{"gene":"GPSM1","run_date":"2026-06-10T01:55:21","timeline":{"discoveries":[{"year":2010,"finding":"AGS3 (GPSM1) forms a complex with Gαi1 in intact mammalian cells, as detected by BRET. Activation of α2-adrenergic receptors or μ-opioid receptors reduces this AGS3·Gαi1 complex by >30% in a pertussis-toxin- and RGS4-sensitive manner, but not by Gβγ sequestration. Upon receptor activation, AGS3 reversibly dissociates from the cortical signaling complex, demonstrating that a seven-transmembrane receptor directly regulates the AGS3·Gαi signaling module.","method":"Bioluminescence resonance energy transfer (BRET) in mammalian cells; pertussis toxin inhibition; RGS4 co-expression; GRK2-ct Gβγ sequestration","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — BRET-based complex formation in intact cells with multiple pharmacological and genetic controls (PTX, RGS4, GRK2-ct), single lab but orthogonal approaches","pmids":["20716524"],"is_preprint":false},{"year":2008,"finding":"AGS3 (GPSM1) is upregulated in the rat nucleus accumbens core during abstinence from ethanol self-administration. Knockdown of NAcore AGS3 reduces ethanol seeking to pre-abstinence levels. The effect is mediated through Gβγ: sequestration of Gβγ (but not Gαi knockdown) similarly reduces ethanol seeking, placing GPSM1 upstream of Gβγ-mediated signaling in compulsive ethanol seeking.","method":"Viral knockdown (siRNA) in vivo; operant ethanol self-administration behavioral model; Gβγ sequestration with GRK2-ct; Gαi knockdown","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo knockdown with behavioral readout combined with epistasis (Gβγ sequestration vs Gαi knockdown) to define pathway position","pmids":["18719114"],"is_preprint":false},{"year":2012,"finding":"GPSM1 increases heteromeric polycystin-1/polycystin-2 ion channel activity via Gβγ subunits in renal epithelial cells. Loss of GPSM1 in a Pkd1 hypomorphic mouse model accelerates cyst progression and reduces renal function, establishing GPSM1 as a modulator of polycystin channel activity and cyst progression via Gβγ.","method":"Electrophysiology (ion channel activity assay); genetic cross of Gpsm1 knockout with Pkd1^V/V mice; renal function assessment","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — direct electrophysiological assay of channel activity plus in vivo genetic model, single lab with two orthogonal methods","pmids":["23236168"],"is_preprint":false},{"year":2014,"finding":"AGS3 (GPSM1) is required for normal chemokine receptor signaling in leukocytes. AGS3-null B and T lymphocytes and bone marrow-derived dendritic cells exhibit significant chemotactic defects, reduced chemokine-stimulated calcium mobilization, and altered ERK and Akt activation, indicating AGS3 is a positive regulator of G-protein signaling downstream of chemokine receptors in immune cells.","method":"AGS3 knockout mice; chemotaxis assays; calcium mobilization assays; ERK and Akt western blotting","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean knockout with multiple orthogonal functional readouts (chemotaxis, Ca2+ flux, kinase activation), single lab","pmids":["24573680"],"is_preprint":false},{"year":2018,"finding":"Phosphorylation of a single threonine residue (T602) in the GPR domain of AGS3 (GPSM1) controls its subcellular distribution. Non-phosphorylatable AGS3-T602A localizes to cytosolic puncta rather than the cortical/diffuse distribution of wild-type AGS3. Co-expression of Gαi or Gαo (but not Gαs or Gαq) rescues the punctate localization. Alkaline phosphatase treatment confirms phosphorylation-dependent gel shifts, and the T602E/D phosphomimetics maintain punctate distribution, while T602A→T602 reversion restores normal localization.","method":"Site-directed mutagenesis; fluorescence microscopy; alkaline phosphatase treatment; SDS-PAGE gel shift; co-expression with Gα subunit variants","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — mutagenesis at defined residue with multiple point mutations and biochemical validation (gel shift), plus Gα rescue experiment; single lab with orthogonal approaches","pmids":["30404823"],"is_preprint":false},{"year":2022,"finding":"Myeloid GPSM1 promotes metabolic inflammation via the Gαi3/cAMP/PKA/CREB axis, which suppresses TNFAIP3 transcription and thereby sustains TLR4-induced NF-κB signaling in macrophages. Myeloid-specific GPSM1 ablation increases TNFAIP3 (A20) expression and inhibits NF-κB, protecting mice against high-fat-diet-induced insulin resistance, glucose dysregulation, and liver steatosis.","method":"Myeloid-specific conditional knockout mice; HFD metabolic phenotyping; cAMP/PKA/CREB pathway biochemistry; TNFAIP3 transcription assay; NF-κB signaling assays; small-molecule inhibitor (AN-465/42243987)","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — conditional KO in vivo with defined molecular pathway (Gαi3/cAMP/PKA/CREB/TNFAIP3/NF-κB) validated by multiple biochemical assays in single lab","pmids":["36434066"],"is_preprint":false},{"year":2020,"finding":"AGS3 (GPSM1) regulates trans-Golgi network (TGN) dynamics and the transport of TMED7-positive cargo containing E-cadherin (Cdh1) to the cell-contact surface during early mouse embryo development. CRISPR/Cas9-mediated AGS3 knockout causes developmental arrest, fragmentation after the four-cell stage, and decreased Cdh1 at cell-contact membranes, along with dispersal of TGN46- and TMED7-positive vesicles. Increased Gαi1 expression rescues AGS3-overexpression phenotypes, linking AGS3's GoLoco/Gαi interaction to TGN-dependent membrane trafficking.","method":"CRISPR/Cas9 knockout in mouse embryos; fluorescent protein tagging of TGN46 and TMED7; confocal live imaging; Gαi1 rescue overexpression","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with live imaging of cargo trafficking, plus Gαi1 rescue establishing functional link; single lab, multiple orthogonal methods","pmids":["33148610"],"is_preprint":false},{"year":2020,"finding":"AGS3 (GPSM1) inhibits Gαi3 activity, and this inhibition leads to markedly increased Bcl-2 phosphorylation on TGN38-containing intracellular vesicle pools. Loss of AGS3 (along with RGS4) results in reduced phospho-Bcl-2, whereas overexpression of AGS3 increases phospho-Bcl-2 levels, linking GPSM1 to autophagic signaling regulation via Gαi3-dependent Bcl-2 phosphorylation at TGN-associated membranes.","method":"Western blotting for phospho-Bcl-2; manipulation of palmitoylation/intracellular localization of RGS4; macrophage autophagy assays; adrenal gland autophagic flux in RGS4-KO mice","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — functional biochemical assays with localization data, single lab, indirect measure of GPSM1's role (via RGS4-KO background and AGS3 overexpression)","pmids":["32501280"],"is_preprint":false},{"year":2013,"finding":"AGS3 (GPSM1) enhances phosphorylation of CREB (p-CREB) in multiple myeloma cells. Knockdown of AGS3 reduces p-CREB levels, increases apoptosis, and reverses cell adhesion to fibronectin and HS-5 stromal cells, implicating GPSM1 in anti-apoptotic signaling via cAMP/PKA/CREB in a cell adhesion context.","method":"siRNA knockdown; western blotting for p-CREB; apoptosis assay; cell adhesion assay with fibronectin and HS-5 cells; doxorubicin-induced apoptosis model","journal":"International journal of hematology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — siRNA knockdown with multiple functional readouts (apoptosis, adhesion, p-CREB), single lab, consistent results across assays","pmids":["24307516"],"is_preprint":false},{"year":2015,"finding":"AGS3 (GPSM1) inhibits osteogenic differentiation of dental pulp stem cells exposed to TNF-α via the cAMP/PKA/CREB signaling pathway. Increased AGS3 expression suppresses p-CREB levels and downstream osteogenic gene expression (BMP2, ID2, osteocalcin), while knockdown of AGS3 promotes osteogenic differentiation.","method":"AGS3 knockdown (siRNA) and overexpression; osteogenic differentiation assays; western blotting for p-CREB and osteogenic markers; TNF-α stimulation","journal":"Differentiation; research in biological diversity","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — siRNA knockdown and overexpression with biochemical pathway readout, single lab, pathway confirmed by both gain- and loss-of-function","pmids":["26143356"],"is_preprint":false},{"year":2020,"finding":"Silencing of Gpsm1 in rat ovarian granulosa cells reduces cAMP levels, PKA catalytic subunit, and p-CREB, decreases the Bcl-2/Bax ratio, and increases Caspase-3/Cleaved Caspase-3, leading to increased apoptosis and decreased proliferation. This establishes GPSM1 as a pro-survival regulator in granulosa cells via the cAMP-PKA-CREB pathway.","method":"siRNA knockdown; CCK8 proliferation assay; flow cytometry (apoptosis); cAMP measurement; western blotting for PKAc, p-CREB, Bcl-2, Bax, Caspase-3","journal":"Journal of ovarian research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — siRNA KD with multiple biochemical readouts consistent with a defined pathway, single lab","pmids":["33220708"],"is_preprint":false},{"year":2023,"finding":"AGS3 (GPSM1) antagonizes LGN (GPSM2) to regulate spindle orientation and cell fate in mammalian epidermis. AGS3 overexpression displaces LGN from the apical cortex, increasing planar divisions and symmetric fates; AGS3 loss prolongs cortical LGN localization and biases divisions toward perpendicular orientation and asymmetric fates. Genetic epistasis in double mutants confirms AGS3 operates through LGN. Clonal lineage tracing shows that LGN promotes asymmetric fates while AGS3 promotes symmetric fates and influences differentiation via delamination.","method":"Static and ex vivo live imaging; conditional knockout and overexpression in mouse epidermis; genetic epistasis (double-mutant analysis); clonal lineage tracing","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo genetic epistasis with live imaging and clonal lineage tracing, multiple orthogonal methods, single lab","pmids":["37017303"],"is_preprint":false},{"year":2021,"finding":"Knockdown of GPSM1 in B-ALL cells inhibits proliferation and promotes apoptosis by suppressing the ADCY6-RAPGEF3-JNK signaling pathway. Reduced GPSM1 decreases expression of ADCY6 and RAPGEF3, with downstream reduction in JNK activity, linking GPSM1 to cAMP production and JNK-mediated survival signaling.","method":"siRNA knockdown; cell proliferation and apoptosis assays; western blotting for ADCY6, RAPGEF3, JNK; cell cycle analysis","journal":"Pathology oncology research : POR","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — siRNA knockdown with multiple downstream pathway readouts, single lab, mechanistic connection to ADCY6-RAPGEF3-JNK pathway","pmids":["34257610"],"is_preprint":false},{"year":2023,"finding":"GPSM1 deficiency in POMC neurons enhances autophagy and leptin sensitivity through the PI3K/AKT/mTOR signaling pathway, increasing POMC expression and α-MSH production, enhancing sympathetic innervation, and increasing brown adipose tissue thermogenesis. POMC-specific GPSM1 knockout mice are protected against diet-induced obesity, glucose dysregulation, and insulin resistance.","method":"POMC neuron-specific conditional knockout mice; HFD metabolic phenotyping; molecular/biochemical analysis of PI3K/AKT/mTOR pathway; autophagy assays; immunofluorescence/immunohistochemistry; sympathetic innervation analysis","journal":"Molecular metabolism","confidence":"High","confidence_rationale":"Tier 2 / Moderate — cell-type-specific conditional KO with defined molecular pathway (PI3K/AKT/mTOR/autophagy/leptin sensitivity) and multiple in vivo and in vitro readouts, single lab","pmids":["37979657"],"is_preprint":false},{"year":2023,"finding":"GPSM1 promotes colorectal cancer cell migration and invasion and inhibits autophagy by activating the PI3K/AKT/mTOR pathway. GFP-LC3B imaging and autophagic vesicle ultrastructure confirmed that GPSM1 suppresses autophagic flux, while gain- and loss-of-function experiments in vitro and a tumor metastasis mouse model confirmed its role in metastasis.","method":"siRNA knockdown and overexpression; GFP-LC3B immunofluorescence; electron microscopy of autophagic vesicles; PI3K/AKT/mTOR western blotting; in vivo metastasis mouse model","journal":"The international journal of biochemistry & cell biology","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — gain- and loss-of-function with multiple orthogonal autophagy detection methods, single lab","pmids":["36758790"],"is_preprint":false},{"year":2025,"finding":"GPSM1 in macrophages promotes anti-PD-1 resistance in colorectal cancer by driving M2 polarization and tumor microenvironment immunosuppression. The deubiquitinase USP9X stabilizes GPSM1 by preventing K63-polyubiquitination-mediated degradation. Stabilized GPSM1 promotes MEIS3 nuclear translocation, which activates macrophage colony-stimulating factor (CSF1) expression.","method":"ChIP-PCR; mass spectrometry; co-immunoprecipitation; single-cell RNA sequencing; multiplex immunofluorescence; orthotopic CRC xenograft model; mass cytometry; flow cytometry","journal":"Journal for immunotherapy of cancer","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — ChIP-PCR, mass spectrometry, and co-immunoprecipitation to establish USP9X-GPSM1-MEIS3-CSF1 axis, with in vivo validation; single lab but multiple orthogonal methods","pmids":["40010765"],"is_preprint":false},{"year":2025,"finding":"Myeloid GPSM1 promotes atherosclerosis by sustaining monocyte activation, chemotaxis, and adhesion through the cAMP/PKA/KLF4/PMP22 axis, which activates the p38/ERK MAPK pathway. Myeloid-specific GPSM1 ablation protects against atherosclerosis in Apoe-/- and AAV-PCSK9 mice, while myeloid-restricted GPSM1 overexpression accelerates disease. siRNA-loaded liposome blockade of PMP22 rescues GPSM1-overexpression mice, and a small molecule inhibiting GPSM1 suppresses atherosclerosis in vivo.","method":"Myeloid-specific conditional knockout and overexpression in Apoe-/- and AAV-PCSK9 mouse models; monocyte chemotaxis and adhesion assays; cAMP/PKA/KLF4/PMP22 pathway biochemistry; p38/ERK MAPK western blotting; siRNA-loaded liposome PMP22 blockade; small-molecule GPSM1 inhibitor","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — conditional KO and OE in vivo with defined molecular pathway plus rescue experiment (PMP22 blockade) and pharmacological validation, single lab with multiple orthogonal approaches","pmids":["41296728"],"is_preprint":false},{"year":2024,"finding":"AGS3 (GPSM1) forms biomolecular condensates (BMCs) under cellular stress (oxidative, pH, thermal). Stress-induced AGS3 BMCs are biochemically distinct from G3BP1 stress granules and Dcp1a P-bodies. Co-expression of Gαi3 (but not DVL2) reduces stress-induced BMC formation. FRAP analysis reveals distinct diffusion kinetics and restricted fluidity within AGS3 BMCs, and stress shifts AGS3 to a membrane pellet fraction.","method":"Fluorescence microscopy; FRAP; immunoblotting of fractionated cell lysates; co-expression experiments with Gαi3 and DVL2; stress induction (oxidative, pH, thermal)","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — FRAP and fractionation with multiple stress conditions and Gαi3 rescue, single lab","pmids":["38264908"],"is_preprint":false},{"year":2020,"finding":"AGS3 (GPSM1) interacts with Dishevelled-2 (DVL2), and this interaction is regulated by protein phosphorylation, subcellular distribution, and a cell-surface GPCR. AGS3 signaling influences β-catenin-regulated transcription through the WNT-Frizzled-Dishevelled axis, suggesting integration between Gαi/GPR signaling and WNT pathway.","method":"Co-immunoprecipitation; phosphorylation manipulation; GPCR activation; β-catenin transcription reporter assay; subcellular localization imaging","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP interaction and reporter assay for β-catenin transcription, single lab, single paper","pmids":["32737219"],"is_preprint":false},{"year":2013,"finding":"Macrophages lacking Gαi3, AGS3 (GPSM1), or RGS19, or treated with pertussis toxin, show normal levels of basal autophagy, autophagic induction, autophagic flux, autophagic degradation, and anti-autophagic action, indicating that the Gαi3/AGS3/RGS19 pathway does not regulate autophagy in macrophages (negative finding).","method":"LC3 processing western blot; LC3 puncta formation assay; long-lived protein degradation assay; Gpsm1-/-, Gnai3-/-, Rgs19-/- bone marrow-derived macrophages; pertussis toxin treatment","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal autophagy assays in knockout macrophages; negative result replicated across multiple genotypes; single lab","pmids":["24312373"],"is_preprint":false},{"year":2026,"finding":"GPSM1 in CD4+ T cells (especially Treg cells) restricts CD73+CD103+ Treg cell abundance in adipose tissue. GPSM1 deletion in CD4+ T cells or Treg cells increases adipose Treg cells, reduces inflammation, and improves metabolic parameters. Mechanistically, GPSM1 regulates Treg cell abundance via a RHOA-cell stiffness-TAZ axis. Adoptive transfer of GPSM1-deficient Treg cells improves energy expenditure and glucose/lipid metabolism in Rag1-/- mice.","method":"Conditional knockout (CD4-Cre and Treg-specific); CD4-T-cell-specific overexpression; flow cytometry (Treg subset analysis); HFD metabolic phenotyping; adoptive transfer into Rag1-/- mice; RHOA/TAZ pathway biochemistry","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — cell-type-specific KO and OE with adoptive transfer rescue and RHOA-TAZ mechanistic pathway, multiple orthogonal methods, single lab","pmids":["42185270"],"is_preprint":false}],"current_model":"GPSM1 (AGS3) is a receptor-independent G-protein signaling modulator whose GoLoco/GPR motifs bind Gαi-GDP free of Gβγ, allowing it to both inhibit Gαi-mediated signaling and release Gβγ for downstream signaling; its subcellular distribution is dynamically regulated by phosphorylation of T602 and interaction with Gαi/o subunits, and it functions in diverse cellular contexts including immune cell chemotaxis (via Gβγ), TGN-to-membrane cargo trafficking of E-cadherin, epidermal spindle orientation (by antagonizing LGN/GPSM2 at the apical cortex), hypothalamic POMC neuron energy balance (via PI3K/AKT/mTOR and autophagy), myeloid cell pro-inflammatory signaling (via Gαi3/cAMP/PKA/CREB/TNFAIP3/NF-κB and cAMP/PKA/KLF4/PMP22/MAPK axes), T-cell/Treg homeostasis (via RHOA-cell stiffness-TAZ), and tumor microenvironment immunosuppression (via USP9X-stabilized GPSM1 driving MEIS3/CSF1 expression)."},"narrative":{"mechanistic_narrative":"GPSM1 (AGS3) is a receptor-independent activator of G-protein signaling that uses its GoLoco/GPR motifs to bind Gαi/o-GDP, thereby partitioning heterotrimeric G-protein subunits and tuning both Gαi-dependent and Gβγ-dependent outputs across many cell types [PMID:20716524, PMID:30404823]. In intact cells GPSM1 assembles with Gαi1 and dynamically dissociates from a cortical signaling complex upon GPCR activation in a pertussis-toxin- and RGS4-sensitive manner [PMID:20716524], and its subcellular distribution is governed by phosphorylation of a single GPR-domain threonine (T602), with Gαi or Gαo co-expression—but not Gαs or Gαq—redirecting its localization [PMID:30404823]. Through the released Gβγ arm, GPSM1 acts as a positive regulator of chemokine-receptor signaling, supporting leukocyte chemotaxis, calcium mobilization, and ERK/Akt activation [PMID:24573680], potentiates polycystin-1/polycystin-2 channel activity to restrain renal cyst progression [PMID:23236168], and drives compulsive ethanol-seeking behavior [PMID:18719114]. A recurrent mechanistic theme is GPSM1 control of the cAMP/PKA/CREB axis: in myeloid cells GPSM1 sustains TLR4-induced NF-κB inflammation via Gαi3/cAMP/PKA/CREB-mediated suppression of TNFAIP3, promoting diet-induced metabolic disease [PMID:36434066], and drives monocyte activation and atherosclerosis through a cAMP/PKA/KLF4/PMP22–p38/ERK MAPK cascade [PMID:41296728]. GPSM1 also localizes to and organizes trans-Golgi network membranes, where it directs TMED7/E-cadherin cargo trafficking required for early embryonic cell contacts [PMID:33148610], and it antagonizes its paralog LGN/GPSM2 at the apical cortex to bias epidermal spindle orientation and cell fate [PMID:37017303]. In immunometabolic and oncogenic settings, GPSM1 modulates PI3K/AKT/mTOR signaling and autophagy in POMC neurons and colorectal cancer cells [PMID:37979657, PMID:36758790], shapes Treg homeostasis via a RHOA–cell-stiffness–TAZ axis [PMID:42185270], and—when stabilized by the deubiquitinase USP9X—promotes MEIS3-driven CSF1 expression and tumor immunosuppression [PMID:40010765].","teleology":[{"year":2008,"claim":"Established that GPSM1 acts upstream of Gβγ signaling in a behavioral context, defining its pathway position rather than merely correlating its expression with a phenotype.","evidence":"Viral siRNA knockdown in rat nucleus accumbens with operant ethanol self-administration and Gβγ-sequestration epistasis","pmids":["18719114"],"confidence":"High","gaps":["Does not resolve which GPCR engages the GPSM1·Gαi module in this circuit","Molecular partners of the released Gβγ in NAcore neurons not identified"]},{"year":2010,"claim":"Showed that a seven-transmembrane receptor directly regulates the GPSM1·Gαi module, answering whether this 'receptor-independent' modulator is itself responsive to GPCR activation in living cells.","evidence":"BRET in mammalian cells with α2-adrenergic/μ-opioid receptor activation, pertussis toxin, RGS4, and GRK2-ct controls","pmids":["20716524"],"confidence":"High","gaps":["Mechanism by which receptor activation triggers cortical complex dissociation not defined","Stoichiometry of the GPSM1·Gαi·receptor assembly unknown"]},{"year":2012,"claim":"Demonstrated a physiological output for GPSM1-released Gβγ on an ion channel, linking the modulator to organ-level disease progression.","evidence":"Electrophysiology of polycystin-1/2 channels plus Gpsm1-knockout × Pkd1 hypomorph mouse cross","pmids":["23236168"],"confidence":"High","gaps":["Direct Gβγ–polycystin interaction not structurally defined","Whether GPSM1 acts at the channel locale or upstream is unresolved"]},{"year":2013,"claim":"Defined GPSM1 as a positive regulator of immune-cell chemokine receptor signaling, clarifying the direction of its effect on G-protein output.","evidence":"AGS3-knockout B/T lymphocytes and dendritic cells with chemotaxis, calcium flux, and ERK/Akt assays","pmids":["24573680"],"confidence":"High","gaps":["Which chemokine receptors physically couple to GPSM1 not mapped","Relative contribution of Gαi vs Gβγ arms not separated in these cells"]},{"year":2013,"claim":"Connected GPSM1 to cAMP/PKA/CREB anti-apoptotic and adhesion signaling in a cancer context, beginning a recurring pathway theme.","evidence":"siRNA knockdown in multiple myeloma cells with p-CREB, apoptosis, and fibronectin/HS-5 adhesion assays","pmids":["24307516"],"confidence":"Medium","gaps":["Direct link from GPSM1/Gαi to adenylyl cyclase not established","Single cell-line system without in vivo validation"]},{"year":2013,"claim":"Provided a negative control on the autophagy hypothesis, showing the Gαi3/AGS3/RGS19 module is dispensable for macrophage autophagy under the conditions tested.","evidence":"LC3 processing, puncta, and long-lived protein degradation assays in Gpsm1-/-, Gnai3-/-, Rgs19-/- macrophages with pertussis toxin","pmids":["24312373"],"confidence":"Medium","gaps":["Negative result is context-specific to macrophages and may not generalize to other cell types","Does not exclude autophagy roles in non-myeloid cells"]},{"year":2015,"claim":"Extended the cAMP/PKA/CREB axis to differentiation control, showing GPSM1 suppresses osteogenic gene programs.","evidence":"siRNA knockdown and overexpression in TNF-α-treated dental pulp stem cells with p-CREB and osteogenic marker readouts","pmids":["26143356"],"confidence":"Medium","gaps":["G-protein step linking GPSM1 to CREB not biochemically dissected","In vivo relevance not tested"]},{"year":2018,"claim":"Identified a single phosphorylation switch (T602) and Gαi/o binding as the determinants of GPSM1 subcellular distribution, providing a mechanistic basis for its dynamic localization.","evidence":"Site-directed mutagenesis (T602A/E/D and reversion), fluorescence microscopy, alkaline phosphatase gel shift, and Gα-subunit co-expression rescue","pmids":["30404823"],"confidence":"High","gaps":["Kinase responsible for T602 phosphorylation not identified","Functional consequence of punctate vs cortical distribution for signaling not directly measured"]},{"year":2020,"claim":"Placed GPSM1 at the trans-Golgi network controlling E-cadherin cargo trafficking, expanding its role beyond plasma-membrane G-protein signaling to membrane logistics.","evidence":"CRISPR/Cas9 knockout in mouse embryos with TGN46/TMED7 live imaging and Gαi1 rescue","pmids":["33148610"],"confidence":"High","gaps":["Molecular machinery linking GPSM1·Gαi to TMED7 cargo selection not defined","Whether Gβγ release participates in TGN trafficking unresolved"]},{"year":2020,"claim":"Linked GPSM1's inhibition of Gαi3 to Bcl-2 phosphorylation at TGN-associated membranes, suggesting a route to autophagic signaling regulation.","evidence":"Phospho-Bcl-2 immunoblotting with AGS3 overexpression/loss in an RGS4-KO background and adrenal autophagic flux","pmids":["32501280"],"confidence":"Medium","gaps":["GPSM1 effect inferred indirectly through RGS4 manipulation","Causal chain from phospho-Bcl-2 to autophagy outcome not established"]},{"year":2020,"claim":"Connected GPSM1 to WNT/β-catenin signaling through a phosphorylation- and GPCR-regulated interaction with DVL2.","evidence":"Co-immunoprecipitation, phosphorylation manipulation, and β-catenin transcription reporter assay","pmids":["32737219"],"confidence":"Medium","gaps":["Single Co-IP without reciprocal structural validation","Direct effect of GPSM1 on Frizzled–Dishevelled assembly not shown"]},{"year":2020,"claim":"Established GPSM1 as a pro-survival regulator in granulosa cells operating through cAMP-PKA-CREB and Bcl-2/Bax balance.","evidence":"siRNA knockdown in rat granulosa cells with cAMP measurement, proliferation, apoptosis, and pathway immunoblots","pmids":["33220708"],"confidence":"Medium","gaps":["G-protein step coupling GPSM1 to cAMP not dissected","Single-species, single-cell-type model"]},{"year":2021,"claim":"Tied GPSM1 to a cAMP-generating ADCY6-RAPGEF3-JNK survival pathway in leukemia cells.","evidence":"siRNA knockdown in B-ALL cells with proliferation, apoptosis, cell-cycle, and ADCY6/RAPGEF3/JNK immunoblots","pmids":["34257610"],"confidence":"Medium","gaps":["Mechanism by which GPSM1 controls ADCY6 expression unknown","No in vivo confirmation"]},{"year":2022,"claim":"Defined a complete myeloid signaling cascade by which GPSM1 sustains inflammation, answering how it drives metabolic disease at the molecular level.","evidence":"Myeloid-specific conditional knockout mice on HFD with Gαi3/cAMP/PKA/CREB/TNFAIP3/NF-κB biochemistry and small-molecule inhibitor","pmids":["36434066"],"confidence":"High","gaps":["Upstream receptor coupling GPSM1 to Gαi3 in macrophages not identified","Transcriptional mechanism of CREB-mediated TNFAIP3 repression not fully mapped"]},{"year":2023,"claim":"Showed GPSM1 antagonizes its paralog LGN/GPSM2 at the apical cortex to set spindle orientation and cell fate, defining a paralog-competition mechanism.","evidence":"Conditional knockout/overexpression in mouse epidermis with live imaging, double-mutant epistasis, and clonal lineage tracing","pmids":["37017303"],"confidence":"High","gaps":["Molecular basis of cortical LGN displacement by GPSM1 not resolved","Role of Gαi in this cortical competition not directly tested"]},{"year":2023,"claim":"Established a neuronal role for GPSM1 in energy balance via PI3K/AKT/mTOR-dependent autophagy and leptin sensitivity in POMC neurons.","evidence":"POMC-specific conditional knockout mice with HFD phenotyping, autophagy assays, and sympathetic innervation/BAT thermogenesis analysis","pmids":["37979657"],"confidence":"High","gaps":["How GPSM1 G-protein activity feeds into PI3K/AKT/mTOR not defined","Direct substrate or effector connecting GPSM1 to autophagy machinery unknown"]},{"year":2023,"claim":"Showed GPSM1 suppresses autophagic flux and promotes metastasis in colorectal cancer through PI3K/AKT/mTOR activation.","evidence":"Gain/loss-of-function with GFP-LC3B imaging, autophagic vesicle electron microscopy, pathway immunoblots, and a metastasis mouse model","pmids":["36758790"],"confidence":"Medium","gaps":["Mechanistic link from GPSM1 to PI3K activation not defined","Whether autophagy suppression is causal for metastasis not separated"]},{"year":2024,"claim":"Revealed that GPSM1 forms stress-induced biomolecular condensates distinct from stress granules and P-bodies, adding a phase-separation behavior regulated by Gαi3.","evidence":"Fluorescence microscopy, FRAP, lysate fractionation, and G αi3/DVL2 co-expression under oxidative/pH/thermal stress","pmids":["38264908"],"confidence":"Medium","gaps":["Functional role of GPSM1 condensates in signaling or stress response unknown","Composition of the condensates not characterized"]},{"year":2025,"claim":"Defined a deubiquitination-driven oncogenic axis in which USP9X stabilizes GPSM1 to promote MEIS3/CSF1-mediated tumor immunosuppression and immunotherapy resistance.","evidence":"Mass spectrometry, co-immunoprecipitation, ChIP-PCR, single-cell RNA-seq, and orthotopic CRC xenografts with mass/flow cytometry","pmids":["40010765"],"confidence":"High","gaps":["How GPSM1 promotes MEIS3 nuclear translocation mechanistically not resolved","Whether G-protein modulator activity is required for the MEIS3/CSF1 output unclear"]},{"year":2025,"claim":"Identified a complete myeloid cAMP/PKA/KLF4/PMP22–MAPK cascade through which GPSM1 drives atherosclerosis, and validated it pharmacologically and by rescue.","evidence":"Myeloid conditional KO and overexpression in Apoe-/- and AAV-PCSK9 mice with chemotaxis/adhesion assays, pathway biochemistry, PMP22 siRNA-liposome rescue, and a small-molecule GPSM1 inhibitor","pmids":["41296728"],"confidence":"High","gaps":["Upstream receptor/Gα coupling in monocytes not defined","Relationship between this KLF4/PMP22 cascade and the TNFAIP3/NF-κB cascade in myeloid cells not reconciled"]},{"year":2026,"claim":"Established GPSM1 as a restrictor of adipose Treg homeostasis acting through a RHOA–cell-stiffness–TAZ mechanotransduction axis, with metabolic consequences confirmed by adoptive transfer.","evidence":"CD4/Treg-specific conditional KO and overexpression with HFD phenotyping, Treg subset flow cytometry, RHOA/TAZ biochemistry, and adoptive transfer into Rag1-/- mice","pmids":["42185270"],"confidence":"High","gaps":["How GPSM1 G-protein activity engages RHOA/cell stiffness not defined","Direct GPSM1 effectors in Treg cells not identified"]},{"year":null,"claim":"The unifying question remains how a single GoLoco/GPR Gαi/o modulator selects among its many divergent outputs—Gβγ-driven chemotaxis and channel modulation, cAMP/PKA/CREB inflammation, PI3K/AKT/mTOR autophagy, TGN cargo trafficking, cortical LGN antagonism, and ubiquitin-stabilized transcriptional programs—and which upstream receptors and cell-type-specific cofactors dictate this choice.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model reconciling Gαi-binding with the diverse downstream cascades","Upstream GPCRs coupling GPSM1 in most contexts unidentified","Determinants of context-specific pathway selection unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,3,4,5]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,2,3]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,4,6]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[4,17]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,6,11]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[6,7]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,3,4,5]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[3,5,15,20]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[13,14]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[6]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[6,11]}],"complexes":[],"partners":["GNAI1","GNAI3","GNAO1","DVL2","GPSM2","USP9X","RGS4"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86YR5","full_name":"G-protein-signaling modulator 1","aliases":["Activator of G-protein signaling 3"],"length_aa":675,"mass_kda":74.5,"function":"Guanine nucleotide dissociation inhibitor (GDI) which functions as a receptor-independent activator of heterotrimeric G-protein signaling. Keeps G(i/o) alpha subunit in its GDP-bound form thus uncoupling heterotrimeric G-proteins signaling from G protein-coupled receptors. Controls spindle orientation and asymmetric cell fate of cerebral cortical progenitors. May also be involved in macroautophagy in intestinal cells. May play a role in drug addiction","subcellular_location":"Cytoplasm, cytosol; Endoplasmic reticulum membrane; Golgi apparatus membrane; Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q86YR5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/GPSM1","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/GPSM1","total_profiled":1310},"omim":[{"mim_id":"616919","title":"FERM AND PDZ DOMAINS-CONTAINING PROTEIN 1; FRMPD1","url":"https://www.omim.org/entry/616919"},{"mim_id":"610668","title":"INSC SPINDLE ORIENTATION ADAPTOR PROTEIN; INSC","url":"https://www.omim.org/entry/610668"},{"mim_id":"609491","title":"G PROTEIN SIGNALING MODULATOR 1; GPSM1","url":"https://www.omim.org/entry/609491"},{"mim_id":"609245","title":"G PROTEIN SIGNALING MODULATOR 2; GPSM2","url":"https://www.omim.org/entry/609245"},{"mim_id":"266600","title":"INFLAMMATORY BOWEL DISEASE (CROHN DISEASE) 1; IBD1","url":"https://www.omim.org/entry/266600"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Golgi apparatus","reliability":"Supported"},{"location":"Nucleoplasm","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"brain","ntpm":105.6}],"url":"https://www.proteinatlas.org/search/GPSM1"},"hgnc":{"alias_symbol":["AGS3","DKFZP727I051"],"prev_symbol":[]},"alphafold":{"accession":"Q86YR5","domains":[{"cath_id":"1.25.40.10","chopping":"285-389_632-638","consensus_level":"medium","plddt":86.3621,"start":285,"end":638}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86YR5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86YR5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86YR5-F1-predicted_aligned_error_v6.png","plddt_mean":67.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=GPSM1","jax_strain_url":"https://www.jax.org/strain/search?query=GPSM1"},"sequence":{"accession":"Q86YR5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86YR5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86YR5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86YR5"}},"corpus_meta":[{"pmid":"29947923","id":"PMC_29947923","title":"Genetic 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Activation of α2-adrenergic receptors or μ-opioid receptors reduces this AGS3·Gαi1 complex by >30% in a pertussis-toxin- and RGS4-sensitive manner, but not by Gβγ sequestration. Upon receptor activation, AGS3 reversibly dissociates from the cortical signaling complex, demonstrating that a seven-transmembrane receptor directly regulates the AGS3·Gαi signaling module.\",\n      \"method\": \"Bioluminescence resonance energy transfer (BRET) in mammalian cells; pertussis toxin inhibition; RGS4 co-expression; GRK2-ct Gβγ sequestration\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — BRET-based complex formation in intact cells with multiple pharmacological and genetic controls (PTX, RGS4, GRK2-ct), single lab but orthogonal approaches\",\n      \"pmids\": [\"20716524\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"AGS3 (GPSM1) is upregulated in the rat nucleus accumbens core during abstinence from ethanol self-administration. Knockdown of NAcore AGS3 reduces ethanol seeking to pre-abstinence levels. The effect is mediated through Gβγ: sequestration of Gβγ (but not Gαi knockdown) similarly reduces ethanol seeking, placing GPSM1 upstream of Gβγ-mediated signaling in compulsive ethanol seeking.\",\n      \"method\": \"Viral knockdown (siRNA) in vivo; operant ethanol self-administration behavioral model; Gβγ sequestration with GRK2-ct; Gαi knockdown\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo knockdown with behavioral readout combined with epistasis (Gβγ sequestration vs Gαi knockdown) to define pathway position\",\n      \"pmids\": [\"18719114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"GPSM1 increases heteromeric polycystin-1/polycystin-2 ion channel activity via Gβγ subunits in renal epithelial cells. Loss of GPSM1 in a Pkd1 hypomorphic mouse model accelerates cyst progression and reduces renal function, establishing GPSM1 as a modulator of polycystin channel activity and cyst progression via Gβγ.\",\n      \"method\": \"Electrophysiology (ion channel activity assay); genetic cross of Gpsm1 knockout with Pkd1^V/V mice; renal function assessment\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — direct electrophysiological assay of channel activity plus in vivo genetic model, single lab with two orthogonal methods\",\n      \"pmids\": [\"23236168\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"AGS3 (GPSM1) is required for normal chemokine receptor signaling in leukocytes. AGS3-null B and T lymphocytes and bone marrow-derived dendritic cells exhibit significant chemotactic defects, reduced chemokine-stimulated calcium mobilization, and altered ERK and Akt activation, indicating AGS3 is a positive regulator of G-protein signaling downstream of chemokine receptors in immune cells.\",\n      \"method\": \"AGS3 knockout mice; chemotaxis assays; calcium mobilization assays; ERK and Akt western blotting\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean knockout with multiple orthogonal functional readouts (chemotaxis, Ca2+ flux, kinase activation), single lab\",\n      \"pmids\": [\"24573680\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Phosphorylation of a single threonine residue (T602) in the GPR domain of AGS3 (GPSM1) controls its subcellular distribution. Non-phosphorylatable AGS3-T602A localizes to cytosolic puncta rather than the cortical/diffuse distribution of wild-type AGS3. Co-expression of Gαi or Gαo (but not Gαs or Gαq) rescues the punctate localization. Alkaline phosphatase treatment confirms phosphorylation-dependent gel shifts, and the T602E/D phosphomimetics maintain punctate distribution, while T602A→T602 reversion restores normal localization.\",\n      \"method\": \"Site-directed mutagenesis; fluorescence microscopy; alkaline phosphatase treatment; SDS-PAGE gel shift; co-expression with Gα subunit variants\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — mutagenesis at defined residue with multiple point mutations and biochemical validation (gel shift), plus Gα rescue experiment; single lab with orthogonal approaches\",\n      \"pmids\": [\"30404823\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Myeloid GPSM1 promotes metabolic inflammation via the Gαi3/cAMP/PKA/CREB axis, which suppresses TNFAIP3 transcription and thereby sustains TLR4-induced NF-κB signaling in macrophages. Myeloid-specific GPSM1 ablation increases TNFAIP3 (A20) expression and inhibits NF-κB, protecting mice against high-fat-diet-induced insulin resistance, glucose dysregulation, and liver steatosis.\",\n      \"method\": \"Myeloid-specific conditional knockout mice; HFD metabolic phenotyping; cAMP/PKA/CREB pathway biochemistry; TNFAIP3 transcription assay; NF-κB signaling assays; small-molecule inhibitor (AN-465/42243987)\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO in vivo with defined molecular pathway (Gαi3/cAMP/PKA/CREB/TNFAIP3/NF-κB) validated by multiple biochemical assays in single lab\",\n      \"pmids\": [\"36434066\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AGS3 (GPSM1) regulates trans-Golgi network (TGN) dynamics and the transport of TMED7-positive cargo containing E-cadherin (Cdh1) to the cell-contact surface during early mouse embryo development. CRISPR/Cas9-mediated AGS3 knockout causes developmental arrest, fragmentation after the four-cell stage, and decreased Cdh1 at cell-contact membranes, along with dispersal of TGN46- and TMED7-positive vesicles. Increased Gαi1 expression rescues AGS3-overexpression phenotypes, linking AGS3's GoLoco/Gαi interaction to TGN-dependent membrane trafficking.\",\n      \"method\": \"CRISPR/Cas9 knockout in mouse embryos; fluorescent protein tagging of TGN46 and TMED7; confocal live imaging; Gαi1 rescue overexpression\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with live imaging of cargo trafficking, plus Gαi1 rescue establishing functional link; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"33148610\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AGS3 (GPSM1) inhibits Gαi3 activity, and this inhibition leads to markedly increased Bcl-2 phosphorylation on TGN38-containing intracellular vesicle pools. Loss of AGS3 (along with RGS4) results in reduced phospho-Bcl-2, whereas overexpression of AGS3 increases phospho-Bcl-2 levels, linking GPSM1 to autophagic signaling regulation via Gαi3-dependent Bcl-2 phosphorylation at TGN-associated membranes.\",\n      \"method\": \"Western blotting for phospho-Bcl-2; manipulation of palmitoylation/intracellular localization of RGS4; macrophage autophagy assays; adrenal gland autophagic flux in RGS4-KO mice\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — functional biochemical assays with localization data, single lab, indirect measure of GPSM1's role (via RGS4-KO background and AGS3 overexpression)\",\n      \"pmids\": [\"32501280\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"AGS3 (GPSM1) enhances phosphorylation of CREB (p-CREB) in multiple myeloma cells. Knockdown of AGS3 reduces p-CREB levels, increases apoptosis, and reverses cell adhesion to fibronectin and HS-5 stromal cells, implicating GPSM1 in anti-apoptotic signaling via cAMP/PKA/CREB in a cell adhesion context.\",\n      \"method\": \"siRNA knockdown; western blotting for p-CREB; apoptosis assay; cell adhesion assay with fibronectin and HS-5 cells; doxorubicin-induced apoptosis model\",\n      \"journal\": \"International journal of hematology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — siRNA knockdown with multiple functional readouts (apoptosis, adhesion, p-CREB), single lab, consistent results across assays\",\n      \"pmids\": [\"24307516\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"AGS3 (GPSM1) inhibits osteogenic differentiation of dental pulp stem cells exposed to TNF-α via the cAMP/PKA/CREB signaling pathway. Increased AGS3 expression suppresses p-CREB levels and downstream osteogenic gene expression (BMP2, ID2, osteocalcin), while knockdown of AGS3 promotes osteogenic differentiation.\",\n      \"method\": \"AGS3 knockdown (siRNA) and overexpression; osteogenic differentiation assays; western blotting for p-CREB and osteogenic markers; TNF-α stimulation\",\n      \"journal\": \"Differentiation; research in biological diversity\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — siRNA knockdown and overexpression with biochemical pathway readout, single lab, pathway confirmed by both gain- and loss-of-function\",\n      \"pmids\": [\"26143356\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Silencing of Gpsm1 in rat ovarian granulosa cells reduces cAMP levels, PKA catalytic subunit, and p-CREB, decreases the Bcl-2/Bax ratio, and increases Caspase-3/Cleaved Caspase-3, leading to increased apoptosis and decreased proliferation. This establishes GPSM1 as a pro-survival regulator in granulosa cells via the cAMP-PKA-CREB pathway.\",\n      \"method\": \"siRNA knockdown; CCK8 proliferation assay; flow cytometry (apoptosis); cAMP measurement; western blotting for PKAc, p-CREB, Bcl-2, Bax, Caspase-3\",\n      \"journal\": \"Journal of ovarian research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — siRNA KD with multiple biochemical readouts consistent with a defined pathway, single lab\",\n      \"pmids\": [\"33220708\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"AGS3 (GPSM1) antagonizes LGN (GPSM2) to regulate spindle orientation and cell fate in mammalian epidermis. AGS3 overexpression displaces LGN from the apical cortex, increasing planar divisions and symmetric fates; AGS3 loss prolongs cortical LGN localization and biases divisions toward perpendicular orientation and asymmetric fates. Genetic epistasis in double mutants confirms AGS3 operates through LGN. Clonal lineage tracing shows that LGN promotes asymmetric fates while AGS3 promotes symmetric fates and influences differentiation via delamination.\",\n      \"method\": \"Static and ex vivo live imaging; conditional knockout and overexpression in mouse epidermis; genetic epistasis (double-mutant analysis); clonal lineage tracing\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic epistasis with live imaging and clonal lineage tracing, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"37017303\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Knockdown of GPSM1 in B-ALL cells inhibits proliferation and promotes apoptosis by suppressing the ADCY6-RAPGEF3-JNK signaling pathway. Reduced GPSM1 decreases expression of ADCY6 and RAPGEF3, with downstream reduction in JNK activity, linking GPSM1 to cAMP production and JNK-mediated survival signaling.\",\n      \"method\": \"siRNA knockdown; cell proliferation and apoptosis assays; western blotting for ADCY6, RAPGEF3, JNK; cell cycle analysis\",\n      \"journal\": \"Pathology oncology research : POR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — siRNA knockdown with multiple downstream pathway readouts, single lab, mechanistic connection to ADCY6-RAPGEF3-JNK pathway\",\n      \"pmids\": [\"34257610\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"GPSM1 deficiency in POMC neurons enhances autophagy and leptin sensitivity through the PI3K/AKT/mTOR signaling pathway, increasing POMC expression and α-MSH production, enhancing sympathetic innervation, and increasing brown adipose tissue thermogenesis. POMC-specific GPSM1 knockout mice are protected against diet-induced obesity, glucose dysregulation, and insulin resistance.\",\n      \"method\": \"POMC neuron-specific conditional knockout mice; HFD metabolic phenotyping; molecular/biochemical analysis of PI3K/AKT/mTOR pathway; autophagy assays; immunofluorescence/immunohistochemistry; sympathetic innervation analysis\",\n      \"journal\": \"Molecular metabolism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-type-specific conditional KO with defined molecular pathway (PI3K/AKT/mTOR/autophagy/leptin sensitivity) and multiple in vivo and in vitro readouts, single lab\",\n      \"pmids\": [\"37979657\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"GPSM1 promotes colorectal cancer cell migration and invasion and inhibits autophagy by activating the PI3K/AKT/mTOR pathway. GFP-LC3B imaging and autophagic vesicle ultrastructure confirmed that GPSM1 suppresses autophagic flux, while gain- and loss-of-function experiments in vitro and a tumor metastasis mouse model confirmed its role in metastasis.\",\n      \"method\": \"siRNA knockdown and overexpression; GFP-LC3B immunofluorescence; electron microscopy of autophagic vesicles; PI3K/AKT/mTOR western blotting; in vivo metastasis mouse model\",\n      \"journal\": \"The international journal of biochemistry & cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — gain- and loss-of-function with multiple orthogonal autophagy detection methods, single lab\",\n      \"pmids\": [\"36758790\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"GPSM1 in macrophages promotes anti-PD-1 resistance in colorectal cancer by driving M2 polarization and tumor microenvironment immunosuppression. The deubiquitinase USP9X stabilizes GPSM1 by preventing K63-polyubiquitination-mediated degradation. Stabilized GPSM1 promotes MEIS3 nuclear translocation, which activates macrophage colony-stimulating factor (CSF1) expression.\",\n      \"method\": \"ChIP-PCR; mass spectrometry; co-immunoprecipitation; single-cell RNA sequencing; multiplex immunofluorescence; orthotopic CRC xenograft model; mass cytometry; flow cytometry\",\n      \"journal\": \"Journal for immunotherapy of cancer\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — ChIP-PCR, mass spectrometry, and co-immunoprecipitation to establish USP9X-GPSM1-MEIS3-CSF1 axis, with in vivo validation; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"40010765\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Myeloid GPSM1 promotes atherosclerosis by sustaining monocyte activation, chemotaxis, and adhesion through the cAMP/PKA/KLF4/PMP22 axis, which activates the p38/ERK MAPK pathway. Myeloid-specific GPSM1 ablation protects against atherosclerosis in Apoe-/- and AAV-PCSK9 mice, while myeloid-restricted GPSM1 overexpression accelerates disease. siRNA-loaded liposome blockade of PMP22 rescues GPSM1-overexpression mice, and a small molecule inhibiting GPSM1 suppresses atherosclerosis in vivo.\",\n      \"method\": \"Myeloid-specific conditional knockout and overexpression in Apoe-/- and AAV-PCSK9 mouse models; monocyte chemotaxis and adhesion assays; cAMP/PKA/KLF4/PMP22 pathway biochemistry; p38/ERK MAPK western blotting; siRNA-loaded liposome PMP22 blockade; small-molecule GPSM1 inhibitor\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO and OE in vivo with defined molecular pathway plus rescue experiment (PMP22 blockade) and pharmacological validation, single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"41296728\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"AGS3 (GPSM1) forms biomolecular condensates (BMCs) under cellular stress (oxidative, pH, thermal). Stress-induced AGS3 BMCs are biochemically distinct from G3BP1 stress granules and Dcp1a P-bodies. Co-expression of Gαi3 (but not DVL2) reduces stress-induced BMC formation. FRAP analysis reveals distinct diffusion kinetics and restricted fluidity within AGS3 BMCs, and stress shifts AGS3 to a membrane pellet fraction.\",\n      \"method\": \"Fluorescence microscopy; FRAP; immunoblotting of fractionated cell lysates; co-expression experiments with Gαi3 and DVL2; stress induction (oxidative, pH, thermal)\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — FRAP and fractionation with multiple stress conditions and Gαi3 rescue, single lab\",\n      \"pmids\": [\"38264908\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AGS3 (GPSM1) interacts with Dishevelled-2 (DVL2), and this interaction is regulated by protein phosphorylation, subcellular distribution, and a cell-surface GPCR. AGS3 signaling influences β-catenin-regulated transcription through the WNT-Frizzled-Dishevelled axis, suggesting integration between Gαi/GPR signaling and WNT pathway.\",\n      \"method\": \"Co-immunoprecipitation; phosphorylation manipulation; GPCR activation; β-catenin transcription reporter assay; subcellular localization imaging\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP interaction and reporter assay for β-catenin transcription, single lab, single paper\",\n      \"pmids\": [\"32737219\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Macrophages lacking Gαi3, AGS3 (GPSM1), or RGS19, or treated with pertussis toxin, show normal levels of basal autophagy, autophagic induction, autophagic flux, autophagic degradation, and anti-autophagic action, indicating that the Gαi3/AGS3/RGS19 pathway does not regulate autophagy in macrophages (negative finding).\",\n      \"method\": \"LC3 processing western blot; LC3 puncta formation assay; long-lived protein degradation assay; Gpsm1-/-, Gnai3-/-, Rgs19-/- bone marrow-derived macrophages; pertussis toxin treatment\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal autophagy assays in knockout macrophages; negative result replicated across multiple genotypes; single lab\",\n      \"pmids\": [\"24312373\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"GPSM1 in CD4+ T cells (especially Treg cells) restricts CD73+CD103+ Treg cell abundance in adipose tissue. GPSM1 deletion in CD4+ T cells or Treg cells increases adipose Treg cells, reduces inflammation, and improves metabolic parameters. Mechanistically, GPSM1 regulates Treg cell abundance via a RHOA-cell stiffness-TAZ axis. Adoptive transfer of GPSM1-deficient Treg cells improves energy expenditure and glucose/lipid metabolism in Rag1-/- mice.\",\n      \"method\": \"Conditional knockout (CD4-Cre and Treg-specific); CD4-T-cell-specific overexpression; flow cytometry (Treg subset analysis); HFD metabolic phenotyping; adoptive transfer into Rag1-/- mice; RHOA/TAZ pathway biochemistry\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-type-specific KO and OE with adoptive transfer rescue and RHOA-TAZ mechanistic pathway, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"42185270\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"GPSM1 (AGS3) is a receptor-independent G-protein signaling modulator whose GoLoco/GPR motifs bind Gαi-GDP free of Gβγ, allowing it to both inhibit Gαi-mediated signaling and release Gβγ for downstream signaling; its subcellular distribution is dynamically regulated by phosphorylation of T602 and interaction with Gαi/o subunits, and it functions in diverse cellular contexts including immune cell chemotaxis (via Gβγ), TGN-to-membrane cargo trafficking of E-cadherin, epidermal spindle orientation (by antagonizing LGN/GPSM2 at the apical cortex), hypothalamic POMC neuron energy balance (via PI3K/AKT/mTOR and autophagy), myeloid cell pro-inflammatory signaling (via Gαi3/cAMP/PKA/CREB/TNFAIP3/NF-κB and cAMP/PKA/KLF4/PMP22/MAPK axes), T-cell/Treg homeostasis (via RHOA-cell stiffness-TAZ), and tumor microenvironment immunosuppression (via USP9X-stabilized GPSM1 driving MEIS3/CSF1 expression).\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"GPSM1 (AGS3) is a receptor-independent activator of G-protein signaling that uses its GoLoco/GPR motifs to bind Gαi/o-GDP, thereby partitioning heterotrimeric G-protein subunits and tuning both Gαi-dependent and Gβγ-dependent outputs across many cell types [#0, #4]. In intact cells GPSM1 assembles with Gαi1 and dynamically dissociates from a cortical signaling complex upon GPCR activation in a pertussis-toxin- and RGS4-sensitive manner [#0], and its subcellular distribution is governed by phosphorylation of a single GPR-domain threonine (T602), with Gαi or Gαo co-expression—but not Gαs or Gαq—redirecting its localization [#4]. Through the released Gβγ arm, GPSM1 acts as a positive regulator of chemokine-receptor signaling, supporting leukocyte chemotaxis, calcium mobilization, and ERK/Akt activation [#3], potentiates polycystin-1/polycystin-2 channel activity to restrain renal cyst progression [#2], and drives compulsive ethanol-seeking behavior [#1]. A recurrent mechanistic theme is GPSM1 control of the cAMP/PKA/CREB axis: in myeloid cells GPSM1 sustains TLR4-induced NF-κB inflammation via Gαi3/cAMP/PKA/CREB-mediated suppression of TNFAIP3, promoting diet-induced metabolic disease [#5], and drives monocyte activation and atherosclerosis through a cAMP/PKA/KLF4/PMP22–p38/ERK MAPK cascade [#16]. GPSM1 also localizes to and organizes trans-Golgi network membranes, where it directs TMED7/E-cadherin cargo trafficking required for early embryonic cell contacts [#6], and it antagonizes its paralog LGN/GPSM2 at the apical cortex to bias epidermal spindle orientation and cell fate [#11]. In immunometabolic and oncogenic settings, GPSM1 modulates PI3K/AKT/mTOR signaling and autophagy in POMC neurons and colorectal cancer cells [#13, #14], shapes Treg homeostasis via a RHOA–cell-stiffness–TAZ axis [#20], and—when stabilized by the deubiquitinase USP9X—promotes MEIS3-driven CSF1 expression and tumor immunosuppression [#15].\",\n  \"teleology\": [\n    {\n      \"year\": 2008,\n      \"claim\": \"Established that GPSM1 acts upstream of Gβγ signaling in a behavioral context, defining its pathway position rather than merely correlating its expression with a phenotype.\",\n      \"evidence\": \"Viral siRNA knockdown in rat nucleus accumbens with operant ethanol self-administration and Gβγ-sequestration epistasis\",\n      \"pmids\": [\"18719114\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not resolve which GPCR engages the GPSM1·Gαi module in this circuit\", \"Molecular partners of the released Gβγ in NAcore neurons not identified\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Showed that a seven-transmembrane receptor directly regulates the GPSM1·Gαi module, answering whether this 'receptor-independent' modulator is itself responsive to GPCR activation in living cells.\",\n      \"evidence\": \"BRET in mammalian cells with α2-adrenergic/μ-opioid receptor activation, pertussis toxin, RGS4, and GRK2-ct controls\",\n      \"pmids\": [\"20716524\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which receptor activation triggers cortical complex dissociation not defined\", \"Stoichiometry of the GPSM1·Gαi·receptor assembly unknown\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Demonstrated a physiological output for GPSM1-released Gβγ on an ion channel, linking the modulator to organ-level disease progression.\",\n      \"evidence\": \"Electrophysiology of polycystin-1/2 channels plus Gpsm1-knockout × Pkd1 hypomorph mouse cross\",\n      \"pmids\": [\"23236168\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct Gβγ–polycystin interaction not structurally defined\", \"Whether GPSM1 acts at the channel locale or upstream is unresolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined GPSM1 as a positive regulator of immune-cell chemokine receptor signaling, clarifying the direction of its effect on G-protein output.\",\n      \"evidence\": \"AGS3-knockout B/T lymphocytes and dendritic cells with chemotaxis, calcium flux, and ERK/Akt assays\",\n      \"pmids\": [\"24573680\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Which chemokine receptors physically couple to GPSM1 not mapped\", \"Relative contribution of Gαi vs Gβγ arms not separated in these cells\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Connected GPSM1 to cAMP/PKA/CREB anti-apoptotic and adhesion signaling in a cancer context, beginning a recurring pathway theme.\",\n      \"evidence\": \"siRNA knockdown in multiple myeloma cells with p-CREB, apoptosis, and fibronectin/HS-5 adhesion assays\",\n      \"pmids\": [\"24307516\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct link from GPSM1/Gαi to adenylyl cyclase not established\", \"Single cell-line system without in vivo validation\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Provided a negative control on the autophagy hypothesis, showing the Gαi3/AGS3/RGS19 module is dispensable for macrophage autophagy under the conditions tested.\",\n      \"evidence\": \"LC3 processing, puncta, and long-lived protein degradation assays in Gpsm1-/-, Gnai3-/-, Rgs19-/- macrophages with pertussis toxin\",\n      \"pmids\": [\"24312373\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Negative result is context-specific to macrophages and may not generalize to other cell types\", \"Does not exclude autophagy roles in non-myeloid cells\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Extended the cAMP/PKA/CREB axis to differentiation control, showing GPSM1 suppresses osteogenic gene programs.\",\n      \"evidence\": \"siRNA knockdown and overexpression in TNF-α-treated dental pulp stem cells with p-CREB and osteogenic marker readouts\",\n      \"pmids\": [\"26143356\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"G-protein step linking GPSM1 to CREB not biochemically dissected\", \"In vivo relevance not tested\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identified a single phosphorylation switch (T602) and Gαi/o binding as the determinants of GPSM1 subcellular distribution, providing a mechanistic basis for its dynamic localization.\",\n      \"evidence\": \"Site-directed mutagenesis (T602A/E/D and reversion), fluorescence microscopy, alkaline phosphatase gel shift, and Gα-subunit co-expression rescue\",\n      \"pmids\": [\"30404823\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Kinase responsible for T602 phosphorylation not identified\", \"Functional consequence of punctate vs cortical distribution for signaling not directly measured\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Placed GPSM1 at the trans-Golgi network controlling E-cadherin cargo trafficking, expanding its role beyond plasma-membrane G-protein signaling to membrane logistics.\",\n      \"evidence\": \"CRISPR/Cas9 knockout in mouse embryos with TGN46/TMED7 live imaging and Gαi1 rescue\",\n      \"pmids\": [\"33148610\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular machinery linking GPSM1·Gαi to TMED7 cargo selection not defined\", \"Whether Gβγ release participates in TGN trafficking unresolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Linked GPSM1's inhibition of Gαi3 to Bcl-2 phosphorylation at TGN-associated membranes, suggesting a route to autophagic signaling regulation.\",\n      \"evidence\": \"Phospho-Bcl-2 immunoblotting with AGS3 overexpression/loss in an RGS4-KO background and adrenal autophagic flux\",\n      \"pmids\": [\"32501280\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"GPSM1 effect inferred indirectly through RGS4 manipulation\", \"Causal chain from phospho-Bcl-2 to autophagy outcome not established\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected GPSM1 to WNT/β-catenin signaling through a phosphorylation- and GPCR-regulated interaction with DVL2.\",\n      \"evidence\": \"Co-immunoprecipitation, phosphorylation manipulation, and β-catenin transcription reporter assay\",\n      \"pmids\": [\"32737219\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single Co-IP without reciprocal structural validation\", \"Direct effect of GPSM1 on Frizzled–Dishevelled assembly not shown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established GPSM1 as a pro-survival regulator in granulosa cells operating through cAMP-PKA-CREB and Bcl-2/Bax balance.\",\n      \"evidence\": \"siRNA knockdown in rat granulosa cells with cAMP measurement, proliferation, apoptosis, and pathway immunoblots\",\n      \"pmids\": [\"33220708\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"G-protein step coupling GPSM1 to cAMP not dissected\", \"Single-species, single-cell-type model\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Tied GPSM1 to a cAMP-generating ADCY6-RAPGEF3-JNK survival pathway in leukemia cells.\",\n      \"evidence\": \"siRNA knockdown in B-ALL cells with proliferation, apoptosis, cell-cycle, and ADCY6/RAPGEF3/JNK immunoblots\",\n      \"pmids\": [\"34257610\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which GPSM1 controls ADCY6 expression unknown\", \"No in vivo confirmation\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined a complete myeloid signaling cascade by which GPSM1 sustains inflammation, answering how it drives metabolic disease at the molecular level.\",\n      \"evidence\": \"Myeloid-specific conditional knockout mice on HFD with Gαi3/cAMP/PKA/CREB/TNFAIP3/NF-κB biochemistry and small-molecule inhibitor\",\n      \"pmids\": [\"36434066\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream receptor coupling GPSM1 to Gαi3 in macrophages not identified\", \"Transcriptional mechanism of CREB-mediated TNFAIP3 repression not fully mapped\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed GPSM1 antagonizes its paralog LGN/GPSM2 at the apical cortex to set spindle orientation and cell fate, defining a paralog-competition mechanism.\",\n      \"evidence\": \"Conditional knockout/overexpression in mouse epidermis with live imaging, double-mutant epistasis, and clonal lineage tracing\",\n      \"pmids\": [\"37017303\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of cortical LGN displacement by GPSM1 not resolved\", \"Role of Gαi in this cortical competition not directly tested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Established a neuronal role for GPSM1 in energy balance via PI3K/AKT/mTOR-dependent autophagy and leptin sensitivity in POMC neurons.\",\n      \"evidence\": \"POMC-specific conditional knockout mice with HFD phenotyping, autophagy assays, and sympathetic innervation/BAT thermogenesis analysis\",\n      \"pmids\": [\"37979657\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How GPSM1 G-protein activity feeds into PI3K/AKT/mTOR not defined\", \"Direct substrate or effector connecting GPSM1 to autophagy machinery unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed GPSM1 suppresses autophagic flux and promotes metastasis in colorectal cancer through PI3K/AKT/mTOR activation.\",\n      \"evidence\": \"Gain/loss-of-function with GFP-LC3B imaging, autophagic vesicle electron microscopy, pathway immunoblots, and a metastasis mouse model\",\n      \"pmids\": [\"36758790\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanistic link from GPSM1 to PI3K activation not defined\", \"Whether autophagy suppression is causal for metastasis not separated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed that GPSM1 forms stress-induced biomolecular condensates distinct from stress granules and P-bodies, adding a phase-separation behavior regulated by Gαi3.\",\n      \"evidence\": \"Fluorescence microscopy, FRAP, lysate fractionation, and G αi3/DVL2 co-expression under oxidative/pH/thermal stress\",\n      \"pmids\": [\"38264908\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional role of GPSM1 condensates in signaling or stress response unknown\", \"Composition of the condensates not characterized\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined a deubiquitination-driven oncogenic axis in which USP9X stabilizes GPSM1 to promote MEIS3/CSF1-mediated tumor immunosuppression and immunotherapy resistance.\",\n      \"evidence\": \"Mass spectrometry, co-immunoprecipitation, ChIP-PCR, single-cell RNA-seq, and orthotopic CRC xenografts with mass/flow cytometry\",\n      \"pmids\": [\"40010765\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How GPSM1 promotes MEIS3 nuclear translocation mechanistically not resolved\", \"Whether G-protein modulator activity is required for the MEIS3/CSF1 output unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified a complete myeloid cAMP/PKA/KLF4/PMP22–MAPK cascade through which GPSM1 drives atherosclerosis, and validated it pharmacologically and by rescue.\",\n      \"evidence\": \"Myeloid conditional KO and overexpression in Apoe-/- and AAV-PCSK9 mice with chemotaxis/adhesion assays, pathway biochemistry, PMP22 siRNA-liposome rescue, and a small-molecule GPSM1 inhibitor\",\n      \"pmids\": [\"41296728\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream receptor/Gα coupling in monocytes not defined\", \"Relationship between this KLF4/PMP22 cascade and the TNFAIP3/NF-κB cascade in myeloid cells not reconciled\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Established GPSM1 as a restrictor of adipose Treg homeostasis acting through a RHOA–cell-stiffness–TAZ mechanotransduction axis, with metabolic consequences confirmed by adoptive transfer.\",\n      \"evidence\": \"CD4/Treg-specific conditional KO and overexpression with HFD phenotyping, Treg subset flow cytometry, RHOA/TAZ biochemistry, and adoptive transfer into Rag1-/- mice\",\n      \"pmids\": [\"42185270\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How GPSM1 G-protein activity engages RHOA/cell stiffness not defined\", \"Direct GPSM1 effectors in Treg cells not identified\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The unifying question remains how a single GoLoco/GPR Gαi/o modulator selects among its many divergent outputs—Gβγ-driven chemotaxis and channel modulation, cAMP/PKA/CREB inflammation, PI3K/AKT/mTOR autophagy, TGN cargo trafficking, cortical LGN antagonism, and ubiquitin-stabilized transcriptional programs—and which upstream receptors and cell-type-specific cofactors dictate this choice.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model reconciling Gαi-binding with the diverse downstream cascades\", \"Upstream GPCRs coupling GPSM1 in most contexts unidentified\", \"Determinants of context-specific pathway selection unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 3, 4, 5]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 2, 3]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 4, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [4, 17]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 6, 11]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [6, 7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 3, 4, 5]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [3, 5, 15, 20]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [13, 14]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [6, 11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"GNAI1\", \"GNAI3\", \"GNAO1\", \"DVL2\", \"GPSM2\", \"USP9X\", \"RGS4\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}