{"gene":"CRIPTO","run_date":"2026-06-09T22:57:19","timeline":{"discoveries":[{"year":2001,"finding":"Cripto interacts with the type I receptor ALK4 via its conserved CFC motif. This interaction is necessary both for Nodal binding to the ALK4/ActRIIB receptor complex and for Smad2 activation by Nodal.","method":"Co-immunoprecipitation, receptor binding assays, Smad2 activation reporter assays in cell culture","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding and functional activation assays, replicated across multiple labs in subsequent papers","pmids":["11389842"],"is_preprint":false},{"year":2000,"finding":"Cripto protein is anchored to the cell membrane via a glycosylphosphatidylinositol (GPI) moiety, as demonstrated by biochemical characterization and immunofluorescence; GPI-linkage is proposed to be important for spatial specificity of cell-cell interactions.","method":"Biochemical fractionation, PI-PLC treatment, immunofluorescence","journal":"Mechanisms of development","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal biochemical methods, replicated in subsequent studies","pmids":["10640699"],"is_preprint":false},{"year":2002,"finding":"Cripto functions as both a coreceptor (in cis) and a secreted coligand (in trans) for Nodal signaling. The ability of Cripto to bind Nodal and mediate Nodal signaling requires addition of an O-linked fucose monosaccharide to a conserved site within EGF-CFC proteins.","method":"Luciferase reporter assay, cell coculture assays, glycosylation inhibition studies","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal functional assays, glycosylation modification mechanistically validated","pmids":["12052855"],"is_preprint":false},{"year":2002,"finding":"Cripto-1 binds to cell surface ALK4 on mammary epithelial cells (shown by FACS and co-immunoprecipitation). In the presence of Nodal and ALK4, CR-1 phosphorylates Smad2. CR-1 activation of MAPK and Akt is independent of Nodal and ALK4, indicating CR-1 modulates distinct signaling pathways.","method":"FACS binding assay, co-immunoprecipitation, phage display library screening, Smad2 phosphorylation assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, FACS binding, and functional Smad2 phosphorylation assays in the same study","pmids":["11909953"],"is_preprint":false},{"year":2003,"finding":"Cripto-1 specifically binds Glypican-1, a membrane-associated heparan sulfate proteoglycan, and activates tyrosine kinase c-Src, triggering MAPK and Akt signaling pathways independently of Nodal and ALK4. Active Src kinase is necessary for CR-1-induced in vitro transformation and migration.","method":"Binding assays, co-immunoprecipitation, kinase activity assays, transformation and migration assays with Src inhibition","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple binding and functional assays with pathway inhibition, single lab","pmids":["12649175"],"is_preprint":false},{"year":2003,"finding":"Nodal/Cripto/Alk4 pathway controls cardiomyogenesis in embryonic stem cells. Cripto signaling activates the Smad2 pathway; overexpression of activated ActRIB compensates for loss of Cripto in promoting cardiomyogenesis. Nodal antagonists inhibit Cripto-regulated cardiomyocyte induction.","method":"ES cell differentiation assay, genetic epistasis (ActRIB overexpression rescue), Nodal antagonist treatment, Smad2 activation assay","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with rescue experiment plus functional Smad2 pathway validation in ES cells","pmids":["14581455"],"is_preprint":false},{"year":2006,"finding":"Cripto binds TGF-β and reduces the association of TGF-β with its type I receptor TβRI, thereby suppressing TGF-β signaling and diminishing the cytostatic effects of TGF-β in mammary epithelial cells. siRNA knockdown of endogenous Cripto enhanced TGF-β signaling.","method":"Direct binding assay, receptor assembly co-immunoprecipitation, TGF-β reporter assay, siRNA knockdown, cytostasis assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — direct binding, receptor assembly disruption, functional reporter, and loss-of-function all in one study","pmids":["17030617"],"is_preprint":false},{"year":2007,"finding":"GPI attachment of Cripto-1 is required for its paracrine (trans) activity as a Nodal co-receptor. Soluble COOH-terminal-truncated forms of CR-1 have significantly lower activity than GPI-anchored CR-1 for inducing Nodal signaling in trans and in cis. PI-PLC treatment removes cell-surface CR-1 and renders cells refractory to Nodal.","method":"GPI omega-site identification, truncation mutant functional assays, PI-PLC treatment, Nodal signaling reporter assays, fluorescent cell-labeling paracrine assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — structure-function mutagenesis with functional validation and chemical ablation, single lab multiple methods","pmids":["17925387"],"is_preprint":false},{"year":2007,"finding":"CR-1 shedding from the membrane to a soluble form is mediated by GPI-phospholipase D (GPI-PLD) activity at the GPI-anchorage site. Growth factors (EGF, LPA, serum) trigger shedding. Soluble CR-1 promotes endothelial cell migration as a paracrine factor, while membrane-bound CR-1 stimulates endothelial sprouting through direct cell-cell contact.","method":"siRNA knockdown of GPI-PLD, GPI-PLD overexpression, conditioned medium migration assays, co-culture sprouting assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown and overexpression with functional readouts, two orthogonal approaches in one study","pmids":["17720976"],"is_preprint":false},{"year":2008,"finding":"Cripto binds the proprotein convertases Furin and PACE4 and localizes Nodal processing at the cell surface. Cripto and uncleaved Nodal associate during secretion; Cripto guides the Nodal precursor in detergent-resistant membranes to endocytic microdomains marked by GFP-Flotillin, coupling Nodal processing and endocytosis.","method":"Co-immunoprecipitation, brefeldin A treatment, density gradient fractionation, antibody uptake experiments, fluorescence imaging","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Moderate — co-IP of Furin/PACE4, fractionation, trafficking assays with multiple orthogonal methods in one study","pmids":["18772886"],"is_preprint":false},{"year":2009,"finding":"Cell surface GRP78 is a necessary mediator of Cripto signaling. Targeted disruption of the Cripto/GRP78 complex using shRNAs or GRP78 immunoneutralization blocks Cripto activation of MAPK/PI3K pathways and modulation of activin-A, activin-B, Nodal and TGF-β1 signaling, and prevents Cripto-mediated proliferation, E-cadherin downregulation, and decreased cell adhesion.","method":"shRNA knockdown, GRP78 immunoneutralization, phospho-signaling assays, proliferation and adhesion assays","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — two independent disruption approaches (shRNA and antibody neutralization) with multiple functional readouts, single lab","pmids":["19421146"],"is_preprint":false},{"year":2009,"finding":"Cripto-1 binds all four mammalian Notch receptors (confirmed by co-immunoprecipitation). The CFC domain of Cripto-1 and the C-terminal EGF-like repeats of Notch1 mediate the interaction, occurring mainly in the ER-Golgi network. Cripto-1 recruits Notch1 into lipid raft microdomains and enhances furin-like protease-mediated S1 cleavage (proteolytic maturation) of Notch1, sensitizing it to ligand-induced Notch signaling activation.","method":"Yeast two-hybrid screen, co-immunoprecipitation, deletion analysis, lipid raft fractionation, S1 cleavage assay, knockdown of Cripto-1","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — yeast two-hybrid plus co-IP plus fractionation plus functional cleavage assay plus KD, multiple orthogonal methods in one study","pmids":["19948478"],"is_preprint":false},{"year":1999,"finding":"Cripto-1 induces phosphorylation of AKT and GSK-3β via a PI3K-dependent pathway in human cervical carcinoma cells. PI3K inhibition blocks Cripto-1-mediated AKT and GSK-3β phosphorylation and leads to apoptosis, which can be partially rescued by exogenous CR-1.","method":"Phosphorylation assays, PI3K inhibitor (LY294002) treatment, apoptosis assay, rescue experiment","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — functional phosphorylation assay with inhibitor and rescue, single lab single cell line","pmids":["10493495"],"is_preprint":false},{"year":1999,"finding":"Cripto-1 indirectly induces tyrosine phosphorylation of ErbB-4 but not ErbB-2 or ErbB-3. Down-regulation of ErbB-4 impairs CR-1-mediated MAPK activation. Chemical cross-linking of 125I-CR-1 identifies two specific binding bands at 130 kDa and 60 kDa distinct from known ErbB receptors.","method":"Tyrosine phosphorylation assay, anti-ErbB-4 blocking antibody, hammerhead ribozyme, chemical crosslinking of radiolabeled CR-1","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — crosslinking and functional loss-of-function for ErbB-4, but novel receptor not identified; single lab","pmids":["10085099"],"is_preprint":false},{"year":1994,"finding":"CR-1 is a glycoprotein: N-glycosidase F digestion shifts the 27–29 kDa human CR-1 protein to 20 kDa, and tunicamycin treatment produces a similar shift. CR-1 growth stimulatory activity is not mediated through the EGF receptor, as anti-EGF receptor antibody did not inhibit CR-1 action and CR-1 failed to inhibit 125I-EGF binding.","method":"Western blot, immunoprecipitation, N-glycosidase F digestion, tunicamycin treatment, 125I-EGF radioreceptor assay, EGF receptor blocking antibody, proliferation assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — biochemical characterization of glycoprotein with negative EGF-receptor result confirmed by multiple methods, single lab","pmids":["8006041"],"is_preprint":false},{"year":2006,"finding":"Germ cell nuclear factor (GCNF) directly represses CRIPTO-1 transcription by binding to a DR0 element in the CRIPTO-1 promoter during differentiation. GCNF binding was confirmed in vitro (EMSA) and in vivo (ChIP); GCNF-VP16 fusion prevented retinoic acid-induced downregulation of CRIPTO-1.","method":"EMSA, chromatin immunoprecipitation, luciferase reporter assay, GCNF overexpression and GCNF-VP16 fusion expression","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — EMSA and ChIP identifying a DR0 element plus reporter assay and functional overexpression, multiple orthogonal methods","pmids":["16954206"],"is_preprint":false},{"year":2007,"finding":"β-Catenin/TCF/LEF directly regulate expression of the short form of human Cripto-1 through an intronic-exonic enhancer element containing three tandem TCF/LEF binding sites within the CR-1 gene.","method":"Luciferase reporter assay, TCF/LEF binding site identification and mutation, chromatin analysis in colon carcinoma and hepatoma cell lines","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — reporter assay with identified binding sites, single lab","pmids":["17291450"],"is_preprint":false},{"year":2009,"finding":"HIF-1α directly regulates Cripto-1 expression by binding to hypoxia-responsive elements in the Cripto-1 promoter. Cripto-1 is required for hypoxia to induce cardiomyocyte differentiation from mouse ES cells; hypoxia fails to induce cardiomyogenesis in Cripto-1-deficient mES cells.","method":"ChIP for HIF-1α at Cripto-1 promoter, ES cell differentiation assay under hypoxia, Cripto-1 knockout/knockdown in mES cells","journal":"The American journal of pathology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — ChIP confirming direct HIF-1α binding plus loss-of-function functional differentiation assay, multiple orthogonal methods","pmids":["19834060"],"is_preprint":false},{"year":2012,"finding":"Cripto-1 binds to Wnt co-receptors LRP5 and LRP6, facilitating Wnt3a binding to LRP5/6 and enhancing Wnt3a signaling through cytoplasmic β-catenin stabilization and elevated β-catenin/Tcf transcriptional activation.","method":"Co-immunoprecipitation, β-catenin cytoplasmic stabilization assay, TCF/β-catenin reporter assay, invasion and colony formation assays","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP and reporter assay, single lab, functional consequences shown","pmids":["23022962"],"is_preprint":false},{"year":2012,"finding":"Conditional inactivation of Cripto in adult satellite cells compromises skeletal muscle regeneration, while gain of function accelerates regeneration and leads to muscle hypertrophy. Cripto promotes myogenic cell proliferation and determination by antagonizing the TGF-β ligand myostatin.","method":"Conditional knockout in satellite cells, gain-of-function overexpression, muscle regeneration assays, myostatin antagonism functional assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — conditional KO and gain-of-function in vivo with defined phenotypic readouts, mechanistic antagonism of myostatin shown","pmids":["23129614"],"is_preprint":false},{"year":2009,"finding":"The CFC domain of Cripto interacts with the ALK4 receptor extracellular domain with a KD in the micromolar range. Residues H120 and W124 of the CFC domain, previously identified by mutagenesis, are positioned to interact at the CFC-ALK4 interface. The CFC domain does not interact with ActRIIB.","method":"NMR structural characterization of synthetic CFC domain, SPR binding studies, molecular docking","journal":"Journal of peptide science","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — NMR structure of isolated domain with SPR binding, not full protein and single lab","pmids":["19035567"],"is_preprint":false},{"year":2016,"finding":"MEF2C transcription factor directly activates Tdgf1 (Cripto) transcription in the anterior second heart field via an AHF-restricted Tdgf1 enhancer. Loss of Mef2c in the AHF causes outflow tract alignment defects, placing TDGF1 downstream of MEF2C in cardiac development.","method":"Conditional Mef2c knockout in AHF, Tdgf1 enhancer reporter assay, ChIP for MEF2C binding, genetic epistasis","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo conditional KO plus ChIP and enhancer reporter assay confirming direct transcriptional regulation","pmids":["26811383"],"is_preprint":false},{"year":2018,"finding":"Cripto-1 binds Frizzled-7 (FZD7), LRP6, and Dishevelled-3 (DVL3) of the Wnt/β-catenin pathway and stabilizes DVL3 protein, activating the Wnt/β-catenin signaling cascade to confer stemness in hepatocellular carcinoma. Forced expression of constitutively active β-catenin partially rescues stemness suppression upon Cripto-1 knockdown.","method":"Co-immunoprecipitation of FZD7/LRP6/DVL3, DVL3 protein stability assay, β-catenin reporter assay, Cripto-1 knockdown, constitutively active β-catenin rescue","journal":"Cell death and differentiation","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP with multiple Wnt pathway components and rescue experiment, single lab","pmids":["29445127"],"is_preprint":false},{"year":2016,"finding":"Cripto sustains mouse ESC self-renewal by modulating Wnt/β-catenin signaling, while it maintains mouse EpiSC and human ESC pluripotency through Nodal/Smad2 signaling. Cripto deficiency permits ESC transdifferentiation into trophectoderm lineage and attenuates ESC lineage restriction, demonstrating pathway-specific roles in different pluripotency states.","method":"Cripto knockout ES cells, Wnt/β-catenin reporter assay, Smad2 phosphorylation assay, lineage differentiation assays in vitro and in vivo, trophectoderm marker analysis","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with multiple functional differentiation readouts distinguishing Wnt vs Nodal/Smad2 pathways in different cell states","pmids":["27586544"],"is_preprint":false},{"year":2008,"finding":"Cripto-1 interacts with caveolin-1 (Cav-1) in COS7 cells and mammary epithelial cells. Cav-1 coexpression with Cr-1 markedly reduces Cr-1-mediated migration, invasion, branching, and inhibits Cr-1-mediated activation of c-Src and MAPK. In Cav-1 null/MMTV-CR-1 transgenic mammary cells, enhanced motility and MAPK/c-Src activation are observed.","method":"Co-immunoprecipitation, overexpression and co-expression studies, Cav-1 knockout mouse model (Cav-1 null/MMTV-CR-1), migration and invasion assays, signaling assays","journal":"The American journal of pathology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus genetic in vivo validation, single lab","pmids":["18202186"],"is_preprint":false},{"year":2008,"finding":"Cripto-1 blocks TGF-β1 receptor binding, phosphorylation of Smad2 and Smad3, TGF-β-responsive reporter activity, and TGF-β1-mediated senescence of keratinocytes, indicating Cripto-1 inhibits TGF-β1 signaling in skin epithelial cells.","method":"TGF-β receptor binding blockade assay, Smad2/3 phosphorylation assay, luciferase reporter assay, senescence assay, Cripto-1 treatment of primary keratinocytes","journal":"Molecular cancer research : MCR","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — multiple functional endpoints for TGF-β inhibition, single lab and cell type","pmids":["18337457"],"is_preprint":false},{"year":2013,"finding":"Conditional inactivation of Cripto during mouse gastrulation leads to accumulation of mesenchymal cells around the primitive streak and defects in definitive endoderm formation and allocation. Both the Fgf8-Fgfr1 pathway and p38 MAP kinase activation are partially affected by loss of Cripto function.","method":"Conditional Cripto knockout mouse, embryo phenotypic analysis, Fgf8-Fgfr1 pathway and p38 signaling assays","journal":"Developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — in vivo conditional KO with pathway analysis, single lab","pmids":["23747598"],"is_preprint":false},{"year":2017,"finding":"Computational modeling and molecular dynamics simulations demonstrate that Cripto binding to GRP78 completely changes GRP78 membrane dynamics, preventing GRP78 from dissociating from the membrane and thereby enabling GRP78 tumorigenic functions at the cell surface.","method":"Molecular dynamics simulation, structural modeling","journal":"Protein science","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational prediction only, no experimental validation","pmids":["29226519"],"is_preprint":false},{"year":2020,"finding":"Conditional deletion of Cripto in the myeloid lineage (CriptoMy-LOF) impairs macrophage plasticity in injured muscle and mdx dystrophic muscle: CriptoMy-LOF macrophages fail to expand as anti-inflammatory CD206+ macrophages due to aberrant TGFβ/Smad signaling activation. This reduction in plasticity disturbs vascular remodeling by increasing Endothelial-to-Mesenchymal Transition (EndMT) and exacerbates dystrophic phenotype.","method":"Cre-loxP conditional knockout in myeloid lineage, macrophage FACS phenotyping, TGFβ/Smad signaling assay, EndMT analysis in mdx mouse model","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo conditional KO with defined cellular and signaling phenotypes, multiple disease models","pmids":["32107853"],"is_preprint":false},{"year":2021,"finding":"Cripto participates in a TGF-β–ALK1–Cripto receptor complex at the plasma membrane of chondrocytes, inducing catabolic SMAD1/5 signaling and favoring hypertrophic differentiation, contributing to osteoarthritis. Elevated Cripto expression was validated in murine and human OA cartilage.","method":"Co-immunoprecipitation of TGF-β/ALK1/Cripto complex, SMAD1/5 phosphorylation assay, in vitro and ex vivo chondrocyte hypertrophy assays, OA mouse model gene expression analysis","journal":"The Journal of pathology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP of receptor complex and functional signaling assay with ex vivo validation, single lab","pmids":["34357595"],"is_preprint":false},{"year":2012,"finding":"Cripto-1 enhances JAK2/STAT3 phosphorylation in mesenchymal stem cells through a GRP78-dependent mechanism. Anti-GRP78 antibody blocks Cripto-mediated JAK2/STAT3 activation, c-Myc and cyclin D1 upregulation, and the pro-survival effects of Cripto against hypoxia/H2O2-induced apoptosis.","method":"Phosphorylation assays (p-JAK2, p-STAT3), anti-GRP78 antibody blocking, STAT3 siRNA, apoptosis assay","journal":"Annals of the New York Academy of Sciences","confidence":"Low","confidence_rationale":"Tier 3 / Weak — indirect signaling assays and antibody blocking, single lab, limited mechanistic depth in abstract","pmids":["22901256"],"is_preprint":false},{"year":2016,"finding":"Cripto-1 modulates macrophage cytokine secretion (IL-10, TNF-α, IL-6, IL-1β) and phagocytic activity via activation of NF-κB signaling, specifically through IκB kinase phosphorylation and p65 nuclear translocation. NF-κB inhibition blocks these Cripto-1-induced effects.","method":"NF-κB inhibitor (APDC) treatment, IKK phosphorylation assay, p65 nuclear translocation assay, cytokine ELISA, phagocytosis assay","journal":"Immunologic research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — multiple functional assays with inhibitor and signaling validation, single lab","pmids":["26476731"],"is_preprint":false}],"current_model":"CRIPTO (CR-1/TDGF1) is a GPI-anchored cell-surface glycoprotein that functions as an obligate co-receptor for TGF-β family ligands Nodal, GDF1, and GDF3 by forming a complex with ALK4 (via its CFC domain) and ActRIIB to activate Smad2/3 signaling; it also recruits Furin/PACE4 to direct Nodal proteolytic processing and endocytic trafficking. Independently of Nodal/ALK4, CRIPTO activates Src/MAPK/PI3K-Akt pathways via Glypican-1 and cell-surface GRP78, while directly binding and inhibiting TGF-β1 by blocking TβRI assembly. CRIPTO can be shed as a soluble paracrine factor through GPI-PLD activity, enhances Notch receptor S1 maturation by recruiting Notch into lipid rafts, promotes Wnt/β-catenin signaling by binding LRP5/6 and stabilizing Dishevelled-3, and is transcriptionally regulated by HIF-1α (under hypoxia) and repressed by GCNF (during differentiation), with Wnt/β-catenin/TCF also driving a short-form isoform."},"narrative":{"mechanistic_narrative":"CRIPTO (CR-1/TDGF1) is a GPI-anchored cell-surface glycoprotein [PMID:10640699, PMID:8006041] that acts as an obligate co-receptor for Nodal-class TGF-β ligands and, independently, as a multifunctional modulator of growth-factor signaling controlling embryonic development, tissue regeneration, and tumorigenesis [PMID:11389842, PMID:27586544]. As a Nodal co-receptor, CRIPTO engages the type I receptor ALK4 through its CFC motif—an interaction structurally mapped to residues H120/W124 and required both for Nodal binding to the ALK4/ActRIIB complex and for downstream Smad2 activation [PMID:11389842, PMID:19035567]; its co-receptor and secreted co-ligand functions depend on O-linked fucosylation of the EGF-CFC region and on intact GPI anchorage for cis and trans activity [PMID:12052855, PMID:17925387]. CRIPTO further organizes Nodal biogenesis by binding the proprotein convertases Furin and PACE4, coupling Nodal processing to endocytic trafficking in detergent-resistant membranes [PMID:18772886], and can itself be shed as a soluble paracrine factor through GPI-PLD activity [PMID:17720976]. Beyond Nodal, CRIPTO drives Src/MAPK/PI3K-Akt signaling via Glypican-1 and cell-surface GRP78 [PMID:12649175, PMID:19421146, PMID:10493495], directly binds TGF-β1 to block TβRI assembly and suppress cytostatic/senescence responses [PMID:17030617, PMID:18337457], promotes Notch1 S1 maturation by recruiting it into lipid rafts [PMID:19948478], and potentiates Wnt/β-catenin signaling by binding LRP5/6 and stabilizing Dishevelled-3 [PMID:23022962, PMID:29445127]. Through these activities CRIPTO sustains stem-cell self-renewal and pluripotency in a pathway-selective manner [PMID:27586544], governs gastrulation and cardiac development [PMID:14581455, PMID:26811383, PMID:23747598], and controls satellite-cell-driven muscle regeneration and macrophage plasticity by antagonizing TGF-β/myostatin signaling [PMID:23129614, PMID:32107853]. CRIPTO transcription is directly activated by HIF-1α under hypoxia and MEF2C in the heart field, and repressed by GCNF during differentiation, with β-catenin/TCF driving a short isoform [PMID:16954206, PMID:17291450, PMID:19834060, PMID:26811383].","teleology":[{"year":1994,"claim":"Established that CR-1 is a secreted/cell-associated glycoprotein acting through a route distinct from the EGF receptor, ruling out the obvious candidate and framing the search for its true mechanism.","evidence":"Glycosidase/tunicamycin shifts and EGF radioreceptor competition in proliferation assays","pmids":["8006041"],"confidence":"Medium","gaps":["No receptor identified","Signaling pathway undefined"]},{"year":1999,"claim":"Showed CRIPTO drives PI3K-Akt/GSK-3β survival signaling and engages ErbB-4 indirectly, hinting at receptor-independent oncogenic outputs distinct from the later-defined Nodal axis.","evidence":"Phospho-assays with PI3K inhibitor and rescue; crosslinking and ErbB-4 blockade in carcinoma cells","pmids":["10493495","10085099"],"confidence":"Medium","gaps":["The 130/60 kDa crosslinked receptor not identified","Single cell line; relation to GRP78/Glypican-1 axis unresolved at the time"]},{"year":2000,"claim":"Defined CRIPTO as GPI-anchored, establishing the membrane topology that would prove essential for its spatial co-receptor function.","evidence":"Biochemical fractionation, PI-PLC, immunofluorescence","pmids":["10640699"],"confidence":"High","gaps":["Functional consequence of GPI anchorage not yet tested","Shedding mechanism unknown"]},{"year":2001,"claim":"Identified the CFC-domain interaction with ALK4 as the molecular basis for Cripto-dependent Nodal signaling, defining its obligate co-receptor role.","evidence":"Co-IP, receptor binding and Smad2 reporter assays in cell culture","pmids":["11389842"],"confidence":"High","gaps":["Stoichiometry and structure of the ternary complex unknown","Whether ALK4 binding alone is sufficient unresolved"]},{"year":2002,"claim":"Distinguished CRIPTO's cis co-receptor from trans co-ligand activity and showed both require O-fucosylation, revealing post-translational control of ligand-receptor assembly.","evidence":"Coculture and reporter assays, glycosylation inhibition; FACS/co-IP with ALK4 and Smad2 phosphorylation","pmids":["12052855","11909953"],"confidence":"High","gaps":["MAPK/Akt arm shown Nodal/ALK4-independent but mediator not yet identified","In vivo relevance of fucosylation untested"]},{"year":2003,"claim":"Resolved the Nodal-independent arm by identifying Glypican-1 as a CRIPTO partner driving c-Src/MAPK/Akt and transformation, and placed the Nodal/Cripto/ALK4 axis in cardiomyogenic differentiation.","evidence":"Binding/co-IP, Src kinase and transformation/migration assays; ES cell differentiation with ActRIB rescue and Nodal antagonists","pmids":["12649175","14581455"],"confidence":"High","gaps":["How Glypican-1 couples to Src not mechanistically detailed","Link between the two signaling arms unclear"]},{"year":2006,"claim":"Revealed a ligand-trap function: CRIPTO binds TGF-β1 and blocks TβRI assembly, antagonizing cytostatic TGF-β signaling and offering an oncogenic mechanism independent of Nodal.","evidence":"Direct binding, receptor assembly co-IP, reporter and cytostasis assays with siRNA","pmids":["17030617"],"confidence":"High","gaps":["Structural basis of TGF-β1 binding undefined","Selectivity among TGF-β isoforms not fully mapped"]},{"year":2007,"claim":"Demonstrated that GPI anchorage is functionally required for paracrine and autocrine Nodal co-receptor activity and that GPI-PLD-mediated shedding produces a distinct soluble paracrine factor.","evidence":"Omega-site/truncation mutagenesis, PI-PLC ablation, paracrine reporter assays; GPI-PLD knockdown/overexpression with migration and sprouting assays","pmids":["17925387","17720976"],"confidence":"High","gaps":["Physiological triggers of shedding in vivo unknown","Relative contribution of membrane vs soluble CRIPTO context-dependent"]},{"year":2008,"claim":"Connected CRIPTO to Nodal biogenesis machinery by showing it recruits Furin/PACE4 to localize Nodal processing and endocytosis, and identified caveolin-1 as a negative regulator of CRIPTO oncogenic signaling.","evidence":"Co-IP, BFA, density gradient fractionation and trafficking imaging; Cav-1 co-IP and Cav-1-null/MMTV-CR-1 mouse signaling assays","pmids":["18772886","18202186"],"confidence":"High","gaps":["How CRIPTO selects Nodal for raft microdomains unclear","Cav-1 regulatory mechanism (sequestration vs signaling block) not resolved"]},{"year":2008,"claim":"Extended the TGF-β1-inhibitory mechanism to skin epithelium, showing CRIPTO blocks TGF-β1-induced Smad2/3 signaling and senescence.","evidence":"Receptor binding blockade, Smad2/3 phosphorylation, reporter and senescence assays in keratinocytes","pmids":["18337457"],"confidence":"Medium","gaps":["Single cell type","Whether membrane-bound or soluble CRIPTO mediates the effect not separated"]},{"year":2009,"claim":"Identified cell-surface GRP78 as a required mediator of CRIPTO's MAPK/PI3K signaling and modulation of multiple TGF-β ligands, and mapped CRIPTO-Notch1 interaction promoting S1 maturation, broadening its signaling reach.","evidence":"shRNA/GRP78 immunoneutralization with functional readouts; Y2H, co-IP, raft fractionation and S1 cleavage assays for Notch1","pmids":["19421146","19948478"],"confidence":"High","gaps":["GRP78-to-kinase coupling mechanism undefined","Whether Notch enhancement operates in vivo untested"]},{"year":2009,"claim":"Placed CRIPTO under direct hypoxic transcriptional control by HIF-1α and showed this drives hypoxia-induced cardiomyogenesis, linking the gene's expression to the microenvironment.","evidence":"ChIP for HIF-1α at the promoter and ES cell differentiation under hypoxia with loss-of-function","pmids":["19834060"],"confidence":"High","gaps":["Other hypoxia-responsive cofactors not examined","Relevance to tumor hypoxia not directly tested here"]},{"year":2009,"claim":"Provided structural and affinity detail of the CFC-ALK4 interface and showed CFC does not bind ActRIIB, refining the receptor-engagement model.","evidence":"NMR of synthetic CFC domain, SPR binding, molecular docking","pmids":["19035567"],"confidence":"Medium","gaps":["Isolated domain, not full-length protein","Ternary complex structure with Nodal absent"]},{"year":2006,"claim":"Defined transcriptional repression of CRIPTO by GCNF during differentiation and β-catenin/TCF-driven induction of a short isoform, establishing how CRIPTO levels track developmental state.","evidence":"EMSA, ChIP, reporter and GCNF-VP16 fusion assays; TCF/LEF site mutation reporter assays","pmids":["16954206","17291450"],"confidence":"Medium","gaps":["Function of the short isoform not characterized","Interplay between repressive and activating inputs unresolved"]},{"year":2012,"claim":"Established CRIPTO as a positive Wnt/β-catenin modulator by binding LRP5/6 and facilitating Wnt3a signaling, adding a third major signaling axis.","evidence":"Co-IP, β-catenin stabilization and TCF reporter, invasion/colony assays","pmids":["23022962"],"confidence":"Medium","gaps":["Mechanism of LRP enhancement unclear","Single lab"]},{"year":2012,"claim":"Defined an in vivo regenerative role: CRIPTO in satellite cells promotes myogenic proliferation/determination by antagonizing myostatin, demonstrating physiological TGF-β ligand-trap function.","evidence":"Conditional KO and gain-of-function in satellite cells, muscle regeneration assays, myostatin antagonism","pmids":["23129614"],"confidence":"High","gaps":["Direct myostatin-CRIPTO binding not shown here","Receptor context for antagonism undefined"]},{"year":2013,"claim":"Showed CRIPTO is required during gastrulation for endoderm formation and primitive streak organization, partly through Fgf8-Fgfr1 and p38 pathways.","evidence":"Conditional Cripto KO mouse embryo phenotyping and pathway analysis","pmids":["23747598"],"confidence":"Medium","gaps":["Whether Fgf/p38 effects are direct or downstream of Nodal unclear","Single lab"]},{"year":2016,"claim":"Demonstrated pathway-selective control of pluripotency—CRIPTO uses Wnt/β-catenin in mouse ESCs versus Nodal/Smad2 in EpiSCs/human ESCs—and placed Tdgf1 downstream of MEF2C in cardiac outflow tract development.","evidence":"Cripto KO ESCs with Wnt and Smad2 readouts and lineage assays; conditional Mef2c KO with Tdgf1 enhancer reporter and ChIP","pmids":["27586544","26811383"],"confidence":"High","gaps":["How cell state selects which pathway CRIPTO engages is unresolved","Direct enhancer dissection of stage-specific outputs incomplete"]},{"year":2016,"claim":"Implicated CRIPTO in innate immune signaling by showing it modulates macrophage cytokine output and phagocytosis through NF-κB activation.","evidence":"NF-κB inhibitor, IKK phosphorylation, p65 translocation, cytokine ELISA, phagocytosis assays","pmids":["26476731"],"confidence":"Medium","gaps":["Upstream receptor coupling to NF-κB undefined","Single lab"]},{"year":2018,"claim":"Extended the Wnt axis in cancer, showing CRIPTO binds FZD7/LRP6/DVL3 and stabilizes DVL3 to confer stemness in hepatocellular carcinoma.","evidence":"Co-IP, DVL3 stability and β-catenin reporter assays, knockdown with constitutively active β-catenin rescue","pmids":["29445127"],"confidence":"Medium","gaps":["Mechanism of DVL3 stabilization undefined","Single lab"]},{"year":2020,"claim":"Revealed a myeloid-lineage role: CRIPTO restrains TGFβ/Smad in macrophages to enable anti-inflammatory plasticity, with loss exacerbating dystrophic muscle pathology via EndMT.","evidence":"Myeloid conditional KO, macrophage FACS, TGFβ/Smad assays, EndMT analysis in mdx mice","pmids":["32107853"],"confidence":"High","gaps":["Whether CRIPTO acts cell-autonomously in macrophages vs paracrine unclear","Receptor mediating TGFβ restraint not pinned down"]},{"year":2021,"claim":"Identified a TGF-β–ALK1–Cripto complex driving catabolic SMAD1/5 signaling and chondrocyte hypertrophy in osteoarthritis, expanding CRIPTO's receptor partnerships beyond ALK4.","evidence":"Co-IP of the receptor complex, SMAD1/5 phosphorylation, chondrocyte hypertrophy assays, OA mouse/human expression","pmids":["34357595"],"confidence":"Medium","gaps":["Direct CFC-ALK1 contact not mapped","Single lab"]},{"year":null,"claim":"How CRIPTO integrates and switches between its multiple signaling axes (Nodal/Smad2, Glypican-1/GRP78/Src, TGF-β1 inhibition, Notch, Wnt) in a cell-state- and context-dependent manner remains unresolved at the structural and systems level.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No full-length structure of CRIPTO in any receptor complex","Quantitative rules governing pathway selection unknown","How membrane vs soluble CRIPTO differentially routes signaling is unresolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[0,2,5]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[6,19,25]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,4,11,18]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[9,22]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,3,9,11]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[7,8]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,4,6,11,18]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[5,21,23,26]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[9,11]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[28,31]}],"complexes":["Nodal/Cripto/ALK4/ActRIIB co-receptor complex","TGF-β–ALK1–Cripto complex"],"partners":["ALK4","GPC1","HSPA5","FURIN","PACE4","NOTCH1","LRP6","DVL3"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P13385","full_name":"Protein Cripto","aliases":["Cripto, EGF-CFC family member","Cripto-1 growth factor","CRGF","Epidermal growth factor-like cripto protein CR1","Teratocarcinoma-derived growth factor 1"],"length_aa":188,"mass_kda":21.2,"function":"GPI-anchored cell membrane protein involved in Nodal signaling. Cell-associated CRIPTO acts as a Nodal coreceptor in cis. Shedding of CRIPTO by TMEM8A modulates Nodal signaling by allowing soluble CRIPTO to act as a Nodal coreceptor on other cells (PubMed:27881714). Could play a role in the determination of the epiblastic cells that subsequently give rise to the mesoderm (PubMed:11909953)","subcellular_location":"Cell membrane; Secreted","url":"https://www.uniprot.org/uniprotkb/P13385/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CRIPTO","classification":"Common Essential","n_dependent_lines":780,"n_total_lines":1208,"dependency_fraction":0.6456953642384106},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CRIPTO","total_profiled":1310},"omim":[{"mim_id":"619342","title":"POST-GPI ATTACHMENT TO PROTEINS 6; PGAP6","url":"https://www.omim.org/entry/619342"},{"mim_id":"608981","title":"ACTIVIN A RECEPTOR, TYPE IC; ACVR1C","url":"https://www.omim.org/entry/608981"},{"mim_id":"607491","title":"PROTEIN O-FUCOSYLTRANSFERASE 1; POFUT1","url":"https://www.omim.org/entry/607491"},{"mim_id":"606522","title":"GROWTH/DIFFERENTIATION FACTOR 3; GDF3","url":"https://www.omim.org/entry/606522"},{"mim_id":"605194","title":"CRYPTIC, EGF-CFC FAMILY MEMBER 1; CFC1","url":"https://www.omim.org/entry/605194"}],"hpa":{"profiled":true,"resolved_as":"TDGF1","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"kidney","ntpm":18.8},{"tissue":"lymphoid tissue","ntpm":19.7},{"tissue":"ovary","ntpm":9.4}],"url":"https://www.proteinatlas.org/search/TDGF1"},"hgnc":{"alias_symbol":["CR","Cripto-1","CR-1"],"prev_symbol":["TDGF1"]},"alphafold":{"accession":"P13385","domains":[{"cath_id":"2.10.25.10","chopping":"75-112","consensus_level":"medium","plddt":89.1624,"start":75,"end":112},{"cath_id":"-","chopping":"115-149","consensus_level":"medium","plddt":92.0117,"start":115,"end":149}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P13385","model_url":"https://alphafold.ebi.ac.uk/files/AF-P13385-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P13385-F1-predicted_aligned_error_v6.png","plddt_mean":68.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CRIPTO","jax_strain_url":"https://www.jax.org/strain/search?query=CRIPTO"},"sequence":{"accession":"P13385","fasta_url":"https://rest.uniprot.org/uniprotkb/P13385.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P13385/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P13385"}},"corpus_meta":[{"pmid":"11389842","id":"PMC_11389842","title":"Nodal signals to Smads through Cripto-dependent and Cripto-independent mechanisms.","date":"2001","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/11389842","citation_count":317,"is_preprint":false},{"pmid":"12052855","id":"PMC_12052855","title":"Dual roles of Cripto as a ligand and coreceptor in the nodal signaling pathway.","date":"2002","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12052855","citation_count":175,"is_preprint":false},{"pmid":"19421146","id":"PMC_19421146","title":"Blockade of Cripto binding to cell surface GRP78 inhibits oncogenic Cripto signaling via MAPK/PI3K and Smad2/3 pathways.","date":"2009","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/19421146","citation_count":153,"is_preprint":false},{"pmid":"14581455","id":"PMC_14581455","title":"Nodal-dependent Cripto signaling promotes cardiomyogenesis and redirects the neural fate of embryonic stem cells.","date":"2003","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/14581455","citation_count":137,"is_preprint":false},{"pmid":"16123806","id":"PMC_16123806","title":"Cripto-1: a multifunctional modulator during embryogenesis and oncogenesis.","date":"2005","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/16123806","citation_count":136,"is_preprint":false},{"pmid":"11909953","id":"PMC_11909953","title":"Cripto-1 activates nodal- and ALK4-dependent and -independent signaling pathways in mammary epithelial Cells.","date":"2002","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/11909953","citation_count":123,"is_preprint":false},{"pmid":"8180021","id":"PMC_8180021","title":"Expression of transforming growth factor alpha, amphiregulin and cripto-1 in human breast carcinomas.","date":"1994","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/8180021","citation_count":118,"is_preprint":false},{"pmid":"7916676","id":"PMC_7916676","title":"The murine cripto gene: expression during mesoderm induction and early heart morphogenesis.","date":"1993","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/7916676","citation_count":112,"is_preprint":false},{"pmid":"10640699","id":"PMC_10640699","title":"Membrane-anchorage of Cripto protein by glycosylphosphatidylinositol and its distribution during early mouse development.","date":"2000","source":"Mechanisms of development","url":"https://pubmed.ncbi.nlm.nih.gov/10640699","citation_count":100,"is_preprint":false},{"pmid":"20616345","id":"PMC_20616345","title":"Role of Cripto-1 in stem cell maintenance and malignant progression.","date":"2010","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/20616345","citation_count":96,"is_preprint":false},{"pmid":"12649175","id":"PMC_12649175","title":"A Nodal- and ALK4-independent signaling pathway activated by Cripto-1 through Glypican-1 and c-Src.","date":"2003","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/12649175","citation_count":93,"is_preprint":false},{"pmid":"25153355","id":"PMC_25153355","title":"The multifaceted role of the embryonic gene Cripto-1 in cancer, stem cells and epithelial-mesenchymal transition.","date":"2014","source":"Seminars in cancer biology","url":"https://pubmed.ncbi.nlm.nih.gov/25153355","citation_count":92,"is_preprint":false},{"pmid":"22542493","id":"PMC_22542493","title":"Role of Cripto-1 during epithelial-to-mesenchymal transition in development and cancer.","date":"2012","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/22542493","citation_count":89,"is_preprint":false},{"pmid":"8006041","id":"PMC_8006041","title":"Identification and biological characterization of an epidermal growth factor-related protein: cripto-1.","date":"1994","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8006041","citation_count":89,"is_preprint":false},{"pmid":"12073012","id":"PMC_12073012","title":"A loss-of-function mutation in the CFC domain of TDGF1 is associated with human forebrain defects.","date":"2002","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/12073012","citation_count":85,"is_preprint":false},{"pmid":"17030617","id":"PMC_17030617","title":"Cripto binds transforming growth factor beta (TGF-beta) and inhibits TGF-beta signaling.","date":"2006","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/17030617","citation_count":80,"is_preprint":false},{"pmid":"11339828","id":"PMC_11339828","title":"Cripto-1 enhances migration and branching morphogenesis of mouse mammary epithelial cells.","date":"2001","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/11339828","citation_count":77,"is_preprint":false},{"pmid":"8588926","id":"PMC_8588926","title":"Detection and location of amphiregulin and Cripto-1 expression in the developing postnatal mouse mammary gland.","date":"1995","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/8588926","citation_count":74,"is_preprint":false},{"pmid":"7579500","id":"PMC_7579500","title":"Expression of messenger RNA for amphiregulin, heregulin, and cripto-1, three new members of the epidermal growth factor family, in human breast carcinomas.","date":"1995","source":"Breast cancer research and treatment","url":"https://pubmed.ncbi.nlm.nih.gov/7579500","citation_count":74,"is_preprint":false},{"pmid":"22306319","id":"PMC_22306319","title":"Cripto/GRP78 modulation of the TGF-β pathway in development and oncogenesis.","date":"2012","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/22306319","citation_count":73,"is_preprint":false},{"pmid":"16951234","id":"PMC_16951234","title":"Identification of cripto-1 as a novel serologic marker for breast and colon cancer.","date":"2006","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/16951234","citation_count":71,"is_preprint":false},{"pmid":"21573645","id":"PMC_21573645","title":"Expression of amphiregulin, cripto-1, and heregulin-alpha in human breast-cancer cells.","date":"1993","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/21573645","citation_count":70,"is_preprint":false},{"pmid":"18772886","id":"PMC_18772886","title":"Cripto recruits Furin and PACE4 and controls Nodal trafficking during proteolytic maturation.","date":"2008","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/18772886","citation_count":69,"is_preprint":false},{"pmid":"15657343","id":"PMC_15657343","title":"Role of human cripto-1 in tumor angiogenesis.","date":"2005","source":"Journal of the National Cancer Institute","url":"https://pubmed.ncbi.nlm.nih.gov/15657343","citation_count":67,"is_preprint":false},{"pmid":"10070255","id":"PMC_10070255","title":"Cripto: a novel epidermal growth factor (EGF)-related peptide in mammary gland development and neoplasia.","date":"1999","source":"BioEssays : news and reviews in molecular, cellular and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/10070255","citation_count":65,"is_preprint":false},{"pmid":"10085099","id":"PMC_10085099","title":"Cripto-1 indirectly stimulates the tyrosine phosphorylation of erb B-4 through a novel receptor.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10085099","citation_count":65,"is_preprint":false},{"pmid":"12370739","id":"PMC_12370739","title":"Transforming growth factor alpha, amphiregulin and cripto-1 are frequently expressed in advanced human ovarian carcinomas.","date":"2002","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/12370739","citation_count":65,"is_preprint":false},{"pmid":"15949532","id":"PMC_15949532","title":"Cripto-1: an oncofetal gene with many faces.","date":"2005","source":"Current topics in developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/15949532","citation_count":64,"is_preprint":false},{"pmid":"29445127","id":"PMC_29445127","title":"Cripto-1 contributes to stemness in hepatocellular carcinoma by stabilizing Dishevelled-3 and activating Wnt/β-catenin pathway.","date":"2018","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/29445127","citation_count":64,"is_preprint":false},{"pmid":"17720976","id":"PMC_17720976","title":"Growth factor induction of Cripto-1 shedding by glycosylphosphatidylinositol-phospholipase D and enhancement of endothelial cell migration.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17720976","citation_count":60,"is_preprint":false},{"pmid":"11857442","id":"PMC_11857442","title":"Cripto: a tumor growth factor and more.","date":"2002","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/11857442","citation_count":59,"is_preprint":false},{"pmid":"26059540","id":"PMC_26059540","title":"Cripto-1 as a novel therapeutic target for triple negative breast cancer.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26059540","citation_count":58,"is_preprint":false},{"pmid":"14584041","id":"PMC_14584041","title":"Cripto-1 overexpression leads to enhanced invasiveness and resistance to anoikis in human MCF-7 breast cancer cells.","date":"2004","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/14584041","citation_count":58,"is_preprint":false},{"pmid":"15173016","id":"PMC_15173016","title":"Cripto: a novel target for antibody-based cancer immunotherapy.","date":"2004","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/15173016","citation_count":54,"is_preprint":false},{"pmid":"10493495","id":"PMC_10493495","title":"Cripto-1 induces phosphatidylinositol 3'-kinase-dependent phosphorylation of AKT and glycogen synthase kinase 3beta in human cervical carcinoma cells.","date":"1999","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/10493495","citation_count":54,"is_preprint":false},{"pmid":"19029628","id":"PMC_19029628","title":"Emerging roles of nodal and Cripto-1: from embryogenesis to breast cancer progression.","date":"2008","source":"Breast disease","url":"https://pubmed.ncbi.nlm.nih.gov/19029628","citation_count":53,"is_preprint":false},{"pmid":"20549704","id":"PMC_20549704","title":"Cripto-1 is a cell surface marker for a tumorigenic, undifferentiated subpopulation in human embryonal carcinoma cells.","date":"2010","source":"Stem cells (Dayton, Ohio)","url":"https://pubmed.ncbi.nlm.nih.gov/20549704","citation_count":51,"is_preprint":false},{"pmid":"27586544","id":"PMC_27586544","title":"Cripto is essential to capture mouse epiblast stem cell and human embryonic stem cell pluripotency.","date":"2016","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/27586544","citation_count":51,"is_preprint":false},{"pmid":"16123800","id":"PMC_16123800","title":"Nodal-dependant Cripto signaling in ES cells: from stem cells to tumor biology.","date":"2005","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/16123800","citation_count":50,"is_preprint":false},{"pmid":"22149969","id":"PMC_22149969","title":"An evolving web of signaling networks regulated by Cripto-1.","date":"2011","source":"Growth factors (Chur, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/22149969","citation_count":50,"is_preprint":false},{"pmid":"26343543","id":"PMC_26343543","title":"Dynamic regulation of the cancer stem cell compartment by Cripto-1 in colorectal cancer.","date":"2015","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/26343543","citation_count":50,"is_preprint":false},{"pmid":"17925387","id":"PMC_17925387","title":"Requirement of glycosylphosphatidylinositol anchor of Cripto-1 for trans activity as a Nodal co-receptor.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17925387","citation_count":50,"is_preprint":false},{"pmid":"20624125","id":"PMC_20624125","title":"Cripto-1: an embryonic gene that promotes tumorigenesis.","date":"2010","source":"Future oncology (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/20624125","citation_count":49,"is_preprint":false},{"pmid":"26811383","id":"PMC_26811383","title":"MEF2C regulates outflow tract alignment and transcriptional control of Tdgf1.","date":"2016","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/26811383","citation_count":49,"is_preprint":false},{"pmid":"19834060","id":"PMC_19834060","title":"Cripto-1 is required for hypoxia to induce cardiac differentiation of mouse embryonic stem cells.","date":"2009","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/19834060","citation_count":46,"is_preprint":false},{"pmid":"19948478","id":"PMC_19948478","title":"Enhancement of Notch receptor maturation and signaling sensitivity by Cripto-1.","date":"2009","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/19948478","citation_count":44,"is_preprint":false},{"pmid":"8561865","id":"PMC_8561865","title":"Detection of amphiregulin and Cripto-1 in mammary tumors from transgenic mice.","date":"1996","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/8561865","citation_count":44,"is_preprint":false},{"pmid":"23022962","id":"PMC_23022962","title":"Cripto-1 enhances the canonical Wnt/β-catenin signaling pathway by binding to LRP5 and LRP6 co-receptors.","date":"2012","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/23022962","citation_count":43,"is_preprint":false},{"pmid":"10200494","id":"PMC_10200494","title":"Cripto: roles in mammary cell growth, survival, differentiation and transformation.","date":"1998","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/10200494","citation_count":42,"is_preprint":false},{"pmid":"21862554","id":"PMC_21862554","title":"Regulation of extra-embryonic endoderm stem cell differentiation by Nodal and Cripto signaling.","date":"2011","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/21862554","citation_count":42,"is_preprint":false},{"pmid":"18604175","id":"PMC_18604175","title":"Potential for cripto-1 in defining stem cell-like characteristics in human malignant melanoma.","date":"2008","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/18604175","citation_count":41,"is_preprint":false},{"pmid":"23129614","id":"PMC_23129614","title":"Cripto regulates skeletal muscle regeneration and modulates satellite cell determination by antagonizing myostatin.","date":"2012","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/23129614","citation_count":40,"is_preprint":false},{"pmid":"11992720","id":"PMC_11992720","title":"Role of the EGF-CFC gene cripto in cell differentiation and embryo development.","date":"2002","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/11992720","citation_count":37,"is_preprint":false},{"pmid":"9570042","id":"PMC_9570042","title":"Purification and characterization of a recombinant human cripto-1 protein.","date":"1998","source":"Growth factors (Chur, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/9570042","citation_count":34,"is_preprint":false},{"pmid":"28394345","id":"PMC_28394345","title":"CRIPTO and its signaling partner GRP78 drive the metastatic phenotype in human osteotropic prostate cancer.","date":"2017","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/28394345","citation_count":34,"is_preprint":false},{"pmid":"16954206","id":"PMC_16954206","title":"Germ cell nuclear factor is a repressor of CRIPTO-1 and CRIPTO-3.","date":"2006","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16954206","citation_count":33,"is_preprint":false},{"pmid":"15300009","id":"PMC_15300009","title":"Nodal and Cripto-1: embryonic pattern formation genes involved in mammary gland development and tumorigenesis.","date":"2004","source":"Journal of mammary gland biology and neoplasia","url":"https://pubmed.ncbi.nlm.nih.gov/15300009","citation_count":31,"is_preprint":false},{"pmid":"21824804","id":"PMC_21824804","title":"The role of Cripto-1 in the tumorigenesis and progression of oral squamous cell carcinoma.","date":"2011","source":"Oral oncology","url":"https://pubmed.ncbi.nlm.nih.gov/21824804","citation_count":31,"is_preprint":false},{"pmid":"21863025","id":"PMC_21863025","title":"Expression and functional role of CRIPTO-1 in cutaneous melanoma.","date":"2011","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/21863025","citation_count":30,"is_preprint":false},{"pmid":"30143536","id":"PMC_30143536","title":"Cripto-1 Plasmid DNA Vaccination Targets Metastasis and Cancer Stem Cells in Murine Mammary Carcinoma.","date":"2018","source":"Cancer immunology research","url":"https://pubmed.ncbi.nlm.nih.gov/30143536","citation_count":30,"is_preprint":false},{"pmid":"24521322","id":"PMC_24521322","title":"Cripto-1 expression in glioblastoma multiforme.","date":"2014","source":"Brain pathology (Zurich, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/24521322","citation_count":29,"is_preprint":false},{"pmid":"21073352","id":"PMC_21073352","title":"Targeting the embryonic gene Cripto-1 in cancer and beyond.","date":"2010","source":"Expert opinion on therapeutic patents","url":"https://pubmed.ncbi.nlm.nih.gov/21073352","citation_count":29,"is_preprint":false},{"pmid":"17291450","id":"PMC_17291450","title":"beta-Catenin/TCF/LEF regulate expression of the short form human Cripto-1.","date":"2007","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/17291450","citation_count":29,"is_preprint":false},{"pmid":"15737733","id":"PMC_15737733","title":"Context-dependent neuronal differentiation and germ layer induction of Smad4-/- and Cripto-/- embryonic stem cells.","date":"2005","source":"Molecular and cellular neurosciences","url":"https://pubmed.ncbi.nlm.nih.gov/15737733","citation_count":29,"is_preprint":false},{"pmid":"35295489","id":"PMC_35295489","title":"Fractalkine/CX3CR1 Pathway in Neuropathic Pain: An Update.","date":"2021","source":"Frontiers in pain research (Lausanne, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/35295489","citation_count":28,"is_preprint":false},{"pmid":"19302412","id":"PMC_19302412","title":"Riding shotgun: a dual role for the epidermal growth factor-Cripto/FRL-1/Cryptic protein Cripto in Nodal trafficking.","date":"2009","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/19302412","citation_count":28,"is_preprint":false},{"pmid":"29604056","id":"PMC_29604056","title":"CRIPTO promotes an aggressive tumour phenotype and resistance to treatment in hepatocellular carcinoma.","date":"2018","source":"The Journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/29604056","citation_count":28,"is_preprint":false},{"pmid":"19732464","id":"PMC_19732464","title":"Cripto-1 overexpression is involved in the tumorigenesis of nasopharyngeal carcinoma.","date":"2009","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/19732464","citation_count":26,"is_preprint":false},{"pmid":"22901256","id":"PMC_22901256","title":"Hematopoietic stem cells are regulated by Cripto, as an intermediary of HIF-1α in the hypoxic bone marrow niche.","date":"2012","source":"Annals of the New York Academy of Sciences","url":"https://pubmed.ncbi.nlm.nih.gov/22901256","citation_count":26,"is_preprint":false},{"pmid":"8635871","id":"PMC_8635871","title":"Transfection with a CRIPTO anti-sense plasmid suppresses endogenous CRIPTO expression and inhibits transformation in a human embryonal carcinoma cell line.","date":"1996","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/8635871","citation_count":26,"is_preprint":false},{"pmid":"28835002","id":"PMC_28835002","title":"Cripto Enhances Proliferation and Survival of Mesenchymal Stem Cells by Up-Regulating JAK2/STAT3 Pathway in a GRP78-Dependent Manner.","date":"2018","source":"Biomolecules & therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/28835002","citation_count":25,"is_preprint":false},{"pmid":"26654129","id":"PMC_26654129","title":"Cripto: Expression, epigenetic regulation and potential diagnostic use in testicular germ cell tumors.","date":"2015","source":"Molecular oncology","url":"https://pubmed.ncbi.nlm.nih.gov/26654129","citation_count":24,"is_preprint":false},{"pmid":"8661720","id":"PMC_8661720","title":"Characterization of the mouse Tdgf1 gene and Tdgf pseudogenes.","date":"1996","source":"Mammalian genome : official journal of the International Mammalian Genome Society","url":"https://pubmed.ncbi.nlm.nih.gov/8661720","citation_count":23,"is_preprint":false},{"pmid":"26476731","id":"PMC_26476731","title":"Cripto-1 modulates macrophage cytokine secretion and phagocytic activity via NF-κB signaling.","date":"2016","source":"Immunologic research","url":"https://pubmed.ncbi.nlm.nih.gov/26476731","citation_count":22,"is_preprint":false},{"pmid":"28098905","id":"PMC_28098905","title":"Cripto-1 promotes epithelial-mesenchymal transition in prostate cancer via Wnt/β-catenin signaling.","date":"2017","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/28098905","citation_count":22,"is_preprint":false},{"pmid":"23574716","id":"PMC_23574716","title":"The significance of a Cripto-1 positive subpopulation of human melanoma cells exhibiting stem cell-like characteristics.","date":"2013","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/23574716","citation_count":22,"is_preprint":false},{"pmid":"18337457","id":"PMC_18337457","title":"Cripto-1 alters keratinocyte differentiation via blockade of transforming growth factor-beta1 signaling: role in skin carcinogenesis.","date":"2008","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/18337457","citation_count":22,"is_preprint":false},{"pmid":"31455359","id":"PMC_31455359","title":"Cripto-1 expression in patients with clear cell renal cell carcinoma is associated with poor disease outcome.","date":"2019","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/31455359","citation_count":21,"is_preprint":false},{"pmid":"27467944","id":"PMC_27467944","title":"Cripto-1 vaccination elicits protective immunity against metastatic melanoma.","date":"2016","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/27467944","citation_count":21,"is_preprint":false},{"pmid":"23784832","id":"PMC_23784832","title":"Nodal/Cripto signaling in fetal male germ cell development: implications for testicular germ cell tumors.","date":"2013","source":"The International journal of developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/23784832","citation_count":20,"is_preprint":false},{"pmid":"32107853","id":"PMC_32107853","title":"Cripto shapes macrophage plasticity and restricts EndMT in injured and diseased skeletal muscle.","date":"2020","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/32107853","citation_count":20,"is_preprint":false},{"pmid":"20128024","id":"PMC_20128024","title":"Significance and relationship between Cripto-1 and p-STAT3 expression in gastric cancer and precancerous lesions.","date":"2010","source":"World journal of gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/20128024","citation_count":20,"is_preprint":false},{"pmid":"20126975","id":"PMC_20126975","title":"TDGF1 is a novel predictive marker for metachronous metastasis of colorectal cancer.","date":"2010","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/20126975","citation_count":19,"is_preprint":false},{"pmid":"11807813","id":"PMC_11807813","title":"Detection and localization of Cripto-1 binding in mouse mammary epithelial cells and in the mouse mammary gland using an immunoglobulin-cripto-1 fusion protein.","date":"2002","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/11807813","citation_count":19,"is_preprint":false},{"pmid":"18202186","id":"PMC_18202186","title":"Regulation of Cripto-1 signaling and biological activity by caveolin-1 in mammary epithelial cells.","date":"2008","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/18202186","citation_count":19,"is_preprint":false},{"pmid":"19853938","id":"PMC_19853938","title":"Teratocarcinoma-derived growth factor 1 (TDGF1) sequence variants in patients with congenital heart defect.","date":"2009","source":"International journal of cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/19853938","citation_count":18,"is_preprint":false},{"pmid":"36790376","id":"PMC_36790376","title":"CX3CR1+ Macrophage Facilitates the Resolution of Allergic Lung Inflammation via Interacting CCL26.","date":"2023","source":"American journal of respiratory and critical care medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36790376","citation_count":17,"is_preprint":false},{"pmid":"21496809","id":"PMC_21496809","title":"Altered expression of activin, cripto, and follistatin in the endometrium of women with endometrioma.","date":"2011","source":"Fertility and sterility","url":"https://pubmed.ncbi.nlm.nih.gov/21496809","citation_count":17,"is_preprint":false},{"pmid":"30703478","id":"PMC_30703478","title":"Development of conformational antibodies targeting Cripto-1 with neutralizing effects in vitro.","date":"2019","source":"Biochimie","url":"https://pubmed.ncbi.nlm.nih.gov/30703478","citation_count":17,"is_preprint":false},{"pmid":"34502188","id":"PMC_34502188","title":"Cripto-1 as a Key Factor in Tumor Progression, Epithelial to Mesenchymal Transition and Cancer Stem Cells.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34502188","citation_count":16,"is_preprint":false},{"pmid":"10024674","id":"PMC_10024674","title":"Expression of cripto and amphiregulin in colon mucosa from high risk colon cancer families.","date":"1999","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/10024674","citation_count":16,"is_preprint":false},{"pmid":"23747598","id":"PMC_23747598","title":"Cripto is required for mesoderm and endoderm cell allocation during mouse gastrulation.","date":"2013","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/23747598","citation_count":15,"is_preprint":false},{"pmid":"17412323","id":"PMC_17412323","title":"Cripto-1 expression in uveal melanoma: an immunohistochemical study.","date":"2007","source":"Experimental eye research","url":"https://pubmed.ncbi.nlm.nih.gov/17412323","citation_count":14,"is_preprint":false},{"pmid":"34357595","id":"PMC_34357595","title":"Cripto favors chondrocyte hypertrophy via TGF-β SMAD1/5 signaling during development of osteoarthritis.","date":"2021","source":"The Journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/34357595","citation_count":14,"is_preprint":false},{"pmid":"25228630","id":"PMC_25228630","title":"Expression of Nodal, Cripto, SMAD3, phosphorylated SMAD3, and SMAD4 in the proliferative endometrium of women with endometriosis.","date":"2014","source":"Reproductive sciences (Thousand Oaks, Calif.)","url":"https://pubmed.ncbi.nlm.nih.gov/25228630","citation_count":14,"is_preprint":false},{"pmid":"19035567","id":"PMC_19035567","title":"Structural insights into the interaction between the Cripto CFC domain and the ALK4 receptor.","date":"2009","source":"Journal of peptide science : an official publication of the European Peptide Society","url":"https://pubmed.ncbi.nlm.nih.gov/19035567","citation_count":14,"is_preprint":false},{"pmid":"22182448","id":"PMC_22182448","title":"Human recombinant Cripto-1 increases doubling time and reduces proliferation of HeLa cells independent of pro-proliferation pathways.","date":"2011","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/22182448","citation_count":14,"is_preprint":false},{"pmid":"34065315","id":"PMC_34065315","title":"Cripto-1 as a Potential Target of Cancer Stem Cells for Immunotherapy.","date":"2021","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/34065315","citation_count":13,"is_preprint":false},{"pmid":"29108289","id":"PMC_29108289","title":"Clinical significance of cripto-1 expression in lung adenocarcinoma.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29108289","citation_count":13,"is_preprint":false},{"pmid":"29226519","id":"PMC_29226519","title":"Cripto stabilizes GRP78 on the cell membrane.","date":"2017","source":"Protein science : a publication of the Protein Society","url":"https://pubmed.ncbi.nlm.nih.gov/29226519","citation_count":13,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":47737,"output_tokens":7899,"usd":0.130848,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":17417,"output_tokens":6662,"usd":0.126817,"stage2_stop_reason":"end_turn"},"total_usd":0.257665,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2001,\n      \"finding\": \"Cripto interacts with the type I receptor ALK4 via its conserved CFC motif. This interaction is necessary both for Nodal binding to the ALK4/ActRIIB receptor complex and for Smad2 activation by Nodal.\",\n      \"method\": \"Co-immunoprecipitation, receptor binding assays, Smad2 activation reporter assays in cell culture\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding and functional activation assays, replicated across multiple labs in subsequent papers\",\n      \"pmids\": [\"11389842\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Cripto protein is anchored to the cell membrane via a glycosylphosphatidylinositol (GPI) moiety, as demonstrated by biochemical characterization and immunofluorescence; GPI-linkage is proposed to be important for spatial specificity of cell-cell interactions.\",\n      \"method\": \"Biochemical fractionation, PI-PLC treatment, immunofluorescence\",\n      \"journal\": \"Mechanisms of development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal biochemical methods, replicated in subsequent studies\",\n      \"pmids\": [\"10640699\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Cripto functions as both a coreceptor (in cis) and a secreted coligand (in trans) for Nodal signaling. The ability of Cripto to bind Nodal and mediate Nodal signaling requires addition of an O-linked fucose monosaccharide to a conserved site within EGF-CFC proteins.\",\n      \"method\": \"Luciferase reporter assay, cell coculture assays, glycosylation inhibition studies\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal functional assays, glycosylation modification mechanistically validated\",\n      \"pmids\": [\"12052855\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Cripto-1 binds to cell surface ALK4 on mammary epithelial cells (shown by FACS and co-immunoprecipitation). In the presence of Nodal and ALK4, CR-1 phosphorylates Smad2. CR-1 activation of MAPK and Akt is independent of Nodal and ALK4, indicating CR-1 modulates distinct signaling pathways.\",\n      \"method\": \"FACS binding assay, co-immunoprecipitation, phage display library screening, Smad2 phosphorylation assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, FACS binding, and functional Smad2 phosphorylation assays in the same study\",\n      \"pmids\": [\"11909953\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Cripto-1 specifically binds Glypican-1, a membrane-associated heparan sulfate proteoglycan, and activates tyrosine kinase c-Src, triggering MAPK and Akt signaling pathways independently of Nodal and ALK4. Active Src kinase is necessary for CR-1-induced in vitro transformation and migration.\",\n      \"method\": \"Binding assays, co-immunoprecipitation, kinase activity assays, transformation and migration assays with Src inhibition\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple binding and functional assays with pathway inhibition, single lab\",\n      \"pmids\": [\"12649175\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Nodal/Cripto/Alk4 pathway controls cardiomyogenesis in embryonic stem cells. Cripto signaling activates the Smad2 pathway; overexpression of activated ActRIB compensates for loss of Cripto in promoting cardiomyogenesis. Nodal antagonists inhibit Cripto-regulated cardiomyocyte induction.\",\n      \"method\": \"ES cell differentiation assay, genetic epistasis (ActRIB overexpression rescue), Nodal antagonist treatment, Smad2 activation assay\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with rescue experiment plus functional Smad2 pathway validation in ES cells\",\n      \"pmids\": [\"14581455\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Cripto binds TGF-β and reduces the association of TGF-β with its type I receptor TβRI, thereby suppressing TGF-β signaling and diminishing the cytostatic effects of TGF-β in mammary epithelial cells. siRNA knockdown of endogenous Cripto enhanced TGF-β signaling.\",\n      \"method\": \"Direct binding assay, receptor assembly co-immunoprecipitation, TGF-β reporter assay, siRNA knockdown, cytostasis assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding, receptor assembly disruption, functional reporter, and loss-of-function all in one study\",\n      \"pmids\": [\"17030617\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"GPI attachment of Cripto-1 is required for its paracrine (trans) activity as a Nodal co-receptor. Soluble COOH-terminal-truncated forms of CR-1 have significantly lower activity than GPI-anchored CR-1 for inducing Nodal signaling in trans and in cis. PI-PLC treatment removes cell-surface CR-1 and renders cells refractory to Nodal.\",\n      \"method\": \"GPI omega-site identification, truncation mutant functional assays, PI-PLC treatment, Nodal signaling reporter assays, fluorescent cell-labeling paracrine assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — structure-function mutagenesis with functional validation and chemical ablation, single lab multiple methods\",\n      \"pmids\": [\"17925387\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"CR-1 shedding from the membrane to a soluble form is mediated by GPI-phospholipase D (GPI-PLD) activity at the GPI-anchorage site. Growth factors (EGF, LPA, serum) trigger shedding. Soluble CR-1 promotes endothelial cell migration as a paracrine factor, while membrane-bound CR-1 stimulates endothelial sprouting through direct cell-cell contact.\",\n      \"method\": \"siRNA knockdown of GPI-PLD, GPI-PLD overexpression, conditioned medium migration assays, co-culture sprouting assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown and overexpression with functional readouts, two orthogonal approaches in one study\",\n      \"pmids\": [\"17720976\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Cripto binds the proprotein convertases Furin and PACE4 and localizes Nodal processing at the cell surface. Cripto and uncleaved Nodal associate during secretion; Cripto guides the Nodal precursor in detergent-resistant membranes to endocytic microdomains marked by GFP-Flotillin, coupling Nodal processing and endocytosis.\",\n      \"method\": \"Co-immunoprecipitation, brefeldin A treatment, density gradient fractionation, antibody uptake experiments, fluorescence imaging\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP of Furin/PACE4, fractionation, trafficking assays with multiple orthogonal methods in one study\",\n      \"pmids\": [\"18772886\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Cell surface GRP78 is a necessary mediator of Cripto signaling. Targeted disruption of the Cripto/GRP78 complex using shRNAs or GRP78 immunoneutralization blocks Cripto activation of MAPK/PI3K pathways and modulation of activin-A, activin-B, Nodal and TGF-β1 signaling, and prevents Cripto-mediated proliferation, E-cadherin downregulation, and decreased cell adhesion.\",\n      \"method\": \"shRNA knockdown, GRP78 immunoneutralization, phospho-signaling assays, proliferation and adhesion assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — two independent disruption approaches (shRNA and antibody neutralization) with multiple functional readouts, single lab\",\n      \"pmids\": [\"19421146\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Cripto-1 binds all four mammalian Notch receptors (confirmed by co-immunoprecipitation). The CFC domain of Cripto-1 and the C-terminal EGF-like repeats of Notch1 mediate the interaction, occurring mainly in the ER-Golgi network. Cripto-1 recruits Notch1 into lipid raft microdomains and enhances furin-like protease-mediated S1 cleavage (proteolytic maturation) of Notch1, sensitizing it to ligand-induced Notch signaling activation.\",\n      \"method\": \"Yeast two-hybrid screen, co-immunoprecipitation, deletion analysis, lipid raft fractionation, S1 cleavage assay, knockdown of Cripto-1\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast two-hybrid plus co-IP plus fractionation plus functional cleavage assay plus KD, multiple orthogonal methods in one study\",\n      \"pmids\": [\"19948478\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Cripto-1 induces phosphorylation of AKT and GSK-3β via a PI3K-dependent pathway in human cervical carcinoma cells. PI3K inhibition blocks Cripto-1-mediated AKT and GSK-3β phosphorylation and leads to apoptosis, which can be partially rescued by exogenous CR-1.\",\n      \"method\": \"Phosphorylation assays, PI3K inhibitor (LY294002) treatment, apoptosis assay, rescue experiment\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — functional phosphorylation assay with inhibitor and rescue, single lab single cell line\",\n      \"pmids\": [\"10493495\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Cripto-1 indirectly induces tyrosine phosphorylation of ErbB-4 but not ErbB-2 or ErbB-3. Down-regulation of ErbB-4 impairs CR-1-mediated MAPK activation. Chemical cross-linking of 125I-CR-1 identifies two specific binding bands at 130 kDa and 60 kDa distinct from known ErbB receptors.\",\n      \"method\": \"Tyrosine phosphorylation assay, anti-ErbB-4 blocking antibody, hammerhead ribozyme, chemical crosslinking of radiolabeled CR-1\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — crosslinking and functional loss-of-function for ErbB-4, but novel receptor not identified; single lab\",\n      \"pmids\": [\"10085099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"CR-1 is a glycoprotein: N-glycosidase F digestion shifts the 27–29 kDa human CR-1 protein to 20 kDa, and tunicamycin treatment produces a similar shift. CR-1 growth stimulatory activity is not mediated through the EGF receptor, as anti-EGF receptor antibody did not inhibit CR-1 action and CR-1 failed to inhibit 125I-EGF binding.\",\n      \"method\": \"Western blot, immunoprecipitation, N-glycosidase F digestion, tunicamycin treatment, 125I-EGF radioreceptor assay, EGF receptor blocking antibody, proliferation assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — biochemical characterization of glycoprotein with negative EGF-receptor result confirmed by multiple methods, single lab\",\n      \"pmids\": [\"8006041\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Germ cell nuclear factor (GCNF) directly represses CRIPTO-1 transcription by binding to a DR0 element in the CRIPTO-1 promoter during differentiation. GCNF binding was confirmed in vitro (EMSA) and in vivo (ChIP); GCNF-VP16 fusion prevented retinoic acid-induced downregulation of CRIPTO-1.\",\n      \"method\": \"EMSA, chromatin immunoprecipitation, luciferase reporter assay, GCNF overexpression and GCNF-VP16 fusion expression\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — EMSA and ChIP identifying a DR0 element plus reporter assay and functional overexpression, multiple orthogonal methods\",\n      \"pmids\": [\"16954206\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"β-Catenin/TCF/LEF directly regulate expression of the short form of human Cripto-1 through an intronic-exonic enhancer element containing three tandem TCF/LEF binding sites within the CR-1 gene.\",\n      \"method\": \"Luciferase reporter assay, TCF/LEF binding site identification and mutation, chromatin analysis in colon carcinoma and hepatoma cell lines\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — reporter assay with identified binding sites, single lab\",\n      \"pmids\": [\"17291450\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"HIF-1α directly regulates Cripto-1 expression by binding to hypoxia-responsive elements in the Cripto-1 promoter. Cripto-1 is required for hypoxia to induce cardiomyocyte differentiation from mouse ES cells; hypoxia fails to induce cardiomyogenesis in Cripto-1-deficient mES cells.\",\n      \"method\": \"ChIP for HIF-1α at Cripto-1 promoter, ES cell differentiation assay under hypoxia, Cripto-1 knockout/knockdown in mES cells\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP confirming direct HIF-1α binding plus loss-of-function functional differentiation assay, multiple orthogonal methods\",\n      \"pmids\": [\"19834060\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Cripto-1 binds to Wnt co-receptors LRP5 and LRP6, facilitating Wnt3a binding to LRP5/6 and enhancing Wnt3a signaling through cytoplasmic β-catenin stabilization and elevated β-catenin/Tcf transcriptional activation.\",\n      \"method\": \"Co-immunoprecipitation, β-catenin cytoplasmic stabilization assay, TCF/β-catenin reporter assay, invasion and colony formation assays\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP and reporter assay, single lab, functional consequences shown\",\n      \"pmids\": [\"23022962\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Conditional inactivation of Cripto in adult satellite cells compromises skeletal muscle regeneration, while gain of function accelerates regeneration and leads to muscle hypertrophy. Cripto promotes myogenic cell proliferation and determination by antagonizing the TGF-β ligand myostatin.\",\n      \"method\": \"Conditional knockout in satellite cells, gain-of-function overexpression, muscle regeneration assays, myostatin antagonism functional assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO and gain-of-function in vivo with defined phenotypic readouts, mechanistic antagonism of myostatin shown\",\n      \"pmids\": [\"23129614\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"The CFC domain of Cripto interacts with the ALK4 receptor extracellular domain with a KD in the micromolar range. Residues H120 and W124 of the CFC domain, previously identified by mutagenesis, are positioned to interact at the CFC-ALK4 interface. The CFC domain does not interact with ActRIIB.\",\n      \"method\": \"NMR structural characterization of synthetic CFC domain, SPR binding studies, molecular docking\",\n      \"journal\": \"Journal of peptide science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — NMR structure of isolated domain with SPR binding, not full protein and single lab\",\n      \"pmids\": [\"19035567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"MEF2C transcription factor directly activates Tdgf1 (Cripto) transcription in the anterior second heart field via an AHF-restricted Tdgf1 enhancer. Loss of Mef2c in the AHF causes outflow tract alignment defects, placing TDGF1 downstream of MEF2C in cardiac development.\",\n      \"method\": \"Conditional Mef2c knockout in AHF, Tdgf1 enhancer reporter assay, ChIP for MEF2C binding, genetic epistasis\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo conditional KO plus ChIP and enhancer reporter assay confirming direct transcriptional regulation\",\n      \"pmids\": [\"26811383\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Cripto-1 binds Frizzled-7 (FZD7), LRP6, and Dishevelled-3 (DVL3) of the Wnt/β-catenin pathway and stabilizes DVL3 protein, activating the Wnt/β-catenin signaling cascade to confer stemness in hepatocellular carcinoma. Forced expression of constitutively active β-catenin partially rescues stemness suppression upon Cripto-1 knockdown.\",\n      \"method\": \"Co-immunoprecipitation of FZD7/LRP6/DVL3, DVL3 protein stability assay, β-catenin reporter assay, Cripto-1 knockdown, constitutively active β-catenin rescue\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP with multiple Wnt pathway components and rescue experiment, single lab\",\n      \"pmids\": [\"29445127\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Cripto sustains mouse ESC self-renewal by modulating Wnt/β-catenin signaling, while it maintains mouse EpiSC and human ESC pluripotency through Nodal/Smad2 signaling. Cripto deficiency permits ESC transdifferentiation into trophectoderm lineage and attenuates ESC lineage restriction, demonstrating pathway-specific roles in different pluripotency states.\",\n      \"method\": \"Cripto knockout ES cells, Wnt/β-catenin reporter assay, Smad2 phosphorylation assay, lineage differentiation assays in vitro and in vivo, trophectoderm marker analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with multiple functional differentiation readouts distinguishing Wnt vs Nodal/Smad2 pathways in different cell states\",\n      \"pmids\": [\"27586544\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Cripto-1 interacts with caveolin-1 (Cav-1) in COS7 cells and mammary epithelial cells. Cav-1 coexpression with Cr-1 markedly reduces Cr-1-mediated migration, invasion, branching, and inhibits Cr-1-mediated activation of c-Src and MAPK. In Cav-1 null/MMTV-CR-1 transgenic mammary cells, enhanced motility and MAPK/c-Src activation are observed.\",\n      \"method\": \"Co-immunoprecipitation, overexpression and co-expression studies, Cav-1 knockout mouse model (Cav-1 null/MMTV-CR-1), migration and invasion assays, signaling assays\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus genetic in vivo validation, single lab\",\n      \"pmids\": [\"18202186\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Cripto-1 blocks TGF-β1 receptor binding, phosphorylation of Smad2 and Smad3, TGF-β-responsive reporter activity, and TGF-β1-mediated senescence of keratinocytes, indicating Cripto-1 inhibits TGF-β1 signaling in skin epithelial cells.\",\n      \"method\": \"TGF-β receptor binding blockade assay, Smad2/3 phosphorylation assay, luciferase reporter assay, senescence assay, Cripto-1 treatment of primary keratinocytes\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — multiple functional endpoints for TGF-β inhibition, single lab and cell type\",\n      \"pmids\": [\"18337457\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Conditional inactivation of Cripto during mouse gastrulation leads to accumulation of mesenchymal cells around the primitive streak and defects in definitive endoderm formation and allocation. Both the Fgf8-Fgfr1 pathway and p38 MAP kinase activation are partially affected by loss of Cripto function.\",\n      \"method\": \"Conditional Cripto knockout mouse, embryo phenotypic analysis, Fgf8-Fgfr1 pathway and p38 signaling assays\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — in vivo conditional KO with pathway analysis, single lab\",\n      \"pmids\": [\"23747598\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Computational modeling and molecular dynamics simulations demonstrate that Cripto binding to GRP78 completely changes GRP78 membrane dynamics, preventing GRP78 from dissociating from the membrane and thereby enabling GRP78 tumorigenic functions at the cell surface.\",\n      \"method\": \"Molecular dynamics simulation, structural modeling\",\n      \"journal\": \"Protein science\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational prediction only, no experimental validation\",\n      \"pmids\": [\"29226519\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Conditional deletion of Cripto in the myeloid lineage (CriptoMy-LOF) impairs macrophage plasticity in injured muscle and mdx dystrophic muscle: CriptoMy-LOF macrophages fail to expand as anti-inflammatory CD206+ macrophages due to aberrant TGFβ/Smad signaling activation. This reduction in plasticity disturbs vascular remodeling by increasing Endothelial-to-Mesenchymal Transition (EndMT) and exacerbates dystrophic phenotype.\",\n      \"method\": \"Cre-loxP conditional knockout in myeloid lineage, macrophage FACS phenotyping, TGFβ/Smad signaling assay, EndMT analysis in mdx mouse model\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo conditional KO with defined cellular and signaling phenotypes, multiple disease models\",\n      \"pmids\": [\"32107853\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Cripto participates in a TGF-β–ALK1–Cripto receptor complex at the plasma membrane of chondrocytes, inducing catabolic SMAD1/5 signaling and favoring hypertrophic differentiation, contributing to osteoarthritis. Elevated Cripto expression was validated in murine and human OA cartilage.\",\n      \"method\": \"Co-immunoprecipitation of TGF-β/ALK1/Cripto complex, SMAD1/5 phosphorylation assay, in vitro and ex vivo chondrocyte hypertrophy assays, OA mouse model gene expression analysis\",\n      \"journal\": \"The Journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP of receptor complex and functional signaling assay with ex vivo validation, single lab\",\n      \"pmids\": [\"34357595\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Cripto-1 enhances JAK2/STAT3 phosphorylation in mesenchymal stem cells through a GRP78-dependent mechanism. Anti-GRP78 antibody blocks Cripto-mediated JAK2/STAT3 activation, c-Myc and cyclin D1 upregulation, and the pro-survival effects of Cripto against hypoxia/H2O2-induced apoptosis.\",\n      \"method\": \"Phosphorylation assays (p-JAK2, p-STAT3), anti-GRP78 antibody blocking, STAT3 siRNA, apoptosis assay\",\n      \"journal\": \"Annals of the New York Academy of Sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — indirect signaling assays and antibody blocking, single lab, limited mechanistic depth in abstract\",\n      \"pmids\": [\"22901256\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Cripto-1 modulates macrophage cytokine secretion (IL-10, TNF-α, IL-6, IL-1β) and phagocytic activity via activation of NF-κB signaling, specifically through IκB kinase phosphorylation and p65 nuclear translocation. NF-κB inhibition blocks these Cripto-1-induced effects.\",\n      \"method\": \"NF-κB inhibitor (APDC) treatment, IKK phosphorylation assay, p65 nuclear translocation assay, cytokine ELISA, phagocytosis assay\",\n      \"journal\": \"Immunologic research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — multiple functional assays with inhibitor and signaling validation, single lab\",\n      \"pmids\": [\"26476731\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CRIPTO (CR-1/TDGF1) is a GPI-anchored cell-surface glycoprotein that functions as an obligate co-receptor for TGF-β family ligands Nodal, GDF1, and GDF3 by forming a complex with ALK4 (via its CFC domain) and ActRIIB to activate Smad2/3 signaling; it also recruits Furin/PACE4 to direct Nodal proteolytic processing and endocytic trafficking. Independently of Nodal/ALK4, CRIPTO activates Src/MAPK/PI3K-Akt pathways via Glypican-1 and cell-surface GRP78, while directly binding and inhibiting TGF-β1 by blocking TβRI assembly. CRIPTO can be shed as a soluble paracrine factor through GPI-PLD activity, enhances Notch receptor S1 maturation by recruiting Notch into lipid rafts, promotes Wnt/β-catenin signaling by binding LRP5/6 and stabilizing Dishevelled-3, and is transcriptionally regulated by HIF-1α (under hypoxia) and repressed by GCNF (during differentiation), with Wnt/β-catenin/TCF also driving a short-form isoform.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CRIPTO (CR-1/TDGF1) is a GPI-anchored cell-surface glycoprotein [#1, #14] that acts as an obligate co-receptor for Nodal-class TGF-\\u03b2 ligands and, independently, as a multifunctional modulator of growth-factor signaling controlling embryonic development, tissue regeneration, and tumorigenesis [#0, #23]. As a Nodal co-receptor, CRIPTO engages the type I receptor ALK4 through its CFC motif\\u2014an interaction structurally mapped to residues H120/W124 and required both for Nodal binding to the ALK4/ActRIIB complex and for downstream Smad2 activation [#0, #20]; its co-receptor and secreted co-ligand functions depend on O-linked fucosylation of the EGF-CFC region and on intact GPI anchorage for cis and trans activity [#2, #7]. CRIPTO further organizes Nodal biogenesis by binding the proprotein convertases Furin and PACE4, coupling Nodal processing to endocytic trafficking in detergent-resistant membranes [#9], and can itself be shed as a soluble paracrine factor through GPI-PLD activity [#8]. Beyond Nodal, CRIPTO drives Src/MAPK/PI3K-Akt signaling via Glypican-1 and cell-surface GRP78 [#4, #10, #12], directly binds TGF-\\u03b21 to block T\\u03b2RI assembly and suppress cytostatic/senescence responses [#6, #25], promotes Notch1 S1 maturation by recruiting it into lipid rafts [#11], and potentiates Wnt/\\u03b2-catenin signaling by binding LRP5/6 and stabilizing Dishevelled-3 [#18, #22]. Through these activities CRIPTO sustains stem-cell self-renewal and pluripotency in a pathway-selective manner [#23], governs gastrulation and cardiac development [#5, #21, #26], and controls satellite-cell-driven muscle regeneration and macrophage plasticity by antagonizing TGF-\\u03b2/myostatin signaling [#19, #28]. CRIPTO transcription is directly activated by HIF-1\\u03b1 under hypoxia and MEF2C in the heart field, and repressed by GCNF during differentiation, with \\u03b2-catenin/TCF driving a short isoform [#15, #16, #17, #21].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 1994,\n      \"claim\": \"Established that CR-1 is a secreted/cell-associated glycoprotein acting through a route distinct from the EGF receptor, ruling out the obvious candidate and framing the search for its true mechanism.\",\n      \"evidence\": \"Glycosidase/tunicamycin shifts and EGF radioreceptor competition in proliferation assays\",\n      \"pmids\": [\"8006041\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No receptor identified\", \"Signaling pathway undefined\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Showed CRIPTO drives PI3K-Akt/GSK-3\\u03b2 survival signaling and engages ErbB-4 indirectly, hinting at receptor-independent oncogenic outputs distinct from the later-defined Nodal axis.\",\n      \"evidence\": \"Phospho-assays with PI3K inhibitor and rescue; crosslinking and ErbB-4 blockade in carcinoma cells\",\n      \"pmids\": [\"10493495\", \"10085099\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"The 130/60 kDa crosslinked receptor not identified\", \"Single cell line; relation to GRP78/Glypican-1 axis unresolved at the time\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Defined CRIPTO as GPI-anchored, establishing the membrane topology that would prove essential for its spatial co-receptor function.\",\n      \"evidence\": \"Biochemical fractionation, PI-PLC, immunofluorescence\",\n      \"pmids\": [\"10640699\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of GPI anchorage not yet tested\", \"Shedding mechanism unknown\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Identified the CFC-domain interaction with ALK4 as the molecular basis for Cripto-dependent Nodal signaling, defining its obligate co-receptor role.\",\n      \"evidence\": \"Co-IP, receptor binding and Smad2 reporter assays in cell culture\",\n      \"pmids\": [\"11389842\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and structure of the ternary complex unknown\", \"Whether ALK4 binding alone is sufficient unresolved\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Distinguished CRIPTO's cis co-receptor from trans co-ligand activity and showed both require O-fucosylation, revealing post-translational control of ligand-receptor assembly.\",\n      \"evidence\": \"Coculture and reporter assays, glycosylation inhibition; FACS/co-IP with ALK4 and Smad2 phosphorylation\",\n      \"pmids\": [\"12052855\", \"11909953\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"MAPK/Akt arm shown Nodal/ALK4-independent but mediator not yet identified\", \"In vivo relevance of fucosylation untested\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Resolved the Nodal-independent arm by identifying Glypican-1 as a CRIPTO partner driving c-Src/MAPK/Akt and transformation, and placed the Nodal/Cripto/ALK4 axis in cardiomyogenic differentiation.\",\n      \"evidence\": \"Binding/co-IP, Src kinase and transformation/migration assays; ES cell differentiation with ActRIB rescue and Nodal antagonists\",\n      \"pmids\": [\"12649175\", \"14581455\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How Glypican-1 couples to Src not mechanistically detailed\", \"Link between the two signaling arms unclear\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Revealed a ligand-trap function: CRIPTO binds TGF-\\u03b21 and blocks T\\u03b2RI assembly, antagonizing cytostatic TGF-\\u03b2 signaling and offering an oncogenic mechanism independent of Nodal.\",\n      \"evidence\": \"Direct binding, receptor assembly co-IP, reporter and cytostasis assays with siRNA\",\n      \"pmids\": [\"17030617\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of TGF-\\u03b21 binding undefined\", \"Selectivity among TGF-\\u03b2 isoforms not fully mapped\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Demonstrated that GPI anchorage is functionally required for paracrine and autocrine Nodal co-receptor activity and that GPI-PLD-mediated shedding produces a distinct soluble paracrine factor.\",\n      \"evidence\": \"Omega-site/truncation mutagenesis, PI-PLC ablation, paracrine reporter assays; GPI-PLD knockdown/overexpression with migration and sprouting assays\",\n      \"pmids\": [\"17925387\", \"17720976\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological triggers of shedding in vivo unknown\", \"Relative contribution of membrane vs soluble CRIPTO context-dependent\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Connected CRIPTO to Nodal biogenesis machinery by showing it recruits Furin/PACE4 to localize Nodal processing and endocytosis, and identified caveolin-1 as a negative regulator of CRIPTO oncogenic signaling.\",\n      \"evidence\": \"Co-IP, BFA, density gradient fractionation and trafficking imaging; Cav-1 co-IP and Cav-1-null/MMTV-CR-1 mouse signaling assays\",\n      \"pmids\": [\"18772886\", \"18202186\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How CRIPTO selects Nodal for raft microdomains unclear\", \"Cav-1 regulatory mechanism (sequestration vs signaling block) not resolved\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Extended the TGF-\\u03b21-inhibitory mechanism to skin epithelium, showing CRIPTO blocks TGF-\\u03b21-induced Smad2/3 signaling and senescence.\",\n      \"evidence\": \"Receptor binding blockade, Smad2/3 phosphorylation, reporter and senescence assays in keratinocytes\",\n      \"pmids\": [\"18337457\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single cell type\", \"Whether membrane-bound or soluble CRIPTO mediates the effect not separated\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Identified cell-surface GRP78 as a required mediator of CRIPTO's MAPK/PI3K signaling and modulation of multiple TGF-\\u03b2 ligands, and mapped CRIPTO-Notch1 interaction promoting S1 maturation, broadening its signaling reach.\",\n      \"evidence\": \"shRNA/GRP78 immunoneutralization with functional readouts; Y2H, co-IP, raft fractionation and S1 cleavage assays for Notch1\",\n      \"pmids\": [\"19421146\", \"19948478\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"GRP78-to-kinase coupling mechanism undefined\", \"Whether Notch enhancement operates in vivo untested\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Placed CRIPTO under direct hypoxic transcriptional control by HIF-1\\u03b1 and showed this drives hypoxia-induced cardiomyogenesis, linking the gene's expression to the microenvironment.\",\n      \"evidence\": \"ChIP for HIF-1\\u03b1 at the promoter and ES cell differentiation under hypoxia with loss-of-function\",\n      \"pmids\": [\"19834060\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Other hypoxia-responsive cofactors not examined\", \"Relevance to tumor hypoxia not directly tested here\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Provided structural and affinity detail of the CFC-ALK4 interface and showed CFC does not bind ActRIIB, refining the receptor-engagement model.\",\n      \"evidence\": \"NMR of synthetic CFC domain, SPR binding, molecular docking\",\n      \"pmids\": [\"19035567\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Isolated domain, not full-length protein\", \"Ternary complex structure with Nodal absent\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Defined transcriptional repression of CRIPTO by GCNF during differentiation and \\u03b2-catenin/TCF-driven induction of a short isoform, establishing how CRIPTO levels track developmental state.\",\n      \"evidence\": \"EMSA, ChIP, reporter and GCNF-VP16 fusion assays; TCF/LEF site mutation reporter assays\",\n      \"pmids\": [\"16954206\", \"17291450\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Function of the short isoform not characterized\", \"Interplay between repressive and activating inputs unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Established CRIPTO as a positive Wnt/\\u03b2-catenin modulator by binding LRP5/6 and facilitating Wnt3a signaling, adding a third major signaling axis.\",\n      \"evidence\": \"Co-IP, \\u03b2-catenin stabilization and TCF reporter, invasion/colony assays\",\n      \"pmids\": [\"23022962\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of LRP enhancement unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined an in vivo regenerative role: CRIPTO in satellite cells promotes myogenic proliferation/determination by antagonizing myostatin, demonstrating physiological TGF-\\u03b2 ligand-trap function.\",\n      \"evidence\": \"Conditional KO and gain-of-function in satellite cells, muscle regeneration assays, myostatin antagonism\",\n      \"pmids\": [\"23129614\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct myostatin-CRIPTO binding not shown here\", \"Receptor context for antagonism undefined\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Showed CRIPTO is required during gastrulation for endoderm formation and primitive streak organization, partly through Fgf8-Fgfr1 and p38 pathways.\",\n      \"evidence\": \"Conditional Cripto KO mouse embryo phenotyping and pathway analysis\",\n      \"pmids\": [\"23747598\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether Fgf/p38 effects are direct or downstream of Nodal unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrated pathway-selective control of pluripotency\\u2014CRIPTO uses Wnt/\\u03b2-catenin in mouse ESCs versus Nodal/Smad2 in EpiSCs/human ESCs\\u2014and placed Tdgf1 downstream of MEF2C in cardiac outflow tract development.\",\n      \"evidence\": \"Cripto KO ESCs with Wnt and Smad2 readouts and lineage assays; conditional Mef2c KO with Tdgf1 enhancer reporter and ChIP\",\n      \"pmids\": [\"27586544\", \"26811383\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How cell state selects which pathway CRIPTO engages is unresolved\", \"Direct enhancer dissection of stage-specific outputs incomplete\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Implicated CRIPTO in innate immune signaling by showing it modulates macrophage cytokine output and phagocytosis through NF-\\u03baB activation.\",\n      \"evidence\": \"NF-\\u03baB inhibitor, IKK phosphorylation, p65 translocation, cytokine ELISA, phagocytosis assays\",\n      \"pmids\": [\"26476731\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Upstream receptor coupling to NF-\\u03baB undefined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Extended the Wnt axis in cancer, showing CRIPTO binds FZD7/LRP6/DVL3 and stabilizes DVL3 to confer stemness in hepatocellular carcinoma.\",\n      \"evidence\": \"Co-IP, DVL3 stability and \\u03b2-catenin reporter assays, knockdown with constitutively active \\u03b2-catenin rescue\",\n      \"pmids\": [\"29445127\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of DVL3 stabilization undefined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Revealed a myeloid-lineage role: CRIPTO restrains TGF\\u03b2/Smad in macrophages to enable anti-inflammatory plasticity, with loss exacerbating dystrophic muscle pathology via EndMT.\",\n      \"evidence\": \"Myeloid conditional KO, macrophage FACS, TGF\\u03b2/Smad assays, EndMT analysis in mdx mice\",\n      \"pmids\": [\"32107853\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether CRIPTO acts cell-autonomously in macrophages vs paracrine unclear\", \"Receptor mediating TGF\\u03b2 restraint not pinned down\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified a TGF-\\u03b2\\u2013ALK1\\u2013Cripto complex driving catabolic SMAD1/5 signaling and chondrocyte hypertrophy in osteoarthritis, expanding CRIPTO's receptor partnerships beyond ALK4.\",\n      \"evidence\": \"Co-IP of the receptor complex, SMAD1/5 phosphorylation, chondrocyte hypertrophy assays, OA mouse/human expression\",\n      \"pmids\": [\"34357595\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct CFC-ALK1 contact not mapped\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How CRIPTO integrates and switches between its multiple signaling axes (Nodal/Smad2, Glypican-1/GRP78/Src, TGF-\\u03b21 inhibition, Notch, Wnt) in a cell-state- and context-dependent manner remains unresolved at the structural and systems level.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No full-length structure of CRIPTO in any receptor complex\", \"Quantitative rules governing pathway selection unknown\", \"How membrane vs soluble CRIPTO differentially routes signaling is unresolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [0, 2, 5]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [6, 19, 25]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 4, 11, 18]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [9, 22]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 3, 9, 11]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [7, 8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 4, 6, 11, 18]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [5, 21, 23, 26]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [9, 11]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [28, 31]}\n    ],\n    \"complexes\": [\n      \"Nodal/Cripto/ALK4/ActRIIB co-receptor complex\",\n      \"TGF-\\u03b2\\u2013ALK1\\u2013Cripto complex\"\n    ],\n    \"partners\": [\n      \"ALK4\",\n      \"GPC1\",\n      \"HSPA5\",\n      \"FURIN\",\n      \"PACE4\",\n      \"NOTCH1\",\n      \"LRP6\",\n      \"DVL3\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}