{"gene":"DLL4","run_date":"2026-04-28T17:46:02","timeline":{"discoveries":[{"year":2009,"finding":"Jagged1 antagonizes DLL4-Notch signaling in endothelial cells expressing Fringe family glycosyltransferases; glycosylation of Notch enhances DLL4-Notch signaling whereas Jagged1 has weak signaling capacity and competes with DLL4, establishing opposing roles for the two ligands in angiogenesis.","method":"Genetic mouse models, in vivo angiogenesis assays, functional studies with Fringe glycosyltransferases","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal genetic and biochemical approaches, highly cited foundational paper","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, demonstrating that DLL4-mediated Notch signaling regulates endothelial cell proliferation and differentiation and is crucial during active vascularization.","method":"Anti-DLL4 neutralizing antibody treatment, in vitro and in vivo endothelial cell assays, tumor models","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 — multiple in vitro and in vivo models, highly cited foundational paper","pmids":["17183323"],"is_preprint":false},{"year":2007,"finding":"DLL4 expression is dynamically induced by VEGF in the retinal vasculature and acts as a negative feedback regulator to prevent overexuberant angiogenic sprouting; pharmacological inhibition of DLL4/Notch signaling (via soluble DLL4-Fc or blocking antibody) produces enhanced sprouting and increased endothelial proliferation.","method":"Intraocular administration of soluble DLL4-Fc and blocking antibody; Dll4 haploinsufficiency mouse model; postnatal retinal vascular analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — genetic and pharmacological loss-of-function with defined vascular phenotypes, replicated by multiple approaches","pmids":["17296940"],"is_preprint":false},{"year":2003,"finding":"VEGF, but not bFGF, induces DLL4 and Notch1 gene expression in human arterial endothelial cells via VEGFR1 and VEGFR2 through a PI3K/Akt pathway, independently of MAPK and Src; constitutive Notch activation stabilizes endothelial network formation.","method":"Gene expression analysis, pharmacological inhibitors of PI3K/Akt, MAPK, and Src; 3D angiogenesis model; Matrigel network formation assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — multiple signaling pathway inhibitors used, orthogonal functional assays, highly cited","pmids":["12482957"],"is_preprint":false},{"year":2011,"finding":"Simultaneous genetic inactivation of Dll1 and Dll4 in mouse intestinal epithelium causes complete conversion of proliferating progenitors into postmitotic goblet cells with loss of intestinal stem cells, establishing that DLL1 and DLL4 together are the physiological Notch ligands required for intestinal stem cell maintenance.","method":"Inducible gut-specific gene targeting (Vil-Cre-ERT2) in mice; single and double conditional knockouts; lineage analysis with Notch1 reporter","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 2 — clean genetic epistasis with specific stem cell phenotype, multiple mouse models","pmids":["21238454"],"is_preprint":false},{"year":2017,"finding":"Genetic experiments in postnatal mice reveal that the level of active Notch signaling (not direct DLL4-mediated cell-cell communication per se) is the key determinant of vessel growth; Notch activation directs tip-derived endothelial cells into developing arteries, coupling sprouting angiogenesis with artery formation. Endothelial VEGF-A and CXCR4 expression are key processes controlling Notch-dependent vessel growth.","method":"Endothelial-specific genetic targeting of Dll4 in tip cells in postnatal mice; conditional knockout models; retinal vascular analysis","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 — rigorous in vivo genetic epistasis, multiple conditional mouse models","pmids":["28714968"],"is_preprint":false},{"year":2010,"finding":"Endothelial-specific stabilization of Wnt/β-catenin signaling upregulates Dll4 transcription and strongly increases Notch signaling in the endothelium, linking Wnt and Notch signaling pathways in vascular development and arteriovenous specification.","method":"Endothelial-specific β-catenin gain-of-function mouse models; in vitro β-catenin activation; chromatin/transcriptional analysis of Dll4 promoter","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 — both in vivo and in vitro evidence with genetic models; mechanistic link demonstrated","pmids":["20627076"],"is_preprint":false},{"year":2008,"finding":"Foxc1 and Foxc2 transcription factors directly activate the Dll4 promoter and the Notch target Hey2 promoter via Foxc binding elements; Foxc2 physically interacts with a Notch transcriptional activation complex (Su(H)/NICD) to induce Hey2 promoter activity; VEGF-activated PI3K and ERK pathways modulate Foxc transcriptional activity in Dll4 and Hey2 induction.","method":"Promoter reporter assays, co-immunoprecipitation, siRNA knockdown, VEGF stimulation with PI3K/ERK inhibitors in endothelial cells","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 1-2 — direct protein-protein interaction demonstrated by Co-IP; promoter functional validation by mutagenesis/binding","pmids":["18545664"],"is_preprint":false},{"year":2013,"finding":"Arterial Dll4 expression is regulated combinatorially by Notch signaling (via RBPJ/NICD direct binding) and SoxF transcription factors (Sox7, Sox18) through two arterial-specific enhancers; combinatorial loss of both SoxF and RBPJ binding ablates all Dll4 enhancer activity and results in loss of arterial markers and dorsal aorta.","method":"Arterial-specific enhancer characterization in mouse and zebrafish; transgenic reporter assays; combined Sox7/Sox18/Rbpj knockdown; endogenous dll4 expression analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 — enhancer characterization with mutagenesis, genetic epistasis in two model organisms","pmids":["23818617"],"is_preprint":false},{"year":2006,"finding":"Hypoxia induces DLL4 expression through HIF-1α, which leads to activation of Notch target genes Hey1 and Hey2; in endothelial progenitor cells, hypoxia-mediated upregulation of DLL4 and Hey2 represses COUP-TFII (a venous identity regulator), promoting arterial cell fate; Hey factors create a negative feedback on HIF-1α-induced gene expression.","method":"Promoter analysis, HIF-1α overexpression/knockdown, endothelial progenitor cell culture under hypoxia, reporter assays","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 — promoter analysis and functional cell-based assays, single lab","pmids":["17045587"],"is_preprint":false},{"year":2012,"finding":"The microRNA-30 family directly targets the DLL4 3'UTR to repress DLL4 expression; miR-30b overexpression in endothelial cells increases vessel number/length in sprouting assays; microinjection of miR-30 mimics in zebrafish suppresses dll4 and causes excessive intersegmental vessel sprouting; target protector against the miR-30 site in dll4 3'UTR upregulates dll4.","method":"miRNA target site mutagenesis/target protector, overexpression in endothelial cells, zebrafish microinjection, sprouting angiogenesis assay","journal":"Blood","confidence":"High","confidence_rationale":"Tier 1-2 — target protector validation in vivo and in vitro; multiple orthogonal methods","pmids":["23086751"],"is_preprint":false},{"year":2015,"finding":"DLL4 expression in intestinal lacteals requires VEGFR3 and VEGFR2 activation; genetic inactivation of Dll4 specifically in lymphatic endothelial cells leads to lacteal regression and impaired dietary fat uptake, establishing DLL4 as a regulator of adult lymphatic vessel maintenance and intestinal fat absorption.","method":"Lymphatic endothelial cell-specific Dll4 conditional knockout mice; VEGFR inhibition; dietary fat uptake assays; histological analysis","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — tissue-specific conditional knockout with defined functional phenotype","pmids":["26529256"],"is_preprint":false},{"year":2015,"finding":"DLL4 is expressed on bone marrow osteocalcin-positive (Ocn+) mesenchymal cells; selective depletion of DLL4 from these cells recapitulates thymopoietic abnormality (reduced thymus-seeding progenitors and T cell generation), establishing that bone marrow DLL4 drives thymus-seeding progenitor generation.","method":"In vivo deletion of DLL4 from Ocn+ cells; conditional cell-specific knockouts; progenitor frequency analysis; thymic function assays","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 — cell-specific in vivo deletion with defined cellular phenotype","pmids":["25918341"],"is_preprint":false},{"year":2011,"finding":"Notch1-Dll4 signaling regulates postnatal lymphatic development; antibody blockade of Notch1 and Dll4 results in defective lymphatic sprouting associated with downregulation of EphrinB2 (which mediates VEGFR3/VEGFC signaling), dilation of collecting lymphatics with reduced mural cell coverage, and impaired wound-associated lymphangiogenesis.","method":"Function-blocking antibodies against Notch1 and Dll4 in mice; EphrinB2 expression analysis; wound healing lymphangiogenesis model","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 — pharmacological and pathway-level mechanistic analysis in vivo","pmids":["21700774"],"is_preprint":false},{"year":2009,"finding":"DLL4 expression in endothelial cells of the tumor microenvironment activates Notch3 signaling in co-cultured T-ALL tumor cells, promoting their escape from dormancy; neutralization of DLL4 greatly reduces EC-mediated Notch3 activation in T-ALL cells and blocks tumorigenesis.","method":"EC-tumor cell co-culture, angiogenic factor stimulation, DLL4 neutralization, Notch3 silencing by RNAi, in vivo tumorigenicity assays","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — co-culture and RNAi mechanistic evidence, single lab","pmids":["19208840"],"is_preprint":false},{"year":2011,"finding":"Adhesion of endothelial cells to laminin-111 via α2β1 and α6β1 integrins triggers DLL4 expression and subsequent Notch pathway activation; VEGF stimulates laminin γ1 deposition which leads to integrin signaling and DLL4 induction; loss of α2 or α6 integrins mimics Dll4 silencing and induces excessive network branching.","method":"siRNA knockdown of integrins and DLL4, laminin adhesion assays, 3D sprouting angiogenesis assay, signaling pathway analysis","journal":"Circulation research","confidence":"Medium","confidence_rationale":"Tier 2 — siRNA knockdown with functional readout, pathway placed upstream of Notch","pmids":["21474814"],"is_preprint":false},{"year":2009,"finding":"DLL4 is inducible on dendritic cells by TLR activation (not by early inflammatory cytokines IL-1/IL-18); DLL4 on DCs promotes IL-17-producing T cell generation via upregulation of Rorc expression; both Rorc and Il17 gene promoters are direct transcriptional Notch targets.","method":"In vitro DC-T cell co-culture with TLR ligands; Notch inhibition; Rorc and Il17 promoter analysis; cytokine measurements","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 — promoter analysis with functional T cell differentiation readout, single lab","pmids":["19494260"],"is_preprint":false},{"year":2012,"finding":"KSHV vGPCR upregulates DLL4 through an ERK-dependent mechanism in lymphatic endothelial cells; DLL4-stimulated Notch4 signaling suppresses cell cycle genes in neighboring lymphatic endothelial cells, inducing cellular quiescence.","method":"KSHV gene expression studies, ERK inhibition, NF-κB inhibition, gene expression profiling, functional Notch signaling assay","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 — mechanistic pathway dissection using specific inhibitors and gene expression analysis","pmids":["19816565"],"is_preprint":false},{"year":2012,"finding":"LRF (leukemia/lymphoma related factor) acts as an erythroid-specific repressor of Dll4 expression; Lrf deletion in erythroblasts upregulates Dll4, sensitizing HSCs to T-cell instructive Notch signals in the bone marrow, leading to premature lymphoid differentiation and loss of HSC maintenance.","method":"In vivo mouse models with erythroblast-specific Lrf deletion; functional HSC assays; Dll4 expression analysis in erythroblasts","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 — cell-type-specific in vivo genetic model with defined HSC phenotype and clear pathway placement","pmids":["23134786"],"is_preprint":false},{"year":2012,"finding":"Dll4-Notch signaling between DN1 T cell progenitors and thymic DCs regulates thymic DC development and regulatory T cell homeostasis; pharmacological Dll4 blockade converts DN1 progenitors to immature DCs, which then expand Treg cells via a DC-dependent, MHC-II-dependent mechanism independent of Flt3.","method":"Anti-Dll4 antibody blockade; genetic inactivation models; thymectomy experiments; flow cytometry; DC-T cell co-culture","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 — multiple complementary genetic and pharmacological approaches with defined cellular phenotypes","pmids":["22547652"],"is_preprint":false},{"year":2015,"finding":"Cerebral cavernous malformation protein CCM1 controls DLL4-Notch3 signaling between endothelial cells and pericytes; CCM1 silencing in endothelial cells decreases DLL4 levels, reducing Notch3 activity in co-cultured pericytes; DLL4 stimulates Notch3 on brain pericytes, inducing expression of PDGFRB2, N-Cadherin, HBEGF, TGFB1, NG2, and S1P, enhancing pericyte adhesion and antiangiogenic function.","method":"siRNA knockdown of CCM1 in endothelial cells; EC-pericyte co-culture; Notch3 reporter assays; transgenic Ccm1/Ccm2 endothelial knockout mouse models","journal":"Stroke","confidence":"Medium","confidence_rationale":"Tier 2 — co-culture mechanistic studies with gene expression analysis and mouse models, single lab","pmids":["25791711"],"is_preprint":false},{"year":2016,"finding":"Dll4 fluctuations in individual endothelial cells drive sprout branching through heterogeneous phase patterns; pathologically high VEGF or DLL4 overexpression leads to Notch-dependent synchronization of Dll4 fluctuations within endothelial clusters, switching vessels from branching to expansion mode.","method":"Live imaging of Dll4 expression in mouse retina in vivo and embryonic stem cell-derived sprouting assays; DLL4 overexpression; Notch inhibition","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 — live imaging in vivo and in vitro with both gain and loss of function, multiple approaches","pmids":["27074663"],"is_preprint":false},{"year":2018,"finding":"The multiple PDZ domain protein MPDZ physically interacts with the intracellular C-terminus of DLL4 (and DLL1) and enables their interaction with the adherens junction protein Nectin-2; MPDZ inactivation leads to impaired DLL4-induced Notch signaling activity and increased blood vessel sprouting.","method":"Co-immunoprecipitation of MPDZ with DLL4 and Nectin-2; MPDZ gene inactivation in endothelial cells; Notch signaling reporter assays; embryonic mouse hindbrain vascular analysis","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1-2 — direct binding demonstrated by Co-IP, gene inactivation with defined vascular phenotype","pmids":["29620522"],"is_preprint":false},{"year":2019,"finding":"LPA4 and LPA6 receptors activate YAP/TAZ through Gα12/Gα13-Rho-ROCK signaling in endothelial cells; YAP/TAZ knockdown increases β-catenin- and NICD-mediated DLL4 expression; the LPA4/LPA6-Gα12/Gα13-YAP/TAZ axis thereby represses endothelial DLL4 expression to promote sprouting angiogenesis.","method":"Endothelial-specific Lpa4;Lpa6 double KO mice; siRNA knockdown of signaling components; fibrin gel sprouting assay; Notch inhibitor rescue","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — conditional double KO with in vivo rescue, multiple siRNA knockdowns, pathway epistasis","pmids":["31335323"],"is_preprint":false},{"year":2019,"finding":"High glucose activates a DLL4-Notch1 positive feedback loop in keratinocytes; Notch1 inactivation specifically in keratinocytes cancels the repressive effects of this loop on wound healing in diabetes, demonstrating that keratinocyte-specific Dll4-Notch1 signaling impairs diabetic wound healing.","method":"Loss-of-function genetic approaches (keratinocyte-specific Notch1 knockout); diabetic mouse wound healing models; pharmacological Notch inhibition","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — cell-type-specific genetic KO with defined physiological phenotype","pmids":["30886104"],"is_preprint":false},{"year":2019,"finding":"Indoxyl sulfate induces DLL4 protein upregulation in macrophages (partly through inhibition of the ubiquitin-proteasome pathway via the deubiquitinating enzyme USP5); DLL4 then activates Notch signaling to drive proinflammatory macrophage polarization; the uptake pathway is mediated by OATP2B1 transporter; macrophage-specific DLL4 knockout inhibits atherosclerosis in Ldlr-/- mice.","method":"Global proteomics; siRNA knockdown via macrophage-targeted lipid nanoparticles; macrophage-specific DLL4 KO; atherosclerosis mouse model; Dll4 antibody treatment","journal":"Circulation","confidence":"High","confidence_rationale":"Tier 2 — unbiased proteomics plus targeted genetic/pharmacological validation in vivo","pmids":["30586693"],"is_preprint":false},{"year":2020,"finding":"Dll4 acts as a negative regulator of intra-aortic hematopoietic cluster (IAHC) formation by impairing the recruitment of surrounding hemogenic cells into existing clusters; blocking Dll4 promotes entry of new hemogenic Gfi1+ cells into IAHCs and increases HSC numbers.","method":"Live imaging of organotypic slice cultures; clonal analysis; mathematical modeling; Dll4 blocking experiments","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — live imaging with clonal analysis and multiple validation approaches","pmids":["32149421"],"is_preprint":false},{"year":2022,"finding":"In skeletal muscle atrophy (disuse or diabetes), microvascular endothelium upregulates and releases DLL4, which activates muscular Notch2 without direct cell-cell contact; inhibition of the Dll4-Notch2 axis prevents muscle atrophy and promotes overloading-induced hypertrophy in mice.","method":"Mouse models of disuse and diabetic atrophy; Dll4-Notch2 axis inhibition; endothelial Dll4 overexpression; muscle mass/signaling analysis","journal":"Nature metabolism","confidence":"High","confidence_rationale":"Tier 2 — loss/gain of function in vivo with defined tissue phenotype, paracrine (non-contact) mechanism demonstrated","pmids":["35228746"],"is_preprint":false},{"year":2015,"finding":"DLL4 is expressed on a sub-population of bipotent hematoendothelial progenitors (HEPs) in hESCs; DLL4-high HEPs are enriched in endothelial potential while DLL4-low/-negative HEPs are committed to hematopoietic lineage; DLL4 stimulation enhances hematopoietic differentiation of HEPs and increases clonogenic hematopoietic progenitor output.","method":"hESC differentiation; clonal analysis; transcriptome profiling; confocal imaging of embryoid bodies; DLL4 stimulation assays","journal":"Leukemia","confidence":"Medium","confidence_rationale":"Tier 2 — clonal analysis with transcriptome; single lab","pmids":["25778099"],"is_preprint":false},{"year":2015,"finding":"DLL4 only is an efficient cis-inhibitor of Notch signaling whereas DLL1 has minimal cis-inhibitory activity; this differential cis-inhibition property contributes to functional divergence of DLL1 and DLL4 in tissue-specific contexts, explaining why transgenic DLL4 cannot replace DLL1 during somitogenesis.","method":"Conditional overexpression from Hprt locus; knock-in of Dll4 into Dll1 locus (Dll1Dll4ki); in vitro cis/trans Notch activation assays","journal":"PLoS genetics","confidence":"High","confidence_rationale":"Tier 1-2 — reconstitution of cis-inhibition in vitro combined with rigorous in vivo knock-in genetics","pmids":["26114479"],"is_preprint":false},{"year":2018,"finding":"TLR4 signaling in lung endothelial cells activates ERK phosphorylation, which causes ERK-dependent phosphorylation of FOXC2 and its transcriptional activation of the DLL4 gene; FOXC2-siRNA or ERK inhibition attenuates LPS-induced DLL4 expression and aberrant angiogenic sprouting both in vitro and in vivo.","method":"LPS stimulation of endothelial cells; pharmacological ERK inhibition; FOXC2 siRNA; ERK-2 dominant negative; FOXC2 ChIP at DLL4 promoter; neonatal mouse retinal angiogenesis model","journal":"The Journal of physiology","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP evidence for direct promoter binding plus in vivo confirmation; single lab","pmids":["29380370"],"is_preprint":false},{"year":2020,"finding":"RHOQ is induced by DLL4/Notch signaling and is essential for NICD nuclear translocation; in the absence of RHOQ, Notch1 is targeted for degradation in the autophagy pathway and NICD is sequestered from the nucleus and degraded in lysosomes, establishing a feed-forward mechanism.","method":"RHOQ siRNA knockdown; RHOQ overexpression; Notch signaling reporter assays; in vitro sprouting assay; zebrafish in vivo vascular analysis; subcellular localization studies","journal":"Angiogenesis","confidence":"Medium","confidence_rationale":"Tier 2 — mechanistic pathway placed with multiple cellular assays, in vivo validation; single lab","pmids":["32506201"],"is_preprint":false},{"year":2014,"finding":"Fibulin-3 activates ADAM10/17 in endothelial cells by inhibiting TIMP3, resulting in increased Notch cleavage and increased DLL4 expression independently of VEGF signaling; DLL4 knockdown reduces fibulin-3-dependent proangiogenic effects in vitro.","method":"ADAM10/17 inhibition; DLL4 siRNA knockdown; TIMP3 inhibition; endothelial cell motility and tubule formation assays; glioma xenograft models","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — mechanistic pathway dissection with siRNA and pharmacological inhibitors; single lab","pmids":["25139440"],"is_preprint":false},{"year":2019,"finding":"DLL4 in coronary arterial development functions within a Dll4-Jag1-EphrinB2 signaling cascade; Dll4 inactivation stimulates excessive capillary growth from sinus venosus, while forced Dll4 expression or Mfng overexpression blocks coronary plexus remodeling and arterial differentiation; EphrinB2 is a critical downstream effector of Dll4 in arterial morphogenesis.","method":"Endocardial-specific Jag1 and Dll4 conditional knockout mice; forced Dll4/Mfng expression; angiogenic rescue in ventricular explants and primary human ECs","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 — multiple conditional knockouts plus rescue experiments, pathway ordered","pmids":["31789590"],"is_preprint":false},{"year":2021,"finding":"In zebrafish valvulogenesis, blood flow-induced shear stress activates Notch signaling in endocardial cells via Dll4-mediated lateral inhibition, singling out Dll4-positive endocardial cells that ingress into the cardiac jelly in response to Wnt9a (produced through Erk5-Klf2-Wnt9a cascade); these parallel mechanosensitive pathways produce binary luminal/abluminal cell fate decisions.","method":"Zebrafish genetic models; live imaging; Notch/Wnt9a pathway manipulation; endocardial cell fate analysis","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo zebrafish imaging with genetic manipulation; single lab","pmids":["34610316"],"is_preprint":false},{"year":2021,"finding":"Pre-existing embryonic coronary plexus at the inner myocardium undergoes DLL4-NOTCH1 signaling-dependent angiogenic expansion to vascularize the expanding neonatal myocardium and to revascularize the regenerating neonatal heart.","method":"Lineage-tracing experiments; gain- and loss-of-function of Dll4-Notch1 in mice; live vascular imaging","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 — rigorous lineage tracing and gain/loss-of-function in vivo, highly informative study","pmids":["34497373"],"is_preprint":false},{"year":2019,"finding":"Soluble DLL4 activates Notch signaling in endothelial cells, increasing VE-cadherin at intercellular junctions and reducing vascular permeability through a cAMP/PKA-dependent pathway; PKA inhibition reverses DLL4-mediated barrier enhancement both in vitro and in vivo.","method":"Recombinant sDll4 treatment of EC monolayers; FITC-albumin permeability assay; γ-secretase inhibitor; PKA inhibition (H89); in vivo rat mesenteric microvessel hydraulic conductivity","journal":"American journal of physiology. Heart and circulatory physiology","confidence":"Medium","confidence_rationale":"Tier 1-2 — in vitro and in vivo functional assays with pharmacological mechanistic dissection; single lab","pmids":["30681366"],"is_preprint":false},{"year":2019,"finding":"TMZ treatment promotes nuclear translocation of MMP14 followed by extracellular release of DLL4; secreted DLL4 stimulates cleavage of Notch3, its nuclear translocation, and induction of GBM stemness and sphere formation.","method":"MMP14 expression and localization analysis after TMZ treatment in PDX GBM models; Kiloplex ELISA-based protein array; DLL4 functional and mechanistic studies; sphering capacity assays","journal":"International journal of cancer","confidence":"Medium","confidence_rationale":"Tier 2 — mechanistic pathway ordering with multiple assays; single lab","pmids":["31443114"],"is_preprint":false},{"year":2023,"finding":"Epsin1 modulates the sorting of DLL4 into tubular epithelial cell-derived exosomes under high-glucose conditions; exosomal DLL4 is captured by macrophages and promotes M1 macrophage activation via Notch1 (N1ICD) activation; Epsin1 knockdown in TECs reduces exosomal DLL4 and inhibits macrophage Notch1 activation.","method":"Mass spectrometry of urine exosomes; siRNA knockdown of Epsin1; TEC-macrophage co-culture with exosomes; in vivo mouse diabetic nephropathy model","journal":"Molecular therapy","confidence":"Medium","confidence_rationale":"Tier 2 — exosome-mediated delivery mechanism validated in vitro and in vivo; single lab","pmids":["37016580"],"is_preprint":false},{"year":2022,"finding":"DLL4 binds human and murine Notch receptors; affinity-matured DLL4 variant (DeltaMAX) has 500- to 1,000-fold increased receptor-binding affinity; DeltaMAX acts as agonist in plate/bead-bound format and as antagonist (soluble decoy) in reporter and neuronal differentiation assays, demonstrating dose/format-dependent agonist/antagonist activity.","method":"In vitro binding affinity assays; Notch reporter assays; neuronal differentiation assays; T cell stimulation assays; directed evolution/affinity maturation","journal":"Nature chemical biology","confidence":"High","confidence_rationale":"Tier 1 — biochemical reconstitution with mutagenesis and multiple functional assays","pmids":["36050494"],"is_preprint":false},{"year":2024,"finding":"Palmitic acid induces macrophage DLL4 expression, which triggers senescence in vascular smooth muscle cells (reducing collagen synthesis/deposition); macrophage-specific DLL4 knockout in atherosclerotic mice reduces plaque burden and improves plaque stability.","method":"Human cohort correlation; macrophage-specific DLL4 conditional KO in atherosclerotic mouse models; vascular smooth muscle cell senescence assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 — macrophage-specific conditional KO with defined plaque phenotype plus human data","pmids":["38346959"],"is_preprint":false},{"year":2015,"finding":"Heterozygous loss-of-function mutations (nonsense and missense, including cysteine-altering variants) in DLL4 cause autosomal-dominant Adams-Oliver syndrome, establishing DLL4 as an essential Notch ligand for vascular development in humans.","method":"Targeted resequencing of DLL4 in 89 AOS families; whole-exome/genome sequencing; candidate gene approach based on known DLL4 function","journal":"American journal of human genetics","confidence":"Medium","confidence_rationale":"Tier 3 — human genetics study; no in vitro functional validation of individual mutations reported","pmids":["26299364"],"is_preprint":false},{"year":2018,"finding":"DLL4 signaling via Notch1 impairs M2 macrophage differentiation and induces caspase3/7-dependent apoptosis selectively during M2 (but not M1) macrophage polarization; DLL4 upregulates pro-apoptotic effectors Bax, Bak, Bid, and Bim; fully differentiated M2 macrophages become resistant to DLL4 action.","method":"Human monocyte differentiation in vitro with immobilized recombinant DLL4; flow cytometry; qPCR; western blot for apoptotic pathway components; Notch inhibitors","journal":"Cell communication and signaling","confidence":"Medium","confidence_rationale":"Tier 2 — detailed mechanistic in vitro dissection with pathway inhibitors; single lab","pmids":["29321062"],"is_preprint":false},{"year":2016,"finding":"Endothelial DLL4 induces macrophage polarization into a proinflammatory M1 fate and elicits IL-6 production; both DLL4 and IL-6 are Notch-dependent and required for macrophage polarization; DLL4 upregulates M1-type markers and downregulates M2-type markers via Notch signaling in cardiac transplant rejection.","method":"EC/monocyte co-culture; endomyocardial biopsy analysis; flow cytometry; Notch signaling analysis","journal":"Biochemical pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 — co-culture mechanistic studies with clinical correlation; single lab","pmids":["26826491"],"is_preprint":false},{"year":2020,"finding":"Slug (SNAI2) suppresses Dll4-Notch signaling in endothelial cells to promote VEGFR2 expression; EC-specific Slug re-expression or loss of Dll4 rescues retinal angiogenesis in SlugKO mice; endothelial Slug is activated by SDF1α via CXCR4-ERK5 signaling.","method":"Slug endothelial-specific KO mice; Dll4 loss-of-function rescue; γ-secretase inhibition rescue; VEGF signaling inhibition; Notch target gene analysis","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 — genetic epistasis with multiple in vivo rescue experiments; pathway ordered","pmids":["33106502"],"is_preprint":false}],"current_model":"DLL4 is a transmembrane Notch ligand expressed predominantly in arterial and tip-cell endothelium (and also in other cell types including macrophages, thymic epithelium, and bone marrow mesenchymal cells) whose expression is induced by VEGF via PI3K/Akt and by hypoxia via HIF-1α, and which is transcriptionally activated combinatorially by Foxc1/2, SoxF factors, and Notch/RBPJ; DLL4 binding to Notch receptors (primarily Notch1, and also Notch2, Notch3, and Notch4 in non-endothelial contexts) suppresses tip cell formation and angiogenic sprouting through lateral inhibition while coupling sprouting to arterial specification, and is antagonized by Jagged1 (whose Notch-activating capacity is enhanced by Fringe glycosylation of Notch); its subcellular localization at adherens junctions is facilitated by MPDZ, downstream signaling is amplified by RHOQ-mediated NICD nuclear translocation, and it regulates diverse processes including intestinal stem cell maintenance, lymphatic vessel homeostasis, HSC niche signaling, T cell development, macrophage polarization, and skeletal muscle mass via the endothelial Dll4-muscular Notch2 axis."},"narrative":{"teleology":[{"year":2003,"claim":"Identifying how DLL4 expression is induced established it as a VEGF-responsive gene in arterial endothelium, linking growth factor signaling to Notch pathway activation.","evidence":"VEGF stimulation of arterial endothelial cells with PI3K/Akt and MAPK pathway inhibitors; 3D angiogenesis assays","pmids":["12482957"],"confidence":"High","gaps":["Whether other growth factors besides VEGF induce DLL4 in non-endothelial contexts","No structural basis for DLL4-Notch interaction at this point"]},{"year":2006,"claim":"Demonstration that DLL4 blockade causes endothelial hyperproliferation and that hypoxia/HIF-1α directly induces DLL4 established DLL4-Notch as a negative regulator of vascular growth and placed it downstream of both VEGF and oxygen sensing.","evidence":"Anti-DLL4 neutralizing antibody in tumor models; HIF-1α overexpression/knockdown in endothelial progenitors with promoter analysis","pmids":["17183323","17045587"],"confidence":"High","gaps":["Whether DLL4 acts in trans only or also in cis at this stage","Identity of downstream effectors beyond Hey1/Hey2"]},{"year":2007,"claim":"Showing that DLL4 is dynamically induced in retinal tip cells and limits sprouting via Notch established the lateral inhibition model in postnatal angiogenesis.","evidence":"DLL4-Fc and blocking antibody administration; Dll4 haploinsufficient mice; postnatal retinal vascular analysis","pmids":["17296940"],"confidence":"High","gaps":["How DLL4 oscillations relate to tip/stalk selection","Whether Notch receptor specificity matters in retinal endothelium"]},{"year":2008,"claim":"Identification of Foxc1/Foxc2 as direct transcriptional activators of the DLL4 promoter, with Foxc2 physically interacting with the Su(H)/NICD complex, revealed an integrative node linking VEGF-ERK/PI3K signaling to DLL4 transcription.","evidence":"Promoter reporter assays with Foxc binding element mutagenesis; co-immunoprecipitation of Foxc2 with NICD; siRNA knockdown in endothelial cells","pmids":["18545664"],"confidence":"High","gaps":["Whether Foxc factors are required in all vascular beds or only in specific territories","No ChIP-seq for genome-wide Foxc-DLL4 regulatory landscape"]},{"year":2009,"claim":"The discovery that Jagged1 antagonizes DLL4-Notch signaling through competition modulated by Fringe glycosyltransferases resolved how two Notch ligands produce opposing angiogenic outcomes in the same tissue.","evidence":"Genetic mouse models with Fringe manipulation; in vivo angiogenesis assays","pmids":["19524514"],"confidence":"High","gaps":["Quantitative binding affinities of glycosylated vs. unglycosylated Notch for DLL4 vs. Jagged1","Whether Fringe modulation operates identically in lymphatic endothelium"]},{"year":2010,"claim":"Wnt/β-catenin was added as a direct transcriptional inducer of DLL4, linking two major developmental signaling pathways in arteriovenous specification.","evidence":"Endothelial-specific β-catenin gain-of-function mice; Dll4 promoter chromatin/transcriptional analysis","pmids":["20627076"],"confidence":"High","gaps":["Whether β-catenin acts on the same enhancers as RBPJ/SoxF or distinct regulatory elements"]},{"year":2011,"claim":"Combinatorial knockout of Dll1 and Dll4 in intestinal epithelium revealed that these two ligands are the essential physiological Notch activators maintaining intestinal stem cells, expanding DLL4 function beyond vascular biology.","evidence":"Vil-Cre-ERT2 inducible single and double conditional knockouts in mice; stem cell lineage analysis","pmids":["21238454"],"confidence":"High","gaps":["Relative individual contributions of DLL1 vs. DLL4 to stem cell maintenance","Whether DLL4 acts on Lgr5+ stem cells directly or through Paneth cells"]},{"year":2011,"claim":"DLL4-Notch1 signaling was shown to regulate postnatal lymphatic sprouting via EphrinB2-dependent VEGFR3 signaling, and integrin-laminin adhesion was placed upstream of DLL4 induction, adding extracellular matrix cues to the regulatory network.","evidence":"Function-blocking antibodies against Notch1/Dll4 in lymphangiogenesis models; integrin siRNA knockdown with sprouting assays","pmids":["21700774","21474814"],"confidence":"High","gaps":["Whether integrin-mediated DLL4 induction operates in lymphatic endothelium","Molecular mechanism connecting integrin signaling to DLL4 transcription"]},{"year":2012,"claim":"Multiple discoveries extended DLL4-Notch to immune and hematopoietic biology: DLL4 was shown to regulate thymic DC/Treg homeostasis, post-transcriptionally controlled by miR-30, and its ectopic expression in erythroblasts (upon LRF loss) disrupted HSC maintenance.","evidence":"Anti-Dll4 blockade in thymic progenitors; miR-30 target-protector in zebrafish; erythroblast-specific Lrf KO mice with HSC assays","pmids":["22547652","23086751","23134786"],"confidence":"High","gaps":["Whether miR-30 regulation of DLL4 operates in non-endothelial cells","Mechanism by which erythroblast DLL4 reaches HSCs in the niche"]},{"year":2013,"claim":"Characterization of arterial-specific enhancers showed that SoxF factors and RBPJ/NICD combinatorially control DLL4 expression, with loss of both inputs ablating arterial identity, providing the cis-regulatory logic for arterial DLL4.","evidence":"Enhancer mutagenesis in transgenic mouse and zebrafish reporters; combined Sox7/Sox18/Rbpj knockdown","pmids":["23818617"],"confidence":"High","gaps":["Whether these enhancers operate in coronary or lymphatic arterial beds","Chromatin accessibility dynamics at these enhancers during de novo arteriogenesis"]},{"year":2015,"claim":"DLL4 was established in lymphatic vessel maintenance, bone marrow HSC niche signaling, coronary arterial development, and as unique among Delta ligands for potent cis-inhibition of Notch, while human genetics linked DLL4 haploinsufficiency to Adams-Oliver syndrome.","evidence":"Lymphatic EC-specific Dll4 KO with fat absorption assays; Ocn+ mesenchymal cell Dll4 deletion; Dll1Dll4ki knock-in; DLL4 mutation sequencing in AOS families","pmids":["26529256","25918341","26114479","26299364"],"confidence":"High","gaps":["Functional validation of individual AOS-associated DLL4 missense mutations","How DLL4 cis-inhibition is structurally distinct from DLL1","Whether released/soluble DLL4 contributes to niche signaling"]},{"year":2016,"claim":"Live imaging revealed that oscillatory DLL4 expression dynamics drive branching heterogeneity, with pathological VEGF or DLL4 levels synchronizing oscillations and switching from branching to expansion, adding a dynamic systems-level layer to DLL4 function.","evidence":"Live imaging of Dll4 in mouse retina and ES cell-derived sprouts; DLL4 overexpression; Notch inhibition","pmids":["27074663"],"confidence":"High","gaps":["Molecular clock or feedback controlling DLL4 oscillation period","Whether oscillation dynamics apply in non-retinal vascular beds"]},{"year":2018,"claim":"MPDZ was identified as a scaffold linking DLL4 to adherens junctions via Nectin-2, and ERK-dependent FOXC2 phosphorylation was shown to mediate TLR4-induced DLL4 transcription, refining understanding of DLL4 subcellular presentation and inflammatory induction.","evidence":"Co-immunoprecipitation of MPDZ-DLL4-Nectin-2; MPDZ KO vascular phenotype; FOXC2 ChIP at DLL4 promoter after LPS","pmids":["29620522","29380370"],"confidence":"High","gaps":["Whether MPDZ is required in all DLL4-expressing cell types","How MPDZ-mediated junctional localization affects signaling kinetics"]},{"year":2019,"claim":"Multiple studies revealed DLL4 operates in macrophage biology (proinflammatory polarization via indoxyl sulfate/USP5 stabilization, atherosclerosis), diabetic wound healing (keratinocyte DLL4-Notch1 loop), and coronary development (Dll4-Jag1-EphrinB2 cascade), while LPA4/6-YAP/TAZ was found to repress DLL4.","evidence":"Macrophage-specific DLL4 KO in atherosclerosis model; keratinocyte-specific Notch1 KO in diabetic wounds; endocardial Dll4/Jag1 conditional KOs; endothelial Lpa4/Lpa6 double KO mice","pmids":["30586693","30886104","31789590","31335323"],"confidence":"High","gaps":["Whether USP5-mediated DLL4 stabilization occurs in non-macrophage contexts","How YAP/TAZ mechanistically repress DLL4 transcription at the promoter level"]},{"year":2020,"claim":"RHOQ was identified as a Notch-induced GTPase required for NICD nuclear translocation, creating a feed-forward amplification loop; separately, DLL4 was shown to restrict intra-aortic hematopoietic cluster formation by limiting hemogenic cell recruitment.","evidence":"RHOQ siRNA/overexpression with subcellular localization and Notch reporter assays; live imaging of organotypic aortic slices with Dll4 blockade","pmids":["32506201","32149421"],"confidence":"Medium","gaps":["RHOQ mechanism validated primarily by siRNA in a single lab","Whether RHOQ functions downstream of DLL4-Notch specifically or all Notch ligands"]},{"year":2022,"claim":"A paracrine, non-contact-dependent endothelial DLL4–muscular Notch2 axis was discovered to regulate skeletal muscle mass, and affinity-matured DLL4 (DeltaMAX) demonstrated dose/format-dependent agonist-antagonist duality, revealing new signaling modalities.","evidence":"Mouse disuse/diabetic atrophy models with Dll4-Notch2 inhibition; directed evolution producing DeltaMAX with 500–1000-fold affinity increase; reporter and differentiation assays","pmids":["35228746","36050494"],"confidence":"High","gaps":["Whether released DLL4 signals in a soluble form or via exosomes in muscle","Structural basis for DeltaMAX agonist/antagonist switch"]},{"year":2024,"claim":"Macrophage DLL4 was shown to drive vascular smooth muscle cell senescence and plaque instability via palmitic acid induction, extending macrophage DLL4 from inflammation to atherosclerotic tissue remodeling.","evidence":"Macrophage-specific DLL4 conditional KO in atherosclerotic mice; VSMC senescence assays; human cohort correlation","pmids":["38346959"],"confidence":"High","gaps":["Whether DLL4-induced VSMC senescence is Notch1- or Notch3-dependent","Upstream lipid-sensing mechanism linking palmitic acid to DLL4 transcription"]},{"year":null,"claim":"Key unresolved questions include the structural basis of DLL4's unique cis-inhibitory potency versus DLL1, the precise mechanisms governing DLL4 oscillation dynamics, whether released/exosomal DLL4 represents a general paracrine signaling mode, and how DLL4 signals are decoded differently across its diverse tissue contexts.","evidence":"","pmids":[],"confidence":"Low","gaps":["No high-resolution structure of DLL4-Notch complex in membrane context","Mechanism of DLL4 oscillation periodicity unresolved","Exosomal vs. soluble DLL4 signaling capacity not systematically compared"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[1,2,39]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,29]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[22,39]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[38]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[27,36,37]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,2,3,5,22,39]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[5,8,33,35]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[12,16,19,25]},{"term_id":"R-HSA-1500931","term_label":"Cell-Cell communication","supporting_discovery_ids":[22,29]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[14,41]}],"complexes":[],"partners":["NOTCH1","NOTCH3","NOTCH4","MPDZ","JAG1","NECTIN2","FOXC2","RHOQ"],"other_free_text":[]},"mechanistic_narrative":"DLL4 is a transmembrane Notch ligand that functions as a central regulator of angiogenesis, vascular patterning, and cell fate specification across multiple tissues. In endothelial cells, VEGF induces DLL4 expression via PI3K/Akt signaling, and DLL4 activates Notch1 on neighboring cells through lateral inhibition to suppress tip cell formation and limit excessive sprouting; this negative feedback couples angiogenic sprouting to arterial specification, with Jagged1 serving as a competitive antagonist whose activity is modulated by Fringe glycosyltransferases [PMID:17296940, PMID:19524514, PMID:12482957, PMID:28714968]. DLL4 transcription is controlled combinatorially by Foxc1/2, SoxF factors, Notch/RBPJ autoregulatory inputs, Wnt/β-catenin, and HIF-1α, and its subcellular presentation at adherens junctions requires the scaffold protein MPDZ [PMID:18545664, PMID:23818617, PMID:20627076, PMID:17045587, PMID:29620522]. Beyond the vasculature, DLL4 together with DLL1 maintains intestinal stem cells, regulates lymphatic vessel integrity and fat absorption, drives thymus-seeding progenitor generation from bone marrow mesenchymal cells, directs macrophage proinflammatory polarization, and controls skeletal muscle mass via a paracrine endothelial DLL4–muscular Notch2 axis [PMID:21238454, PMID:26529256, PMID:25918341, PMID:30586693, PMID:35228746]. Heterozygous loss-of-function mutations in DLL4 cause autosomal-dominant Adams-Oliver syndrome [PMID:26299364]."},"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":"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":889,"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":790,"is_preprint":false},{"pmid":"17296940","id":"PMC_17296940","title":"Delta-like ligand 4 (Dll4) is induced by VEGF as a negative regulator of angiogenic sprouting.","date":"2007","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/17296940","citation_count":609,"is_preprint":false},{"pmid":"12482957","id":"PMC_12482957","title":"Regulation of Notch1 and Dll4 by vascular endothelial growth factor in arterial endothelial cells: implications for modulating arteriogenesis and angiogenesis.","date":"2003","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12482957","citation_count":414,"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":350,"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":324,"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":264,"is_preprint":false},{"pmid":"17692341","id":"PMC_17692341","title":"Regulation of multiple angiogenic pathways by Dll4 and Notch in human umbilical vein endothelial cells.","date":"2007","source":"Microvascular research","url":"https://pubmed.ncbi.nlm.nih.gov/17692341","citation_count":182,"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":177,"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":173,"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":"23086751","id":"PMC_23086751","title":"The microRNA-30 family targets DLL4 to modulate endothelial cell behavior during angiogenesis.","date":"2012","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/23086751","citation_count":151,"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":"18545664","id":"PMC_18545664","title":"Foxc transcription factors directly regulate Dll4 and Hey2 expression by interacting with the VEGF-Notch signaling pathways in endothelial cells.","date":"2008","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/18545664","citation_count":142,"is_preprint":false},{"pmid":"25918341","id":"PMC_25918341","title":"Specific bone cells produce DLL4 to generate thymus-seeding progenitors from bone marrow.","date":"2015","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/25918341","citation_count":128,"is_preprint":false},{"pmid":"19208840","id":"PMC_19208840","title":"Cross-talk between tumor and endothelial cells involving the Notch3-Dll4 interaction marks escape from tumor dormancy.","date":"2009","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/19208840","citation_count":125,"is_preprint":false},{"pmid":"23818617","id":"PMC_23818617","title":"Analysis of Dll4 regulation reveals a combinatorial role for Sox and Notch in arterial development.","date":"2013","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/23818617","citation_count":110,"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":106,"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":"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":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":86,"is_preprint":false},{"pmid":"29321062","id":"PMC_29321062","title":"Notch signaling triggered via the ligand DLL4 impedes M2 macrophage differentiation and promotes their apoptosis.","date":"2018","source":"Cell communication and signaling : CCS","url":"https://pubmed.ncbi.nlm.nih.gov/29321062","citation_count":82,"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":"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":75,"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":75,"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":74,"is_preprint":false},{"pmid":"30229512","id":"PMC_30229512","title":"A first-in-human phase 1a study of the bispecific anti-DLL4/anti-VEGF antibody navicixizumab (OMP-305B83) in patients with previously treated solid tumors.","date":"2018","source":"Investigational new drugs","url":"https://pubmed.ncbi.nlm.nih.gov/30229512","citation_count":65,"is_preprint":false},{"pmid":"21526177","id":"PMC_21526177","title":"Influence of Dll4 via HIF-1α-VEGF signaling on the angiogenesis of choroidal neovascularization under hypoxic conditions.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21526177","citation_count":64,"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":63,"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":"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":"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":53,"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":51,"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":"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":"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":"26826491","id":"PMC_26826491","title":"Notch signaling mediates crosstalk between endothelial cells and macrophages via Dll4 and IL6 in cardiac microvascular inflammation.","date":"2016","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/26826491","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":"25393540","id":"PMC_25393540","title":"Dll4 blockade potentiates the anti-tumor effects of VEGF inhibition in renal cell carcinoma patient-derived xenografts.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25393540","citation_count":46,"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":"23826258","id":"PMC_23826258","title":"Down-Regulated miR-30a in Clear Cell Renal Cell Carcinoma Correlated with Tumor Hematogenous Metastasis by Targeting Angiogenesis-Specific DLL4.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23826258","citation_count":44,"is_preprint":false},{"pmid":"23134786","id":"PMC_23134786","title":"LRF-mediated Dll4 repression in erythroblasts is necessary for hematopoietic stem cell maintenance.","date":"2012","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/23134786","citation_count":43,"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":"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":"25139440","id":"PMC_25139440","title":"Novel paracrine modulation of Notch-DLL4 signaling by fibulin-3 promotes angiogenesis in high-grade gliomas.","date":"2014","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/25139440","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":"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":"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":34,"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":"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":34,"is_preprint":false},{"pmid":"27049350","id":"PMC_27049350","title":"Simultaneous blockade of VEGF and Dll4 by HD105, a bispecific antibody, inhibits tumor progression and angiogenesis.","date":"2016","source":"mAbs","url":"https://pubmed.ncbi.nlm.nih.gov/27049350","citation_count":34,"is_preprint":false},{"pmid":"29592882","id":"PMC_29592882","title":"ABT-165, a Dual Variable Domain Immunoglobulin (DVD-Ig) Targeting DLL4 and VEGF, Demonstrates Superior Efficacy and Favorable Safety Profiles in Preclinical Models.","date":"2018","source":"Molecular cancer therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/29592882","citation_count":34,"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":32,"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":32,"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":31,"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":"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":"33383646","id":"PMC_33383646","title":"ABL001, a Bispecific Antibody Targeting VEGF and DLL4, with Chemotherapy, Synergistically Inhibits Tumor Progression in Xenograft Models.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33383646","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":"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":29,"is_preprint":false},{"pmid":"29674611","id":"PMC_29674611","title":"Inhibition of Dll4/Notch1 pathway promotes angiogenesis of Masquelet's induced membrane in rats.","date":"2018","source":"Experimental & molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29674611","citation_count":28,"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":28,"is_preprint":false},{"pmid":"26870802","id":"PMC_26870802","title":"Cyclic AMP Response Element Binding Protein Mediates Pathological Retinal Neovascularization via Modulating DLL4-NOTCH1 Signaling.","date":"2015","source":"EBioMedicine","url":"https://pubmed.ncbi.nlm.nih.gov/26870802","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":"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":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":"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":"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":"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":"32506201","id":"PMC_32506201","title":"RHOQ is induced by DLL4 and regulates angiogenesis by determining the intracellular route of the Notch intracellular domain.","date":"2020","source":"Angiogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/32506201","citation_count":23,"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":22,"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":"33249476","id":"PMC_33249476","title":"Nestin+/CD31+ cells in the hypoxic perivascular niche regulate glioblastoma chemoresistance by upregulating JAG1 and DLL4.","date":"2021","source":"Neuro-oncology","url":"https://pubmed.ncbi.nlm.nih.gov/33249476","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":"30063915","id":"PMC_30063915","title":"Matrine blocks AGEs- induced HCSMCs phenotypic conversion via suppressing Dll4-Notch pathway.","date":"2018","source":"European journal of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/30063915","citation_count":21,"is_preprint":false},{"pmid":"26472724","id":"PMC_26472724","title":"The vascular delta-like ligand-4 (DLL4)-Notch4 signaling correlates with angiogenesis in primary glioblastoma: an immunohistochemical study.","date":"2015","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/26472724","citation_count":21,"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":"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":"21517260","id":"PMC_21517260","title":"Correlation of Delta-like ligand 4 (DLL4) with VEGF and HIF-1α expression in human glioma.","date":"2011","source":"Asian Pacific journal of cancer prevention : APJCP","url":"https://pubmed.ncbi.nlm.nih.gov/21517260","citation_count":19,"is_preprint":false},{"pmid":"26828208","id":"PMC_26828208","title":"Activation of Dll4/Notch Signaling and Hypoxia-Inducible Factor-1 Alpha Facilitates Lymphangiogenesis in Lacrimal Glands in Dry Eye.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26828208","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":"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":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}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":51614,"output_tokens":10612,"usd":0.157011},"stage2":{"model":"claude-opus-4-6","input_tokens":14622,"output_tokens":4496,"usd":0.278265},"total_usd":0.435276,"stage1_batch_id":"msgbatch_0118RScg3WGaGkaZRtWZbtce","stage2_batch_id":"msgbatch_01Cpfzvtbj42aqp5oYZ5Te2s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2009,\n      \"finding\": \"Jagged1 antagonizes DLL4-Notch signaling in endothelial cells expressing Fringe family glycosyltransferases; glycosylation of Notch enhances DLL4-Notch signaling whereas Jagged1 has weak signaling capacity and competes with DLL4, establishing opposing roles for the two ligands in angiogenesis.\",\n      \"method\": \"Genetic mouse models, in vivo angiogenesis assays, functional studies with Fringe glycosyltransferases\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal genetic and biochemical approaches, highly cited foundational paper\",\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, demonstrating that DLL4-mediated Notch signaling regulates endothelial cell proliferation and differentiation and is crucial during active vascularization.\",\n      \"method\": \"Anti-DLL4 neutralizing antibody treatment, in vitro and in vivo endothelial cell assays, tumor models\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple in vitro and in vivo models, highly cited foundational paper\",\n      \"pmids\": [\"17183323\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"DLL4 expression is dynamically induced by VEGF in the retinal vasculature and acts as a negative feedback regulator to prevent overexuberant angiogenic sprouting; pharmacological inhibition of DLL4/Notch signaling (via soluble DLL4-Fc or blocking antibody) produces enhanced sprouting and increased endothelial proliferation.\",\n      \"method\": \"Intraocular administration of soluble DLL4-Fc and blocking antibody; Dll4 haploinsufficiency mouse model; postnatal retinal vascular analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic and pharmacological loss-of-function with defined vascular phenotypes, replicated by multiple approaches\",\n      \"pmids\": [\"17296940\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"VEGF, but not bFGF, induces DLL4 and Notch1 gene expression in human arterial endothelial cells via VEGFR1 and VEGFR2 through a PI3K/Akt pathway, independently of MAPK and Src; constitutive Notch activation stabilizes endothelial network formation.\",\n      \"method\": \"Gene expression analysis, pharmacological inhibitors of PI3K/Akt, MAPK, and Src; 3D angiogenesis model; Matrigel network formation assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple signaling pathway inhibitors used, orthogonal functional assays, highly cited\",\n      \"pmids\": [\"12482957\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Simultaneous genetic inactivation of Dll1 and Dll4 in mouse intestinal epithelium causes complete conversion of proliferating progenitors into postmitotic goblet cells with loss of intestinal stem cells, establishing that DLL1 and DLL4 together are the physiological Notch ligands required for intestinal stem cell maintenance.\",\n      \"method\": \"Inducible gut-specific gene targeting (Vil-Cre-ERT2) in mice; single and double conditional knockouts; lineage analysis with Notch1 reporter\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean genetic epistasis with specific stem cell phenotype, multiple mouse models\",\n      \"pmids\": [\"21238454\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Genetic experiments in postnatal mice reveal that the level of active Notch signaling (not direct DLL4-mediated cell-cell communication per se) is the key determinant of vessel growth; Notch activation directs tip-derived endothelial cells into developing arteries, coupling sprouting angiogenesis with artery formation. Endothelial VEGF-A and CXCR4 expression are key processes controlling Notch-dependent vessel growth.\",\n      \"method\": \"Endothelial-specific genetic targeting of Dll4 in tip cells in postnatal mice; conditional knockout models; retinal vascular analysis\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — rigorous in vivo genetic epistasis, multiple conditional mouse models\",\n      \"pmids\": [\"28714968\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Endothelial-specific stabilization of Wnt/β-catenin signaling upregulates Dll4 transcription and strongly increases Notch signaling in the endothelium, linking Wnt and Notch signaling pathways in vascular development and arteriovenous specification.\",\n      \"method\": \"Endothelial-specific β-catenin gain-of-function mouse models; in vitro β-catenin activation; chromatin/transcriptional analysis of Dll4 promoter\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — both in vivo and in vitro evidence with genetic models; mechanistic link demonstrated\",\n      \"pmids\": [\"20627076\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Foxc1 and Foxc2 transcription factors directly activate the Dll4 promoter and the Notch target Hey2 promoter via Foxc binding elements; Foxc2 physically interacts with a Notch transcriptional activation complex (Su(H)/NICD) to induce Hey2 promoter activity; VEGF-activated PI3K and ERK pathways modulate Foxc transcriptional activity in Dll4 and Hey2 induction.\",\n      \"method\": \"Promoter reporter assays, co-immunoprecipitation, siRNA knockdown, VEGF stimulation with PI3K/ERK inhibitors in endothelial cells\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct protein-protein interaction demonstrated by Co-IP; promoter functional validation by mutagenesis/binding\",\n      \"pmids\": [\"18545664\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Arterial Dll4 expression is regulated combinatorially by Notch signaling (via RBPJ/NICD direct binding) and SoxF transcription factors (Sox7, Sox18) through two arterial-specific enhancers; combinatorial loss of both SoxF and RBPJ binding ablates all Dll4 enhancer activity and results in loss of arterial markers and dorsal aorta.\",\n      \"method\": \"Arterial-specific enhancer characterization in mouse and zebrafish; transgenic reporter assays; combined Sox7/Sox18/Rbpj knockdown; endogenous dll4 expression analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — enhancer characterization with mutagenesis, genetic epistasis in two model organisms\",\n      \"pmids\": [\"23818617\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Hypoxia induces DLL4 expression through HIF-1α, which leads to activation of Notch target genes Hey1 and Hey2; in endothelial progenitor cells, hypoxia-mediated upregulation of DLL4 and Hey2 represses COUP-TFII (a venous identity regulator), promoting arterial cell fate; Hey factors create a negative feedback on HIF-1α-induced gene expression.\",\n      \"method\": \"Promoter analysis, HIF-1α overexpression/knockdown, endothelial progenitor cell culture under hypoxia, reporter assays\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — promoter analysis and functional cell-based assays, single lab\",\n      \"pmids\": [\"17045587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The microRNA-30 family directly targets the DLL4 3'UTR to repress DLL4 expression; miR-30b overexpression in endothelial cells increases vessel number/length in sprouting assays; microinjection of miR-30 mimics in zebrafish suppresses dll4 and causes excessive intersegmental vessel sprouting; target protector against the miR-30 site in dll4 3'UTR upregulates dll4.\",\n      \"method\": \"miRNA target site mutagenesis/target protector, overexpression in endothelial cells, zebrafish microinjection, sprouting angiogenesis assay\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — target protector validation in vivo and in vitro; multiple orthogonal methods\",\n      \"pmids\": [\"23086751\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DLL4 expression in intestinal lacteals requires VEGFR3 and VEGFR2 activation; genetic inactivation of Dll4 specifically in lymphatic endothelial cells leads to lacteal regression and impaired dietary fat uptake, establishing DLL4 as a regulator of adult lymphatic vessel maintenance and intestinal fat absorption.\",\n      \"method\": \"Lymphatic endothelial cell-specific Dll4 conditional knockout mice; VEGFR inhibition; dietary fat uptake assays; histological analysis\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — tissue-specific conditional knockout with defined functional phenotype\",\n      \"pmids\": [\"26529256\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DLL4 is expressed on bone marrow osteocalcin-positive (Ocn+) mesenchymal cells; selective depletion of DLL4 from these cells recapitulates thymopoietic abnormality (reduced thymus-seeding progenitors and T cell generation), establishing that bone marrow DLL4 drives thymus-seeding progenitor generation.\",\n      \"method\": \"In vivo deletion of DLL4 from Ocn+ cells; conditional cell-specific knockouts; progenitor frequency analysis; thymic function assays\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — cell-specific in vivo deletion with defined cellular phenotype\",\n      \"pmids\": [\"25918341\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Notch1-Dll4 signaling regulates postnatal lymphatic development; antibody blockade of Notch1 and Dll4 results in defective lymphatic sprouting associated with downregulation of EphrinB2 (which mediates VEGFR3/VEGFC signaling), dilation of collecting lymphatics with reduced mural cell coverage, and impaired wound-associated lymphangiogenesis.\",\n      \"method\": \"Function-blocking antibodies against Notch1 and Dll4 in mice; EphrinB2 expression analysis; wound healing lymphangiogenesis model\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological and pathway-level mechanistic analysis in vivo\",\n      \"pmids\": [\"21700774\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"DLL4 expression in endothelial cells of the tumor microenvironment activates Notch3 signaling in co-cultured T-ALL tumor cells, promoting their escape from dormancy; neutralization of DLL4 greatly reduces EC-mediated Notch3 activation in T-ALL cells and blocks tumorigenesis.\",\n      \"method\": \"EC-tumor cell co-culture, angiogenic factor stimulation, DLL4 neutralization, Notch3 silencing by RNAi, in vivo tumorigenicity assays\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-culture and RNAi mechanistic evidence, single lab\",\n      \"pmids\": [\"19208840\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Adhesion of endothelial cells to laminin-111 via α2β1 and α6β1 integrins triggers DLL4 expression and subsequent Notch pathway activation; VEGF stimulates laminin γ1 deposition which leads to integrin signaling and DLL4 induction; loss of α2 or α6 integrins mimics Dll4 silencing and induces excessive network branching.\",\n      \"method\": \"siRNA knockdown of integrins and DLL4, laminin adhesion assays, 3D sprouting angiogenesis assay, signaling pathway analysis\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — siRNA knockdown with functional readout, pathway placed upstream of Notch\",\n      \"pmids\": [\"21474814\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"DLL4 is inducible on dendritic cells by TLR activation (not by early inflammatory cytokines IL-1/IL-18); DLL4 on DCs promotes IL-17-producing T cell generation via upregulation of Rorc expression; both Rorc and Il17 gene promoters are direct transcriptional Notch targets.\",\n      \"method\": \"In vitro DC-T cell co-culture with TLR ligands; Notch inhibition; Rorc and Il17 promoter analysis; cytokine measurements\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — promoter analysis with functional T cell differentiation readout, single lab\",\n      \"pmids\": [\"19494260\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"KSHV vGPCR upregulates DLL4 through an ERK-dependent mechanism in lymphatic endothelial cells; DLL4-stimulated Notch4 signaling suppresses cell cycle genes in neighboring lymphatic endothelial cells, inducing cellular quiescence.\",\n      \"method\": \"KSHV gene expression studies, ERK inhibition, NF-κB inhibition, gene expression profiling, functional Notch signaling assay\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic pathway dissection using specific inhibitors and gene expression analysis\",\n      \"pmids\": [\"19816565\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"LRF (leukemia/lymphoma related factor) acts as an erythroid-specific repressor of Dll4 expression; Lrf deletion in erythroblasts upregulates Dll4, sensitizing HSCs to T-cell instructive Notch signals in the bone marrow, leading to premature lymphoid differentiation and loss of HSC maintenance.\",\n      \"method\": \"In vivo mouse models with erythroblast-specific Lrf deletion; functional HSC assays; Dll4 expression analysis in erythroblasts\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — cell-type-specific in vivo genetic model with defined HSC phenotype and clear pathway placement\",\n      \"pmids\": [\"23134786\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Dll4-Notch signaling between DN1 T cell progenitors and thymic DCs regulates thymic DC development and regulatory T cell homeostasis; pharmacological Dll4 blockade converts DN1 progenitors to immature DCs, which then expand Treg cells via a DC-dependent, MHC-II-dependent mechanism independent of Flt3.\",\n      \"method\": \"Anti-Dll4 antibody blockade; genetic inactivation models; thymectomy experiments; flow cytometry; DC-T cell co-culture\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple complementary genetic and pharmacological approaches with defined cellular phenotypes\",\n      \"pmids\": [\"22547652\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Cerebral cavernous malformation protein CCM1 controls DLL4-Notch3 signaling between endothelial cells and pericytes; CCM1 silencing in endothelial cells decreases DLL4 levels, reducing Notch3 activity in co-cultured pericytes; DLL4 stimulates Notch3 on brain pericytes, inducing expression of PDGFRB2, N-Cadherin, HBEGF, TGFB1, NG2, and S1P, enhancing pericyte adhesion and antiangiogenic function.\",\n      \"method\": \"siRNA knockdown of CCM1 in endothelial cells; EC-pericyte co-culture; Notch3 reporter assays; transgenic Ccm1/Ccm2 endothelial knockout mouse models\",\n      \"journal\": \"Stroke\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-culture mechanistic studies with gene expression analysis and mouse models, single lab\",\n      \"pmids\": [\"25791711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Dll4 fluctuations in individual endothelial cells drive sprout branching through heterogeneous phase patterns; pathologically high VEGF or DLL4 overexpression leads to Notch-dependent synchronization of Dll4 fluctuations within endothelial clusters, switching vessels from branching to expansion mode.\",\n      \"method\": \"Live imaging of Dll4 expression in mouse retina in vivo and embryonic stem cell-derived sprouting assays; DLL4 overexpression; Notch inhibition\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — live imaging in vivo and in vitro with both gain and loss of function, multiple approaches\",\n      \"pmids\": [\"27074663\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"The multiple PDZ domain protein MPDZ physically interacts with the intracellular C-terminus of DLL4 (and DLL1) and enables their interaction with the adherens junction protein Nectin-2; MPDZ inactivation leads to impaired DLL4-induced Notch signaling activity and increased blood vessel sprouting.\",\n      \"method\": \"Co-immunoprecipitation of MPDZ with DLL4 and Nectin-2; MPDZ gene inactivation in endothelial cells; Notch signaling reporter assays; embryonic mouse hindbrain vascular analysis\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct binding demonstrated by Co-IP, gene inactivation with defined vascular phenotype\",\n      \"pmids\": [\"29620522\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"LPA4 and LPA6 receptors activate YAP/TAZ through Gα12/Gα13-Rho-ROCK signaling in endothelial cells; YAP/TAZ knockdown increases β-catenin- and NICD-mediated DLL4 expression; the LPA4/LPA6-Gα12/Gα13-YAP/TAZ axis thereby represses endothelial DLL4 expression to promote sprouting angiogenesis.\",\n      \"method\": \"Endothelial-specific Lpa4;Lpa6 double KO mice; siRNA knockdown of signaling components; fibrin gel sprouting assay; Notch inhibitor rescue\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — conditional double KO with in vivo rescue, multiple siRNA knockdowns, pathway epistasis\",\n      \"pmids\": [\"31335323\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"High glucose activates a DLL4-Notch1 positive feedback loop in keratinocytes; Notch1 inactivation specifically in keratinocytes cancels the repressive effects of this loop on wound healing in diabetes, demonstrating that keratinocyte-specific Dll4-Notch1 signaling impairs diabetic wound healing.\",\n      \"method\": \"Loss-of-function genetic approaches (keratinocyte-specific Notch1 knockout); diabetic mouse wound healing models; pharmacological Notch inhibition\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — cell-type-specific genetic KO with defined physiological phenotype\",\n      \"pmids\": [\"30886104\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Indoxyl sulfate induces DLL4 protein upregulation in macrophages (partly through inhibition of the ubiquitin-proteasome pathway via the deubiquitinating enzyme USP5); DLL4 then activates Notch signaling to drive proinflammatory macrophage polarization; the uptake pathway is mediated by OATP2B1 transporter; macrophage-specific DLL4 knockout inhibits atherosclerosis in Ldlr-/- mice.\",\n      \"method\": \"Global proteomics; siRNA knockdown via macrophage-targeted lipid nanoparticles; macrophage-specific DLL4 KO; atherosclerosis mouse model; Dll4 antibody treatment\",\n      \"journal\": \"Circulation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — unbiased proteomics plus targeted genetic/pharmacological validation in vivo\",\n      \"pmids\": [\"30586693\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Dll4 acts as a negative regulator of intra-aortic hematopoietic cluster (IAHC) formation by impairing the recruitment of surrounding hemogenic cells into existing clusters; blocking Dll4 promotes entry of new hemogenic Gfi1+ cells into IAHCs and increases HSC numbers.\",\n      \"method\": \"Live imaging of organotypic slice cultures; clonal analysis; mathematical modeling; Dll4 blocking experiments\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — live imaging with clonal analysis and multiple validation approaches\",\n      \"pmids\": [\"32149421\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In skeletal muscle atrophy (disuse or diabetes), microvascular endothelium upregulates and releases DLL4, which activates muscular Notch2 without direct cell-cell contact; inhibition of the Dll4-Notch2 axis prevents muscle atrophy and promotes overloading-induced hypertrophy in mice.\",\n      \"method\": \"Mouse models of disuse and diabetic atrophy; Dll4-Notch2 axis inhibition; endothelial Dll4 overexpression; muscle mass/signaling analysis\",\n      \"journal\": \"Nature metabolism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — loss/gain of function in vivo with defined tissue phenotype, paracrine (non-contact) mechanism demonstrated\",\n      \"pmids\": [\"35228746\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DLL4 is expressed on a sub-population of bipotent hematoendothelial progenitors (HEPs) in hESCs; DLL4-high HEPs are enriched in endothelial potential while DLL4-low/-negative HEPs are committed to hematopoietic lineage; DLL4 stimulation enhances hematopoietic differentiation of HEPs and increases clonogenic hematopoietic progenitor output.\",\n      \"method\": \"hESC differentiation; clonal analysis; transcriptome profiling; confocal imaging of embryoid bodies; DLL4 stimulation assays\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clonal analysis with transcriptome; single lab\",\n      \"pmids\": [\"25778099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DLL4 only is an efficient cis-inhibitor of Notch signaling whereas DLL1 has minimal cis-inhibitory activity; this differential cis-inhibition property contributes to functional divergence of DLL1 and DLL4 in tissue-specific contexts, explaining why transgenic DLL4 cannot replace DLL1 during somitogenesis.\",\n      \"method\": \"Conditional overexpression from Hprt locus; knock-in of Dll4 into Dll1 locus (Dll1Dll4ki); in vitro cis/trans Notch activation assays\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — reconstitution of cis-inhibition in vitro combined with rigorous in vivo knock-in genetics\",\n      \"pmids\": [\"26114479\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"TLR4 signaling in lung endothelial cells activates ERK phosphorylation, which causes ERK-dependent phosphorylation of FOXC2 and its transcriptional activation of the DLL4 gene; FOXC2-siRNA or ERK inhibition attenuates LPS-induced DLL4 expression and aberrant angiogenic sprouting both in vitro and in vivo.\",\n      \"method\": \"LPS stimulation of endothelial cells; pharmacological ERK inhibition; FOXC2 siRNA; ERK-2 dominant negative; FOXC2 ChIP at DLL4 promoter; neonatal mouse retinal angiogenesis model\",\n      \"journal\": \"The Journal of physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP evidence for direct promoter binding plus in vivo confirmation; single lab\",\n      \"pmids\": [\"29380370\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RHOQ is induced by DLL4/Notch signaling and is essential for NICD nuclear translocation; in the absence of RHOQ, Notch1 is targeted for degradation in the autophagy pathway and NICD is sequestered from the nucleus and degraded in lysosomes, establishing a feed-forward mechanism.\",\n      \"method\": \"RHOQ siRNA knockdown; RHOQ overexpression; Notch signaling reporter assays; in vitro sprouting assay; zebrafish in vivo vascular analysis; subcellular localization studies\",\n      \"journal\": \"Angiogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic pathway placed with multiple cellular assays, in vivo validation; single lab\",\n      \"pmids\": [\"32506201\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Fibulin-3 activates ADAM10/17 in endothelial cells by inhibiting TIMP3, resulting in increased Notch cleavage and increased DLL4 expression independently of VEGF signaling; DLL4 knockdown reduces fibulin-3-dependent proangiogenic effects in vitro.\",\n      \"method\": \"ADAM10/17 inhibition; DLL4 siRNA knockdown; TIMP3 inhibition; endothelial cell motility and tubule formation assays; glioma xenograft models\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic pathway dissection with siRNA and pharmacological inhibitors; single lab\",\n      \"pmids\": [\"25139440\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"DLL4 in coronary arterial development functions within a Dll4-Jag1-EphrinB2 signaling cascade; Dll4 inactivation stimulates excessive capillary growth from sinus venosus, while forced Dll4 expression or Mfng overexpression blocks coronary plexus remodeling and arterial differentiation; EphrinB2 is a critical downstream effector of Dll4 in arterial morphogenesis.\",\n      \"method\": \"Endocardial-specific Jag1 and Dll4 conditional knockout mice; forced Dll4/Mfng expression; angiogenic rescue in ventricular explants and primary human ECs\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple conditional knockouts plus rescue experiments, pathway ordered\",\n      \"pmids\": [\"31789590\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In zebrafish valvulogenesis, blood flow-induced shear stress activates Notch signaling in endocardial cells via Dll4-mediated lateral inhibition, singling out Dll4-positive endocardial cells that ingress into the cardiac jelly in response to Wnt9a (produced through Erk5-Klf2-Wnt9a cascade); these parallel mechanosensitive pathways produce binary luminal/abluminal cell fate decisions.\",\n      \"method\": \"Zebrafish genetic models; live imaging; Notch/Wnt9a pathway manipulation; endocardial cell fate analysis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo zebrafish imaging with genetic manipulation; single lab\",\n      \"pmids\": [\"34610316\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Pre-existing embryonic coronary plexus at the inner myocardium undergoes DLL4-NOTCH1 signaling-dependent angiogenic expansion to vascularize the expanding neonatal myocardium and to revascularize the regenerating neonatal heart.\",\n      \"method\": \"Lineage-tracing experiments; gain- and loss-of-function of Dll4-Notch1 in mice; live vascular imaging\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — rigorous lineage tracing and gain/loss-of-function in vivo, highly informative study\",\n      \"pmids\": [\"34497373\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Soluble DLL4 activates Notch signaling in endothelial cells, increasing VE-cadherin at intercellular junctions and reducing vascular permeability through a cAMP/PKA-dependent pathway; PKA inhibition reverses DLL4-mediated barrier enhancement both in vitro and in vivo.\",\n      \"method\": \"Recombinant sDll4 treatment of EC monolayers; FITC-albumin permeability assay; γ-secretase inhibitor; PKA inhibition (H89); in vivo rat mesenteric microvessel hydraulic conductivity\",\n      \"journal\": \"American journal of physiology. Heart and circulatory physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro and in vivo functional assays with pharmacological mechanistic dissection; single lab\",\n      \"pmids\": [\"30681366\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TMZ treatment promotes nuclear translocation of MMP14 followed by extracellular release of DLL4; secreted DLL4 stimulates cleavage of Notch3, its nuclear translocation, and induction of GBM stemness and sphere formation.\",\n      \"method\": \"MMP14 expression and localization analysis after TMZ treatment in PDX GBM models; Kiloplex ELISA-based protein array; DLL4 functional and mechanistic studies; sphering capacity assays\",\n      \"journal\": \"International journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic pathway ordering with multiple assays; single lab\",\n      \"pmids\": [\"31443114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Epsin1 modulates the sorting of DLL4 into tubular epithelial cell-derived exosomes under high-glucose conditions; exosomal DLL4 is captured by macrophages and promotes M1 macrophage activation via Notch1 (N1ICD) activation; Epsin1 knockdown in TECs reduces exosomal DLL4 and inhibits macrophage Notch1 activation.\",\n      \"method\": \"Mass spectrometry of urine exosomes; siRNA knockdown of Epsin1; TEC-macrophage co-culture with exosomes; in vivo mouse diabetic nephropathy model\",\n      \"journal\": \"Molecular therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — exosome-mediated delivery mechanism validated in vitro and in vivo; single lab\",\n      \"pmids\": [\"37016580\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"DLL4 binds human and murine Notch receptors; affinity-matured DLL4 variant (DeltaMAX) has 500- to 1,000-fold increased receptor-binding affinity; DeltaMAX acts as agonist in plate/bead-bound format and as antagonist (soluble decoy) in reporter and neuronal differentiation assays, demonstrating dose/format-dependent agonist/antagonist activity.\",\n      \"method\": \"In vitro binding affinity assays; Notch reporter assays; neuronal differentiation assays; T cell stimulation assays; directed evolution/affinity maturation\",\n      \"journal\": \"Nature chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — biochemical reconstitution with mutagenesis and multiple functional assays\",\n      \"pmids\": [\"36050494\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Palmitic acid induces macrophage DLL4 expression, which triggers senescence in vascular smooth muscle cells (reducing collagen synthesis/deposition); macrophage-specific DLL4 knockout in atherosclerotic mice reduces plaque burden and improves plaque stability.\",\n      \"method\": \"Human cohort correlation; macrophage-specific DLL4 conditional KO in atherosclerotic mouse models; vascular smooth muscle cell senescence assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — macrophage-specific conditional KO with defined plaque phenotype plus human data\",\n      \"pmids\": [\"38346959\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Heterozygous loss-of-function mutations (nonsense and missense, including cysteine-altering variants) in DLL4 cause autosomal-dominant Adams-Oliver syndrome, establishing DLL4 as an essential Notch ligand for vascular development in humans.\",\n      \"method\": \"Targeted resequencing of DLL4 in 89 AOS families; whole-exome/genome sequencing; candidate gene approach based on known DLL4 function\",\n      \"journal\": \"American journal of human genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — human genetics study; no in vitro functional validation of individual mutations reported\",\n      \"pmids\": [\"26299364\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"DLL4 signaling via Notch1 impairs M2 macrophage differentiation and induces caspase3/7-dependent apoptosis selectively during M2 (but not M1) macrophage polarization; DLL4 upregulates pro-apoptotic effectors Bax, Bak, Bid, and Bim; fully differentiated M2 macrophages become resistant to DLL4 action.\",\n      \"method\": \"Human monocyte differentiation in vitro with immobilized recombinant DLL4; flow cytometry; qPCR; western blot for apoptotic pathway components; Notch inhibitors\",\n      \"journal\": \"Cell communication and signaling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — detailed mechanistic in vitro dissection with pathway inhibitors; single lab\",\n      \"pmids\": [\"29321062\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Endothelial DLL4 induces macrophage polarization into a proinflammatory M1 fate and elicits IL-6 production; both DLL4 and IL-6 are Notch-dependent and required for macrophage polarization; DLL4 upregulates M1-type markers and downregulates M2-type markers via Notch signaling in cardiac transplant rejection.\",\n      \"method\": \"EC/monocyte co-culture; endomyocardial biopsy analysis; flow cytometry; Notch signaling analysis\",\n      \"journal\": \"Biochemical pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-culture mechanistic studies with clinical correlation; single lab\",\n      \"pmids\": [\"26826491\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Slug (SNAI2) suppresses Dll4-Notch signaling in endothelial cells to promote VEGFR2 expression; EC-specific Slug re-expression or loss of Dll4 rescues retinal angiogenesis in SlugKO mice; endothelial Slug is activated by SDF1α via CXCR4-ERK5 signaling.\",\n      \"method\": \"Slug endothelial-specific KO mice; Dll4 loss-of-function rescue; γ-secretase inhibition rescue; VEGF signaling inhibition; Notch target gene analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis with multiple in vivo rescue experiments; pathway ordered\",\n      \"pmids\": [\"33106502\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DLL4 is a transmembrane Notch ligand expressed predominantly in arterial and tip-cell endothelium (and also in other cell types including macrophages, thymic epithelium, and bone marrow mesenchymal cells) whose expression is induced by VEGF via PI3K/Akt and by hypoxia via HIF-1α, and which is transcriptionally activated combinatorially by Foxc1/2, SoxF factors, and Notch/RBPJ; DLL4 binding to Notch receptors (primarily Notch1, and also Notch2, Notch3, and Notch4 in non-endothelial contexts) suppresses tip cell formation and angiogenic sprouting through lateral inhibition while coupling sprouting to arterial specification, and is antagonized by Jagged1 (whose Notch-activating capacity is enhanced by Fringe glycosylation of Notch); its subcellular localization at adherens junctions is facilitated by MPDZ, downstream signaling is amplified by RHOQ-mediated NICD nuclear translocation, and it regulates diverse processes including intestinal stem cell maintenance, lymphatic vessel homeostasis, HSC niche signaling, T cell development, macrophage polarization, and skeletal muscle mass via the endothelial Dll4-muscular Notch2 axis.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"DLL4 is a transmembrane Notch ligand that functions as a central regulator of angiogenesis, vascular patterning, and cell fate specification across multiple tissues. In endothelial cells, VEGF induces DLL4 expression via PI3K/Akt signaling, and DLL4 activates Notch1 on neighboring cells through lateral inhibition to suppress tip cell formation and limit excessive sprouting; this negative feedback couples angiogenic sprouting to arterial specification, with Jagged1 serving as a competitive antagonist whose activity is modulated by Fringe glycosyltransferases [PMID:17296940, PMID:19524514, PMID:12482957, PMID:28714968]. DLL4 transcription is controlled combinatorially by Foxc1/2, SoxF factors, Notch/RBPJ autoregulatory inputs, Wnt/β-catenin, and HIF-1α, and its subcellular presentation at adherens junctions requires the scaffold protein MPDZ [PMID:18545664, PMID:23818617, PMID:20627076, PMID:17045587, PMID:29620522]. Beyond the vasculature, DLL4 together with DLL1 maintains intestinal stem cells, regulates lymphatic vessel integrity and fat absorption, drives thymus-seeding progenitor generation from bone marrow mesenchymal cells, directs macrophage proinflammatory polarization, and controls skeletal muscle mass via a paracrine endothelial DLL4–muscular Notch2 axis [PMID:21238454, PMID:26529256, PMID:25918341, PMID:30586693, PMID:35228746]. Heterozygous loss-of-function mutations in DLL4 cause autosomal-dominant Adams-Oliver syndrome [PMID:26299364].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Identifying how DLL4 expression is induced established it as a VEGF-responsive gene in arterial endothelium, linking growth factor signaling to Notch pathway activation.\",\n      \"evidence\": \"VEGF stimulation of arterial endothelial cells with PI3K/Akt and MAPK pathway inhibitors; 3D angiogenesis assays\",\n      \"pmids\": [\"12482957\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other growth factors besides VEGF induce DLL4 in non-endothelial contexts\", \"No structural basis for DLL4-Notch interaction at this point\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Demonstration that DLL4 blockade causes endothelial hyperproliferation and that hypoxia/HIF-1α directly induces DLL4 established DLL4-Notch as a negative regulator of vascular growth and placed it downstream of both VEGF and oxygen sensing.\",\n      \"evidence\": \"Anti-DLL4 neutralizing antibody in tumor models; HIF-1α overexpression/knockdown in endothelial progenitors with promoter analysis\",\n      \"pmids\": [\"17183323\", \"17045587\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether DLL4 acts in trans only or also in cis at this stage\", \"Identity of downstream effectors beyond Hey1/Hey2\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Showing that DLL4 is dynamically induced in retinal tip cells and limits sprouting via Notch established the lateral inhibition model in postnatal angiogenesis.\",\n      \"evidence\": \"DLL4-Fc and blocking antibody administration; Dll4 haploinsufficient mice; postnatal retinal vascular analysis\",\n      \"pmids\": [\"17296940\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How DLL4 oscillations relate to tip/stalk selection\", \"Whether Notch receptor specificity matters in retinal endothelium\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identification of Foxc1/Foxc2 as direct transcriptional activators of the DLL4 promoter, with Foxc2 physically interacting with the Su(H)/NICD complex, revealed an integrative node linking VEGF-ERK/PI3K signaling to DLL4 transcription.\",\n      \"evidence\": \"Promoter reporter assays with Foxc binding element mutagenesis; co-immunoprecipitation of Foxc2 with NICD; siRNA knockdown in endothelial cells\",\n      \"pmids\": [\"18545664\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether Foxc factors are required in all vascular beds or only in specific territories\", \"No ChIP-seq for genome-wide Foxc-DLL4 regulatory landscape\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"The discovery that Jagged1 antagonizes DLL4-Notch signaling through competition modulated by Fringe glycosyltransferases resolved how two Notch ligands produce opposing angiogenic outcomes in the same tissue.\",\n      \"evidence\": \"Genetic mouse models with Fringe manipulation; in vivo angiogenesis assays\",\n      \"pmids\": [\"19524514\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quantitative binding affinities of glycosylated vs. unglycosylated Notch for DLL4 vs. Jagged1\", \"Whether Fringe modulation operates identically in lymphatic endothelium\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Wnt/β-catenin was added as a direct transcriptional inducer of DLL4, linking two major developmental signaling pathways in arteriovenous specification.\",\n      \"evidence\": \"Endothelial-specific β-catenin gain-of-function mice; Dll4 promoter chromatin/transcriptional analysis\",\n      \"pmids\": [\"20627076\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether β-catenin acts on the same enhancers as RBPJ/SoxF or distinct regulatory elements\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Combinatorial knockout of Dll1 and Dll4 in intestinal epithelium revealed that these two ligands are the essential physiological Notch activators maintaining intestinal stem cells, expanding DLL4 function beyond vascular biology.\",\n      \"evidence\": \"Vil-Cre-ERT2 inducible single and double conditional knockouts in mice; stem cell lineage analysis\",\n      \"pmids\": [\"21238454\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative individual contributions of DLL1 vs. DLL4 to stem cell maintenance\", \"Whether DLL4 acts on Lgr5+ stem cells directly or through Paneth cells\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"DLL4-Notch1 signaling was shown to regulate postnatal lymphatic sprouting via EphrinB2-dependent VEGFR3 signaling, and integrin-laminin adhesion was placed upstream of DLL4 induction, adding extracellular matrix cues to the regulatory network.\",\n      \"evidence\": \"Function-blocking antibodies against Notch1/Dll4 in lymphangiogenesis models; integrin siRNA knockdown with sprouting assays\",\n      \"pmids\": [\"21700774\", \"21474814\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether integrin-mediated DLL4 induction operates in lymphatic endothelium\", \"Molecular mechanism connecting integrin signaling to DLL4 transcription\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Multiple discoveries extended DLL4-Notch to immune and hematopoietic biology: DLL4 was shown to regulate thymic DC/Treg homeostasis, post-transcriptionally controlled by miR-30, and its ectopic expression in erythroblasts (upon LRF loss) disrupted HSC maintenance.\",\n      \"evidence\": \"Anti-Dll4 blockade in thymic progenitors; miR-30 target-protector in zebrafish; erythroblast-specific Lrf KO mice with HSC assays\",\n      \"pmids\": [\"22547652\", \"23086751\", \"23134786\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether miR-30 regulation of DLL4 operates in non-endothelial cells\", \"Mechanism by which erythroblast DLL4 reaches HSCs in the niche\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Characterization of arterial-specific enhancers showed that SoxF factors and RBPJ/NICD combinatorially control DLL4 expression, with loss of both inputs ablating arterial identity, providing the cis-regulatory logic for arterial DLL4.\",\n      \"evidence\": \"Enhancer mutagenesis in transgenic mouse and zebrafish reporters; combined Sox7/Sox18/Rbpj knockdown\",\n      \"pmids\": [\"23818617\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether these enhancers operate in coronary or lymphatic arterial beds\", \"Chromatin accessibility dynamics at these enhancers during de novo arteriogenesis\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"DLL4 was established in lymphatic vessel maintenance, bone marrow HSC niche signaling, coronary arterial development, and as unique among Delta ligands for potent cis-inhibition of Notch, while human genetics linked DLL4 haploinsufficiency to Adams-Oliver syndrome.\",\n      \"evidence\": \"Lymphatic EC-specific Dll4 KO with fat absorption assays; Ocn+ mesenchymal cell Dll4 deletion; Dll1Dll4ki knock-in; DLL4 mutation sequencing in AOS families\",\n      \"pmids\": [\"26529256\", \"25918341\", \"26114479\", \"26299364\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional validation of individual AOS-associated DLL4 missense mutations\", \"How DLL4 cis-inhibition is structurally distinct from DLL1\", \"Whether released/soluble DLL4 contributes to niche signaling\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Live imaging revealed that oscillatory DLL4 expression dynamics drive branching heterogeneity, with pathological VEGF or DLL4 levels synchronizing oscillations and switching from branching to expansion, adding a dynamic systems-level layer to DLL4 function.\",\n      \"evidence\": \"Live imaging of Dll4 in mouse retina and ES cell-derived sprouts; DLL4 overexpression; Notch inhibition\",\n      \"pmids\": [\"27074663\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular clock or feedback controlling DLL4 oscillation period\", \"Whether oscillation dynamics apply in non-retinal vascular beds\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"MPDZ was identified as a scaffold linking DLL4 to adherens junctions via Nectin-2, and ERK-dependent FOXC2 phosphorylation was shown to mediate TLR4-induced DLL4 transcription, refining understanding of DLL4 subcellular presentation and inflammatory induction.\",\n      \"evidence\": \"Co-immunoprecipitation of MPDZ-DLL4-Nectin-2; MPDZ KO vascular phenotype; FOXC2 ChIP at DLL4 promoter after LPS\",\n      \"pmids\": [\"29620522\", \"29380370\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether MPDZ is required in all DLL4-expressing cell types\", \"How MPDZ-mediated junctional localization affects signaling kinetics\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Multiple studies revealed DLL4 operates in macrophage biology (proinflammatory polarization via indoxyl sulfate/USP5 stabilization, atherosclerosis), diabetic wound healing (keratinocyte DLL4-Notch1 loop), and coronary development (Dll4-Jag1-EphrinB2 cascade), while LPA4/6-YAP/TAZ was found to repress DLL4.\",\n      \"evidence\": \"Macrophage-specific DLL4 KO in atherosclerosis model; keratinocyte-specific Notch1 KO in diabetic wounds; endocardial Dll4/Jag1 conditional KOs; endothelial Lpa4/Lpa6 double KO mice\",\n      \"pmids\": [\"30586693\", \"30886104\", \"31789590\", \"31335323\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether USP5-mediated DLL4 stabilization occurs in non-macrophage contexts\", \"How YAP/TAZ mechanistically repress DLL4 transcription at the promoter level\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"RHOQ was identified as a Notch-induced GTPase required for NICD nuclear translocation, creating a feed-forward amplification loop; separately, DLL4 was shown to restrict intra-aortic hematopoietic cluster formation by limiting hemogenic cell recruitment.\",\n      \"evidence\": \"RHOQ siRNA/overexpression with subcellular localization and Notch reporter assays; live imaging of organotypic aortic slices with Dll4 blockade\",\n      \"pmids\": [\"32506201\", \"32149421\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"RHOQ mechanism validated primarily by siRNA in a single lab\", \"Whether RHOQ functions downstream of DLL4-Notch specifically or all Notch ligands\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"A paracrine, non-contact-dependent endothelial DLL4–muscular Notch2 axis was discovered to regulate skeletal muscle mass, and affinity-matured DLL4 (DeltaMAX) demonstrated dose/format-dependent agonist-antagonist duality, revealing new signaling modalities.\",\n      \"evidence\": \"Mouse disuse/diabetic atrophy models with Dll4-Notch2 inhibition; directed evolution producing DeltaMAX with 500–1000-fold affinity increase; reporter and differentiation assays\",\n      \"pmids\": [\"35228746\", \"36050494\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether released DLL4 signals in a soluble form or via exosomes in muscle\", \"Structural basis for DeltaMAX agonist/antagonist switch\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Macrophage DLL4 was shown to drive vascular smooth muscle cell senescence and plaque instability via palmitic acid induction, extending macrophage DLL4 from inflammation to atherosclerotic tissue remodeling.\",\n      \"evidence\": \"Macrophage-specific DLL4 conditional KO in atherosclerotic mice; VSMC senescence assays; human cohort correlation\",\n      \"pmids\": [\"38346959\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether DLL4-induced VSMC senescence is Notch1- or Notch3-dependent\", \"Upstream lipid-sensing mechanism linking palmitic acid to DLL4 transcription\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis of DLL4's unique cis-inhibitory potency versus DLL1, the precise mechanisms governing DLL4 oscillation dynamics, whether released/exosomal DLL4 represents a general paracrine signaling mode, and how DLL4 signals are decoded differently across its diverse tissue contexts.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No high-resolution structure of DLL4-Notch complex in membrane context\", \"Mechanism of DLL4 oscillation periodicity unresolved\", \"Exosomal vs. soluble DLL4 signaling capacity not systematically compared\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [1, 2, 39]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 29]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [22, 39]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [38]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [27, 36, 37]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 2, 3, 5, 22, 39]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [5, 8, 33, 35]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [12, 16, 19, 25]},\n      {\"term_id\": \"R-HSA-1500931\", \"supporting_discovery_ids\": [22, 29]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [14, 41]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"NOTCH1\",\n      \"NOTCH3\",\n      \"NOTCH4\",\n      \"MPDZ\",\n      \"JAG1\",\n      \"NECTIN2\",\n      \"FOXC2\",\n      \"RHOQ\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}