{"gene":"VEGFA","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":1993,"finding":"VEGF-A (VPF/VEGF) acts directly on endothelial cells via two high-affinity tyrosine kinase receptors to activate phospholipase C and induce intracellular calcium transients; it is secreted as a 34–42 kDa heparin-binding, dimeric, disulfide-bonded glycoprotein.","method":"Receptor binding assays, functional in vitro assays (phospholipase C activation, calcium transients), biochemical characterization","journal":"Cancer metastasis reviews","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — direct in vitro biochemical assays with multiple orthogonal methods, replicated across multiple labs over time","pmids":["8281615"],"is_preprint":false},{"year":2002,"finding":"Akt signaling is both necessary and sufficient for VEGF-A-induced vascular permeability in vivo; dominant-negative Akt blocks VEGF-induced permeability, and constitutively active Akt mimics it; this Akt-mediated permeability requires eNOS activity.","method":"Adenovirus-mediated gene transfer of dominant-negative and constitutively active Akt in vivo (Miles assay), eNOS inhibitor (L-NAME) blockade","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vivo gain- and loss-of-function with mechanistic inhibitor validation, single lab but multiple orthogonal approaches","pmids":["12459464"],"is_preprint":false},{"year":2005,"finding":"VEGF-A enhances endothelial PDGF-B expression; combined with FGF-2 (which enhances mural PDGFRβ expression), this directs endogenous PDGF-B–PDGFRβ signaling to recruit mural cells and form functional neovasculature; abrogation by anti-PDGFRβ antibody confirmed this mechanism.","method":"In vitro VEGFR2+ ESC-derived cell stimulation assays, Matrigel plug assay in vivo, neutralizing antibody blockade","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal in vitro and in vivo experiments with neutralizing antibody epistasis, single lab","pmids":["16105884"],"is_preprint":false},{"year":2005,"finding":"VEGF-A stimulates chemotactic migration of human mesenchymal progenitor cells via VEGFR-1 (Flt-1); VEGFR-1 and VEGFR-2 are activated upon ligand stimulation in these cells, and the effect is mediated by VEGFR-1 since PlGF-1 (a VEGFR-1-selective ligand) but not VEGF-E or VEGF-C produced the same effect.","method":"In vitro kinase assay, Boyden chamber chemotaxis assay, quantitative RT-PCR for receptor expression","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — receptor activation confirmed by in vitro kinase assay, isoform selectivity used as epistatic evidence, single lab","pmids":["16005848"],"is_preprint":false},{"year":2005,"finding":"VEGF-A overexpression in primary tumors induces lymphangiogenesis in sentinel lymph nodes even before metastasis occurs; VEGF-A acts on VEGFR-2-expressing lymphatic vessels to drive both tumor-associated and sentinel lymph node lymphangiogenesis.","method":"VEGF-A skin-specific transgenic mice, chemically induced carcinogenesis, immunohistochemistry for lymphatic markers","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — transgenic gain-of-function in vivo model with histological readout, well-controlled study","pmids":["15809353"],"is_preprint":false},{"year":2005,"finding":"VEGF-A gene targeting in thymus epithelial cells (cortical and medullary TECs) disrupts the organ-typical thymus vascular architecture, causing hypovascularization, demonstrating that TEC-derived VEGF-A is required for normal thymus vascular morphogenesis.","method":"Conditional gene targeting via nude mouse blastocyst complementation, histological analysis of vascular architecture","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional knockout with defined cellular source and clear structural phenotype, rigorous genetic approach","pmids":["16027358"],"is_preprint":false},{"year":2006,"finding":"VEGF165 protein is cleaved by plasmin at Arg110/Ala111 in the wound microenvironment, reducing its mitogenic activity; mutagenesis of this cleavage site preserves structural integrity and increases angiogenic potency in an impaired healing mouse model. Additionally, soluble VEGFR-1 (sVEGFR-1) acts as an endogenous inhibitor of VEGF-A in non-healing wounds.","method":"Protease cleavage assay, site-directed mutagenesis, in vivo impaired healing mouse model, ELISA for sVEGFR-1","journal":"The journal of investigative dermatology. Symposium proceedings","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro cleavage assay with mutagenesis validated in vivo, single lab","pmids":["17069014"],"is_preprint":false},{"year":2007,"finding":"IFN-γ suppresses monocyte VEGF-A translation via the GAIT (IFN-γ-activated inhibitor of translation) complex, which binds a specific element in the VEGF-A 3′UTR; although IFN-γ induces VEGF-A mRNA, the GAIT complex delays and silences translation, reducing VEGF-A protein and angiogenic activity.","method":"mRNA-protein interaction studies (EMSA/RNA pulldown), translation reporter assays, angiogenesis functional assays","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — direct biochemical identification of GAIT complex binding to VEGF-A 3′UTR, functional translation silencing demonstrated with multiple orthogonal methods, single lab","pmids":["17611605"],"is_preprint":false},{"year":2009,"finding":"HSV-1-induced corneal lymphangiogenesis is strictly dependent on VEGF-A/VEGFR-2 signaling (not VEGFR-3 ligands); infected epithelial cells (not macrophages) are the primary source of VEGF-A during infection, as identified using VEGF-A reporter transgenic mice.","method":"VEGF-A reporter transgenic mice, VEGFR-2 and VEGFR-3 blocking studies, macrophage depletion, in vivo corneal infection model","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple genetic and pharmacological epistasis approaches in vivo, reporter-based cellular source identification, single lab","pmids":["20026662"],"is_preprint":false},{"year":2009,"finding":"VEGF-A and HGF cooperate in endothelial angiogenesis by synergistically activating ERK1/2 and p38 kinases downstream of their respective receptors (VEGFR-2 and c-Met), which do not physically associate or transphosphorylate each other; VEGF-A activates Rho-dependent cytoskeletal remodeling while HGF activates Rac-dependent remodeling.","method":"Co-immunoprecipitation (negative for receptor association), MAPK activation kinetics assays, Rho/Rac GTPase activity assays, in vitro tube formation","journal":"Biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple signaling pathway readouts with negative epistasis for direct receptor interaction, single lab","pmids":["19281453"],"is_preprint":false},{"year":2009,"finding":"Autocrine VEGF-A and VEGF-C in human podocytes both activate anti-apoptotic PI3K/AKT and suppress pro-apoptotic p38MAPK via VEGFR-2; ablation of VEGF-A or VEGF-C, or treatment with bevacizumab or VEGFR-2/-3 inhibitors, reduces podocyte survival. Exogenous VEGF-C can substitute for VEGF-A and vice versa in maintaining survival signaling.","method":"siRNA knockdown, bevacizumab treatment, VEGFR-2/-3 tyrosine kinase inhibition, Western blot for PI3K/AKT and p38MAPK activation, viability assays","journal":"American journal of physiology. Renal physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal siRNA knockdown and pharmacological inhibition with defined signaling readouts, single lab","pmids":["19828679"],"is_preprint":false},{"year":2010,"finding":"VEGF-A expression in osteoclasts is regulated by HIF-1α, which itself is induced downstream of NF-κB activation by RANKL; NF-κB inhibition suppresses HIF-1α mRNA, and HIF-1α inhibition decreases VEGF-A mRNA, placing NF-κB upstream of HIF-1α upstream of VEGF-A in osteoclast signaling.","method":"Specific transcription factor inhibitors (NF-κB, AP-1, NFATc1, HIF-1), RT-PCR, conditioned media VEGF-A secretion assay","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — pharmacological inhibitor epistasis in cell culture with mRNA and protein readouts, single lab","pmids":["20432243"],"is_preprint":false},{"year":2010,"finding":"IRE1α, PERK, and ATF6α of the unfolded protein response (UPR) each regulate VEGF-A mRNA transcription via their respective downstream transcription factors (spliced XBP-1, ATF4, and cleaved ATF6); loss of these UPR components in mouse embryonic fibroblasts or ATF6α-knockdown cells attenuates VEGF-A induction under ER stress.","method":"Genetic knockouts (Ire1α−/−, Perk−/− MEFs), siRNA knockdown (ATF6α), rescue experiments, qRT-PCR","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple genetic loss-of-function models with rescue experiments and transcriptional readout, identifies three parallel UPR pathways","pmids":["20221394"],"is_preprint":false},{"year":2010,"finding":"VEGF-A-induced signaling through VEGFR-2 is concentration- and duration-dependent: PLCγ/ERK1/2 (p44/p42 MAPK) pathway transduces graded VEGF-A concentration information, whereas the parallel AKT pathway does not; longer exposure duration does not compensate for low VEGF-A concentration, indicating these parameters are not equivalent.","method":"Quantitative VEGFR2 autophosphorylation assays, signal kinase activation assays at graded VEGF-A concentrations, immediate-early gene induction analysis","journal":"Microvascular research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — systematic dose-response and time-course signaling experiments in endothelial cells, single lab","pmids":["20144626"],"is_preprint":false},{"year":2011,"finding":"VEGF-A induces degradation of versican in venular basement membranes during mother vessel formation by upregulating ADAMTS-1 (a versican-cleaving protease) and MMP-15 (which activates ADAMTS-1) in endothelial cells.","method":"Immunohistochemistry, Western blot, in vivo adenoviral VEGF-A(164) expression model, in vitro VEGF-A endothelial cell stimulation","journal":"The journal of histochemistry and cytochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro and in vivo corroborating experiments identifying protease induction mechanism, single lab","pmids":["21411713"],"is_preprint":false},{"year":2015,"finding":"VEGF-A produced in the tumor microenvironment enhances expression of PD-1, Tim-3, and other inhibitory checkpoints on CD8+ T cells, promoting T cell exhaustion; this effect can be reverted by anti-angiogenic agents targeting VEGF-A/VEGFR.","method":"In vivo tumor models, flow cytometry for inhibitory receptor expression, anti-VEGF-A/VEGFR treatment reversal experiments","journal":"The Journal of experimental medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo loss-of-function with defined cellular phenotypic readout (checkpoint receptor expression), single lab","pmids":["25601652"],"is_preprint":false},{"year":2015,"finding":"TGF-β1 regulates peritoneal VEGF-A expression through an ID1-dependent pathway in mesothelial cells; siRNA knockdown of ID1 abolishes TGF-β1-induced VEGF-A upregulation, placing ID1 as an intermediary between TGF-β1 and VEGF-A transcription.","method":"siRNA knockdown of ID1, qRT-PCR and ELISA for VEGF-A, TGF-β1 stimulation of primary peritoneal and immortalized mesothelial cells","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA epistasis with protein-level readout, multiple cell types tested, single lab","pmids":["26577912"],"is_preprint":false},{"year":2015,"finding":"VEGF-A165a sensitizes peripheral nociceptive neurons through VEGFR-2 and a TRPV1-dependent mechanism to enhance pain signaling; VEGF-A165b blocks this effect. After nerve injury, SRPK1-dependent pre-mRNA splicing shifts the isoform balance toward VEGF-Axxxa over VEGF-Axxxb, promoting neuropathic pain. Pharmacological inhibition of SRPK1 reverses this shift and alleviates pain.","method":"In vivo pain behavioral assays in rats and mice, VEGFR2 and TRPV1 pharmacological blockade, SRPK1 inhibition, exogenous VEGF-A165b treatment","journal":"Neurobiology of disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo pharmacological epistasis identifying VEGFR2-TRPV1 pathway and SRPK1 splicing regulation, single lab","pmids":["25151644"],"is_preprint":false},{"year":2016,"finding":"Different VEGF-A isoforms (VEGF-A165, VEGF-A121, VEGF-A145) promote distinct patterns of VEGFR-2 endocytosis, differential ubiquitylation, and differential signal transduction; disruption of clathrin-dependent endocytosis blocks isoform-specific VEGFR-2 activation and causes depletion of membrane-bound VEGFR1 and VEGFR2.","method":"Clathrin endocytosis disruption, receptor internalization assays, Western blot for ubiquitylation and signaling, receptor proteolysis assays","journal":"Biology open","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple isoforms compared with receptor trafficking and signaling readouts, clathrin inhibition epistasis, single lab","pmids":["27044325"],"is_preprint":false},{"year":2018,"finding":"VEGF-A isoforms bind VEGFR2 with similar affinities but differ in interactions with co-receptor Neuropilin-1 and heparan sulfate proteoglycans (HSPGs); alternative splicing at exons 5–7 controls heparin-binding and bioavailability, while alternative 3′ splice-site selection at exon 8 generates VEGFxxxb isoforms with distinct downstream signaling.","method":"Receptor binding assays, downstream signaling analysis, VEGF-A isoform pharmacological comparison (review synthesizing experimental data)","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 3 / Strong — review synthesizing experimental results from multiple labs; individual mechanistic findings supported by cited experiments","pmids":["29690653"],"is_preprint":false},{"year":2018,"finding":"Molecular dynamics and circular dichroism modeling reveals that the VEGF-A heparin-binding domain (HBD) forms sandwich-like HBD-heparin-HBD structures that regulate the mutual disposition of HBDs and affect VEGF-A-mediated signaling; conformational flexibility of the 12-amino acid interdomain linker contributes to this regulation.","method":"Molecular docking, molecular dynamics simulation, circular dichroism spectroscopy","journal":"Journal of molecular graphics & modelling","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational modeling with only CD spectroscopy experimental validation, no functional mutagenesis","pmids":["29738889"],"is_preprint":false},{"year":2019,"finding":"TRF2 promotes VEGF-A secretion and angiogenesis by transcriptionally upregulating SULF2 (an endoglucosamine-6-sulfatase) via binding to a distal regulatory element; SULF2 then modifies heparan sulfate proteoglycans to release membrane-associated VEGF-A, increasing its availability.","method":"Luminex multiplexed secretome profiling, TRF2 overexpression/knockdown, ChIP for TRF2 binding, SULF2 knockdown epistasis, endothelial differentiation assay","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP identifies regulatory element, SULF2 epistasis confirms pathway, secretome profiling, single lab","pmids":["30698737"],"is_preprint":false},{"year":2019,"finding":"Granuloma macrophages produce VEGF-A that recruits immune cells to granulomas via a non-angiogenic pathway; selective VEGF-A blockade in myeloid cells or pharmaceutical inhibition reduces granulomatous inflammation and improves survival in Mycobacterium tuberculosis-infected mice without altering host protection.","method":"Conditional myeloid-specific VEGF-A knockout, granuloma transplantation, pharmaceutical VEGF-A inhibition, in vivo Mtb and BCG infection models","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional genetic knockout combined with pharmacological inhibition and transplantation epistasis in multiple infection models","pmids":["31091450"],"is_preprint":false},{"year":2021,"finding":"Endothelial insulin receptors are required for appropriate VEGF-A signal transduction from VEGFR-2 to ERK1/2; insulin receptor haploinsufficiency impairs VEGFR-2 internalization (required for ERK1/2 signaling) and abolishes VEGF-A-induced ERK1/2 activation, while VEGF-A signaling to Akt and eNOS remains intact.","method":"Whole-body and endothelium-restricted insulin receptor haploinsufficient mice, shRNA-mediated Insr knockdown in HUVECs, Western blot for ERK1/2 and Akt, VEGFR-2 internalization assays, in vitro and in vivo angiogenesis assays","journal":"Endocrinology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple genetic models (whole-body and endothelium-restricted KO) plus shRNA knockdown with defined signaling pathway dissection, two orthogonal experimental systems","pmids":["34037749"],"is_preprint":false},{"year":2004,"finding":"Laminin and α3β1 integrin signaling regulates constitutive VEGF-A expression in podocytes via a non-hypoxic HIF-α-dependent mechanism; classical PKC is a potential intermediary; HIF-α activity in podocytes is increased independently of hypoxia and drives VEGF-A promoter activity.","method":"VEGF-A promoter-luciferase reporter assay, quantitative RT-PCR, ELISA, immunoprecipitation for HIF-α/p300 association, extracellular matrix stimulation","journal":"Kidney international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reporter assay with matrix-specific conditions and HIF-α co-immunoprecipitation, single lab preliminary study","pmids":["15458440"],"is_preprint":false},{"year":2014,"finding":"VEGF-A-regulated gene transcription in endothelial cells is controlled at two levels: RNA polymerase II pausing at promoters and productive elongation; approximately half of VEGF-A-regulated promoters show paused Pol II, and transition to productive elongation is a major activation mechanism for virtually all VEGF-regulated genes.","method":"Genome-wide GRO-Seq (global run-on sequencing), tethered conformation capture (TCC) chromatin interaction mapping in primary HAECs and HUVECs","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1 / Moderate — genome-wide mechanistic transcriptional assay (GRO-Seq) with chromatin interaction mapping, two primary cell types, single lab","pmids":["25352550"],"is_preprint":false},{"year":2015,"finding":"WISP-1 promotes VEGF-A expression in osteosarcoma cells and angiogenesis through activation of the FAK/JNK/HIF-1α signaling pathway and down-regulation of miR-381; inhibitors of FAK, JNK, or HIF-1α, or HIF-1α siRNA, abolish WISP-1-induced VEGF-A expression.","method":"Pathway inhibitors (FAK, JNK, HIF-1α), siRNA knockdown, in vitro EPC migration/tube formation, in vivo tumor xenograft, immunohistochemistry","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological and genetic epistasis with in vitro and in vivo validation, single lab","pmids":["28406476"],"is_preprint":false},{"year":2021,"finding":"VEGF-A can signal intracellularly (intracrine signaling) within cells expressing both VEGF-A and its receptors, regulating cell growth, survival, and metabolism independently of secretion; this intracrine mode involves VEGFR1 and VEGFR2 expressed intracellularly.","method":"Review synthesizing loss-of-function and localization experiments from multiple studies demonstrating intracellular VEGF-A/receptor complexes","journal":"Biomolecules","confidence":"Low","confidence_rationale":"Tier 3 / Weak — review synthesizing others' findings; individual mechanistic experiments not described in the abstract itself","pmids":["33478167"],"is_preprint":false},{"year":2012,"finding":"VEGF-A expressed by pigmented ciliary epithelium maintains ciliary body homeostasis; systemic VEGF-A neutralization (via adenoviral sFlt1 overexpression) causes thinning of nonpigmented epithelium, vacuolization of pigmented epithelium, loss of capillary fenestrations, thrombosis, and decreased intraocular pressure, indicating a required role in ciliary body vascular and epithelial maintenance.","method":"VEGF-A neutralization via adenoviral sFlt1, VEGF-LacZ reporter mice, VEGFR2 localization by immunostaining, light microscopy and TEM, intraocular pressure measurement","journal":"Investigative ophthalmology & visual science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo VEGF-A neutralization with defined structural and functional phenotypic readouts, reporter-based localization, single lab","pmids":["23081980"],"is_preprint":false},{"year":2020,"finding":"In multiple myeloma, FGF23 produced by osteocytes in response to direct contact with myeloma cells upregulates VEGF-A expression in osteocytes; VEGF-A knockdown in osteocytes or VEGF-A neutralization in conditioned media completely abolishes the increased endothelial tube formation induced by myeloma–osteocyte co-culture.","method":"siRNA knockdown of Vegf-a in osteoclasts/primary osteocytes, VEGF-A neutralization, co-culture endothelial tube formation assay, Fgf23 deletion, in vivo myeloma mouse model with vessel area quantification","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockdown and neutralization epistasis in vitro and in vivo with mechanistic Fgf23–Vegf-a pathway link, single lab","pmids":["33057033"],"is_preprint":false}],"current_model":"VEGF-A is a secreted dimeric glycoprotein that acts primarily through two high-affinity tyrosine kinase receptors (VEGFR-1 and VEGFR-2) to activate endothelial cells via PLCγ/ERK1/2, PI3K/Akt/eNOS, and other downstream pathways, driving angiogenesis, vascular permeability, lymphangiogenesis, and immune cell modulation; multiple alternatively spliced isoforms differ in heparin-binding, bioavailability, Neuropilin-1 co-receptor interaction, and VEGFR-2 trafficking/signaling outputs; VEGF-A translation is post-transcriptionally silenced by the IFN-γ–induced GAIT complex binding to its 3′UTR; its transcription is regulated by HIF-1α (under hypoxia and non-hypoxically via integrin signaling in podocytes), UPR sensors (IRE1α-XBP-1, PERK-ATF4, ATF6α), NF-κB, and chromatin-level Pol II pausing/elongation control; extracellular VEGF-A bioavailability is further regulated by plasmin cleavage, SULF2-mediated heparan sulfate modification, and soluble VEGFR-1 sequestration; VEGF-A also drives non-angiogenic functions including T-cell exhaustion via PD-1/Tim-3 upregulation, immune-cell recruitment to granulomas, nociceptor sensitization via VEGFR-2/TRPV1, and autocrine podocyte survival via PI3K/AKT/p38MAPK."},"narrative":{"mechanistic_narrative":"VEGF-A is a secreted, disulfide-bonded dimeric heparin-binding glycoprotein that signals through two high-affinity endothelial tyrosine kinase receptors (VEGFR-1/Flt-1 and VEGFR-2) to drive angiogenesis, vascular permeability, lymphangiogenesis, and tissue-specific vascular maintenance [PMID:8281615, PMID:16027358, PMID:23081980]. Receptor engagement activates parallel downstream cascades whose outputs are non-equivalent: VEGFR-2-coupled PLCγ/ERK1/2 signaling transduces graded VEGF-A concentration information, whereas the Akt arm operates as a concentration-independent switch that is both necessary and sufficient for vascular permeability through eNOS [PMID:12459464, PMID:20144626]. Efficient ERK1/2 activation requires VEGFR-2 internalization, which itself depends on the endothelial insulin receptor, while signaling to Akt/eNOS proceeds independently [PMID:34037749]. Beyond endothelium, VEGF-A drives mural-cell recruitment by inducing endothelial PDGF-B [PMID:16105884], promotes chemotaxis of mesenchymal progenitors via VEGFR-1 [PMID:16005848], remodels the venular basement membrane by upregulating ADAMTS-1 and MMP-15 [PMID:21411713], and supports autocrine podocyte survival via VEGFR-2-driven PI3K/AKT with suppression of p38MAPK [PMID:19828679]. VEGF-A also executes non-angiogenic programs: it sensitizes nociceptors through a VEGFR-2/TRPV1 mechanism [PMID:25151644], promotes CD8+ T-cell exhaustion by upregulating PD-1 and Tim-3 [PMID:25601652], and recruits immune cells to tuberculous granulomas from a myeloid source [PMID:31091450]. Its output is shaped extensively by alternative splicing—exon 5–7 selection tunes heparin/HSPG and Neuropilin-1 binding and bioavailability, while exon 8 3′-splice-site choice generates VEGFxxxb isoforms—and distinct isoforms drive distinct VEGFR-2 endocytosis, ubiquitylation, and signaling [PMID:27044325, PMID:29690653, PMID:25151644]. Transcription is governed by HIF-1α (under hypoxia and non-hypoxically via integrin signaling), NF-κB-to-HIF-1α cascades, the UPR sensors IRE1α-XBP-1/PERK-ATF4/ATF6α, and Pol II pausing-to-elongation control [PMID:20432243, PMID:20221394, PMID:15458440, PMID:25352550], while translation is silenced post-transcriptionally by the IFN-γ-induced GAIT complex binding the 3′UTR [PMID:17611605]. Extracellular bioavailability is further set by plasmin cleavage at Arg110/Ala111, soluble VEGFR-1 sequestration, and SULF2-mediated heparan sulfate modification that releases matrix-bound ligand [PMID:17069014, PMID:30698737].","teleology":[{"year":1993,"claim":"Established VEGF-A as a secreted dimeric heparin-binding glycoprotein that acts directly on endothelial cells through two high-affinity tyrosine kinase receptors, defining the basic receptor-ligand axis.","evidence":"Receptor binding and functional in vitro assays (PLC activation, calcium transients), biochemical characterization","pmids":["8281615"],"confidence":"High","gaps":["Did not resolve which receptor mediates which downstream output","No isoform-specific signaling distinguished"]},{"year":2002,"claim":"Showed that the Akt arm is both necessary and sufficient for VEGF-A-induced vascular permeability and acts through eNOS, separating permeability control from other VEGF outputs.","evidence":"Adenoviral dominant-negative/constitutively active Akt in vivo Miles assay with eNOS inhibitor blockade","pmids":["12459464"],"confidence":"High","gaps":["Did not address how Akt is selectively activated downstream of which receptor","Relationship to ERK1/2 arm not defined"]},{"year":2004,"claim":"Demonstrated non-hypoxic transcriptional control of VEGF-A by laminin/α3β1 integrin signaling acting through HIF-α in podocytes, expanding HIF regulation beyond oxygen sensing.","evidence":"VEGF-A promoter-luciferase reporter, HIF-α/p300 co-IP, matrix stimulation in podocytes","pmids":["15458440"],"confidence":"Medium","gaps":["PKC intermediary only a candidate","Mechanism of non-hypoxic HIF-α stabilization not resolved"]},{"year":2005,"claim":"Defined cell-type-specific developmental and pathological angiogenic roles for VEGF-A: TEC-derived VEGF-A for thymus vascular morphogenesis, mural-cell recruitment via induced PDGF-B, VEGFR-1-mediated progenitor chemotaxis, and tumor-driven sentinel-node lymphangiogenesis.","evidence":"Conditional gene targeting, Matrigel plug and neutralizing antibody epistasis, Boyden chamber chemotaxis, VEGF-A transgenic mice with lymphatic IHC","pmids":["16027358","16105884","16005848","15809353"],"confidence":"High","gaps":["Receptor selectivity for lymphangiogenesis vs angiogenesis only partly resolved","Crosstalk between PDGF-B and direct VEGF effects not fully separated"]},{"year":2006,"claim":"Identified extracellular regulation of VEGF-A activity by plasmin cleavage at Arg110/Ala111 and soluble VEGFR-1 sequestration, establishing post-secretion control of angiogenic potency.","evidence":"Protease cleavage assay, site-directed mutagenesis, impaired-healing mouse model, sVEGFR-1 ELISA","pmids":["17069014"],"confidence":"Medium","gaps":["In vivo contribution of cleavage vs sVEGFR-1 not quantitatively partitioned","Other proteases not excluded"]},{"year":2007,"claim":"Revealed post-transcriptional silencing of VEGF-A translation by the IFN-γ-induced GAIT complex binding its 3′UTR, decoupling mRNA induction from protein output.","evidence":"RNA-protein interaction (EMSA/pulldown), translation reporter and angiogenesis functional assays","pmids":["17611605"],"confidence":"High","gaps":["GAIT regulation shown in monocytes; generality across cell types not established","Kinetics of silencing onset not fully defined"]},{"year":2009,"claim":"Mapped non-angiogenic and cooperative signaling roles: autocrine podocyte survival via VEGFR-2/PI3K-AKT/p38, VEGF-A/HGF synergy via independent receptors with distinct Rho vs Rac remodeling, and VEGF-A/VEGFR-2 as the obligate driver of infection-induced corneal lymphangiogenesis.","evidence":"siRNA and pharmacological inhibition with signaling readouts, co-IP (negative for receptor association), Rho/Rac assays, VEGF-A reporter mice and receptor blockade in corneal infection","pmids":["19828679","19281453","20026662"],"confidence":"Medium","gaps":["VEGF-A vs VEGF-C redundancy in podocytes not fully separated","Synergy mechanism limited to MAPK readouts"]},{"year":2010,"claim":"Resolved upstream transcriptional logic (NF-κB→HIF-1α, three parallel UPR arms) and demonstrated that VEGFR-2 PLCγ/ERK1/2 encodes graded ligand concentration whereas Akt does not.","evidence":"Transcription factor inhibitor epistasis, Ire1α/Perk knockout MEFs and ATF6α knockdown with rescue, quantitative dose-response/time-course VEGFR-2 signaling assays","pmids":["20432243","20221394","20144626"],"confidence":"High","gaps":["How concentration is decoded into distinct gene programs unresolved","Integration of multiple UPR arms not quantified"]},{"year":2011,"claim":"Showed VEGF-A drives basement-membrane remodeling during mother vessel formation by inducing endothelial ADAMTS-1 and its activator MMP-15 to degrade versican.","evidence":"IHC, Western blot, in vivo adenoviral VEGF-A164 and in vitro endothelial stimulation","pmids":["21411713"],"confidence":"Medium","gaps":["Direct functional requirement of versican cleavage for vessel formation not tested","Single lab"]},{"year":2014,"claim":"Established that VEGF-A-induced endothelial transcription is controlled by RNA Pol II pausing and transition to productive elongation across most regulated genes.","evidence":"Genome-wide GRO-Seq and tethered conformation capture in primary HAECs/HUVECs","pmids":["25352550"],"confidence":"High","gaps":["Pause-release factors recruited by VEGF signaling not identified","Link to specific upstream kinase arms unmapped"]},{"year":2015,"claim":"Extended VEGF-A function to immune suppression (CD8+ T-cell exhaustion via PD-1/Tim-3), nociceptor sensitization (VEGFR-2/TRPV1 with SRPK1-controlled isoform balance), and added ID1 as a TGF-β1→VEGF-A transcriptional intermediary.","evidence":"In vivo tumor models with checkpoint flow cytometry, pain behavioral assays with VEGFR2/TRPV1/SRPK1 manipulation, ID1 siRNA epistasis","pmids":["25601652","25151644","26577912"],"confidence":"Medium","gaps":["Direct vs indirect VEGF-A action on T cells not fully resolved","Tissue specificity of ID1 pathway unknown"]},{"year":2016,"claim":"Demonstrated isoform-specific control of VEGFR-2 endocytosis, ubiquitylation, and signaling, with clathrin-dependent internalization required for isoform-selective receptor activation.","evidence":"Clathrin disruption, receptor internalization and ubiquitylation assays, isoform comparison","pmids":["27044325"],"confidence":"Medium","gaps":["Structural basis of differential trafficking not defined","Downstream transcriptional consequences not mapped"]},{"year":2018,"claim":"Consolidated the splicing-to-function logic: exon 5–7 selection tunes heparin/HSPG and Neuropilin-1 binding and bioavailability, exon 8 generates VEGFxxxb isoforms, and HBD-heparin-HBD architecture modulates signaling.","evidence":"Review of receptor binding and signaling data; molecular dynamics and circular dichroism of the heparin-binding domain","pmids":["29690653","29738889"],"confidence":"Medium","gaps":["HBD conformational model rests on computation with limited experimental validation","Functional mutagenesis of HBD disposition lacking"]},{"year":2019,"claim":"Identified upstream bioavailability and inflammatory controls: TRF2-driven SULF2 transcription releases matrix-bound VEGF-A, and myeloid-derived VEGF-A recruits immune cells to tuberculous granulomas non-angiogenically.","evidence":"ChIP and SULF2 epistasis with secretome profiling; myeloid-specific VEGF-A knockout, granuloma transplantation, and pharmacological inhibition in Mtb/BCG infection","pmids":["30698737","31091450"],"confidence":"High","gaps":["Whether HS remodeling generalizes beyond endothelial differentiation untested","Immune-cell types recruited and receptor used in granulomas not fully defined"]},{"year":2020,"claim":"Defined a niche signaling circuit in which myeloma-induced osteocyte FGF23 upregulates osteocyte VEGF-A to drive endothelial tube formation.","evidence":"Vegf-a siRNA and neutralization, co-culture tube formation, Fgf23 deletion, in vivo myeloma model","pmids":["33057033"],"confidence":"Medium","gaps":["Receptor mediating FGF23→VEGF-A in osteocytes not identified","Generality beyond myeloma niche unknown"]},{"year":2021,"claim":"Demonstrated that endothelial insulin receptor is required for VEGFR-2 internalization and ERK1/2 signaling selectively, while Akt/eNOS signaling remains intact, linking metabolic receptor status to angiogenic signal routing; intracrine VEGF-A signaling was also proposed.","evidence":"Whole-body and endothelium-restricted Insr haploinsufficient mice plus HUVEC shRNA with signaling and internalization readouts; review synthesis for intracrine mode","pmids":["34037749","33478167"],"confidence":"High","gaps":["Molecular mechanism linking insulin receptor to VEGFR-2 endocytosis unresolved","Intracrine model rests on review-level synthesis without primary mechanistic experiments here"]},{"year":null,"claim":"How the multiple parallel transcriptional inputs, splicing decisions, receptor trafficking states, and extracellular bioavailability controls are integrated to produce a specific VEGF-A signaling output in a given cell remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model linking isoform identity to trafficking to transcriptional program","Quantitative contribution of each bioavailability control in vivo unknown","Intracrine signaling not mechanistically established in primary data"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[0,3,13,18]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,1,13]}],"localization":[{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[0,6,21]},{"term_id":"GO:0031012","term_label":"extracellular matrix","supporting_discovery_ids":[14,21]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,13,23]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[2,4,5,14]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[15,22]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[11,12,24,25]},{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[7,17]}],"complexes":[],"partners":["FLT1","KDR","NRP1","FLT4"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P15692","full_name":"Vascular endothelial growth factor A, long form","aliases":["Vascular permeability factor","VPF"],"length_aa":395,"mass_kda":43.6,"function":"Participates in the induction of key genes involved in the response to hypoxia and in the induction of angiogenesis such as HIF1A (PubMed:35455969). Involved in protecting cells from hypoxia-mediated cell death (By similarity) Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth (PubMed:34530889). Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. Binds to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin. Binds to the NRP1/neuropilin-1 receptor. Binding to NRP1 initiates a signaling pathway needed for motor neuron axon guidance and cell body migration, including for the caudal migration of facial motor neurons from rhombomere 4 to rhombomere 6 during embryonic development (By similarity). Also binds the DEAR/FBXW7-AS1 receptor (PubMed:17446437) Binds to the KDR receptor but does not activate downstream signaling pathways, does not activate angiogenesis and inhibits tumor growth","subcellular_location":"Secreted","url":"https://www.uniprot.org/uniprotkb/P15692/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/VEGFA","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/VEGFA","total_profiled":1310},"omim":[{"mim_id":"621467","title":"NCBP2 ANTISENSE RNA 2 (HEAD TO HEAD); NCBP2AS2","url":"https://www.omim.org/entry/621467"},{"mim_id":"621032","title":"CEREBRAL CAVERNOUS MALFORMATIONS 5; CCM5","url":"https://www.omim.org/entry/621032"},{"mim_id":"620997","title":"SEMAPHORIN 3G; SEMA3G","url":"https://www.omim.org/entry/620997"},{"mim_id":"620529","title":"RING FINGER PROTEIN 121; RNF121","url":"https://www.omim.org/entry/620529"},{"mim_id":"620215","title":"MEMBRANE INTEGRAL NOTCH2-ASSOCIATED RECEPTOR 2; MINAR2","url":"https://www.omim.org/entry/620215"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/VEGFA"},"hgnc":{"alias_symbol":["VEGF-A","VPF"],"prev_symbol":["VEGF"]},"alphafold":{"accession":"P15692","domains":[{"cath_id":"2.10.90.10","chopping":"42-131","consensus_level":"high","plddt":95.7278,"start":42,"end":131},{"cath_id":"2.10.160.10","chopping":"194-227","consensus_level":"high","plddt":83.6382,"start":194,"end":227}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P15692","model_url":"https://alphafold.ebi.ac.uk/files/AF-P15692-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P15692-F1-predicted_aligned_error_v6.png","plddt_mean":63.91},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=VEGFA","jax_strain_url":"https://www.jax.org/strain/search?query=VEGFA"},"sequence":{"accession":"P15692","fasta_url":"https://rest.uniprot.org/uniprotkb/P15692.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P15692/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P15692"}},"corpus_meta":[{"pmid":"25601652","id":"PMC_25601652","title":"VEGF-A modulates expression of inhibitory checkpoints on CD8+ T cells in tumors.","date":"2015","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/25601652","citation_count":964,"is_preprint":false},{"pmid":"8281615","id":"PMC_8281615","title":"Vascular permeability factor (VPF, VEGF) in tumor biology.","date":"1993","source":"Cancer metastasis reviews","url":"https://pubmed.ncbi.nlm.nih.gov/8281615","citation_count":788,"is_preprint":false},{"pmid":"15809353","id":"PMC_15809353","title":"VEGF-A induces tumor and sentinel lymph node lymphangiogenesis and promotes lymphatic metastasis.","date":"2005","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/15809353","citation_count":598,"is_preprint":false},{"pmid":"29690653","id":"PMC_29690653","title":"Molecular Pharmacology of VEGF-A Isoforms: Binding and Signalling at VEGFR2.","date":"2018","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/29690653","citation_count":389,"is_preprint":false},{"pmid":"19013462","id":"PMC_19013462","title":"VEGF-A links angiogenesis and inflammation in inflammatory bowel disease pathogenesis.","date":"2008","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/19013462","citation_count":289,"is_preprint":false},{"pmid":"22434866","id":"PMC_22434866","title":"Diverse roles for VEGF-A in the nervous system.","date":"2012","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/22434866","citation_count":231,"is_preprint":false},{"pmid":"16105884","id":"PMC_16105884","title":"VEGF-A and FGF-2 synergistically promote neoangiogenesis through enhancement of endogenous PDGF-B-PDGFRbeta signaling.","date":"2005","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/16105884","citation_count":230,"is_preprint":false},{"pmid":"20221394","id":"PMC_20221394","title":"Transcriptional regulation of VEGF-A by the unfolded protein response pathway.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/20221394","citation_count":225,"is_preprint":false},{"pmid":"17475821","id":"PMC_17475821","title":"Interstitial vascular rarefaction and reduced VEGF-A expression in human diabetic nephropathy.","date":"2007","source":"Journal of the American Society of Nephrology : JASN","url":"https://pubmed.ncbi.nlm.nih.gov/17475821","citation_count":207,"is_preprint":false},{"pmid":"11687953","id":"PMC_11687953","title":"VEGF-A, VEGF-C, and VEGF-D in colorectal cancer progression.","date":"2001","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/11687953","citation_count":178,"is_preprint":false},{"pmid":"32722551","id":"PMC_32722551","title":"VEGF-A in Cardiomyocytes and Heart Diseases.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32722551","citation_count":176,"is_preprint":false},{"pmid":"15090854","id":"PMC_15090854","title":"The role of VEGF-A in glomerular development and function.","date":"2004","source":"Current opinion in nephrology and hypertension","url":"https://pubmed.ncbi.nlm.nih.gov/15090854","citation_count":158,"is_preprint":false},{"pmid":"20026662","id":"PMC_20026662","title":"VEGF-A expression by HSV-1-infected cells drives corneal lymphangiogenesis.","date":"2009","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/20026662","citation_count":153,"is_preprint":false},{"pmid":"16005848","id":"PMC_16005848","title":"VEGF-A and PlGF-1 stimulate chemotactic migration of human mesenchymal progenitor cells.","date":"2005","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/16005848","citation_count":153,"is_preprint":false},{"pmid":"33854498","id":"PMC_33854498","title":"Direct and Indirect Modulation of T Cells by VEGF-A Counteracted by Anti-Angiogenic Treatment.","date":"2021","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/33854498","citation_count":151,"is_preprint":false},{"pmid":"19281453","id":"PMC_19281453","title":"Cross-talk between the VEGF-A and HGF signalling pathways in endothelial cells.","date":"2009","source":"Biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/19281453","citation_count":139,"is_preprint":false},{"pmid":"28752859","id":"PMC_28752859","title":"MicroRNA-140-5p inhibits invasion and angiogenesis through targeting VEGF-A in breast cancer.","date":"2017","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/28752859","citation_count":136,"is_preprint":false},{"pmid":"17264876","id":"PMC_17264876","title":"Reduction of VEGF-A and CTGF expression in diabetic nephropathy is associated with podocyte loss.","date":"2007","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/17264876","citation_count":135,"is_preprint":false},{"pmid":"24944907","id":"PMC_24944907","title":"Brown adipose tissue derived VEGF-A modulates cold tolerance and energy expenditure.","date":"2014","source":"Molecular metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/24944907","citation_count":130,"is_preprint":false},{"pmid":"10709865","id":"PMC_10709865","title":"VPF/VEGF and the angiogenic response.","date":"2000","source":"Seminars in perinatology","url":"https://pubmed.ncbi.nlm.nih.gov/10709865","citation_count":125,"is_preprint":false},{"pmid":"19521900","id":"PMC_19521900","title":"Molecular diversity of VEGF-A as a regulator of its biological activity.","date":"2009","source":"Microcirculation (New York, N.Y. : 1994)","url":"https://pubmed.ncbi.nlm.nih.gov/19521900","citation_count":113,"is_preprint":false},{"pmid":"14703061","id":"PMC_14703061","title":"Tumor specific VEGF-A and VEGFR2/KDR protein are co-expressed in breast cancer.","date":"2003","source":"Breast cancer research and treatment","url":"https://pubmed.ncbi.nlm.nih.gov/14703061","citation_count":104,"is_preprint":false},{"pmid":"17069014","id":"PMC_17069014","title":"Molecular mechanisms of VEGF-A action during tissue repair.","date":"2006","source":"The journal of investigative dermatology. Symposium proceedings","url":"https://pubmed.ncbi.nlm.nih.gov/17069014","citation_count":98,"is_preprint":false},{"pmid":"17611605","id":"PMC_17611605","title":"A post-transcriptional pathway represses monocyte VEGF-A expression and angiogenic activity.","date":"2007","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/17611605","citation_count":93,"is_preprint":false},{"pmid":"30504800","id":"PMC_30504800","title":"Modified VEGF-A mRNA induces sustained multifaceted microvascular response and accelerates diabetic wound healing.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/30504800","citation_count":91,"is_preprint":false},{"pmid":"26745345","id":"PMC_26745345","title":"A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo.","date":"2016","source":"ACS chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/26745345","citation_count":88,"is_preprint":false},{"pmid":"33469143","id":"PMC_33469143","title":"BMP-2 and VEGF-A modRNAs in collagen scaffold synergistically drive bone repair through osteogenic and angiogenic pathways.","date":"2021","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/33469143","citation_count":81,"is_preprint":false},{"pmid":"15661835","id":"PMC_15661835","title":"VEGF-A splice variants and related receptor expression in human skeletal muscle following submaximal exercise.","date":"2005","source":"Journal of applied physiology (Bethesda, Md. : 1985)","url":"https://pubmed.ncbi.nlm.nih.gov/15661835","citation_count":75,"is_preprint":false},{"pmid":"33478167","id":"PMC_33478167","title":"Exploring the Intracrine Functions of VEGF-A.","date":"2021","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/33478167","citation_count":73,"is_preprint":false},{"pmid":"25151644","id":"PMC_25151644","title":"Regulation of alternative VEGF-A mRNA splicing is a therapeutic target for analgesia.","date":"2014","source":"Neurobiology of disease","url":"https://pubmed.ncbi.nlm.nih.gov/25151644","citation_count":71,"is_preprint":false},{"pmid":"28406476","id":"PMC_28406476","title":"WISP-1 positively regulates angiogenesis by controlling VEGF-A expression in human osteosarcoma.","date":"2017","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/28406476","citation_count":68,"is_preprint":false},{"pmid":"30510170","id":"PMC_30510170","title":"Clostridium difficile toxins induce VEGF-A and vascular permeability to promote disease pathogenesis.","date":"2018","source":"Nature microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/30510170","citation_count":66,"is_preprint":false},{"pmid":"26373346","id":"PMC_26373346","title":"Nanoparticle Delivered VEGF-A siRNA Enhances Photodynamic Therapy for Head and Neck Cancer Treatment.","date":"2015","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/26373346","citation_count":65,"is_preprint":false},{"pmid":"12692089","id":"PMC_12692089","title":"Adenovirus-mediated VEGF-A gene transfer induces bone formation in vivo.","date":"2003","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/12692089","citation_count":64,"is_preprint":false},{"pmid":"25697344","id":"PMC_25697344","title":"Eicosapentaenoic acid upregulates VEGF-A through both GPR120 and PPARγ mediated pathways in 3T3-L1 adipocytes.","date":"2015","source":"Molecular and cellular endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/25697344","citation_count":60,"is_preprint":false},{"pmid":"20042655","id":"PMC_20042655","title":"Regulation of VEGF-A in uveal melanoma.","date":"2009","source":"Investigative ophthalmology & visual science","url":"https://pubmed.ncbi.nlm.nih.gov/20042655","citation_count":59,"is_preprint":false},{"pmid":"31414144","id":"PMC_31414144","title":"VEGF-A and blood vessels: a beta cell perspective.","date":"2019","source":"Diabetologia","url":"https://pubmed.ncbi.nlm.nih.gov/31414144","citation_count":58,"is_preprint":false},{"pmid":"21194429","id":"PMC_21194429","title":"VEGF₁₂₁b and VEGF₁₆₅b are weakly angiogenic isoforms of VEGF-A.","date":"2010","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/21194429","citation_count":58,"is_preprint":false},{"pmid":"19828679","id":"PMC_19828679","title":"The balance of autocrine VEGF-A and VEGF-C determines podocyte survival.","date":"2009","source":"American journal of physiology. Renal physiology","url":"https://pubmed.ncbi.nlm.nih.gov/19828679","citation_count":57,"is_preprint":false},{"pmid":"23260712","id":"PMC_23260712","title":"VEGF-C, VEGF-A and related angiogenesis factors as biomarkers of allograft vasculopathy in cardiac transplant recipients.","date":"2013","source":"The Journal of heart and lung transplantation : the official publication of the International Society for Heart Transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/23260712","citation_count":57,"is_preprint":false},{"pmid":"35303290","id":"PMC_35303290","title":"How VEGF-A and its splice variants affect breast cancer development - clinical implications.","date":"2022","source":"Cellular oncology (Dordrecht, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/35303290","citation_count":56,"is_preprint":false},{"pmid":"16027358","id":"PMC_16027358","title":"Gene targeting of VEGF-A in thymus epithelium disrupts thymus blood vessel architecture.","date":"2005","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/16027358","citation_count":56,"is_preprint":false},{"pmid":"12459464","id":"PMC_12459464","title":"Akt signaling mediates VEGF/VPF vascular permeability in vivo.","date":"2002","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/12459464","citation_count":55,"is_preprint":false},{"pmid":"26577912","id":"PMC_26577912","title":"Peritoneal VEGF-A expression is regulated by TGF-β1 through an ID1 pathway in women with endometriosis.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26577912","citation_count":53,"is_preprint":false},{"pmid":"18240146","id":"PMC_18240146","title":"Specific imaging of VEGF-A expression with radiolabeled anti-VEGF monoclonal antibody.","date":"2008","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/18240146","citation_count":53,"is_preprint":false},{"pmid":"38687357","id":"PMC_38687357","title":"Regulation of VEGF-A expression and VEGF-A-targeted therapy in malignant tumors.","date":"2024","source":"Journal of cancer research and clinical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/38687357","citation_count":52,"is_preprint":false},{"pmid":"31091450","id":"PMC_31091450","title":"VEGF-A from Granuloma Macrophages Regulates Granulomatous Inflammation by a Non-angiogenic Pathway during Mycobacterial Infection.","date":"2019","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/31091450","citation_count":51,"is_preprint":false},{"pmid":"21411713","id":"PMC_21411713","title":"Proteolytic cleavage of versican and involvement of ADAMTS-1 in VEGF-A/VPF-induced pathological angiogenesis.","date":"2011","source":"The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society","url":"https://pubmed.ncbi.nlm.nih.gov/21411713","citation_count":51,"is_preprint":false},{"pmid":"36586634","id":"PMC_36586634","title":"Efficient delivery of VEGF-A mRNA for promoting diabetic wound healing via ionizable lipid nanoparticles.","date":"2022","source":"International journal of pharmaceutics","url":"https://pubmed.ncbi.nlm.nih.gov/36586634","citation_count":50,"is_preprint":false},{"pmid":"27645229","id":"PMC_27645229","title":"Expressions of VEGF-A and VEGFR-2 in placentae from GDM pregnancies.","date":"2016","source":"Reproductive biology and endocrinology : RB&E","url":"https://pubmed.ncbi.nlm.nih.gov/27645229","citation_count":49,"is_preprint":false},{"pmid":"27044325","id":"PMC_27044325","title":"VEGF-A isoforms program differential VEGFR2 signal transduction, trafficking and proteolysis.","date":"2016","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/27044325","citation_count":48,"is_preprint":false},{"pmid":"30698737","id":"PMC_30698737","title":"TRF2 positively regulates SULF2 expression increasing VEGF-A release and activity in tumor microenvironment.","date":"2019","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/30698737","citation_count":45,"is_preprint":false},{"pmid":"20432243","id":"PMC_20432243","title":"VEGF-A expression in osteoclasts is regulated by NF-kappaB induction of HIF-1alpha.","date":"2010","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20432243","citation_count":44,"is_preprint":false},{"pmid":"15475575","id":"PMC_15475575","title":"Regulation of the endogenous VEGF-A gene by exogenous designed regulatory proteins.","date":"2004","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/15475575","citation_count":43,"is_preprint":false},{"pmid":"12084990","id":"PMC_12084990","title":"VEGF-A and -C but not -B mediate increased vascular permeability in preserved lung grafts.","date":"2002","source":"Transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/12084990","citation_count":43,"is_preprint":false},{"pmid":"39831819","id":"PMC_39831819","title":"Extracellular vesicle-mediated VEGF-A mRNA delivery rescues ischaemic injury with low immunogenicity.","date":"2025","source":"European heart journal","url":"https://pubmed.ncbi.nlm.nih.gov/39831819","citation_count":42,"is_preprint":false},{"pmid":"20179357","id":"PMC_20179357","title":"Role of VEGF-A in pancreatic beta cells.","date":"2010","source":"Endocrine journal","url":"https://pubmed.ncbi.nlm.nih.gov/20179357","citation_count":42,"is_preprint":false},{"pmid":"24845798","id":"PMC_24845798","title":"Expression profiling and significance of VEGF-A, VEGFR2, VEGFR3 and related proteins in endometrial carcinoma.","date":"2014","source":"Cytokine","url":"https://pubmed.ncbi.nlm.nih.gov/24845798","citation_count":41,"is_preprint":false},{"pmid":"24186558","id":"PMC_24186558","title":"Cooperative effects of FGF-2 and VEGF-A in periodontal ligament cells.","date":"2013","source":"Journal of dental research","url":"https://pubmed.ncbi.nlm.nih.gov/24186558","citation_count":40,"is_preprint":false},{"pmid":"25352550","id":"PMC_25352550","title":"Control of VEGF-A transcriptional programs by pausing and genomic compartmentalization.","date":"2014","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/25352550","citation_count":40,"is_preprint":false},{"pmid":"22547925","id":"PMC_22547925","title":"PEDF and VEGF-A output from human retinal pigment epithelial cells grown on novel microcarriers.","date":"2012","source":"Journal of biomedicine & biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/22547925","citation_count":40,"is_preprint":false},{"pmid":"32573828","id":"PMC_32573828","title":"LPS-mediated neutrophil VEGF-A release is modulated by cannabinoid receptor activation.","date":"2020","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/32573828","citation_count":39,"is_preprint":false},{"pmid":"35749494","id":"PMC_35749494","title":"Human organ rejuvenation by VEGF-A: Lessons from the skin.","date":"2022","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/35749494","citation_count":39,"is_preprint":false},{"pmid":"32533103","id":"PMC_32533103","title":"miR-503-5p inhibits colon cancer tumorigenesis, angiogenesis, and lymphangiogenesis by directly downregulating VEGF-A.","date":"2020","source":"Gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/32533103","citation_count":39,"is_preprint":false},{"pmid":"23651727","id":"PMC_23651727","title":"Intracranial meningiomas, the VEGF-A pathway, and peritumoral brain oedema.","date":"2013","source":"Danish medical journal","url":"https://pubmed.ncbi.nlm.nih.gov/23651727","citation_count":38,"is_preprint":false},{"pmid":"39708087","id":"PMC_39708087","title":"Emerging clinical evidence of a dual role for Ang-2 and VEGF-A blockade with faricimab in retinal diseases.","date":"2024","source":"Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie","url":"https://pubmed.ncbi.nlm.nih.gov/39708087","citation_count":36,"is_preprint":false},{"pmid":"27943279","id":"PMC_27943279","title":"Biology and therapeutic implications of VEGF-A splice isoforms and single-nucleotide polymorphisms in colorectal cancer.","date":"2017","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/27943279","citation_count":36,"is_preprint":false},{"pmid":"30016789","id":"PMC_30016789","title":"VEGF-A and VEGF-B Coordinate the Arteriogenesis to Repair the Infarcted Heart with Vagus Nerve Stimulation.","date":"2018","source":"Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/30016789","citation_count":36,"is_preprint":false},{"pmid":"16565972","id":"PMC_16565972","title":"Differential expression of VEGF-A and angiopoietins in cartilage tumors and regulation by interleukin-1beta.","date":"2006","source":"Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/16565972","citation_count":35,"is_preprint":false},{"pmid":"25692621","id":"PMC_25692621","title":"Interplay between VEGF-A and cMET signaling in human vestibular schwannomas and schwann cells.","date":"2015","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/25692621","citation_count":34,"is_preprint":false},{"pmid":"20144626","id":"PMC_20144626","title":"Endothelial cell activation in a VEGF-A gradient: relevance to cell fate decisions.","date":"2010","source":"Microvascular research","url":"https://pubmed.ncbi.nlm.nih.gov/20144626","citation_count":34,"is_preprint":false},{"pmid":"36496136","id":"PMC_36496136","title":"RNA-binding motif 4 promotes angiogenesis in HCC by selectively activating VEGF-A expression.","date":"2022","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/36496136","citation_count":32,"is_preprint":false},{"pmid":"28899659","id":"PMC_28899659","title":"BRG1 promotes VEGF-A expression and angiogenesis in human colorectal cancer cells.","date":"2017","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/28899659","citation_count":32,"is_preprint":false},{"pmid":"25586350","id":"PMC_25586350","title":"Adiponectin inhibits VEGF-A in prostate cancer cells.","date":"2015","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/25586350","citation_count":32,"is_preprint":false},{"pmid":"38522817","id":"PMC_38522817","title":"Extracellular vesicles derived from dendritic cells loaded with VEGF-A siRNA and doxorubicin reduce glioma angiogenesis in vitro.","date":"2024","source":"Journal of controlled release : official journal of the Controlled Release Society","url":"https://pubmed.ncbi.nlm.nih.gov/38522817","citation_count":31,"is_preprint":false},{"pmid":"28400373","id":"PMC_28400373","title":"Association between VEGF-A and VEGFR-2 polymorphisms and response to treatment of neovascular AMD with anti-VEGF agents: a meta-analysis.","date":"2016","source":"The British journal of ophthalmology","url":"https://pubmed.ncbi.nlm.nih.gov/28400373","citation_count":31,"is_preprint":false},{"pmid":"38561075","id":"PMC_38561075","title":"Dual-layer conduit containing VEGF-A - Transfected Schwann cells promotes peripheral nerve regeneration via angiogenesis.","date":"2024","source":"Acta biomaterialia","url":"https://pubmed.ncbi.nlm.nih.gov/38561075","citation_count":30,"is_preprint":false},{"pmid":"29738889","id":"PMC_29738889","title":"Molecular dynamics-based model of VEGF-A and its heparin interactions.","date":"2018","source":"Journal of molecular graphics & modelling","url":"https://pubmed.ncbi.nlm.nih.gov/29738889","citation_count":29,"is_preprint":false},{"pmid":"33770315","id":"PMC_33770315","title":"CXCL1 stimulates decidual angiogenesis via the VEGF-A pathway during the first trimester of pregnancy.","date":"2021","source":"Molecular and cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/33770315","citation_count":29,"is_preprint":false},{"pmid":"34037749","id":"PMC_34037749","title":"Endothelial Insulin Receptors Promote VEGF-A Signaling via ERK1/2 and Sprouting Angiogenesis.","date":"2021","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/34037749","citation_count":28,"is_preprint":false},{"pmid":"19424629","id":"PMC_19424629","title":"Expression analysis of VEGF-A and VEGF-B: relationship with clinicopathological parameters in bladder cancer.","date":"2009","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/19424629","citation_count":28,"is_preprint":false},{"pmid":"33057033","id":"PMC_33057033","title":"Osteocyte Vegf-a contributes to myeloma-associated angiogenesis and is regulated by Fgf23.","date":"2020","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33057033","citation_count":27,"is_preprint":false},{"pmid":"24227910","id":"PMC_24227910","title":"Expression of VEGF-A, Otx homeobox and p53 family genes in proliferative vitreoretinopathy.","date":"2013","source":"Mediators of inflammation","url":"https://pubmed.ncbi.nlm.nih.gov/24227910","citation_count":27,"is_preprint":false},{"pmid":"27668980","id":"PMC_27668980","title":"VEGF-A and VEGFR1 SNPs associate with preeclampsia in a Philippine population.","date":"2016","source":"Clinical and experimental hypertension (New York, N.Y. : 1993)","url":"https://pubmed.ncbi.nlm.nih.gov/27668980","citation_count":27,"is_preprint":false},{"pmid":"28963126","id":"PMC_28963126","title":"EGF regulation of proximal tubule cell proliferation and VEGF-A secretion.","date":"2017","source":"Physiological reports","url":"https://pubmed.ncbi.nlm.nih.gov/28963126","citation_count":26,"is_preprint":false},{"pmid":"34823219","id":"PMC_34823219","title":"Substrate stiffening promotes VEGF-A functions via the PI3K/Akt/mTOR pathway.","date":"2021","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/34823219","citation_count":25,"is_preprint":false},{"pmid":"32300208","id":"PMC_32300208","title":"Cigarette smoking induces human CCR6+Th17 lymphocytes senescence and VEGF-A secretion.","date":"2020","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/32300208","citation_count":25,"is_preprint":false},{"pmid":"23081980","id":"PMC_23081980","title":"Expression and role of VEGF--a in the ciliary body.","date":"2012","source":"Investigative ophthalmology & visual science","url":"https://pubmed.ncbi.nlm.nih.gov/23081980","citation_count":25,"is_preprint":false},{"pmid":"26267146","id":"PMC_26267146","title":"Hypoxia-induced DNp73 stabilization regulates Vegf-A expression and tumor angiogenesis similar to TAp73.","date":"2015","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/26267146","citation_count":25,"is_preprint":false},{"pmid":"29789879","id":"PMC_29789879","title":"Vegf-A mRNA transfection as a novel approach to improve mouse and human islet graft revascularisation.","date":"2018","source":"Diabetologia","url":"https://pubmed.ncbi.nlm.nih.gov/29789879","citation_count":24,"is_preprint":false},{"pmid":"30821887","id":"PMC_30821887","title":"Local VEGF-A blockade modulates the microenvironment of the corneal graft bed.","date":"2019","source":"American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons","url":"https://pubmed.ncbi.nlm.nih.gov/30821887","citation_count":24,"is_preprint":false},{"pmid":"15453096","id":"PMC_15453096","title":"Role of Ang1 and its interaction with VEGF-A in astrocytomas.","date":"2004","source":"Journal of neuropathology and experimental neurology","url":"https://pubmed.ncbi.nlm.nih.gov/15453096","citation_count":24,"is_preprint":false},{"pmid":"36968223","id":"PMC_36968223","title":"A Phase I Trial of VEGF-A Inhibition Combined with PD-L1 Blockade for Recurrent Glioblastoma.","date":"2023","source":"Cancer research communications","url":"https://pubmed.ncbi.nlm.nih.gov/36968223","citation_count":23,"is_preprint":false},{"pmid":"26300505","id":"PMC_26300505","title":"Cellular and molecular aspects of diabetic nephropathy; the role of VEGF-A.","date":"2015","source":"Nefrologia : publicacion oficial de la Sociedad Espanola Nefrologia","url":"https://pubmed.ncbi.nlm.nih.gov/26300505","citation_count":22,"is_preprint":false},{"pmid":"15458440","id":"PMC_15458440","title":"Regulation of vascular permeability factor/vascular endothelial growth factor (VPF/VEGF-A) expression in podocytes.","date":"2004","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/15458440","citation_count":22,"is_preprint":false},{"pmid":"17203176","id":"PMC_17203176","title":"Distinct roles of VEGF-A and VEGF-C in tumour metastasis of gastric carcinoma.","date":"2007","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/17203176","citation_count":22,"is_preprint":false},{"pmid":"33401382","id":"PMC_33401382","title":"Diagnostic Value of VEGF-A, VEGFR-1 and VEGFR-2 in Feline Mammary Carcinoma.","date":"2021","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/33401382","citation_count":21,"is_preprint":false},{"pmid":"33865457","id":"PMC_33865457","title":"Anti-VEGF therapy prevents Müller intracellular edema by decreasing VEGF-A in diabetic retinopathy.","date":"2021","source":"Eye and vision (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/33865457","citation_count":21,"is_preprint":false},{"pmid":"25414781","id":"PMC_25414781","title":"New Insights into VEGF-A Alternative Splicing: Key Regulatory Switching in the Pathological Process.","date":"2014","source":"Avicenna journal of medical biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/25414781","citation_count":21,"is_preprint":false},{"pmid":"29462869","id":"PMC_29462869","title":"Modulation of VEGF-A Alternative Splicing as a Novel Treatment in Chronic Kidney Disease.","date":"2018","source":"Genes","url":"https://pubmed.ncbi.nlm.nih.gov/29462869","citation_count":20,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":54090,"output_tokens":7707,"usd":0.138937,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":17231,"output_tokens":5813,"usd":0.11574,"stage2_stop_reason":"end_turn"},"total_usd":0.254677,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1993,\n      \"finding\": \"VEGF-A (VPF/VEGF) acts directly on endothelial cells via two high-affinity tyrosine kinase receptors to activate phospholipase C and induce intracellular calcium transients; it is secreted as a 34–42 kDa heparin-binding, dimeric, disulfide-bonded glycoprotein.\",\n      \"method\": \"Receptor binding assays, functional in vitro assays (phospholipase C activation, calcium transients), biochemical characterization\",\n      \"journal\": \"Cancer metastasis reviews\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — direct in vitro biochemical assays with multiple orthogonal methods, replicated across multiple labs over time\",\n      \"pmids\": [\"8281615\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Akt signaling is both necessary and sufficient for VEGF-A-induced vascular permeability in vivo; dominant-negative Akt blocks VEGF-induced permeability, and constitutively active Akt mimics it; this Akt-mediated permeability requires eNOS activity.\",\n      \"method\": \"Adenovirus-mediated gene transfer of dominant-negative and constitutively active Akt in vivo (Miles assay), eNOS inhibitor (L-NAME) blockade\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vivo gain- and loss-of-function with mechanistic inhibitor validation, single lab but multiple orthogonal approaches\",\n      \"pmids\": [\"12459464\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"VEGF-A enhances endothelial PDGF-B expression; combined with FGF-2 (which enhances mural PDGFRβ expression), this directs endogenous PDGF-B–PDGFRβ signaling to recruit mural cells and form functional neovasculature; abrogation by anti-PDGFRβ antibody confirmed this mechanism.\",\n      \"method\": \"In vitro VEGFR2+ ESC-derived cell stimulation assays, Matrigel plug assay in vivo, neutralizing antibody blockade\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal in vitro and in vivo experiments with neutralizing antibody epistasis, single lab\",\n      \"pmids\": [\"16105884\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"VEGF-A stimulates chemotactic migration of human mesenchymal progenitor cells via VEGFR-1 (Flt-1); VEGFR-1 and VEGFR-2 are activated upon ligand stimulation in these cells, and the effect is mediated by VEGFR-1 since PlGF-1 (a VEGFR-1-selective ligand) but not VEGF-E or VEGF-C produced the same effect.\",\n      \"method\": \"In vitro kinase assay, Boyden chamber chemotaxis assay, quantitative RT-PCR for receptor expression\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — receptor activation confirmed by in vitro kinase assay, isoform selectivity used as epistatic evidence, single lab\",\n      \"pmids\": [\"16005848\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"VEGF-A overexpression in primary tumors induces lymphangiogenesis in sentinel lymph nodes even before metastasis occurs; VEGF-A acts on VEGFR-2-expressing lymphatic vessels to drive both tumor-associated and sentinel lymph node lymphangiogenesis.\",\n      \"method\": \"VEGF-A skin-specific transgenic mice, chemically induced carcinogenesis, immunohistochemistry for lymphatic markers\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — transgenic gain-of-function in vivo model with histological readout, well-controlled study\",\n      \"pmids\": [\"15809353\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"VEGF-A gene targeting in thymus epithelial cells (cortical and medullary TECs) disrupts the organ-typical thymus vascular architecture, causing hypovascularization, demonstrating that TEC-derived VEGF-A is required for normal thymus vascular morphogenesis.\",\n      \"method\": \"Conditional gene targeting via nude mouse blastocyst complementation, histological analysis of vascular architecture\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional knockout with defined cellular source and clear structural phenotype, rigorous genetic approach\",\n      \"pmids\": [\"16027358\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"VEGF165 protein is cleaved by plasmin at Arg110/Ala111 in the wound microenvironment, reducing its mitogenic activity; mutagenesis of this cleavage site preserves structural integrity and increases angiogenic potency in an impaired healing mouse model. Additionally, soluble VEGFR-1 (sVEGFR-1) acts as an endogenous inhibitor of VEGF-A in non-healing wounds.\",\n      \"method\": \"Protease cleavage assay, site-directed mutagenesis, in vivo impaired healing mouse model, ELISA for sVEGFR-1\",\n      \"journal\": \"The journal of investigative dermatology. Symposium proceedings\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro cleavage assay with mutagenesis validated in vivo, single lab\",\n      \"pmids\": [\"17069014\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"IFN-γ suppresses monocyte VEGF-A translation via the GAIT (IFN-γ-activated inhibitor of translation) complex, which binds a specific element in the VEGF-A 3′UTR; although IFN-γ induces VEGF-A mRNA, the GAIT complex delays and silences translation, reducing VEGF-A protein and angiogenic activity.\",\n      \"method\": \"mRNA-protein interaction studies (EMSA/RNA pulldown), translation reporter assays, angiogenesis functional assays\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — direct biochemical identification of GAIT complex binding to VEGF-A 3′UTR, functional translation silencing demonstrated with multiple orthogonal methods, single lab\",\n      \"pmids\": [\"17611605\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"HSV-1-induced corneal lymphangiogenesis is strictly dependent on VEGF-A/VEGFR-2 signaling (not VEGFR-3 ligands); infected epithelial cells (not macrophages) are the primary source of VEGF-A during infection, as identified using VEGF-A reporter transgenic mice.\",\n      \"method\": \"VEGF-A reporter transgenic mice, VEGFR-2 and VEGFR-3 blocking studies, macrophage depletion, in vivo corneal infection model\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple genetic and pharmacological epistasis approaches in vivo, reporter-based cellular source identification, single lab\",\n      \"pmids\": [\"20026662\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"VEGF-A and HGF cooperate in endothelial angiogenesis by synergistically activating ERK1/2 and p38 kinases downstream of their respective receptors (VEGFR-2 and c-Met), which do not physically associate or transphosphorylate each other; VEGF-A activates Rho-dependent cytoskeletal remodeling while HGF activates Rac-dependent remodeling.\",\n      \"method\": \"Co-immunoprecipitation (negative for receptor association), MAPK activation kinetics assays, Rho/Rac GTPase activity assays, in vitro tube formation\",\n      \"journal\": \"Biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple signaling pathway readouts with negative epistasis for direct receptor interaction, single lab\",\n      \"pmids\": [\"19281453\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Autocrine VEGF-A and VEGF-C in human podocytes both activate anti-apoptotic PI3K/AKT and suppress pro-apoptotic p38MAPK via VEGFR-2; ablation of VEGF-A or VEGF-C, or treatment with bevacizumab or VEGFR-2/-3 inhibitors, reduces podocyte survival. Exogenous VEGF-C can substitute for VEGF-A and vice versa in maintaining survival signaling.\",\n      \"method\": \"siRNA knockdown, bevacizumab treatment, VEGFR-2/-3 tyrosine kinase inhibition, Western blot for PI3K/AKT and p38MAPK activation, viability assays\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal siRNA knockdown and pharmacological inhibition with defined signaling readouts, single lab\",\n      \"pmids\": [\"19828679\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"VEGF-A expression in osteoclasts is regulated by HIF-1α, which itself is induced downstream of NF-κB activation by RANKL; NF-κB inhibition suppresses HIF-1α mRNA, and HIF-1α inhibition decreases VEGF-A mRNA, placing NF-κB upstream of HIF-1α upstream of VEGF-A in osteoclast signaling.\",\n      \"method\": \"Specific transcription factor inhibitors (NF-κB, AP-1, NFATc1, HIF-1), RT-PCR, conditioned media VEGF-A secretion assay\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — pharmacological inhibitor epistasis in cell culture with mRNA and protein readouts, single lab\",\n      \"pmids\": [\"20432243\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"IRE1α, PERK, and ATF6α of the unfolded protein response (UPR) each regulate VEGF-A mRNA transcription via their respective downstream transcription factors (spliced XBP-1, ATF4, and cleaved ATF6); loss of these UPR components in mouse embryonic fibroblasts or ATF6α-knockdown cells attenuates VEGF-A induction under ER stress.\",\n      \"method\": \"Genetic knockouts (Ire1α−/−, Perk−/− MEFs), siRNA knockdown (ATF6α), rescue experiments, qRT-PCR\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple genetic loss-of-function models with rescue experiments and transcriptional readout, identifies three parallel UPR pathways\",\n      \"pmids\": [\"20221394\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"VEGF-A-induced signaling through VEGFR-2 is concentration- and duration-dependent: PLCγ/ERK1/2 (p44/p42 MAPK) pathway transduces graded VEGF-A concentration information, whereas the parallel AKT pathway does not; longer exposure duration does not compensate for low VEGF-A concentration, indicating these parameters are not equivalent.\",\n      \"method\": \"Quantitative VEGFR2 autophosphorylation assays, signal kinase activation assays at graded VEGF-A concentrations, immediate-early gene induction analysis\",\n      \"journal\": \"Microvascular research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic dose-response and time-course signaling experiments in endothelial cells, single lab\",\n      \"pmids\": [\"20144626\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"VEGF-A induces degradation of versican in venular basement membranes during mother vessel formation by upregulating ADAMTS-1 (a versican-cleaving protease) and MMP-15 (which activates ADAMTS-1) in endothelial cells.\",\n      \"method\": \"Immunohistochemistry, Western blot, in vivo adenoviral VEGF-A(164) expression model, in vitro VEGF-A endothelial cell stimulation\",\n      \"journal\": \"The journal of histochemistry and cytochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro and in vivo corroborating experiments identifying protease induction mechanism, single lab\",\n      \"pmids\": [\"21411713\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"VEGF-A produced in the tumor microenvironment enhances expression of PD-1, Tim-3, and other inhibitory checkpoints on CD8+ T cells, promoting T cell exhaustion; this effect can be reverted by anti-angiogenic agents targeting VEGF-A/VEGFR.\",\n      \"method\": \"In vivo tumor models, flow cytometry for inhibitory receptor expression, anti-VEGF-A/VEGFR treatment reversal experiments\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo loss-of-function with defined cellular phenotypic readout (checkpoint receptor expression), single lab\",\n      \"pmids\": [\"25601652\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"TGF-β1 regulates peritoneal VEGF-A expression through an ID1-dependent pathway in mesothelial cells; siRNA knockdown of ID1 abolishes TGF-β1-induced VEGF-A upregulation, placing ID1 as an intermediary between TGF-β1 and VEGF-A transcription.\",\n      \"method\": \"siRNA knockdown of ID1, qRT-PCR and ELISA for VEGF-A, TGF-β1 stimulation of primary peritoneal and immortalized mesothelial cells\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA epistasis with protein-level readout, multiple cell types tested, single lab\",\n      \"pmids\": [\"26577912\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"VEGF-A165a sensitizes peripheral nociceptive neurons through VEGFR-2 and a TRPV1-dependent mechanism to enhance pain signaling; VEGF-A165b blocks this effect. After nerve injury, SRPK1-dependent pre-mRNA splicing shifts the isoform balance toward VEGF-Axxxa over VEGF-Axxxb, promoting neuropathic pain. Pharmacological inhibition of SRPK1 reverses this shift and alleviates pain.\",\n      \"method\": \"In vivo pain behavioral assays in rats and mice, VEGFR2 and TRPV1 pharmacological blockade, SRPK1 inhibition, exogenous VEGF-A165b treatment\",\n      \"journal\": \"Neurobiology of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo pharmacological epistasis identifying VEGFR2-TRPV1 pathway and SRPK1 splicing regulation, single lab\",\n      \"pmids\": [\"25151644\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Different VEGF-A isoforms (VEGF-A165, VEGF-A121, VEGF-A145) promote distinct patterns of VEGFR-2 endocytosis, differential ubiquitylation, and differential signal transduction; disruption of clathrin-dependent endocytosis blocks isoform-specific VEGFR-2 activation and causes depletion of membrane-bound VEGFR1 and VEGFR2.\",\n      \"method\": \"Clathrin endocytosis disruption, receptor internalization assays, Western blot for ubiquitylation and signaling, receptor proteolysis assays\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple isoforms compared with receptor trafficking and signaling readouts, clathrin inhibition epistasis, single lab\",\n      \"pmids\": [\"27044325\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"VEGF-A isoforms bind VEGFR2 with similar affinities but differ in interactions with co-receptor Neuropilin-1 and heparan sulfate proteoglycans (HSPGs); alternative splicing at exons 5–7 controls heparin-binding and bioavailability, while alternative 3′ splice-site selection at exon 8 generates VEGFxxxb isoforms with distinct downstream signaling.\",\n      \"method\": \"Receptor binding assays, downstream signaling analysis, VEGF-A isoform pharmacological comparison (review synthesizing experimental data)\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Strong — review synthesizing experimental results from multiple labs; individual mechanistic findings supported by cited experiments\",\n      \"pmids\": [\"29690653\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Molecular dynamics and circular dichroism modeling reveals that the VEGF-A heparin-binding domain (HBD) forms sandwich-like HBD-heparin-HBD structures that regulate the mutual disposition of HBDs and affect VEGF-A-mediated signaling; conformational flexibility of the 12-amino acid interdomain linker contributes to this regulation.\",\n      \"method\": \"Molecular docking, molecular dynamics simulation, circular dichroism spectroscopy\",\n      \"journal\": \"Journal of molecular graphics & modelling\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational modeling with only CD spectroscopy experimental validation, no functional mutagenesis\",\n      \"pmids\": [\"29738889\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TRF2 promotes VEGF-A secretion and angiogenesis by transcriptionally upregulating SULF2 (an endoglucosamine-6-sulfatase) via binding to a distal regulatory element; SULF2 then modifies heparan sulfate proteoglycans to release membrane-associated VEGF-A, increasing its availability.\",\n      \"method\": \"Luminex multiplexed secretome profiling, TRF2 overexpression/knockdown, ChIP for TRF2 binding, SULF2 knockdown epistasis, endothelial differentiation assay\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP identifies regulatory element, SULF2 epistasis confirms pathway, secretome profiling, single lab\",\n      \"pmids\": [\"30698737\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Granuloma macrophages produce VEGF-A that recruits immune cells to granulomas via a non-angiogenic pathway; selective VEGF-A blockade in myeloid cells or pharmaceutical inhibition reduces granulomatous inflammation and improves survival in Mycobacterium tuberculosis-infected mice without altering host protection.\",\n      \"method\": \"Conditional myeloid-specific VEGF-A knockout, granuloma transplantation, pharmaceutical VEGF-A inhibition, in vivo Mtb and BCG infection models\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional genetic knockout combined with pharmacological inhibition and transplantation epistasis in multiple infection models\",\n      \"pmids\": [\"31091450\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Endothelial insulin receptors are required for appropriate VEGF-A signal transduction from VEGFR-2 to ERK1/2; insulin receptor haploinsufficiency impairs VEGFR-2 internalization (required for ERK1/2 signaling) and abolishes VEGF-A-induced ERK1/2 activation, while VEGF-A signaling to Akt and eNOS remains intact.\",\n      \"method\": \"Whole-body and endothelium-restricted insulin receptor haploinsufficient mice, shRNA-mediated Insr knockdown in HUVECs, Western blot for ERK1/2 and Akt, VEGFR-2 internalization assays, in vitro and in vivo angiogenesis assays\",\n      \"journal\": \"Endocrinology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple genetic models (whole-body and endothelium-restricted KO) plus shRNA knockdown with defined signaling pathway dissection, two orthogonal experimental systems\",\n      \"pmids\": [\"34037749\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Laminin and α3β1 integrin signaling regulates constitutive VEGF-A expression in podocytes via a non-hypoxic HIF-α-dependent mechanism; classical PKC is a potential intermediary; HIF-α activity in podocytes is increased independently of hypoxia and drives VEGF-A promoter activity.\",\n      \"method\": \"VEGF-A promoter-luciferase reporter assay, quantitative RT-PCR, ELISA, immunoprecipitation for HIF-α/p300 association, extracellular matrix stimulation\",\n      \"journal\": \"Kidney international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reporter assay with matrix-specific conditions and HIF-α co-immunoprecipitation, single lab preliminary study\",\n      \"pmids\": [\"15458440\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"VEGF-A-regulated gene transcription in endothelial cells is controlled at two levels: RNA polymerase II pausing at promoters and productive elongation; approximately half of VEGF-A-regulated promoters show paused Pol II, and transition to productive elongation is a major activation mechanism for virtually all VEGF-regulated genes.\",\n      \"method\": \"Genome-wide GRO-Seq (global run-on sequencing), tethered conformation capture (TCC) chromatin interaction mapping in primary HAECs and HUVECs\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — genome-wide mechanistic transcriptional assay (GRO-Seq) with chromatin interaction mapping, two primary cell types, single lab\",\n      \"pmids\": [\"25352550\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"WISP-1 promotes VEGF-A expression in osteosarcoma cells and angiogenesis through activation of the FAK/JNK/HIF-1α signaling pathway and down-regulation of miR-381; inhibitors of FAK, JNK, or HIF-1α, or HIF-1α siRNA, abolish WISP-1-induced VEGF-A expression.\",\n      \"method\": \"Pathway inhibitors (FAK, JNK, HIF-1α), siRNA knockdown, in vitro EPC migration/tube formation, in vivo tumor xenograft, immunohistochemistry\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological and genetic epistasis with in vitro and in vivo validation, single lab\",\n      \"pmids\": [\"28406476\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"VEGF-A can signal intracellularly (intracrine signaling) within cells expressing both VEGF-A and its receptors, regulating cell growth, survival, and metabolism independently of secretion; this intracrine mode involves VEGFR1 and VEGFR2 expressed intracellularly.\",\n      \"method\": \"Review synthesizing loss-of-function and localization experiments from multiple studies demonstrating intracellular VEGF-A/receptor complexes\",\n      \"journal\": \"Biomolecules\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — review synthesizing others' findings; individual mechanistic experiments not described in the abstract itself\",\n      \"pmids\": [\"33478167\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"VEGF-A expressed by pigmented ciliary epithelium maintains ciliary body homeostasis; systemic VEGF-A neutralization (via adenoviral sFlt1 overexpression) causes thinning of nonpigmented epithelium, vacuolization of pigmented epithelium, loss of capillary fenestrations, thrombosis, and decreased intraocular pressure, indicating a required role in ciliary body vascular and epithelial maintenance.\",\n      \"method\": \"VEGF-A neutralization via adenoviral sFlt1, VEGF-LacZ reporter mice, VEGFR2 localization by immunostaining, light microscopy and TEM, intraocular pressure measurement\",\n      \"journal\": \"Investigative ophthalmology & visual science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo VEGF-A neutralization with defined structural and functional phenotypic readouts, reporter-based localization, single lab\",\n      \"pmids\": [\"23081980\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In multiple myeloma, FGF23 produced by osteocytes in response to direct contact with myeloma cells upregulates VEGF-A expression in osteocytes; VEGF-A knockdown in osteocytes or VEGF-A neutralization in conditioned media completely abolishes the increased endothelial tube formation induced by myeloma–osteocyte co-culture.\",\n      \"method\": \"siRNA knockdown of Vegf-a in osteoclasts/primary osteocytes, VEGF-A neutralization, co-culture endothelial tube formation assay, Fgf23 deletion, in vivo myeloma mouse model with vessel area quantification\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockdown and neutralization epistasis in vitro and in vivo with mechanistic Fgf23–Vegf-a pathway link, single lab\",\n      \"pmids\": [\"33057033\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"VEGF-A is a secreted dimeric glycoprotein that acts primarily through two high-affinity tyrosine kinase receptors (VEGFR-1 and VEGFR-2) to activate endothelial cells via PLCγ/ERK1/2, PI3K/Akt/eNOS, and other downstream pathways, driving angiogenesis, vascular permeability, lymphangiogenesis, and immune cell modulation; multiple alternatively spliced isoforms differ in heparin-binding, bioavailability, Neuropilin-1 co-receptor interaction, and VEGFR-2 trafficking/signaling outputs; VEGF-A translation is post-transcriptionally silenced by the IFN-γ–induced GAIT complex binding to its 3′UTR; its transcription is regulated by HIF-1α (under hypoxia and non-hypoxically via integrin signaling in podocytes), UPR sensors (IRE1α-XBP-1, PERK-ATF4, ATF6α), NF-κB, and chromatin-level Pol II pausing/elongation control; extracellular VEGF-A bioavailability is further regulated by plasmin cleavage, SULF2-mediated heparan sulfate modification, and soluble VEGFR-1 sequestration; VEGF-A also drives non-angiogenic functions including T-cell exhaustion via PD-1/Tim-3 upregulation, immune-cell recruitment to granulomas, nociceptor sensitization via VEGFR-2/TRPV1, and autocrine podocyte survival via PI3K/AKT/p38MAPK.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"VEGF-A is a secreted, disulfide-bonded dimeric heparin-binding glycoprotein that signals through two high-affinity endothelial tyrosine kinase receptors (VEGFR-1/Flt-1 and VEGFR-2) to drive angiogenesis, vascular permeability, lymphangiogenesis, and tissue-specific vascular maintenance [#0, #5, #28]. Receptor engagement activates parallel downstream cascades whose outputs are non-equivalent: VEGFR-2-coupled PLC\\u03b3/ERK1/2 signaling transduces graded VEGF-A concentration information, whereas the Akt arm operates as a concentration-independent switch that is both necessary and sufficient for vascular permeability through eNOS [#1, #13]. Efficient ERK1/2 activation requires VEGFR-2 internalization, which itself depends on the endothelial insulin receptor, while signaling to Akt/eNOS proceeds independently [#23]. Beyond endothelium, VEGF-A drives mural-cell recruitment by inducing endothelial PDGF-B [#2], promotes chemotaxis of mesenchymal progenitors via VEGFR-1 [#3], remodels the venular basement membrane by upregulating ADAMTS-1 and MMP-15 [#14], and supports autocrine podocyte survival via VEGFR-2-driven PI3K/AKT with suppression of p38MAPK [#10]. VEGF-A also executes non-angiogenic programs: it sensitizes nociceptors through a VEGFR-2/TRPV1 mechanism [#17], promotes CD8+ T-cell exhaustion by upregulating PD-1 and Tim-3 [#15], and recruits immune cells to tuberculous granulomas from a myeloid source [#22]. Its output is shaped extensively by alternative splicing\\u2014exon 5\\u20137 selection tunes heparin/HSPG and Neuropilin-1 binding and bioavailability, while exon 8 3\\u2032-splice-site choice generates VEGFxxxb isoforms\\u2014and distinct isoforms drive distinct VEGFR-2 endocytosis, ubiquitylation, and signaling [#18, #19, #17, #18]. Transcription is governed by HIF-1\\u03b1 (under hypoxia and non-hypoxically via integrin signaling), NF-\\u03baB-to-HIF-1\\u03b1 cascades, the UPR sensors IRE1\\u03b1-XBP-1/PERK-ATF4/ATF6\\u03b1, and Pol II pausing-to-elongation control [#11, #12, #24, #25], while translation is silenced post-transcriptionally by the IFN-\\u03b3-induced GAIT complex binding the 3\\u2032UTR [#7]. Extracellular bioavailability is further set by plasmin cleavage at Arg110/Ala111, soluble VEGFR-1 sequestration, and SULF2-mediated heparan sulfate modification that releases matrix-bound ligand [#6, #21].\",\n  \"teleology\": [\n    {\n      \"year\": 1993,\n      \"claim\": \"Established VEGF-A as a secreted dimeric heparin-binding glycoprotein that acts directly on endothelial cells through two high-affinity tyrosine kinase receptors, defining the basic receptor-ligand axis.\",\n      \"evidence\": \"Receptor binding and functional in vitro assays (PLC activation, calcium transients), biochemical characterization\",\n      \"pmids\": [\"8281615\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve which receptor mediates which downstream output\", \"No isoform-specific signaling distinguished\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Showed that the Akt arm is both necessary and sufficient for VEGF-A-induced vascular permeability and acts through eNOS, separating permeability control from other VEGF outputs.\",\n      \"evidence\": \"Adenoviral dominant-negative/constitutively active Akt in vivo Miles assay with eNOS inhibitor blockade\",\n      \"pmids\": [\"12459464\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address how Akt is selectively activated downstream of which receptor\", \"Relationship to ERK1/2 arm not defined\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Demonstrated non-hypoxic transcriptional control of VEGF-A by laminin/\\u03b13\\u03b21 integrin signaling acting through HIF-\\u03b1 in podocytes, expanding HIF regulation beyond oxygen sensing.\",\n      \"evidence\": \"VEGF-A promoter-luciferase reporter, HIF-\\u03b1/p300 co-IP, matrix stimulation in podocytes\",\n      \"pmids\": [\"15458440\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"PKC intermediary only a candidate\", \"Mechanism of non-hypoxic HIF-\\u03b1 stabilization not resolved\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Defined cell-type-specific developmental and pathological angiogenic roles for VEGF-A: TEC-derived VEGF-A for thymus vascular morphogenesis, mural-cell recruitment via induced PDGF-B, VEGFR-1-mediated progenitor chemotaxis, and tumor-driven sentinel-node lymphangiogenesis.\",\n      \"evidence\": \"Conditional gene targeting, Matrigel plug and neutralizing antibody epistasis, Boyden chamber chemotaxis, VEGF-A transgenic mice with lymphatic IHC\",\n      \"pmids\": [\"16027358\", \"16105884\", \"16005848\", \"15809353\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor selectivity for lymphangiogenesis vs angiogenesis only partly resolved\", \"Crosstalk between PDGF-B and direct VEGF effects not fully separated\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Identified extracellular regulation of VEGF-A activity by plasmin cleavage at Arg110/Ala111 and soluble VEGFR-1 sequestration, establishing post-secretion control of angiogenic potency.\",\n      \"evidence\": \"Protease cleavage assay, site-directed mutagenesis, impaired-healing mouse model, sVEGFR-1 ELISA\",\n      \"pmids\": [\"17069014\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo contribution of cleavage vs sVEGFR-1 not quantitatively partitioned\", \"Other proteases not excluded\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Revealed post-transcriptional silencing of VEGF-A translation by the IFN-\\u03b3-induced GAIT complex binding its 3\\u2032UTR, decoupling mRNA induction from protein output.\",\n      \"evidence\": \"RNA-protein interaction (EMSA/pulldown), translation reporter and angiogenesis functional assays\",\n      \"pmids\": [\"17611605\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"GAIT regulation shown in monocytes; generality across cell types not established\", \"Kinetics of silencing onset not fully defined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Mapped non-angiogenic and cooperative signaling roles: autocrine podocyte survival via VEGFR-2/PI3K-AKT/p38, VEGF-A/HGF synergy via independent receptors with distinct Rho vs Rac remodeling, and VEGF-A/VEGFR-2 as the obligate driver of infection-induced corneal lymphangiogenesis.\",\n      \"evidence\": \"siRNA and pharmacological inhibition with signaling readouts, co-IP (negative for receptor association), Rho/Rac assays, VEGF-A reporter mice and receptor blockade in corneal infection\",\n      \"pmids\": [\"19828679\", \"19281453\", \"20026662\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"VEGF-A vs VEGF-C redundancy in podocytes not fully separated\", \"Synergy mechanism limited to MAPK readouts\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Resolved upstream transcriptional logic (NF-\\u03baB\\u2192HIF-1\\u03b1, three parallel UPR arms) and demonstrated that VEGFR-2 PLC\\u03b3/ERK1/2 encodes graded ligand concentration whereas Akt does not.\",\n      \"evidence\": \"Transcription factor inhibitor epistasis, Ire1\\u03b1/Perk knockout MEFs and ATF6\\u03b1 knockdown with rescue, quantitative dose-response/time-course VEGFR-2 signaling assays\",\n      \"pmids\": [\"20432243\", \"20221394\", \"20144626\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How concentration is decoded into distinct gene programs unresolved\", \"Integration of multiple UPR arms not quantified\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Showed VEGF-A drives basement-membrane remodeling during mother vessel formation by inducing endothelial ADAMTS-1 and its activator MMP-15 to degrade versican.\",\n      \"evidence\": \"IHC, Western blot, in vivo adenoviral VEGF-A164 and in vitro endothelial stimulation\",\n      \"pmids\": [\"21411713\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct functional requirement of versican cleavage for vessel formation not tested\", \"Single lab\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Established that VEGF-A-induced endothelial transcription is controlled by RNA Pol II pausing and transition to productive elongation across most regulated genes.\",\n      \"evidence\": \"Genome-wide GRO-Seq and tethered conformation capture in primary HAECs/HUVECs\",\n      \"pmids\": [\"25352550\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Pause-release factors recruited by VEGF signaling not identified\", \"Link to specific upstream kinase arms unmapped\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Extended VEGF-A function to immune suppression (CD8+ T-cell exhaustion via PD-1/Tim-3), nociceptor sensitization (VEGFR-2/TRPV1 with SRPK1-controlled isoform balance), and added ID1 as a TGF-\\u03b21\\u2192VEGF-A transcriptional intermediary.\",\n      \"evidence\": \"In vivo tumor models with checkpoint flow cytometry, pain behavioral assays with VEGFR2/TRPV1/SRPK1 manipulation, ID1 siRNA epistasis\",\n      \"pmids\": [\"25601652\", \"25151644\", \"26577912\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect VEGF-A action on T cells not fully resolved\", \"Tissue specificity of ID1 pathway unknown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrated isoform-specific control of VEGFR-2 endocytosis, ubiquitylation, and signaling, with clathrin-dependent internalization required for isoform-selective receptor activation.\",\n      \"evidence\": \"Clathrin disruption, receptor internalization and ubiquitylation assays, isoform comparison\",\n      \"pmids\": [\"27044325\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of differential trafficking not defined\", \"Downstream transcriptional consequences not mapped\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Consolidated the splicing-to-function logic: exon 5\\u20137 selection tunes heparin/HSPG and Neuropilin-1 binding and bioavailability, exon 8 generates VEGFxxxb isoforms, and HBD-heparin-HBD architecture modulates signaling.\",\n      \"evidence\": \"Review of receptor binding and signaling data; molecular dynamics and circular dichroism of the heparin-binding domain\",\n      \"pmids\": [\"29690653\", \"29738889\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"HBD conformational model rests on computation with limited experimental validation\", \"Functional mutagenesis of HBD disposition lacking\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified upstream bioavailability and inflammatory controls: TRF2-driven SULF2 transcription releases matrix-bound VEGF-A, and myeloid-derived VEGF-A recruits immune cells to tuberculous granulomas non-angiogenically.\",\n      \"evidence\": \"ChIP and SULF2 epistasis with secretome profiling; myeloid-specific VEGF-A knockout, granuloma transplantation, and pharmacological inhibition in Mtb/BCG infection\",\n      \"pmids\": [\"30698737\", \"31091450\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether HS remodeling generalizes beyond endothelial differentiation untested\", \"Immune-cell types recruited and receptor used in granulomas not fully defined\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined a niche signaling circuit in which myeloma-induced osteocyte FGF23 upregulates osteocyte VEGF-A to drive endothelial tube formation.\",\n      \"evidence\": \"Vegf-a siRNA and neutralization, co-culture tube formation, Fgf23 deletion, in vivo myeloma model\",\n      \"pmids\": [\"33057033\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor mediating FGF23\\u2192VEGF-A in osteocytes not identified\", \"Generality beyond myeloma niche unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated that endothelial insulin receptor is required for VEGFR-2 internalization and ERK1/2 signaling selectively, while Akt/eNOS signaling remains intact, linking metabolic receptor status to angiogenic signal routing; intracrine VEGF-A signaling was also proposed.\",\n      \"evidence\": \"Whole-body and endothelium-restricted Insr haploinsufficient mice plus HUVEC shRNA with signaling and internalization readouts; review synthesis for intracrine mode\",\n      \"pmids\": [\"34037749\", \"33478167\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism linking insulin receptor to VEGFR-2 endocytosis unresolved\", \"Intracrine model rests on review-level synthesis without primary mechanistic experiments here\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the multiple parallel transcriptional inputs, splicing decisions, receptor trafficking states, and extracellular bioavailability controls are integrated to produce a specific VEGF-A signaling output in a given cell remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model linking isoform identity to trafficking to transcriptional program\", \"Quantitative contribution of each bioavailability control in vivo unknown\", \"Intracrine signaling not mechanistically established in primary data\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [0, 3, 13, 18]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 1, 13]},\n      {\"term_id\": \"GO:0008201\", \"supporting_discovery_ids\": [0, 19, 20]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0, 6, 21]},\n      {\"term_id\": \"GO:0031012\", \"supporting_discovery_ids\": [14, 21]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 13, 23]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 4, 5, 14]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [15, 22]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [11, 12, 24, 25]},\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [7, 17]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"FLT1\", \"KDR\", \"NRP1\", \"FLT4\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}