{"gene":"CNNM4","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2009,"finding":"CNNM4 (encoding a putative metal transporter) is the causative gene for Jalili syndrome (autosomal-recessive cone-rod dystrophy + amelogenesis imperfecta); expression confirmed in neural retina and ameloblasts of developing tooth, establishing a role for CNNM4-dependent metal transport in retinal function and tooth biomineralization.","method":"Positional-candidate gene approach; mutational analysis across seven families; expression analysis in retina and ameloblasts","journal":"American journal of human genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — independent replication in two simultaneous papers (PMID:19200525 and PMID:19200527) across multiple families with orthogonal genetic and expression methods","pmids":["19200525","19200527"],"is_preprint":false},{"year":2014,"finding":"CNNM4 is a membrane protein that stimulates Mg2+ efflux; it binds PRL (phosphatase of regenerating liver) family phosphatases, which suppress CNNM4-dependent Mg2+ efflux; regulation of intracellular Mg2+ by this PRL-CNNM4 axis is linked to energy metabolism and AMPK/mTOR signaling. In vivo, Cnnm4 deletion in ApcΔ14/+ mice promoted malignant progression of intestinal polyps to adenocarcinomas, demonstrating a tumor-suppressive role.","method":"Biochemical co-immunoprecipitation (CNNM4-PRL binding); intracellular Mg2+ measurements in cultured cells; Cnnm4 knockout mouse crossed with ApcΔ14/+ tumor model; mTOR inhibitor (rapamycin) rescue experiment","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP for binding, functional Mg2+ efflux assays, and in vivo genetic model with defined phenotypic readout in a single rigorous study","pmids":["25347473"],"is_preprint":false},{"year":2014,"finding":"CNNM4 localizes to the basolateral membrane of intestinal/polarized epithelia; this basolateral sorting requires interaction with clathrin adaptor protein complexes AP-1A and AP-1B via three conserved dileucine motifs in CNNM4. Simultaneous knockdown of μ1A and μ1B subunits abrogated basolateral localization.","method":"Immunofluorescence localization in MDCK cells; siRNA knockdown of AP-1A (μ1A) and AP-1B (μ1B) subunits; site-directed mutagenesis of dileucine motifs; co-immunoprecipitation of CNNM4 with μ1A and μ1B","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, mutagenesis, and KD with specific localization readout in a single study","pmids":["25449265"],"is_preprint":false},{"year":2016,"finding":"CNNM4 is required for Ca2+ influx during sperm capacitation; Cnnm4-deficient male mice are near-infertile due to abrogated sperm hyperactivation and perturbed Ca2+ influx. Forced Ca2+ entry into Cnnm4-deficient sperm normalized aberrant tyrosine phosphorylation, placing CNNM4 upstream of Ca2+ homeostasis in sperm capacitation.","method":"Cnnm4 germline and germ-cell-specific knockout mice; sperm motility analysis; Ca2+ imaging in live sperm; tyrosine phosphorylation western blot; forced Ca2+ entry rescue experiment","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean KO with defined cellular phenotype, Ca2+ imaging, and rescue experiment, all in a single rigorous study","pmids":["27006114"],"is_preprint":false},{"year":2019,"finding":"CNNM4 deficiency in colon epithelia impairs capsaicin-stimulated Ca2+ influx via TRPV1, promotes cell proliferation, and constitutively activates EGF receptor signaling; gefitinib (EGFR inhibitor) rescued the proliferation phenotype, functionally linking CNNM4-dependent Mg2+ transport to Ca2+ signaling and EGFR pathway regulation in colon epithelia.","method":"Cnnm4 knockout mice; organoid Ca2+ imaging with capsaicin stimulation; immunoblotting for EGFR signaling; gefitinib pharmacological rescue; comparison with Trpv1-deficient mice","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — KO mouse with defined cellular phenotype, Ca2+ imaging, epistasis with Trpv1 KO, and pharmacological rescue","pmids":["30670776"],"is_preprint":false},{"year":2005,"finding":"CNNM4 (as ACDP4) physically interacts with the intracellular metal ion chaperone COX11; co-expression of ACDP4 and COX11 in HEK293 cells enhanced toxicity to copper, manganese, and cobalt ions, suggesting functional coupling in essential metal ion delivery.","method":"Yeast two-hybrid screen of human fetal brain cDNA library; ectopic expression in HEK293 cells; metal ion toxicity assays","journal":"Molecular pain","confidence":"Medium","confidence_rationale":"Tier 3 / Weak — yeast two-hybrid plus cell-based toxicity assay; no reciprocal Co-IP or in vitro reconstitution; single lab","pmids":["15840172"],"is_preprint":false},{"year":2011,"finding":"The CBS-pair regulatory domain of human CNNM4 was successfully purified, overexpressed, and crystallized (orthorhombic space group C222), diffracting to 3.6 Å, with two molecules per asymmetric unit consistent with a CBS module dimer.","method":"Protein overexpression, purification, crystallization, and preliminary X-ray crystallographic analysis using synchrotron radiation","journal":"Acta crystallographica. Section F","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — crystal structure at preliminary resolution (3.6 Å), single study, no functional mutagenesis validation reported in abstract","pmids":["21393841"],"is_preprint":false},{"year":2018,"finding":"CNNM4 physically interacts with IQCB1 (a Leber congenital amaurosis gene product); a truncated CNNM4 protein (p.R605X) significantly increased apoptosis and enhanced the CNNM4-IQCB1 interaction, providing a functional link between Jalili syndrome and LCA pathways.","method":"Co-immunoprecipitation of CNNM4 and IQCB1; overexpression of truncated vs. full-length CNNM4 in cultured cells; apoptosis assay","journal":"Molecular genetics and genomics : MGG","confidence":"Medium","confidence_rationale":"Tier 3 / Weak — single Co-IP experiment, single lab, no reciprocal pull-down or in vitro reconstitution described","pmids":["29322253"],"is_preprint":false},{"year":2019,"finding":"CNNM4 CBS domain missense variants (p.Arg407Leu and p.Thr495Ile) were predicted by molecular dynamics simulations to disrupt ATP binding at the CBS domain, causing conformational shifts in the ATP binding site, suggesting ATP-dependent regulatory mechanism for CNNM4.","method":"Molecular dynamics simulations (60 ns); docking analysis of ATP to wild-type and mutant CBS domain models","journal":"Molecular genetics & genomic medicine","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational prediction only, no experimental validation of ATP binding or functional consequence","pmids":["31347285"],"is_preprint":false},{"year":2024,"finding":"CNNM4 CBS domain missense variants p.(Gly492Cys) and p.(Gly492Asp) exhibit reduced protein stability, increased mRNA decay rates, and significantly reduced Mg2+ extrusion activity compared to wild-type, despite normal subcellular localization; establishing that decreased mRNA/protein stability, not mislocalization, underlies the pathogenic mechanism.","method":"Mutant CNNM4 expression in cultured cells; Mg2+ efflux/extrusion functional assays; mRNA stability assays; subcellular localization imaging","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional Mg2+ transport assay combined with mRNA and protein stability measurements and localization analysis in a single study","pmids":["39580587"],"is_preprint":false},{"year":2024,"finding":"In thermogenic adipocytes, CNNM4 expression is induced by ADRB3-PKA-CREB signaling during cold exposure, mediating Mg2+ efflux; secreted Mg2+ then binds to the DFG motif of mTOR in macrophages to facilitate mTORC2 activation and M2 polarization.","method":"CNNM4 overexpression/knockdown in adipocytes; Mg2+ efflux measurements; signaling pathway inhibition (PKA/CREB); mTOR binding assay; macrophage polarization assays; adipose-specific Mg wire implantation in vivo","journal":"Advanced science","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — multiple functional assays in a single lab, mechanistic pathway established but Mg-mTOR binding claim relies on indirect evidence in the abstract","pmids":["39517124"],"is_preprint":false},{"year":2024,"finding":"CNNM4 overexpression in the liver causes Mg2+ homeostasis dysregulation linked to alcohol-associated liver disease progression; silencing Cnnm4 restores PCMT1 (protein isoaspartyl methyltransferase) activity and improves mitochondrial function, with the mechanism involving CNNM4-dependent regulation of S-adenosylmethionine levels required for PCMT1 function.","method":"GalNAc-siRNA silencing of Cnnm4 in mouse ALD model; PCMT1 activity assays; mitochondrial function measurements; S-adenosylmethionine level quantification; in vitro cell studies","journal":"Hepatology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA KD in vivo and in vitro with multiple functional readouts (enzymatic activity, metabolite levels, mitochondrial function) in a single study","pmids":["39641635"],"is_preprint":false},{"year":2025,"finding":"Multiple microRNAs regulate CNNM4 expression by directly binding the 3'UTR of CNNM4 mRNA; both up- and down-regulatory miRNAs were identified, and their modulation resulted in measurable changes in intracellular Mg2+ concentration in hepatocytes.","method":"High-throughput miRNA 3'UTR binding assay; miRNA overexpression in hepatocytes; intracellular Mg2+ concentration measurement","journal":"ACS chemical biology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct 3'UTR binding demonstrated with functional consequence (Mg2+ change) in cells; single lab, single study","pmids":["40862638"],"is_preprint":false},{"year":2026,"finding":"CNNM4 silencing in cholangiocarcinoma cells attenuates proliferation, chemoresistance, migration, invasion, cancer stem cell properties, and Warburg effect; proteomic analysis identified nuclear protein 1 (NUPR1) as an upstream regulator of CNNM4-induced ferroptosis, with iron chelation (deferiprone) reversing the anti-proliferative effect of CNNM4 silencing.","method":"siRNA/shRNA knockdown and GalNAc-conjugated siRNA in vivo; CCA cell line functional assays; chicken chorioallantoic membrane invasion model; proteomic analysis; pharmacological rescue with deferiprone and zinc protoporphyrin IX","journal":"Gut","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal functional assays in vitro and in vivo with proteomic pathway identification and pharmacological rescue, single lab","pmids":["40764063"],"is_preprint":false},{"year":2018,"finding":"CNNM4 and TRPM7 are co-expressed and co-localize in the odontoblastic process of rat odontoblasts, suggesting functional coupling of these two Mg2+ regulators in maintaining Mg2+ homeostasis in dentin formation.","method":"Single-cell RT-PCR of acutely dissociated rat odontoblasts; immunohistochemistry; immunocytochemistry co-localization analysis","journal":"Archives of oral biology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — co-localization without functional coupling experiment; single lab, no loss-of-function or interaction assay","pmids":["30278312"],"is_preprint":false}],"current_model":"CNNM4 is a plasma membrane Mg2+ efflux transporter that localizes to the basolateral membrane of polarized epithelia via AP-1A/AP-1B-dependent sorting through dileucine motifs; it is regulated post-transcriptionally by miRNAs and functionally inhibited by PRL phosphatase binding, which suppresses Mg2+ efflux to elevate intracellular Mg2+, thereby modulating AMPK/mTOR energy metabolism, EGFR and Ca2+ signaling (including TRPV1-dependent Ca2+ influx and sperm capacitation), and—through Mg2+ secretion—mTORC2 activation in neighboring macrophages; loss-of-function mutations in its CBS domain reduce Mg2+ transport and protein stability, causing Jalili syndrome (cone-rod dystrophy + amelogenesis imperfecta), while tissue-specific dysregulation of CNNM4-driven Mg2+ efflux contributes to cancer progression, alcohol-associated liver disease, and ferroptosis via NUPR1."},"narrative":{"mechanistic_narrative":"CNNM4 is a basolateral plasma-membrane Mg2+ efflux transporter of polarized epithelia that couples cellular magnesium homeostasis to growth, signaling, and metabolic programs [PMID:25347473, PMID:25449265]. Its basolateral targeting in epithelia depends on interaction with the clathrin adaptor complexes AP-1A and AP-1B through three conserved dileucine motifs [PMID:25449265]. CNNM4 stimulates Mg2+ extrusion and is functionally suppressed by binding to PRL-family phosphatases, an axis linking intracellular Mg2+ levels to AMPK/mTOR-dependent energy metabolism; loss of Cnnm4 drives malignant progression in an Apc-mutant intestinal tumor model, defining a tumor-suppressive role [PMID:25347473]. Through its control of Mg2+, CNNM4 governs downstream Ca2+ signaling: it is required for TRPV1-dependent Ca2+ influx and restraint of EGFR signaling in colon epithelia [PMID:30670776], and for Ca2+ influx during sperm capacitation and hyperactivation, placing it upstream of the Ca2+-dependent tyrosine phosphorylation that licenses fertilization [PMID:27006114]. CNNM4-mediated Mg2+ secretion also acts non-cell-autonomously, supporting mTORC2 activation and M2 polarization in neighboring macrophages during adipose thermogenesis [PMID:39517124], and its activity is tuned post-transcriptionally by 3'UTR-targeting microRNAs that alter intracellular Mg2+ [PMID:40862638]. Biallelic missense mutations in the CNNM4 CBS-pair regulatory domain cause Jalili syndrome (cone-rod dystrophy with amelogenesis imperfecta) [PMID:19200525, PMID:19200527], with pathogenic variants acting by reducing protein and mRNA stability and Mg2+ extrusion activity rather than by mislocalization [PMID:39580587]. Dysregulated CNNM4 activity further contributes to alcohol-associated liver disease via perturbed S-adenosylmethionine/PCMT1 function and mitochondrial dysfunction [PMID:39641635], and to cholangiocarcinoma progression through a NUPR1-linked ferroptosis axis [PMID:40764063].","teleology":[{"year":2009,"claim":"Established that CNNM4, a putative metal transporter, is the causative gene for Jalili syndrome, linking CNNM4-dependent metal transport to retinal function and tooth biomineralization.","evidence":"Positional-candidate gene mapping and mutational analysis across seven families with retinal/ameloblast expression analysis","pmids":["19200525","19200527"],"confidence":"High","gaps":["Did not define the transported ion or transport mechanism","Did not establish how mutations impair protein function"]},{"year":2014,"claim":"Defined CNNM4 as a Mg2+ efflux stimulator regulated by PRL phosphatase binding and tied this axis to AMPK/mTOR metabolism and tumor suppression, converting a 'putative transporter' into a defined regulatory node.","evidence":"Co-IP of CNNM4-PRL, intracellular Mg2+ assays, and Cnnm4 KO crossed into ApcΔ14/+ tumor model with rapamycin rescue","pmids":["25347473"],"confidence":"High","gaps":["Direct transport mechanism (channel vs exchanger) not resolved","Structural basis of PRL inhibition not defined"]},{"year":2014,"claim":"Explained how CNNM4 reaches its functional site by showing AP-1A/AP-1B and dileucine-motif-dependent basolateral sorting in polarized epithelia.","evidence":"MDCK immunofluorescence, μ1A/μ1B siRNA knockdown, dileucine mutagenesis, and Co-IP with μ subunits","pmids":["25449265"],"confidence":"High","gaps":["Whether the same sorting governs retinal/ameloblast localization untested","No structure of the AP-1/CNNM4 interface"]},{"year":2016,"claim":"Placed CNNM4 upstream of Ca2+ homeostasis in sperm capacitation, showing its loss causes near-infertility through abrogated Ca2+ influx and hyperactivation.","evidence":"Germline and germ-cell-specific Cnnm4 KO mice with sperm Ca2+ imaging, tyrosine phosphorylation blots, and forced Ca2+ entry rescue","pmids":["27006114"],"confidence":"High","gaps":["Molecular link between Mg2+ efflux and Ca2+ channel activation unresolved","Identity of the Ca2+ entry pathway in sperm not defined"]},{"year":2019,"claim":"Connected CNNM4-dependent Mg2+ transport to TRPV1 Ca2+ influx and EGFR signaling in colon epithelia, mechanistically tying its loss to proliferation.","evidence":"Cnnm4 KO mice and organoids, capsaicin Ca2+ imaging, Trpv1 KO epistasis, and gefitinib rescue","pmids":["30670776"],"confidence":"High","gaps":["How Mg2+ efflux is mechanistically coupled to TRPV1 unclear","Whether EGFR activation is direct or downstream of Ca2+ not separated"]},{"year":2024,"claim":"Showed Jalili-causing CBS-domain missense variants act by reducing mRNA/protein stability and Mg2+ extrusion rather than by mislocalization, resolving the molecular basis of pathogenicity.","evidence":"Mutant CNNM4 expression with Mg2+ efflux assays, mRNA stability assays, and localization imaging","pmids":["39580587"],"confidence":"Medium","gaps":["In vivo consequences of destabilized protein not modeled","Does not explain tissue specificity of retinal/dental phenotypes"]},{"year":2024,"claim":"Revealed a non-cell-autonomous role: cold-induced CNNM4 in thermogenic adipocytes secretes Mg2+ that promotes mTORC2 activation and M2 macrophage polarization.","evidence":"Adipocyte CNNM4 overexpression/knockdown, Mg2+ efflux and mTOR binding assays, macrophage polarization, and adipose Mg-wire implantation in vivo","pmids":["39517124"],"confidence":"Medium","gaps":["Direct Mg2+-mTOR DFG-motif binding rests on indirect evidence","Physiological range of secreted Mg2+ not quantified"]},{"year":2024,"claim":"Implicated CNNM4 in alcohol-associated liver disease through Mg2+-dependent control of S-adenosylmethionine/PCMT1 activity and mitochondrial function.","evidence":"GalNAc-siRNA Cnnm4 silencing in mouse ALD model with PCMT1 activity, SAM quantification, and mitochondrial assays","pmids":["39641635"],"confidence":"Medium","gaps":["Mechanistic link from Mg2+ to SAM levels not fully defined","Causality vs correlation of mitochondrial improvement not isolated"]},{"year":2025,"claim":"Established post-transcriptional control of CNNM4 by multiple 3'UTR-binding microRNAs that tune intracellular Mg2+ in hepatocytes.","evidence":"High-throughput miRNA 3'UTR binding assay with miRNA overexpression and intracellular Mg2+ measurement","pmids":["40862638"],"confidence":"Medium","gaps":["Physiological contexts driving these miRNAs unknown","In vivo relevance not tested"]},{"year":2026,"claim":"Linked CNNM4 to cholangiocarcinoma aggressiveness and identified a NUPR1-associated ferroptosis axis as the mechanistic pathway.","evidence":"siRNA/shRNA and GalNAc-siRNA knockdown in vitro and in vivo, CAM invasion model, proteomics, and deferiprone/ZnPP rescue","pmids":["40764063"],"confidence":"Medium","gaps":["Direction of CNNM4-NUPR1 regulation not fully mechanistically resolved","Link between Mg2+ transport and iron/ferroptosis unclear"]},{"year":null,"claim":"The structural mechanism by which CNNM4 transports Mg2+, how its CBS-pair/ATP-binding module gates transport, and how Mg2+ efflux is mechanistically coupled to downstream Ca2+, EGFR, mTOR, and ferroptosis pathways remain unresolved.","evidence":"No reconstituted transport or full-length structure in the timeline; CBS-domain crystallization and ATP-docking remain preliminary","pmids":[],"confidence":"Low","gaps":["No high-resolution structure of full-length transporter","ATP regulation of CBS domain unvalidated experimentally","Transport stoichiometry and electrogenicity unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0005215","term_label":"transporter activity","supporting_discovery_ids":[1,2,9,10]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,2]}],"pathway":[{"term_id":"R-HSA-382551","term_label":"Transport of small molecules","supporting_discovery_ids":[1,9,10]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[4,10]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[0,11,13]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[13]}],"complexes":[],"partners":["PRL PHOSPHATASES","AP-1A (Μ1A)","AP-1B (Μ1B)","COX11","IQCB1","TRPV1","TRPM7","PCMT1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q6P4Q7","full_name":"Metal transporter CNNM4","aliases":["Ancient conserved domain-containing protein 4","Cyclin-M4"],"length_aa":775,"mass_kda":86.6,"function":"Probable metal transporter. The interaction with the metal ion chaperone COX11 suggests that it may play a role in sensory neuron functions (By similarity). May play a role in biomineralization and retinal function","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q6P4Q7/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CNNM4","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CNNM4","total_profiled":1310},"omim":[{"mim_id":"607805","title":"CYCLIN M4; CNNM4","url":"https://www.omim.org/entry/607805"},{"mim_id":"607804","title":"CYCLIN M3; CNNM3","url":"https://www.omim.org/entry/607804"},{"mim_id":"217080","title":"JALILI SYNDROME","url":"https://www.omim.org/entry/217080"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Uncertain","locations":[{"location":"Actin filaments","reliability":"Uncertain"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"retina","ntpm":36.0}],"url":"https://www.proteinatlas.org/search/CNNM4"},"hgnc":{"alias_symbol":["SLC70A4","KIAA1592"],"prev_symbol":["ACDP4"]},"alphafold":{"accession":"Q6P4Q7","domains":[{"cath_id":"2.60.40,2.60.40","chopping":"46-174","consensus_level":"high","plddt":81.1621,"start":46,"end":174},{"cath_id":"-","chopping":"183-329","consensus_level":"high","plddt":88.6172,"start":183,"end":329},{"cath_id":"3.10.580.10","chopping":"370-518","consensus_level":"high","plddt":90.0452,"start":370,"end":518},{"cath_id":"2.60.120.10","chopping":"544-647_695-727","consensus_level":"high","plddt":89.1387,"start":544,"end":727}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6P4Q7","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q6P4Q7-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q6P4Q7-F1-predicted_aligned_error_v6.png","plddt_mean":76.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CNNM4","jax_strain_url":"https://www.jax.org/strain/search?query=CNNM4"},"sequence":{"accession":"Q6P4Q7","fasta_url":"https://rest.uniprot.org/uniprotkb/Q6P4Q7.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q6P4Q7/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6P4Q7"}},"corpus_meta":[{"pmid":"19200525","id":"PMC_19200525","title":"Mutations in CNNM4 cause Jalili syndrome, consisting of autosomal-recessive cone-rod dystrophy and amelogenesis imperfecta.","date":"2009","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/19200525","citation_count":130,"is_preprint":false},{"pmid":"25347473","id":"PMC_25347473","title":"Membrane protein CNNM4-dependent Mg2+ efflux suppresses tumor progression.","date":"2014","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/25347473","citation_count":98,"is_preprint":false},{"pmid":"19200527","id":"PMC_19200527","title":"Mutations in CNNM4 cause recessive cone-rod dystrophy with amelogenesis imperfecta.","date":"2009","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/19200527","citation_count":81,"is_preprint":false},{"pmid":"27006114","id":"PMC_27006114","title":"The Mg2+ transporter CNNM4 regulates sperm Ca2+ homeostasis and is essential for reproduction.","date":"2016","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/27006114","citation_count":34,"is_preprint":false},{"pmid":"24194943","id":"PMC_24194943","title":"Dental phenotype in Jalili syndrome due to a c.1312 dupC homozygous mutation in the CNNM4 gene.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24194943","citation_count":30,"is_preprint":false},{"pmid":"15840172","id":"PMC_15840172","title":"Physical interaction and functional coupling between ACDP4 and the intracellular ion chaperone COX11, an implication of the role of ACDP4 in essential metal ion transport and homeostasis.","date":"2005","source":"Molecular pain","url":"https://pubmed.ncbi.nlm.nih.gov/15840172","citation_count":23,"is_preprint":false},{"pmid":"30670776","id":"PMC_30670776","title":"Cnnm4 deficiency suppresses Ca2+ signaling and promotes cell proliferation in the colon epithelia.","date":"2019","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/30670776","citation_count":19,"is_preprint":false},{"pmid":"29322253","id":"PMC_29322253","title":"Identification of a mutation in CNNM4 by whole exome sequencing in an Amish family and functional link between CNNM4 and IQCB1.","date":"2018","source":"Molecular genetics and genomics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/29322253","citation_count":15,"is_preprint":false},{"pmid":"27070327","id":"PMC_27070327","title":"A new familial case of Jalili syndrome caused by a novel mutation in CNNM4.","date":"2016","source":"Ophthalmic genetics","url":"https://pubmed.ncbi.nlm.nih.gov/27070327","citation_count":15,"is_preprint":false},{"pmid":"30278312","id":"PMC_30278312","title":"Molecular expression of Mg2+ regulator TRPM7 and CNNM4 in rat odontoblasts.","date":"2018","source":"Archives of oral biology","url":"https://pubmed.ncbi.nlm.nih.gov/30278312","citation_count":14,"is_preprint":false},{"pmid":"31347285","id":"PMC_31347285","title":"A novel pathogenic missense variant in CNNM4 underlying Jalili syndrome: Insights from molecular dynamics simulations.","date":"2019","source":"Molecular genetics & genomic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31347285","citation_count":13,"is_preprint":false},{"pmid":"29421602","id":"PMC_29421602","title":"Report of two unrelated families with Jalili syndrome and a novel nonsense heterozygous mutation in CNNM4 gene.","date":"2018","source":"European journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29421602","citation_count":13,"is_preprint":false},{"pmid":"21393841","id":"PMC_21393841","title":"Purification, crystallization and preliminary crystallographic analysis of the CBS pair of the human metal transporter CNNM4.","date":"2011","source":"Acta crystallographica. Section F, Structural biology and crystallization communications","url":"https://pubmed.ncbi.nlm.nih.gov/21393841","citation_count":11,"is_preprint":false},{"pmid":"25449265","id":"PMC_25449265","title":"Basolateral sorting of the Mg²⁺ transporter CNNM4 requires interaction with AP-1A and AP-1B.","date":"2014","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/25449265","citation_count":11,"is_preprint":false},{"pmid":"39641635","id":"PMC_39641635","title":"Modulatory effects of CNNM4 on protein- l -isoaspartyl- O -methyltransferase repair function during alcohol-induced hepatic damage.","date":"2024","source":"Hepatology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/39641635","citation_count":9,"is_preprint":false},{"pmid":"28246031","id":"PMC_28246031","title":"Novel splice site mutation in CNNM4 gene in a family with Jalili syndrome.","date":"2017","source":"European journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/28246031","citation_count":9,"is_preprint":false},{"pmid":"32022389","id":"PMC_32022389","title":"Expanding the genotypic spectrum of Jalili syndrome: Novel CNNM4 variants and uniparental isodisomy in a north American patient cohort.","date":"2020","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/32022389","citation_count":9,"is_preprint":false},{"pmid":"39517124","id":"PMC_39517124","title":"Thermogenic Adipocytes Promote M2 Macrophage Polarization through CNNM4-Mediated Mg Secretion.","date":"2024","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/39517124","citation_count":6,"is_preprint":false},{"pmid":"34875963","id":"PMC_34875963","title":"Cone pathway dysfunction in Jalili syndrome due to a novel familial variant of CNNM4 revealed by pupillometry and electrophysiologic investigations.","date":"2021","source":"Ophthalmic genetics","url":"https://pubmed.ncbi.nlm.nih.gov/34875963","citation_count":6,"is_preprint":false},{"pmid":"35150469","id":"PMC_35150469","title":"Novel homozygous nonsynonymous variant of CNNM4 gene in a Chinese family with Jalili syndrome.","date":"2022","source":"Molecular genetics & genomic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/35150469","citation_count":6,"is_preprint":false},{"pmid":"40764063","id":"PMC_40764063","title":"Role of CNNM4 in the progression of cholangiocarcinoma: implications for ferroptosis and therapeutic potential.","date":"2026","source":"Gut","url":"https://pubmed.ncbi.nlm.nih.gov/40764063","citation_count":3,"is_preprint":false},{"pmid":"39580587","id":"PMC_39580587","title":"Functional and pathogenic insights into CNNM4 variants in Jalili syndrome.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39580587","citation_count":1,"is_preprint":false},{"pmid":"40232358","id":"PMC_40232358","title":"A novel mutation in CNNM4 is associated with a case of Jalili syndrome in Egypt.","date":"2025","source":"Documenta ophthalmologica. Advances in ophthalmology","url":"https://pubmed.ncbi.nlm.nih.gov/40232358","citation_count":1,"is_preprint":false},{"pmid":"40862638","id":"PMC_40862638","title":"microRNAs Regulate Cellular Magnesium by Tuning Expression of the Plasma Membrane Protein CNNM4.","date":"2025","source":"ACS chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/40862638","citation_count":0,"is_preprint":false},{"pmid":"41280631","id":"PMC_41280631","title":"Novel mutation in CNNM4 gene in a Chinese family with Jalili syndrome and literature review.","date":"2025","source":"International journal of ophthalmology","url":"https://pubmed.ncbi.nlm.nih.gov/41280631","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14132,"output_tokens":4046,"usd":0.051543,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11919,"output_tokens":3823,"usd":0.077585,"stage2_stop_reason":"end_turn"},"total_usd":0.129128,"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\": 2009,\n      \"finding\": \"CNNM4 (encoding a putative metal transporter) is the causative gene for Jalili syndrome (autosomal-recessive cone-rod dystrophy + amelogenesis imperfecta); expression confirmed in neural retina and ameloblasts of developing tooth, establishing a role for CNNM4-dependent metal transport in retinal function and tooth biomineralization.\",\n      \"method\": \"Positional-candidate gene approach; mutational analysis across seven families; expression analysis in retina and ameloblasts\",\n      \"journal\": \"American journal of human genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — independent replication in two simultaneous papers (PMID:19200525 and PMID:19200527) across multiple families with orthogonal genetic and expression methods\",\n      \"pmids\": [\"19200525\", \"19200527\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CNNM4 is a membrane protein that stimulates Mg2+ efflux; it binds PRL (phosphatase of regenerating liver) family phosphatases, which suppress CNNM4-dependent Mg2+ efflux; regulation of intracellular Mg2+ by this PRL-CNNM4 axis is linked to energy metabolism and AMPK/mTOR signaling. In vivo, Cnnm4 deletion in ApcΔ14/+ mice promoted malignant progression of intestinal polyps to adenocarcinomas, demonstrating a tumor-suppressive role.\",\n      \"method\": \"Biochemical co-immunoprecipitation (CNNM4-PRL binding); intracellular Mg2+ measurements in cultured cells; Cnnm4 knockout mouse crossed with ApcΔ14/+ tumor model; mTOR inhibitor (rapamycin) rescue experiment\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP for binding, functional Mg2+ efflux assays, and in vivo genetic model with defined phenotypic readout in a single rigorous study\",\n      \"pmids\": [\"25347473\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CNNM4 localizes to the basolateral membrane of intestinal/polarized epithelia; this basolateral sorting requires interaction with clathrin adaptor protein complexes AP-1A and AP-1B via three conserved dileucine motifs in CNNM4. Simultaneous knockdown of μ1A and μ1B subunits abrogated basolateral localization.\",\n      \"method\": \"Immunofluorescence localization in MDCK cells; siRNA knockdown of AP-1A (μ1A) and AP-1B (μ1B) subunits; site-directed mutagenesis of dileucine motifs; co-immunoprecipitation of CNNM4 with μ1A and μ1B\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, mutagenesis, and KD with specific localization readout in a single study\",\n      \"pmids\": [\"25449265\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CNNM4 is required for Ca2+ influx during sperm capacitation; Cnnm4-deficient male mice are near-infertile due to abrogated sperm hyperactivation and perturbed Ca2+ influx. Forced Ca2+ entry into Cnnm4-deficient sperm normalized aberrant tyrosine phosphorylation, placing CNNM4 upstream of Ca2+ homeostasis in sperm capacitation.\",\n      \"method\": \"Cnnm4 germline and germ-cell-specific knockout mice; sperm motility analysis; Ca2+ imaging in live sperm; tyrosine phosphorylation western blot; forced Ca2+ entry rescue experiment\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO with defined cellular phenotype, Ca2+ imaging, and rescue experiment, all in a single rigorous study\",\n      \"pmids\": [\"27006114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CNNM4 deficiency in colon epithelia impairs capsaicin-stimulated Ca2+ influx via TRPV1, promotes cell proliferation, and constitutively activates EGF receptor signaling; gefitinib (EGFR inhibitor) rescued the proliferation phenotype, functionally linking CNNM4-dependent Mg2+ transport to Ca2+ signaling and EGFR pathway regulation in colon epithelia.\",\n      \"method\": \"Cnnm4 knockout mice; organoid Ca2+ imaging with capsaicin stimulation; immunoblotting for EGFR signaling; gefitinib pharmacological rescue; comparison with Trpv1-deficient mice\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse with defined cellular phenotype, Ca2+ imaging, epistasis with Trpv1 KO, and pharmacological rescue\",\n      \"pmids\": [\"30670776\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"CNNM4 (as ACDP4) physically interacts with the intracellular metal ion chaperone COX11; co-expression of ACDP4 and COX11 in HEK293 cells enhanced toxicity to copper, manganese, and cobalt ions, suggesting functional coupling in essential metal ion delivery.\",\n      \"method\": \"Yeast two-hybrid screen of human fetal brain cDNA library; ectopic expression in HEK293 cells; metal ion toxicity assays\",\n      \"journal\": \"Molecular pain\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Weak — yeast two-hybrid plus cell-based toxicity assay; no reciprocal Co-IP or in vitro reconstitution; single lab\",\n      \"pmids\": [\"15840172\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"The CBS-pair regulatory domain of human CNNM4 was successfully purified, overexpressed, and crystallized (orthorhombic space group C222), diffracting to 3.6 Å, with two molecules per asymmetric unit consistent with a CBS module dimer.\",\n      \"method\": \"Protein overexpression, purification, crystallization, and preliminary X-ray crystallographic analysis using synchrotron radiation\",\n      \"journal\": \"Acta crystallographica. Section F\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — crystal structure at preliminary resolution (3.6 Å), single study, no functional mutagenesis validation reported in abstract\",\n      \"pmids\": [\"21393841\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CNNM4 physically interacts with IQCB1 (a Leber congenital amaurosis gene product); a truncated CNNM4 protein (p.R605X) significantly increased apoptosis and enhanced the CNNM4-IQCB1 interaction, providing a functional link between Jalili syndrome and LCA pathways.\",\n      \"method\": \"Co-immunoprecipitation of CNNM4 and IQCB1; overexpression of truncated vs. full-length CNNM4 in cultured cells; apoptosis assay\",\n      \"journal\": \"Molecular genetics and genomics : MGG\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP experiment, single lab, no reciprocal pull-down or in vitro reconstitution described\",\n      \"pmids\": [\"29322253\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CNNM4 CBS domain missense variants (p.Arg407Leu and p.Thr495Ile) were predicted by molecular dynamics simulations to disrupt ATP binding at the CBS domain, causing conformational shifts in the ATP binding site, suggesting ATP-dependent regulatory mechanism for CNNM4.\",\n      \"method\": \"Molecular dynamics simulations (60 ns); docking analysis of ATP to wild-type and mutant CBS domain models\",\n      \"journal\": \"Molecular genetics & genomic medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational prediction only, no experimental validation of ATP binding or functional consequence\",\n      \"pmids\": [\"31347285\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CNNM4 CBS domain missense variants p.(Gly492Cys) and p.(Gly492Asp) exhibit reduced protein stability, increased mRNA decay rates, and significantly reduced Mg2+ extrusion activity compared to wild-type, despite normal subcellular localization; establishing that decreased mRNA/protein stability, not mislocalization, underlies the pathogenic mechanism.\",\n      \"method\": \"Mutant CNNM4 expression in cultured cells; Mg2+ efflux/extrusion functional assays; mRNA stability assays; subcellular localization imaging\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional Mg2+ transport assay combined with mRNA and protein stability measurements and localization analysis in a single study\",\n      \"pmids\": [\"39580587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In thermogenic adipocytes, CNNM4 expression is induced by ADRB3-PKA-CREB signaling during cold exposure, mediating Mg2+ efflux; secreted Mg2+ then binds to the DFG motif of mTOR in macrophages to facilitate mTORC2 activation and M2 polarization.\",\n      \"method\": \"CNNM4 overexpression/knockdown in adipocytes; Mg2+ efflux measurements; signaling pathway inhibition (PKA/CREB); mTOR binding assay; macrophage polarization assays; adipose-specific Mg wire implantation in vivo\",\n      \"journal\": \"Advanced science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — multiple functional assays in a single lab, mechanistic pathway established but Mg-mTOR binding claim relies on indirect evidence in the abstract\",\n      \"pmids\": [\"39517124\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CNNM4 overexpression in the liver causes Mg2+ homeostasis dysregulation linked to alcohol-associated liver disease progression; silencing Cnnm4 restores PCMT1 (protein isoaspartyl methyltransferase) activity and improves mitochondrial function, with the mechanism involving CNNM4-dependent regulation of S-adenosylmethionine levels required for PCMT1 function.\",\n      \"method\": \"GalNAc-siRNA silencing of Cnnm4 in mouse ALD model; PCMT1 activity assays; mitochondrial function measurements; S-adenosylmethionine level quantification; in vitro cell studies\",\n      \"journal\": \"Hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA KD in vivo and in vitro with multiple functional readouts (enzymatic activity, metabolite levels, mitochondrial function) in a single study\",\n      \"pmids\": [\"39641635\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Multiple microRNAs regulate CNNM4 expression by directly binding the 3'UTR of CNNM4 mRNA; both up- and down-regulatory miRNAs were identified, and their modulation resulted in measurable changes in intracellular Mg2+ concentration in hepatocytes.\",\n      \"method\": \"High-throughput miRNA 3'UTR binding assay; miRNA overexpression in hepatocytes; intracellular Mg2+ concentration measurement\",\n      \"journal\": \"ACS chemical biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct 3'UTR binding demonstrated with functional consequence (Mg2+ change) in cells; single lab, single study\",\n      \"pmids\": [\"40862638\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CNNM4 silencing in cholangiocarcinoma cells attenuates proliferation, chemoresistance, migration, invasion, cancer stem cell properties, and Warburg effect; proteomic analysis identified nuclear protein 1 (NUPR1) as an upstream regulator of CNNM4-induced ferroptosis, with iron chelation (deferiprone) reversing the anti-proliferative effect of CNNM4 silencing.\",\n      \"method\": \"siRNA/shRNA knockdown and GalNAc-conjugated siRNA in vivo; CCA cell line functional assays; chicken chorioallantoic membrane invasion model; proteomic analysis; pharmacological rescue with deferiprone and zinc protoporphyrin IX\",\n      \"journal\": \"Gut\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal functional assays in vitro and in vivo with proteomic pathway identification and pharmacological rescue, single lab\",\n      \"pmids\": [\"40764063\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CNNM4 and TRPM7 are co-expressed and co-localize in the odontoblastic process of rat odontoblasts, suggesting functional coupling of these two Mg2+ regulators in maintaining Mg2+ homeostasis in dentin formation.\",\n      \"method\": \"Single-cell RT-PCR of acutely dissociated rat odontoblasts; immunohistochemistry; immunocytochemistry co-localization analysis\",\n      \"journal\": \"Archives of oral biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — co-localization without functional coupling experiment; single lab, no loss-of-function or interaction assay\",\n      \"pmids\": [\"30278312\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CNNM4 is a plasma membrane Mg2+ efflux transporter that localizes to the basolateral membrane of polarized epithelia via AP-1A/AP-1B-dependent sorting through dileucine motifs; it is regulated post-transcriptionally by miRNAs and functionally inhibited by PRL phosphatase binding, which suppresses Mg2+ efflux to elevate intracellular Mg2+, thereby modulating AMPK/mTOR energy metabolism, EGFR and Ca2+ signaling (including TRPV1-dependent Ca2+ influx and sperm capacitation), and—through Mg2+ secretion—mTORC2 activation in neighboring macrophages; loss-of-function mutations in its CBS domain reduce Mg2+ transport and protein stability, causing Jalili syndrome (cone-rod dystrophy + amelogenesis imperfecta), while tissue-specific dysregulation of CNNM4-driven Mg2+ efflux contributes to cancer progression, alcohol-associated liver disease, and ferroptosis via NUPR1.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CNNM4 is a basolateral plasma-membrane Mg2+ efflux transporter of polarized epithelia that couples cellular magnesium homeostasis to growth, signaling, and metabolic programs [#1, #2]. Its basolateral targeting in epithelia depends on interaction with the clathrin adaptor complexes AP-1A and AP-1B through three conserved dileucine motifs [#2]. CNNM4 stimulates Mg2+ extrusion and is functionally suppressed by binding to PRL-family phosphatases, an axis linking intracellular Mg2+ levels to AMPK/mTOR-dependent energy metabolism; loss of Cnnm4 drives malignant progression in an Apc-mutant intestinal tumor model, defining a tumor-suppressive role [#1]. Through its control of Mg2+, CNNM4 governs downstream Ca2+ signaling: it is required for TRPV1-dependent Ca2+ influx and restraint of EGFR signaling in colon epithelia [#4], and for Ca2+ influx during sperm capacitation and hyperactivation, placing it upstream of the Ca2+-dependent tyrosine phosphorylation that licenses fertilization [#3]. CNNM4-mediated Mg2+ secretion also acts non-cell-autonomously, supporting mTORC2 activation and M2 polarization in neighboring macrophages during adipose thermogenesis [#10], and its activity is tuned post-transcriptionally by 3'UTR-targeting microRNAs that alter intracellular Mg2+ [#12]. Biallelic missense mutations in the CNNM4 CBS-pair regulatory domain cause Jalili syndrome (cone-rod dystrophy with amelogenesis imperfecta) [#0], with pathogenic variants acting by reducing protein and mRNA stability and Mg2+ extrusion activity rather than by mislocalization [#9]. Dysregulated CNNM4 activity further contributes to alcohol-associated liver disease via perturbed S-adenosylmethionine/PCMT1 function and mitochondrial dysfunction [#11], and to cholangiocarcinoma progression through a NUPR1-linked ferroptosis axis [#13].\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Established that CNNM4, a putative metal transporter, is the causative gene for Jalili syndrome, linking CNNM4-dependent metal transport to retinal function and tooth biomineralization.\",\n      \"evidence\": \"Positional-candidate gene mapping and mutational analysis across seven families with retinal/ameloblast expression analysis\",\n      \"pmids\": [\"19200525\", \"19200527\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define the transported ion or transport mechanism\", \"Did not establish how mutations impair protein function\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Defined CNNM4 as a Mg2+ efflux stimulator regulated by PRL phosphatase binding and tied this axis to AMPK/mTOR metabolism and tumor suppression, converting a 'putative transporter' into a defined regulatory node.\",\n      \"evidence\": \"Co-IP of CNNM4-PRL, intracellular Mg2+ assays, and Cnnm4 KO crossed into Apc\\u039414/+ tumor model with rapamycin rescue\",\n      \"pmids\": [\"25347473\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct transport mechanism (channel vs exchanger) not resolved\", \"Structural basis of PRL inhibition not defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Explained how CNNM4 reaches its functional site by showing AP-1A/AP-1B and dileucine-motif-dependent basolateral sorting in polarized epithelia.\",\n      \"evidence\": \"MDCK immunofluorescence, \\u03bc1A/\\u03bc1B siRNA knockdown, dileucine mutagenesis, and Co-IP with \\u03bc subunits\",\n      \"pmids\": [\"25449265\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the same sorting governs retinal/ameloblast localization untested\", \"No structure of the AP-1/CNNM4 interface\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Placed CNNM4 upstream of Ca2+ homeostasis in sperm capacitation, showing its loss causes near-infertility through abrogated Ca2+ influx and hyperactivation.\",\n      \"evidence\": \"Germline and germ-cell-specific Cnnm4 KO mice with sperm Ca2+ imaging, tyrosine phosphorylation blots, and forced Ca2+ entry rescue\",\n      \"pmids\": [\"27006114\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular link between Mg2+ efflux and Ca2+ channel activation unresolved\", \"Identity of the Ca2+ entry pathway in sperm not defined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Connected CNNM4-dependent Mg2+ transport to TRPV1 Ca2+ influx and EGFR signaling in colon epithelia, mechanistically tying its loss to proliferation.\",\n      \"evidence\": \"Cnnm4 KO mice and organoids, capsaicin Ca2+ imaging, Trpv1 KO epistasis, and gefitinib rescue\",\n      \"pmids\": [\"30670776\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How Mg2+ efflux is mechanistically coupled to TRPV1 unclear\", \"Whether EGFR activation is direct or downstream of Ca2+ not separated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showed Jalili-causing CBS-domain missense variants act by reducing mRNA/protein stability and Mg2+ extrusion rather than by mislocalization, resolving the molecular basis of pathogenicity.\",\n      \"evidence\": \"Mutant CNNM4 expression with Mg2+ efflux assays, mRNA stability assays, and localization imaging\",\n      \"pmids\": [\"39580587\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo consequences of destabilized protein not modeled\", \"Does not explain tissue specificity of retinal/dental phenotypes\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed a non-cell-autonomous role: cold-induced CNNM4 in thermogenic adipocytes secretes Mg2+ that promotes mTORC2 activation and M2 macrophage polarization.\",\n      \"evidence\": \"Adipocyte CNNM4 overexpression/knockdown, Mg2+ efflux and mTOR binding assays, macrophage polarization, and adipose Mg-wire implantation in vivo\",\n      \"pmids\": [\"39517124\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct Mg2+-mTOR DFG-motif binding rests on indirect evidence\", \"Physiological range of secreted Mg2+ not quantified\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Implicated CNNM4 in alcohol-associated liver disease through Mg2+-dependent control of S-adenosylmethionine/PCMT1 activity and mitochondrial function.\",\n      \"evidence\": \"GalNAc-siRNA Cnnm4 silencing in mouse ALD model with PCMT1 activity, SAM quantification, and mitochondrial assays\",\n      \"pmids\": [\"39641635\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanistic link from Mg2+ to SAM levels not fully defined\", \"Causality vs correlation of mitochondrial improvement not isolated\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Established post-transcriptional control of CNNM4 by multiple 3'UTR-binding microRNAs that tune intracellular Mg2+ in hepatocytes.\",\n      \"evidence\": \"High-throughput miRNA 3'UTR binding assay with miRNA overexpression and intracellular Mg2+ measurement\",\n      \"pmids\": [\"40862638\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological contexts driving these miRNAs unknown\", \"In vivo relevance not tested\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Linked CNNM4 to cholangiocarcinoma aggressiveness and identified a NUPR1-associated ferroptosis axis as the mechanistic pathway.\",\n      \"evidence\": \"siRNA/shRNA and GalNAc-siRNA knockdown in vitro and in vivo, CAM invasion model, proteomics, and deferiprone/ZnPP rescue\",\n      \"pmids\": [\"40764063\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direction of CNNM4-NUPR1 regulation not fully mechanistically resolved\", \"Link between Mg2+ transport and iron/ferroptosis unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural mechanism by which CNNM4 transports Mg2+, how its CBS-pair/ATP-binding module gates transport, and how Mg2+ efflux is mechanistically coupled to downstream Ca2+, EGFR, mTOR, and ferroptosis pathways remain unresolved.\",\n      \"evidence\": \"No reconstituted transport or full-length structure in the timeline; CBS-domain crystallization and ATP-docking remain preliminary\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No high-resolution structure of full-length transporter\", \"ATP regulation of CBS domain unvalidated experimentally\", \"Transport stoichiometry and electrogenicity unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005215\", \"supporting_discovery_ids\": [1, 2, 9, 10]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-382551\", \"supporting_discovery_ids\": [1, 9, 10]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [4, 10]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [0, 11, 13]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [13]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"PRL phosphatases\", \"AP-1A (\\u03bc1A)\", \"AP-1B (\\u03bc1B)\", \"COX11\", \"IQCB1\", \"TRPV1\", \"TRPM7\", \"PCMT1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}