{"gene":"ASPN","run_date":"2026-06-09T22:02:44","timeline":{"discoveries":[{"year":2007,"finding":"PLAP-1/asporin directly binds BMP-2, as demonstrated by co-immunoprecipitation, and negatively regulates BMP-2-induced cytodifferentiation and mineralization of periodontal ligament (PDL) cells; overexpression inhibited mineralization while knockdown enhanced BMP-2-induced differentiation.","method":"Co-immunoprecipitation, overexpression, RNAi knockdown, immunohistochemistry co-localization","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP plus gain- and loss-of-function experiments in the same study, replicated in subsequent papers","pmids":["17522060"],"is_preprint":false},{"year":2008,"finding":"PLAP-1/asporin inhibits BMP-2 signaling by competitively preventing BMP-2 from binding to BMP receptor-IB (BMPR-IB), thereby blocking Smad activation; the leucine-rich repeat 5 (LRR5) motif is the functional domain mediating this interaction, as LRR5 mutation rescues inhibition and a 26-aa LRR5-derived peptide recapitulates inhibition.","method":"Recombinant protein competition binding assay, site-directed mutagenesis of LRR5, peptide inhibition assay, Smad phosphorylation western blot","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro receptor-binding competition assay combined with mutagenesis and peptide rescue in a single study","pmids":["18407830"],"is_preprint":false},{"year":2014,"finding":"The aspartic acid (D) repeat polymorphism of PLAP-1/asporin influences its functional potency: D14-PLAP-1 suppresses BMP-2-induced cytodifferentiation more strongly than D13-PLAP-1 and shows stronger binding affinity to BMP-2 by co-immunoprecipitation.","method":"Stable transfection of D13 vs D14 cell lines, alkaline phosphatase and alizarin red staining, co-immunoprecipitation, western blot, luciferase reporter assay","journal":"Journal of dental research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus functional reporter and staining assays in single lab, multiple orthogonal methods","pmids":["24453179"],"is_preprint":false},{"year":2015,"finding":"PLAP-1/asporin positively regulates FGF-2 activity by directly binding FGF-2 and promoting FGF-2–FGFR1 complex formation; Plap-1 knockout MEFs show defective FGF-2 responses rescued by Plap-1 transfection, and reduced FGF-2/FGFR1 co-localization.","method":"Plap-1 knockout mouse-derived MEFs, binding assay, immunocytochemistry co-localization, rescue transfection","journal":"Journal of dental research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with rescue, direct binding assay, and co-localization in a single study with multiple orthogonal methods","pmids":["26239644"],"is_preprint":false},{"year":2015,"finding":"PLAP-1/asporin directly binds TLR2 and TLR4 (shown by immunoprecipitation), suppresses TLR2/4-induced NF-κB activity, reduces IκBα kinase degradation, and downregulates proinflammatory cytokine expression in PDL cells and macrophages.","method":"Overexpression, recombinant protein treatment, immunoprecipitation, NF-κB luciferase reporter, western blot for IκBα, cytokine ELISA","journal":"Journal of dental research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — Co-IP plus multiple downstream functional readouts (NF-κB activity, IκBα, cytokines) in single lab with orthogonal methods","pmids":["26399972"],"is_preprint":false},{"year":2021,"finding":"PLAP-1/asporin enhances adipogenesis: recombinant PLAP-1 promotes lipid accumulation in 3T3-L1 cells, Plap-1 knockout mice and Plap-1-knockdown 3T3-L1 cells show reduced lipid accumulation, and primary preadipocytes from Plap-1 KO mice exhibit less adipogenic differentiation than wild-type.","method":"Plap-1 knockout mouse model, siRNA knockdown in 3T3-L1 cells, recombinant protein treatment, lipid staining/accumulation assays","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO plus siRNA and recombinant protein gain-of-function in single lab","pmids":["33654143"],"is_preprint":false},{"year":2019,"finding":"1,25(OH)2D3 suppresses PLAP-1 expression transcriptionally through a vitamin D response element (VDRE) in the PLAP-1 promoter that binds VDR, as confirmed by ChIP assay and reporter gene assays, leading to enhanced osteogenic differentiation of hPDLSCs under inflammatory conditions.","method":"ChIP assay, luciferase reporter assay, western blot, RT-qPCR","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus reporter gene assay identifying specific VDRE, single lab with two orthogonal methods","pmids":["31837573"],"is_preprint":false},{"year":2022,"finding":"PLAP-1/asporin suppresses HIF-1α signaling: hypoxia-induced PLAP-1 expression is HIF-1α-dependent (blocked by chetomin), and recombinant PLAP-1 or PLAP-1 gene transfection reduces hypoxia-induced HRE-luciferase activity and nuclear HIF-1α accumulation in PDL cells.","method":"Hypoxic cell culture, chemical inhibitors, luciferase HRE reporter assay, western blot for nuclear HIF-1α, recombinant protein and transfection experiments","journal":"Journal of periodontal research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — reporter assay and western blot in single lab, multiple conditions but limited orthogonal validation","pmids":["35138637"],"is_preprint":false},{"year":2022,"finding":"PLAP-1/asporin knockout mice display enlarged periodontal ligament space, increased collagen fibril diameter, altered ECM protein expression (elevated Col3, BGN, DCN), reduced tooth extraction force, and accelerated alveolar bone resorption with more osteoclasts in ligature-induced periodontitis, establishing a structural and protective role for PLAP-1 in PDL collagen organization and periodontal inflammation.","method":"PLAP-1 KO mouse model, micro-CT, histology (HE, picrosirius red), fluorescence immunostaining, TEM, tooth extraction force measurement, ligature-induced periodontitis model","journal":"International journal of molecular sciences","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with multiple orthogonal structural and functional readouts in vivo","pmids":["37958972"],"is_preprint":false},{"year":2014,"finding":"Overexpression of PLAP-1 in rat bone marrow stromal cells (rBMSCs) inhibits their differentiation into osteoblast-like cells, reducing mineralized nodule formation and decreasing expression of osteoblast markers (Runx2, Osterix, ALP, BSP, osteocalcin).","method":"Stable retroviral transfection of PLAP-1 in rBMSCs, alizarin red staining, RT-qPCR for osteogenic markers","journal":"Journal of molecular histology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — gain-of-function in cell culture with multiple marker readouts, single lab","pmids":["25038933"],"is_preprint":false},{"year":2015,"finding":"Overexpression of PLAP-1 in rBMSCs transplanted into rat critical-size skull defects inhibits new bone formation and mineralization in vivo, confirming its role as a negative regulator of osteogenesis in a skeletal repair context.","method":"In vivo rat skull defect model, rBMSC transplantation, X-ray, HE/Masson/von Kossa histology, immunohistochemistry","journal":"Journal of molecular histology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — in vivo KO/OE model with histological readouts, single lab","pmids":["26031659"],"is_preprint":false},{"year":2023,"finding":"ASPN interacts with HAPLN1, and their combined knockdown synergistically increases ALP, OPN, OCN, and COL1A1 expression and ECM mineralization in BMSCs while decreasing osteoclast markers in bone marrow macrophages, indicating ASPN and HAPLN1 cooperate to inhibit osteogenic differentiation.","method":"Co-IP/binding analysis, siRNA knockdown in BMSCs from OVX mice, western blot for osteogenic and osteoclast markers, ECM mineralization assay","journal":"Orthopaedic surgery","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — protein interaction analysis plus siRNA functional assays in single lab","pmids":["37427673"],"is_preprint":false},{"year":2022,"finding":"ASPN mediates oxaliplatin (OXA) resistance in colorectal cancer; siRNA-mediated ASPN knockdown reverses OXA resistance and promotes cell apoptosis both in vitro and in patient-derived xenograft models in vivo.","method":"siRNA knockdown, in vitro cell viability/apoptosis assays, patient-derived xenograft (PDX) mouse model, nanoparticle co-delivery system","journal":"Biomaterials","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined phenotypic readout in vitro and PDX in vivo, single lab","pmids":["36228517"],"is_preprint":false},{"year":2026,"finding":"Exosomal miR-143-5p from H. pylori-infected epithelial cells functions as a nuclear activating microRNA (NamiRNA) that binds the super-enhancer region of the ASPN gene, increases H3K27ac enrichment, and promotes ASPN transcription in fibroblasts, leading to upregulation of pro-inflammatory cytokines (IL-4, IL-6, TGF-β); in vivo antagomir-143-5p reduced ASPN and cytokine expression and alleviated gastric inflammation.","method":"MicroRNA sequencing, co-culture assay, immunofluorescence, ChIP for H3K27ac, in vivo antagomir treatment","journal":"Gut pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus co-culture, in vivo antagomir, and immunofluorescence in single lab with orthogonal methods","pmids":["41723544"],"is_preprint":false},{"year":2024,"finding":"ASPN overexpression amplifies MATN3-driven gastric cancer cell proliferation, migration, and invasion, and MATN3-ASPN protein-protein interaction was confirmed; co-overexpression of MATN3 and ASPN enhanced tumor growth and metastasis in vivo.","method":"Protein-protein interaction analysis, co-expression analysis, overexpression/knockdown functional assays (proliferation, migration, invasion, apoptosis), in vivo mouse tumor model","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — protein interaction plus in vitro and in vivo functional assays, single lab","pmids":["39301785"],"is_preprint":false}],"current_model":"ASPN/PLAP-1 is an extracellular matrix small leucine-rich repeat proteoglycan that acts as a multi-target negative regulator of growth factor signaling: it directly binds BMP-2 via its LRR5 motif to competitively block BMPR-IB engagement and downstream Smad activation, directly binds TGF-β1 to inhibit TGF-β/Smad signaling, and directly binds FGF-2 to promote FGF-2–FGFR1 complex formation (a positive regulatory role); it also binds TLR2 and TLR4 to suppress NF-κB-mediated inflammatory cytokine production, suppresses HIF-1α transcriptional activity in a feedback loop, regulates collagen fibril organization in the periodontal ligament, promotes adipogenesis, and inhibits osteogenic differentiation of PDL and bone marrow stromal cells, with the N-terminal aspartic acid repeat polymorphism modulating the strength of BMP-2 binding and inhibition."},"narrative":{"mechanistic_narrative":"ASPN (PLAP-1/asporin) is an extracellular matrix small leucine-rich repeat proteoglycan that acts as a multi-target regulator of growth factor and inflammatory signaling and as a structural organizer of connective tissue [PMID:17522060, PMID:37958972]. It directly binds BMP-2 and inhibits BMP-2-induced cytodifferentiation and mineralization, competitively blocking BMP-2 engagement of BMP receptor-IB and the downstream Smad cascade through its LRR5 motif, which alone is sufficient to recapitulate inhibition [PMID:17522060, PMID:18407830]; the N-terminal aspartic acid repeat length tunes this activity, with the D14 variant binding BMP-2 more strongly and suppressing differentiation more potently than D13 [PMID:24453179]. In contrast to its inhibitory role toward BMP-2, ASPN positively regulates FGF-2 by binding it and promoting FGF-2–FGFR1 complex formation [PMID:26239644]. ASPN also binds TLR2 and TLR4 to dampen NF-κB activation, stabilizing IκBα and reducing proinflammatory cytokine output [PMID:26399972], and it suppresses HIF-1α transcriptional activity in a hypoxia-induced feedback loop [PMID:35138637]. Consistent with a matrix-organizing function, ASPN-deficient mice show enlarged periodontal ligament space, altered collagen fibril architecture, and accelerated alveolar bone resorption [PMID:37958972]. Across mesenchymal lineages ASPN restrains osteogenic differentiation of periodontal ligament and bone marrow stromal cells while promoting adipogenesis [PMID:33654143, PMID:25038933, PMID:26031659]. In cancer, ASPN contributes to oxaliplatin resistance in colorectal cancer and amplifies MATN3-driven gastric cancer progression [PMID:36228517, PMID:39301785].","teleology":[{"year":2007,"claim":"Established ASPN as a direct binding partner and negative regulator of BMP-2 signaling, defining its first molecular function beyond being a matrix proteoglycan.","evidence":"Co-immunoprecipitation with gain- and loss-of-function in periodontal ligament cells","pmids":["17522060"],"confidence":"High","gaps":["Did not resolve the binding domain or whether inhibition was receptor-competitive","Limited to PDL cell context"]},{"year":2008,"claim":"Resolved the mechanism of BMP-2 inhibition as competitive blockade of BMPR-IB and mapped the activity to the LRR5 motif, converting a binding observation into a defined structure-function model.","evidence":"Recombinant competition binding, LRR5 mutagenesis, and a 26-aa peptide rescue with Smad phosphorylation readout","pmids":["18407830"],"confidence":"High","gaps":["No structural model of the ASPN–BMP-2 interface","Relevance in vivo not tested"]},{"year":2014,"claim":"Showed the N-terminal aspartic acid repeat polymorphism tunes ASPN function, linking an allelic variant to differential BMP-2 binding strength and inhibitory potency.","evidence":"D13 vs D14 stable cell lines, Co-IP, reporter and mineralization assays","pmids":["24453179"],"confidence":"Medium","gaps":["Single-lab Co-IP for affinity difference","Molecular basis for repeat-dependent affinity not defined"]},{"year":2014,"claim":"Extended the anti-osteogenic role beyond PDL by showing ASPN inhibits osteoblast differentiation of bone marrow stromal cells.","evidence":"Retroviral overexpression in rBMSCs with osteogenic marker and mineralization assays","pmids":["25038933"],"confidence":"Medium","gaps":["Gain-of-function only","Mechanism downstream of BMP/FGF not separated"]},{"year":2015,"claim":"Revealed a context-dependent positive regulatory role: ASPN binds FGF-2 and promotes FGF-2–FGFR1 complex formation, contrasting with its BMP-2 inhibition.","evidence":"Plap-1 knockout MEFs with rescue, binding assay, and FGF-2/FGFR1 co-localization","pmids":["26239644"],"confidence":"High","gaps":["How ASPN can be inhibitory for BMP-2 yet stimulatory for FGF-2 mechanistically unresolved","Domain mediating FGF-2 binding not mapped"]},{"year":2015,"claim":"Defined an anti-inflammatory function through direct TLR2/TLR4 binding and suppression of NF-κB signaling.","evidence":"Immunoprecipitation, NF-κB reporter, IκBα western blot, cytokine ELISA in PDL cells and macrophages","pmids":["26399972"],"confidence":"High","gaps":["TLR binding site on ASPN not mapped","Whether ASPN acts as TLR antagonist or sequesters ligand unclear"]},{"year":2015,"claim":"Confirmed in vivo relevance of ASPN as an osteogenesis inhibitor in a skeletal repair model.","evidence":"rBMSC overexpression transplanted into rat critical-size skull defects with histology","pmids":["26031659"],"confidence":"Medium","gaps":["Overexpression model only","No loss-of-function comparison"]},{"year":2019,"claim":"Placed ASPN under transcriptional control of vitamin D signaling, identifying a VDRE that links 1,25(OH)2D3 to ASPN suppression and enhanced osteogenesis.","evidence":"ChIP and luciferase reporter assays in hPDLSCs","pmids":["31837573"],"confidence":"Medium","gaps":["Single-lab regulatory mapping","Other transcriptional regulators not addressed"]},{"year":2021,"claim":"Identified a pro-adipogenic function for ASPN, broadening its mesenchymal lineage role beyond bone.","evidence":"Plap-1 knockout mice, siRNA in 3T3-L1, and recombinant protein lipid accumulation assays","pmids":["33654143"],"confidence":"Medium","gaps":["Signaling pathway driving adipogenesis not defined","Relation to BMP/FGF activities unclear"]},{"year":2022,"claim":"Established a hypoxia feedback loop in which HIF-1α induces ASPN, which in turn suppresses HIF-1α transcriptional activity.","evidence":"Hypoxic culture, chetomin inhibition, HRE reporter, and nuclear HIF-1α western blot","pmids":["35138637"],"confidence":"Medium","gaps":["Mechanism of HIF-1α suppression not defined","Limited orthogonal validation"]},{"year":2022,"claim":"Demonstrated through genetic ablation that ASPN is required for normal periodontal ligament collagen organization and protects against periodontitis-associated bone loss.","evidence":"PLAP-1 KO mice with micro-CT, TEM, histology, tooth extraction force, and ligature-induced periodontitis","pmids":["37958972"],"confidence":"High","gaps":["Molecular link between ASPN and collagen fibrillogenesis not detailed","Tissue-specific roles outside PDL untested"]},{"year":2022,"claim":"Implicated ASPN in cancer drug resistance, showing its knockdown reverses oxaliplatin resistance in colorectal cancer.","evidence":"siRNA knockdown with apoptosis assays and patient-derived xenograft model","pmids":["36228517"],"confidence":"Medium","gaps":["Mechanism of resistance not defined","No link to ASPN signaling partners"]},{"year":2023,"claim":"Identified HAPLN1 as an ASPN partner cooperating to inhibit osteogenic differentiation.","evidence":"Co-IP/binding and synergistic siRNA knockdown in BMSCs from OVX mice","pmids":["37427673"],"confidence":"Medium","gaps":["Interaction interface not mapped","Single-lab interaction data"]},{"year":2024,"claim":"Linked ASPN to gastric cancer progression via a MATN3-ASPN interaction amplifying tumor growth and metastasis.","evidence":"Protein-protein interaction analysis with overexpression/knockdown functional assays and in vivo tumor model","pmids":["39301785"],"confidence":"Medium","gaps":["Downstream signaling of MATN3-ASPN not defined","Mechanism of metastatic enhancement unclear"]},{"year":2026,"claim":"Showed ASPN transcription is driven by an exosomal NamiRNA acting on its super-enhancer during H. pylori infection, connecting ASPN to gastric inflammation.","evidence":"miRNA-seq, co-culture, H3K27ac ChIP, and in vivo antagomir treatment","pmids":["41723544"],"confidence":"Medium","gaps":["Single-lab epigenetic mechanism","Direct ASPN protein function in this context not tested"]},{"year":null,"claim":"How ASPN integrates its opposing activities (BMP-2 inhibition versus FGF-2 promotion) at the level of shared structural domains and matrix context, and how these reconcile with its roles in adipogenesis, fibrosis, and cancer, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of ASPN bound to any ligand","Unclear whether matrix sequestration vs receptor competition predominates across contexts"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,3,4,7]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[8]}],"localization":[{"term_id":"GO:0031012","term_label":"extracellular matrix","supporting_discovery_ids":[8]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[0]}],"pathway":[],"complexes":[],"partners":["BMP2","BMPR1B","FGF2","FGFR1","TLR2","TLR4","HAPLN1","MATN3"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BXN1","full_name":"Asporin","aliases":["Periodontal ligament-associated protein 1","PLAP-1"],"length_aa":380,"mass_kda":43.4,"function":"Negatively regulates periodontal ligament (PDL) differentiation and mineralization to ensure that the PDL is not ossified and to maintain homeostasis of the tooth-supporting system. Inhibits BMP2-induced cytodifferentiation of PDL cells by preventing its binding to BMPR1B/BMP type-1B receptor, resulting in inhibition of BMP-dependent activation of SMAD proteins (By similarity). Critical regulator of TGF-beta in articular cartilage and plays an essential role in cartilage homeostasis and osteoarthritis (OA) pathogenesis. Negatively regulates chondrogenesis in the articular cartilage by blocking the TGF-beta/receptor interaction on the cell surface and inhibiting the canonical TGF-beta/Smad signal. Binds calcium and plays a role in osteoblast-driven collagen biomineralization activity","subcellular_location":"Secreted, extracellular space, extracellular matrix","url":"https://www.uniprot.org/uniprotkb/Q9BXN1/entry"},"depmap":{"release":"DepMap","has_data":false,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ASPN"},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ASPN","total_profiled":1310},"omim":[{"mim_id":"608135","title":"ASPORIN; ASPN","url":"https://www.omim.org/entry/608135"},{"mim_id":"607850","title":"OSTEOARTHRITIS SUSCEPTIBILITY 3; OS3","url":"https://www.omim.org/entry/607850"},{"mim_id":"603932","title":"INTERVERTEBRAL DISC DISEASE; IDD","url":"https://www.omim.org/entry/603932"},{"mim_id":"165720","title":"OSTEOARTHRITIS SUSCEPTIBILITY 1; OS1","url":"https://www.omim.org/entry/165720"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Uncertain","locations":[{"location":"Nucleoplasm","reliability":"Uncertain"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"blood vessel","ntpm":129.3},{"tissue":"heart muscle","ntpm":89.2},{"tissue":"smooth muscle","ntpm":103.1}],"url":"https://www.proteinatlas.org/search/ASPN"},"hgnc":{"alias_symbol":["FLJ20129","SLRR1C","PLAP-1"],"prev_symbol":[]},"alphafold":{"accession":"Q9BXN1","domains":[{"cath_id":"3.80.10.10","chopping":"77-160","consensus_level":"medium","plddt":96.4682,"start":77,"end":160},{"cath_id":"3.80.10.10","chopping":"184-380","consensus_level":"medium","plddt":95.2692,"start":184,"end":380}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BXN1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BXN1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BXN1-F1-predicted_aligned_error_v6.png","plddt_mean":85.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ASPN","jax_strain_url":"https://www.jax.org/strain/search?query=ASPN"},"sequence":{"accession":"Q9BXN1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BXN1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BXN1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BXN1"}},"corpus_meta":[{"pmid":"17522060","id":"PMC_17522060","title":"PLAP-1/asporin, a novel negative regulator of periodontal ligament mineralization.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17522060","citation_count":160,"is_preprint":false},{"pmid":"20712325","id":"PMC_20712325","title":"Analysis of isoaspartic Acid by selective proteolysis with Asp-N and electron transfer dissociation mass spectrometry.","date":"2010","source":"Analytical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20712325","citation_count":88,"is_preprint":false},{"pmid":"22514560","id":"PMC_22514560","title":"ASPN and GJB2 Are Implicated in the Mechanisms of Invasion of Ductal Breast Carcinomas.","date":"2012","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/22514560","citation_count":70,"is_preprint":false},{"pmid":"21329514","id":"PMC_21329514","title":"Association of the D repeat polymorphism in the ASPN gene with developmental dysplasia of the hip: a case-control study in Han Chinese.","date":"2011","source":"Arthritis research & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/21329514","citation_count":60,"is_preprint":false},{"pmid":"17517696","id":"PMC_17517696","title":"Meta-analysis of association between the ASPN D-repeat and osteoarthritis.","date":"2007","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/17517696","citation_count":58,"is_preprint":false},{"pmid":"18407830","id":"PMC_18407830","title":"PLAP-1/asporin inhibits activation of BMP receptor via its leucine-rich repeat motif.","date":"2008","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/18407830","citation_count":49,"is_preprint":false},{"pmid":"22367347","id":"PMC_22367347","title":"miR-21 and miR-101 regulate PLAP-1 expression in periodontal ligament cells.","date":"2012","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/22367347","citation_count":48,"is_preprint":false},{"pmid":"2669754","id":"PMC_2669754","title":"Specificity of endoproteinase Asp-N (Pseudomonas fragi): cleavage at glutamyl residues in two proteins.","date":"1989","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/2669754","citation_count":37,"is_preprint":false},{"pmid":"16632759","id":"PMC_16632759","title":"Regulation of PLAP-1 expression in periodontal ligament cells.","date":"2006","source":"Journal of dental research","url":"https://pubmed.ncbi.nlm.nih.gov/16632759","citation_count":35,"is_preprint":false},{"pmid":"12944379","id":"PMC_12944379","title":"A method for the detection of asparagine deamidation and aspartate isomerization of proteins by MALDI/TOF-mass spectrometry using endoproteinase Asp-N.","date":"2003","source":"Journal of biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12944379","citation_count":32,"is_preprint":false},{"pmid":"26399972","id":"PMC_26399972","title":"PLAP-1/Asporin Regulates TLR2- and TLR4-induced Inflammatory Responses.","date":"2015","source":"Journal of dental research","url":"https://pubmed.ncbi.nlm.nih.gov/26399972","citation_count":30,"is_preprint":false},{"pmid":"36228517","id":"PMC_36228517","title":"Precision medicine-guided co-delivery of ASPN siRNA and oxaliplatin by nanoparticles to overcome chemoresistance of colorectal cancer.","date":"2022","source":"Biomaterials","url":"https://pubmed.ncbi.nlm.nih.gov/36228517","citation_count":26,"is_preprint":false},{"pmid":"24453179","id":"PMC_24453179","title":"Inhibitory effects of PLAP-1/asporin on periodontal ligament cells.","date":"2014","source":"Journal of dental research","url":"https://pubmed.ncbi.nlm.nih.gov/24453179","citation_count":25,"is_preprint":false},{"pmid":"26239644","id":"PMC_26239644","title":"PLAP-1/Asporin Positively Regulates FGF-2 Activity.","date":"2015","source":"Journal of dental research","url":"https://pubmed.ncbi.nlm.nih.gov/26239644","citation_count":23,"is_preprint":false},{"pmid":"36245218","id":"PMC_36245218","title":"Plap-1 lineage tracing and single-cell transcriptomics reveal cellular dynamics in the periodontal ligament.","date":"2022","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/36245218","citation_count":23,"is_preprint":false},{"pmid":"31665048","id":"PMC_31665048","title":"Identifying the role of ASPN and COMP genes in knee osteoarthritis development.","date":"2019","source":"Journal of orthopaedic surgery and research","url":"https://pubmed.ncbi.nlm.nih.gov/31665048","citation_count":23,"is_preprint":false},{"pmid":"16311710","id":"PMC_16311710","title":"High-resolution SNP map of ASPN, a susceptibility gene for osteoarthritis.","date":"2005","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16311710","citation_count":18,"is_preprint":false},{"pmid":"29551890","id":"PMC_29551890","title":"Synovial tissue quantitative proteomics analysis reveals paeoniflorin decreases LIFR and ASPN proteins in experimental rheumatoid arthritis.","date":"2018","source":"Drug design, development and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/29551890","citation_count":17,"is_preprint":false},{"pmid":"33533513","id":"PMC_33533513","title":"LncRNA DCST1-AS1 inhibits PDLCs' proliferation in periodontitis and may bind with miR-21 precursor to upregulate PLAP-1.","date":"2021","source":"Journal of periodontal research","url":"https://pubmed.ncbi.nlm.nih.gov/33533513","citation_count":16,"is_preprint":false},{"pmid":"25038933","id":"PMC_25038933","title":"Overexpression of the PLAP-1 gene inhibits the differentiation of BMSCs into osteoblast-like cells.","date":"2014","source":"Journal of molecular histology","url":"https://pubmed.ncbi.nlm.nih.gov/25038933","citation_count":14,"is_preprint":false},{"pmid":"31837573","id":"PMC_31837573","title":"1,25(OH)2D3 supports the osteogenic differentiation of hPDLSCs under inflammatory conditions through inhibiting PLAP-1 expression transcriptionally.","date":"2019","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/31837573","citation_count":14,"is_preprint":false},{"pmid":"33654143","id":"PMC_33654143","title":"Mice lacking PLAP-1/asporin counteracts high fat diet-induced metabolic disorder and alveolar bone loss by controlling adipose tissue expansion.","date":"2021","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33654143","citation_count":14,"is_preprint":false},{"pmid":"34663368","id":"PMC_34663368","title":"MiR-4303 relieves chondrocyte inflammation by targeting ASPN in osteoarthritis.","date":"2021","source":"Journal of orthopaedic surgery and research","url":"https://pubmed.ncbi.nlm.nih.gov/34663368","citation_count":14,"is_preprint":false},{"pmid":"28747338","id":"PMC_28747338","title":"Copy number loss in the region of the ASPN gene in patients with acetabular dysplasia: ASPN CNV in acetabular dysplasia.","date":"2017","source":"Bone & joint research","url":"https://pubmed.ncbi.nlm.nih.gov/28747338","citation_count":14,"is_preprint":false},{"pmid":"1981672","id":"PMC_1981672","title":"Relaxed specificity of endoproteinase Asp-N: this enzyme cleaves at peptide bonds N-terminal to glutamate as well as aspartate and cysteic acid residues.","date":"1990","source":"Biochemistry international","url":"https://pubmed.ncbi.nlm.nih.gov/1981672","citation_count":14,"is_preprint":false},{"pmid":"25030405","id":"PMC_25030405","title":"Association between single nucleotide polymorphisms of asporin (ASPN) and BMP5 with the risk of knee osteoarthritis in a Chinese Han population.","date":"2014","source":"Cell biochemistry and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/25030405","citation_count":10,"is_preprint":false},{"pmid":"23733110","id":"PMC_23733110","title":"The D-repeat polymorphism in the ASPN gene and primary knee osteoarthritis in a Mexican mestizo population: a case-control study.","date":"2013","source":"Journal of orthopaedic science : official journal of the Japanese Orthopaedic Association","url":"https://pubmed.ncbi.nlm.nih.gov/23733110","citation_count":9,"is_preprint":false},{"pmid":"16879361","id":"PMC_16879361","title":"Porcine OGN and ASPN: mapping, polymorphisms and use for quantitative trait loci identification for growth and carcass traits in a Meishan x Piétrain intercross.","date":"2006","source":"Animal genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16879361","citation_count":9,"is_preprint":false},{"pmid":"36394011","id":"PMC_36394011","title":"Identification of nanoparticle-mediated siRNA-ASPN as a key gene target in the treatment of keloids.","date":"2022","source":"Frontiers in bioengineering and biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/36394011","citation_count":8,"is_preprint":false},{"pmid":"37427673","id":"PMC_37427673","title":"ASPN Synergizes with HAPLN1 to Inhibit the Osteogenic Differentiation of Bone Marrow Mesenchymal Stromal Cells and Extracellular Matrix Mineralization of Osteoblasts.","date":"2023","source":"Orthopaedic surgery","url":"https://pubmed.ncbi.nlm.nih.gov/37427673","citation_count":8,"is_preprint":false},{"pmid":"26031659","id":"PMC_26031659","title":"Overexpression of PLAP-1 in bone marrow stromal cells inhibits the rat critical-size skull defect repair.","date":"2015","source":"Journal of molecular histology","url":"https://pubmed.ncbi.nlm.nih.gov/26031659","citation_count":7,"is_preprint":false},{"pmid":"21146486","id":"PMC_21146486","title":"Selective isolation of N-blocked peptide by combining AspN digestion, transamination, and tosylhydrazine glass treatment.","date":"2010","source":"Analytical biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21146486","citation_count":7,"is_preprint":false},{"pmid":"35138637","id":"PMC_35138637","title":"Reciprocal role of PLAP-1 in HIF-1α-mediated responses to hypoxia.","date":"2022","source":"Journal of periodontal research","url":"https://pubmed.ncbi.nlm.nih.gov/35138637","citation_count":4,"is_preprint":false},{"pmid":"37958972","id":"PMC_37958972","title":"Mice Lacking PLAP-1/Asporin Show Alteration of Periodontal Ligament Structures and Acceleration of Bone Loss in Periodontitis.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/37958972","citation_count":4,"is_preprint":false},{"pmid":"39301785","id":"PMC_39301785","title":"Investigating MATN3 and ASPN as novel drivers of gastric cancer progression via EMT pathways.","date":"2024","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/39301785","citation_count":3,"is_preprint":false},{"pmid":"21431266","id":"PMC_21431266","title":"[Construction and confirmation of a recombinant eukaryotic expression plasmid pBABE-hygro-PLAP-1].","date":"2010","source":"Shanghai kou qiang yi xue = Shanghai journal of stomatology","url":"https://pubmed.ncbi.nlm.nih.gov/21431266","citation_count":3,"is_preprint":false},{"pmid":"26016288","id":"PMC_26016288","title":"[D-repeat polymorphism in the ASPN gene in knee osteoarthritis in females in Torreón, Coahuila. Case-control study].","date":"2014","source":"Acta ortopedica mexicana","url":"https://pubmed.ncbi.nlm.nih.gov/26016288","citation_count":3,"is_preprint":false},{"pmid":"39928346","id":"PMC_39928346","title":"Defect Imide Double Antiperovskites AE5AsPn(NH)2 (AE=Ca, Sr; Pn=Sb, Bi) as Potential Solar Cell Absorber Materials.","date":"2025","source":"Angewandte Chemie (International ed. in English)","url":"https://pubmed.ncbi.nlm.nih.gov/39928346","citation_count":3,"is_preprint":false},{"pmid":"39662603","id":"PMC_39662603","title":"Single-cell transcriptomic analysis and luteolin treatment reveal three adipogenic genes, including Aspn, Htra1 and Efemp1.","date":"2024","source":"Biochimica et biophysica acta. Molecular and cell biology of lipids","url":"https://pubmed.ncbi.nlm.nih.gov/39662603","citation_count":2,"is_preprint":false},{"pmid":"41258306","id":"PMC_41258306","title":"Exploring ASPN as a pan-cancer biomarker with a focus on gastric cancer.","date":"2025","source":"Discover oncology","url":"https://pubmed.ncbi.nlm.nih.gov/41258306","citation_count":0,"is_preprint":false},{"pmid":"41689014","id":"PMC_41689014","title":"Engineered Cas9 exosome vesicles as a novel gene editing tool for targeted ASPN editing in osteoarthritis.","date":"2026","source":"Journal of nanobiotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/41689014","citation_count":0,"is_preprint":false},{"pmid":"41723544","id":"PMC_41723544","title":"Epithelial cell-derived exosomes carry NamiRNA-143-5p and promote ASPN expression in fibroblasts to induce Helicobacter pylori infected gastritis progression.","date":"2026","source":"Gut pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/41723544","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.11.21.624638","title":"Unravelling the molecular activation of the reparative cardiac fibroblasts after myocardial infarction","date":"2024-11-21","source":"bioRxiv","url":"https://doi.org/10.1101/2024.11.21.624638","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":23614,"output_tokens":3800,"usd":0.063921,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11477,"output_tokens":4080,"usd":0.079692,"stage2_stop_reason":"end_turn"},"total_usd":0.143613,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"PLAP-1/asporin directly binds BMP-2, as demonstrated by co-immunoprecipitation, and negatively regulates BMP-2-induced cytodifferentiation and mineralization of periodontal ligament (PDL) cells; overexpression inhibited mineralization while knockdown enhanced BMP-2-induced differentiation.\",\n      \"method\": \"Co-immunoprecipitation, overexpression, RNAi knockdown, immunohistochemistry co-localization\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP plus gain- and loss-of-function experiments in the same study, replicated in subsequent papers\",\n      \"pmids\": [\"17522060\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"PLAP-1/asporin inhibits BMP-2 signaling by competitively preventing BMP-2 from binding to BMP receptor-IB (BMPR-IB), thereby blocking Smad activation; the leucine-rich repeat 5 (LRR5) motif is the functional domain mediating this interaction, as LRR5 mutation rescues inhibition and a 26-aa LRR5-derived peptide recapitulates inhibition.\",\n      \"method\": \"Recombinant protein competition binding assay, site-directed mutagenesis of LRR5, peptide inhibition assay, Smad phosphorylation western blot\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro receptor-binding competition assay combined with mutagenesis and peptide rescue in a single study\",\n      \"pmids\": [\"18407830\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"The aspartic acid (D) repeat polymorphism of PLAP-1/asporin influences its functional potency: D14-PLAP-1 suppresses BMP-2-induced cytodifferentiation more strongly than D13-PLAP-1 and shows stronger binding affinity to BMP-2 by co-immunoprecipitation.\",\n      \"method\": \"Stable transfection of D13 vs D14 cell lines, alkaline phosphatase and alizarin red staining, co-immunoprecipitation, western blot, luciferase reporter assay\",\n      \"journal\": \"Journal of dental research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus functional reporter and staining assays in single lab, multiple orthogonal methods\",\n      \"pmids\": [\"24453179\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"PLAP-1/asporin positively regulates FGF-2 activity by directly binding FGF-2 and promoting FGF-2–FGFR1 complex formation; Plap-1 knockout MEFs show defective FGF-2 responses rescued by Plap-1 transfection, and reduced FGF-2/FGFR1 co-localization.\",\n      \"method\": \"Plap-1 knockout mouse-derived MEFs, binding assay, immunocytochemistry co-localization, rescue transfection\",\n      \"journal\": \"Journal of dental research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with rescue, direct binding assay, and co-localization in a single study with multiple orthogonal methods\",\n      \"pmids\": [\"26239644\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"PLAP-1/asporin directly binds TLR2 and TLR4 (shown by immunoprecipitation), suppresses TLR2/4-induced NF-κB activity, reduces IκBα kinase degradation, and downregulates proinflammatory cytokine expression in PDL cells and macrophages.\",\n      \"method\": \"Overexpression, recombinant protein treatment, immunoprecipitation, NF-κB luciferase reporter, western blot for IκBα, cytokine ELISA\",\n      \"journal\": \"Journal of dental research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus multiple downstream functional readouts (NF-κB activity, IκBα, cytokines) in single lab with orthogonal methods\",\n      \"pmids\": [\"26399972\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PLAP-1/asporin enhances adipogenesis: recombinant PLAP-1 promotes lipid accumulation in 3T3-L1 cells, Plap-1 knockout mice and Plap-1-knockdown 3T3-L1 cells show reduced lipid accumulation, and primary preadipocytes from Plap-1 KO mice exhibit less adipogenic differentiation than wild-type.\",\n      \"method\": \"Plap-1 knockout mouse model, siRNA knockdown in 3T3-L1 cells, recombinant protein treatment, lipid staining/accumulation assays\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO plus siRNA and recombinant protein gain-of-function in single lab\",\n      \"pmids\": [\"33654143\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"1,25(OH)2D3 suppresses PLAP-1 expression transcriptionally through a vitamin D response element (VDRE) in the PLAP-1 promoter that binds VDR, as confirmed by ChIP assay and reporter gene assays, leading to enhanced osteogenic differentiation of hPDLSCs under inflammatory conditions.\",\n      \"method\": \"ChIP assay, luciferase reporter assay, western blot, RT-qPCR\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus reporter gene assay identifying specific VDRE, single lab with two orthogonal methods\",\n      \"pmids\": [\"31837573\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PLAP-1/asporin suppresses HIF-1α signaling: hypoxia-induced PLAP-1 expression is HIF-1α-dependent (blocked by chetomin), and recombinant PLAP-1 or PLAP-1 gene transfection reduces hypoxia-induced HRE-luciferase activity and nuclear HIF-1α accumulation in PDL cells.\",\n      \"method\": \"Hypoxic cell culture, chemical inhibitors, luciferase HRE reporter assay, western blot for nuclear HIF-1α, recombinant protein and transfection experiments\",\n      \"journal\": \"Journal of periodontal research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — reporter assay and western blot in single lab, multiple conditions but limited orthogonal validation\",\n      \"pmids\": [\"35138637\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PLAP-1/asporin knockout mice display enlarged periodontal ligament space, increased collagen fibril diameter, altered ECM protein expression (elevated Col3, BGN, DCN), reduced tooth extraction force, and accelerated alveolar bone resorption with more osteoclasts in ligature-induced periodontitis, establishing a structural and protective role for PLAP-1 in PDL collagen organization and periodontal inflammation.\",\n      \"method\": \"PLAP-1 KO mouse model, micro-CT, histology (HE, picrosirius red), fluorescence immunostaining, TEM, tooth extraction force measurement, ligature-induced periodontitis model\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with multiple orthogonal structural and functional readouts in vivo\",\n      \"pmids\": [\"37958972\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Overexpression of PLAP-1 in rat bone marrow stromal cells (rBMSCs) inhibits their differentiation into osteoblast-like cells, reducing mineralized nodule formation and decreasing expression of osteoblast markers (Runx2, Osterix, ALP, BSP, osteocalcin).\",\n      \"method\": \"Stable retroviral transfection of PLAP-1 in rBMSCs, alizarin red staining, RT-qPCR for osteogenic markers\",\n      \"journal\": \"Journal of molecular histology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — gain-of-function in cell culture with multiple marker readouts, single lab\",\n      \"pmids\": [\"25038933\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Overexpression of PLAP-1 in rBMSCs transplanted into rat critical-size skull defects inhibits new bone formation and mineralization in vivo, confirming its role as a negative regulator of osteogenesis in a skeletal repair context.\",\n      \"method\": \"In vivo rat skull defect model, rBMSC transplantation, X-ray, HE/Masson/von Kossa histology, immunohistochemistry\",\n      \"journal\": \"Journal of molecular histology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — in vivo KO/OE model with histological readouts, single lab\",\n      \"pmids\": [\"26031659\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ASPN interacts with HAPLN1, and their combined knockdown synergistically increases ALP, OPN, OCN, and COL1A1 expression and ECM mineralization in BMSCs while decreasing osteoclast markers in bone marrow macrophages, indicating ASPN and HAPLN1 cooperate to inhibit osteogenic differentiation.\",\n      \"method\": \"Co-IP/binding analysis, siRNA knockdown in BMSCs from OVX mice, western blot for osteogenic and osteoclast markers, ECM mineralization assay\",\n      \"journal\": \"Orthopaedic surgery\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — protein interaction analysis plus siRNA functional assays in single lab\",\n      \"pmids\": [\"37427673\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ASPN mediates oxaliplatin (OXA) resistance in colorectal cancer; siRNA-mediated ASPN knockdown reverses OXA resistance and promotes cell apoptosis both in vitro and in patient-derived xenograft models in vivo.\",\n      \"method\": \"siRNA knockdown, in vitro cell viability/apoptosis assays, patient-derived xenograft (PDX) mouse model, nanoparticle co-delivery system\",\n      \"journal\": \"Biomaterials\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined phenotypic readout in vitro and PDX in vivo, single lab\",\n      \"pmids\": [\"36228517\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Exosomal miR-143-5p from H. pylori-infected epithelial cells functions as a nuclear activating microRNA (NamiRNA) that binds the super-enhancer region of the ASPN gene, increases H3K27ac enrichment, and promotes ASPN transcription in fibroblasts, leading to upregulation of pro-inflammatory cytokines (IL-4, IL-6, TGF-β); in vivo antagomir-143-5p reduced ASPN and cytokine expression and alleviated gastric inflammation.\",\n      \"method\": \"MicroRNA sequencing, co-culture assay, immunofluorescence, ChIP for H3K27ac, in vivo antagomir treatment\",\n      \"journal\": \"Gut pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus co-culture, in vivo antagomir, and immunofluorescence in single lab with orthogonal methods\",\n      \"pmids\": [\"41723544\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ASPN overexpression amplifies MATN3-driven gastric cancer cell proliferation, migration, and invasion, and MATN3-ASPN protein-protein interaction was confirmed; co-overexpression of MATN3 and ASPN enhanced tumor growth and metastasis in vivo.\",\n      \"method\": \"Protein-protein interaction analysis, co-expression analysis, overexpression/knockdown functional assays (proliferation, migration, invasion, apoptosis), in vivo mouse tumor model\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — protein interaction plus in vitro and in vivo functional assays, single lab\",\n      \"pmids\": [\"39301785\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ASPN/PLAP-1 is an extracellular matrix small leucine-rich repeat proteoglycan that acts as a multi-target negative regulator of growth factor signaling: it directly binds BMP-2 via its LRR5 motif to competitively block BMPR-IB engagement and downstream Smad activation, directly binds TGF-β1 to inhibit TGF-β/Smad signaling, and directly binds FGF-2 to promote FGF-2–FGFR1 complex formation (a positive regulatory role); it also binds TLR2 and TLR4 to suppress NF-κB-mediated inflammatory cytokine production, suppresses HIF-1α transcriptional activity in a feedback loop, regulates collagen fibril organization in the periodontal ligament, promotes adipogenesis, and inhibits osteogenic differentiation of PDL and bone marrow stromal cells, with the N-terminal aspartic acid repeat polymorphism modulating the strength of BMP-2 binding and inhibition.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ASPN (PLAP-1/asporin) is an extracellular matrix small leucine-rich repeat proteoglycan that acts as a multi-target regulator of growth factor and inflammatory signaling and as a structural organizer of connective tissue [#0, #8]. It directly binds BMP-2 and inhibits BMP-2-induced cytodifferentiation and mineralization, competitively blocking BMP-2 engagement of BMP receptor-IB and the downstream Smad cascade through its LRR5 motif, which alone is sufficient to recapitulate inhibition [#0, #1]; the N-terminal aspartic acid repeat length tunes this activity, with the D14 variant binding BMP-2 more strongly and suppressing differentiation more potently than D13 [#2]. In contrast to its inhibitory role toward BMP-2, ASPN positively regulates FGF-2 by binding it and promoting FGF-2\\u2013FGFR1 complex formation [#3]. ASPN also binds TLR2 and TLR4 to dampen NF-\\u03baB activation, stabilizing I\\u03baB\\u03b1 and reducing proinflammatory cytokine output [#4], and it suppresses HIF-1\\u03b1 transcriptional activity in a hypoxia-induced feedback loop [#7]. Consistent with a matrix-organizing function, ASPN-deficient mice show enlarged periodontal ligament space, altered collagen fibril architecture, and accelerated alveolar bone resorption [#8]. Across mesenchymal lineages ASPN restrains osteogenic differentiation of periodontal ligament and bone marrow stromal cells while promoting adipogenesis [#5, #9, #10]. In cancer, ASPN contributes to oxaliplatin resistance in colorectal cancer and amplifies MATN3-driven gastric cancer progression [#12, #14].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established ASPN as a direct binding partner and negative regulator of BMP-2 signaling, defining its first molecular function beyond being a matrix proteoglycan.\",\n      \"evidence\": \"Co-immunoprecipitation with gain- and loss-of-function in periodontal ligament cells\",\n      \"pmids\": [\"17522060\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve the binding domain or whether inhibition was receptor-competitive\", \"Limited to PDL cell context\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Resolved the mechanism of BMP-2 inhibition as competitive blockade of BMPR-IB and mapped the activity to the LRR5 motif, converting a binding observation into a defined structure-function model.\",\n      \"evidence\": \"Recombinant competition binding, LRR5 mutagenesis, and a 26-aa peptide rescue with Smad phosphorylation readout\",\n      \"pmids\": [\"18407830\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural model of the ASPN\\u2013BMP-2 interface\", \"Relevance in vivo not tested\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Showed the N-terminal aspartic acid repeat polymorphism tunes ASPN function, linking an allelic variant to differential BMP-2 binding strength and inhibitory potency.\",\n      \"evidence\": \"D13 vs D14 stable cell lines, Co-IP, reporter and mineralization assays\",\n      \"pmids\": [\"24453179\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab Co-IP for affinity difference\", \"Molecular basis for repeat-dependent affinity not defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Extended the anti-osteogenic role beyond PDL by showing ASPN inhibits osteoblast differentiation of bone marrow stromal cells.\",\n      \"evidence\": \"Retroviral overexpression in rBMSCs with osteogenic marker and mineralization assays\",\n      \"pmids\": [\"25038933\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Gain-of-function only\", \"Mechanism downstream of BMP/FGF not separated\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Revealed a context-dependent positive regulatory role: ASPN binds FGF-2 and promotes FGF-2\\u2013FGFR1 complex formation, contrasting with its BMP-2 inhibition.\",\n      \"evidence\": \"Plap-1 knockout MEFs with rescue, binding assay, and FGF-2/FGFR1 co-localization\",\n      \"pmids\": [\"26239644\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How ASPN can be inhibitory for BMP-2 yet stimulatory for FGF-2 mechanistically unresolved\", \"Domain mediating FGF-2 binding not mapped\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Defined an anti-inflammatory function through direct TLR2/TLR4 binding and suppression of NF-\\u03baB signaling.\",\n      \"evidence\": \"Immunoprecipitation, NF-\\u03baB reporter, I\\u03baB\\u03b1 western blot, cytokine ELISA in PDL cells and macrophages\",\n      \"pmids\": [\"26399972\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"TLR binding site on ASPN not mapped\", \"Whether ASPN acts as TLR antagonist or sequesters ligand unclear\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Confirmed in vivo relevance of ASPN as an osteogenesis inhibitor in a skeletal repair model.\",\n      \"evidence\": \"rBMSC overexpression transplanted into rat critical-size skull defects with histology\",\n      \"pmids\": [\"26031659\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Overexpression model only\", \"No loss-of-function comparison\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Placed ASPN under transcriptional control of vitamin D signaling, identifying a VDRE that links 1,25(OH)2D3 to ASPN suppression and enhanced osteogenesis.\",\n      \"evidence\": \"ChIP and luciferase reporter assays in hPDLSCs\",\n      \"pmids\": [\"31837573\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab regulatory mapping\", \"Other transcriptional regulators not addressed\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified a pro-adipogenic function for ASPN, broadening its mesenchymal lineage role beyond bone.\",\n      \"evidence\": \"Plap-1 knockout mice, siRNA in 3T3-L1, and recombinant protein lipid accumulation assays\",\n      \"pmids\": [\"33654143\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Signaling pathway driving adipogenesis not defined\", \"Relation to BMP/FGF activities unclear\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Established a hypoxia feedback loop in which HIF-1\\u03b1 induces ASPN, which in turn suppresses HIF-1\\u03b1 transcriptional activity.\",\n      \"evidence\": \"Hypoxic culture, chetomin inhibition, HRE reporter, and nuclear HIF-1\\u03b1 western blot\",\n      \"pmids\": [\"35138637\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of HIF-1\\u03b1 suppression not defined\", \"Limited orthogonal validation\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrated through genetic ablation that ASPN is required for normal periodontal ligament collagen organization and protects against periodontitis-associated bone loss.\",\n      \"evidence\": \"PLAP-1 KO mice with micro-CT, TEM, histology, tooth extraction force, and ligature-induced periodontitis\",\n      \"pmids\": [\"37958972\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular link between ASPN and collagen fibrillogenesis not detailed\", \"Tissue-specific roles outside PDL untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Implicated ASPN in cancer drug resistance, showing its knockdown reverses oxaliplatin resistance in colorectal cancer.\",\n      \"evidence\": \"siRNA knockdown with apoptosis assays and patient-derived xenograft model\",\n      \"pmids\": [\"36228517\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of resistance not defined\", \"No link to ASPN signaling partners\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified HAPLN1 as an ASPN partner cooperating to inhibit osteogenic differentiation.\",\n      \"evidence\": \"Co-IP/binding and synergistic siRNA knockdown in BMSCs from OVX mice\",\n      \"pmids\": [\"37427673\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interaction interface not mapped\", \"Single-lab interaction data\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Linked ASPN to gastric cancer progression via a MATN3-ASPN interaction amplifying tumor growth and metastasis.\",\n      \"evidence\": \"Protein-protein interaction analysis with overexpression/knockdown functional assays and in vivo tumor model\",\n      \"pmids\": [\"39301785\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Downstream signaling of MATN3-ASPN not defined\", \"Mechanism of metastatic enhancement unclear\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showed ASPN transcription is driven by an exosomal NamiRNA acting on its super-enhancer during H. pylori infection, connecting ASPN to gastric inflammation.\",\n      \"evidence\": \"miRNA-seq, co-culture, H3K27ac ChIP, and in vivo antagomir treatment\",\n      \"pmids\": [\"41723544\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab epigenetic mechanism\", \"Direct ASPN protein function in this context not tested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How ASPN integrates its opposing activities (BMP-2 inhibition versus FGF-2 promotion) at the level of shared structural domains and matrix context, and how these reconcile with its roles in adipogenesis, fibrosis, and cancer, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of ASPN bound to any ligand\", \"Unclear whether matrix sequestration vs receptor competition predominates across contexts\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 3, 4, 7]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0031012\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0162582\", \"supporting_discovery_ids\": [0, 1, 3, 4]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"BMP2\", \"BMPR1B\", \"FGF2\", \"FGFR1\", \"TLR2\", \"TLR4\", \"HAPLN1\", \"MATN3\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}