morphogenesis.\",\n      \"method\": \"Endocardial Jag1 conditional deletion, Dll4 inactivation, forced Dll4/Mfng expression, Efnb2 endocardial deletion, angiogenic rescue experiments in ventricular explants and human endothelial cells\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple genetic loss- and gain-of-function with mechanistic rescue experiments in vivo and in vitro, single rigorous study\",\n      \"pmids\": [\"31789590\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"LDB2 (LIM-domain binding protein 2) regulates basal and VEGF-induced DLL4 expression in endothelial cells by binding to the DLL4 promoter region through oligomeric complexes with LMO2/TAL1/GATA2. LDB2 overexpression increases DLL4 expression; LDB2 knockdown decreases DLL4 expression and enhances endothelial sprouting.\",\n      \"method\": \"siRNA knockdown and overexpression of LDB2, DLL4 promoter binding assays, zebrafish ldb2-morpholino, in vitro sprouting/tubular network assays\",\n      \"journal\": \"BMB reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — promoter binding assays combined with loss- and gain-of-function and in vivo zebrafish model, single lab\",\n      \"pmids\": [\"28946938\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Affinity-matured DLL4 variant (DeltaMAX) binds human and murine Notch receptors with 500- to 1000-fold increased affinity compared to wild-type human DLL4. DeltaMAX potently activates Notch in plate-bound, bead-bound, and cellular formats. As a soluble decoy, DeltaMAX inhibits Notch in reporter and neuronal differentiation assays, demonstrating dual agonist/antagonist utility.\",\n      \"method\": \"Protein engineering/affinity maturation, in vitro Notch activation assays (plate-bound, bead-bound, cellular), reporter assays, neuronal differentiation assays, T cell stimulation assays\",\n      \"journal\": \"Nature chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with quantitative binding measurements and multiple functional validation assays\",\n      \"pmids\": [\"36050494\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Epsin1 modulates the sorting of DLL4 into exosomes from tubular epithelial cells under high glucose conditions. Exosomes enriched with DLL4 are captured by macrophages and promote M1 macrophage activation via Notch1 (N1ICD) activation. Epsin1 knockdown reduces DLL4 in TEC-exosomes and inhibits macrophage N1ICD activation and iNOS expression.\",\n      \"method\": \"Mass spectrometry of urine exosomes, Epsin1 siRNA knockdown, in vitro THP-1 macrophage treatment with exosomes, in vivo C57BL/6 mouse model, western blot, db/db diabetic mice\",\n      \"journal\": \"Molecular therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic identification of Epsin1-mediated DLL4 exosomal sorting with in vitro and in vivo validation, single lab\",\n      \"pmids\": [\"37016580\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"MEDI0639 (anti-DLL4 antibody) inhibits the binding of Notch1 to DLL4 via a novel epitope not previously described, reversing Notch1-mediated suppression of HUVEC growth in vitro. MEDI0639 promotes tubule formation in 3D endothelial outgrowth assay (disrupting Dll4-Notch axis), but inhibits tubule formation in 2D endothelial-fibroblast coculture. In vivo, MEDI0639 promotes human vessel formation and reduces mural cell coverage.\",\n      \"method\": \"Notch1-DLL4 binding inhibition assays, HUVEC growth assays, 3D and 2D angiogenesis assays, in vivo human endothelial cell angiogenesis assay in mice\",\n      \"journal\": \"Molecular cancer therapeutics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding inhibition combined with multiple in vitro and in vivo functional assays, single lab\",\n      \"pmids\": [\"22679110\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Palmitic acid induces macrophage DLL4 signaling, which in turn triggers senescence in vascular smooth muscle cells, reducing collagen synthesis and deposition, thus promoting atherosclerotic plaque instability. Macrophage-specific DLL4 knockout in atherosclerotic mice leads to reduced plaque burden and improved stability.\",\n      \"method\": \"Human cohort studies, macrophage-specific DLL4 knockout in atherosclerotic mouse models, palmitic acid treatment of macrophages, vascular smooth muscle cell senescence assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — macrophage-specific genetic knockout with mechanistic cellular downstream identification, single lab\",\n      \"pmids\": [\"38346959\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Dll4-Notch signaling in DLL4-expressing cancer cells (SCLC) plays a critical role in liver metastasis by regulating NF-κB signaling. Dll4-Fc (soluble DLL4) acts as a blocker, and Dll4-Fc-expressing cancer cells show downregulated NF-κB activities (both classical and alternative pathways) by reducing Notch1 signaling, resulting in reduced liver metastasis.\",\n      \"method\": \"Soluble Dll4-Fc generation, SCLC cell line transduction, mouse liver metastasis model, PCR array analysis, electrophoretic mobility shift assay for NF-κB activity, Notch1 signaling analysis\",\n      \"journal\": \"Molecular cancer therapeutics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional metastasis model combined with NF-κB pathway analysis, single lab\",\n      \"pmids\": [\"22989420\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Dll4 blockade (pharmacological or genetic) induces accumulation of thymic dendritic cells and CD4+CD25+FoxP3+ regulatory T cells in the thymic cortex. Dll4 blockade converts DN1 T cell progenitors to immature DCs that induce Treg differentiation through a Flt3-independent, DC-dependent mechanism requiring MHC II expression. This mechanism depends on transcriptional upregulation of PU.1, Irf-4, Irf-8, and CSF-1.\",\n      \"method\": \"Pharmacological Dll4 blockade, genetic inactivation models, thymectomy experiments, DC-T cell coculture, anti-Dll4 antibody treatment in type 1 diabetes model\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and pharmacological loss-of-function with epistasis analysis, single lab\",\n      \"pmids\": [\"22547652\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"VEGF165 inhibits pro-fibrotic differentiation of endometrial stromal cells via the DLL4/Notch4/Smad7 pathway; inhibiting Smad7 or Notch4 blocks the anti-fibrotic effect of VEGF165, demonstrating that DLL4 and Notch4 are essential downstream molecules for VEGF165's anti-fibrotic function.\",\n      \"method\": \"VEGF165 treatment of human primary endometrial stromal cells, conditional VEGF reduction in mice, Smad7 and Notch4 inhibition, TGFbeta1-induced fibrosis model\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple inhibitor experiments establishing epistatic relationship, single lab\",\n      \"pmids\": [\"31515487\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DLL4 is a transmembrane Notch ligand expressed predominantly in arterial and tip-cell endothelium that signals in trans through Notch1 (and Notch2/3/4 in specific contexts) to laterally inhibit tip-cell formation and restrict angiogenic sprouting in response to VEGF; its expression is regulated by VEGF, HIF-1alpha, Wnt/beta-catenin, integrin-laminin, Fringe-mediated glycosylation (which modulates its competitive balance with Jagged1), and transcriptional regulators including FOXC2, LDB2, and Slug, while its protein levels are stabilized by SYNJ2BP and MPDZ at adherens junctions; downstream, DLL4-Notch signaling controls arterio-venous specification, lymphatic maintenance, transcytosis-dependent vascular barrier function (via Sox17-SREBP1), endothelial quiescence (in cooperation with BMP9/ALK1 and via P27KIP1), and skeletal muscle mass (through an endothelial-to-myofiber Dll4-Notch2 axis), as well as T-cell lineage commitment in the thymus and hematopoietic progenitor fate decisions.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"DLL4 is a transmembrane Notch ligand that operates as the central rheostat of sprouting angiogenesis, signaling in trans through Notch1 to enforce lateral inhibition that fixes the ratio of tip to stalk cells during VEGF-driven vascular outgrowth [#0, #2]. Within this circuit DLL4 expression fluctuates in individual endothelial cells, and the synchronization versus asynchrony of these oscillations toggles vessels between expansion and branching [#16]; the integrated level of Notch activity, more than direct cell-cell contact, determines outcome and channels tip-derived cells into arteries [#3]. DLL4 selectively activates NOTCH1 over NOTCH2 through its N-terminal ectodomain and, unlike DLL1, acts as an efficient cis-inhibitor, properties that account for its distinct functional output [#28, #29]. Its signaling strength is set by a competitive balance with Jagged1 that is tuned by Fringe glycosyltransferases [#1, #37]. DLL4 transcription is driven by a convergent set of inputs—VEGF/HIF-1alpha-arterial programs [#4], Wnt/beta-catenin [#5], integrin-laminin adhesion acting through FOXC2 [#11, #30], and LDB2/LMO2/TAL1/GATA2 complexes [#38]—and is repressed by Slug and by YAP/TAZ [#21, #20]; protein levels are further controlled post-transcriptionally by IRES-mediated translation under hypoxia/ER stress [#31], by deubiquitination via USP5 [#22], and by junctional stabilization through the PDZ proteins SYNJ2BP and MPDZ that link DLL4 to Nectin-2 at adherens junctions [#12, #19]. Through these mechanisms DLL4-Notch controls arterio-venous and coronary vascular morphogenesis [#36, #37], lymphatic sprouting and lacteal maintenance via EphrinB2 and VEGFR3 [#8, #10], endothelial quiescence in cooperation with BMP9/ALK1 and P27KIP1 [#15], and vascular barrier function through a Sox17-Dll4-SREBP1 axis that limits transcytosis [#24]. Beyond the vasculature DLL4 governs T-cell lineage commitment and thymic Treg generation [#14, #44], hematoendothelial fate decisions [#26, #27], and an endothelial-to-myofiber Dll4-Notch2 axis controlling skeletal muscle mass [#25]; it is also exploited in tumor angiogenesis and metastasis and in macrophage-driven atherosclerosis [#2, #6, #42].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Establishing whether DLL4 is required for endothelial differentiation versus proliferation defined its core role as a brake on uncontrolled vascular growth.\",\n      \"evidence\": \"DLL4-selective neutralizing antibody with in vitro endothelial assays and tumor xenografts\",\n      \"pmids\": [\"17183323\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve which Notch receptor mediates the effect\", \"Mechanism of fate specification downstream of Notch not defined\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Resolving how endothelial sprouts select tip versus stalk identity showed DLL4-Notch1 lateral inhibition sets the tip:stalk ratio in response to VEGF.\",\n      \"evidence\": \"Dll4 haploinsufficiency, endothelial Notch1 deletion, gamma-secretase inhibitors and retinal imaging in mice\",\n      \"pmids\": [\"17259973\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not explain how DLL4 levels are dynamically encoded across the sprout\", \"Transcriptional control of DLL4 by VEGF left unaddressed\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defining why two Notch ligands have opposing angiogenic roles revealed that Fringe glycosylation tilts a DLL4-versus-Jagged1 competitive equilibrium.\",\n      \"evidence\": \"Genetic Jagged1/Dll4 manipulation and Fringe glycosyltransferase studies in mouse retinal angiogenesis\",\n      \"pmids\": [\"19524514\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quantitative thresholds of ligand balance not defined\", \"Structural basis of Fringe-dependent discrimination unresolved\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Identifying upstream transcriptional drivers connected angiogenic signaling and Wnt to DLL4, and quantified DLL4's distinct potency in lineage commitment.\",\n      \"evidence\": \"Endothelial beta-catenin stabilization in mice plus DLL4 promoter analysis; OP9 dose-titration cocultures comparing DLL1 and DLL4\",\n      \"pmids\": [\"20627076\", \"20548034\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct beta-catenin promoter occupancy not fully resolved\", \"Molecular basis of DLL4's greater potency versus DLL1 not yet mapped\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Mapping additional inducers and downstream effectors extended DLL4 control to integrin-laminin adhesion, lymphatic development, and vessel regression.\",\n      \"evidence\": \"Integrin siRNA with laminin adhesion and 3D sprouting assays; anti-Dll4/Notch1 antibodies in lymphatic and oxygen-induced retinopathy models\",\n      \"pmids\": [\"21474814\", \"21700774\", \"21498671\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"FOXC2 shown necessary but not sufficient for DLL4 induction\", \"Link between EphrinB2 regulation and VEGFR3 signaling mechanistically incomplete\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Discovering DLL4 PDZ-motif binding partners explained how DLL4 protein is stabilized and positioned at adherens junctions to sustain Notch output.\",\n      \"evidence\": \"Co-IP, protein stability assays and siRNA in endothelial cells (SYNJ2BP); later MPDZ Co-IP plus in vivo hindbrain and tumor angiogenesis\",\n      \"pmids\": [\"24025447\", \"29620522\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab Co-IP without independent reciprocal validation for each partner\", \"How junctional localization tunes trans-signaling strength not quantified\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"A series of studies expanded DLL4 into adult homeostasis—lacteal maintenance, endothelial quiescence, pericyte regulation, and hematoendothelial fate.\",\n      \"evidence\": \"Lymphatic-specific Dll4 deletion with fat-absorption assays; BMP9/ALK1 reciprocal inhibition with proteomics; Notch3 pericyte cocultures; hESC clonal differentiation\",\n      \"pmids\": [\"26529256\", \"26471266\", \"25791711\", \"25778099\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interdependence of DLL4 and BMP9/ALK1 mechanistically partial\", \"Context-specificity of which Notch receptor DLL4 engages not uniformly resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Live imaging of DLL4 dynamics reframed angiogenic patterning as an oscillatory system in which synchronization switches branching to expansion.\",\n      \"evidence\": \"In vivo retinal live imaging and ES-cell sprouting assays with DLL4 overexpression and Notch inhibition\",\n      \"pmids\": [\"27074663\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular clock generating DLL4 oscillations not identified\", \"Single-lab observation\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Testing whether direct DLL4 contact or net Notch level dominates showed integrated Notch activity directs tip-derived cells into arteries.\",\n      \"evidence\": \"Mosaic Dll4 deletion and tip-cell genetic targeting in postnatal mouse retina\",\n      \"pmids\": [\"28714968\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How tip cells retain function without Dll4 not fully explained\", \"Compensating ligand sources not defined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Biochemical and genetic dissection of the ectodomain established the molecular basis of DLL4 receptor selectivity and additional transcriptional regulators.\",\n      \"evidence\": \"Chimeric ligand knock-in mice and binding studies (NOTCH1 vs NOTCH2 selectivity); LDB2 promoter complexes; LPS-FOXC2-ERK promoter axis\",\n      \"pmids\": [\"30289388\", \"28946938\", \"29380370\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural detail of the discriminating N-terminal region not solved\", \"Combinatorial logic among transcriptional inputs unresolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identifying the Sox17-Dll4-SREBP1 axis revealed a transcytosis-based, junction-independent mechanism by which DLL4 maintains the vascular barrier.\",\n      \"evidence\": \"Antibody and genetic Dll4/Notch1 inactivation, Sox17 deletion, transcriptomics and SREBP1 inhibition in retinal models\",\n      \"pmids\": [\"32078435\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct molecular link from NICD to SREBP1 activation not fully traced\", \"Caveolae-formation step mechanistically incomplete\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrating an endothelial-to-myofiber Dll4-Notch2 axis and engineering high-affinity DLL4 variants extended DLL4 biology beyond the vasculature and into reagent design.\",\n      \"evidence\": \"Myofiber Notch2 conditional knockout with atrophy/overload models; affinity-matured DeltaMAX variant with quantitative binding and functional assays\",\n      \"pmids\": [\"35228746\", \"36050494\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How released DLL4 acts without cell-cell contact mechanistically open\", \"In vivo therapeutic translation of engineered variants not established\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Disease-context studies positioned DLL4 in macrophage-driven atherosclerosis and exosome-mediated intercellular Notch signaling.\",\n      \"evidence\": \"Macrophage-specific DLL4 knockout in atherosclerotic mice; Epsin1-dependent DLL4 exosome sorting with macrophage activation assays\",\n      \"pmids\": [\"38346959\", \"37016580\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which DLL4 triggers VSMC senescence partial\", \"Single-lab disease-context findings\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis and quantitative thresholds that convert DLL4 oscillation, cis-inhibition, and ligand competition into specific cell-fate outputs across tissues remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model linking DLL4 expression dynamics to receptor selection in vivo\", \"Mechanism of contact-independent (soluble/exosomal) DLL4 signaling incompletely defined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [0, 2, 28, 39]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 1, 28, 29]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [1, 29]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 28]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [13, 40]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [33, 34, 43]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 28]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [3, 36, 37]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [7, 14, 44]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"NOTCH1\", \"NOTCH2\", \"NOTCH3\", \"NOTCH4\", \"SYNJ2BP\", \"MPDZ\", \"JAG1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}