Affinage

NOTCH4

Neurogenic locus notch homolog protein 4 · UniProt Q99466

Length
2003 aa
Mass
209.6 kDa
Annotated
2026-04-29
100 papers in source corpus 38 papers cited in narrative 38 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NOTCH4 is an endothelial cell-enriched transmembrane receptor that governs vascular development, arteriovenous specification, and immune cell regulation through both canonical (RBP-Jκ/CBF1-dependent) and non-canonical signaling pathways. Upon proteolytic release, the Notch4 intracellular domain (ICD) translocates to the nucleus — a process negatively regulated by AKT-mediated phosphorylation and 14-3-3 binding (PMID:25740432) — where it activates transcription of targets including HEY1, SLUG, GAS1, and ephrinB2 via ankyrin repeat-dependent interactions with RBP-Jκ, while also inhibiting TGF-β signaling through direct Smad3 binding and suppressing NF-κB/TAK1 signaling by blocking TRAF6 autoubiquitination (PMID:16007227, PMID:29228365, PMID:32104513). Constitutive activation in endothelium causes arteriovenous malformations, enlarged vessels, and AV shunting that are fully reversible upon signal normalization through restoration of venous EphB4 expression (PMID:15994223, PMID:22261032), while in the immune system Notch4 on regulatory T cells subverts Treg function toward pro-inflammatory TH2/TH17 fates via Wnt/Hippo pathways and suppresses amphiregulin-mediated tissue repair (PMID:32929274, PMID:33915108). NOTCH4 transcription is controlled by AP-1, glucocorticoid receptor, PEA3/c-Jun, RUNX1, and YB-1, and in cancer the ICD drives stem cell maintenance, EMT, and therapy resistance, particularly in breast cancer where Notch4 activity is selectively enriched in tumor-initiating populations and JAG1-NOTCH4 signaling mediates anti-estrogen resistance (PMID:15684396, PMID:20068161, PMID:26387946).

Mechanistic history

Synthesis pass · year-by-year structured walk · 20 steps
  1. 1992 High

    Establishing that the intracellular domain of Notch4/int-3 is an autonomous oncogenic signal answered the foundational question of which portion of the protein drives transformation, showing that removal of the extracellular domain produces a constitutively active signaling moiety sufficient for mammary tumorigenesis.

    Evidence MMTV-driven transgenic mice expressing truncated int-3 ICD develop mammary adenocarcinomas; soft-agar transformation assay with ankyrin repeat-containing ICD in HC11 cells

    PMID:1312643 PMID:1372276

    Open questions at the time
    • Ligand and cleavage events that activate full-length Notch4 in physiological settings were unknown
    • Downstream transcriptional targets of the ICD were not identified
    • Whether oncogenic activity required RBP-Jκ interaction was untested
  2. 1996 High

    Demonstrating endothelial-specific expression of Notch4 during development established the primary physiological context, shifting focus from mammary oncogenesis to vascular biology.

    Evidence In situ hybridization across embryonic and adult mouse tissues showing endothelial-restricted expression

    PMID:8681805

    Open questions at the time
    • Functional requirement for Notch4 in endothelial biology was not yet tested
    • Whether Notch4 had non-redundant roles versus Notch1 in vasculature was unclear
  3. 1998 High

    Domain mapping revealed that the CBF-1 interaction domain plus ankyrin repeats constitute the minimal signaling unit for morphogenetic inhibition, and that Wnt-1 opposes Notch4, providing the first pathway epistasis data.

    Evidence Deletion constructs in TAC-2 mammary epithelial cells in 3D collagen gel branching morphogenesis assay with Wnt-1 co-expression

    PMID:9576833

    Open questions at the time
    • Whether ankyrin repeats act through RBP-Jκ-dependent or -independent routes was unresolved
    • Wnt-Notch crosstalk mechanism was not defined biochemically
  4. 2001 High

    Endothelial-specific constitutive Notch4 activation in vivo proved that Notch4 signaling is sufficient to disrupt vascular remodeling, causing embryonic lethality with dilated, disorganized vessels — the first direct in vivo evidence of Notch4's vascular function.

    Evidence Flk1 promoter-driven activated Notch4 knock-in transgenic mice with whole-mount and histological vascular analysis

    PMID:11344305

    Open questions at the time
    • Whether loss of Notch4 alone causes vascular defects was untested
    • Downstream effectors mediating vessel disorganization were unidentified
  5. 2002 High

    Identification of β1-integrin activation as a downstream effector explained how Notch4 inhibits angiogenic sprouting: by promoting integrin high-affinity conformation and increased adhesion rather than altering integrin expression.

    Evidence Endothelial sprouting assay, chick CAM assay, flow cytometry for integrin activation state

    PMID:11909975

    Open questions at the time
    • The signaling intermediates connecting Notch4-ICD to integrin activation were not identified
    • Contribution of RBP-Jκ-dependent versus -independent pathways to integrin effects was unclear
  6. 2003 High

    Dissection of anti-apoptotic signaling revealed that Notch4-ICD inhibits endothelial cell death through two parallel mechanisms — RBP-Jκ-dependent JNK inhibition and RBP-Jκ-independent Bcl-2 upregulation — establishing bifurcating downstream pathways.

    Evidence Notch4-ICD and deletion mutant overexpression with RBP-Jκ reporter assays and apoptosis measurement

    PMID:14701863

    Open questions at the time
    • Identity of the RBP-Jκ-independent pathway intermediates upstream of Bcl-2 was unknown
    • Relevance of anti-apoptotic function to in vivo vascular phenotypes was not demonstrated
  7. 2004 High

    Systematic domain deletion refined the sprouting inhibition mechanism to the ankyrin repeats as the minimal sufficient domain, operating through both RBP-Jκ-dependent and -independent routes.

    Evidence Quantitative endothelial sprouting assay with ankyrin repeat, RAM domain, and C-terminal deletion mutants plus RBP-Jκ reporters

    PMID:15187023

    Open questions at the time
    • Direct binding partners of the ankyrin repeats mediating RBP-Jκ-independent effects were not identified
  8. 2005 High

    Three parallel advances resolved transcriptional regulation, TGF-β crosstalk, and in vivo AVM pathobiology: AP-1/GR control NOTCH4 endothelial transcription; Notch4-ICD binds Smad3 to block TGF-β signaling; and constitutive Notch4 in adult endothelium causes reversible arteriovenous malformations with venous arterialization.

    Evidence ChIP and promoter mutagenesis for AP-1/GR; co-IP and domain mapping for Smad3 binding; tetracycline-repressible endothelial transgenic system with histology and ephrinB2 expression

    PMID:15684396 PMID:15994223 PMID:16007227

    Open questions at the time
    • How Smad3 binding is coordinated with RBP-Jκ interaction was unresolved
    • Upstream signals triggering Notch4 activation in AVM context were unknown
    • Whether AP-1-mediated transcription is required for AVM was not tested
  9. 2008 High

    Genetic epistasis using conditional Rbpj knockout demonstrated that Notch4-driven mammary developmental arrest requires RBP-Jκ, but Notch4-driven mammary tumorigenesis does not, definitively separating RBP-Jκ-dependent developmental from RBP-Jκ-independent oncogenic signaling. Simultaneously, brain-specific constitutive Notch4 was shown to cause cerebral AVMs with hemorrhage and neuronal death, reversible upon signal normalization.

    Evidence Conditional Rbpj knockout with Wap-Cre in mammary glands; tetracycline-repressible endothelial transgenic system in brain with MRI and behavioral assays

    PMID:18667694 PMID:18836481

    Open questions at the time
    • The RBP-Jκ-independent effectors driving mammary tumorigenesis were unidentified
    • Whether human cerebral AVMs involve NOTCH4 mutations was not established
  10. 2010 High

    Discovery that Notch4 signaling is selectively enriched 8-fold in breast cancer stem cells (versus Notch1 being lower) and that Notch4 inhibition abolishes tumor initiation established Notch4 as the principal Notch receptor maintaining cancer stem cell activity.

    Evidence Stem cell enrichment, Notch reporter assays, GSI and siRNA inhibition, in vivo tumor formation in breast cancer models

    PMID:20068161

    Open questions at the time
    • Specific Notch4 target genes in cancer stem cells were not identified at this point
    • Whether Notch4 stem cell function is RBP-Jκ-dependent was untested
  11. 2012 High

    Live imaging revealed that AVM regression upon Notch4 normalization occurs through vessel narrowing (not EC loss) and requires restoration of venous EphB4 expression, identifying EphB4 as a critical downstream mediator of AV identity.

    Evidence 4D two-photon imaging through cranial windows in Notch4* mice with EphB4 expression analysis and blood flow measurement

    PMID:22261032

    Open questions at the time
    • Whether EphB4 restoration is sufficient for AVM regression was not tested
    • Transcriptional mechanism linking Notch4 to EphB4 repression was not defined
  12. 2014 High

    A surprising finding that full-length NOTCH4 does not signal canonically in response to ligands but instead inhibits NOTCH1 in cis via its extracellular domain reframed Notch4 as both a signaling receptor (when cleaved) and a cis-inhibitor of Notch1, with Notch4-null mice showing only mild retinal vascular delay.

    Evidence Notch signaling reporter assays in cultured cells, full coding region Notch4 knockout mouse, cis-inhibition constructs, retinal vascular imaging

    PMID:24667410

    Open questions at the time
    • In vivo significance of cis-inhibition in specific vascular beds was not established
    • Whether Notch4 inhibits other Notch paralogs beyond Notch1 was untested
    • Structural basis for cis-inhibition was not determined
  13. 2015 High

    AKT-mediated phosphorylation of Notch4-ICD at four sites creates 14-3-3 binding sites that sequester the ICD in the cytoplasm, establishing PI3K/AKT as a negative regulator of Notch4 nuclear signaling and linking growth factor signaling to Notch4 output.

    Evidence In vitro kinase assay, site-directed mutagenesis, co-IP with 14-3-3, PI3K inhibitor treatment, subcellular fractionation

    PMID:25740432

    Open questions at the time
    • Whether AKT-mediated regulation operates in endothelial cells in vivo was not shown
    • The phosphatase(s) reversing this modification were not identified
  14. 2015 High

    JAG1 was identified as the activating ligand for NOTCH4 in anti-estrogen-treated breast cancer, and JAG1-NOTCH4 targeting reversed therapy-induced cancer stem cell expansion in patient-derived xenografts, establishing the ligand-receptor pair driving endocrine resistance.

    Evidence Patient-derived samples, PDX models, NOTCH4 targeting, ALDH1/mammosphere assays

    PMID:26387946

    Open questions at the time
    • Whether JAG1-NOTCH4 signaling is ligand-cleavage-dependent or involves the cis-inhibition mode was not distinguished
    • Downstream transcriptional program in resistant cancer stem cells was incompletely defined
  15. 2016 High

    Direct target gene identification showed Notch4-ICD activates Hey1/Hey2 transcription factors that bind and repress Snail2 and Twist1 promoters, demonstrating Notch4 can suppress invasion in melanoma — context-dependent signaling opposite to its pro-EMT role in other cancers.

    Evidence EMSA and luciferase reporter assays for Hey1/Hey2 binding to Snail2/Twist1 promoters, N4ICD overexpression with invasion/migration assays

    PMID:26801977

    Open questions at the time
    • Why Notch4 promotes EMT in breast cancer but suppresses invasion in melanoma was mechanistically unexplained
    • Whether chromatin context differs between these cell types was not addressed
  16. 2017 High

    Identification of RUNX1 as a negative transcriptional regulator of NOTCH4, with NOTCH4 knockout enhancing megakaryopoiesis, expanded Notch4's role beyond endothelium to hematopoietic lineage decisions.

    Evidence ChIP for RUNX1 at NOTCH4 locus, CRISPR-Cas9 NOTCH4 knockout in iPSCs, megakaryocyte differentiation assays

    PMID:29101237

    Open questions at the time
    • Whether Notch4 affects megakaryocyte function beyond differentiation was unknown
    • The specific Notch4-dependent transcriptional program in megakaryocyte progenitors was not characterized
  17. 2018 High

    Notch4-ICD was shown to physically interact with TAK1 and block TRAF6 autoubiquitination, revealing a non-transcriptional cytoplasmic mechanism by which Notch4 suppresses innate immune activation during mycobacterial infection.

    Evidence Co-IP, ubiquitination assays, Notch4-deficient mice with M. tuberculosis infection and bacterial burden measurement

    PMID:29228365

    Open questions at the time
    • Whether this cytoplasmic function is independent of nuclear Notch4 signaling was not fully dissected
    • Relevance to human tuberculosis immunity was not established
  18. 2020 High

    Two key advances: direct ChIP confirmation that NOTCH4-ICD binds SLUG and GAS1 promoters to maintain mesenchymal cancer stem cell quiescence in TNBC; and discovery that Notch4 on Tregs drives inflammatory reprogramming via Wnt/Hippo-dependent GDF15 production activating ILC2s.

    Evidence ChIP and dual-luciferase assays for SLUG/GAS1 promoters in TNBC; Treg-specific Notch4 conditional knockout in allergen challenge models with pathway inhibitors

    PMID:32104513 PMID:32929274

    Open questions at the time
    • How Notch4 engages Wnt/Hippo pathways in Tregs was not mechanistically defined at the protein level
    • Whether GAS1-mediated quiescence and SLUG-driven EMT are coordinated or independent was unclear
  19. 2021 High

    Notch4 was demonstrated to regulate lymphangiogenesis through a non-canonical (RBP-Jκ-independent) pathway distinct from Notch1, and separately shown to suppress Treg-mediated tissue repair by inhibiting amphiregulin, with therapeutic anti-Notch4 antibody rescuing viral infection morbidity.

    Evidence Notch4-null mice with lymphatic vessel analysis and DNMAML canonical pathway control; Treg-specific Notch4 cKO and anti-Notch4 antibody in influenza model

    PMID:33915108 PMID:34665379

    Open questions at the time
    • Identity of the non-canonical Notch4 effector in lymphangiogenesis was unknown
    • Whether anti-Notch4 therapy has off-target vascular effects was not assessed
  20. 2022 High

    JAG1-NOTCH4 was identified as a mechanosensitive pathway activated by disturbed blood flow to promote atherosclerosis, and separately Notch4 was found to physically interact with ERK/JNK/P38 MAPK to promote pulmonary artery smooth muscle cell proliferation under hypoxia.

    Evidence Disturbed flow models with EC-specific Jag1 cKO and scRNA-seq; co-IP of Notch4 with MAPK in PASMCs plus AAV1-mediated in vivo silencing in hypoxic pulmonary hypertension rats

    PMID:35016680 PMID:36044575

    Open questions at the time
    • Whether NOTCH4 is the sole Notch receptor mediating flow-dependent atherogenesis was not resolved
    • Structural basis for Notch4-MAPK interaction was not determined
    • Whether the smooth muscle cell function is cleavage-dependent was not tested

Open questions

Synthesis pass · forward-looking unresolved questions
  • Major unresolved questions include: the structural basis for Notch4's unique properties (cis-inhibition, lack of canonical ligand response); the identity of non-canonical effectors in lymphangiogenesis and mammary tumorigenesis; whether and how full-length versus cleaved Notch4 functions are coordinated in vivo; and the therapeutic window for Notch4-targeted interventions given its opposing roles in vascular, immune, and cancer contexts.
  • No crystal structure of Notch4 extracellular or intracellular domains exists
  • Comprehensive interactome of Notch4-ICD in primary endothelial cells has not been determined
  • Context-dependent switching between pro- and anti-EMT effects lacks a unified mechanistic explanation

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140110 transcription regulator activity 6 GO:0098772 molecular function regulator activity 4 GO:0060090 molecular adaptor activity 2
Localization
GO:0005634 nucleus 4 GO:0005886 plasma membrane 3
Pathway
R-HSA-162582 Signal Transduction 9 R-HSA-1643685 Disease 7 R-HSA-1266738 Developmental Biology 6 R-HSA-74160 Gene expression (Transcription) 5 R-HSA-168256 Immune System 4
Partners
DLL4JAG1MAPK1RBPJSMAD3TAK1TRAF6YWHAZ
Complex memberships
RBP-Jκ/CBF1 transcription complex

Evidence

Reading pass · 38 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1996 Notch4 (int-3) is specifically expressed in endothelial cells during embryonic and adult development, and loss of the extracellular domain (as in the int-3 oncogene) leads to constitutive activation of the intracellular signaling domain, causing neoplastic transformation of mammary epithelial cells. In situ hybridization, cDNA cloning, transgenic mouse models Development High 8681805
1992 Expression of the truncated int-3/Notch4 intracellular domain (encoding the intracellular portion homologous to Drosophila Notch) in transgenic mice causes mammary gland differentiation arrest and adenocarcinoma formation, establishing that the intracellular domain is a constitutively active oncogenic form. Transgenic mouse model, histology, MMTV promoter-driven expression Genes & development High 1372276
1992 The int-3/Notch4 intracellular domain protein contains six cdc10/ankyrin repeats homologous to Drosophila Notch, and expression of this domain in HC11 mouse mammary epithelial cells induces anchorage-independent growth in soft agar, demonstrating oncogenic transformation. In vitro soft-agar transformation assay, sequence analysis Journal of virology High 1312643
2001 Endothelial-specific expression of activated Notch4 under the Flk1 promoter in mice causes embryonic lethality with vascular patterning defects including restricted vasculature, dilated vessels, disorganized networks, and compromised vessel-wall integrity, implicating Notch4 in vessel remodeling. Knock-in transgenic mouse model, histology, whole-mount analysis PNAS High 11344305
2002 Constitutively active Notch4 inhibits VEGF-induced angiogenesis and endothelial sprouting in vitro and in vivo by promoting beta1-integrin activation (high-affinity conformation) and increased adhesion to collagen, without increasing surface expression of beta1-integrins. In vitro endothelial sprouting assay, chick CAM assay, flow cytometry for integrin activation, function-activating antibodies Molecular and cellular biology High 11909975
2003 Constitutively active Notch4 intracellular domain inhibits endothelial apoptosis via two distinct pathways: (1) RBP-Jkappa-dependent inhibition of JNK-mediated apoptosis, and (2) RBP-Jkappa-independent upregulation of Bcl-2. Overexpression of Notch4-ICD and deletion mutants, RBP-Jkappa reporter assays, apoptosis assays, Western blot Journal of Biological Chemistry High 14701863
2004 Notch4-induced inhibition of endothelial sprouting requires the ankyrin repeats but not the RAM domain or C-terminal region, and involves both RBP-Jkappa-dependent and -independent signaling; the ankyrin repeats alone are sufficient to partially inhibit sprouting and upregulate RBP-Jkappa target genes. Deletion mutant analysis, quantitative endothelial sprouting assay, RBP-Jkappa reporter assays Blood High 15187023
2005 Notch4 intracellular domain (ICD) binds Smad3 (with higher affinity than Smad2 or Smad4) via the Smad3 MH2 domain, independent of the RAM23 region, and inhibits TGF-beta/Activin-Smad signaling, rendering cells resistant to TGF-beta-induced growth inhibition. Co-immunoprecipitation, deletion analysis, TGF-beta reporter luciferase assays, RT-PCR, Western blot, gamma-secretase inhibitor treatment Oncogene High 16007227
2005 NOTCH4 transcription in vascular endothelial cells is controlled by AP-1 complexes occupying the NOTCH4 chromatin; intron 1 or upstream sequences are required for endothelial-specific expression in vivo, and vascular angiogenic factors activate AP-1 to reprogram NOTCH4 from a repressed to transcriptionally active state. Chromatin immunoprecipitation (ChIP), transgenic mouse reporter assays, RNA-FISH, transfection assays, histone modification analysis Molecular and Cellular Biology High 15684396
2005 In adult mouse endothelium, constitutively active Notch4 (int3) causes arteriovenous malformations with blood vessel enlargement, AV shunting, arterialization (ectopic venous ephrinB2 expression, increased smooth muscle cells, upregulated Notch signaling), and these defects are fully reversible upon repression of int3 expression. Tetracycline-repressible endothelial transgenic system, histology, immunofluorescence PNAS High 15994223
2006 VEGF upregulates DLL4 and presenilin, leading to activation of Notch4, which in turn upregulates ephrinB2 and downregulates EphB4 in venous endothelial cells; pharmacological inhibition of presenilin or soluble DLL4 blocks VEGF-induced Notch4 activation and venous endothelial differentiation and migration. In vitro endothelial cell assays, Western blot, immunohistochemistry in transgenic tumor model, pharmacological inhibition Cancer research Medium 16951162
2007 Glucocorticoid receptor (GR) and AP-1 synergistically activate NOTCH4 transcription in endothelial cells via a composite response element (imperfect GRE + AP-1 motif) in the NOTCH4 promoter; AP-1 stabilizes GR occupancy, and together they reprogram NOTCH4 chromatin from repressed to active in multipotent cells. ChIP assay, promoter reporter assays, composite element mutagenesis, histone modification analysis Molecular and Cellular Biology High 17220278
2008 Constitutively active Notch4 expressed in brain endothelium from birth causes hallmarks of brain arteriovenous malformations including cerebral AV shunting, vessel enlargement, hemorrhage, and neuronal cell death; repression of Notch4 resolves ataxia and reverses disease, indicating Notch4 is required to sustain as well as initiate the AVM phenotype. Tetracycline-repressible endothelial-specific transgenic system, MRI, histology, behavioral assays PNAS High 18667694
2008 Notch4 knockdown in endothelial cells increases VCAM-1 expression and promotes apoptosis, while Notch4 and Hes1 are required to maintain EC quiescence and for endothelial injury repair; impaired Notch4/Hes1 activity is associated with transplant arteriosclerosis. siRNA knockdown, CBF1 reporter assays, adhesion molecule expression, apoptosis assays, allograft model Arteriosclerosis, Thrombosis, and Vascular Biology Medium 18802018
2008 Notch4 signaling in mammary glands is mediated primarily through interaction of the intracellular domain with RBP-Jkappa (CBF1): conditional Rbpj knockout rescues the mammary gland developmental arrest induced by Wap-Int3, but mammary tumor development occurs independently of RBP-Jkappa interaction. Conditional Rbpj knockout in mammary glands, Wap-Cre system, mammary transplantation assays Oncogene High 18836481
2010 Notch4 signaling activity is 8-fold higher in breast cancer stem cell-enriched populations compared with differentiated cells (versus 4-fold lower for Notch1); pharmacological or genetic inhibition of Notch4 completely inhibits tumor initiation in vivo, implicating Notch4 as the primary Notch receptor maintaining breast cancer stem cell activity. Stem cell enrichment (anoikis-resistance, ESA+/CD44+/CD24low), Notch activity reporter assays, GSI and siRNA inhibition, in vivo tumor formation assay Cancer research High 20068161
2010 Notch4 is specifically required for Nodal expression in aggressive melanoma cells via an RBPJ-dependent Nodal enhancer element, placing Notch4 upstream of Nodal in the signaling cascade; Notch4-mediated vasculogenic mimicry and anchorage-independent growth are partly dependent on Nodal. Notch4 knockdown, RBPJ-dependent reporter assays, expression correlation in multiple cell lines, rescue experiments Cancer research Medium 21159651
2012 Normalization of Notch4 signaling in mouse brain converts large-caliber high-flow AV shunts to capillary-like vessels and reverses tissue hypoxia; this structural regression requires restoration of EphB4 receptor expression by venous ECs and occurs via vessel narrowing without loss of ECs. 4D two-photon imaging through cranial windows, Notch4* repression, EphB4 expression analysis, blood flow measurement Science Translational Medicine High 22261032
2013 PKCα overexpression selectively increases Notch4 (but not Notch1) expression via AP-1 occupancy at the Notch4 promoter; Notch4 intracellular domain promotes estrogen-independent, tamoxifen-resistant breast cancer growth and activates pathways associated with endocrine resistance. ChIP for AP-1 on Notch4 promoter, siRNA knockdown, Notch4-IC expression, gene expression profiling, in vivo xenograft with gamma-secretase inhibitor Oncogenesis High 23917222
2013 YB-1 (Y-box binding protein-1) transcriptionally regulates Notch4: ChIP-on-chip shows 12-fold enrichment of YB-1 binding at the Notch4 promoter, YB-1 siRNA decreases Notch4 mRNA, and constitutively active YB-1 increases Notch4 mRNA. Upstream, RSK phosphorylates YB-1 (S102), and RSK inhibition with luteolin suppresses Notch4 levels and reduces tumor-initiating cells. ChIP-on-Chip, siRNA knockdown, overexpression constructs, in vitro kinase assay, Notch4 mRNA measurement Oncotarget High 23593654
2014 NOTCH4 did not signal in response to canonical ligand activation in cultured cells, unlike other Notch receptors; furthermore, NOTCH4 inhibits signaling from NOTCH1 in cis (the first report of cis-inhibition by another Notch receptor), and this inhibition is mediated by the NOTCH4 extracellular domain. In vivo, Notch4 null mice show slightly delayed vessel growth in the retina. Notch signaling reporter assays in cultured cells, NOTCH4 knockout mouse generation (full coding region deletion), cis-inhibition constructs, retinal vascular imaging Biochimica et Biophysica Acta High 24667410
2014 Brain AVMs arise from enlargement of pre-existing capillary-diameter microvessels accompanied by increased individual endothelial cell area (not increased EC number/proliferation); AV shunting begins promptly after Notch4* expression, and alterations in Notch signaling in ECs of all vessels (not arteries alone) drive AVM formation, indicating Notch4 functions in microvasculature/veins. Time-lapse two-photon imaging through cranial windows in Notch4* mice, EC area/number quantification, cell-type-specific Notch manipulation PNAS High 25468970
2015 AKT phosphorylates Notch4 intracellular domain (ICD) at four distinct sites in vitro and in vivo; this phosphorylation is regulated by growth factors and PI3K, creates binding sites for 14-3-3 proteins, and restricts phosphorylated Notch4-ICD to the cytoplasm, thereby negatively regulating Notch4 nuclear signaling. In vitro kinase assay, site-directed mutagenesis, co-immunoprecipitation with 14-3-3, PI3K inhibitor treatment, subcellular fractionation Scientific Reports High 25740432
2015 JAG1-NOTCH4 activation by anti-estrogen treatment (tamoxifen/fulvestrant) increases breast cancer stem cell (BCSC) activity; targeting NOTCH4 reverses this increase and reduces BCSC activity in patient-derived xenograft tumors with acquired tamoxifen resistance. Patient-derived samples, PDX models, NOTCH4 targeting, ALDH1 activity, mammosphere assay Cell Reports High 26387946
2016 Constitutively active Notch4 ICD (N4ICD) in melanoma cells triggers a mesenchymal-to-epithelial-like switch and suppresses invasion/migration; mechanistically, N4ICD induces Hey1 and Hey2 transcription factors which directly bind the promoters of Snail2 and Twist1 and repress their transcription, as shown by EMSA and luciferase assays. N4ICD overexpression, EMSA, luciferase reporter assays, invasion/migration assays, in vivo tumor growth assay Cancer Research High 26801977
2017 NOTCH4 transcriptionally activates HEY1 in HNSCC, and NOTCH4-HEY1 pathway activation promotes EMT (decreased E-cadherin, increased vimentin/fibronectin/Twist1/SOX2), proliferation, cisplatin resistance, and invasion. TCGA analysis, in vitro proliferation/apoptosis/cell-cycle assays, siRNA knockdown, expression analysis Clinical Cancer Research Medium 29146722
2017 NOTCH4 expression is negatively controlled by RUNX1 via a novel regulatory DNA element in the NOTCH4 locus; NOTCH4 inactivation by CRISPR-Cas9 in human iPSCs enhances megakaryopoiesis, establishing NOTCH4 as a RUNX1 target gene that negatively regulates megakaryocyte differentiation. Integrative genomic analysis of FPD-iPSCs, ChIP for RUNX1 binding, CRISPR-Cas9 NOTCH4 knockout, MK differentiation assays from iPSCs and CD34+ cells Blood High 29101237
2018 Notch4 intracellular domain interacts with TAK1 and inhibits its activation; furthermore, Notch4-ICD prevents TRAF6 autoubiquitination and suppresses TRAF6-mediated TAK1 polyubiquitination, thereby negatively regulating M. tuberculosis-induced proinflammatory cytokine production. Co-immunoprecipitation, ubiquitination assays, Notch4-deficient mice, bacterial burden measurement Journal of Infectious Diseases High 29228365
2020 NOTCH4 transcriptionally upregulates SLUG and GAS1 by directly binding their promoters (confirmed by dual-luciferase reporter and ChIP assays), promoting EMT and quiescence in triple-negative breast cancer mesenchymal-like cancer stem cells. RNA-seq, dual-luciferase reporter assays, chromatin immunoprecipitation, stable overexpression and knockdown, mammosphere and chemoresistance assays Theranostics High 32104513
2020 Notch4 on lung tissue regulatory T cells (induced by IL-6/STAT3) subverts Treg cells into TH2 and TH17 effector T cells via Wnt and Hippo pathway-dependent mechanisms; Wnt activation induces GDF15 expression in Treg cells, which activates group 2 innate lymphoid cells in a feed-forward inflammatory loop. Treg-specific Notch4 conditional knockout mice, in vivo allergen challenge models, Wnt/Hippo pathway inhibitors, GDF15 measurement, cytokine assays Nature immunology High 32929274
2021 Notch4 on circulating regulatory T cells suppresses IL-18-induced amphiregulin production (a tissue repair cytokine); deletion of Notch4 in Treg cells or anti-Notch4 antibody therapy normalizes dysregulated innate immunity and rescues disease morbidity in viral RNA analog and influenza H1N1 infection models. Treg-specific Notch4 conditional knockout, anti-Notch4 antibody therapy, amphiregulin measurement, humanized mice, IL-18 stimulation assays Immunity High 33915108
2021 DLL4 activation of Notch induces distinct subsets of Notch effectors and lymphatic genes in lymphatic endothelial cells (LECs), with Notch1 and Notch4 differentially regulating these genes; Notch4-null mice show increased closure of lymphangiogenic fronts, reduced vessel caliber, and reduced branching; Notch4 activation suppresses LEC migration more than Notch1, and loss of Notch4 does not affect canonical Notch signaling, establishing Notch4 as acting through a non-canonical pathway in lymphangiogenesis. Notch4 null mouse generation, embryonic lymphatic analysis, LEC migration wound assay, DNMAML dominant-negative canonical Notch inhibition, DLL4 activation assays Angiogenesis High 34665379
2021 NOTCH4 inhibits IFN-γ signaling in macrophages by favoring STAT3 over STAT1 phosphorylation (without affecting their expression), reducing STAT1-dependent transcription; NOTCH4 also reduces NF-κB transcriptional activity downstream of LPS/TLR4 signaling; HES1 mediates at least part of the STAT3 enhancement by NOTCH4. NOTCH4 overexpression in macrophages, STAT1/STAT3 phosphorylation assays, NF-κB reporter assays, cytokine measurement, gene expression analysis Frontiers in Immunology Medium 34925319
2022 JAG1-NOTCH4 signaling is activated by disturbed blood flow in endothelial cells; EC-specific Jag1 deletion in mice reduces atherosclerosis at sites of disturbed flow; single-cell RNA sequencing shows Jag1 suppresses EC subsets that proliferate and migrate, establishing JAG1-NOTCH4 as a mechanosensor pathway promoting atherosclerosis. Disturbed flow models (porcine/murine arteries and cultured human coronary artery EC), light-sheet imaging, EC-specific Jag1 conditional knockout, single-cell RNA sequencing Science Advances High 36044575
2022 Notch4 promotes pulmonary artery smooth muscle cell proliferation and migration and inhibits apoptosis under hypoxia via interaction with ERK, JNK, and P38 MAPK; co-immunoprecipitation confirms physical interaction between Notch4 and ERK/JNK/P38; AAV1-mediated Notch4 silencing in rats reduces hypoxia-induced right ventricular systolic pressure and pulmonary vascular remodeling. Co-immunoprecipitation, siRNA knockdown, overexpression, cell viability/proliferation/apoptosis/migration assays, AAV1-mediated in vivo silencing in HPH rats Respiratory Research High 35016680
2011 PEA3 (an ETS transcription factor) activates NOTCH4 transcription in breast cancer cells; PEA3 recruitment to the NOTCH4 promoter is c-JUN/AP-1-dependent (unlike PEA3 recruitment to Notch-1 which is AP-1-independent), confirmed by ChIP and Notch4 luciferase reporter assays. ChIP for PEA3 on NOTCH4 promoter, siRNA knockdown, Notch4 luciferase reporter, c-Jun siRNA/TAM-67 epistasis Breast Cancer Research High 21679465
1998 The minimal domain of Notch4(int-3) required to inhibit mammary epithelial branching morphogenesis consists of the CBF-1 interaction domain and the cdc10 repeat domain; Wnt-1 can overcome Notch4-mediated inhibition of branching, demonstrating opposing roles of Notch and Wnt pathways in mammary morphogenesis. Notch4 deletion constructs transfected into TAC-2 mammary epithelial cells, 3D collagen gel branching morphogenesis assay, co-expression of Wnt-1 Developmental Biology High 9576833
2000 Both constitutively active Notch4/int-3 and Jagged-1 induce microvessel-like structures in rat brain endothelial cells (RBE4) with morphological and biochemical brain endothelial properties; this correlates with induction of endogenous Notch4 and Jagged-1, while full-length Notch4 does not affect RBE4 cells. Transfection of RBE4 cells, morphological analysis, gene expression analysis of endogenous Notch4 and Jagged-1 Microvascular Research Medium 10964583

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1996 Notch4/int-3, a mammary proto-oncogene, is an endothelial cell-specific mammalian Notch gene. Development (Cambridge, England) 427 8681805
2010 Regulation of breast cancer stem cell activity by signaling through the Notch4 receptor. Cancer research 420 20068161
1992 Expression of an activated Notch-related int-3 transgene interferes with cell differentiation and induces neoplastic transformation in mammary and salivary glands. Genes & development 320 1372276
1992 Mouse mammary tumor gene int-3: a member of the notch gene family transforms mammary epithelial cells. Journal of virology 237 1312643
2001 Vascular patterning defects associated with expression of activated Notch4 in embryonic endothelium. Proceedings of the National Academy of Sciences of the United States of America 227 11344305
1994 Three genes in the human MHC class III region near the junction with the class II: gene for receptor of advanced glycosylation end products, PBX2 homeobox gene and a notch homolog, human counterpart of mouse mammary tumor gene int-3. Genomics 210 7835890
1997 The mouse mammary tumor associated gene INT3 is a unique member of the NOTCH gene family (NOTCH4). Oncogene 166 9150355
2005 Endothelial expression of constitutively active Notch4 elicits reversible arteriovenous malformations in adult mice. Proceedings of the National Academy of Sciences of the United States of America 164 15994223
2015 Anti-estrogen Resistance in Human Breast Tumors Is Driven by JAG1-NOTCH4-Dependent Cancer Stem Cell Activity. Cell reports 160 26387946
1996 Expression of a truncated Int3 gene in developing secretory mammary epithelium specifically retards lobular differentiation resulting in tumorigenesis. Cancer research 155 8620493
2002 Activated Notch4 inhibits angiogenesis: role of beta 1-integrin activation. Molecular and cellular biology 140 11909975
2006 The role of the vascular endothelial growth factor-Delta-like 4 ligand/Notch4-ephrin B2 cascade in tumor vessel remodeling and endothelial cell functions. Cancer research 138 16951162
1987 A new common integration region (int-3) for mouse mammary tumor virus on mouse chromosome 17. Journal of virology 118 3023699
2008 Endothelial Notch4 signaling induces hallmarks of brain arteriovenous malformations in mice. Proceedings of the National Academy of Sciences of the United States of America 113 18667694
1998 Notch4 and Wnt-1 proteins function to regulate branching morphogenesis of mammary epithelial cells in an opposing fashion. Developmental biology 111 9576833
2021 Notch4 signaling limits regulatory T-cell-mediated tissue repair and promotes severe lung inflammation in viral infections. Immunity 109 33915108
2020 A regulatory T cell Notch4-GDF15 axis licenses tissue inflammation in asthma. Nature immunology 104 32929274
2010 Regulation of the embryonic morphogen Nodal by Notch4 facilitates manifestation of the aggressive melanoma phenotype. Cancer research 101 21159651
2003 Notch4 inhibits endothelial apoptosis via RBP-Jkappa-dependent and -independent pathways. The Journal of biological chemistry 98 14701863
2011 Notch-3 and Notch-4 signaling rescue from apoptosis human B-ALL cells in contact with human bone marrow-derived mesenchymal stromal cells. Blood 97 21602525
2007 Aberrant Notch3 and Notch4 expression in human hepatocellular carcinoma. Liver international : official journal of the International Association for the Study of the Liver 95 17696940
1995 Constitutive expression of a truncated INT3 gene in mouse mammary epithelium impairs differentiation and functional development. Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research 90 7544153
2012 Genome-wide association study of age-related macular degeneration identifies associated variants in the TNXB-FKBPL-NOTCH4 region of chromosome 6p21.3. Human molecular genetics 81 22694956
2005 Notch4 intracellular domain binding to Smad3 and inhibition of the TGF-beta signaling. Oncogene 81 16007227
2000 Expression of an activated Notch4(int-3) oncoprotein disrupts morphogenesis and induces an invasive phenotype in mammary epithelial cells in vitro. International journal of cancer 80 10797286
2010 Inflammation dysregulates Notch signaling in endothelial cells: implication of Notch2 and Notch4 to endothelial dysfunction. Biochemical pharmacology 78 20643108
2017 The NOTCH4-HEY1 Pathway Induces Epithelial-Mesenchymal Transition in Head and Neck Squamous Cell Carcinoma. Clinical cancer research : an official journal of the American Association for Cancer Research 75 29146722
2014 Constitutively active Notch4 receptor elicits brain arteriovenous malformations through enlargement of capillary-like vessels. Proceedings of the National Academy of Sciences of the United States of America 74 25468970
2011 Notch-1 and Notch-4 biomarker expression in triple-negative breast cancer. International journal of surgical pathology 72 22084425
2012 Notch4 normalization reduces blood vessel size in arteriovenous malformations. Science translational medicine 70 22261032
2020 NOTCH4 maintains quiescent mesenchymal-like breast cancer stem cells via transcriptionally activating SLUG and GAS1 in triple-negative breast cancer. Theranostics 69 32104513
2017 Essential role of Notch4/STAT3 signaling in epithelial-mesenchymal transition of tamoxifen-resistant human breast cancer. Cancer letters 65 28108315
2005 Molecular determinants of NOTCH4 transcription in vascular endothelium. Molecular and cellular biology 63 15684396
2008 Impaired Notch4 activity elicits endothelial cell activation and apoptosis: implication for transplant arteriosclerosis. Arteriosclerosis, thrombosis, and vascular biology 60 18802018
2004 Constitutively active Notch4 promotes early human hematopoietic progenitor cell maintenance while inhibiting differentiation and causes lymphoid abnormalities in vivo. Blood 60 15231576
1997 Proto-oncogene of int-3, a mouse Notch homologue, is expressed in endothelial cells during early embryogenesis. Genes to cells : devoted to molecular & cellular mechanisms 60 9189758
2014 Nicastrin and Notch4 drive endocrine therapy resistance and epithelial to mesenchymal transition in MCF7 breast cancer cells. Breast cancer research : BCR 59 24919951
2005 Unique patterns of Notch1, Notch4 and Jagged1 expression in ovarian vessels during folliculogenesis and corpus luteum formation. Gene expression patterns : GEP 59 15939383
2013 Crosstalk between PKCα and Notch-4 in endocrine-resistant breast cancer cells. Oncogenesis 57 23917222
2000 Notch4 and Jagged-1 induce microvessel differentiation of rat brain endothelial cells. Microvascular research 57 10964583
2011 NOTCH-1 and NOTCH-4 are novel gene targets of PEA3 in breast cancer: novel therapeutic implications. Breast cancer research : BCR 54 21679465
2012 Withaferin A causes activation of Notch2 and Notch4 in human breast cancer cells. Breast cancer research and treatment 52 22965833
2021 Identification of NOTCH4 mutation as a response biomarker for immune checkpoint inhibitor therapy. BMC medicine 50 34284787
2019 FKBPL and its peptide derivatives inhibit endocrine therapy resistant cancer stem cells and breast cancer metastasis by downregulating DLL4 and Notch4. BMC cancer 50 30975104
2014 NOTCH4 is a potential therapeutic target for triple-negative breast cancer. Anticancer research 50 24403446
2017 Notch-4 silencing inhibits prostate cancer growth and EMT via the NF-κB pathway. Apoptosis : an international journal on programmed cell death 49 28374086
2014 Notch4 reveals a novel mechanism regulating Notch signal transduction. Biochimica et biophysica acta 48 24667410
2013 Luteolin is a novel p90 ribosomal S6 kinase (RSK) inhibitor that suppresses Notch4 signaling by blocking the activation of Y-box binding protein-1 (YB-1). Oncotarget 48 23593654
2016 Notch4+ cancer stem-like cells promote the metastatic and invasive ability of melanoma. Cancer science 45 27234159
2016 Notch4 Signaling Induces a Mesenchymal-Epithelial-like Transition in Melanoma Cells to Suppress Malignant Behaviors. Cancer research 44 26801977
2008 Rbpj conditional knockout reveals distinct functions of Notch4/Int3 in mammary gland development and tumorigenesis. Oncogene 44 18836481
2001 Failure to confirm NOTCH4 association with schizophrenia in a large population-based sample from Scotland. Nature genetics 44 11381258
2004 Mammary development and tumorigenesis in mice expressing a truncated human Notch4/Int3 intracellular domain (h-Int3sh). Oncogene 41 15531924
2002 A family-based and case-control association study of the NOTCH4 gene and schizophrenia. Molecular psychiatry 40 11803454
2001 NOTCH4 gene polymorphism and susceptibility to schizophrenia and schizoaffective disorder. Neuroscience letters 40 11239712
2004 Notch4-induced inhibition of endothelial sprouting requires the ankyrin repeats and involves signaling through RBP-Jkappa. Blood 38 15187023
1998 Cloning, characterization, and the complete 56.8-kilobase DNA sequence of the human NOTCH4 gene. Genomics 37 9693032
2018 The association between Notch4 expression, and clinicopathological characteristics and clinical outcomes in patients with breast cancer. Oncology letters 36 29805613
2017 Human NOTCH4 is a key target of RUNX1 in megakaryocytic differentiation. Blood 36 29101237
2015 Hepatitis B virus X protein activates Notch signaling by its effects on Notch1 and Notch4 in human hepatocellular carcinoma. International journal of oncology 34 26530164
2000 Imbalanced expression of TAN-1 and human Notch4 in endometrial cancers. International journal of oncology 34 11078798
2022 Notch4 mediates vascular remodeling via ERK/JNK/P38 MAPK signaling pathways in hypoxic pulmonary hypertension. Respiratory research 33 35016680
2010 Chronic exposure to nicotine and saquinavir decreases endothelial Notch-4 expression and disrupts blood-brain barrier integrity. Journal of neurochemistry 33 20722969
2021 NOTCH4 Exhibits Anti-Inflammatory Activity in Activated Macrophages by Interfering With Interferon-γ and TLR4 Signaling. Frontiers in immunology 32 34925319
2004 Interaction between NOTCH4 and catechol-O-methyltransferase genotypes in schizophrenia patients with poor response to typical neuroleptics. Pharmacogenetics 32 15115916
2014 NOTCH1, NOTCH3, NOTCH4, and JAG2 protein levels in human endometrial cancer. Medicina (Kaunas, Lithuania) 31 25060200
2021 Rare variant analysis in eczema identifies exonic variants in DUSP1, NOTCH4 and SLC9A4. Nature communications 30 34785669
2019 Quartz crystal microbalance biosensor for label-free MDA MB 231 cancer cell detection via notch-4 receptor. Talanta 30 31357373
2014 Notch4 promotes gastric cancer growth through activation of Wnt1/β-catenin signaling. Molecular and cellular biochemistry 30 25511451
2012 A re-review of the association between the NOTCH4 locus and schizophrenia. American journal of medical genetics. Part B, Neuropsychiatric genetics : the official publication of the International Society of Psychiatric Genetics 30 22488909
2010 Notch-4 contributes to the metastasis of salivary adenoid cystic carcinoma. Oncology reports 30 20596622
2001 The (CTG)n polymorphism in the NOTCH4 gene is not associated with schizophrenia in Japanese individuals. BMC psychiatry 30 11407996
2022 The NOTCH4-GATA4-IRG1 axis as a novel target in early-onset colorectal cancer. Cytokine & growth factor reviews 29 35941043
2020 Androgen receptor suppresses vasculogenic mimicry in hepatocellular carcinoma via circRNA7/miRNA7-5p/VE-cadherin/Notch4 signalling. Journal of cellular and molecular medicine 29 33118329
2018 Notch4 Negatively Regulates the Inflammatory Response to Mycobacterium tuberculosis Infection by Inhibiting TAK1 Activation. The Journal of infectious diseases 29 29228365
2003 Notch4, a non-HLA gene in the MHC is strongly associated with the most severe form of alopecia areata. Human genetics 29 12589427
2022 JAG1-NOTCH4 mechanosensing drives atherosclerosis. Science advances 28 36044575
2016 NOTCH4 gene polymorphisms as potential risk factors for brain arteriovenous malformation development and hemorrhagic presentation. Journal of neurosurgery 28 27231971
2003 NOTCH4 and the frontal lobe in schizophrenia. American journal of medical genetics. Part B, Neuropsychiatric genetics : the official publication of the International Society of Psychiatric Genetics 28 12627456
2003 NOTCH4 gene promoter polymorphism is associated with the age of onset in schizophrenia. Psychiatric genetics 27 12782960
2015 AKT and 14-3-3 regulate Notch4 nuclear localization. Scientific reports 26 25740432
2012 Active form Notch4 promotes the proliferation and differentiation of 3T3-L1 preadipocytes. Biochemical and biophysical research communications 25 23237809
2009 Constitutively active endothelial Notch4 causes lung arteriovenous shunts in mice. American journal of physiology. Lung cellular and molecular physiology 25 19933399
2021 Targeting Notch4 in Cancer: Molecular Mechanisms and Therapeutic Perspectives. Cancer management and research 24 34526819
2004 NOTCH4 gene haplotype is associated with schizophrenia in African Americans. Biological psychiatry 24 14732589
2004 Expression of constitutively active Notch4 (Int-3) modulates myeloid proliferation and differentiation and promotes expansion of hematopoietic progenitors. Leukemia 24 14961038
2015 The role of polycomb group protein Bmi-1 and Notch4 in breast cancer stem cell inhibition by benzyl isothiocyanate. Breast cancer research and treatment 23 25663545
1997 Gene organization of human NOTCH4 and (CTG)n polymorphism in this human counterpart gene of mouse proto-oncogene Int3. Gene 23 9168133
1994 Insertional mutation of int protooncogenes in the mammary tumors of a new strain of mice derived from the wild in China: normal- and tumor-tissue-specific expression of int-3 transcripts. Virology 23 8030284
2022 A common IL-4 receptor variant promotes asthma severity via a Treg cell GRB2-IL-6-Notch4 circuit. Allergy 22 35841382
2004 A NOTCH4 association with multiple sclerosis is secondary to HLA-DR*1501. Tissue antigens 22 14651518
2022 Expression of NOTCH1, NOTCH4, HLA-DMA and HLA-DRA is synergistically associated with T cell exclusion, immune checkpoint blockade efficacy and recurrence risk in ER-negative breast cancer. Cellular oncology (Dordrecht, Netherlands) 21 35543859
2021 Unique functions for Notch4 in murine embryonic lymphangiogenesis. Angiogenesis 21 34665379
2019 Vascular endothelial growth factor 165 inhibits pro-fibrotic differentiation of stromal cells via the DLL4/Notch4/smad7 pathway. Cell death & disease 21 31515487
2015 The vascular delta-like ligand-4 (DLL4)-Notch4 signaling correlates with angiogenesis in primary glioblastoma: an immunohistochemical study. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 21 26472724
2008 Alterations in the Notch4 pathway in cerebral endothelial cells by the HIV aspartyl protease inhibitor, nelfinavir. BMC neuroscience 20 18302767
2007 Glucocorticoid and growth factor synergism requirement for Notch4 chromatin domain activation. Molecular and cellular biology 20 17220278
2006 A review and re-evaluation of an association between the NOTCH4 locus and schizophrenia. American journal of medical genetics. Part B, Neuropsychiatric genetics : the official publication of the International Society of Psychiatric Genetics 20 16894623
2005 Five NOTCH4 polymorphisms show weak evidence for association with schizophrenia: evidence from meta-analyses. Schizophrenia research 20 15653273
2003 Family-based association study of the NOTCH4 gene in schizophrenia using Japanese and Chinese samples. Biological psychiatry 20 12873802