Affinage

EPHB4

Ephrin type-B receptor 4 · UniProt P54760

Length
987 aa
Mass
108.3 kDa
Annotated
2026-04-28
100 papers in source corpus 32 papers cited in narrative 32 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

EphB4 is a transmembrane receptor tyrosine kinase that, through bidirectional signaling with its cognate ligand ephrin-B2 on adjacent cells, governs arteriovenous endothelial identity, vascular and lymphatic morphogenesis, bone homeostasis, hematopoietic progenitor mobilization, and tumor cell behavior. Forward signaling through the EphB4 kinase domain recruits RASA1 to suppress mTORC1 in endothelial cells, activates RhoA to reorganize the actin cytoskeleton, regulates collagen IV ER export required for angiogenesis, controls caveolae function and CLDN5-dependent lymphatic junctional integrity via Rac1/Rho, and in hepatocytes couples insulin receptor internalization through an AP2-clathrin pathway to lysosomal degradation, thereby modulating systemic insulin sensitivity (PMID:24837431, PMID:16950769, PMID:35015735, PMID:32897857, PMID:31782728, PMID:36131205). Loss-of-function mutations in EPHB4 cause capillary malformation–arteriovenous malformation type 2 (CM-AVM2) and primary venous valve aplasia in humans (PMID:28687708, PMID:34403370). In cancer, EphB4 exhibits context-dependent dual roles: ligand-independent kinase activity promotes survival via PI3K/AKT and migration via RhoA/FAK, whereas ephrin-B2-stimulated forward signaling triggers receptor internalization/degradation and can suppress tumor growth, consistent with its tumor-suppressor function in intestinal and head-and-neck carcinogenesis (PMID:22161689, PMID:19738063, PMID:35725568).

Mechanistic history

Synthesis pass · year-by-year structured walk · 16 steps
  1. 1994 High

    Establishing that EphB4 (HTK) possesses intrinsic tyrosine kinase activity and autophosphorylation capacity answered the foundational question of whether this orphan receptor signals through catalytic activity.

    Evidence In vitro kinase assay with purified protein and agonistic antibody stimulation in transfected NIH3T3 cells

    PMID:8188704

    Open questions at the time
    • Cognate ligand not yet identified
    • No downstream effectors known
    • No in vivo function established
  2. 1999 High

    Demonstrating that EphB4 knockout phenocopies ephrin-B2 knockout — with selective venous expression of EphB4 — established ephrin-B2 as the cognate ligand and proved that bidirectional ephrinB2/EphB4 signaling is required for angiogenesis and arteriovenous identity.

    Evidence Targeted gene knockout mice with comparative phenotypic analysis between EphB4-null and ephrin-B2-null embryos

    PMID:10518221

    Open questions at the time
    • Forward vs. reverse signaling contributions not individually resolved
    • Downstream intracellular effectors unknown
  3. 2003 Medium

    Dissecting forward versus reverse signaling showed they have non-overlapping functions: EphB4 forward signaling inhibits endothelial adhesion while reverse signaling through ephrin-B2 does not, clarifying how bidirectionality produces asymmetric cell behaviors at arteriovenous boundaries.

    Evidence Adhesion and migration assays with sorted primary endothelial cells using immobilized Fc-fusion proteins

    PMID:12588758

    Open questions at the time
    • Signaling intermediates downstream of forward signaling not identified
    • In vivo validation of adhesion phenotype lacking
  4. 2004 Medium

    Multiple studies established that EphB4 kinase activity is required for migration inhibition and that kinase-dead EphB4 can still drive angiogenesis in vivo through reverse ephrin-B2 signaling, resolving that the extracellular domain alone is sufficient for reverse but not forward signaling outputs.

    Evidence Kinase-dead and truncated EphB4 constructs in cell migration assays and xenograft models; pharmacological kinase inhibition

    PMID:14672701 PMID:15067119

    Open questions at the time
    • Specific kinase substrates not identified
    • Mechanism of reverse signaling transduction through ephrin-B2 not defined
  5. 2006 High

    A high-resolution crystal structure of the EphB4 ligand-binding domain revealed the hydrophobic cleft for ephrin-B2 engagement, providing the first atomic-level understanding of receptor-ligand specificity and enabling rational inhibitor design.

    Evidence X-ray crystallography at 1.65 Å with ITC validation of peptide binding determinants

    PMID:16472751

    Open questions at the time
    • Full ectodomain complex with ephrin-B2 not yet solved
    • No structure of the kinase domain
  6. 2006 High

    Three concurrent discoveries expanded EphB4's functional repertoire beyond vasculature: forward signaling through EphB4 enhances osteoblast differentiation while reverse signaling suppresses osteoclastogenesis via NFATc1; EphB4 promotes melanoma migration via RhoA-mediated actin reorganization; and EphB4 sustains breast cancer cell survival through PI3K/AKT, establishing kinase-dependent downstream effector pathways.

    Evidence Transgenic EphB4 overexpression in osteoblasts with bone histomorphometry; dominant-negative RhoA and kinase-dead constructs in melanoma cells; siRNA knockdown with caspase and AKT assays in breast cancer cells

    PMID:16816380 PMID:16890539 PMID:16950769

    Open questions at the time
    • Direct kinase substrates mediating RhoA activation unknown
    • Whether PI3K/AKT activation is direct or indirect unresolved
  7. 2008 High

    Genetic lineage tracing showed that ephrinB2/EphB4 sorts arterial and venous endothelial cells into correct vessels — a function mechanistically distinct from Notch-mediated control of arterial-venous proportions — clarifying the specific developmental role of this receptor pair.

    Evidence Mouse genetic loss-of-function with endothelial cell lineage tracing, compared with Notch gain/loss-of-function

    PMID:18952909

    Open questions at the time
    • Cell-autonomous sorting mechanism not molecularly defined
    • Whether sorting requires kinase activity or reverse signaling not resolved
  8. 2009 High

    Discovery that EphB4 inhibits integrin-mediated adhesion in a ligand-independent, kinase-dependent manner — reducing β1-integrin levels — revealed a constitutive signaling mode independent of ephrin-B2 engagement.

    Evidence siRNA, overexpression, and ephrin-binding-deficient point mutants with adhesion/spreading assays and β1-integrin western blots

    PMID:19552627

    Open questions at the time
    • Mechanism of β1-integrin downregulation not elucidated
    • Whether ligand-independent activity operates in normal physiology or only in overexpression contexts
  9. 2009 High

    EphB4 heterozygous inactivation in ApcMin mice accelerated intestinal tumorigenesis, establishing EphB4 as a haploinsufficient tumor suppressor in the intestine and showing its context-dependent oncogenic versus tumor-suppressive behavior.

    Evidence Genetic epistasis (ApcMin × EphB4+/−); tumor burden, invasion assays, xenograft models

    PMID:19738063

    Open questions at the time
    • Tumor-suppressive mechanism downstream of EphB4 not molecularly defined
    • Whether ligand-dependent activation is required for suppressor function unclear
  10. 2012 Medium

    Resolving the dual-role paradox: in the absence of ephrin-B2, overexpressed EphB4 promotes anchorage-independent growth; upon ligand stimulation, EphB4 is internalized and degraded via ERK1/2-dependent mechanisms, switching to tumor suppression — explaining how the same receptor can be oncogenic or suppressive depending on microenvironmental context.

    Evidence Ligand-stimulated vs. unstimulated EphB4 overexpression; receptor internalization, ERK1/2 phosphorylation, caspase-3/7 activity in prostate and mammary cell lines

    PMID:22161689

    Open questions at the time
    • Ubiquitin ligase or trafficking machinery for ligand-induced degradation not identified
    • Not validated in vivo
  11. 2014 High

    Identification of RASA1 as a direct effector recruited by EphB4 to suppress mTORC1 established the first complete signaling axis (EphB4→RASA1→mTORC1) for vascular development, and pharmacological mTORC1 inhibition rescued EphB4-deficient vascular defects, providing therapeutic proof-of-concept.

    Evidence Zebrafish genetics with engineered RASA1-binding-deficient EPHB4; mTORC1 activity assays; rapamycin rescue; patient AVM tissue phospho-S6 staining

    PMID:24837431

    Open questions at the time
    • Whether RASA1 acts via Ras-GAP activity or a GAP-independent mechanism on mTORC1 not fully resolved
    • Mammalian genetic validation of the full axis incomplete at this point
  12. 2017 High

    Discovery that loss-of-function EPHB4 mutations cause CM-AVM2 in humans — with in vitro confirmation of kinase loss — established the first Mendelian disease caused by EPHB4 deficiency and linked it to the RASA1-RAS-ERK pathway.

    Evidence Whole-exome sequencing in multiple families; in vitro kinase activity assays of patient missense mutations

    PMID:28687708

    Open questions at the time
    • Somatic second-hit mechanism not demonstrated
    • Genotype-phenotype correlations across mutation types not established
  13. 2019 High

    Inducible endothelial-specific EphB4 deletion in adults revealed organ-specific (cardiac) requirements for caveolae function, cell-cell adhesion under mechanical stress, and lipid transport, expanding EphB4's role from development to adult tissue homeostasis.

    Evidence Inducible EC-specific Ephb4 knockout mice; electron microscopy of caveolae; adhesion under flow; lipid transport assays

    PMID:31782728

    Open questions at the time
    • Why cardiac but not skeletal muscle capillaries are affected is unexplained
    • Direct molecular link between EphB4 and caveolae biogenesis unknown
  14. 2020 High

    EphrinB2/EphB4 signaling was shown to maintain lymphatic endothelial barrier function by controlling CLDN5 junctional localization through Rac1/Rho, establishing a mechanistic link to lymphatic-specific (not blood vascular) integrity.

    Evidence Conditional endothelial gene deletion in mice; CLDN5 immunofluorescence; Rac1/Rho activity assays in primary human lymphatic endothelial cells

    PMID:32897857

    Open questions at the time
    • How EphB4 selectively controls Rac1 vs. Rho balance in lymphatic but not blood endothelium not resolved
    • Direct phosphorylation targets linking EphB4 to CLDN5 trafficking unknown
  15. 2021 High

    Mutations in EPHB4 were shown to cause primary venous valve aplasia, with mechanistic dissection revealing that EphB4 controls valve-forming endothelial cell reorientation, polarity, and connexin37/43 junctional expression, expanding its disease spectrum beyond CM-AVM2.

    Evidence Human patient ultrasound; conditional Ephb4 and Efnb2 knockout mice; connexin37/43 immunostaining; cell polarity markers

    PMID:34403370

    Open questions at the time
    • Whether valve aplasia results from forward signaling, reverse signaling, or both is not resolved
    • Downstream pathway from EphB4 to connexin expression not defined
  16. 2022 High

    Four studies in 2022 greatly expanded the mechanistic scope: EphB4 directly binds the insulin receptor and promotes its AP2/clathrin-mediated endocytosis and lysosomal degradation in hepatocytes, regulating systemic insulin sensitivity; EphB4 kinase activity is required for collagen IV ER export independent of RASA1 binding; EphB4 forward signaling acts as a tumor suppressor in head-and-neck cancer; and EphB4/ephrinB2 promotes STAT3-driven LDLR transcription enhancing cholesterol uptake in liver metastases.

    Evidence Co-IP of EphB4/InsR with AP2 motif mutagenesis and in vivo glucose tolerance tests; EC-specific Ephb4 KO with RASA1-binding-deficient knock-in and pharmacological ER export rescue; genetically engineered mouse tumor models; STAT3/LDLR pathway analysis in metastasis models

    PMID:35015735 PMID:35725568 PMID:36131205 PMID:36376513

    Open questions at the time
    • Whether EphB4-InsR interaction requires ephrin-B2 or is ligand-independent in vivo
    • Kinase substrate mediating collagen IV ER export not identified
    • Mechanism by which EphB4 activates STAT3 for LDLR transcription not fully dissected

Open questions

Synthesis pass · forward-looking unresolved questions
  • Key unresolved questions include: the identity of direct EphB4 kinase substrates that mediate RhoA activation, caveolae regulation, and collagen IV ER export; the structural basis for EphB4's interaction with non-canonical ligands such as Epo and the insulin receptor; and how cell-type-specific signaling outputs (cardiac vs. lymphatic vs. hepatic) are determined.
  • No direct kinase substrate identified for most downstream phenotypes
  • Full-length EphB4 kinase domain structure not solved
  • Cell-type-specific signaling determinants remain undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 4 GO:0098772 molecular function regulator activity 2 GO:0060090 molecular adaptor activity 1
Localization
GO:0005886 plasma membrane 4 GO:0005783 endoplasmic reticulum 1
Pathway
R-HSA-162582 Signal Transduction 7 R-HSA-1643685 Disease 4 R-HSA-1266738 Developmental Biology 3 R-HSA-1430728 Metabolism 2 R-HSA-9612973 Autophagy 2 R-HSA-5653656 Vesicle-mediated transport 1
Partners
CLDN5EFNB2INSRPDGFRBRASA1RHOASDC1

Evidence

Reading pass · 32 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1994 EphB4 (originally cloned as HTK) is a transmembrane receptor tyrosine kinase of the EPH subfamily that autophosphorylates on tyrosine and exhibits intrinsic kinase activity in vitro; antibodies against its extracellular domain act as agonists inducing tyrosine phosphorylation in transfected cells. In vitro kinase assay with purified in vitro translated protein; immunoprecipitation; agonistic antibody stimulation in transfected NIH3T3 cells The Journal of biological chemistry High 8188704
1999 EphB4 is selectively expressed on venous (not arterial) endothelial cells in embryos, and targeted loss-of-function of EphB4 phenocopies the ephrin-B2 knockout, demonstrating that ephrin-B2/EphB4 interactions are intrinsically required in vascular endothelial cells for angiogenesis via bidirectional signaling. Targeted gene knockout in mice; comparative phenotypic analysis with ephrin-B2 null mice; immunohistochemistry for expression pattern Molecular cell High 10518221
2003 EphB4 forward signaling (via its kinase domain activated by ephrin-B2) inhibits endothelial cell adhesion, whereas EphB4-mediated reverse signaling through ephrin-B2 does not inhibit adhesion; cell migration is inhibited on immobilized ephrin-B2-Fc but not on EphB4-Fc, demonstrating distinct functional roles of forward vs. reverse signaling in arterial-venous endothelial cell interactions. Cell adhesion and migration assays using sorted primary endothelial cells and cell lines treated with immobilized Fc-fusion proteins Arteriosclerosis, thrombosis, and vascular biology Medium 12588758
2004 EphB4 on tumor cells promotes angiogenesis and tumor growth by interacting with ephrin-B2 on vascular endothelial cells; the extracellular domain of EphB4 attracts endothelial cells and stimulates endothelial cell invasion, survival, and proliferation in vitro, and EphB4 lacking the kinase domain (dominant negative) still promotes blood vessel enlargement in vivo via reverse ephrin-B2 signaling. Mouse xenograft model; in vitro endothelial cell migration/invasion/survival assays; EGFP-tagged kinase-dead EphB4 constructs Proceedings of the National Academy of Sciences of the United States of America Medium 15067119
2004 EphB4 signaling mediates ephrinB2-induced inhibition of cell migration; EphB4 kinase activity is essential for this effect, as demonstrated by relief of migration inhibition upon treatment with the Src-family kinase inhibitor PP2 (which also inhibits EphB4 kinase). Cell migration assay in a system solely dependent on ephrinB2-EphB4 signaling; pharmacological kinase inhibition with PP2 and PD153035 Biochemical and biophysical research communications Medium 14672701
2004 Upon EphB4 activation by ephrinB2, syndecan-1 and syntenin are upregulated in EphB4-positive endothelial cells in a dose- and time-dependent manner; upregulated syndecan-1 can compete with FGFR for bFGF, suppressing angiogenesis in vitro, but heparanase (present in inflamed tissue) cleaves syndecan-1 and converts the inhibitory effect to an activating one in vivo. cDNA microarray followed by RT-PCR; in vitro angiogenesis assay; preclustered ephrinB2 stimulation of endothelial cells Blood Medium 15126321
2006 Crystal structure of the EphB4 ligand-binding domain in complex with an antagonist peptide at 1.65 Å resolution reveals the peptide occupies the hydrophobic cleft corresponding to the ephrin-B2 G-H loop binding site; specific residues in this cleft determine EphB4 ligand specificity, and isothermal titration calorimetry identified peptide residues critical for receptor binding. X-ray crystallography at 1.65 Å; isothermal titration calorimetry with truncated peptide variants Structure (London, England : 1993) High 16472751
2006 Bidirectional ephrinB2-EphB4 signaling controls bone homeostasis: reverse signaling through ephrinB2 (expressed by osteoclast precursors) suppresses osteoclast differentiation by inhibiting the c-Fos–NFATc1 cascade, while forward signaling through EphB4 (expressed by osteoblasts) enhances osteoblast differentiation; transgenic overexpression of EphB4 in osteoblasts increases bone mass. Gain- and loss-of-function mouse experiments; transgenic EphB4 overexpression; analysis of c-Fos and NFATc1 signaling; bone histomorphometry Cell metabolism High 16890539
2006 EphB4 forward signaling promotes melanoma cell migration via activation of RhoA and consequent actin cytoskeleton reorganization; EphB4 kinase activity is required, as kinase-dead EphB4 inhibits migration and actin organization, and dominant negative RhoA blocks the pro-migratory effect of active EphB4. Overexpression/kinase-dead EphB4 constructs; dominant negative RhoA; cell migration assays; RhoA activity pull-down assay; actin cytoskeleton imaging The Journal of biological chemistry High 16950769
2006 EphB4 promotes breast cancer cell survival and acts as a survival factor; phosphorylation by its ligand EphrinB2 activates the PI3K/AKT pathway; EphB4 knockdown by siRNA induces apoptosis via caspase activation and sensitizes cells to TRAIL. siRNA knockdown; antisense oligonucleotides; cell viability and apoptosis assays; caspase activation assay; mouse xenograft model The American journal of pathology Medium 16816380
2008 EphrinB2/EphB4 signaling functions to sort arterial and venous endothelial cells into their respective vessels during angiogenesis; loss of ephrin B2 or EphB4 leads to mislocalization of venous-identity cells into the aorta, whereas Notch controls the relative size of arteries and veins by affecting arterial-to-venous endothelial cell proportions. Mouse genetic loss-of-function models; endothelial cell lineage tracing and localization analysis; comparison with Notch gain/loss-of-function Development (Cambridge, England) High 18952909
2009 EphB4 inhibits integrin-mediated cell-substrate adhesion, spreading, and migration in cancer cells in an ephrin-ligand-independent manner; this requires EphB4 kinase activity, and is associated with reduced β1-integrin protein levels. Mutations impairing ephrin binding do not affect this ligand-independent adhesion inhibition, demonstrating a constitutive, kinase-dependent function. siRNA knockdown; transient overexpression; single amino acid mutations impairing ephrin binding; cell adhesion/spreading/migration assays; western blotting for β1-integrin The Biochemical journal High 19552627
2009 EphB4 has tumor suppressor activities in intestinal tumorigenesis; inactivation of a single EphB4 allele in ApcMin mice results in increased proliferation, larger and more numerous intestinal tumors, and a 25% shorter lifespan; loss of EPHB4 in colon cancer cells increases invasive potential through extracellular matrix. Genetic mouse model (ApcMin × EphB4 heterozygous knockout); in vitro invasion assay; gene expression profiling; xenograft model Cancer research High 19738063
2010 EphB4 on tumor cells mediates site-specific metastatic dissemination by direct adhesive interaction with ephrin-B2 on endothelial cells; this was confirmed by atomic force microscopy at the single-cell level. The full intracellular domain (kinase activity) is required for organ-specific homing in vivo, as truncated EphB4 lacking the cytoplasmic domain does not promote metastatic dissemination. Atomic force microscopy (force spectroscopy); luciferase-based in vivo tumor cell trafficking; EphB4 truncation constructs; blocking with soluble EphB4-Fc Molecular cancer research : MCR High 21047731
2010 EphB4 forward kinase signaling is required for VEGF-driven angiogenesis; the specific small-molecule kinase inhibitor NVP-BHG712 inhibits EphB4 kinase activity at low nanomolar concentrations in cells, blocks EphB4 autophosphorylation in tissues after oral administration, and inhibits VEGF-induced vessel formation in vivo without directly affecting VEGFR kinase activity, demonstrating cross-talk between EphB4 and VEGFR signaling during angiogenesis. Biochemical kinase assay; cellular phosphorylation assay; kinase selectivity profiling; in vivo VEGF-driven angiogenesis model; pharmacokinetic analysis Angiogenesis High 20803239
2012 EphB4 has dual roles in cancer depending on ligand availability: overexpressed EphB4 in the absence of ephrin-B2 promotes anchorage-independent growth, migration, and invasion (ligand-independent tumor promotion); upon ephrin-B2 stimulation, EphB4 is internalized and degraded via a phospho-ERK1/2-dependent mechanism, reducing proliferation and increasing caspase-3/7 activity (ligand-dependent tumor suppression). Overexpression in prostate cancer and mammary epithelial cell lines; ephrin-B2-Fc stimulation; anchorage-independent growth assay; ERK1/2 phosphorylation western blot; caspase activity assay; receptor internalization assay International journal of cancer Medium 22161689
2013 EphB4 inhibition in esophageal cancer cells decreases phosphorylation of EphB4 and its downstream target p125FAK (focal adhesion kinase), and reduces cell migration, establishing FAK as a downstream effector of EphB4 forward signaling in cancer cells. Specific small-molecule EphB4 inhibitor; siRNA knockdown; cell migration assay; phosphotyrosine western blot; p125FAK immunoblot Cancer research Medium 23100466
2014 RASA1 (p120RasGAP) functions as a critical effector downstream of EPHB4 in endothelial cells to suppress mTORC1 activity; EPHB4 recruits RASA1 directly, and this interaction is required for normal blood vessel formation. Loss of EPHB4 or RASA1 in zebrafish leads to robustly over-activated mTORC1, and pharmacological mTORC1 inhibition rescues vascular defects in EPHB4-deficient animals, establishing the EPHB4/RASA1/mTORC1 signaling axis. Zebrafish genetic models; engineered EPHB4 receptors with altered RASA1 binding; rescue experiments; mTORC1 activity assay; pharmacological mTORC1 inhibition; patient AVM tissue analysis (phospho-S6 staining) The Journal of clinical investigation High 24837431
2014 PDGFRβ can activate EphB4 in a ligand-independent manner in rhabdomyosarcoma cells via PDGF ligand, converging on AKT and ERK1/2 signaling; conversely, EphB4 activation by its cognate ligand EphrinB2 does not activate PDGFRβ but instead paradoxically induces apoptosis, identifying EphB4 as a bivalent signaling node. Unbiased tyrosine kinome RNAi screen; co-immunoprecipitation; phospho-AKT/ERK western blot; apoptosis assay; dasatinib treatment in vitro and in vivo Proceedings of the National Academy of Sciences of the United States of America High 24733895
2015 EphB4 functions as an alternative erythropoietin (Epo) receptor that triggers downstream STAT3 signaling upon Epo binding, promoting tumor growth and progression independently of the canonical EpoR. Receptor identification experiments; STAT3 phosphorylation assay upon rhEpo stimulation; EphB4 knockdown/overexpression in cancer cell lines; in vivo tumor growth assays Cancer cell Medium 26481148
2016 EPHB4 regulates hematopoietic stem and progenitor cell (HSPC) mobilization from bone marrow through signaling interactions with ephrin B2 on hematopoietic cells; blockade of EPHB4/ephrin B2 signaling reduces HSPC mobilization and myeloid cell trafficking to tumors. Mutually exclusive localization of EPHB4 in sinusoids vs. ephrin B2 in hematopoietic cells by immunostaining; pharmacological blockade of EPHB4/ephrin B2 in mice; HSPC mobilization quantification; tumor infiltration assays The Journal of clinical investigation Medium 27820703
2017 Loss-of-function mutations in EPHB4 cause capillary malformation–arteriovenous malformation type 2 (CM-AVM2); in vitro expression of multiple patient missense mutations confirmed loss of EPHB4 kinase function; RASA1 (p120RasGAP), a direct effector of EPHB4, links this to deregulation of the RAS-ERK signaling pathway. Whole-exome sequencing; in vitro expression and kinase activity assay of mutant proteins; genetic linkage; patient cohort screening Circulation High 28687708
2018 A heterozygous EPHB4 splice-site mutation causing retention of a 12-bp intron reduces EPHB4 tyrosine phosphorylation (loss of function), leads to lymphatic vessel misbranching and developmental defects in zebrafish, and activates mTORC1 signaling; rescue with mTOR inhibitors or RAS-MAPK inhibitors restores normal vessel development. Whole exome sequencing; RNA-Seq splice analysis; transient co-expression of mutant/WT EPHB4 with phosphorylation assay; zebrafish morpholino; immunoblot for mTORC1 activity; pharmacological rescue; EPHB4 knock-in in HEK293T cells Human molecular genetics High 29905864
2019 Inducible, endothelial-cell-specific deletion of EphB4 in adult mice causes rupturing of cardiac capillaries, cardiomyocyte hypertrophy, and pathological cardiac remodeling; mechanistically, EphB4 controls caveolae function, cell-cell adhesion under mechanical stress, and lipid transport specifically in cardiac (but not skeletal muscle) capillaries. Inducible endothelial-specific Ephb4 knockout mice; cardiac histology and electron microscopy; caveolae functional analysis; cell-cell adhesion assay under flow; lipid transport assay; cultured endothelial cells eLife High 31782728
2019 EPHB4 inhibition in prostate cancer cells reduces GLUT3 expression, impairs glucose uptake and lowers cellular ATP, activating endoplasmic reticulum stress and immunogenic cell death (eIF2α phosphorylation, surface calreticulin, HMGB1/ATP release); this is linked to MYC downregulation via the SRC/p38 MAPK/4EBP1 signaling cascade. EPHB4 inhibition (small molecule and genetic); glucose uptake assay; ATP measurement; ER stress markers (eIF2α phosphorylation, calreticulin); HMGB1/ATP release assays; MYC western blot; SRC/p38/4EBP1 pathway analysis Cell death & disease Medium 31641103
2020 EphrinB2/EphB4 signaling controls lymphatic endothelial cell junction stability by regulating junctional localization of the tight junction protein CLDN5 and cytoskeletal contractility via Rac1/Rho signaling; conditional gene deletion in mice showed this is dispensable for blood but required for lymphatic endothelial barrier function. Conditional gene deletion in mice (endothelial-specific); primary human lymphatic endothelial cells; immunofluorescence for CLDN5 localization; Rac1/Rho activity assays; junctional integrity functional assay eLife High 32897857
2021 Mutations in EPHB4 cause primary venous valve aplasia in humans; conditional deletion of Ephb4 in mice disrupts valve-forming endothelial cell organization, reorientation, polarity, elongation, and proliferation, and loss of ephrinB2 disrupts junctional expression of connexin37 and connexin43 around reorienting valve-forming cells. Quantitative ultrasound in patients; conditional Ephb4 and Efnb2 knockout mice; GFP reporter for expression; immunostaining for connexin37/43, cell polarity, and proliferation markers JCI insight High 34403370
2022 EphB4 binds directly to the insulin receptor (InsR); this interaction is markedly enhanced by insulin and facilitates clathrin-mediated InsR endocytosis and lysosomal degradation via an AP2 complex-binding motif in EphB4; hepatic EphB4 overexpression decreases InsR levels and increases insulin resistance in mice, while EphB4 inhibition improves insulin resistance in obese mice. Co-immunoprecipitation of EphB4 and InsR; AP2 motif mutation analysis; clathrin-mediated endocytosis assay; lysosomal degradation assay; hepatic overexpression and genetic/pharmacological inhibition in mice; glucose tolerance and insulin resistance tests Nature metabolism High 36131205
2022 EPHB4 is required for export of collagen IV from the endoplasmic reticulum in vascular endothelial cells; loss of endothelial EPHB4 causes collagen IV accumulation in the ER, leading to EC apoptosis and defective angiogenesis; this function is independent of physical interaction with RASA1 but requires EPHB4 kinase activity, and can be rescued by drugs promoting collagen IV ER export or inhibiting Ras signaling. Induced EC-specific Ephb4 knockout mice; confocal imaging of collagen IV in ER; apoptosis assays; EPHB4 mutant knock-in (RASA1 binding-deficient); pharmacological rescue with ER export promoters and Ras pathway inhibitors JCI insight High 35015735
2022 EphB4 acts as a tumor suppressor in head and neck squamous cell carcinoma: loss of EphB4 intracellular domain on cancer cells accelerates tumor growth and angiogenesis with compensatory upregulation of EphA4 and increased regulatory T cell infiltration; ephrinB2 knockout on cancer cells and vasculature produces maximal tumor reduction and vascular normalization, establishing ephrinB2 as a tumor promoter and EphB4 as a tumor suppressor in this context. Genetically engineered mice; EphB4 and ephrinB2 knockout; recombinant constructs; pharmacologic agonists and antagonists; tumor immune microenvironment analysis; EphA4 expression analysis Nature communications High 35725568
2002 EphB4 signaling accelerates differentiation of human hematopoietic progenitor cells in a non-lineage-restricted manner, but preferentially promotes megakaryocytic and erythroid differentiation; effects are abrogated by mutations in select EphB4 tyrosine residues or by genistein (tyrosine kinase inhibitor), establishing tyrosine kinase activity and specific phosphorylation sites as required for these effects. Ectopic EphB4 expression in hematopoietic cell lines and cord blood CD34+ cells; site-directed mutagenesis of tyrosine residues; genistein inhibition; colony-forming and LTC-IC assays; differentiation marker expression Blood Medium 11929761
2022 EphB4 in colorectal cancer liver metastases interacts with EphrinB2 via forward signaling to enhance LDLR-mediated cholesterol uptake by promoting STAT3 phosphorylation which drives LDLR transcription; blocking the EFNB2/EphB4 axis reduces LDLR expression and cholesterol uptake in liver metastasis. In vitro and in vivo metastasis models; STAT3 phosphorylation assay; LDLR expression analysis; cholesterol uptake assay; LDLR rescue experiments Oncogene Medium 36376513

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2006 Bidirectional ephrinB2-EphB4 signaling controls bone homeostasis. Cell metabolism 596 16890539
1999 Symmetrical mutant phenotypes of the receptor EphB4 and its specific transmembrane ligand ephrin-B2 in cardiovascular development. Molecular cell 564 10518221
2004 Interplay between EphB4 on tumor cells and vascular ephrin-B2 regulates tumor growth. Proceedings of the National Academy of Sciences of the United States of America 211 15067119
2017 Germline Loss-of-Function Mutations in EPHB4 Cause a Second Form of Capillary Malformation-Arteriovenous Malformation (CM-AVM2) Deregulating RAS-MAPK Signaling. Circulation 205 28687708
2006 Receptor tyrosine kinase EphB4 is a survival factor in breast cancer. The American journal of pathology 165 16816380
2005 The soluble extracellular domain of EphB4 (sEphB4) antagonizes EphB4-EphrinB2 interaction, modulates angiogenesis, and inhibits tumor growth. Blood 136 16322467
2009 Preferential induction of EphB4 over EphB2 and its implication in colorectal cancer progression. Cancer research 120 19366806
2007 Paradoxes of the EphB4 receptor in cancer. Cancer research 113 17483308
2004 Inhibition of tumor growth and angiogenesis by soluble EphB4. Neoplasia (New York, N.Y.) 108 15153337
2010 The small molecule specific EphB4 kinase inhibitor NVP-BHG712 inhibits VEGF driven angiogenesis. Angiogenesis 104 20803239
2004 Expression of Ephb2 and Ephb4 in breast carcinoma. Pathology oncology research : POR 104 15029258
2008 Artery and vein size is balanced by Notch and ephrin B2/EphB4 during angiogenesis. Development (Cambridge, England) 98 18952909
2015 Erythropoietin Stimulates Tumor Growth via EphB4. Cancer cell 86 26481148
2006 The EphB4 receptor-tyrosine kinase promotes the migration of melanoma cells through Rho-mediated actin cytoskeleton reorganization. The Journal of biological chemistry 84 16950769
2009 The ephrinB2/EphB4 axis is dysregulated in osteoprogenitors from myeloma patients and its activation affects myeloma bone disease and tumor growth. Blood 82 19597185
2015 Erythropoietin promotes bone formation through EphrinB2/EphB4 signaling. Journal of dental research 79 25586589
2003 Distinct roles of ephrin-B2 forward and EphB4 reverse signaling in endothelial cells. Arteriosclerosis, thrombosis, and vascular biology 76 12588758
2006 Structure and thermodynamic characterization of the EphB4/Ephrin-B2 antagonist peptide complex reveals the determinants for receptor specificity. Structure (London, England : 1993) 75 16472751
1994 Cloning and characterization of HTK, a novel transmembrane tyrosine kinase of the EPH subfamily. The Journal of biological chemistry 74 8188704
2018 Pathogenic variant in EPHB4 results in central conducting lymphatic anomaly. Human molecular genetics 73 29905864
2010 Novel EphB4 monoclonal antibodies modulate angiogenesis and inhibit tumor growth. The American journal of pathology 71 20133814
2006 EphB4 provides survival advantage to squamous cell carcinoma of the head and neck. International journal of cancer 67 16615113
2014 RASA1 functions in EPHB4 signaling pathway to suppress endothelial mTORC1 activity. The Journal of clinical investigation 65 24837431
2013 The EphB4 receptor tyrosine kinase promotes lung cancer growth: a potential novel therapeutic target. PloS one 62 23844053
2002 A role of EphB4 receptor and its ligand, ephrin-B2, in erythropoiesis. Biochemical and biophysical research communications 60 12051776
2010 Communication between ephrinB2 and EphB4 within the osteoblast lineage. Advances in experimental medicine and biology 57 19950015
2009 The receptor tyrosine kinase EPHB4 has tumor suppressor activities in intestinal tumorigenesis. Cancer research 56 19738063
2012 Evidence for a dual function of EphB4 as tumor promoter and suppressor regulated by the absence or presence of the ephrin-B2 ligand. International journal of cancer 55 22161689
2008 Coexpression of EphB4 and ephrinB2 in tumour advancement of ovarian cancers. British journal of cancer 55 18231102
2017 Targeting receptor tyrosine kinase EphB4 in cancer therapy. Seminars in cancer biology 54 28993206
2020 Cantharidin treatment inhibits hepatocellular carcinoma development by regulating the JAK2/STAT3 and PI3K/Akt pathways in an EphB4-dependent manner. Pharmacological research 53 32407961
2009 Ephrin-independent regulation of cell substrate adhesion by the EphB4 receptor. The Biochemical journal 52 19552627
2016 EphrinB2/EphB4 pathway in postnatal angiogenesis: a potential therapeutic target for ischemic cardiovascular disease. Angiogenesis 48 27216867
2014 Ephrin B2/EphB4 mediates the actions of IGF-I signaling in regulating endochondral bone formation. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 48 24677183
2020 EphrinB2-EphB4 signalling provides Rho-mediated homeostatic control of lymphatic endothelial cell junction integrity. eLife 47 32897857
2019 EPHB4 inhibition activates ER stress to promote immunogenic cell death of prostate cancer cells. Cell death & disease 46 31641103
2019 Endothelial EphB4 maintains vascular integrity and transport function in adult heart. eLife 46 31782728
2005 Soluble forms of EphrinB2 and EphB4 reduce retinal neovascularization in a model of proliferative retinopathy. Investigative ophthalmology & visual science 45 15914639
2019 Inhibition of EphB4-Ephrin-B2 Signaling Reprograms the Tumor Immune Microenvironment in Head and Neck Cancers. Cancer research 44 30894369
2013 EphB4 enhances the process of endochondral ossification and inhibits remodeling during bone fracture repair. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 44 23165754
2012 Critical role for the receptor tyrosine kinase EPHB4 in esophageal cancers. Cancer research 44 23100466
2014 Ephrin B2 and EphB4 selectively mark arterial and venous vessels in cerebral arteriovenous malformation. The Journal of international medical research 41 24517927
2010 EphB4 promotes site-specific metastatic tumor cell dissemination by interacting with endothelial cell-expressed ephrinB2. Molecular cancer research : MCR 41 21047731
2006 Overexpression of ephrinB2 and EphB4 in tumor advancement of uterine endometrial cancers. Annals of oncology : official journal of the European Society for Medical Oncology 41 17108150
2005 Up-regulation of EphB4 in mesothelioma and its biological significance. Clinical cancer research : an official journal of the American Association for Cancer Research 40 15958611
2020 The critical role of the interplays of EphrinB2/EphB4 and VEGF in the induction of angiogenesis. Molecular biology reports 39 32488576
2002 Receptor tyrosine kinase, EphB4 (HTK), accelerates differentiation of select human hematopoietic cells. Blood 39 11929761
2016 EphB4 promotes the proliferation, invasion, and angiogenesis of human colorectal cancer. Experimental and molecular pathology 38 27072105
2020 CircRNA EPHB4 modulates stem properties and proliferation of gliomas via sponging miR-637 and up-regulating SOX10. Molecular oncology 37 33085838
2021 Sanguinarine combats hypoxia-induced activation of EphB4 and HIF-1α pathways in breast cancer. Phytomedicine : international journal of phytotherapy and phytopharmacology 36 33636580
2016 Sinusoidal ephrin receptor EPHB4 controls hematopoietic progenitor cell mobilization from bone marrow. The Journal of clinical investigation 35 27820703
2005 Soluble EphB4 regulates choroidal endothelial cell function and inhibits laser-induced choroidal neovascularization. Investigative ophthalmology & visual science 35 16303978
2019 Antitumor effects of circ-EPHB4 in hepatocellular carcinoma via inhibition of HIF-1α. Molecular carcinogenesis 34 30644610
2014 PDGFRβ reverses EphB4 signaling in alveolar rhabdomyosarcoma. Proceedings of the National Academy of Sciences of the United States of America 33 24733895
2018 Long noncoding RNA BC005927 upregulates EPHB4 and promotes gastric cancer metastasis under hypoxia. Cancer science 32 29383777
2010 Inhibitors of the tyrosine kinase EphB4. Part 3: identification of non-benzodioxole-based kinase inhibitors. Bioorganic & medicinal chemistry letters 32 20850301
2015 EphB4 Expressing Stromal Cells Exhibit an Enhanced Capacity for Hematopoietic Stem Cell Maintenance. Stem cells (Dayton, Ohio) 31 26033476
2010 Converging evidence for efficacy from parallel EphB4-targeted approaches in ovarian carcinoma. Molecular cancer therapeutics 31 20682653
2020 Homoharringtonine suppresses tumor proliferation and migration by regulating EphB4-mediated β-catenin loss in hepatocellular carcinoma. Cell death & disease 30 32801343
2004 Syndecan-1 up-regulated by ephrinB2/EphB4 plays dual roles in inflammatory angiogenesis. Blood 30 15126321
2018 Claudin 11 regulates bone homeostasis via bidirectional EphB4-EphrinB2 signaling. Experimental & molecular medicine 29 29700355
2016 KSR1 and EPHB4 Regulate Myc and PGC1β To Promote Survival of Human Colon Tumors. Molecular and cellular biology 28 27273865
2009 EPHB4 gene polymorphisms and risk of intracranial hemorrhage in patients with brain arteriovenous malformations. Circulation. Cardiovascular genetics 28 20031623
2022 Angiogenesis depends upon EPHB4-mediated export of collagen IV from vascular endothelial cells. JCI insight 27 35015735
2022 Insulin induces insulin receptor degradation in the liver through EphB4. Nature metabolism 27 36131205
2018 Inhibition of EphB4-Ephrin-B2 Signaling Enhances Response to Cetuximab-Radiation Therapy in Head and Neck Cancers. Clinical cancer research : an official journal of the American Association for Cancer Research 26 29848571
2009 Coexpression of EphB4 and ephrinB2 in tumor advancement of uterine cervical cancers. Gynecologic oncology 26 19356789
2020 Dysregulation of the EphrinB2-EphB4 ratio in pediatric cerebral arteriovenous malformations is associated with endothelial cell dysfunction in vitro and functions as a novel noninvasive biomarker in patients. Experimental & molecular medicine 25 32286515
2013 EphrinB2-EphB4 signals regulate formation and maintenance of funnel-shaped valves in corneal lymphatic capillaries. Investigative ophthalmology & visual science 25 23696610
2007 EphB4 expression along adult rat microvascular networks: EphB4 is more than a venous specific marker. Microcirculation (New York, N.Y. : 1994) 25 17454677
2009 Imidazo[1,2-a]pyrazine diaryl ureas: inhibitors of the receptor tyrosine kinase EphB4. Bioorganic & medicinal chemistry letters 24 19879134
2023 Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2/EphB4 signaling for orthodontic alveolar bone remodeling. Frontiers in bioengineering and biotechnology 23 37425367
2016 Low concentrations of TNF-α promote osteogenic differentiation via activation of the ephrinB2-EphB4 signalling pathway. Cell proliferation 23 27726217
2014 Arterial shear stress reduces eph-b4 expression in adult human veins. The Yale journal of biology and medicine 23 25191151
2013 Vascular endothelial growth factor-A inhibits EphB4 and stimulates delta-like ligand 4 expression in adult endothelial cells. The Journal of surgical research 23 23394931
2004 EphB4 signaling is capable of mediating ephrinB2-induced inhibition of cell migration. Biochemical and biophysical research communications 23 14672701
2022 Adaptive activation of EFNB2/EPHB4 axis promotes post-metastatic growth of colorectal cancer liver metastases by LDLR-mediated cholesterol uptake. Oncogene 22 36376513
2014 Compressive force regulates ephrinB2 and EphB4 in osteoblasts and osteoclasts contributing to alveolar bone resorption during experimental tooth movement. Korean journal of orthodontics 22 25473648
2010 Over-expression of Ephb4 is associated with carcinogenesis of gastric cancer. Digestive diseases and sciences 22 20686847
2015 Specific photothermal therapy to the tumors with high EphB4 receptor expression. Biomaterials 21 26264644
2012 Targeting the EphB4 receptor for cancer diagnosis and therapy monitoring. Molecular pharmaceutics 20 23211050
2012 Reduced adult endothelial cell EphB4 function promotes venous remodeling. American journal of physiology. Cell physiology 20 23269240
2022 EphrinB2-EphB4 Signaling in Neurooncological Disease. International journal of molecular sciences 19 35163601
2022 ED-71 inhibited osteoclastogenesis by enhancing EphrinB2-EphB4 signaling between osteoclasts and osteoblasts in osteoporosis. Cellular signalling 19 35690294
2022 EphB4 and ephrinB2 act in opposition in the head and neck tumor microenvironment. Nature communications 19 35725568
2018 EphB4: A promising target for upper aerodigestive malignancies. Biochimica et biophysica acta. Reviews on cancer 19 29369779
2017 EPHB4 is a therapeutic target in AML and promotes leukemia cell survival via AKT. Blood advances 19 29296810
2015 Anti-tumour effects of antibodies targeting the extracellular cysteine-rich region of the receptor tyrosine kinase EphB4. Oncotarget 19 25831049
2023 M6PR- and EphB4-Rich Exosomes Secreted by Serglycin-Overexpressing Esophageal Cancer Cells Promote Cancer Progression. International journal of biological sciences 18 36632458
2016 EPHB4 Regulates Human Trophoblast Cell Line HTR-8/SVneo Function: Implications for the Role of EPHB4 in Preeclampsia. Biology of reproduction 18 27512150
2014 Overexpression of EphB4, EphrinB2, and epidermal growth factor receptor in papillary thyroid carcinoma: A pilot study. Head & neck 18 24634162
2012 EphB4 is overexpressed in gliomas and promotes the growth of glioma cells. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 18 23138393
2021 Mutations in EPHB4 cause human venous valve aplasia. JCI insight 17 34403370
2021 Reconditioning of circulatory death hearts by ex-vivo machine perfusion with a novel HTK-N preservation solution. The Journal of heart and lung transplantation : the official publication of the International Society for Heart Transplantation 17 34420849
2020 Targeting the EphB4 receptor tyrosine kinase sensitizes HER2-positive breast cancer cells to Lapatinib. Cancer letters 17 32006616
2020 Icariin Alleviates Glucocorticoid-Induced Osteoporosis through EphB4/Ephrin-B2 Axis. Evidence-based complementary and alternative medicine : eCAM 17 32508946
2020 Ephrin-B2-EphB4 communication mediates tumor-endothelial cell interactions during hematogenous spread to spinal bone in a melanoma metastasis model. Oncogene 17 32989254
2018 Notch signaling promotes serrated neoplasia pathway in colorectal cancer through epigenetic modification of EPHB2 and EPHB4. Cancer management and research 17 30538561
2019 Pancreatic Tumor Microenvironment Modulation by EphB4-ephrinB2 Inhibition and Radiation Combination. Clinical cancer research : an official journal of the American Association for Cancer Research 16 30944125
2017 EphB4/EphrinB2 therapeutics in Rhabdomyosarcoma. PloS one 16 28817624