{"gene":"VEGFC","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":1997,"finding":"Stepwise proteolytic processing of the VEGF-C precursor generates forms with progressively increased activity toward VEGFR-3; only fully processed mature VEGF-C can activate VEGFR-2. Mature VEGF-C binds VEGFR-3 (Kd ~135 pM) and VEGFR-2 (Kd ~410 pM), activates both receptors, increases vascular permeability, and stimulates endothelial cell migration and proliferation. Unlike other PDGF/VEGF family members, mature VEGF-C forms predominantly non-covalent homodimers.","method":"Recombinant protein production in yeast, receptor binding assays, receptor autophosphorylation assays, vascular permeability assays, endothelial cell migration and proliferation assays, biochemical characterization of processing intermediates","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with recombinant proteins, binding kinetics, functional receptor activation assays, multiple orthogonal methods in a single rigorous study","pmids":["9233800"],"is_preprint":false},{"year":1996,"finding":"Mouse VEGF-C is secreted as VEGFR-3 (Flt4)-binding polypeptides of 30–32 kDa and 22–23 kDa that preferentially stimulate VEGFR-3 autophosphorylation over VEGFR-2. In situ hybridization shows VEGF-C mRNA in mesenchymal cells near regions where lymphatic vessels sprout from embryonic veins, consistent with a paracrine role in lymphatic vessel development.","method":"Recombinant protein production, receptor autophosphorylation assay (VEGFR-3 vs VEGFR-2), in situ hybridization in mouse embryos","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — receptor activation assays plus spatial expression mapping, replicated across multiple labs","pmids":["9012504"],"is_preprint":false},{"year":1997,"finding":"VEGF-C is the first identified lymphangiogenic growth factor; it induces proliferation of lymphatic endothelial cells and formation of new lymphatic sinuses in the avian chorioallantoic membrane (CAM). VEGF, by contrast, is angiogenic but not lymphangiogenic in this model, and PlGF has neither activity.","method":"In vivo CAM assay with microinjection, immunohistochemistry, in situ hybridization for VEGFR-2 and VEGFR-3","journal":"Developmental biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct in vivo lymphangiogenesis assay with comparative growth factor controls, independently replicated","pmids":["9245515"],"is_preprint":false},{"year":2003,"finding":"The serine protease plasmin cleaves both NH2- and COOH-terminal propeptides from the VEGF homology domain of VEGF-C (and VEGF-D), generating a mature form with greatly enhanced binding and cross-linking of VEGFR-2 and VEGFR-3 compared to full-length material. Plasmin thereby activates VEGF-C.","method":"In vitro protease cleavage assay, receptor binding and cross-linking assays with full-length versus plasmin-processed VEGF-C","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct in vitro enzymatic reconstitution with biochemical characterization of cleavage products and receptor binding, independently replicated concept","pmids":["12963694"],"is_preprint":false},{"year":2006,"finding":"VEGF-C and VEGF-D directly interact with neuropilin-2 (NP2): VEGF-C binds NP2 in a heparin-independent manner while VEGF-D binding is heparin-dependent. The domains of VEGF-C and NP2 required for this interaction were mapped. NP2 co-internalizes with VEGFR-3 into endocytic vesicles of lymphatic endothelial cells upon VEGF-C or VEGF-D stimulation, and NP2 co-precipitates with VEGFR-3, indicating that NP2 participates in an active signaling complex with VEGFR-3.","method":"In vitro binding studies, domain mapping, co-immunoprecipitation, co-internalization imaging in lymphatic endothelial cells","journal":"FASEB journal","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal binding studies, co-IP, and co-internalization imaging; multiple orthogonal methods in single study","pmids":["16816121"],"is_preprint":false},{"year":2015,"finding":"Proteolytic maturation of VEGF-C releases a cryptic Nrp2-binding motif at the C-terminus, allowing specific engagement with the b1 domain of Nrp2. Crystal structure of the VEGF-C C-terminus in complex with Nrp2 ligand-binding domains was determined. The endogenous secreted splice form s9Nrp2 forms a stable dimer and potently inhibits VEGF-C/Nrp2 binding and downstream cellular signaling.","method":"X-ray crystallography, in vitro binding assays, cellular signaling assays","journal":"Structure (London, England : 1993)","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure with functional binding validation and cellular assays in a single rigorous study","pmids":["25752543"],"is_preprint":false},{"year":1998,"finding":"VEGF-C protein is stored in platelet alpha-granules and is released from platelets upon activation, together with beta-thromboglobulin. VEGF-C mRNA in peripheral blood is restricted to platelets and T cells.","method":"VEGF-C mRNA detection in blood cell fractions, VEGF-C protein release assay from activated platelets, co-release with beta-thromboglobulin as alpha-granule marker","journal":"Thrombosis and haemostasis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell fractionation, protein release assay, co-release with granule marker; single lab, two orthogonal methods","pmids":["9684805"],"is_preprint":false},{"year":2019,"finding":"VEGF-C is cleaved and activated by thrombin and plasmin generated during hemostasis. Platelets accelerate lymphatic growth after injury in vivo, and this platelet-enhanced lymphangiogenesis depends on platelet-derived VEGF-C (but not VEGF-D). VEGF-C, but not VEGF-D, is also the dominant factor for lymphangiogenesis after full-thickness skin excision.","method":"In vitro protease cleavage assays, tail-wounding mouse model, genetic studies with platelet-specific VEGF-C deletion, full-thickness excision wound model","journal":"Blood","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro biochemical reconstitution plus in vivo genetic studies with two wound models and multiple orthogonal approaches","pmids":["31562136"],"is_preprint":false},{"year":2015,"finding":"VEGF-C is required for maintenance of intestinal lymphatic vessels (lacteals) in adult mice. VEGF-C is expressed by smooth muscle cells adjacent to lacteals. Deletion of Vegfc in adult mice causes gradual atrophy of intestinal lymphatics, leading to defective lipid absorption, increased fecal excretion of dietary cholesterol and fatty acids, and resistance to diet-induced obesity.","method":"Conditional Vegfc gene deletion in adult mice, immunohistochemistry for lymphatic markers, lipid absorption assays, dietary challenge experiments","journal":"EMBO molecular medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional knockout with multiple phenotypic readouts and cell-of-origin identification; rigorous genetic study","pmids":["26459520"],"is_preprint":false},{"year":2000,"finding":"VEGF-C signaling via VEGFR-2 works synergistically with VEGF-A to enhance vascular bed formation. VEGF-C binding to VEGFR-3 sequesters VEGF-C away from VEGFR-2, thereby regulating VEGFR-2 signaling. In VEGFR-3-deficient embryos, excess VEGF-C signals through VEGFR-2, causing disturbed vasculogenesis and suppressed hematopoiesis.","method":"Para-aortic splanchnopleural mesoderm (P-Sp) coculture with OP9 stromal cells, soluble receptor competitive inhibition, VEGFR-3-deficient embryo explants","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and pharmacological epistasis in an ex vivo embryo coculture system; single lab with multiple experimental conditions","pmids":["11090062"],"is_preprint":false},{"year":2012,"finding":"FGF-2-induced lymphangiogenesis requires VEGFR-3 signaling; a VEGFR-3-neutralizing antibody markedly inhibits FGF-2-induced lymphangiogenesis. VEGFR-3-mediated signaling is a prerequisite for lymphatic tip cell formation in both FGF-2- and VEGF-C-induced lymphangiogenesis. FGFR-1 expressed in lymphatic endothelial cells is the receptor mediating FGF-2-induced lymphangiogenesis.","method":"Mouse corneal lymphangiogenesis assay, VEGFR-3 neutralizing antibody epistasis, tumor co-implantation model, in vitro lymphatic endothelial cell assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo epistasis with neutralizing antibody, multiple tumor and corneal models; single lab","pmids":["22967508"],"is_preprint":false},{"year":2014,"finding":"Hypoxia reduces VEGF-C cap-dependent translation via upregulation of hypophosphorylated 4E-BP1, but induces VEGF-C translation through an internal ribosome entry site (IRES)-dependent mechanism that is independent of HIF-1α. This IRES-dependent VEGF-C translation is higher in lymph node metastases than in primary tumors.","method":"IRES reporter assays, 4E-BP1 manipulation, HIF-1α knockdown, comparison of primary tumor vs. lymph node metastasis samples","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic dissection using reporter assays and genetic knockdown; multiple orthogonal approaches in single lab","pmids":["24388748"],"is_preprint":false},{"year":2009,"finding":"VEGF-C regulates capillary stabilization by controlling PDGF-B expression via VEGFR-3 in a paracrine mode during neovascularization. Blockade of VEGFR-3 inhibited FGF-2-mediated limb salvage and caused capillary dilation with mural cell dissociation, and VEGF-C and PDGF-B were mutually dependent (blocking VEGFR-3 decreased PDGF-B expression, and blocking PDGF-BB decreased VEGF-C expression).","method":"Murine ischemic hindlimb model, VEGFR-3 neutralizing antibody AFL-4, PDGF-BB blocking antibody, immunohistochemistry","journal":"American journal of physiology. Heart and circulatory physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo epistasis with neutralizing antibodies and multiple phenotypic readouts; single lab","pmids":["19734356"],"is_preprint":false},{"year":2016,"finding":"VEGF-C is required for the transition to fetal liver erythropoiesis. Embryonic Vegfc deletion causes defective fetal erythropoiesis with anemia and absence of enucleated red blood cells, reduced macrophages and erythroid cells in fetal liver due to decreased proliferation and increased apoptosis, decreased α4-integrin on erythro-myeloid progenitors (EMPs), and impaired EMP colonization of the fetal liver. Vegfc deletion from E10.5 onward or in adults does not affect definitive hematopoiesis.","method":"Conditional Vegfc deletion at E7.5, flow cytometry, immunohistochemistry, blood cell analysis, integrin expression assays","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional genetic knockout with temporally controlled deletion, multiple cellular and molecular readouts, defines a novel non-lymphangiogenic function","pmids":["27343251"],"is_preprint":false},{"year":2020,"finding":"VEGF-C maintains the integrity of the bone marrow perivascular niche. Global and conditional deletion of Vegfc from endothelial or LepR+ cells disrupts the BM perivascular niche and delays hematopoietic recovery after irradiation by decreasing endothelial proliferation and LepR+ cell regeneration. Exogenous AAV-delivered VEGF-C improves hematopoietic recovery by accelerating endothelial and LepR+ cell regeneration and increasing expression of hematopoietic regenerative factors.","method":"Conditional Vegfc gene deletion from endothelial and LepR+ cells, irradiation/transplantation model, AAV-mediated VEGF-C delivery, immunohistochemistry, flow cytometry","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional knockout with cell-type specificity, rescue experiment with exogenous VEGF-C, multiple orthogonal readouts","pmids":["32842144"],"is_preprint":false},{"year":2018,"finding":"VEGF-C acts as both a paracrine and autocrine pro-survival cytokine in glioblastoma, activating VEGFR-2 in an autocrine manner to promote tumor cell survival and sustain VEGFR2 activation that is bevacizumab-resistant. Targeting VEGF-C expression in vivo impairs tumor growth more effectively than bevacizumab treatment.","method":"RNA interference, exogenous ligand treatment, proximity ligation assay for VEGF-C/VEGFR2 interaction in tumor specimens, patient-derived xenograft in vivo experiments, matched pre/post-bevacizumab treatment cohort analysis","journal":"Neuro-oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNAi knockdown with functional readouts, proximity ligation assay for direct interaction, in vivo xenograft; single lab","pmids":["29939339"],"is_preprint":false},{"year":2020,"finding":"VEGF-C secreted by breast cancer cells that have undergone EMT promotes paracrine non-canonical GLI signaling activation in neighboring epithelial breast cancer cells via NRP2. Inhibiting VEGF-C in EMT cells or knocking down NRP2 in epithelial cancer cells disrupts the aggressive phenotypes (proliferation, migration, invasion, metastasis) imparted by EMT cells.","method":"VEGF-C knockdown in EMT cells, NRP2 knockdown in epithelial cells, co-culture systems, in vivo metastasis models, TCGA/GEO dataset correlation","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined pathway placement (VEGF-C/NRP2/GLI axis), in vivo validation; single lab","pmids":["33299122"],"is_preprint":false},{"year":2011,"finding":"C/EBP-δ transcription factor regulates VEGF-C and VEGFR-3 expression specifically in lymphatic endothelial cells (LECs). Genetic deletion of C/EBP-δ in mice dramatically reduces VEGF-C and VEGFR-3 in LECs, reduces lymphangiogenesis and pulmonary metastases. Forced VEGF-C expression (but not recombinant VEGF-C protein) rescues C/EBP-δ knockdown-induced LEC apoptosis, demonstrating autocrine VEGF-C signaling is essential for LEC survival. C/EBP-δ-induced VEGF-C expression requires HIF-1α.","method":"C/EBP-δ knockout mice, forced expression/knockdown in cultured LECs, HIF-1α blocking, rescue experiments with VEGF-C plasmid vs. recombinant protein","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockout in mice, forced expression/knockdown, mechanistic dissection with HIF-1α; single lab, multiple methods","pmids":["21666710"],"is_preprint":false},{"year":2009,"finding":"Autocrine VEGF-C (along with VEGF-A) in human podocytes is important for podocyte survival via VEGFR-2-mediated activation of PI3K/AKT (anti-apoptotic) and suppression of p38MAPK (pro-apoptotic). Exogenous VEGF-C can reverse the effect of VEGF-A neutralization, indicating functional overlap and complementarity between autocrine VEGF-A and VEGF-C in podocytes.","method":"siRNA knockdown of VEGF-A and VEGF-C in human podocytes, bevacizumab treatment, VEGFR-2/-3 tyrosine kinase inhibitor, phospho-AKT and p38MAPK western blotting, apoptosis assays","journal":"American journal of physiology. Renal physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA loss-of-function with defined signaling readouts and rescue experiments; single lab","pmids":["19828679"],"is_preprint":false},{"year":2006,"finding":"VEGF-C acts as an autocrine survival factor in cultured human podocytes via VEGFR-1 (not VEGFR-3, as VEGF-C did not autophosphorylate VEGFR-3 in these cells despite doing so in HMVECs). VEGF-C reduces intracellular calcium and cytotoxicity, reduces MAPK phosphorylation, and has no effect on Akt phosphorylation in podocytes.","method":"VEGF-C treatment of conditionally immortalized human podocytes, VEGFR-3 kinase inhibitor (MAZ51), SU-5416 (VEGFR-1-specific concentrations), immunoprecipitation for VEGFR-3 autophosphorylation, calcium imaging, cytotoxicity assays","journal":"American journal of physiology. Renal physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological inhibition with receptor-specific reagents and receptor immunoprecipitation; single lab, multiple assays","pmids":["16525158"],"is_preprint":false},{"year":2018,"finding":"VEGF-C drives bone lymphatic vessel formation via VEGFR-3 (but not VEGFR-2) signaling, as inhibition of VEGFR-3 but not VEGFR-2 prevented formation of bone lymphatics in Osx-tTA;TetO-Vegfc mice. VEGF-C overexpression in bone also promotes bone loss via increased osteoclast numbers, which is attenuated by an osteoclast inhibitor.","method":"Double-transgenic mouse model (Osx-tTA;TetO-Vegfc), VEGFR-selective antibody blockade, radiological and histological bone analysis, osteoclast inhibitor treatment","journal":"eLife","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic gain-of-function with receptor-selective inhibition epistasis; single lab","pmids":["29620526"],"is_preprint":false},{"year":2021,"finding":"OTUD3 deubiquitinase stabilizes ZFP36 by inhibiting FBXW7-mediated K48-linked polyubiquitination of ZFP36. ZFP36 binds the VEGF-C 3'-UTR and recruits the RNA-degrading complex to induce rapid VEGF-C mRNA decay. Nicotine downregulates OTUD3, leading to ZFP36 destabilization and increased VEGF-C production and lymphatic metastasis.","method":"Co-immunoprecipitation of OTUD3-ZFP36 interaction, ubiquitination assays, RNA decay assays, ZFP36-VEGF-C 3'-UTR binding assays, OTUD3 knockdown/overexpression in cancer cells, in vivo metastasis model","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — biochemical reconstitution of the ubiquitination pathway, RNA decay assays, direct 3'-UTR binding, in vivo validation; multiple orthogonal methods","pmids":["34853315"],"is_preprint":false},{"year":2020,"finding":"Functional VEGF-C is transported by extracellular vesicles (EVs) from endometriotic cells to lymphatic endothelial cells, enhancing their lymphangiogenic ability. VEGF-C expression in endometriotic cells is negatively regulated by COUP-TFII, and proinflammatory cytokines increase VEGF-C by suppressing COUP-TFII levels.","method":"EV isolation and functional transfer assay, COUP-TFII knockdown/overexpression, cytokine treatment, autotransplanted mouse endometriosis model, lenvatinib (VEGFR inhibitor) treatment","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — EV-mediated transfer functional assay, transcription factor regulation study, in vivo model; single lab, multiple methods","pmids":["33004630"],"is_preprint":false},{"year":2009,"finding":"TGF-β1 induces VEGF-C expression in gastric cancer cells via Smad-dependent pathway: phospho-Smad3 in the nucleus directly binds to the VEGFC promoter. Additionally, a Smad-independent AKT pathway also activates VEGF-C expression in response to TGF-β1 in a cell-line-dependent manner.","method":"Western blot for Smad2/3 phosphorylation, EMSA (electrophoretic mobility shift assay) for Smad3 binding to VEGFC promoter, AKT pathway inhibition, TGF-β1 inhibitor treatment, lymphatic tube forming assay, xenograft mouse model","journal":"BMC cancer","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — EMSA for direct promoter binding plus functional in vitro and in vivo assays; single lab","pmids":["31409309"],"is_preprint":false},{"year":2002,"finding":"Cyclic pressure (60/20 mmHg sinusoidal) selectively induces VEGF-C transcription in human umbilical vein endothelial cells, and VEGF-C mediates the cyclic pressure-induced endothelial cell proliferation response.","method":"Affymetrix GeneChip transcriptional profiling, cyclic pressure apparatus, VEGF-C-mediated proliferation assay","journal":"Physiological genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — transcriptomic identification plus functional proliferation assay demonstrating mediation; single lab","pmids":["12388793"],"is_preprint":false},{"year":2005,"finding":"Androgen ablation in LNCaP prostate cancer cells upregulates VEGF-C transcription. The mechanism involves downregulation of the IGF-IR pathway, with FOXO-1 (activated by SIRT-1) identified as the downstream transcriptional mediator of VEGF-C upregulation.","method":"Androgen withdrawal in LNCaP cells, FOXO-1 manipulation, SIRT-1 involvement analysis, VEGF-C mRNA measurement","journal":"Oncogene","confidence":"Low","confidence_rationale":"Tier 3 / Weak — pathway inhibition and transcription factor identification; single lab, limited mechanistic detail in abstract","pmids":["15897888"],"is_preprint":false},{"year":2019,"finding":"Hes1 transcription factor directly binds and positively regulates VEGF-C gene expression (a rare positive target of Hes1). VEGF-C upregulation by Hes1 contributes to attenuation of TLR upstream signaling by reducing WDFY1 expression, thereby suppressing type I IFN production.","method":"Genome-wide Hes1 occupancy (ChIP), VEGF-C expression analysis after Hes1 manipulation, Hes1-deficient mice, antiviral and autoimmune phenotype analysis","journal":"The Journal of experimental medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP for direct promoter binding, genetic knockout in mice; single lab, two methods","pmids":["31015298"],"is_preprint":false},{"year":2006,"finding":"VEGF-A upregulates VEGF-C mRNA expression and secretion in human retinal pigment epithelial (RPE) cells, and VEGF-A also stimulates VEGFR-3 mRNA expression. VEGF-C acts synergistically with VEGF-A to promote in vitro tube formation by choroidal endothelial cells.","method":"RT-PCR, western blotting, ELISA, Matrigel tube formation assay in choroidal endothelial cells, VEGF-A treatment of RPE cells","journal":"The British journal of ophthalmology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct transcriptional and secretion measurements, functional tube formation assay; single lab, multiple methods","pmids":["16687456"],"is_preprint":false},{"year":2014,"finding":"RhoGDI2 upregulates VEGF-C expression in gastric cancer cells, positively correlating with VEGF-C in human gastric tumor tissues. VEGF-C depletion suppresses RhoGDI2-induced gastric cancer metastasis and restores sensitivity to cisplatin-induced apoptosis. RhoGDI2 positively regulates Rac1 activity, and Rac1 inhibition suppresses RhoGDI2-induced VEGF-C expression.","method":"VEGF-C depletion (siRNA) in RhoGDI2-overexpressing cells, Rac1 inhibition, in vitro invasion assays, in vivo tumor metastasis models, immunohistochemistry of human tumor tissues","journal":"International journal of cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with defined pathway (RhoGDI2→Rac1→VEGF-C), in vivo validation; single lab","pmids":["24585459"],"is_preprint":false},{"year":2024,"finding":"VEGF-C overexpression (via AAV-mVEGF-C) in mouse brain increases CSF drainage to deep cervical lymph nodes by enhancing meningeal lymphatic vessel growth. VEGF-C prophylaxis reduces ischemic stroke injury and inflammation through a mechanism dependent on lymphatic drainage (neuroprotection was lost upon cauterization of deep cervical lymph node afferent lymphatics) and is associated with increased BDNF signaling and mitigated microglia-mediated inflammation.","method":"Intracerebrospinal AAV injection, single nuclei RNA sequencing, CSF drainage assay, mouse ischemic stroke model, cauterization of afferent lymphatics (epistasis), motor performance testing, immunohistochemistry","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Moderate — AAV-mediated gain-of-function with lymphatic cauterization epistasis establishing mechanism, multiple orthogonal readouts; rigorous single study","pmids":["38442272"],"is_preprint":false},{"year":2019,"finding":"VEGF-C/VEGFR3 signaling is required for wound lymphangiogenesis and anti-VEGF-C lymphangiogenesis blockade mediates a pro-inflammatory wound microenvironment and delayed wound closure. Targeted delivery of VEGF-C (F8-VEGF-C antibody fusion) to EDA-fibronectin-expressing remodeling tissue promotes wound healing, induces lymphangiogenesis, and reduces tissue inflammation in diabetic mice.","method":"Transgenic mice with increased/absent lymphatic vessels, VEGF-C/VEGFR3 pathway blocking, F8-VEGF-C fusion protein in db/db diabetic wound model, wound closure measurements, immunohistochemistry","journal":"Cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic gain/loss-of-function plus targeted delivery experiment; single lab, multiple models","pmids":["36766814"],"is_preprint":false}],"current_model":"VEGF-C is a secreted growth factor that is produced as an inactive precursor and activated by stepwise proteolytic processing (by plasmin and other proteases) to release the VEGF homology domain; partially processed forms selectively activate VEGFR-3 while fully mature VEGF-C also activates VEGFR-2 and binds neuropilin-2 (NRP2) via a cryptic C-terminal motif, forming a VEGFR-3/NRP2 signaling complex on lymphatic endothelial cells to drive lymphangiogenesis; VEGF-C is stored in platelet alpha-granules and released upon hemostasis to couple clotting with lymphatic regeneration; it is required for intestinal lacteal maintenance, fetal liver erythropoiesis, bone marrow perivascular niche integrity, and meningeal lymphatic drainage; its transcription is regulated by C/EBP-δ/HIF-1α in lymphatic endothelium, by Smad3 and AKT downstream of TGF-β1, by Hes1, and its mRNA is stabilized or degraded via the OTUD3/ZFP36/3'-UTR decay axis; autocrine VEGF-C/VEGFR-2 signaling also sustains glioblastoma cell survival and podocyte survival, and VEGF-C acts through NRP2 to non-canonically activate GLI signaling in breast cancer paracrine metastasis."},"narrative":{"mechanistic_narrative":"VEGF-C is a secreted lymphangiogenic growth factor that drives the proliferation of lymphatic endothelial cells and the formation of new lymphatic vessels, an activity not shared by VEGF-A or PlGF [PMID:9245515]. It is synthesized as an inactive precursor and activated by stepwise proteolytic processing: progressive cleavage releases the VEGF homology domain and generates forms with increasing potency, with partially processed material preferentially activating VEGFR-3 and only fully mature VEGF-C engaging VEGFR-2 [PMID:9233800, PMID:9012504]. Plasmin, and during hemostasis also thrombin, performs this maturation by removing N- and C-terminal propeptides to enhance receptor binding and cross-linking [PMID:12963694, PMID:31562136]. Proteolysis additionally exposes a cryptic C-terminal motif that binds the b1 domain of neuropilin-2 (NRP2), which co-internalizes with and co-precipitates with VEGFR-3 to form an active signaling complex on lymphatic endothelial cells [PMID:16816121, PMID:25752543]. VEGF-C is stored in platelet alpha-granules and released upon platelet activation, coupling hemostasis to wound lymphangiogenesis [PMID:9684805, PMID:31562136]. Beyond development, VEGF-C is required in adult tissues to maintain intestinal lacteals and dietary lipid absorption [PMID:26459520], to support fetal liver erythropoiesis [PMID:27343251], and to preserve the bone marrow perivascular niche and hematopoietic recovery [PMID:32842144]; engineered VEGF-C delivery enhances meningeal lymphatic CSF drainage and is neuroprotective after stroke [PMID:38442272]. VEGF-C transcription is controlled by C/EBP-δ acting with HIF-1α in lymphatic endothelium [PMID:21666710], by Smad3 and AKT downstream of TGF-β1 [PMID:31409309], and by Hes1 [PMID:31015298], while its mRNA is degraded through an OTUD3–ZFP36–3'-UTR decay axis [PMID:34853315]. In disease, VEGF-C supports autocrine VEGFR-2-driven survival in glioblastoma [PMID:29939339] and acts through NRP2 to non-canonically activate GLI signaling in breast cancer paracrine metastasis [PMID:33299122].","teleology":[{"year":1996,"claim":"Establishing that a secreted factor preferentially activates the lymphatic-associated receptor VEGFR-3 and is expressed near sites of lymphatic sprouting set up VEGF-C as a candidate lymphatic morphogen.","evidence":"Recombinant protein receptor autophosphorylation assays plus in situ hybridization in mouse embryos","pmids":["9012504"],"confidence":"High","gaps":["Did not establish whether VEGF-C is required for lymphatic development in vivo","Processing requirements for receptor selectivity not yet defined"]},{"year":1997,"claim":"Defining stepwise proteolytic processing as the switch governing receptor selectivity answered how a single ligand can engage VEGFR-3 versus VEGFR-2, and direct lymphangiogenesis assays established VEGF-C as the first lymphangiogenic growth factor.","evidence":"Recombinant protein binding/autophosphorylation kinetics and in vivo chorioallantoic membrane lymphangiogenesis assay with growth factor controls","pmids":["9233800","9245515"],"confidence":"High","gaps":["Identity of the activating protease(s) not yet determined","Structural basis of receptor engagement not resolved"]},{"year":2003,"claim":"Identifying plasmin as a protease that cleaves both propeptides connected VEGF-C activation to a defined enzymatic event rather than an unknown maturation step.","evidence":"In vitro protease cleavage and receptor binding/cross-linking assays comparing full-length and processed VEGF-C","pmids":["12963694"],"confidence":"High","gaps":["Did not address physiological context in which plasmin activation occurs","Other contributing proteases not excluded"]},{"year":2006,"claim":"Showing direct VEGF-C binding to neuropilin-2 and NRP2 co-internalization with VEGFR-3 placed NRP2 as a partner within an active lymphatic signaling complex rather than a passive co-receptor.","evidence":"In vitro binding, domain mapping, co-immunoprecipitation, and co-internalization imaging in lymphatic endothelial cells","pmids":["16816121"],"confidence":"High","gaps":["Structural determinant of the VEGF-C/NRP2 interaction not resolved","Functional consequence of complex internalization for signaling output not quantified"]},{"year":2015,"claim":"A crystal structure of the VEGF-C C-terminus bound to NRP2 explained how proteolysis unmasks a cryptic NRP2-binding motif and how a secreted splice form can antagonize it.","evidence":"X-ray crystallography with in vitro binding and cellular signaling assays","pmids":["25752543"],"confidence":"High","gaps":["In vivo role of the s9Nrp2 antagonist not established","Structure does not capture the full VEGFR-3/NRP2 ternary complex"]},{"year":2019,"claim":"Demonstrating that platelet-stored VEGF-C is activated by hemostatic proteases and is the dominant driver of wound lymphangiogenesis linked clotting directly to lymphatic regeneration.","evidence":"In vitro thrombin/plasmin cleavage assays plus platelet-specific Vegfc deletion in tail-wounding and full-thickness excision mouse models","pmids":["31562136","9684805","36766814"],"confidence":"High","gaps":["Relative contributions of thrombin versus plasmin in vivo not quantified","Mechanism linking inflammation resolution to VEGF-C-driven lymphatics incomplete"]},{"year":2016,"claim":"Conditional and temporally controlled Vegfc deletion revealed non-lymphangiogenic adult and developmental roles—maintaining intestinal lacteals and enabling fetal liver erythropoiesis—broadening VEGF-C beyond a purely lymphatic factor.","evidence":"Conditional Vegfc knockout in adult and embryonic mice with cellular, lipid-absorption, and hematopoietic readouts","pmids":["26459520","27343251"],"confidence":"High","gaps":["Receptor and cellular target mediating the erythropoietic effect not fully defined","Whether lacteal maintenance and erythropoiesis share a common mechanism unknown"]},{"year":2020,"claim":"Loss- and gain-of-function studies established VEGF-C as a maintenance factor for the bone marrow perivascular niche, with exogenous delivery accelerating hematopoietic recovery.","evidence":"Endothelial- and LepR+-specific Vegfc deletion plus AAV-VEGF-C rescue in an irradiation/transplant model","pmids":["32842144"],"confidence":"High","gaps":["Direct receptor mediating niche maintenance not pinned down","Identity of induced hematopoietic regenerative factors only partially defined"]},{"year":2021,"claim":"Reconstitution of an OTUD3–ZFP36 axis acting on the VEGF-C 3'-UTR identified a post-transcriptional control layer governing VEGF-C abundance and lymphatic metastasis.","evidence":"Co-IP, ubiquitination and RNA decay assays, 3'-UTR binding assays, and an in vivo metastasis model","pmids":["34853315","21666710","31409309","31015298"],"confidence":"High","gaps":["Generality of this decay axis across non-cancer tissues not established","Interplay between transcriptional and post-transcriptional control not integrated"]},{"year":2024,"claim":"AAV-driven VEGF-C overexpression coupled with lymphatic cauterization epistasis demonstrated that VEGF-C-enhanced meningeal lymphatic drainage is neuroprotective after stroke, extending its physiology to the CNS.","evidence":"Intra-CSF AAV-VEGF-C, single-nuclei RNA-seq, CSF drainage assays, and stroke model with afferent lymphatic cauterization","pmids":["38442272"],"confidence":"High","gaps":["Endogenous VEGF-C requirement for meningeal lymphatics not tested by loss-of-function","Direct molecular link between drainage and BDNF/microglial changes incomplete"]},{"year":null,"claim":"How the distinct VEGF-C receptor/co-receptor configurations (VEGFR-3, VEGFR-2, VEGFR-1, NRP2) are selected in different cell types to produce lymphangiogenic versus autocrine survival versus non-canonical GLI outputs remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["Cell-context determinants of receptor choice not defined","Conflicting receptor usage in podocytes (VEGFR-1 vs VEGFR-2) unreconciled","Mechanism of NRP2-to-GLI signal transduction unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[0,1,2,3]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,4,5]}],"localization":[{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[0,1,6]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[6,22]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,4,5]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[2,8,13]},{"term_id":"R-HSA-1500931","term_label":"Cell-Cell 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Mature VEGF-C binds VEGFR-3 (Kd ~135 pM) and VEGFR-2 (Kd ~410 pM), activates both receptors, increases vascular permeability, and stimulates endothelial cell migration and proliferation. Unlike other PDGF/VEGF family members, mature VEGF-C forms predominantly non-covalent homodimers.\",\n      \"method\": \"Recombinant protein production in yeast, receptor binding assays, receptor autophosphorylation assays, vascular permeability assays, endothelial cell migration and proliferation assays, biochemical characterization of processing intermediates\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with recombinant proteins, binding kinetics, functional receptor activation assays, multiple orthogonal methods in a single rigorous study\",\n      \"pmids\": [\"9233800\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"Mouse VEGF-C is secreted as VEGFR-3 (Flt4)-binding polypeptides of 30–32 kDa and 22–23 kDa that preferentially stimulate VEGFR-3 autophosphorylation over VEGFR-2. In situ hybridization shows VEGF-C mRNA in mesenchymal cells near regions where lymphatic vessels sprout from embryonic veins, consistent with a paracrine role in lymphatic vessel development.\",\n      \"method\": \"Recombinant protein production, receptor autophosphorylation assay (VEGFR-3 vs VEGFR-2), in situ hybridization in mouse embryos\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — receptor activation assays plus spatial expression mapping, replicated across multiple labs\",\n      \"pmids\": [\"9012504\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"VEGF-C is the first identified lymphangiogenic growth factor; it induces proliferation of lymphatic endothelial cells and formation of new lymphatic sinuses in the avian chorioallantoic membrane (CAM). VEGF, by contrast, is angiogenic but not lymphangiogenic in this model, and PlGF has neither activity.\",\n      \"method\": \"In vivo CAM assay with microinjection, immunohistochemistry, in situ hybridization for VEGFR-2 and VEGFR-3\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct in vivo lymphangiogenesis assay with comparative growth factor controls, independently replicated\",\n      \"pmids\": [\"9245515\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"The serine protease plasmin cleaves both NH2- and COOH-terminal propeptides from the VEGF homology domain of VEGF-C (and VEGF-D), generating a mature form with greatly enhanced binding and cross-linking of VEGFR-2 and VEGFR-3 compared to full-length material. Plasmin thereby activates VEGF-C.\",\n      \"method\": \"In vitro protease cleavage assay, receptor binding and cross-linking assays with full-length versus plasmin-processed VEGF-C\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct in vitro enzymatic reconstitution with biochemical characterization of cleavage products and receptor binding, independently replicated concept\",\n      \"pmids\": [\"12963694\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"VEGF-C and VEGF-D directly interact with neuropilin-2 (NP2): VEGF-C binds NP2 in a heparin-independent manner while VEGF-D binding is heparin-dependent. The domains of VEGF-C and NP2 required for this interaction were mapped. NP2 co-internalizes with VEGFR-3 into endocytic vesicles of lymphatic endothelial cells upon VEGF-C or VEGF-D stimulation, and NP2 co-precipitates with VEGFR-3, indicating that NP2 participates in an active signaling complex with VEGFR-3.\",\n      \"method\": \"In vitro binding studies, domain mapping, co-immunoprecipitation, co-internalization imaging in lymphatic endothelial cells\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal binding studies, co-IP, and co-internalization imaging; multiple orthogonal methods in single study\",\n      \"pmids\": [\"16816121\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Proteolytic maturation of VEGF-C releases a cryptic Nrp2-binding motif at the C-terminus, allowing specific engagement with the b1 domain of Nrp2. Crystal structure of the VEGF-C C-terminus in complex with Nrp2 ligand-binding domains was determined. The endogenous secreted splice form s9Nrp2 forms a stable dimer and potently inhibits VEGF-C/Nrp2 binding and downstream cellular signaling.\",\n      \"method\": \"X-ray crystallography, in vitro binding assays, cellular signaling assays\",\n      \"journal\": \"Structure (London, England : 1993)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure with functional binding validation and cellular assays in a single rigorous study\",\n      \"pmids\": [\"25752543\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"VEGF-C protein is stored in platelet alpha-granules and is released from platelets upon activation, together with beta-thromboglobulin. VEGF-C mRNA in peripheral blood is restricted to platelets and T cells.\",\n      \"method\": \"VEGF-C mRNA detection in blood cell fractions, VEGF-C protein release assay from activated platelets, co-release with beta-thromboglobulin as alpha-granule marker\",\n      \"journal\": \"Thrombosis and haemostasis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell fractionation, protein release assay, co-release with granule marker; single lab, two orthogonal methods\",\n      \"pmids\": [\"9684805\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"VEGF-C is cleaved and activated by thrombin and plasmin generated during hemostasis. Platelets accelerate lymphatic growth after injury in vivo, and this platelet-enhanced lymphangiogenesis depends on platelet-derived VEGF-C (but not VEGF-D). VEGF-C, but not VEGF-D, is also the dominant factor for lymphangiogenesis after full-thickness skin excision.\",\n      \"method\": \"In vitro protease cleavage assays, tail-wounding mouse model, genetic studies with platelet-specific VEGF-C deletion, full-thickness excision wound model\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro biochemical reconstitution plus in vivo genetic studies with two wound models and multiple orthogonal approaches\",\n      \"pmids\": [\"31562136\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"VEGF-C is required for maintenance of intestinal lymphatic vessels (lacteals) in adult mice. VEGF-C is expressed by smooth muscle cells adjacent to lacteals. Deletion of Vegfc in adult mice causes gradual atrophy of intestinal lymphatics, leading to defective lipid absorption, increased fecal excretion of dietary cholesterol and fatty acids, and resistance to diet-induced obesity.\",\n      \"method\": \"Conditional Vegfc gene deletion in adult mice, immunohistochemistry for lymphatic markers, lipid absorption assays, dietary challenge experiments\",\n      \"journal\": \"EMBO molecular medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional knockout with multiple phenotypic readouts and cell-of-origin identification; rigorous genetic study\",\n      \"pmids\": [\"26459520\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"VEGF-C signaling via VEGFR-2 works synergistically with VEGF-A to enhance vascular bed formation. VEGF-C binding to VEGFR-3 sequesters VEGF-C away from VEGFR-2, thereby regulating VEGFR-2 signaling. In VEGFR-3-deficient embryos, excess VEGF-C signals through VEGFR-2, causing disturbed vasculogenesis and suppressed hematopoiesis.\",\n      \"method\": \"Para-aortic splanchnopleural mesoderm (P-Sp) coculture with OP9 stromal cells, soluble receptor competitive inhibition, VEGFR-3-deficient embryo explants\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and pharmacological epistasis in an ex vivo embryo coculture system; single lab with multiple experimental conditions\",\n      \"pmids\": [\"11090062\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"FGF-2-induced lymphangiogenesis requires VEGFR-3 signaling; a VEGFR-3-neutralizing antibody markedly inhibits FGF-2-induced lymphangiogenesis. VEGFR-3-mediated signaling is a prerequisite for lymphatic tip cell formation in both FGF-2- and VEGF-C-induced lymphangiogenesis. FGFR-1 expressed in lymphatic endothelial cells is the receptor mediating FGF-2-induced lymphangiogenesis.\",\n      \"method\": \"Mouse corneal lymphangiogenesis assay, VEGFR-3 neutralizing antibody epistasis, tumor co-implantation model, in vitro lymphatic endothelial cell assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo epistasis with neutralizing antibody, multiple tumor and corneal models; single lab\",\n      \"pmids\": [\"22967508\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Hypoxia reduces VEGF-C cap-dependent translation via upregulation of hypophosphorylated 4E-BP1, but induces VEGF-C translation through an internal ribosome entry site (IRES)-dependent mechanism that is independent of HIF-1α. This IRES-dependent VEGF-C translation is higher in lymph node metastases than in primary tumors.\",\n      \"method\": \"IRES reporter assays, 4E-BP1 manipulation, HIF-1α knockdown, comparison of primary tumor vs. lymph node metastasis samples\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic dissection using reporter assays and genetic knockdown; multiple orthogonal approaches in single lab\",\n      \"pmids\": [\"24388748\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"VEGF-C regulates capillary stabilization by controlling PDGF-B expression via VEGFR-3 in a paracrine mode during neovascularization. Blockade of VEGFR-3 inhibited FGF-2-mediated limb salvage and caused capillary dilation with mural cell dissociation, and VEGF-C and PDGF-B were mutually dependent (blocking VEGFR-3 decreased PDGF-B expression, and blocking PDGF-BB decreased VEGF-C expression).\",\n      \"method\": \"Murine ischemic hindlimb model, VEGFR-3 neutralizing antibody AFL-4, PDGF-BB blocking antibody, immunohistochemistry\",\n      \"journal\": \"American journal of physiology. Heart and circulatory physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo epistasis with neutralizing antibodies and multiple phenotypic readouts; single lab\",\n      \"pmids\": [\"19734356\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"VEGF-C is required for the transition to fetal liver erythropoiesis. Embryonic Vegfc deletion causes defective fetal erythropoiesis with anemia and absence of enucleated red blood cells, reduced macrophages and erythroid cells in fetal liver due to decreased proliferation and increased apoptosis, decreased α4-integrin on erythro-myeloid progenitors (EMPs), and impaired EMP colonization of the fetal liver. Vegfc deletion from E10.5 onward or in adults does not affect definitive hematopoiesis.\",\n      \"method\": \"Conditional Vegfc deletion at E7.5, flow cytometry, immunohistochemistry, blood cell analysis, integrin expression assays\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional genetic knockout with temporally controlled deletion, multiple cellular and molecular readouts, defines a novel non-lymphangiogenic function\",\n      \"pmids\": [\"27343251\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"VEGF-C maintains the integrity of the bone marrow perivascular niche. Global and conditional deletion of Vegfc from endothelial or LepR+ cells disrupts the BM perivascular niche and delays hematopoietic recovery after irradiation by decreasing endothelial proliferation and LepR+ cell regeneration. Exogenous AAV-delivered VEGF-C improves hematopoietic recovery by accelerating endothelial and LepR+ cell regeneration and increasing expression of hematopoietic regenerative factors.\",\n      \"method\": \"Conditional Vegfc gene deletion from endothelial and LepR+ cells, irradiation/transplantation model, AAV-mediated VEGF-C delivery, immunohistochemistry, flow cytometry\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional knockout with cell-type specificity, rescue experiment with exogenous VEGF-C, multiple orthogonal readouts\",\n      \"pmids\": [\"32842144\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"VEGF-C acts as both a paracrine and autocrine pro-survival cytokine in glioblastoma, activating VEGFR-2 in an autocrine manner to promote tumor cell survival and sustain VEGFR2 activation that is bevacizumab-resistant. Targeting VEGF-C expression in vivo impairs tumor growth more effectively than bevacizumab treatment.\",\n      \"method\": \"RNA interference, exogenous ligand treatment, proximity ligation assay for VEGF-C/VEGFR2 interaction in tumor specimens, patient-derived xenograft in vivo experiments, matched pre/post-bevacizumab treatment cohort analysis\",\n      \"journal\": \"Neuro-oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNAi knockdown with functional readouts, proximity ligation assay for direct interaction, in vivo xenograft; single lab\",\n      \"pmids\": [\"29939339\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"VEGF-C secreted by breast cancer cells that have undergone EMT promotes paracrine non-canonical GLI signaling activation in neighboring epithelial breast cancer cells via NRP2. Inhibiting VEGF-C in EMT cells or knocking down NRP2 in epithelial cancer cells disrupts the aggressive phenotypes (proliferation, migration, invasion, metastasis) imparted by EMT cells.\",\n      \"method\": \"VEGF-C knockdown in EMT cells, NRP2 knockdown in epithelial cells, co-culture systems, in vivo metastasis models, TCGA/GEO dataset correlation\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined pathway placement (VEGF-C/NRP2/GLI axis), in vivo validation; single lab\",\n      \"pmids\": [\"33299122\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"C/EBP-δ transcription factor regulates VEGF-C and VEGFR-3 expression specifically in lymphatic endothelial cells (LECs). Genetic deletion of C/EBP-δ in mice dramatically reduces VEGF-C and VEGFR-3 in LECs, reduces lymphangiogenesis and pulmonary metastases. Forced VEGF-C expression (but not recombinant VEGF-C protein) rescues C/EBP-δ knockdown-induced LEC apoptosis, demonstrating autocrine VEGF-C signaling is essential for LEC survival. C/EBP-δ-induced VEGF-C expression requires HIF-1α.\",\n      \"method\": \"C/EBP-δ knockout mice, forced expression/knockdown in cultured LECs, HIF-1α blocking, rescue experiments with VEGF-C plasmid vs. recombinant protein\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockout in mice, forced expression/knockdown, mechanistic dissection with HIF-1α; single lab, multiple methods\",\n      \"pmids\": [\"21666710\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Autocrine VEGF-C (along with VEGF-A) in human podocytes is important for podocyte survival via VEGFR-2-mediated activation of PI3K/AKT (anti-apoptotic) and suppression of p38MAPK (pro-apoptotic). Exogenous VEGF-C can reverse the effect of VEGF-A neutralization, indicating functional overlap and complementarity between autocrine VEGF-A and VEGF-C in podocytes.\",\n      \"method\": \"siRNA knockdown of VEGF-A and VEGF-C in human podocytes, bevacizumab treatment, VEGFR-2/-3 tyrosine kinase inhibitor, phospho-AKT and p38MAPK western blotting, apoptosis assays\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA loss-of-function with defined signaling readouts and rescue experiments; single lab\",\n      \"pmids\": [\"19828679\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"VEGF-C acts as an autocrine survival factor in cultured human podocytes via VEGFR-1 (not VEGFR-3, as VEGF-C did not autophosphorylate VEGFR-3 in these cells despite doing so in HMVECs). VEGF-C reduces intracellular calcium and cytotoxicity, reduces MAPK phosphorylation, and has no effect on Akt phosphorylation in podocytes.\",\n      \"method\": \"VEGF-C treatment of conditionally immortalized human podocytes, VEGFR-3 kinase inhibitor (MAZ51), SU-5416 (VEGFR-1-specific concentrations), immunoprecipitation for VEGFR-3 autophosphorylation, calcium imaging, cytotoxicity assays\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological inhibition with receptor-specific reagents and receptor immunoprecipitation; single lab, multiple assays\",\n      \"pmids\": [\"16525158\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"VEGF-C drives bone lymphatic vessel formation via VEGFR-3 (but not VEGFR-2) signaling, as inhibition of VEGFR-3 but not VEGFR-2 prevented formation of bone lymphatics in Osx-tTA;TetO-Vegfc mice. VEGF-C overexpression in bone also promotes bone loss via increased osteoclast numbers, which is attenuated by an osteoclast inhibitor.\",\n      \"method\": \"Double-transgenic mouse model (Osx-tTA;TetO-Vegfc), VEGFR-selective antibody blockade, radiological and histological bone analysis, osteoclast inhibitor treatment\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic gain-of-function with receptor-selective inhibition epistasis; single lab\",\n      \"pmids\": [\"29620526\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"OTUD3 deubiquitinase stabilizes ZFP36 by inhibiting FBXW7-mediated K48-linked polyubiquitination of ZFP36. ZFP36 binds the VEGF-C 3'-UTR and recruits the RNA-degrading complex to induce rapid VEGF-C mRNA decay. Nicotine downregulates OTUD3, leading to ZFP36 destabilization and increased VEGF-C production and lymphatic metastasis.\",\n      \"method\": \"Co-immunoprecipitation of OTUD3-ZFP36 interaction, ubiquitination assays, RNA decay assays, ZFP36-VEGF-C 3'-UTR binding assays, OTUD3 knockdown/overexpression in cancer cells, in vivo metastasis model\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — biochemical reconstitution of the ubiquitination pathway, RNA decay assays, direct 3'-UTR binding, in vivo validation; multiple orthogonal methods\",\n      \"pmids\": [\"34853315\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Functional VEGF-C is transported by extracellular vesicles (EVs) from endometriotic cells to lymphatic endothelial cells, enhancing their lymphangiogenic ability. VEGF-C expression in endometriotic cells is negatively regulated by COUP-TFII, and proinflammatory cytokines increase VEGF-C by suppressing COUP-TFII levels.\",\n      \"method\": \"EV isolation and functional transfer assay, COUP-TFII knockdown/overexpression, cytokine treatment, autotransplanted mouse endometriosis model, lenvatinib (VEGFR inhibitor) treatment\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — EV-mediated transfer functional assay, transcription factor regulation study, in vivo model; single lab, multiple methods\",\n      \"pmids\": [\"33004630\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"TGF-β1 induces VEGF-C expression in gastric cancer cells via Smad-dependent pathway: phospho-Smad3 in the nucleus directly binds to the VEGFC promoter. Additionally, a Smad-independent AKT pathway also activates VEGF-C expression in response to TGF-β1 in a cell-line-dependent manner.\",\n      \"method\": \"Western blot for Smad2/3 phosphorylation, EMSA (electrophoretic mobility shift assay) for Smad3 binding to VEGFC promoter, AKT pathway inhibition, TGF-β1 inhibitor treatment, lymphatic tube forming assay, xenograft mouse model\",\n      \"journal\": \"BMC cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — EMSA for direct promoter binding plus functional in vitro and in vivo assays; single lab\",\n      \"pmids\": [\"31409309\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Cyclic pressure (60/20 mmHg sinusoidal) selectively induces VEGF-C transcription in human umbilical vein endothelial cells, and VEGF-C mediates the cyclic pressure-induced endothelial cell proliferation response.\",\n      \"method\": \"Affymetrix GeneChip transcriptional profiling, cyclic pressure apparatus, VEGF-C-mediated proliferation assay\",\n      \"journal\": \"Physiological genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — transcriptomic identification plus functional proliferation assay demonstrating mediation; single lab\",\n      \"pmids\": [\"12388793\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Androgen ablation in LNCaP prostate cancer cells upregulates VEGF-C transcription. The mechanism involves downregulation of the IGF-IR pathway, with FOXO-1 (activated by SIRT-1) identified as the downstream transcriptional mediator of VEGF-C upregulation.\",\n      \"method\": \"Androgen withdrawal in LNCaP cells, FOXO-1 manipulation, SIRT-1 involvement analysis, VEGF-C mRNA measurement\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — pathway inhibition and transcription factor identification; single lab, limited mechanistic detail in abstract\",\n      \"pmids\": [\"15897888\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Hes1 transcription factor directly binds and positively regulates VEGF-C gene expression (a rare positive target of Hes1). VEGF-C upregulation by Hes1 contributes to attenuation of TLR upstream signaling by reducing WDFY1 expression, thereby suppressing type I IFN production.\",\n      \"method\": \"Genome-wide Hes1 occupancy (ChIP), VEGF-C expression analysis after Hes1 manipulation, Hes1-deficient mice, antiviral and autoimmune phenotype analysis\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP for direct promoter binding, genetic knockout in mice; single lab, two methods\",\n      \"pmids\": [\"31015298\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"VEGF-A upregulates VEGF-C mRNA expression and secretion in human retinal pigment epithelial (RPE) cells, and VEGF-A also stimulates VEGFR-3 mRNA expression. VEGF-C acts synergistically with VEGF-A to promote in vitro tube formation by choroidal endothelial cells.\",\n      \"method\": \"RT-PCR, western blotting, ELISA, Matrigel tube formation assay in choroidal endothelial cells, VEGF-A treatment of RPE cells\",\n      \"journal\": \"The British journal of ophthalmology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct transcriptional and secretion measurements, functional tube formation assay; single lab, multiple methods\",\n      \"pmids\": [\"16687456\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RhoGDI2 upregulates VEGF-C expression in gastric cancer cells, positively correlating with VEGF-C in human gastric tumor tissues. VEGF-C depletion suppresses RhoGDI2-induced gastric cancer metastasis and restores sensitivity to cisplatin-induced apoptosis. RhoGDI2 positively regulates Rac1 activity, and Rac1 inhibition suppresses RhoGDI2-induced VEGF-C expression.\",\n      \"method\": \"VEGF-C depletion (siRNA) in RhoGDI2-overexpressing cells, Rac1 inhibition, in vitro invasion assays, in vivo tumor metastasis models, immunohistochemistry of human tumor tissues\",\n      \"journal\": \"International journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with defined pathway (RhoGDI2→Rac1→VEGF-C), in vivo validation; single lab\",\n      \"pmids\": [\"24585459\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"VEGF-C overexpression (via AAV-mVEGF-C) in mouse brain increases CSF drainage to deep cervical lymph nodes by enhancing meningeal lymphatic vessel growth. VEGF-C prophylaxis reduces ischemic stroke injury and inflammation through a mechanism dependent on lymphatic drainage (neuroprotection was lost upon cauterization of deep cervical lymph node afferent lymphatics) and is associated with increased BDNF signaling and mitigated microglia-mediated inflammation.\",\n      \"method\": \"Intracerebrospinal AAV injection, single nuclei RNA sequencing, CSF drainage assay, mouse ischemic stroke model, cauterization of afferent lymphatics (epistasis), motor performance testing, immunohistochemistry\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — AAV-mediated gain-of-function with lymphatic cauterization epistasis establishing mechanism, multiple orthogonal readouts; rigorous single study\",\n      \"pmids\": [\"38442272\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"VEGF-C/VEGFR3 signaling is required for wound lymphangiogenesis and anti-VEGF-C lymphangiogenesis blockade mediates a pro-inflammatory wound microenvironment and delayed wound closure. Targeted delivery of VEGF-C (F8-VEGF-C antibody fusion) to EDA-fibronectin-expressing remodeling tissue promotes wound healing, induces lymphangiogenesis, and reduces tissue inflammation in diabetic mice.\",\n      \"method\": \"Transgenic mice with increased/absent lymphatic vessels, VEGF-C/VEGFR3 pathway blocking, F8-VEGF-C fusion protein in db/db diabetic wound model, wound closure measurements, immunohistochemistry\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic gain/loss-of-function plus targeted delivery experiment; single lab, multiple models\",\n      \"pmids\": [\"36766814\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"VEGF-C is a secreted growth factor that is produced as an inactive precursor and activated by stepwise proteolytic processing (by plasmin and other proteases) to release the VEGF homology domain; partially processed forms selectively activate VEGFR-3 while fully mature VEGF-C also activates VEGFR-2 and binds neuropilin-2 (NRP2) via a cryptic C-terminal motif, forming a VEGFR-3/NRP2 signaling complex on lymphatic endothelial cells to drive lymphangiogenesis; VEGF-C is stored in platelet alpha-granules and released upon hemostasis to couple clotting with lymphatic regeneration; it is required for intestinal lacteal maintenance, fetal liver erythropoiesis, bone marrow perivascular niche integrity, and meningeal lymphatic drainage; its transcription is regulated by C/EBP-δ/HIF-1α in lymphatic endothelium, by Smad3 and AKT downstream of TGF-β1, by Hes1, and its mRNA is stabilized or degraded via the OTUD3/ZFP36/3'-UTR decay axis; autocrine VEGF-C/VEGFR-2 signaling also sustains glioblastoma cell survival and podocyte survival, and VEGF-C acts through NRP2 to non-canonically activate GLI signaling in breast cancer paracrine metastasis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"VEGF-C is a secreted lymphangiogenic growth factor that drives the proliferation of lymphatic endothelial cells and the formation of new lymphatic vessels, an activity not shared by VEGF-A or PlGF [#2]. It is synthesized as an inactive precursor and activated by stepwise proteolytic processing: progressive cleavage releases the VEGF homology domain and generates forms with increasing potency, with partially processed material preferentially activating VEGFR-3 and only fully mature VEGF-C engaging VEGFR-2 [#0, #1]. Plasmin, and during hemostasis also thrombin, performs this maturation by removing N- and C-terminal propeptides to enhance receptor binding and cross-linking [#3, #7]. Proteolysis additionally exposes a cryptic C-terminal motif that binds the b1 domain of neuropilin-2 (NRP2), which co-internalizes with and co-precipitates with VEGFR-3 to form an active signaling complex on lymphatic endothelial cells [#4, #5]. VEGF-C is stored in platelet alpha-granules and released upon platelet activation, coupling hemostasis to wound lymphangiogenesis [#6, #7]. Beyond development, VEGF-C is required in adult tissues to maintain intestinal lacteals and dietary lipid absorption [#8], to support fetal liver erythropoiesis [#13], and to preserve the bone marrow perivascular niche and hematopoietic recovery [#14]; engineered VEGF-C delivery enhances meningeal lymphatic CSF drainage and is neuroprotective after stroke [#29]. VEGF-C transcription is controlled by C/EBP-\\u03b4 acting with HIF-1\\u03b1 in lymphatic endothelium [#17], by Smad3 and AKT downstream of TGF-\\u03b21 [#23], and by Hes1 [#26], while its mRNA is degraded through an OTUD3\\u2013ZFP36\\u20133'-UTR decay axis [#21]. In disease, VEGF-C supports autocrine VEGFR-2-driven survival in glioblastoma [#15] and acts through NRP2 to non-canonically activate GLI signaling in breast cancer paracrine metastasis [#16].\",\n  \"teleology\": [\n    {\n      \"year\": 1996,\n      \"claim\": \"Establishing that a secreted factor preferentially activates the lymphatic-associated receptor VEGFR-3 and is expressed near sites of lymphatic sprouting set up VEGF-C as a candidate lymphatic morphogen.\",\n      \"evidence\": \"Recombinant protein receptor autophosphorylation assays plus in situ hybridization in mouse embryos\",\n      \"pmids\": [\"9012504\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not establish whether VEGF-C is required for lymphatic development in vivo\", \"Processing requirements for receptor selectivity not yet defined\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Defining stepwise proteolytic processing as the switch governing receptor selectivity answered how a single ligand can engage VEGFR-3 versus VEGFR-2, and direct lymphangiogenesis assays established VEGF-C as the first lymphangiogenic growth factor.\",\n      \"evidence\": \"Recombinant protein binding/autophosphorylation kinetics and in vivo chorioallantoic membrane lymphangiogenesis assay with growth factor controls\",\n      \"pmids\": [\"9233800\", \"9245515\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the activating protease(s) not yet determined\", \"Structural basis of receptor engagement not resolved\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Identifying plasmin as a protease that cleaves both propeptides connected VEGF-C activation to a defined enzymatic event rather than an unknown maturation step.\",\n      \"evidence\": \"In vitro protease cleavage and receptor binding/cross-linking assays comparing full-length and processed VEGF-C\",\n      \"pmids\": [\"12963694\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address physiological context in which plasmin activation occurs\", \"Other contributing proteases not excluded\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Showing direct VEGF-C binding to neuropilin-2 and NRP2 co-internalization with VEGFR-3 placed NRP2 as a partner within an active lymphatic signaling complex rather than a passive co-receptor.\",\n      \"evidence\": \"In vitro binding, domain mapping, co-immunoprecipitation, and co-internalization imaging in lymphatic endothelial cells\",\n      \"pmids\": [\"16816121\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural determinant of the VEGF-C/NRP2 interaction not resolved\", \"Functional consequence of complex internalization for signaling output not quantified\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"A crystal structure of the VEGF-C C-terminus bound to NRP2 explained how proteolysis unmasks a cryptic NRP2-binding motif and how a secreted splice form can antagonize it.\",\n      \"evidence\": \"X-ray crystallography with in vitro binding and cellular signaling assays\",\n      \"pmids\": [\"25752543\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo role of the s9Nrp2 antagonist not established\", \"Structure does not capture the full VEGFR-3/NRP2 ternary complex\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Demonstrating that platelet-stored VEGF-C is activated by hemostatic proteases and is the dominant driver of wound lymphangiogenesis linked clotting directly to lymphatic regeneration.\",\n      \"evidence\": \"In vitro thrombin/plasmin cleavage assays plus platelet-specific Vegfc deletion in tail-wounding and full-thickness excision mouse models\",\n      \"pmids\": [\"31562136\", \"9684805\", \"36766814\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative contributions of thrombin versus plasmin in vivo not quantified\", \"Mechanism linking inflammation resolution to VEGF-C-driven lymphatics incomplete\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Conditional and temporally controlled Vegfc deletion revealed non-lymphangiogenic adult and developmental roles\\u2014maintaining intestinal lacteals and enabling fetal liver erythropoiesis\\u2014broadening VEGF-C beyond a purely lymphatic factor.\",\n      \"evidence\": \"Conditional Vegfc knockout in adult and embryonic mice with cellular, lipid-absorption, and hematopoietic readouts\",\n      \"pmids\": [\"26459520\", \"27343251\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor and cellular target mediating the erythropoietic effect not fully defined\", \"Whether lacteal maintenance and erythropoiesis share a common mechanism unknown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Loss- and gain-of-function studies established VEGF-C as a maintenance factor for the bone marrow perivascular niche, with exogenous delivery accelerating hematopoietic recovery.\",\n      \"evidence\": \"Endothelial- and LepR+-specific Vegfc deletion plus AAV-VEGF-C rescue in an irradiation/transplant model\",\n      \"pmids\": [\"32842144\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct receptor mediating niche maintenance not pinned down\", \"Identity of induced hematopoietic regenerative factors only partially defined\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Reconstitution of an OTUD3\\u2013ZFP36 axis acting on the VEGF-C 3'-UTR identified a post-transcriptional control layer governing VEGF-C abundance and lymphatic metastasis.\",\n      \"evidence\": \"Co-IP, ubiquitination and RNA decay assays, 3'-UTR binding assays, and an in vivo metastasis model\",\n      \"pmids\": [\"34853315\", \"21666710\", \"31409309\", \"31015298\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Generality of this decay axis across non-cancer tissues not established\", \"Interplay between transcriptional and post-transcriptional control not integrated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"AAV-driven VEGF-C overexpression coupled with lymphatic cauterization epistasis demonstrated that VEGF-C-enhanced meningeal lymphatic drainage is neuroprotective after stroke, extending its physiology to the CNS.\",\n      \"evidence\": \"Intra-CSF AAV-VEGF-C, single-nuclei RNA-seq, CSF drainage assays, and stroke model with afferent lymphatic cauterization\",\n      \"pmids\": [\"38442272\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Endogenous VEGF-C requirement for meningeal lymphatics not tested by loss-of-function\", \"Direct molecular link between drainage and BDNF/microglial changes incomplete\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the distinct VEGF-C receptor/co-receptor configurations (VEGFR-3, VEGFR-2, VEGFR-1, NRP2) are selected in different cell types to produce lymphangiogenic versus autocrine survival versus non-canonical GLI outputs remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Cell-context determinants of receptor choice not defined\", \"Conflicting receptor usage in podocytes (VEGFR-1 vs VEGFR-2) unreconciled\", \"Mechanism of NRP2-to-GLI signal transduction unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [0, 1, 2, 3]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 4, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0, 1, 6]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [6, 22]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 4, 5]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 8, 13]},\n      {\"term_id\": \"R-HSA-1500931\", \"supporting_discovery_ids\": [1, 12]}\n    ],\n    \"complexes\": [\"VEGFR-3/NRP2 signaling complex\"],\n    \"partners\": [\"FLT4\", \"KDR\", \"NRP2\", \"PLG\", \"F2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie","faith_supported":8,"faith_total":8,"faith_pct":100.0}}