{"gene":"MANF","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":2003,"finding":"MANF (initially derived from a rat mesencephalic type-1 astrocyte cell line) is a secreted ~20 kDa protein that selectively protects nigral dopaminergic neurons in vitro, with greater selectivity than GDNF or BDNF at low-to-middle concentrations; contains two conserved domains and eight cysteines conserved from C. elegans to human.","method":"Recombinant protein production, in vitro neuronal survival assays comparing MANF, GDNF, and BDNF dose-response on dopaminergic vs. GABAergic/serotonergic neurons","journal":"Journal of molecular neuroscience : MN","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro functional assay with concentration-response, recombinant protein, but single lab and limited mechanistic depth","pmids":["12794311"],"is_preprint":false},{"year":2007,"finding":"Mouse ARMET/MANF is an 18 kDa soluble ER protein that, after signal-sequence cleavage, contains four intramolecular disulfide bonds including two in CXXC sequences; its UPR-dependent transcriptional induction is regulated by an ERSE-II element (identical to the HERP gene ERSE-II) in the MANF promoter.","method":"Western blotting, in vitro disulfide-bond characterization, reporter gene assay with MANF promoter deletion constructs","journal":"Cell structure and function","confidence":"High","confidence_rationale":"Tier 1 / Moderate — biochemical characterization of protein maturation (disulfide bonds) combined with reporter gene assay for promoter element; multiple orthogonal methods in one study","pmids":["17507765"],"is_preprint":false},{"year":2008,"finding":"ARMET/MANF protein is upregulated by multiple forms of ER stress in several cell lines and by cerebral ischemia in rat; it localizes to the ER and Golgi and is also secreted; siRNA-mediated silencing increases susceptibility to ER stress-induced death and paradoxically increases proliferation, while overexpression inhibits proliferation and improves viability under stress, establishing MANF as a secreted mediator of the adaptive UPR pathway.","method":"siRNA knockdown, overexpression, immunofluorescence (ER/Golgi localization), cell viability assays, immunohistochemistry in rat ischemia model","journal":"Experimental cell research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal gain- and loss-of-function with defined phenotypic readouts, direct localization, multiple cell lines and an in vivo model","pmids":["18561914"],"is_preprint":false},{"year":2009,"finding":"Crystal structures of human MANF and CDNF reveal that neither resembles known growth factors; the N-terminal domain is a saposin-like lipid-binding domain (suggesting membrane/lipid interaction), while the natively unfolded C-terminal domain of MANF contains a CKGC disulfide bridge analogous to reductases/disulfide isomerases, consistent with a role in ER stress response. Three residues (I10, E79, K88 in MANF) may account for functional differences between MANF and CDNF.","method":"X-ray crystallography (crystal structures of both proteins solved), structural bioinformatics","journal":"Protein engineering, design & selection : PEDS","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structures solved and validated; structural basis for bifunctionality established; independently consistent with NMR study (PMID 21047780)","pmids":["19258449"],"is_preprint":false},{"year":2010,"finding":"Solution NMR structure of human MANF shows the C-terminal domain (C-MANF) is homologous to the SAP domain of Ku70, a known inhibitor of pro-apoptotic Bax; cellular experiments confirm that both full-length MANF and C-MANF protect neurons intracellularly as efficiently as Ku70.","method":"NMR structure determination, cellular neuroprotection assays with MANF and C-MANF constructs","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — NMR structure with functional validation in cells; structural homology to Ku70 SAP domain experimentally supported by cellular assays","pmids":["21047780"],"is_preprint":false},{"year":2010,"finding":"Solution structure of mouse ARMET/MANF (by NMR relaxation and trypsin digestion) reveals an entirely α-helical two-domain architecture in which the N-terminal and C-terminal domains tumble independently; positive charges are dispersed across both domains and the linker, suggesting a negatively-charged ligand-binding mode.","method":"NMR solution structure, 15N relaxation experiments, trypsin digestion","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 1 / Moderate — NMR structure with dynamics characterization; independent of but consistent with crystal structure (PMID 19258449)","pmids":["20214902"],"is_preprint":false},{"year":2012,"finding":"The C-terminal RTDL sequence of MANF is required for both ER retention and stress-responsive secretion; KDEL receptor (KDELR) overexpression reduces MANF secretion only when RTDL is present; MANF binds to KDELRs at the plasma membrane surface (inhibited by a KDELR-interacting peptide), and surface KDELR levels increase after ER stress.","method":"Lentiviral vector constructs with/without RTDL, KDELR overexpression, peptide competition, immunofluorescence, neuroblastoma cells and primary cortical neurons","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal approaches (deletion mutants, overexpression, peptide competition, FACS/imaging); two cell systems","pmids":["23255601"],"is_preprint":false},{"year":2013,"finding":"In Drosophila, only full-length MANF containing both N-terminal saposin-like and C-terminal SAP domains rescues larval lethality of DmManf null mutants; neither domain alone (even co-expressed) is sufficient; deleting the signal peptide or mutating the CXXC motif in the C-terminal domain destroys activity; positively charged surface residues and the C-terminal ER retention signal are necessary for rescue when expression is restricted.","method":"Transgenic rescue experiments in DmManf mutant background, domain-deletion and point-mutation constructs, in vitro sympathetic neuron survival assay","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vivo genetic rescue with multiple structure–function mutants; functional conservation confirmed in mammalian neuronal assay","pmids":["24019940"],"is_preprint":false},{"year":2014,"finding":"MANF is indispensable for pancreatic β-cell proliferation and survival in mice: MANF-deficient mice develop progressive postnatal loss of β-cell mass causing diabetes, with chronic UPR activation in islets; recombinant MANF enhances β-cell proliferation in vitro; MANF overexpression in diabetic mouse pancreas enhances β-cell regeneration.","method":"MANF knockout mice (loss-of-function), in vitro proliferation assays with recombinant MANF, in vivo pancreatic MANF overexpression in diabetic mice, UPR marker analysis","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal loss- and gain-of-function in vivo and in vitro with specific phenotypic readouts; replicated across models","pmids":["24726366"],"is_preprint":false},{"year":2014,"finding":"Reduced chaperone (Hsc70) activity with aging leads to decreased association of mutant TBP with XBP1s, reducing XBP1s-driven transcription of MANF; overexpression of MANF ameliorates Purkinje cell degeneration via PKC-dependent signaling; Hsc70 overexpression restores TBP-XBP1s interaction and MANF transcription.","method":"SCA17 knockin mice with tamoxifen-inducible expression, co-IP (TBP-XBP1s interaction), Hsc70 overexpression, PKC inhibitor studies, viral MANF overexpression","journal":"Neuron","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic epistasis in vivo, Co-IP for interaction, pharmacological pathway dissection, multiple orthogonal methods","pmids":["24462098"],"is_preprint":false},{"year":2016,"finding":"PDGF-like signaling induces MANF expression in innate immune cells of damaged retina (in both flies and mice); MANF promotes alternative (M2) activation of innate immune cells, enhancing neuroprotection and tissue repair, and improves photoreceptor replacement therapy success.","method":"Genetic epistasis in Drosophila retina damage model, mouse retinal damage model, MANF gain/loss-of-function, immune cell phenotyping, photoreceptor transplantation assay","journal":"Science (New York, N.Y.)","confidence":"High","confidence_rationale":"Tier 2 / Strong — conserved pathway established in two organisms (fly and mouse) with genetic and pharmacological tools; multiple phenotypic readouts","pmids":["27365452"],"is_preprint":false},{"year":2016,"finding":"In Drosophila, Manf shows genetic interactions with GRP78 (Hsc70-3), PERK (PEK), and XBP1 homologs, placing DmManf as a regulator of the UPR; DmManf expression is upregulated by pharmacological ER stress inducers, a response conserved between Drosophila and mammals.","method":"Genetic interaction assays (double mutant/RNAi combinations), drug-induced ER stress, reporter gene expression","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis in Drosophila; conservation supported by parallel mammalian data; single lab","pmids":["26975047"],"is_preprint":false},{"year":2017,"finding":"MANF is highly enriched in hypothalamic nuclei and its expression is upregulated by fasting; increasing hypothalamic MANF promotes hyperphagia, decreasing it causes hypophagia; mechanistically, MANF enhances ER localization and activity of PIP4k2b kinase, thereby triggering hypothalamic insulin resistance.","method":"Viral vector-mediated overexpression and knockdown in mouse hypothalamus, food intake/body weight measurements, PIP4k2b co-localization and activity assays","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo gain- and loss-of-function with specific signaling pathway dissection; single lab but multiple approaches","pmids":["28924165"],"is_preprint":false},{"year":2017,"finding":"MANF-deficient neural stem cells show no proliferation defect but exhibit impaired neurite extension upon differentiation; in vivo MANF deletion causes slower neuronal migration and impaired neurite outgrowth, preceded by reduced de novo protein synthesis and constitutively activated UPR pathways.","method":"MANF knockout NSC cultures, in utero electroporation/cortical migration assay, neurite outgrowth measurement, protein synthesis assay, UPR pathway marker analysis","journal":"eNeuro","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo and in vitro loss-of-function with mechanistic UPR pathway readouts; multiple orthogonal methods","pmids":["29082311"],"is_preprint":false},{"year":2017,"finding":"MANF promotes differentiation and migration of neural progenitor cells (NPCs) in vitro through activation of STAT3 and ERK1/2; in SVZ explants, MANF overexpression facilitates cell migration via STAT3 and ERK1/2; in vivo, intracerebroventricular MANF promotes migration of DCX+ neuroblasts after stroke.","method":"NSC/NPC culture with recombinant MANF, SVZ explant migration assay, STAT3/ERK phosphorylation Western blot, rat cortical stroke model with ICV injection","journal":"Molecular therapy : the journal of the American Society of Gene Therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro signaling assays plus in vivo model; pathway identified by Western blot; single lab","pmids":["29050872"],"is_preprint":false},{"year":2018,"finding":"In human β-cells, exogenous recombinant MANF reduces cytokine-induced cell death by ~38%; the protective effect is associated with repression of NF-κB signaling and amelioration of ER stress; MANF knockdown aggravates ER stress after cytokine challenge.","method":"Primary human islets + recombinant MANF, siRNA knockdown in EndoC-βH1 cells, cell viability, global transcriptomics, NF-κB pathway analysis","journal":"Diabetologia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal gain/loss-of-function in human cells with defined pathway readout; single lab","pmids":["30032427"],"is_preprint":false},{"year":2019,"finding":"The SAP domain of MANF selectively associates with the nucleotide-binding domain (NBD) of ADP-bound BiP; crystal structures show the SAP domain engages the cleft between NBD subdomains Ia and IIa, stabilizing ADP-bound conformation and clashing with the ATP-bound interdomain linker; MANF inhibits both ADP release from and ATP binding to BiP, thereby inhibiting client release; cells lacking MANF have fewer ER stress-induced BiP-containing high-molecular-weight complexes.","method":"Crystal structures of MANF SAP domain–BiP NBD complex, in vitro nucleotide exchange assays, MANF knockout cells, size-exclusion chromatography of BiP complexes","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus in vitro enzymatic assay plus cell-based validation; multiple orthogonal methods; single lab but rigorous","pmids":["30710085"],"is_preprint":false},{"year":2019,"finding":"MANF deficiency in flies causes enhanced inflammation and shorter lifespan; MANF deficiency in mice causes inflammatory phenotypes and progressive liver damage (fibrosis, steatosis); immune cell-derived MANF specifically protects against liver inflammation and fibrosis, while hepatocyte-derived MANF prevents hepatosteatosis; liver rejuvenation by heterochronic parabiosis depends on MANF.","method":"Drosophila MANF overexpression/KO, MANF heterozygous mice, tissue-specific MANF deletion (immune cells vs. hepatocytes), heterochronic parabiosis, recombinant MANF supplementation","journal":"Nature metabolism","confidence":"High","confidence_rationale":"Tier 2 / Strong — cell-type-specific KOs, parabiosis, recombinant protein rescue, conserved across two organisms; multiple orthogonal methods","pmids":["31489403"],"is_preprint":false},{"year":2020,"finding":"Brain-specific MANF deletion exacerbates ethanol-induced neuronal apoptosis and ER stress; blocking ER stress abrogates the detrimental effects of MANF deficiency on ethanol-induced apoptosis, placing MANF upstream of ER stress in ethanol neurotoxicity; in a tunicamycin-induced neurodegeneration model, MANF deficiency potentiates ER stress and neurodegeneration.","method":"CNS-specific Manf knockout mice, tunicamycin and ethanol in vivo models, ER stress marker analysis, RNA-seq","journal":"Neurobiology of disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with pharmacological epistasis (ER stress inhibitor rescue); pathway positioning established; single lab","pmids":["33296727"],"is_preprint":false},{"year":2020,"finding":"MANF loss causes prolonged activation of the IRE1α branch of the UPR (and in aged mice additionally PERK and ATF6 branches) without causing dopaminergic neuron loss in the substantia nigra or behavioral changes; cortical neurons lacking MANF are more vulnerable to additional ER stress in vitro, indicating endogenous MANF is required for neuronal ER homeostasis maintenance.","method":"Brain-specific MANF knockout mice, UPR marker analysis at multiple ages, stereological neuron counting in SN, dopamine measurement, cortical neuron cultures with ER stress challenge","journal":"eNeuro","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with comprehensive pathway and phenotypic analysis in vivo and in vitro; multiple UPR branches characterized","pmids":["32005751"],"is_preprint":false},{"year":2020,"finding":"Neuroplastin (NPTN) is a cell-surface receptor for MANF; biochemical analysis shows physiological interaction between MANF and NPTN on the cell surface; MANF binding to NPTN suppresses NF-κB signaling, mitigating inflammatory response and apoptosis.","method":"Co-immunoprecipitation, surface binding assays, NPTN knockdown/overexpression, NF-κB reporter assays","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Co-IP and surface binding with functional NF-κB readout; single lab; mechanistic pathway proposed with supporting experiments","pmids":["33299977"],"is_preprint":false},{"year":2020,"finding":"MANF regulates neurite outgrowth by activating Akt/mTOR and Erk/mTOR signaling pathways; MANF KO N2a cells fail to grow neurites on RA stimulation, and this is rescued by MANF overexpression or exogenous MANF; pharmacological blockade of Akt, Erk, or mTOR eliminates the MANF-promoting effect on neurite outgrowth.","method":"CRISPR/Cas9 MANF KO in N2a cells, siRNA in SH-SY5Y, adenoviral MANF overexpression, pharmacological inhibitors of Akt/Erk/mTOR, Western blot for pathway activation","journal":"Frontiers in molecular neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR KO, siRNA, overexpression, and pharmacological epistasis; multiple cell lines; single lab","pmids":["33071755"],"is_preprint":false},{"year":2021,"finding":"MANF promotes neuronal survival as a general UPR regulator affecting multiple UPR pathways simultaneously, independently of its role as a GRP78 cofactor; MANF interacts with a conserved set of 15 ER proteins including GRP78, GRP170, PDIA1, and PDIA6; ATP binding to MANF (confirmed by MST and NMR) blocks the MANF-GRP78 interaction; antiapoptotic activity of MANF mutants in neurons reveals divergent roles as GRP78 cofactor vs. antiapoptotic UPR regulator.","method":"AP-MS interactome from two mammalian cell lines, microscale thermophoresis (ATP binding), NMR, neuronal apoptosis assays with MANF mutants","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — biochemical ATP-binding assay (MST + NMR) plus interactome plus neuronal functional assays; multiple orthogonal methods; single lab","pmids":["33460650"],"is_preprint":false},{"year":2021,"finding":"Loss-of-function mutations in the MANF gene in humans cause childhood-onset diabetes with neurodevelopmental disorder; MANF knockout in human embryonic stem cell-derived β-cells induces mild ER stress and impairs insulin-processing capacity; upon implantation in diabetic mice, MANF-KO grafts show elevated ER stress and functional failure.","method":"Human genetic (homozygous LOF mutations), hESC MANF knockout, β-cell differentiation, insulin processing assay, xenotransplantation in immunocompromised diabetic mice","journal":"Diabetes","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — human genetics validated mechanistically with hESC KO and in vivo xenograft; multiple orthogonal approaches","pmids":["33500254"],"is_preprint":false},{"year":2021,"finding":"XBP1s (spliced XBP1) binds to ERSE-I in the MANF promoter to promote MANF transcription; MANF in turn interacts with XBP1s to enhance its own expression in a positive feedback loop; IRE1α endonuclease inhibition or XBP1s knockdown attenuates MANF expression.","method":"ChIP, promoter reporter assay, XBP1s overexpression and siRNA knockdown, IRE1α inhibitor, co-IP of MANF with XBP1s","journal":"The international journal of biochemistry & cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and reporter gene assay plus Co-IP; multiple methods; single lab","pmids":["29649564"],"is_preprint":false},{"year":2021,"finding":"Liver-specific MANF overexpression protects against high-fat diet-induced obesity and promotes browning (beige adipogenesis) of inguinal WAT; mechanistically, MANF directly promotes white adipocyte browning via the p38 MAPK pathway; p38 MAPK blockade abolishes MANF-induced browning; liver-specific MANF KO impairs WAT browning and worsens diet-induced obesity.","method":"Liver-specific overexpression and KO mouse models, p38 MAPK inhibitor, primary white adipocyte culture with recombinant MANF, thermogenesis markers","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal liver-specific OE and KO in vivo, pharmacological epistasis, in vitro mechanistic confirmation; multiple orthogonal approaches","pmids":["33856409"],"is_preprint":false},{"year":2022,"finding":"MANF is transcriptionally induced in cardiomyocytes by liganded estrogen receptor β and inhibited by androgen receptor; ICI (immune checkpoint inhibitor) treatment reduces serum estradiol, decreasing cardiac MANF/HSPA5; heart-specific MANF deletion amplifies ICI myocarditis, and recombinant MANF protein attenuates it, establishing a causal role.","method":"Genetic cardiac-specific Manf deletion, recombinant MANF administration, ERβ agonist and androgen depletion in vivo, transcriptional regulation analysis","journal":"Science translational medicine","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO, recombinant protein rescue, and pharmacological pathway dissection; multiple orthogonal methods in one study","pmids":["36322628"],"is_preprint":false},{"year":2023,"finding":"MANF directly interacts with the ER transmembrane UPR sensor IRE1α; wild-type MANF (but not its IRE1α-binding-deficient mutant) decreases IRE1α oligomerization and phosphorylation, reduces XBP1 splicing, ATF6 and TXNIP levels, and protects neurons from ER stress-induced death; MANF-IRE1α interaction (not MANF-BiP interaction) is required for protection of dopamine neurons in a Parkinson's disease animal model.","method":"Co-IP of MANF with IRE1α, binding-interface mutagenesis, IRE1α oligomerization/phosphorylation assays, XBP1 splicing reporter, neuronal apoptosis assays, 6-OHDA rat PD model with MANF mutant constructs","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — direct protein interaction mapped to binding interface, multiple UPR functional readouts, mutant dissection of MANF-IRE1α vs. MANF-BiP, in vivo PD model validation","pmids":["36739529"],"is_preprint":false},{"year":2023,"finding":"MANF overexpression in kidney tubular cells stimulates autophagy/mitophagy and mitochondrial biogenesis through p-AMPK enhancement, clears mutant UMOD aggregates, and suppresses cGAS-STING activation; genetic ablation of tubular MANF worsens autophagy suppression and kidney fibrosis in a ADTKD-UMOD mouse model.","method":"Inducible tubular MANF overexpression and knockout in ADTKD-UMOD mice, autophagy/mitophagy markers, AMPK phosphorylation, cGAS-STING pathway analysis, mutant UMOD clearance","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal in vivo gain- and loss-of-function with mechanistic pathway identification (AMPK, autophagy, cGAS-STING); disease-relevant model","pmids":["37838725"],"is_preprint":false},{"year":2023,"finding":"MANF brakes macrophage TLR4 signaling by competitively binding S100A8, preventing S100A8/A9 heterodimer formation and thereby inhibiting S100A8/A9-mediated TLR4-NF-κB activation; myeloid-specific MANF KO increases hepatic Ly6Chigh pro-inflammatory macrophages and worsens hepatic fibrosis.","method":"Myeloid-specific MANF KO mice, competitive binding assay (MANF vs. S100A9 for S100A8), Co-IP, macrophage phenotyping, TLR4-NF-κB pathway analysis, recombinant MANF treatment","journal":"Acta pharmaceutica Sinica. B","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO plus direct competitive binding assay plus signaling pathway validation; multiple orthogonal methods","pmids":["37799387"],"is_preprint":false},{"year":2024,"finding":"Under glucose starvation, SENP1-mediated de-SUMOylation of MANF inhibits its nuclear translocation and increases cytoplasmic/mitochondrial MANF; mitochondrial MANF binds to PRKN (Parkin) E3 ligase, alleviates oxidative inhibition of PRKN's RING II domain via its CXXC motif, restores PRKN E3 ligase activity, and thereby mediates mitophagy to promote breast cancer cell survival.","method":"SENP1 manipulation (de-SUMOylation), Co-IP of MANF-PRKN, PRKN ubiquitin ligase activity assay, CXXC motif mutagenesis, mitophagy assays, MANF subcellular fractionation","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, enzymatic activity restoration, mutagenesis of CXXC motif; multiple methods; single lab","pmids":["39147386"],"is_preprint":false},{"year":2013,"finding":"ARMET/MANF interacts with mutant matrilin-3 (but not COMP) in chondrocyte ER stress models of skeletal dysplasia, establishing substrate specificity; both ARMET and CRELD2 are secreted into ECM following ER stress; substrate-trapping experiments confirmed CRELD2 but not ARMET has PDI-like activity.","method":"Cell and mouse models of chondrodysplasia, co-IP/substrate trapping, immunohistochemistry, Western blotting; genotype-specific expression analysis","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — substrate specificity shown by Co-IP and substrate trapping; genotype-specific interaction validated; single lab","pmids":["23956175"],"is_preprint":false},{"year":2017,"finding":"THBS1 (thrombospondin 1) maintains MANF expression in β-cells exposed to cytokines or thapsigargin, and this THBS1-MANF axis prevents BIM-dependent mitochondrial apoptosis; prolonged stress leads to THBS1 and MANF degradation and loss of this prosurvival mechanism.","method":"THBS1 overexpression/silencing in rat/mouse/human β-cells, Western blot for MANF and BIM, apoptosis assays, ER stress induction","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pathway epistasis (THBS1→MANF→BIM) with gain/loss-of-function and specific apoptotic readout; multiple species tested; single lab","pmids":["28698383"],"is_preprint":false},{"year":2022,"finding":"CDNF and MANF display functional redundancy in skeletal muscle (CDNF deficiency increases UPR activation, aggravated by combined MANF loss) but not in brain; neither single nor combined brain-specific KO causes dopaminergic neuron degeneration, showing tissue-specific UPR regulation.","method":"Cdnf−/−, Manf−/−, and double KO mice, UPR markers in multiple tissues, dopaminergic neuron counting","journal":"Cellular and molecular life sciences : CMLS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — double KO with tissue-specific phenotypic analysis; defines functional redundancy/divergence; single lab","pmids":["35129674"],"is_preprint":false}],"current_model":"MANF is a bifunctional ER-resident/secreted protein whose N-terminal saposin-like domain mediates lipid/membrane interaction and extracellular neurotrophic activity, while its natively unfolded C-terminal SAP/CXXC domain acts intracellularly as a nucleotide-exchange inhibitor of the Hsp70 chaperone BiP (stabilizing ADP-bound BiP–client complexes) and as a direct binding partner and negative regulator of the UPR sensor IRE1α (reducing its oligomerization, phosphorylation, and XBP1 splicing); extracellularly, MANF binds the cell-surface receptor Neuroplastin to suppress NF-κB-mediated inflammation, competes with S100A9 for S100A8 to brake TLR4–NF-κB macrophage activation, and is retained in the ER via its C-terminal RTDL sequence through interaction with KDEL receptors whose surface levels rise after ER stress; loss of MANF in mice causes chronic UPR activation, progressive β-cell death and diabetes, impaired neurite outgrowth, and tissue-specific pathologies (liver fibrosis, outer hair cell death, hearing loss), while its protective signaling engages PI3K/Akt/mTOR, ERK/mTOR, Nrf2, and AMPK pathways depending on context."},"narrative":{"mechanistic_narrative":"MANF is a bifunctional ER-resident and secreted protein that maintains endoplasmic reticulum proteostasis and confers cytoprotection across neuronal, pancreatic, hepatic, immune, and cardiac tissues, functioning as a secreted mediator of the adaptive unfolded protein response (UPR) [PMID:18561914, PMID:24726366]. Its two-domain architecture underlies this bifunctionality: an N-terminal saposin-like lipid-binding domain and a natively unfolded C-terminal SAP/CXXC domain homologous to the Bax inhibitor Ku70, with both domains required for full activity in vivo [PMID:19258449, PMID:21047780, PMID:24019940]. Intracellularly, the SAP domain binds the nucleotide-binding domain of ADP-bound BiP, stabilizing the ADP conformation and inhibiting client release, and ATP binding to MANF disrupts this interaction [PMID:30710085, PMID:33460650]; MANF also directly binds the UPR sensor IRE1α, reducing its oligomerization and phosphorylation and dampening XBP1 splicing, and this IRE1α interaction — distinct from BiP binding — is required for dopaminergic neuroprotection [PMID:36739529]. MANF expression is induced by ER stress through ERSE elements and an XBP1s-driven positive feedback loop [PMID:17507765, PMID:29649564]. The C-terminal RTDL signal mediates ER retention and stress-responsive secretion via KDEL receptors [PMID:23255601]. Extracellularly, MANF acts through the cell-surface receptor Neuroplastin to suppress NF-κB signaling and competes with S100A9 for S100A8 to brake TLR4–NF-κB macrophage activation, promoting anti-inflammatory immune phenotypes and tissue repair [PMID:27365452, PMID:33299977, PMID:37799387], and engages PI3K/Akt/mTOR, ERK/mTOR, STAT3, AMPK, and p38 MAPK signaling in a context-dependent manner to promote neurite outgrowth, autophagy, and metabolic regulation [PMID:29050872, PMID:33071755, PMID:37838725, PMID:33856409]. Loss of MANF causes chronic UPR activation with progressive β-cell death and diabetes, impaired neurite outgrowth and neuronal migration, and tissue-specific liver and inflammatory pathology [PMID:24726366, PMID:29082311, PMID:31489403]. Homozygous loss-of-function MANF mutations cause childhood-onset diabetes with a neurodevelopmental disorder [PMID:33500254].","teleology":[{"year":2003,"claim":"Established MANF as a secreted factor with selective trophic activity, defining the founding functional question of how a non-canonical protein protects specific neuron populations.","evidence":"Recombinant protein with in vitro dopaminergic neuron survival assays comparing MANF, GDNF, and BDNF","pmids":["12794311"],"confidence":"Medium","gaps":["No receptor or molecular mechanism for selectivity identified","Single cell-line-derived source","No structural basis for activity"]},{"year":2008,"claim":"Positioned MANF within the UPR by showing ER stress induces it and its loss/gain reciprocally modulates stress survival, recasting the trophic factor as a UPR effector.","evidence":"siRNA knockdown, overexpression, ER/Golgi immunofluorescence, viability assays, and rat ischemia model","pmids":["18561914"],"confidence":"High","gaps":["Molecular target within the UPR unknown","Mechanism of cytoprotection not defined","Secreted vs. intracellular roles not separated"]},{"year":2010,"claim":"Resolved the structural basis of MANF's bifunctionality, distinguishing a lipid-binding N-domain from a Ku70-like SAP/CXXC C-domain implicated in apoptosis suppression and redox activity.","evidence":"X-ray crystallography and NMR solution structures with cellular neuroprotection assays of full-length and C-terminal constructs","pmids":["19258449","21047780","20214902"],"confidence":"High","gaps":["No binding partner for either domain identified","Mechanism of SAP-domain neuroprotection unresolved","Functional ligand of the saposin-like domain unknown"]},{"year":2012,"claim":"Defined how MANF balances ER retention against secretion, showing the RTDL signal and KDEL receptors gate stress-responsive release.","evidence":"RTDL deletion constructs, KDELR overexpression, peptide competition, and imaging in neuroblastoma and primary cortical neurons","pmids":["23255601"],"confidence":"High","gaps":["Surface KDELR-MANF signaling consequences not defined","Trigger linking ER stress to surface KDELR increase unclear"]},{"year":2013,"claim":"Demonstrated through in vivo genetic rescue that both MANF domains are jointly required, establishing the protein acts as an integrated bifunctional unit rather than two separable activities.","evidence":"Transgenic rescue of DmManf null with domain-deletion and CXXC point mutants plus mammalian neuron assays","pmids":["24019940"],"confidence":"High","gaps":["Molecular partners of each domain not identified in this system","Mechanistic basis of domain interdependence unresolved"]},{"year":2014,"claim":"Identified MANF as essential for pancreatic β-cell proliferation and survival, linking its UPR-modulating function to a defined organ phenotype (diabetes).","evidence":"MANF knockout mice, recombinant MANF proliferation assays, and pancreatic overexpression in diabetic mice","pmids":["24726366"],"confidence":"High","gaps":["Molecular mechanism of proliferative effect not defined","Secreted vs. intracellular contributions not separated"]},{"year":2017,"claim":"Extended MANF function beyond cytoprotection to neural progenitor migration/differentiation and central metabolic control, revealing context-dependent signaling outputs.","evidence":"MANF KO NSCs, in utero electroporation, STAT3/ERK Western blots, and hypothalamic viral gain/loss-of-function with PIP4k2b assays","pmids":["29082311","29050872","28924165"],"confidence":"Medium","gaps":["Receptor mediating progenitor signaling not defined","Connection between ER role and surface signaling unclear","Generality across cell types untested"]},{"year":2019,"claim":"Solved the central intracellular mechanism by showing the SAP domain binds ADP-bound BiP's nucleotide-binding domain and inhibits nucleotide exchange, stabilizing chaperone-client complexes.","evidence":"Crystal structures of MANF SAP–BiP NBD complex, in vitro nucleotide exchange assays, and BiP complex analysis in MANF-null cells","pmids":["30710085"],"confidence":"High","gaps":["Cellular consequence of stabilized BiP complexes incompletely defined","Whether BiP regulation accounts for all protective effects unresolved"]},{"year":2020,"claim":"Identified Neuroplastin as a cell-surface MANF receptor coupling extracellular MANF to NF-κB suppression, providing a receptor for the secreted anti-inflammatory function.","evidence":"Co-immunoprecipitation, surface binding assays, NPTN manipulation, and NF-κB reporter assays","pmids":["33299977"],"confidence":"Medium","gaps":["Single lab, no reciprocal in vivo receptor validation","Downstream signaling between NPTN and NF-κB not mapped"]},{"year":2021,"claim":"Established that MANF is a broad UPR regulator with an ER interactome beyond BiP, that ATP binding gates the BiP interaction, and that human LOF mutations cause a Mendelian disease, validating its physiological essentiality.","evidence":"AP-MS interactome, microscale thermophoresis/NMR ATP binding, neuronal mutant assays; human genetics with hESC β-cell KO and xenotransplantation","pmids":["33460650","33500254"],"confidence":"High","gaps":["Functional roles of most interactome partners undefined","How distinct activities map to disease phenotypes unresolved"]},{"year":2023,"claim":"Defined direct MANF–IRE1α binding as a BiP-independent mechanism that restrains the IRE1α UPR branch and is specifically required for dopaminergic neuroprotection in vivo.","evidence":"Co-IP, binding-interface mutagenesis, IRE1α oligomerization/phosphorylation and XBP1 splicing assays, and 6-OHDA PD model with MANF mutants","pmids":["36739529"],"confidence":"High","gaps":["Relative in vivo weighting of IRE1α vs. BiP mechanisms across tissues unresolved","Structural basis of MANF-IRE1α interface not solved"]},{"year":2024,"claim":"Revealed a non-canonical mitochondrial role in which de-SUMOylated MANF restores Parkin E3 ligase activity via its CXXC motif to drive mitophagy, expanding MANF function beyond the ER.","evidence":"SENP1 manipulation, MANF-PRKN Co-IP, ubiquitin ligase activity and mitophagy assays, CXXC mutagenesis in breast cancer cells","pmids":["39147386"],"confidence":"Medium","gaps":["Single lab; physiological generality of mitochondrial MANF untested","Trafficking route to mitochondria unclear"]},{"year":null,"claim":"How MANF's distinct intracellular (BiP, IRE1α, mitochondrial Parkin) and extracellular (Neuroplastin, S100A8) activities are coordinated and weighted within a given cell type to produce tissue-specific protection remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model integrating ER, secreted, and mitochondrial functions","Receptor-to-signaling cascades incompletely mapped","Determinants of tissue-specific phenotype selection unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[16,27]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[16,22]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[3]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[22,16]},{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[20]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[1,2,16]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[2]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[2,6]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6,20]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[30]}],"pathway":[{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[2,16,27]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[16,22]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[10,20,29]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[28,30]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[21,14,25]}],"complexes":[],"partners":["HSPA5","ERN1","NPTN","S100A8","XBP1","PRKN","MATN3"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P55145","full_name":"Mesencephalic astrocyte-derived neurotrophic factor","aliases":["Arginine-rich protein","Protein ARMET"],"length_aa":182,"mass_kda":20.7,"function":"Selectively promotes the survival of dopaminergic neurons of the ventral mid-brain (PubMed:12794311). Modulates GABAergic transmission to the dopaminergic neurons of the substantia nigra (By similarity). Enhances spontaneous, as well as evoked, GABAergic inhibitory postsynaptic currents in dopaminergic neurons (By similarity). Inhibits cell proliferation and endoplasmic reticulum (ER) stress-induced cell death (PubMed:18561914, PubMed:22637475, PubMed:29497057, PubMed:36739529). Retained in the ER/sarcoplasmic reticulum (SR) through association with the endoplasmic reticulum chaperone protein HSPA5 under normal conditions (PubMed:22637475). Stabilizes HSPA5/BiP in its substrate-bound ADP state, which facilitates HSPA5/BiP incorporation into chaperone-client complexes during endoplasmic reticulum stress, its interaction with HSPA5/BiP inhibits ATP binding to HSPA5/BiP and subsequent nucleotide exchange (By similarity). As a result acts as a repressor of the unfolded protein response (UPR) pathway (By similarity). Up-regulated and secreted by the ER/SR in response to ER stress and hypoxia (PubMed:22637475). Following secretion by the ER/SR, directly binds to 3-O-sulfogalactosylceramide, a lipid sulfatide in the outer cell membrane of target cells (PubMed:29497057). Sulfatide binding promotes its cellular uptake by endocytosis, and is required for its role in alleviating ER stress and cell toxicity under hypoxic and ER stress conditions (PubMed:29497057). Essential for embryonic lung development (By similarity). Required for the correct postnatal temporal and structural development of splenic white pulp (By similarity). Required for the repair-associated myeloid response in skeletal muscle, acts as a regulator of phenotypic transition towards prorepair macrophages in response to muscle injury and as a result limits excessive proinflammatory signaling (By similarity). Represses RELA expression and therefore NF-kB signaling in the myocardium, as a result limits macrophage infiltration of injured tissue and M1 macrophage differentiation in response to myocardial injury (By similarity). Required for endochondral ossification in long bones and the skull during postnatal development (By similarity)","subcellular_location":"Secreted; Endoplasmic reticulum lumen; Sarcoplasmic reticulum lumen","url":"https://www.uniprot.org/uniprotkb/P55145/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MANF","classification":"Not Classified","n_dependent_lines":485,"n_total_lines":1208,"dependency_fraction":0.4014900662251656},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"STK25","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/MANF","total_profiled":1310},"omim":[{"mim_id":"620651","title":"DIABETES, DEAFNESS, DEVELOPMENTAL DELAY, AND SHORT STATURE SYNDROME; DDDS","url":"https://www.omim.org/entry/620651"},{"mim_id":"611233","title":"ARGININE-RICH PROTEIN MUTATED IN EARLY STAGE TUMORS-LIKE 1; ARMETL1","url":"https://www.omim.org/entry/611233"},{"mim_id":"601916","title":"MESENCEPHALIC ASTROCYTE-DERIVED NEUROTROPHIC FACTOR; MANF","url":"https://www.omim.org/entry/601916"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Endoplasmic reticulum","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MANF"},"hgnc":{"alias_symbol":["ARP"],"prev_symbol":["ARMET"]},"alphafold":{"accession":"P55145","domains":[{"cath_id":"1.10.225.10","chopping":"30-117","consensus_level":"high","plddt":93.2456,"start":30,"end":117},{"cath_id":"1.10.720","chopping":"131-180","consensus_level":"high","plddt":80.8856,"start":131,"end":180}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P55145","model_url":"https://alphafold.ebi.ac.uk/files/AF-P55145-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P55145-F1-predicted_aligned_error_v6.png","plddt_mean":81.69},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MANF","jax_strain_url":"https://www.jax.org/strain/search?query=MANF"},"sequence":{"accession":"P55145","fasta_url":"https://rest.uniprot.org/uniprotkb/P55145.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P55145/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P55145"}},"corpus_meta":[{"pmid":"12794311","id":"PMC_12794311","title":"MANF: a new mesencephalic, astrocyte-derived neurotrophic factor with selectivity for dopaminergic neurons.","date":"2003","source":"Journal of molecular neuroscience : MN","url":"https://pubmed.ncbi.nlm.nih.gov/12794311","citation_count":295,"is_preprint":false},{"pmid":"18561914","id":"PMC_18561914","title":"Armet, a UPR-upregulated protein, inhibits cell proliferation and ER stress-induced cell death.","date":"2008","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/18561914","citation_count":237,"is_preprint":false},{"pmid":"27365452","id":"PMC_27365452","title":"Immune modulation by MANF promotes tissue repair and regenerative success in the retina.","date":"2016","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/27365452","citation_count":194,"is_preprint":false},{"pmid":"24726366","id":"PMC_24726366","title":"MANF is indispensable for the proliferation and survival of pancreatic β cells.","date":"2014","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/24726366","citation_count":170,"is_preprint":false},{"pmid":"18718866","id":"PMC_18718866","title":"MANF is widely expressed in mammalian tissues and differently regulated after ischemic and epileptic insults in rodent brain.","date":"2008","source":"Molecular and cellular neurosciences","url":"https://pubmed.ncbi.nlm.nih.gov/18718866","citation_count":158,"is_preprint":false},{"pmid":"17507765","id":"PMC_17507765","title":"ARMET is a soluble ER protein induced by the unfolded protein response via ERSE-II element.","date":"2007","source":"Cell structure and function","url":"https://pubmed.ncbi.nlm.nih.gov/17507765","citation_count":156,"is_preprint":false},{"pmid":"20186704","id":"PMC_20186704","title":"Novel CDNF/MANF family of neurotrophic factors.","date":"2010","source":"Developmental neurobiology","url":"https://pubmed.ncbi.nlm.nih.gov/20186704","citation_count":150,"is_preprint":false},{"pmid":"27425895","id":"PMC_27425895","title":"Unconventional neurotrophic factors CDNF and MANF: Structure, physiological functions and therapeutic potential.","date":"2016","source":"Neurobiology of disease","url":"https://pubmed.ncbi.nlm.nih.gov/27425895","citation_count":144,"is_preprint":false},{"pmid":"23255601","id":"PMC_23255601","title":"Mesencephalic astrocyte-derived neurotrophic factor (MANF) secretion and cell surface binding are modulated by KDEL receptors.","date":"2012","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23255601","citation_count":124,"is_preprint":false},{"pmid":"21047780","id":"PMC_21047780","title":"Mesencephalic astrocyte-derived neurotrophic factor (MANF) has a unique mechanism to rescue apoptotic neurons.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21047780","citation_count":121,"is_preprint":false},{"pmid":"31489403","id":"PMC_31489403","title":"MANF regulates metabolic and immune homeostasis in ageing and protects against liver damage.","date":"2019","source":"Nature metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/31489403","citation_count":110,"is_preprint":false},{"pmid":"19258449","id":"PMC_19258449","title":"The structure of the conserved neurotrophic factors MANF and CDNF explains why they are bifunctional.","date":"2009","source":"Protein engineering, design & selection : PEDS","url":"https://pubmed.ncbi.nlm.nih.gov/19258449","citation_count":105,"is_preprint":false},{"pmid":"19773801","id":"PMC_19773801","title":"Induction profile of MANF/ARMET by cerebral ischemia and its implication for neuron protection.","date":"2009","source":"Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/19773801","citation_count":100,"is_preprint":false},{"pmid":"24462098","id":"PMC_24462098","title":"Age-dependent decrease in chaperone activity impairs MANF expression, leading to Purkinje cell degeneration in inducible SCA17 mice.","date":"2014","source":"Neuron","url":"https://pubmed.ncbi.nlm.nih.gov/24462098","citation_count":92,"is_preprint":false},{"pmid":"30710085","id":"PMC_30710085","title":"MANF antagonizes nucleotide exchange by the endoplasmic reticulum chaperone BiP.","date":"2019","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/30710085","citation_count":90,"is_preprint":false},{"pmid":"22898306","id":"PMC_22898306","title":"MANF regulates dopaminergic neuron development in larval zebrafish.","date":"2012","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/22898306","citation_count":84,"is_preprint":false},{"pmid":"25678626","id":"PMC_25678626","title":"Armet is an effector protein mediating aphid-plant interactions.","date":"2015","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/25678626","citation_count":81,"is_preprint":false},{"pmid":"29050872","id":"PMC_29050872","title":"MANF Promotes Differentiation and Migration of Neural Progenitor Cells with Potential Neural Regenerative Effects in Stroke.","date":"2017","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/29050872","citation_count":77,"is_preprint":false},{"pmid":"20685313","id":"PMC_20685313","title":"Widespread cortical expression of MANF by AAV serotype 7: localization and protection against ischemic brain injury.","date":"2010","source":"Experimental neurology","url":"https://pubmed.ncbi.nlm.nih.gov/20685313","citation_count":74,"is_preprint":false},{"pmid":"30032427","id":"PMC_30032427","title":"MANF protects human pancreatic beta cells against stress-induced cell death.","date":"2018","source":"Diabetologia","url":"https://pubmed.ncbi.nlm.nih.gov/30032427","citation_count":73,"is_preprint":false},{"pmid":"23956175","id":"PMC_23956175","title":"Armet/Manf and Creld2 are components of a specialized ER stress response provoked by inappropriate formation of disulphide bonds: implications for genetic skeletal diseases.","date":"2013","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23956175","citation_count":71,"is_preprint":false},{"pmid":"30760285","id":"PMC_30760285","title":"Mesencephalic astrocyte-derived neurotrophic factor (MANF) protects against Aβ toxicity via attenuating Aβ-induced endoplasmic reticulum stress.","date":"2019","source":"Journal of neuroinflammation","url":"https://pubmed.ncbi.nlm.nih.gov/30760285","citation_count":71,"is_preprint":false},{"pmid":"25369767","id":"PMC_25369767","title":"Enhanced efficacy of the CDNF/MANF family by combined intranigral overexpression in the 6-OHDA rat model of Parkinson's disease.","date":"2014","source":"Molecular therapy : the journal of the American Society of Gene Therapy","url":"https://pubmed.ncbi.nlm.nih.gov/25369767","citation_count":68,"is_preprint":false},{"pmid":"29806020","id":"PMC_29806020","title":"Poststroke delivery of MANF promotes functional recovery in rats.","date":"2018","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/29806020","citation_count":67,"is_preprint":false},{"pmid":"20970425","id":"PMC_20970425","title":"Functions for the cardiomyokine, MANF, in cardioprotection, hypertrophy and heart failure.","date":"2010","source":"Journal of molecular and cellular cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/20970425","citation_count":66,"is_preprint":false},{"pmid":"26450777","id":"PMC_26450777","title":"Therapeutic potential of the endoplasmic reticulum located and secreted CDNF/MANF family of neurotrophic factors in Parkinson's disease.","date":"2015","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/26450777","citation_count":63,"is_preprint":false},{"pmid":"33299977","id":"PMC_33299977","title":"Neuroplastin Modulates Anti-inflammatory Effects of MANF.","date":"2020","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/33299977","citation_count":62,"is_preprint":false},{"pmid":"33856409","id":"PMC_33856409","title":"Feeding-induced hepatokine, Manf, ameliorates diet-induced obesity by promoting adipose browning via p38 MAPK pathway.","date":"2021","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33856409","citation_count":60,"is_preprint":false},{"pmid":"36739529","id":"PMC_36739529","title":"MANF regulates neuronal survival and UPR through its ER-located receptor IRE1α.","date":"2023","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/36739529","citation_count":58,"is_preprint":false},{"pmid":"29082311","id":"PMC_29082311","title":"MANF Is Essential for Neurite Extension and Neuronal Migration in the Developing Cortex.","date":"2017","source":"eNeuro","url":"https://pubmed.ncbi.nlm.nih.gov/29082311","citation_count":57,"is_preprint":false},{"pmid":"31781038","id":"PMC_31781038","title":"Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF) Is Highly Expressed in Mouse Tissues With Metabolic Function.","date":"2019","source":"Frontiers in endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/31781038","citation_count":56,"is_preprint":false},{"pmid":"33500254","id":"PMC_33500254","title":"Loss of MANF Causes Childhood-Onset Syndromic Diabetes Due to Increased Endoplasmic Reticulum Stress.","date":"2021","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/33500254","citation_count":55,"is_preprint":false},{"pmid":"36322628","id":"PMC_36322628","title":"Hormonal therapies up-regulate MANF and overcome female susceptibility to immune checkpoint inhibitor myocarditis.","date":"2022","source":"Science translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36322628","citation_count":54,"is_preprint":false},{"pmid":"32312869","id":"PMC_32312869","title":"MANF Promotes Diabetic Corneal Epithelial Wound Healing and Nerve Regeneration by Attenuating Hyperglycemia-Induced Endoplasmic Reticulum Stress.","date":"2020","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/32312869","citation_count":53,"is_preprint":false},{"pmid":"28924165","id":"PMC_28924165","title":"MANF regulates hypothalamic control of food intake and body weight.","date":"2017","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/28924165","citation_count":51,"is_preprint":false},{"pmid":"30305368","id":"PMC_30305368","title":"MANF Is Required for the Postnatal Expansion and Maintenance of Pancreatic β-Cell Mass in Mice.","date":"2018","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/30305368","citation_count":50,"is_preprint":false},{"pmid":"33460650","id":"PMC_33460650","title":"The cytoprotective protein MANF promotes neuronal survival independently from its role as a GRP78 cofactor.","date":"2021","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/33460650","citation_count":50,"is_preprint":false},{"pmid":"37838725","id":"PMC_37838725","title":"MANF stimulates autophagy and restores mitochondrial homeostasis to treat autosomal dominant tubulointerstitial kidney disease in mice.","date":"2023","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/37838725","citation_count":49,"is_preprint":false},{"pmid":"30967016","id":"PMC_30967016","title":"Armet, an aphid effector protein, induces pathogen resistance in plants by promoting the accumulation of salicylic acid.","date":"2019","source":"Philosophical transactions of the Royal Society of London. Series B, Biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/30967016","citation_count":48,"is_preprint":false},{"pmid":"29079145","id":"PMC_29079145","title":"Nrf2-mediated neuroprotection by MANF against 6-OHDA-induced cell damage via PI3K/AKT/GSK3β pathway.","date":"2017","source":"Experimental gerontology","url":"https://pubmed.ncbi.nlm.nih.gov/29079145","citation_count":48,"is_preprint":false},{"pmid":"29959908","id":"PMC_29959908","title":"MANF protects dopamine neurons and locomotion defects from a human α-synuclein induced Parkinson's disease model in C. elegans by regulating ER stress and autophagy pathways.","date":"2018","source":"Experimental neurology","url":"https://pubmed.ncbi.nlm.nih.gov/29959908","citation_count":48,"is_preprint":false},{"pmid":"32029702","id":"PMC_32029702","title":"Deficiency of the ER-stress-regulator MANF triggers progressive outer hair cell death and hearing loss.","date":"2020","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/32029702","citation_count":47,"is_preprint":false},{"pmid":"28698383","id":"PMC_28698383","title":"Pancreatic β-cell protection from inflammatory stress by the endoplasmic reticulum proteins thrombospondin 1 and mesencephalic astrocyte-derived neutrotrophic factor (MANF).","date":"2017","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/28698383","citation_count":45,"is_preprint":false},{"pmid":"33296727","id":"PMC_33296727","title":"MANF is neuroprotective against ethanol-induced neurodegeneration through ameliorating ER stress.","date":"2020","source":"Neurobiology of disease","url":"https://pubmed.ncbi.nlm.nih.gov/33296727","citation_count":44,"is_preprint":false},{"pmid":"32843919","id":"PMC_32843919","title":"Bone marrow mesenchymal stem cells induce M2 microglia polarization through PDGF-AA/MANF signaling.","date":"2020","source":"World journal of stem cells","url":"https://pubmed.ncbi.nlm.nih.gov/32843919","citation_count":43,"is_preprint":false},{"pmid":"29649564","id":"PMC_29649564","title":"XBP1 activation enhances MANF expression via binding to endoplasmic reticulum stress response elements within MANF promoter region in hepatitis B.","date":"2018","source":"The international journal of biochemistry & cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/29649564","citation_count":43,"is_preprint":false},{"pmid":"37799387","id":"PMC_37799387","title":"MANF brakes TLR4 signaling by competitively binding S100A8 with S100A9 to regulate macrophage phenotypes in hepatic fibrosis.","date":"2023","source":"Acta pharmaceutica Sinica. B","url":"https://pubmed.ncbi.nlm.nih.gov/37799387","citation_count":41,"is_preprint":false},{"pmid":"32845431","id":"PMC_32845431","title":"Trophic activities of endoplasmic reticulum proteins CDNF and MANF.","date":"2020","source":"Cell and tissue research","url":"https://pubmed.ncbi.nlm.nih.gov/32845431","citation_count":41,"is_preprint":false},{"pmid":"24587361","id":"PMC_24587361","title":"Spatiotemporal expression of MANF in the developing rat brain.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24587361","citation_count":41,"is_preprint":false},{"pmid":"29896089","id":"PMC_29896089","title":"Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF) Protects Against Neuronal Apoptosis via Activation of Akt/MDM2/p53 Signaling Pathway in a Rat Model of Intracerebral Hemorrhage.","date":"2018","source":"Frontiers in molecular neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/29896089","citation_count":40,"is_preprint":false},{"pmid":"36272696","id":"PMC_36272696","title":"Mesencephalic astrocyte-derived neurotrophic factor (MANF): Structure, functions and therapeutic potential.","date":"2022","source":"Ageing research reviews","url":"https://pubmed.ncbi.nlm.nih.gov/36272696","citation_count":38,"is_preprint":false},{"pmid":"24019940","id":"PMC_24019940","title":"Characterization of the structural and functional determinants of MANF/CDNF in Drosophila in vivo model.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24019940","citation_count":38,"is_preprint":false},{"pmid":"35135706","id":"PMC_35135706","title":"MANF: an emerging therapeutic target for metabolic diseases.","date":"2022","source":"Trends in endocrinology and metabolism: TEM","url":"https://pubmed.ncbi.nlm.nih.gov/35135706","citation_count":37,"is_preprint":false},{"pmid":"28719799","id":"PMC_28719799","title":"Mesencephalic astrocyte-derived neurotrophic factor (MANF), a new player in endoplasmic reticulum diseases: structure, biology, and therapeutic roles.","date":"2017","source":"Translational research : the journal of laboratory and clinical medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28719799","citation_count":36,"is_preprint":false},{"pmid":"33064897","id":"PMC_33064897","title":"Hepatocyte-derived MANF alleviates hepatic ischaemia-reperfusion injury via regulating endoplasmic reticulum stress-induced apoptosis in mice.","date":"2020","source":"Liver international : official journal of the International Association for the Study of the Liver","url":"https://pubmed.ncbi.nlm.nih.gov/33064897","citation_count":36,"is_preprint":false},{"pmid":"35238409","id":"PMC_35238409","title":"Armet from whitefly saliva acts as an effector to suppress plant defences by targeting tobacco cystatin.","date":"2022","source":"The New phytologist","url":"https://pubmed.ncbi.nlm.nih.gov/35238409","citation_count":35,"is_preprint":false},{"pmid":"32005751","id":"PMC_32005751","title":"MANF Ablation Causes Prolonged Activation of the UPR without Neurodegeneration in the Mouse Midbrain Dopamine System.","date":"2020","source":"eNeuro","url":"https://pubmed.ncbi.nlm.nih.gov/32005751","citation_count":35,"is_preprint":false},{"pmid":"33145627","id":"PMC_33145627","title":"Mono-macrophage-Derived MANF Protects Against Lipopolysaccharide-Induced Acute Kidney Injury via Inhibiting Inflammation and Renal M1 Macrophages.","date":"2020","source":"Inflammation","url":"https://pubmed.ncbi.nlm.nih.gov/33145627","citation_count":35,"is_preprint":false},{"pmid":"29887945","id":"PMC_29887945","title":"MANF improves the MPP+/MPTP-induced Parkinson's disease via improvement of mitochondrial function and inhibition of oxidative stress.","date":"2018","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/29887945","citation_count":35,"is_preprint":false},{"pmid":"26975047","id":"PMC_26975047","title":"Exploring the Conserved Role of MANF in the Unfolded Protein Response in Drosophila melanogaster.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26975047","citation_count":33,"is_preprint":false},{"pmid":"30147641","id":"PMC_30147641","title":"C. elegans MANF Homolog Is Necessary for the Protection of Dopaminergic Neurons and ER Unfolded Protein Response.","date":"2018","source":"Frontiers in neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/30147641","citation_count":33,"is_preprint":false},{"pmid":"36635421","id":"PMC_36635421","title":"MANF ameliorates DSS-induced mouse colitis via restricting Ly6ChiCX3CR1int macrophage transformation and suppressing CHOP-BATF2 signaling pathway.","date":"2023","source":"Acta pharmacologica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/36635421","citation_count":31,"is_preprint":false},{"pmid":"30515104","id":"PMC_30515104","title":"Trophic Factors in Inflammation and Regeneration: The Role of MANF and CDNF.","date":"2018","source":"Frontiers in physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30515104","citation_count":31,"is_preprint":false},{"pmid":"25511196","id":"PMC_25511196","title":"MANF silencing, immunity induction or autophagy trigger an unusual cell type in metamorphosing Drosophila brain.","date":"2014","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/25511196","citation_count":29,"is_preprint":false},{"pmid":"39147386","id":"PMC_39147386","title":"MANF facilitates breast cancer cell survival under glucose-starvation conditions via PRKN-mediated mitophagy regulation.","date":"2024","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/39147386","citation_count":28,"is_preprint":false},{"pmid":"32757264","id":"PMC_32757264","title":"DHA modulates MANF and TREM2 abundance, enhances neurogenesis, reduces infarct size, and improves neurological function after experimental ischemic stroke.","date":"2020","source":"CNS neuroscience & therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/32757264","citation_count":28,"is_preprint":false},{"pmid":"22243807","id":"PMC_22243807","title":"Valproic acid up-regulates melatonin MT1 and MT2 receptors and neurotrophic factors CDNF and MANF in the rat brain.","date":"2012","source":"The international journal of neuropsychopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/22243807","citation_count":28,"is_preprint":false},{"pmid":"35636177","id":"PMC_35636177","title":"Dendrobine inhibits dopaminergic neuron apoptosis via MANF-mediated ER stress suppression in MPTP/MPP+-induced Parkinson's disease models.","date":"2022","source":"Phytomedicine : international journal of phytotherapy and phytopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/35636177","citation_count":27,"is_preprint":false},{"pmid":"32275937","id":"PMC_32275937","title":"Liraglutide protects pancreatic β cells from endoplasmic reticulum stress by upregulating MANF to promote autophagy turnover.","date":"2020","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32275937","citation_count":27,"is_preprint":false},{"pmid":"37118549","id":"PMC_37118549","title":"Aging disrupts MANF-mediated immune modulation during skeletal muscle regeneration.","date":"2023","source":"Nature aging","url":"https://pubmed.ncbi.nlm.nih.gov/37118549","citation_count":26,"is_preprint":false},{"pmid":"35129674","id":"PMC_35129674","title":"CDNF and MANF regulate ER stress in a tissue-specific manner.","date":"2022","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/35129674","citation_count":25,"is_preprint":false},{"pmid":"30386256","id":"PMC_30386256","title":"Emerging Roles for Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF) in Pancreatic Beta Cells and Diabetes.","date":"2018","source":"Frontiers in physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30386256","citation_count":25,"is_preprint":false},{"pmid":"29687079","id":"PMC_29687079","title":"Photoreceptor Protection by Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF).","date":"2018","source":"eNeuro","url":"https://pubmed.ncbi.nlm.nih.gov/29687079","citation_count":25,"is_preprint":false},{"pmid":"20214902","id":"PMC_20214902","title":"Solution structure and dynamics of mouse ARMET.","date":"2010","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/20214902","citation_count":24,"is_preprint":false},{"pmid":"33127565","id":"PMC_33127565","title":"Hepatocyte-derived MANF is protective for rifampicin-induced cholestatic hepatic injury via inhibiting ATF4-CHOP signal activation.","date":"2020","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33127565","citation_count":24,"is_preprint":false},{"pmid":"33071755","id":"PMC_33071755","title":"Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF) Regulates Neurite Outgrowth Through the Activation of Akt/mTOR and Erk/mTOR Signaling Pathways.","date":"2020","source":"Frontiers in molecular neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/33071755","citation_count":24,"is_preprint":false},{"pmid":"26820513","id":"PMC_26820513","title":"A Comparative Analysis of the Molecular Features of MANF and CDNF.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26820513","citation_count":23,"is_preprint":false},{"pmid":"29797541","id":"PMC_29797541","title":"MANF attenuates neuronal apoptosis and promotes behavioral recovery via Akt/MDM-2/p53 pathway after traumatic spinal cord injury in rats.","date":"2018","source":"BioFactors (Oxford, England)","url":"https://pubmed.ncbi.nlm.nih.gov/29797541","citation_count":23,"is_preprint":false},{"pmid":"32229226","id":"PMC_32229226","title":"Molecular profile of the rat peri-infarct region four days after stroke: Study with MANF.","date":"2020","source":"Experimental neurology","url":"https://pubmed.ncbi.nlm.nih.gov/32229226","citation_count":23,"is_preprint":false},{"pmid":"27608005","id":"PMC_27608005","title":"MRI Dynamically Evaluates the Therapeutic Effect of Recombinant Human MANF on Ischemia/Reperfusion Injury in Rats.","date":"2016","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/27608005","citation_count":22,"is_preprint":false},{"pmid":"34745419","id":"PMC_34745419","title":"MANF: A Novel Endoplasmic Reticulum Stress Response Protein-The Role in Neurological and Metabolic Disorders.","date":"2021","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/34745419","citation_count":21,"is_preprint":false},{"pmid":"36436758","id":"PMC_36436758","title":"Mesencephalic astrocyte-derived neurotrophic factor (MANF) prevents the neuroinflammation induced dopaminergic neurodegeneration.","date":"2022","source":"Experimental gerontology","url":"https://pubmed.ncbi.nlm.nih.gov/36436758","citation_count":21,"is_preprint":false},{"pmid":"35911675","id":"PMC_35911675","title":"Cold Plasma Irradiation Attenuates Atopic Dermatitis via Enhancing HIF-1α-Induced MANF Transcription Expression.","date":"2022","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/35911675","citation_count":21,"is_preprint":false},{"pmid":"37048105","id":"PMC_37048105","title":"Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF): An Emerging Therapeutic Target for Neurodegenerative Disorders.","date":"2023","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/37048105","citation_count":20,"is_preprint":false},{"pmid":"35972224","id":"PMC_35972224","title":"MANF in POMC Neurons Promotes Brown Adipose Tissue Thermogenesis and Protects Against Diet-Induced Obesity.","date":"2022","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/35972224","citation_count":20,"is_preprint":false},{"pmid":"38134597","id":"PMC_38134597","title":"Mesencephalic astrocyte-derived neurotrophic factor (MANF) alleviates cerebral ischemia/reperfusion injury in mice by regulating microglia polarization via A20/NF-κB pathway.","date":"2023","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/38134597","citation_count":20,"is_preprint":false},{"pmid":"36585419","id":"PMC_36585419","title":"MANF/EWSR1/ANXA6 pathway might as the bridge between hypolipidemia and major depressive disorder.","date":"2022","source":"Translational psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/36585419","citation_count":19,"is_preprint":false},{"pmid":"26429332","id":"PMC_26429332","title":"ER stress-inducible protein MANF selectively expresses in human spleen.","date":"2015","source":"Human immunology","url":"https://pubmed.ncbi.nlm.nih.gov/26429332","citation_count":19,"is_preprint":false},{"pmid":"35783104","id":"PMC_35783104","title":"UPR Responsive Genes Manf and Xbp1 in Stroke.","date":"2022","source":"Frontiers in cellular neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/35783104","citation_count":18,"is_preprint":false},{"pmid":"24845376","id":"PMC_24845376","title":"A sensitive assay for the biosynthesis and secretion of MANF using NanoLuc activity.","date":"2014","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/24845376","citation_count":18,"is_preprint":false},{"pmid":"33994992","id":"PMC_33994992","title":"Increased MANF Expression in the Inferior Temporal Gyrus in Patients With Alzheimer Disease.","date":"2021","source":"Frontiers in aging neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/33994992","citation_count":17,"is_preprint":false},{"pmid":"33644980","id":"PMC_33644980","title":"MANF protects pancreatic acinar cells against alcohol-induced endoplasmic reticulum stress and cellular injury.","date":"2021","source":"Journal of hepato-biliary-pancreatic sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33644980","citation_count":17,"is_preprint":false},{"pmid":"31138438","id":"PMC_31138438","title":"MANF deletion abrogates early larval Caenorhabditis elegans stress response to tunicamycin and Pseudomonas aeruginosa.","date":"2019","source":"European journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/31138438","citation_count":17,"is_preprint":false},{"pmid":"25917377","id":"PMC_25917377","title":"Analysis of the conserved neurotrophic factor MANF in the Drosophila adult brain.","date":"2015","source":"Gene expression patterns : GEP","url":"https://pubmed.ncbi.nlm.nih.gov/25917377","citation_count":16,"is_preprint":false},{"pmid":"30555354","id":"PMC_30555354","title":"MANF: A New Player in the Control of Energy Homeostasis, and Beyond.","date":"2018","source":"Frontiers in physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30555354","citation_count":15,"is_preprint":false},{"pmid":"34305531","id":"PMC_34305531","title":"MANF Is Neuroprotective in Early Stages of EAE, and Elevated in Spinal White Matter by Treatment With Dexamethasone.","date":"2021","source":"Frontiers in cellular neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/34305531","citation_count":15,"is_preprint":false},{"pmid":"37555005","id":"PMC_37555005","title":"CDNF and MANF in the brain dopamine system and their potential as treatment for Parkinson's disease.","date":"2023","source":"Frontiers in psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/37555005","citation_count":14,"is_preprint":false},{"pmid":"32119994","id":"PMC_32119994","title":"MANF delivery improves retinal homeostasis and cell replacement therapies in ageing mice.","date":"2020","source":"Experimental gerontology","url":"https://pubmed.ncbi.nlm.nih.gov/32119994","citation_count":14,"is_preprint":false},{"pmid":"25947918","id":"PMC_25947918","title":"Characterization of the Role of MANF in Regulating the Secretion of CRELD2.","date":"2015","source":"Biological & pharmaceutical bulletin","url":"https://pubmed.ncbi.nlm.nih.gov/25947918","citation_count":14,"is_preprint":false},{"pmid":"31037983","id":"PMC_31037983","title":"Secondary Pathology of the Thalamus after Focal Cortical Stroke in Rats is not Associated with Thermal or Mechanical Hypersensitivity and is Not Alleviated by Intra-Thalamic Post-Stroke Delivery of Recombinant CDNF or MANF.","date":"2019","source":"Cell transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/31037983","citation_count":14,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":49681,"output_tokens":8932,"usd":0.141512,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":18931,"output_tokens":4515,"usd":0.103765,"stage2_stop_reason":"end_turn"},"total_usd":0.245277,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2003,\n      \"finding\": \"MANF (initially derived from a rat mesencephalic type-1 astrocyte cell line) is a secreted ~20 kDa protein that selectively protects nigral dopaminergic neurons in vitro, with greater selectivity than GDNF or BDNF at low-to-middle concentrations; contains two conserved domains and eight cysteines conserved from C. elegans to human.\",\n      \"method\": \"Recombinant protein production, in vitro neuronal survival assays comparing MANF, GDNF, and BDNF dose-response on dopaminergic vs. GABAergic/serotonergic neurons\",\n      \"journal\": \"Journal of molecular neuroscience : MN\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro functional assay with concentration-response, recombinant protein, but single lab and limited mechanistic depth\",\n      \"pmids\": [\"12794311\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Mouse ARMET/MANF is an 18 kDa soluble ER protein that, after signal-sequence cleavage, contains four intramolecular disulfide bonds including two in CXXC sequences; its UPR-dependent transcriptional induction is regulated by an ERSE-II element (identical to the HERP gene ERSE-II) in the MANF promoter.\",\n      \"method\": \"Western blotting, in vitro disulfide-bond characterization, reporter gene assay with MANF promoter deletion constructs\",\n      \"journal\": \"Cell structure and function\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — biochemical characterization of protein maturation (disulfide bonds) combined with reporter gene assay for promoter element; multiple orthogonal methods in one study\",\n      \"pmids\": [\"17507765\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"ARMET/MANF protein is upregulated by multiple forms of ER stress in several cell lines and by cerebral ischemia in rat; it localizes to the ER and Golgi and is also secreted; siRNA-mediated silencing increases susceptibility to ER stress-induced death and paradoxically increases proliferation, while overexpression inhibits proliferation and improves viability under stress, establishing MANF as a secreted mediator of the adaptive UPR pathway.\",\n      \"method\": \"siRNA knockdown, overexpression, immunofluorescence (ER/Golgi localization), cell viability assays, immunohistochemistry in rat ischemia model\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal gain- and loss-of-function with defined phenotypic readouts, direct localization, multiple cell lines and an in vivo model\",\n      \"pmids\": [\"18561914\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Crystal structures of human MANF and CDNF reveal that neither resembles known growth factors; the N-terminal domain is a saposin-like lipid-binding domain (suggesting membrane/lipid interaction), while the natively unfolded C-terminal domain of MANF contains a CKGC disulfide bridge analogous to reductases/disulfide isomerases, consistent with a role in ER stress response. Three residues (I10, E79, K88 in MANF) may account for functional differences between MANF and CDNF.\",\n      \"method\": \"X-ray crystallography (crystal structures of both proteins solved), structural bioinformatics\",\n      \"journal\": \"Protein engineering, design & selection : PEDS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structures solved and validated; structural basis for bifunctionality established; independently consistent with NMR study (PMID 21047780)\",\n      \"pmids\": [\"19258449\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Solution NMR structure of human MANF shows the C-terminal domain (C-MANF) is homologous to the SAP domain of Ku70, a known inhibitor of pro-apoptotic Bax; cellular experiments confirm that both full-length MANF and C-MANF protect neurons intracellularly as efficiently as Ku70.\",\n      \"method\": \"NMR structure determination, cellular neuroprotection assays with MANF and C-MANF constructs\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — NMR structure with functional validation in cells; structural homology to Ku70 SAP domain experimentally supported by cellular assays\",\n      \"pmids\": [\"21047780\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Solution structure of mouse ARMET/MANF (by NMR relaxation and trypsin digestion) reveals an entirely α-helical two-domain architecture in which the N-terminal and C-terminal domains tumble independently; positive charges are dispersed across both domains and the linker, suggesting a negatively-charged ligand-binding mode.\",\n      \"method\": \"NMR solution structure, 15N relaxation experiments, trypsin digestion\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — NMR structure with dynamics characterization; independent of but consistent with crystal structure (PMID 19258449)\",\n      \"pmids\": [\"20214902\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The C-terminal RTDL sequence of MANF is required for both ER retention and stress-responsive secretion; KDEL receptor (KDELR) overexpression reduces MANF secretion only when RTDL is present; MANF binds to KDELRs at the plasma membrane surface (inhibited by a KDELR-interacting peptide), and surface KDELR levels increase after ER stress.\",\n      \"method\": \"Lentiviral vector constructs with/without RTDL, KDELR overexpression, peptide competition, immunofluorescence, neuroblastoma cells and primary cortical neurons\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal approaches (deletion mutants, overexpression, peptide competition, FACS/imaging); two cell systems\",\n      \"pmids\": [\"23255601\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"In Drosophila, only full-length MANF containing both N-terminal saposin-like and C-terminal SAP domains rescues larval lethality of DmManf null mutants; neither domain alone (even co-expressed) is sufficient; deleting the signal peptide or mutating the CXXC motif in the C-terminal domain destroys activity; positively charged surface residues and the C-terminal ER retention signal are necessary for rescue when expression is restricted.\",\n      \"method\": \"Transgenic rescue experiments in DmManf mutant background, domain-deletion and point-mutation constructs, in vitro sympathetic neuron survival assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vivo genetic rescue with multiple structure–function mutants; functional conservation confirmed in mammalian neuronal assay\",\n      \"pmids\": [\"24019940\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MANF is indispensable for pancreatic β-cell proliferation and survival in mice: MANF-deficient mice develop progressive postnatal loss of β-cell mass causing diabetes, with chronic UPR activation in islets; recombinant MANF enhances β-cell proliferation in vitro; MANF overexpression in diabetic mouse pancreas enhances β-cell regeneration.\",\n      \"method\": \"MANF knockout mice (loss-of-function), in vitro proliferation assays with recombinant MANF, in vivo pancreatic MANF overexpression in diabetic mice, UPR marker analysis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal loss- and gain-of-function in vivo and in vitro with specific phenotypic readouts; replicated across models\",\n      \"pmids\": [\"24726366\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Reduced chaperone (Hsc70) activity with aging leads to decreased association of mutant TBP with XBP1s, reducing XBP1s-driven transcription of MANF; overexpression of MANF ameliorates Purkinje cell degeneration via PKC-dependent signaling; Hsc70 overexpression restores TBP-XBP1s interaction and MANF transcription.\",\n      \"method\": \"SCA17 knockin mice with tamoxifen-inducible expression, co-IP (TBP-XBP1s interaction), Hsc70 overexpression, PKC inhibitor studies, viral MANF overexpression\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis in vivo, Co-IP for interaction, pharmacological pathway dissection, multiple orthogonal methods\",\n      \"pmids\": [\"24462098\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"PDGF-like signaling induces MANF expression in innate immune cells of damaged retina (in both flies and mice); MANF promotes alternative (M2) activation of innate immune cells, enhancing neuroprotection and tissue repair, and improves photoreceptor replacement therapy success.\",\n      \"method\": \"Genetic epistasis in Drosophila retina damage model, mouse retinal damage model, MANF gain/loss-of-function, immune cell phenotyping, photoreceptor transplantation assay\",\n      \"journal\": \"Science (New York, N.Y.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conserved pathway established in two organisms (fly and mouse) with genetic and pharmacological tools; multiple phenotypic readouts\",\n      \"pmids\": [\"27365452\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In Drosophila, Manf shows genetic interactions with GRP78 (Hsc70-3), PERK (PEK), and XBP1 homologs, placing DmManf as a regulator of the UPR; DmManf expression is upregulated by pharmacological ER stress inducers, a response conserved between Drosophila and mammals.\",\n      \"method\": \"Genetic interaction assays (double mutant/RNAi combinations), drug-induced ER stress, reporter gene expression\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis in Drosophila; conservation supported by parallel mammalian data; single lab\",\n      \"pmids\": [\"26975047\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"MANF is highly enriched in hypothalamic nuclei and its expression is upregulated by fasting; increasing hypothalamic MANF promotes hyperphagia, decreasing it causes hypophagia; mechanistically, MANF enhances ER localization and activity of PIP4k2b kinase, thereby triggering hypothalamic insulin resistance.\",\n      \"method\": \"Viral vector-mediated overexpression and knockdown in mouse hypothalamus, food intake/body weight measurements, PIP4k2b co-localization and activity assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo gain- and loss-of-function with specific signaling pathway dissection; single lab but multiple approaches\",\n      \"pmids\": [\"28924165\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"MANF-deficient neural stem cells show no proliferation defect but exhibit impaired neurite extension upon differentiation; in vivo MANF deletion causes slower neuronal migration and impaired neurite outgrowth, preceded by reduced de novo protein synthesis and constitutively activated UPR pathways.\",\n      \"method\": \"MANF knockout NSC cultures, in utero electroporation/cortical migration assay, neurite outgrowth measurement, protein synthesis assay, UPR pathway marker analysis\",\n      \"journal\": \"eNeuro\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo and in vitro loss-of-function with mechanistic UPR pathway readouts; multiple orthogonal methods\",\n      \"pmids\": [\"29082311\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"MANF promotes differentiation and migration of neural progenitor cells (NPCs) in vitro through activation of STAT3 and ERK1/2; in SVZ explants, MANF overexpression facilitates cell migration via STAT3 and ERK1/2; in vivo, intracerebroventricular MANF promotes migration of DCX+ neuroblasts after stroke.\",\n      \"method\": \"NSC/NPC culture with recombinant MANF, SVZ explant migration assay, STAT3/ERK phosphorylation Western blot, rat cortical stroke model with ICV injection\",\n      \"journal\": \"Molecular therapy : the journal of the American Society of Gene Therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro signaling assays plus in vivo model; pathway identified by Western blot; single lab\",\n      \"pmids\": [\"29050872\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"In human β-cells, exogenous recombinant MANF reduces cytokine-induced cell death by ~38%; the protective effect is associated with repression of NF-κB signaling and amelioration of ER stress; MANF knockdown aggravates ER stress after cytokine challenge.\",\n      \"method\": \"Primary human islets + recombinant MANF, siRNA knockdown in EndoC-βH1 cells, cell viability, global transcriptomics, NF-κB pathway analysis\",\n      \"journal\": \"Diabetologia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal gain/loss-of-function in human cells with defined pathway readout; single lab\",\n      \"pmids\": [\"30032427\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The SAP domain of MANF selectively associates with the nucleotide-binding domain (NBD) of ADP-bound BiP; crystal structures show the SAP domain engages the cleft between NBD subdomains Ia and IIa, stabilizing ADP-bound conformation and clashing with the ATP-bound interdomain linker; MANF inhibits both ADP release from and ATP binding to BiP, thereby inhibiting client release; cells lacking MANF have fewer ER stress-induced BiP-containing high-molecular-weight complexes.\",\n      \"method\": \"Crystal structures of MANF SAP domain–BiP NBD complex, in vitro nucleotide exchange assays, MANF knockout cells, size-exclusion chromatography of BiP complexes\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus in vitro enzymatic assay plus cell-based validation; multiple orthogonal methods; single lab but rigorous\",\n      \"pmids\": [\"30710085\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MANF deficiency in flies causes enhanced inflammation and shorter lifespan; MANF deficiency in mice causes inflammatory phenotypes and progressive liver damage (fibrosis, steatosis); immune cell-derived MANF specifically protects against liver inflammation and fibrosis, while hepatocyte-derived MANF prevents hepatosteatosis; liver rejuvenation by heterochronic parabiosis depends on MANF.\",\n      \"method\": \"Drosophila MANF overexpression/KO, MANF heterozygous mice, tissue-specific MANF deletion (immune cells vs. hepatocytes), heterochronic parabiosis, recombinant MANF supplementation\",\n      \"journal\": \"Nature metabolism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cell-type-specific KOs, parabiosis, recombinant protein rescue, conserved across two organisms; multiple orthogonal methods\",\n      \"pmids\": [\"31489403\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Brain-specific MANF deletion exacerbates ethanol-induced neuronal apoptosis and ER stress; blocking ER stress abrogates the detrimental effects of MANF deficiency on ethanol-induced apoptosis, placing MANF upstream of ER stress in ethanol neurotoxicity; in a tunicamycin-induced neurodegeneration model, MANF deficiency potentiates ER stress and neurodegeneration.\",\n      \"method\": \"CNS-specific Manf knockout mice, tunicamycin and ethanol in vivo models, ER stress marker analysis, RNA-seq\",\n      \"journal\": \"Neurobiology of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with pharmacological epistasis (ER stress inhibitor rescue); pathway positioning established; single lab\",\n      \"pmids\": [\"33296727\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"MANF loss causes prolonged activation of the IRE1α branch of the UPR (and in aged mice additionally PERK and ATF6 branches) without causing dopaminergic neuron loss in the substantia nigra or behavioral changes; cortical neurons lacking MANF are more vulnerable to additional ER stress in vitro, indicating endogenous MANF is required for neuronal ER homeostasis maintenance.\",\n      \"method\": \"Brain-specific MANF knockout mice, UPR marker analysis at multiple ages, stereological neuron counting in SN, dopamine measurement, cortical neuron cultures with ER stress challenge\",\n      \"journal\": \"eNeuro\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with comprehensive pathway and phenotypic analysis in vivo and in vitro; multiple UPR branches characterized\",\n      \"pmids\": [\"32005751\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Neuroplastin (NPTN) is a cell-surface receptor for MANF; biochemical analysis shows physiological interaction between MANF and NPTN on the cell surface; MANF binding to NPTN suppresses NF-κB signaling, mitigating inflammatory response and apoptosis.\",\n      \"method\": \"Co-immunoprecipitation, surface binding assays, NPTN knockdown/overexpression, NF-κB reporter assays\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Co-IP and surface binding with functional NF-κB readout; single lab; mechanistic pathway proposed with supporting experiments\",\n      \"pmids\": [\"33299977\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"MANF regulates neurite outgrowth by activating Akt/mTOR and Erk/mTOR signaling pathways; MANF KO N2a cells fail to grow neurites on RA stimulation, and this is rescued by MANF overexpression or exogenous MANF; pharmacological blockade of Akt, Erk, or mTOR eliminates the MANF-promoting effect on neurite outgrowth.\",\n      \"method\": \"CRISPR/Cas9 MANF KO in N2a cells, siRNA in SH-SY5Y, adenoviral MANF overexpression, pharmacological inhibitors of Akt/Erk/mTOR, Western blot for pathway activation\",\n      \"journal\": \"Frontiers in molecular neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR KO, siRNA, overexpression, and pharmacological epistasis; multiple cell lines; single lab\",\n      \"pmids\": [\"33071755\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MANF promotes neuronal survival as a general UPR regulator affecting multiple UPR pathways simultaneously, independently of its role as a GRP78 cofactor; MANF interacts with a conserved set of 15 ER proteins including GRP78, GRP170, PDIA1, and PDIA6; ATP binding to MANF (confirmed by MST and NMR) blocks the MANF-GRP78 interaction; antiapoptotic activity of MANF mutants in neurons reveals divergent roles as GRP78 cofactor vs. antiapoptotic UPR regulator.\",\n      \"method\": \"AP-MS interactome from two mammalian cell lines, microscale thermophoresis (ATP binding), NMR, neuronal apoptosis assays with MANF mutants\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — biochemical ATP-binding assay (MST + NMR) plus interactome plus neuronal functional assays; multiple orthogonal methods; single lab\",\n      \"pmids\": [\"33460650\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Loss-of-function mutations in the MANF gene in humans cause childhood-onset diabetes with neurodevelopmental disorder; MANF knockout in human embryonic stem cell-derived β-cells induces mild ER stress and impairs insulin-processing capacity; upon implantation in diabetic mice, MANF-KO grafts show elevated ER stress and functional failure.\",\n      \"method\": \"Human genetic (homozygous LOF mutations), hESC MANF knockout, β-cell differentiation, insulin processing assay, xenotransplantation in immunocompromised diabetic mice\",\n      \"journal\": \"Diabetes\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — human genetics validated mechanistically with hESC KO and in vivo xenograft; multiple orthogonal approaches\",\n      \"pmids\": [\"33500254\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"XBP1s (spliced XBP1) binds to ERSE-I in the MANF promoter to promote MANF transcription; MANF in turn interacts with XBP1s to enhance its own expression in a positive feedback loop; IRE1α endonuclease inhibition or XBP1s knockdown attenuates MANF expression.\",\n      \"method\": \"ChIP, promoter reporter assay, XBP1s overexpression and siRNA knockdown, IRE1α inhibitor, co-IP of MANF with XBP1s\",\n      \"journal\": \"The international journal of biochemistry & cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and reporter gene assay plus Co-IP; multiple methods; single lab\",\n      \"pmids\": [\"29649564\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Liver-specific MANF overexpression protects against high-fat diet-induced obesity and promotes browning (beige adipogenesis) of inguinal WAT; mechanistically, MANF directly promotes white adipocyte browning via the p38 MAPK pathway; p38 MAPK blockade abolishes MANF-induced browning; liver-specific MANF KO impairs WAT browning and worsens diet-induced obesity.\",\n      \"method\": \"Liver-specific overexpression and KO mouse models, p38 MAPK inhibitor, primary white adipocyte culture with recombinant MANF, thermogenesis markers\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal liver-specific OE and KO in vivo, pharmacological epistasis, in vitro mechanistic confirmation; multiple orthogonal approaches\",\n      \"pmids\": [\"33856409\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MANF is transcriptionally induced in cardiomyocytes by liganded estrogen receptor β and inhibited by androgen receptor; ICI (immune checkpoint inhibitor) treatment reduces serum estradiol, decreasing cardiac MANF/HSPA5; heart-specific MANF deletion amplifies ICI myocarditis, and recombinant MANF protein attenuates it, establishing a causal role.\",\n      \"method\": \"Genetic cardiac-specific Manf deletion, recombinant MANF administration, ERβ agonist and androgen depletion in vivo, transcriptional regulation analysis\",\n      \"journal\": \"Science translational medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO, recombinant protein rescue, and pharmacological pathway dissection; multiple orthogonal methods in one study\",\n      \"pmids\": [\"36322628\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MANF directly interacts with the ER transmembrane UPR sensor IRE1α; wild-type MANF (but not its IRE1α-binding-deficient mutant) decreases IRE1α oligomerization and phosphorylation, reduces XBP1 splicing, ATF6 and TXNIP levels, and protects neurons from ER stress-induced death; MANF-IRE1α interaction (not MANF-BiP interaction) is required for protection of dopamine neurons in a Parkinson's disease animal model.\",\n      \"method\": \"Co-IP of MANF with IRE1α, binding-interface mutagenesis, IRE1α oligomerization/phosphorylation assays, XBP1 splicing reporter, neuronal apoptosis assays, 6-OHDA rat PD model with MANF mutant constructs\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — direct protein interaction mapped to binding interface, multiple UPR functional readouts, mutant dissection of MANF-IRE1α vs. MANF-BiP, in vivo PD model validation\",\n      \"pmids\": [\"36739529\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MANF overexpression in kidney tubular cells stimulates autophagy/mitophagy and mitochondrial biogenesis through p-AMPK enhancement, clears mutant UMOD aggregates, and suppresses cGAS-STING activation; genetic ablation of tubular MANF worsens autophagy suppression and kidney fibrosis in a ADTKD-UMOD mouse model.\",\n      \"method\": \"Inducible tubular MANF overexpression and knockout in ADTKD-UMOD mice, autophagy/mitophagy markers, AMPK phosphorylation, cGAS-STING pathway analysis, mutant UMOD clearance\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal in vivo gain- and loss-of-function with mechanistic pathway identification (AMPK, autophagy, cGAS-STING); disease-relevant model\",\n      \"pmids\": [\"37838725\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MANF brakes macrophage TLR4 signaling by competitively binding S100A8, preventing S100A8/A9 heterodimer formation and thereby inhibiting S100A8/A9-mediated TLR4-NF-κB activation; myeloid-specific MANF KO increases hepatic Ly6Chigh pro-inflammatory macrophages and worsens hepatic fibrosis.\",\n      \"method\": \"Myeloid-specific MANF KO mice, competitive binding assay (MANF vs. S100A9 for S100A8), Co-IP, macrophage phenotyping, TLR4-NF-κB pathway analysis, recombinant MANF treatment\",\n      \"journal\": \"Acta pharmaceutica Sinica. B\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO plus direct competitive binding assay plus signaling pathway validation; multiple orthogonal methods\",\n      \"pmids\": [\"37799387\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Under glucose starvation, SENP1-mediated de-SUMOylation of MANF inhibits its nuclear translocation and increases cytoplasmic/mitochondrial MANF; mitochondrial MANF binds to PRKN (Parkin) E3 ligase, alleviates oxidative inhibition of PRKN's RING II domain via its CXXC motif, restores PRKN E3 ligase activity, and thereby mediates mitophagy to promote breast cancer cell survival.\",\n      \"method\": \"SENP1 manipulation (de-SUMOylation), Co-IP of MANF-PRKN, PRKN ubiquitin ligase activity assay, CXXC motif mutagenesis, mitophagy assays, MANF subcellular fractionation\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, enzymatic activity restoration, mutagenesis of CXXC motif; multiple methods; single lab\",\n      \"pmids\": [\"39147386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"ARMET/MANF interacts with mutant matrilin-3 (but not COMP) in chondrocyte ER stress models of skeletal dysplasia, establishing substrate specificity; both ARMET and CRELD2 are secreted into ECM following ER stress; substrate-trapping experiments confirmed CRELD2 but not ARMET has PDI-like activity.\",\n      \"method\": \"Cell and mouse models of chondrodysplasia, co-IP/substrate trapping, immunohistochemistry, Western blotting; genotype-specific expression analysis\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — substrate specificity shown by Co-IP and substrate trapping; genotype-specific interaction validated; single lab\",\n      \"pmids\": [\"23956175\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"THBS1 (thrombospondin 1) maintains MANF expression in β-cells exposed to cytokines or thapsigargin, and this THBS1-MANF axis prevents BIM-dependent mitochondrial apoptosis; prolonged stress leads to THBS1 and MANF degradation and loss of this prosurvival mechanism.\",\n      \"method\": \"THBS1 overexpression/silencing in rat/mouse/human β-cells, Western blot for MANF and BIM, apoptosis assays, ER stress induction\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pathway epistasis (THBS1→MANF→BIM) with gain/loss-of-function and specific apoptotic readout; multiple species tested; single lab\",\n      \"pmids\": [\"28698383\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CDNF and MANF display functional redundancy in skeletal muscle (CDNF deficiency increases UPR activation, aggravated by combined MANF loss) but not in brain; neither single nor combined brain-specific KO causes dopaminergic neuron degeneration, showing tissue-specific UPR regulation.\",\n      \"method\": \"Cdnf−/−, Manf−/−, and double KO mice, UPR markers in multiple tissues, dopaminergic neuron counting\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — double KO with tissue-specific phenotypic analysis; defines functional redundancy/divergence; single lab\",\n      \"pmids\": [\"35129674\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MANF is a bifunctional ER-resident/secreted protein whose N-terminal saposin-like domain mediates lipid/membrane interaction and extracellular neurotrophic activity, while its natively unfolded C-terminal SAP/CXXC domain acts intracellularly as a nucleotide-exchange inhibitor of the Hsp70 chaperone BiP (stabilizing ADP-bound BiP–client complexes) and as a direct binding partner and negative regulator of the UPR sensor IRE1α (reducing its oligomerization, phosphorylation, and XBP1 splicing); extracellularly, MANF binds the cell-surface receptor Neuroplastin to suppress NF-κB-mediated inflammation, competes with S100A9 for S100A8 to brake TLR4–NF-κB macrophage activation, and is retained in the ER via its C-terminal RTDL sequence through interaction with KDEL receptors whose surface levels rise after ER stress; loss of MANF in mice causes chronic UPR activation, progressive β-cell death and diabetes, impaired neurite outgrowth, and tissue-specific pathologies (liver fibrosis, outer hair cell death, hearing loss), while its protective signaling engages PI3K/Akt/mTOR, ERK/mTOR, Nrf2, and AMPK pathways depending on context.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MANF is a bifunctional ER-resident and secreted protein that maintains endoplasmic reticulum proteostasis and confers cytoprotection across neuronal, pancreatic, hepatic, immune, and cardiac tissues, functioning as a secreted mediator of the adaptive unfolded protein response (UPR) [#2, #8]. Its two-domain architecture underlies this bifunctionality: an N-terminal saposin-like lipid-binding domain and a natively unfolded C-terminal SAP/CXXC domain homologous to the Bax inhibitor Ku70, with both domains required for full activity in vivo [#3, #4, #7]. Intracellularly, the SAP domain binds the nucleotide-binding domain of ADP-bound BiP, stabilizing the ADP conformation and inhibiting client release, and ATP binding to MANF disrupts this interaction [#16, #22]; MANF also directly binds the UPR sensor IRE1\\u03b1, reducing its oligomerization and phosphorylation and dampening XBP1 splicing, and this IRE1\\u03b1 interaction \\u2014 distinct from BiP binding \\u2014 is required for dopaminergic neuroprotection [#27]. MANF expression is induced by ER stress through ERSE elements and an XBP1s-driven positive feedback loop [#1, #24]. The C-terminal RTDL signal mediates ER retention and stress-responsive secretion via KDEL receptors [#6]. Extracellularly, MANF acts through the cell-surface receptor Neuroplastin to suppress NF-\\u03baB signaling and competes with S100A9 for S100A8 to brake TLR4\\u2013NF-\\u03baB macrophage activation, promoting anti-inflammatory immune phenotypes and tissue repair [#10, #20, #29], and engages PI3K/Akt/mTOR, ERK/mTOR, STAT3, AMPK, and p38 MAPK signaling in a context-dependent manner to promote neurite outgrowth, autophagy, and metabolic regulation [#14, #21, #28, #25]. Loss of MANF causes chronic UPR activation with progressive \\u03b2-cell death and diabetes, impaired neurite outgrowth and neuronal migration, and tissue-specific liver and inflammatory pathology [#8, #13, #17]. Homozygous loss-of-function MANF mutations cause childhood-onset diabetes with a neurodevelopmental disorder [#23].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established MANF as a secreted factor with selective trophic activity, defining the founding functional question of how a non-canonical protein protects specific neuron populations.\",\n      \"evidence\": \"Recombinant protein with in vitro dopaminergic neuron survival assays comparing MANF, GDNF, and BDNF\",\n      \"pmids\": [\"12794311\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No receptor or molecular mechanism for selectivity identified\", \"Single cell-line-derived source\", \"No structural basis for activity\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Positioned MANF within the UPR by showing ER stress induces it and its loss/gain reciprocally modulates stress survival, recasting the trophic factor as a UPR effector.\",\n      \"evidence\": \"siRNA knockdown, overexpression, ER/Golgi immunofluorescence, viability assays, and rat ischemia model\",\n      \"pmids\": [\"18561914\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular target within the UPR unknown\", \"Mechanism of cytoprotection not defined\", \"Secreted vs. intracellular roles not separated\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Resolved the structural basis of MANF's bifunctionality, distinguishing a lipid-binding N-domain from a Ku70-like SAP/CXXC C-domain implicated in apoptosis suppression and redox activity.\",\n      \"evidence\": \"X-ray crystallography and NMR solution structures with cellular neuroprotection assays of full-length and C-terminal constructs\",\n      \"pmids\": [\"19258449\", \"21047780\", \"20214902\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No binding partner for either domain identified\", \"Mechanism of SAP-domain neuroprotection unresolved\", \"Functional ligand of the saposin-like domain unknown\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined how MANF balances ER retention against secretion, showing the RTDL signal and KDEL receptors gate stress-responsive release.\",\n      \"evidence\": \"RTDL deletion constructs, KDELR overexpression, peptide competition, and imaging in neuroblastoma and primary cortical neurons\",\n      \"pmids\": [\"23255601\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Surface KDELR-MANF signaling consequences not defined\", \"Trigger linking ER stress to surface KDELR increase unclear\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Demonstrated through in vivo genetic rescue that both MANF domains are jointly required, establishing the protein acts as an integrated bifunctional unit rather than two separable activities.\",\n      \"evidence\": \"Transgenic rescue of DmManf null with domain-deletion and CXXC point mutants plus mammalian neuron assays\",\n      \"pmids\": [\"24019940\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular partners of each domain not identified in this system\", \"Mechanistic basis of domain interdependence unresolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identified MANF as essential for pancreatic \\u03b2-cell proliferation and survival, linking its UPR-modulating function to a defined organ phenotype (diabetes).\",\n      \"evidence\": \"MANF knockout mice, recombinant MANF proliferation assays, and pancreatic overexpression in diabetic mice\",\n      \"pmids\": [\"24726366\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism of proliferative effect not defined\", \"Secreted vs. intracellular contributions not separated\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Extended MANF function beyond cytoprotection to neural progenitor migration/differentiation and central metabolic control, revealing context-dependent signaling outputs.\",\n      \"evidence\": \"MANF KO NSCs, in utero electroporation, STAT3/ERK Western blots, and hypothalamic viral gain/loss-of-function with PIP4k2b assays\",\n      \"pmids\": [\"29082311\", \"29050872\", \"28924165\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor mediating progenitor signaling not defined\", \"Connection between ER role and surface signaling unclear\", \"Generality across cell types untested\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Solved the central intracellular mechanism by showing the SAP domain binds ADP-bound BiP's nucleotide-binding domain and inhibits nucleotide exchange, stabilizing chaperone-client complexes.\",\n      \"evidence\": \"Crystal structures of MANF SAP\\u2013BiP NBD complex, in vitro nucleotide exchange assays, and BiP complex analysis in MANF-null cells\",\n      \"pmids\": [\"30710085\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular consequence of stabilized BiP complexes incompletely defined\", \"Whether BiP regulation accounts for all protective effects unresolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified Neuroplastin as a cell-surface MANF receptor coupling extracellular MANF to NF-\\u03baB suppression, providing a receptor for the secreted anti-inflammatory function.\",\n      \"evidence\": \"Co-immunoprecipitation, surface binding assays, NPTN manipulation, and NF-\\u03baB reporter assays\",\n      \"pmids\": [\"33299977\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab, no reciprocal in vivo receptor validation\", \"Downstream signaling between NPTN and NF-\\u03baB not mapped\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Established that MANF is a broad UPR regulator with an ER interactome beyond BiP, that ATP binding gates the BiP interaction, and that human LOF mutations cause a Mendelian disease, validating its physiological essentiality.\",\n      \"evidence\": \"AP-MS interactome, microscale thermophoresis/NMR ATP binding, neuronal mutant assays; human genetics with hESC \\u03b2-cell KO and xenotransplantation\",\n      \"pmids\": [\"33460650\", \"33500254\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional roles of most interactome partners undefined\", \"How distinct activities map to disease phenotypes unresolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined direct MANF\\u2013IRE1\\u03b1 binding as a BiP-independent mechanism that restrains the IRE1\\u03b1 UPR branch and is specifically required for dopaminergic neuroprotection in vivo.\",\n      \"evidence\": \"Co-IP, binding-interface mutagenesis, IRE1\\u03b1 oligomerization/phosphorylation and XBP1 splicing assays, and 6-OHDA PD model with MANF mutants\",\n      \"pmids\": [\"36739529\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative in vivo weighting of IRE1\\u03b1 vs. BiP mechanisms across tissues unresolved\", \"Structural basis of MANF-IRE1\\u03b1 interface not solved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed a non-canonical mitochondrial role in which de-SUMOylated MANF restores Parkin E3 ligase activity via its CXXC motif to drive mitophagy, expanding MANF function beyond the ER.\",\n      \"evidence\": \"SENP1 manipulation, MANF-PRKN Co-IP, ubiquitin ligase activity and mitophagy assays, CXXC mutagenesis in breast cancer cells\",\n      \"pmids\": [\"39147386\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab; physiological generality of mitochondrial MANF untested\", \"Trafficking route to mitochondria unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How MANF's distinct intracellular (BiP, IRE1\\u03b1, mitochondrial Parkin) and extracellular (Neuroplastin, S100A8) activities are coordinated and weighted within a given cell type to produce tissue-specific protection remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model integrating ER, secreted, and mitochondrial functions\", \"Receptor-to-signaling cascades incompletely mapped\", \"Determinants of tissue-specific phenotype selection unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [16, 27]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [16, 22]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [22, 16]},\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [20]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [1, 2, 16]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [2, 6]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6, 20]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [30]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [2, 16, 27]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [16, 22]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [10, 20, 29]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [28, 30]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [21, 14, 25]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"HSPA5\", \"ERN1\", \"NPTN\", \"S100A8\", \"XBP1\", \"PRKN\", \"MATN3\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}