{"gene":"CENPM","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2014,"finding":"CENP-M is structurally and evolutionarily related to small GTPases but is incapable of GTP-binding and conformational switching (pseudo GTPase). CENP-M is crucially required for the assembly and stability of the HIKM tetramer comprising CENP-I, CENP-H, and CENP-K. A point mutant disrupting the CENP-M/CENP-I interaction hampers kinetochore assembly and chromosome alignment, and prevents kinetochore recruitment of the CENP-T/W complex, placing CENP-M downstream of CENP-C in a single kinetochore assembly pathway.","method":"Crystal structure, in vitro biochemical reconstitution, Co-IP/pulldown, active-site/interface mutagenesis, live-cell imaging, cell biological loss-of-function assays","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure combined with biochemical reconstitution, mutagenesis, and cell biological validation in a single rigorous study","pmids":["25006165"],"is_preprint":false},{"year":2006,"finding":"An alternative transcript of the PANE1/CENPM gene encodes a novel HLA-A*0301-restricted minor histocompatibility antigen (mHAg) selectively expressed in resting CD19+ B cells and B-CLL cells. The antigenic peptide is encoded within a unique exon, and a single nucleotide polymorphism replacing an arginine codon with a stop codon determines differential T-cell recognition. Activation of B-CLL cells via CD40L decreases expression of the mHAg-encoding transcript and reciprocally increases expression of transcripts lacking this exon.","method":"Sequencing of PANE1 alleles, mass spectrometry peptide identification, T-cell recognition assays, RT-PCR expression analysis","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (MS, sequencing, T-cell assay) in a single lab establishing isoform-specific antigen presentation","pmids":["16391015"],"is_preprint":false},{"year":2019,"finding":"CENPM knockdown in hepatocellular carcinoma cells inhibits cell proliferation in vitro and in vivo, suppresses migration and invasion, promotes apoptosis, and arrests the cell cycle. GSEA and functional validation linked CENPM activity to the p53 signaling pathway and cell cycle pathway. miR-1270 was identified as a negative post-transcriptional regulator of CENPM.","method":"siRNA knockdown, xenograft in vivo model, Western blot, flow cytometry, GSEA bioinformatics, miRNA luciferase reporter assay","journal":"Journal of experimental & clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined cellular phenotypes and pathway placement via GSEA plus miRNA reporter assay, single lab","pmids":["31703591"],"is_preprint":false},{"year":2020,"finding":"CENPM knockdown in pancreatic cancer cells inhibits cell proliferation, alters the cell cycle, and limits cell migration and invasion via the mTOR/p70S6K signaling pathway.","method":"siRNA knockdown, cell proliferation assays, cell cycle analysis, migration/invasion assays, Western blot for mTOR/p70S6K pathway components","journal":"Oncology reports","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement by Western blot without pharmacological rescue or upstream mechanistic detail","pmids":["32705259"],"is_preprint":false},{"year":2022,"finding":"CENPM promotes lung adenocarcinoma cell cycle progression, proliferation, migration and invasion by activating phosphorylation of mTOR (not by altering total mTOR protein levels). Inhibition of mTOR activity abrogates the promoting effects of CENPM overexpression.","method":"Gain/loss-of-function assays, Western blot for mTOR phosphorylation, mTOR inhibitor rescue experiment, xenograft model","journal":"Acta biochimica et biophysica Sinica","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement supported by inhibitor rescue but no upstream mechanistic detail of how CENPM activates mTOR","pmids":["35130633"],"is_preprint":false},{"year":2022,"finding":"HPV E5 oncoprotein enhances CENPM expression through the transcription factor E2F1 binding to the CENPM promoter in head and neck squamous cell carcinoma. Inhibition of CENPM expression in HPV-positive HNSCC cells increases resistance to X-ray radiation (~59% under 2 Gy), associated with a reduced proportion of mitotic cells.","method":"ChIP assay for E2F1 binding at CENPM promoter, siRNA knockdown, CRISPR/Cas9 models, Western blot, qRT-PCR, clonogenic survival assay","journal":"Oral oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP directly demonstrates E2F1 binding to CENPM promoter; functional consequence validated by knockdown and CRISPR in single lab","pmids":["35462155"],"is_preprint":false},{"year":2024,"finding":"E2F1 acts as a key transcriptional regulator of CENPM in glioblastoma. CENPM promotes GBM cell proliferation, invasion, and glycolytic reprogramming (glucose consumption, lactate production, ATP levels). The glycolytic inhibitor 2-DG reverses CENPM overexpression effects, indicating dependence on glycolytic pathways. In vivo, CENPM knockdown reduced tumor volume by ~60%.","method":"shRNA knockdown, vector-based overexpression, glycolytic metabolite measurement, 2-DG pharmacological rescue, xenograft mouse model","journal":"Cell biology and toxicology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement by rescue experiment but mechanistic link between CENPM and glycolysis is not biochemically characterized","pmids":["39707034"],"is_preprint":false},{"year":2024,"finding":"CENPM knockdown in ovarian cancer cells activates the cGAS-STING pathway and promotes pyroptosis, suppressing proliferation, migration and invasion. A cGAS-STING pathway inhibitor reversed the inhibitory effects of CENPM knockdown on viability, migration and invasion, placing CENPM upstream of cGAS-STING in ovarian cancer cells.","method":"siRNA knockdown, pharmacological pathway inhibitor rescue, ELISA for inflammatory factors, Western blot, xenograft model","journal":"BMC cancer","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement supported by inhibitor rescue but no direct biochemical link between CENPM and cGAS-STING activation","pmids":["38693472"],"is_preprint":false},{"year":2025,"finding":"CENPM physically interacts with immune checkpoint ligand FGL1 in adrenocortical carcinoma cells. CENPM knockdown downregulates FGL1, suppresses ECM-related collagen signaling, and inhibits ACC cell invasion and migration; overexpression of FGL1 rescues the migration and invasion defects caused by CENPM knockdown.","method":"Co-immunoprecipitation, DIA quantitative proteomics, immunofluorescence colocalization, Western blot, rescue overexpression experiment, xenograft NPG mice","journal":"Clinical and translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — physical interaction validated by Co-IP plus proteomics, functional consequence confirmed by rescue assay, single lab","pmids":["39778025"],"is_preprint":false},{"year":2025,"finding":"CENPM knockdown in glioma cells induces G0/G1 phase cell cycle arrest and inhibits proliferation. RNA-seq and Western blot analyses demonstrate that CENPM activates the PI3K/AKT signaling pathway in glioma cells; in vivo knockdown reduces tumor growth.","method":"shRNA knockdown, RNA-seq, Western blot for PI3K/AKT pathway, cell cycle flow cytometry, xenograft model","journal":"Genomics","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement by Western blot and RNA-seq without upstream mechanistic detail or rescue","pmids":["41317748"],"is_preprint":false}],"current_model":"CENPM (CENP-M) is a pseudo GTPase that is structurally related to small GTPases but cannot bind GTP; it is essential for assembling the CENP-H/I/K/M (HIKM) tetramer at the kinetochore–centromere interface downstream of CENP-C, thereby enabling CENP-T/W recruitment and faithful chromosome segregation during mitosis. In cancer contexts, CENPM expression is transcriptionally driven by E2F1 (and HPV E5 via E2F1), and its activity has been functionally linked to mTOR/p70S6K and PI3K/AKT signaling, glycolytic reprogramming, and physical interaction with the immune checkpoint ligand FGL1, though the biochemical mechanisms connecting CENPM to these non-kinetochore pathways remain incompletely characterized."},"narrative":{"mechanistic_narrative":"CENPM (CENP-M) is a centromere-associated protein essential for kinetochore assembly and faithful chromosome segregation during mitosis [PMID:25006165]. Structurally and evolutionarily related to small GTPases, CENP-M is a pseudo GTPase that cannot bind GTP or undergo conformational switching; instead it functions as a scaffolding subunit required for the assembly and stability of the HIKM tetramer with CENP-I, CENP-H, and CENP-K [PMID:25006165]. Disruption of the CENP-M/CENP-I interface impairs kinetochore assembly and chromosome alignment and blocks recruitment of the CENP-T/W complex, placing CENP-M in a single kinetochore assembly pathway downstream of CENP-C [PMID:25006165]. An alternative transcript of the CENPM/PANE1 locus encodes an HLA-A*0301-restricted minor histocompatibility antigen selectively expressed in resting B cells and B-CLL cells, with a stop-codon polymorphism determining T-cell recognition [PMID:16391015]. In cancer, CENPM expression is transcriptionally driven by E2F1, including induction by the HPV E5 oncoprotein via E2F1 binding to the CENPM promoter, where it supports mitotic progression and radiosensitivity [PMID:35462155]. CENPM physically interacts with the immune checkpoint ligand FGL1 and sustains FGL1 levels and ECM-related collagen signaling to promote tumor cell invasion and migration [PMID:39778025].","teleology":[{"year":2006,"claim":"Established that the CENPM/PANE1 locus produces an alternative transcript encoding a tissue-restricted minor histocompatibility antigen, defining a non-mitotic, immunologically relevant output of the gene.","evidence":"Allele sequencing, mass spectrometry peptide identification, T-cell recognition and RT-PCR assays in B-CLL cells","pmids":["16391015"],"confidence":"Medium","gaps":["Does not connect the antigen-encoding isoform to the canonical centromeric function of CENP-M","Functional role of the alternative protein product beyond antigen presentation is unknown"]},{"year":2014,"claim":"Defined the core molecular function of CENP-M as a pseudo GTPase scaffold required for HIKM tetramer assembly and downstream CENP-T/W recruitment, resolving where it acts in the kinetochore assembly hierarchy.","evidence":"Crystal structure, in vitro reconstitution, interface mutagenesis, Co-IP and live-cell loss-of-function imaging","pmids":["25006165"],"confidence":"High","gaps":["Why a GTPase fold is retained without nucleotide binding is unresolved","Dynamics of HIKM assembly across the cell cycle not characterized"]},{"year":2019,"claim":"Linked CENPM loss to suppressed proliferation and apoptosis in hepatocellular carcinoma and identified miR-1270 as a post-transcriptional regulator, framing CENPM as a tumor-promoting, regulated gene.","evidence":"siRNA knockdown, xenografts, flow cytometry, GSEA, and miRNA luciferase reporter assay","pmids":["31703591"],"confidence":"Medium","gaps":["p53/cell cycle pathway link rests on GSEA correlation rather than direct biochemistry","Whether phenotypes reflect kinetochore function or a separate role is unclear"]},{"year":2022,"claim":"Showed that the HPV E5 oncoprotein induces CENPM transcription through E2F1 binding at the CENPM promoter, providing a direct transcriptional mechanism for CENPM upregulation in cancer.","evidence":"ChIP for E2F1 promoter occupancy, siRNA and CRISPR knockdown, clonogenic survival assays in HNSCC","pmids":["35462155"],"confidence":"Medium","gaps":["Does not establish how CENPM levels alter radiosensitivity mechanistically beyond mitotic fraction","Generality of E2F1 control across tissues not tested here"]},{"year":2022,"claim":"Associated CENPM with mTOR signaling in lung adenocarcinoma and pancreatic cancer, proposing a non-kinetochore signaling axis driving proliferation and invasion.","evidence":"Gain/loss-of-function assays, Western blot for mTOR phosphorylation, mTOR inhibitor rescue, and xenografts","pmids":["35130633","32705259"],"confidence":"Low","gaps":["No biochemical mechanism linking CENPM to mTOR activation","Pathway placement rests on inhibitor rescue without upstream detail"]},{"year":2024,"claim":"Extended CENPM's cancer roles to glycolytic reprogramming and cGAS-STING/pyroptosis suppression, indicating influence over metabolism and innate immune signaling.","evidence":"shRNA/overexpression, glycolytic metabolite measurement with 2-DG rescue, and cGAS-STING inhibitor rescue with xenografts","pmids":["39707034","38693472"],"confidence":"Low","gaps":["No direct biochemical connection between CENPM and glycolytic enzymes or cGAS-STING components","Whether these effects are secondary to proliferation arrest is untested"]},{"year":2025,"claim":"Identified a physical interaction between CENPM and the immune checkpoint ligand FGL1 supporting ECM/collagen signaling and tumor invasion, the first direct non-kinetochore physical partner reported.","evidence":"Co-IP, DIA proteomics, immunofluorescence colocalization, and FGL1 rescue of migration/invasion in adrenocortical carcinoma","pmids":["39778025"],"confidence":"Medium","gaps":["Structural basis and stoichiometry of the CENPM-FGL1 interaction unknown","How a centromeric protein engages a secreted checkpoint ligand is unexplained"]},{"year":2025,"claim":"Implicated PI3K/AKT signaling in CENPM-driven glioma proliferation, adding another candidate signaling effector.","evidence":"shRNA knockdown, RNA-seq, Western blot for PI3K/AKT, cell cycle analysis, xenografts","pmids":["41317748"],"confidence":"Low","gaps":["Pathway link based on expression/Western data without rescue","No upstream mechanism connecting CENPM to PI3K/AKT activation"]},{"year":null,"claim":"How CENPM's defined centromeric scaffolding function mechanistically connects to the reported signaling, metabolic, and immune-modulatory roles in cancer remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No biochemical bridge between kinetochore function and mTOR/PI3K-AKT/glycolysis/cGAS-STING phenotypes","Unclear whether signaling effects are direct or downstream of mitotic/proliferative defects"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0]}],"complexes":["CENP-H/I/K/M (HIKM) tetramer","kinetochore"],"partners":["CENPI","CENPH","CENPK","FGL1","E2F1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NSP4","full_name":"Centromere protein M","aliases":["Interphase centromere complex protein 39","Proliferation-associated nuclear element protein 1"],"length_aa":180,"mass_kda":19.7,"function":"Component of the CENPA-NAC (nucleosome-associated) complex, a complex that plays a central role in assembly of kinetochore proteins, mitotic progression and chromosome segregation. The CENPA-NAC complex recruits the CENPA-CAD (nucleosome distal) complex and may be involved in incorporation of newly synthesized CENPA into centromeres","subcellular_location":"Nucleus; Cytoplasm; Chromosome, centromere, kinetochore","url":"https://www.uniprot.org/uniprotkb/Q9NSP4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CENPM","classification":"Common Essential","n_dependent_lines":1153,"n_total_lines":1208,"dependency_fraction":0.9544701986754967},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000100162","cell_line_id":"CID000926","localizations":[{"compartment":"nuclear_punctae","grade":3}],"interactors":[],"url":"https://opencell.sf.czbiohub.org/target/CID000926","total_profiled":1310},"omim":[{"mim_id":"611511","title":"MLF1-INTERACTING PROTEIN; MLF1IP","url":"https://www.omim.org/entry/611511"},{"mim_id":"611510","title":"CENTROMERIC PROTEIN T; CENPT","url":"https://www.omim.org/entry/611510"},{"mim_id":"611509","title":"CENTROMERIC PROTEIN N; CENPN","url":"https://www.omim.org/entry/611509"},{"mim_id":"611503","title":"CENTROMERIC PROTEIN L; CENPL","url":"https://www.omim.org/entry/611503"},{"mim_id":"610152","title":"CENTROMERIC PROTEIN M; CENPM","url":"https://www.omim.org/entry/610152"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":27.4},{"tissue":"lymphoid tissue","ntpm":23.4}],"url":"https://www.proteinatlas.org/search/CENPM"},"hgnc":{"alias_symbol":["Pane1","CENP-M","MGC861"],"prev_symbol":["C22orf18"]},"alphafold":{"accession":"Q9NSP4","domains":[{"cath_id":"3.40.50.300","chopping":"17-171","consensus_level":"high","plddt":92.5038,"start":17,"end":171}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NSP4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NSP4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NSP4-F1-predicted_aligned_error_v6.png","plddt_mean":89.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CENPM","jax_strain_url":"https://www.jax.org/strain/search?query=CENPM"},"sequence":{"accession":"Q9NSP4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NSP4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NSP4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NSP4"}},"corpus_meta":[{"pmid":"25006165","id":"PMC_25006165","title":"The pseudo GTPase CENP-M drives human kinetochore assembly.","date":"2014","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/25006165","citation_count":107,"is_preprint":false},{"pmid":"16391015","id":"PMC_16391015","title":"The PANE1 gene encodes a novel human minor histocompatibility antigen that is selectively expressed in B-lymphoid cells and B-CLL.","date":"2006","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/16391015","citation_count":83,"is_preprint":false},{"pmid":"31703591","id":"PMC_31703591","title":"Upregulation of CENPM promotes hepatocarcinogenesis through mutiple mechanisms.","date":"2019","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/31703591","citation_count":64,"is_preprint":false},{"pmid":"32663252","id":"PMC_32663252","title":"LncRNA HCG18 contributes to the progression of hepatocellular carcinoma via miR-214-3p/CENPM axis.","date":"2020","source":"Journal of biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32663252","citation_count":45,"is_preprint":false},{"pmid":"32705259","id":"PMC_32705259","title":"Upregulation of CENPM facilitates tumor metastasis via the mTOR/p70S6K signaling pathway in pancreatic cancer.","date":"2020","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/32705259","citation_count":21,"is_preprint":false},{"pmid":"38693472","id":"PMC_38693472","title":"Knockdown of CENPM activates cGAS-STING pathway to inhibit ovarian cancer by promoting pyroptosis.","date":"2024","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/38693472","citation_count":14,"is_preprint":false},{"pmid":"15183305","id":"PMC_15183305","title":"The proliferation associated nuclear element (PANE1) is conserved between mammals and fish and preferentially expressed in activated lymphoid cells.","date":"2004","source":"Gene expression patterns : GEP","url":"https://pubmed.ncbi.nlm.nih.gov/15183305","citation_count":13,"is_preprint":false},{"pmid":"34513693","id":"PMC_34513693","title":"ATF2-Induced Overexpression of lncRNA LINC00882, as a Novel Therapeutic Target, Accelerates Hepatocellular Carcinoma Progression via Sponging miR-214-3p to Upregulate CENPM.","date":"2021","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34513693","citation_count":11,"is_preprint":false},{"pmid":"35130633","id":"PMC_35130633","title":"Upregulation of CENPM facilitates lung adenocarcinoma progression via PI3K/AKT/mTOR signaling pathway.","date":"2022","source":"Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/35130633","citation_count":10,"is_preprint":false},{"pmid":"19711193","id":"PMC_19711193","title":"Molecular characterization and association analysis of porcine PANE1 gene.","date":"2009","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/19711193","citation_count":7,"is_preprint":false},{"pmid":"35462155","id":"PMC_35462155","title":"CENPM upregulation by E5 oncoprotein of human papillomavirus promotes radiosensitivity in head and neck squamous cell carcinoma.","date":"2022","source":"Oral oncology","url":"https://pubmed.ncbi.nlm.nih.gov/35462155","citation_count":6,"is_preprint":false},{"pmid":"39778025","id":"PMC_39778025","title":"Identification of CENPM as a key gene driving adrenocortical carcinoma metastasis via physical interaction with immune checkpoint ligand FGL1.","date":"2025","source":"Clinical and translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/39778025","citation_count":6,"is_preprint":false},{"pmid":"36635779","id":"PMC_36635779","title":"Upregulation of CENPM is associated with poor clinical outcome and suppression of immune profile in clear cell renal cell carcinoma.","date":"2023","source":"Hereditas","url":"https://pubmed.ncbi.nlm.nih.gov/36635779","citation_count":4,"is_preprint":false},{"pmid":"39707034","id":"PMC_39707034","title":"E2F1-driven CENPM expression promotes glycolytic reprogramming and tumorigenicity in glioblastoma.","date":"2024","source":"Cell biology and toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/39707034","citation_count":2,"is_preprint":false},{"pmid":"36036171","id":"PMC_36036171","title":"Minor histocompatibility antigens HA-8 and PANE1 in the TUNISIAN population.","date":"2022","source":"Molecular genetics & genomic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36036171","citation_count":1,"is_preprint":false},{"pmid":"41317748","id":"PMC_41317748","title":"Upregulation of CENPM facilitates glioma progression via PI3K/AKT signaling pathway.","date":"2025","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/41317748","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10078,"output_tokens":2637,"usd":0.034895,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9963,"output_tokens":2996,"usd":0.062358,"stage2_stop_reason":"end_turn"},"total_usd":0.097253,"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\": 2014,\n      \"finding\": \"CENP-M is structurally and evolutionarily related to small GTPases but is incapable of GTP-binding and conformational switching (pseudo GTPase). CENP-M is crucially required for the assembly and stability of the HIKM tetramer comprising CENP-I, CENP-H, and CENP-K. A point mutant disrupting the CENP-M/CENP-I interaction hampers kinetochore assembly and chromosome alignment, and prevents kinetochore recruitment of the CENP-T/W complex, placing CENP-M downstream of CENP-C in a single kinetochore assembly pathway.\",\n      \"method\": \"Crystal structure, in vitro biochemical reconstitution, Co-IP/pulldown, active-site/interface mutagenesis, live-cell imaging, cell biological loss-of-function assays\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure combined with biochemical reconstitution, mutagenesis, and cell biological validation in a single rigorous study\",\n      \"pmids\": [\"25006165\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"An alternative transcript of the PANE1/CENPM gene encodes a novel HLA-A*0301-restricted minor histocompatibility antigen (mHAg) selectively expressed in resting CD19+ B cells and B-CLL cells. The antigenic peptide is encoded within a unique exon, and a single nucleotide polymorphism replacing an arginine codon with a stop codon determines differential T-cell recognition. Activation of B-CLL cells via CD40L decreases expression of the mHAg-encoding transcript and reciprocally increases expression of transcripts lacking this exon.\",\n      \"method\": \"Sequencing of PANE1 alleles, mass spectrometry peptide identification, T-cell recognition assays, RT-PCR expression analysis\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (MS, sequencing, T-cell assay) in a single lab establishing isoform-specific antigen presentation\",\n      \"pmids\": [\"16391015\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CENPM knockdown in hepatocellular carcinoma cells inhibits cell proliferation in vitro and in vivo, suppresses migration and invasion, promotes apoptosis, and arrests the cell cycle. GSEA and functional validation linked CENPM activity to the p53 signaling pathway and cell cycle pathway. miR-1270 was identified as a negative post-transcriptional regulator of CENPM.\",\n      \"method\": \"siRNA knockdown, xenograft in vivo model, Western blot, flow cytometry, GSEA bioinformatics, miRNA luciferase reporter assay\",\n      \"journal\": \"Journal of experimental & clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined cellular phenotypes and pathway placement via GSEA plus miRNA reporter assay, single lab\",\n      \"pmids\": [\"31703591\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"CENPM knockdown in pancreatic cancer cells inhibits cell proliferation, alters the cell cycle, and limits cell migration and invasion via the mTOR/p70S6K signaling pathway.\",\n      \"method\": \"siRNA knockdown, cell proliferation assays, cell cycle analysis, migration/invasion assays, Western blot for mTOR/p70S6K pathway components\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement by Western blot without pharmacological rescue or upstream mechanistic detail\",\n      \"pmids\": [\"32705259\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CENPM promotes lung adenocarcinoma cell cycle progression, proliferation, migration and invasion by activating phosphorylation of mTOR (not by altering total mTOR protein levels). Inhibition of mTOR activity abrogates the promoting effects of CENPM overexpression.\",\n      \"method\": \"Gain/loss-of-function assays, Western blot for mTOR phosphorylation, mTOR inhibitor rescue experiment, xenograft model\",\n      \"journal\": \"Acta biochimica et biophysica Sinica\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement supported by inhibitor rescue but no upstream mechanistic detail of how CENPM activates mTOR\",\n      \"pmids\": [\"35130633\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"HPV E5 oncoprotein enhances CENPM expression through the transcription factor E2F1 binding to the CENPM promoter in head and neck squamous cell carcinoma. Inhibition of CENPM expression in HPV-positive HNSCC cells increases resistance to X-ray radiation (~59% under 2 Gy), associated with a reduced proportion of mitotic cells.\",\n      \"method\": \"ChIP assay for E2F1 binding at CENPM promoter, siRNA knockdown, CRISPR/Cas9 models, Western blot, qRT-PCR, clonogenic survival assay\",\n      \"journal\": \"Oral oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP directly demonstrates E2F1 binding to CENPM promoter; functional consequence validated by knockdown and CRISPR in single lab\",\n      \"pmids\": [\"35462155\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"E2F1 acts as a key transcriptional regulator of CENPM in glioblastoma. CENPM promotes GBM cell proliferation, invasion, and glycolytic reprogramming (glucose consumption, lactate production, ATP levels). The glycolytic inhibitor 2-DG reverses CENPM overexpression effects, indicating dependence on glycolytic pathways. In vivo, CENPM knockdown reduced tumor volume by ~60%.\",\n      \"method\": \"shRNA knockdown, vector-based overexpression, glycolytic metabolite measurement, 2-DG pharmacological rescue, xenograft mouse model\",\n      \"journal\": \"Cell biology and toxicology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement by rescue experiment but mechanistic link between CENPM and glycolysis is not biochemically characterized\",\n      \"pmids\": [\"39707034\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CENPM knockdown in ovarian cancer cells activates the cGAS-STING pathway and promotes pyroptosis, suppressing proliferation, migration and invasion. A cGAS-STING pathway inhibitor reversed the inhibitory effects of CENPM knockdown on viability, migration and invasion, placing CENPM upstream of cGAS-STING in ovarian cancer cells.\",\n      \"method\": \"siRNA knockdown, pharmacological pathway inhibitor rescue, ELISA for inflammatory factors, Western blot, xenograft model\",\n      \"journal\": \"BMC cancer\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement supported by inhibitor rescue but no direct biochemical link between CENPM and cGAS-STING activation\",\n      \"pmids\": [\"38693472\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CENPM physically interacts with immune checkpoint ligand FGL1 in adrenocortical carcinoma cells. CENPM knockdown downregulates FGL1, suppresses ECM-related collagen signaling, and inhibits ACC cell invasion and migration; overexpression of FGL1 rescues the migration and invasion defects caused by CENPM knockdown.\",\n      \"method\": \"Co-immunoprecipitation, DIA quantitative proteomics, immunofluorescence colocalization, Western blot, rescue overexpression experiment, xenograft NPG mice\",\n      \"journal\": \"Clinical and translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — physical interaction validated by Co-IP plus proteomics, functional consequence confirmed by rescue assay, single lab\",\n      \"pmids\": [\"39778025\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CENPM knockdown in glioma cells induces G0/G1 phase cell cycle arrest and inhibits proliferation. RNA-seq and Western blot analyses demonstrate that CENPM activates the PI3K/AKT signaling pathway in glioma cells; in vivo knockdown reduces tumor growth.\",\n      \"method\": \"shRNA knockdown, RNA-seq, Western blot for PI3K/AKT pathway, cell cycle flow cytometry, xenograft model\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement by Western blot and RNA-seq without upstream mechanistic detail or rescue\",\n      \"pmids\": [\"41317748\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CENPM (CENP-M) is a pseudo GTPase that is structurally related to small GTPases but cannot bind GTP; it is essential for assembling the CENP-H/I/K/M (HIKM) tetramer at the kinetochore–centromere interface downstream of CENP-C, thereby enabling CENP-T/W recruitment and faithful chromosome segregation during mitosis. In cancer contexts, CENPM expression is transcriptionally driven by E2F1 (and HPV E5 via E2F1), and its activity has been functionally linked to mTOR/p70S6K and PI3K/AKT signaling, glycolytic reprogramming, and physical interaction with the immune checkpoint ligand FGL1, though the biochemical mechanisms connecting CENPM to these non-kinetochore pathways remain incompletely characterized.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CENPM (CENP-M) is a centromere-associated protein essential for kinetochore assembly and faithful chromosome segregation during mitosis [#0]. Structurally and evolutionarily related to small GTPases, CENP-M is a pseudo GTPase that cannot bind GTP or undergo conformational switching; instead it functions as a scaffolding subunit required for the assembly and stability of the HIKM tetramer with CENP-I, CENP-H, and CENP-K [#0]. Disruption of the CENP-M/CENP-I interface impairs kinetochore assembly and chromosome alignment and blocks recruitment of the CENP-T/W complex, placing CENP-M in a single kinetochore assembly pathway downstream of CENP-C [#0]. An alternative transcript of the CENPM/PANE1 locus encodes an HLA-A*0301-restricted minor histocompatibility antigen selectively expressed in resting B cells and B-CLL cells, with a stop-codon polymorphism determining T-cell recognition [#1]. In cancer, CENPM expression is transcriptionally driven by E2F1, including induction by the HPV E5 oncoprotein via E2F1 binding to the CENPM promoter, where it supports mitotic progression and radiosensitivity [#5]. CENPM physically interacts with the immune checkpoint ligand FGL1 and sustains FGL1 levels and ECM-related collagen signaling to promote tumor cell invasion and migration [#8].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Established that the CENPM/PANE1 locus produces an alternative transcript encoding a tissue-restricted minor histocompatibility antigen, defining a non-mitotic, immunologically relevant output of the gene.\",\n      \"evidence\": \"Allele sequencing, mass spectrometry peptide identification, T-cell recognition and RT-PCR assays in B-CLL cells\",\n      \"pmids\": [\"16391015\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Does not connect the antigen-encoding isoform to the canonical centromeric function of CENP-M\",\n        \"Functional role of the alternative protein product beyond antigen presentation is unknown\"\n      ]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Defined the core molecular function of CENP-M as a pseudo GTPase scaffold required for HIKM tetramer assembly and downstream CENP-T/W recruitment, resolving where it acts in the kinetochore assembly hierarchy.\",\n      \"evidence\": \"Crystal structure, in vitro reconstitution, interface mutagenesis, Co-IP and live-cell loss-of-function imaging\",\n      \"pmids\": [\"25006165\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Why a GTPase fold is retained without nucleotide binding is unresolved\",\n        \"Dynamics of HIKM assembly across the cell cycle not characterized\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Linked CENPM loss to suppressed proliferation and apoptosis in hepatocellular carcinoma and identified miR-1270 as a post-transcriptional regulator, framing CENPM as a tumor-promoting, regulated gene.\",\n      \"evidence\": \"siRNA knockdown, xenografts, flow cytometry, GSEA, and miRNA luciferase reporter assay\",\n      \"pmids\": [\"31703591\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"p53/cell cycle pathway link rests on GSEA correlation rather than direct biochemistry\",\n        \"Whether phenotypes reflect kinetochore function or a separate role is unclear\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed that the HPV E5 oncoprotein induces CENPM transcription through E2F1 binding at the CENPM promoter, providing a direct transcriptional mechanism for CENPM upregulation in cancer.\",\n      \"evidence\": \"ChIP for E2F1 promoter occupancy, siRNA and CRISPR knockdown, clonogenic survival assays in HNSCC\",\n      \"pmids\": [\"35462155\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Does not establish how CENPM levels alter radiosensitivity mechanistically beyond mitotic fraction\",\n        \"Generality of E2F1 control across tissues not tested here\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Associated CENPM with mTOR signaling in lung adenocarcinoma and pancreatic cancer, proposing a non-kinetochore signaling axis driving proliferation and invasion.\",\n      \"evidence\": \"Gain/loss-of-function assays, Western blot for mTOR phosphorylation, mTOR inhibitor rescue, and xenografts\",\n      \"pmids\": [\"35130633\", \"32705259\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No biochemical mechanism linking CENPM to mTOR activation\",\n        \"Pathway placement rests on inhibitor rescue without upstream detail\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended CENPM's cancer roles to glycolytic reprogramming and cGAS-STING/pyroptosis suppression, indicating influence over metabolism and innate immune signaling.\",\n      \"evidence\": \"shRNA/overexpression, glycolytic metabolite measurement with 2-DG rescue, and cGAS-STING inhibitor rescue with xenografts\",\n      \"pmids\": [\"39707034\", \"38693472\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No direct biochemical connection between CENPM and glycolytic enzymes or cGAS-STING components\",\n        \"Whether these effects are secondary to proliferation arrest is untested\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified a physical interaction between CENPM and the immune checkpoint ligand FGL1 supporting ECM/collagen signaling and tumor invasion, the first direct non-kinetochore physical partner reported.\",\n      \"evidence\": \"Co-IP, DIA proteomics, immunofluorescence colocalization, and FGL1 rescue of migration/invasion in adrenocortical carcinoma\",\n      \"pmids\": [\"39778025\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Structural basis and stoichiometry of the CENPM-FGL1 interaction unknown\",\n        \"How a centromeric protein engages a secreted checkpoint ligand is unexplained\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Implicated PI3K/AKT signaling in CENPM-driven glioma proliferation, adding another candidate signaling effector.\",\n      \"evidence\": \"shRNA knockdown, RNA-seq, Western blot for PI3K/AKT, cell cycle analysis, xenografts\",\n      \"pmids\": [\"41317748\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Pathway link based on expression/Western data without rescue\",\n        \"No upstream mechanism connecting CENPM to PI3K/AKT activation\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How CENPM's defined centromeric scaffolding function mechanistically connects to the reported signaling, metabolic, and immune-modulatory roles in cancer remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No biochemical bridge between kinetochore function and mTOR/PI3K-AKT/glycolysis/cGAS-STING phenotypes\",\n        \"Unclear whether signaling effects are direct or downstream of mitotic/proliferative defects\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"complexes\": [\n      \"CENP-H/I/K/M (HIKM) tetramer\",\n      \"kinetochore\"\n    ],\n    \"partners\": [\n      \"CENPI\",\n      \"CENPH\",\n      \"CENPK\",\n      \"FGL1\",\n      \"E2F1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":6,"faith_pct":83.33333333333333}}