{"gene":"DGKA","run_date":"2026-04-28T17:46:02","timeline":{"discoveries":[{"year":2012,"finding":"DGK-α (DGKA) expression is elevated in human CD8+ tumor-infiltrating T cells and suppresses distal TCR signaling (ERK, JNK, AKT phosphorylation); pharmacologic inhibition of DGK-α restored ERK phosphorylation and lytic granule exocytosis, placing DGK-α downstream of PLC-γ but upstream of RAS/MAPK and PI3K/AKT in the TCR signaling cascade.","method":"Pharmacologic inhibition of DGK-α in primary human CD8+ TILs; western blot for phospho-ERK/JNK/AKT; degranulation assay; IL-2-mediated reduction of DGK-α protein levels","journal":"Journal of Immunology","confidence":"High","confidence_rationale":"Tier 2 — clean pharmacologic KD with defined signaling phenotype, replicated with IL-2 modulation; multiple orthogonal readouts","pmids":["22573804"],"is_preprint":false},{"year":2020,"finding":"DGKA confers platinum resistance in ovarian cancer through production of phosphatidic acid (PA), which facilitates JNK recruitment to c-JUN and its nuclear localization, leading to c-JUN activation and consequent transcriptional upregulation of the cell-cycle regulator WEE1.","method":"RNAi synthetic lethal screening; DGKA kinase-dead variants; phospho-proteomic and genomic screens; pharmacologic inhibition; patient-derived tumor validation","journal":"Clinical Cancer Research","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (genetic variants, proteomics, pharmacology, patient samples) in a single study establishing a defined signaling pathway","pmids":["32341033"],"is_preprint":false},{"year":2022,"finding":"DGKA directly interacts with SRC (via the SRC SH3 domain binding to DGKA's C-terminal domain) and FAK (via DGKA's catalytic domain binding to FAK's FREM domain); DGKA phosphorylates SRC at Tyr416 and FAK at Tyr397 to form and activate the DGKA/SRC/FAK complex, which initiates downstream WNT/β-catenin and VEGF signaling, promoting EMT and angiogenesis in NSCLC.","method":"Co-immunoprecipitation; domain-mapping experiments with truncation constructs; western blot for phospho-SRC(Y416) and phospho-FAK(Y397); DGKA knockdown in vitro invasion assays; in vivo pharmacologic inhibition","journal":"Cancer Letters","confidence":"Medium","confidence_rationale":"Tier 2/3 — reciprocal Co-IP with domain mapping and phosphorylation readouts, single lab study","pmids":["35131384"],"is_preprint":false},{"year":2021,"finding":"DGKA mediates resistance to anti-PD-1 therapy by exacerbating exhaustion of reinvigorated tumor-specific T cells; pharmacologic ablation of DGKA postponed T-cell exhaustion. Additionally, DGKA expression in cancer cells promotes tumor growth via the AKT signaling pathway.","method":"Pharmacologic inhibition of DGKA in vivo tumor models; T-cell exhaustion markers; AKT pathway western blotting","journal":"Cancer Immunology Research","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo genetic/pharmacologic model with defined T-cell exhaustion phenotype and AKT pathway readout, single lab","pmids":["33608256"],"is_preprint":false},{"year":2021,"finding":"DGKA supports macropinocytosis in TSC2-deficient cells to maintain phospholipid homeostasis; phosphatidic acid levels were 5-fold elevated in Tsc2-/- MEFs, and DGKA inhibition (ritanserin) depleted PA, reduced macropinocytic albumin uptake, decreased lysosome number and activity, and prevented alveolar destruction in a mouse LAM model.","method":"High-throughput drug screen; PA lipidomic quantification; macropinocytosis assay; lysosome quantification; Tsc2-/- MEF genetic model; in vivo mouse LAM model with genetic DGKA knockdown","journal":"Cancer Research","confidence":"High","confidence_rationale":"Tier 1–2 — lipidomic quantification, genetic KO, in vivo validation, multiple orthogonal methods establishing DGKA-PA-macropinocytosis axis","pmids":["33593821"],"is_preprint":false},{"year":2020,"finding":"DGKα and DGKζ activities control TH1 and TH17 cell differentiation downstream of TCR signaling; single deficiency of DGKα impairs TH1 but not TH2/TH17 differentiation, whereas double ablation of DGKα and DGKζ promotes both TH1 and TH17 differentiation, with dysregulated TH17 differentiation driven by increased mTORC1/S6K1 signaling.","method":"DGKα and DGKζ single and double knockout mouse T cells; in vitro TH differentiation assays; in vivo airway inflammation model; mTORC1/S6K1 pathway western blotting","journal":"Frontiers in Immunology","confidence":"Medium","confidence_rationale":"Tier 2 — genetic KO with defined differentiation phenotype and pathway placement, single lab","pmids":["32010133"],"is_preprint":false},{"year":2013,"finding":"DGK-α is a direct target of miR-297 in glioblastoma; miR-297 overexpression reduced DGK-α protein levels (confirmed by 3'UTR luciferase assay), and rescue with DGK-α overexpression reversed miR-297-induced cytotoxicity, demonstrating DGK-α is a critical survival factor downstream of this miRNA axis. Hypoxia and hnRNPL upregulate DGK-α and buffer miR-297 cytotoxic effects.","method":"3'UTR luciferase reporter assay; immunoblot; DGK-α overexpression rescue; glioblastoma xenograft model; hypoxia treatment","journal":"Neuro-oncology","confidence":"Medium","confidence_rationale":"Tier 2/3 — 3'UTR validation plus rescue experiment confirms direct miR-297 target relationship, single lab","pmids":["24158111"],"is_preprint":false},{"year":2023,"finding":"DGKA promotes intrahepatic cholangiocarcinoma cell proliferation by enhancing phosphatidic acid (PA) metabolism and activating MAPK signaling.","method":"Cell proliferation assay; EdU assay; colony formation assay; RNA-sequencing; western blotting; DGKA knockdown/overexpression","journal":"Cancer Communications","confidence":"Medium","confidence_rationale":"Tier 2/3 — multiple cellular assays with pathway readout (MAPK), single lab","pmids":["38143235"],"is_preprint":false},{"year":2025,"finding":"A dual DGKα/ζ inhibitor (INCB165451) enhances human TIL anti-tumor efficacy by increasing intratumoral T-cell infiltration, upregulating inflammatory gene expression, and reducing TIL hypofunction (enhanced cytokine production), confirming that DGKα negatively regulates T cells through the DAG signaling pathway.","method":"NSCLC mouse adoptive transfer model; ex vivo human TIL stimulation; gene/protein expression of cytokines/chemokines; precision-cut tumor slice assay with BiTE stimulation","journal":"Oncoimmunology","confidence":"Medium","confidence_rationale":"Tier 2 — dual inhibitor with multiple in vivo and ex vivo human readouts, single lab, no isoform-specific separation","pmids":["41459935"],"is_preprint":false},{"year":2018,"finding":"In Dictyostelium discoideum, loss of the single DGKA gene reduces sensitivity to valproic acid and lithium, and DGKA is necessary for compound-specific increases in DAG levels following VPA or lithium treatment, placing DGKA in the phosphoinositide recycling/DAG regulation pathway relevant to seizure and bipolar disorder signaling.","method":"dgkA null mutant phenotypic analysis; DAG level measurement; drug sensitivity assays with VPA, lithium, and branched fatty acids","journal":"Disease Models & Mechanisms","confidence":"Low","confidence_rationale":"Tier 3 — Dictyostelium model organism (not an ortholog of human DGKA by domain architecture), single lab","pmids":["30135067"],"is_preprint":false}],"current_model":"DGKA (diacylglycerol kinase alpha) is a lipid kinase that phosphorylates diacylglycerol (DAG) to produce phosphatidic acid (PA), thereby attenuating DAG-dependent signaling; in T cells it acts downstream of PLC-γ to suppress RAS/MAPK (ERK/JNK) and PI3K/AKT activation, restraining T-cell effector function and promoting exhaustion, while in tumor cells PA produced by DGKA activates JNK-c-JUN-WEE1 signaling for chemoresistance, directly activates and scaffolds an SRC/FAK complex via domain-specific interactions to drive EMT and metastasis, sustains macropinocytosis-dependent phospholipid homeostasis in mTORC1-hyperactive cells, and activates MAPK signaling to promote proliferation."},"narrative":{"teleology":[{"year":2012,"claim":"Establishing DGKA as a checkpoint in TCR signaling that suppresses effector function in human TILs resolved how DAG metabolism restrains anti-tumor immunity downstream of PLC-γ.","evidence":"Pharmacologic DGK-α inhibition in primary human CD8+ TILs with phospho-ERK/JNK/AKT western blots and degranulation assays","pmids":["22573804"],"confidence":"High","gaps":["Relative contributions of DGKA versus DGKζ in TIL hypofunction were not separated","No genetic knockout confirmation in human T cells","Whether DGKA inhibition affects T-cell persistence in vivo was untested"]},{"year":2013,"claim":"Identifying DGKA as a direct miR-297 target and survival factor in glioblastoma showed that post-transcriptional regulation of DGKA levels determines tumor cell viability under hypoxic stress.","evidence":"3′UTR luciferase reporter assay and DGKA overexpression rescue of miR-297-induced cytotoxicity in glioblastoma cells","pmids":["24158111"],"confidence":"Medium","gaps":["No endogenous miR-297 loss-of-function to confirm physiological regulation","Downstream effectors of DGKA-dependent survival were not mapped"]},{"year":2020,"claim":"Demonstrating that DGKA-produced PA recruits JNK to c-JUN, driving WEE1 transcription and platinum resistance, defined a complete PA → JNK → c-JUN → WEE1 signaling axis in ovarian cancer chemoresistance.","evidence":"RNAi synthetic lethal screen, kinase-dead DGKA mutants, phosphoproteomics, pharmacologic inhibition, and patient-derived tumor validation","pmids":["32341033"],"confidence":"High","gaps":["Direct PA-JNK binding site or mechanism of recruitment not structurally resolved","Whether this axis operates in other platinum-treated tumor types was untested"]},{"year":2020,"claim":"Genetic ablation studies separating DGKα and DGKζ contributions showed that DGKα specifically promotes TH1 differentiation while double deficiency unleashes TH17 via mTORC1/S6K1, revealing isoform-specific control of T-helper fate.","evidence":"DGKα and DGKζ single and double knockout mouse T cells with in vitro TH differentiation assays and mTORC1/S6K1 western blotting","pmids":["32010133"],"confidence":"Medium","gaps":["Mechanism by which DGKα selectively supports TH1 over TH17 not identified","Human T-helper differentiation effects not confirmed"]},{"year":2021,"claim":"Showing that DGKA sustains macropinocytosis and phospholipid homeostasis in TSC2-deficient cells via a PA-dependent axis linked lipid kinase activity to nutrient acquisition in mTORC1-hyperactive tumors.","evidence":"PA lipidomics in Tsc2−/− MEFs, macropinocytosis assays, lysosome quantification, genetic DGKA knockdown in a mouse LAM model","pmids":["33593821"],"confidence":"High","gaps":["Whether DGKA-PA drives macropinocytosis in other mTORC1-hyperactive contexts (e.g., PTEN-loss) was not tested","PA effector(s) that directly regulate macropinosome formation remain unidentified"]},{"year":2021,"claim":"Demonstrating that DGKA inhibition delays exhaustion of reinvigorated T cells during anti-PD-1 therapy established DGKA as a combinatorial immunotherapy target beyond simple T-cell activation.","evidence":"In vivo tumor models with pharmacologic DGKA inhibition combined with anti-PD-1; T-cell exhaustion markers and AKT pathway analysis","pmids":["33608256"],"confidence":"Medium","gaps":["Molecular mechanism by which DGKA promotes exhaustion versus hypofunction was not delineated","Cancer cell-intrinsic AKT effects were not fully separated from T-cell effects"]},{"year":2022,"claim":"Domain-mapping of DGKA interactions with SRC (via SH3–C-terminal domain) and FAK (via catalytic domain–FERM domain) revealed an unexpected scaffolding and tyrosine kinase-like activity, expanding DGKA function beyond lipid phosphorylation to direct protein phosphorylation and EMT/angiogenesis signaling.","evidence":"Reciprocal Co-IP with truncation constructs, phospho-SRC(Y416)/phospho-FAK(Y397) western blots, in vitro invasion assays and in vivo pharmacologic inhibition in NSCLC","pmids":["35131384"],"confidence":"Medium","gaps":["Direct in vitro kinase assay demonstrating DGKA phosphorylates SRC/FAK as a protein kinase was not shown","Structural basis for a lipid kinase phosphorylating tyrosine residues is unresolved","Single-lab finding; independent replication needed"]},{"year":2023,"claim":"Confirmation that DGKA promotes proliferation through PA-MAPK signaling in cholangiocarcinoma generalized the PA → MAPK proliferative axis beyond ovarian cancer.","evidence":"DGKA knockdown/overexpression with EdU, colony formation, RNA-seq, and MAPK western blotting in cholangiocarcinoma cells","pmids":["38143235"],"confidence":"Medium","gaps":["Specific MAPK components activated by PA were not mapped","In vivo validation in cholangiocarcinoma models was limited"]},{"year":2025,"claim":"A dual DGKα/ζ inhibitor validated in human TIL systems confirmed clinical translational potential of DGKA inhibition to reverse T-cell hypofunction, though isoform-specific contributions could not be separated.","evidence":"NSCLC mouse adoptive transfer, ex vivo human TIL stimulation, precision-cut tumor slice assay with BiTE","pmids":["41459935"],"confidence":"Medium","gaps":["Isoform-specific contribution of DGKα versus DGKζ not deconvolved","Long-term in vivo efficacy and autoimmune toxicity not assessed"]},{"year":null,"claim":"The structural basis for DGKA's reported protein kinase activity toward SRC and FAK, the identity of direct PA effectors mediating macropinocytosis, and full isoform-specific separation of DGKα versus DGKζ in human anti-tumor immunity remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No crystal or cryo-EM structure of DGKA to explain dual lipid/protein kinase activity","Direct PA-binding effectors in macropinocytosis not identified","Isoform-selective DGKα inhibitors for clean human in vivo studies are lacking"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,1,2,4,7]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[1,4]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,3,5]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[0,3,5,8]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[1,4,7]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[1,3,6]}],"complexes":["DGKA/SRC/FAK complex"],"partners":["SRC","FAK","JNK","C-JUN","WEE1"],"other_free_text":[]},"mechanistic_narrative":"DGKA (diacylglycerol kinase alpha) is a lipid kinase that phosphorylates diacylglycerol (DAG) to phosphatidic acid (PA), functioning as a critical signaling switch that terminates DAG-dependent pathways while simultaneously generating PA as a bioactive second messenger. In T cells, DGKA acts downstream of PLC-γ to suppress RAS/MAPK (ERK/JNK), PI3K/AKT, and mTORC1 signaling, restraining effector function and promoting T-cell exhaustion; its inhibition restores cytolytic capacity of tumor-infiltrating lymphocytes and enhances anti-PD-1 immunotherapy responses [PMID:22573804, PMID:33608256, PMID:41459935]. In cancer cells, DGKA-generated PA drives distinct oncogenic programs: it facilitates JNK-mediated c-JUN activation and WEE1 upregulation to confer platinum chemoresistance, activates MAPK signaling for proliferation, and sustains macropinocytosis-dependent phospholipid homeostasis in mTORC1-hyperactive cells [PMID:32341033, PMID:33593821, PMID:38143235]. DGKA also directly binds and phosphorylates SRC (Tyr416) and FAK (Tyr397) through domain-specific interactions, scaffolding a DGKA/SRC/FAK complex that promotes epithelial–mesenchymal transition and angiogenesis via WNT/β-catenin and VEGF signaling [PMID:35131384]."},"prefetch_data":{"uniprot":{"accession":"P23743","full_name":"Diacylglycerol kinase alpha","aliases":["80 kDa diacylglycerol kinase","Diglyceride kinase alpha","DGK-alpha"],"length_aa":735,"mass_kda":82.6,"function":"Diacylglycerol kinase that converts diacylglycerol/DAG into phosphatidic acid/phosphatidate/PA and regulates the respective levels of these two bioactive lipids (PubMed:15544348, PubMed:2175712). Thereby, acts as a central switch between the signaling pathways activated by these second messengers with different cellular targets and opposite effects in numerous biological processes (PubMed:15544348, PubMed:2175712). Also plays an important role in the biosynthesis of complex lipids (Probable). Can also phosphorylate 1-alkyl-2-acylglycerol in vitro as efficiently as diacylglycerol provided it contains an arachidonoyl group (PubMed:15544348). Also involved in the production of alkyl-lysophosphatidic acid, another bioactive lipid, through the phosphorylation of 1-alkyl-2-acetyl glycerol (PubMed:22627129)","subcellular_location":"Cytoplasm, cytosol","url":"https://www.uniprot.org/uniprotkb/P23743/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DGKA","classification":"Not Classified","n_dependent_lines":8,"n_total_lines":1208,"dependency_fraction":0.006622516556291391},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/DGKA","total_profiled":1310},"omim":[{"mim_id":"616711","title":"TAO KINASE 3; TAOK3","url":"https://www.omim.org/entry/616711"},{"mim_id":"610681","title":"PHOSPHOFRUCTOKINASE, MUSCLE TYPE; PFKM","url":"https://www.omim.org/entry/610681"},{"mim_id":"610307","title":"CERAMIDE KINASE; CERK","url":"https://www.omim.org/entry/610307"},{"mim_id":"607092","title":"SPHINGOSINE KINASE 2; SPHK2","url":"https://www.omim.org/entry/607092"},{"mim_id":"125855","title":"DIACYLGLYCEROL KINASE, ALPHA, 80-KD; DGKA","url":"https://www.omim.org/entry/125855"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Mitochondria","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"lymphoid tissue","ntpm":184.5}],"url":"https://www.proteinatlas.org/search/DGKA"},"hgnc":{"alias_symbol":["DGK-alpha"],"prev_symbol":["DAGK","DAGK1"]},"alphafold":{"accession":"P23743","domains":[{"cath_id":"1.10.238.110","chopping":"9-193","consensus_level":"medium","plddt":74.163,"start":9,"end":193},{"cath_id":"3.40.50.10330","chopping":"365-491","consensus_level":"high","plddt":92.863,"start":365,"end":491},{"cath_id":"2.60.200.40","chopping":"499-717","consensus_level":"high","plddt":89.8995,"start":499,"end":717}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P23743","model_url":"https://alphafold.ebi.ac.uk/files/AF-P23743-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P23743-F1-predicted_aligned_error_v6.png","plddt_mean":82.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DGKA","jax_strain_url":"https://www.jax.org/strain/search?query=DGKA"},"sequence":{"accession":"P23743","fasta_url":"https://rest.uniprot.org/uniprotkb/P23743.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P23743/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P23743"}},"corpus_meta":[{"pmid":"22573804","id":"PMC_22573804","title":"High DGK-α and disabled MAPK pathways cause dysfunction of human tumor-infiltrating CD8+ T cells that is reversible by pharmacologic intervention.","date":"2012","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/22573804","citation_count":77,"is_preprint":false},{"pmid":"32341033","id":"PMC_32341033","title":"DGKA Provides Platinum Resistance in Ovarian Cancer Through Activation of c-JUN-WEE1 Signaling.","date":"2020","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/32341033","citation_count":53,"is_preprint":false},{"pmid":"24158111","id":"PMC_24158111","title":"A miR-297/hypoxia/DGK-α axis regulating glioblastoma survival.","date":"2013","source":"Neuro-oncology","url":"https://pubmed.ncbi.nlm.nih.gov/24158111","citation_count":40,"is_preprint":false},{"pmid":"21463370","id":"PMC_21463370","title":"Antagonistic regulation of dgkA and plsB genes of phospholipid synthesis by multiple stress responses in Escherichia coli.","date":"2011","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/21463370","citation_count":36,"is_preprint":false},{"pmid":"28316970","id":"PMC_28316970","title":"DGK-α: A Checkpoint in Cancer-Mediated Immuno-Inhibition and Target for Immunotherapy.","date":"2017","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/28316970","citation_count":35,"is_preprint":false},{"pmid":"35131384","id":"PMC_35131384","title":"DGKA interacts with SRC/FAK to promote the metastasis of non-small cell lung cancer.","date":"2022","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/35131384","citation_count":27,"is_preprint":false},{"pmid":"33608256","id":"PMC_33608256","title":"DGKA Mediates Resistance to PD-1 Blockade.","date":"2021","source":"Cancer immunology 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Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/32842668","citation_count":14,"is_preprint":false},{"pmid":"38143235","id":"PMC_38143235","title":"Multiomics analysis reveals metabolic subtypes and identifies diacylglycerol kinase α (DGKA) as a potential therapeutic target for intrahepatic cholangiocarcinoma.","date":"2023","source":"Cancer communications (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/38143235","citation_count":12,"is_preprint":false},{"pmid":"32010133","id":"PMC_32010133","title":"DGK α and ζ Activities Control TH1 and TH17 Cell Differentiation.","date":"2020","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/32010133","citation_count":12,"is_preprint":false},{"pmid":"33593821","id":"PMC_33593821","title":"Therapeutic Targeting of DGKA-Mediated Macropinocytosis Leads to Phospholipid Reprogramming in Tuberous Sclerosis Complex.","date":"2021","source":"Cancer 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research","url":"https://pubmed.ncbi.nlm.nih.gov/24950064","citation_count":6,"is_preprint":false},{"pmid":"41459935","id":"PMC_41459935","title":"A dual diacylglycerol kinase (DGK) alpha/zeta inhibitor augments the activity of human tumor infiltrating lymphocytes in in vivo and ex vivo models.","date":"2025","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/41459935","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10078,"output_tokens":2690,"usd":0.035292},"stage2":{"model":"claude-opus-4-6","input_tokens":6108,"output_tokens":5611,"usd":0.256223},"total_usd":0.291515,"stage1_batch_id":"msgbatch_0118RScg3WGaGkaZRtWZbtce","stage2_batch_id":"msgbatch_01Cpfzvtbj42aqp5oYZ5Te2s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2012,\n      \"finding\": \"DGK-α (DGKA) expression is elevated in human CD8+ tumor-infiltrating T cells and suppresses distal TCR signaling (ERK, JNK, AKT phosphorylation); pharmacologic inhibition of DGK-α restored ERK phosphorylation and lytic granule exocytosis, placing DGK-α downstream of PLC-γ but upstream of RAS/MAPK and PI3K/AKT in the TCR signaling cascade.\",\n      \"method\": \"Pharmacologic inhibition of DGK-α in primary human CD8+ TILs; western blot for phospho-ERK/JNK/AKT; degranulation assay; IL-2-mediated reduction of DGK-α protein levels\",\n      \"journal\": \"Journal of Immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean pharmacologic KD with defined signaling phenotype, replicated with IL-2 modulation; multiple orthogonal readouts\",\n      \"pmids\": [\"22573804\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DGKA confers platinum resistance in ovarian cancer through production of phosphatidic acid (PA), which facilitates JNK recruitment to c-JUN and its nuclear localization, leading to c-JUN activation and consequent transcriptional upregulation of the cell-cycle regulator WEE1.\",\n      \"method\": \"RNAi synthetic lethal screening; DGKA kinase-dead variants; phospho-proteomic and genomic screens; pharmacologic inhibition; patient-derived tumor validation\",\n      \"journal\": \"Clinical Cancer Research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (genetic variants, proteomics, pharmacology, patient samples) in a single study establishing a defined signaling pathway\",\n      \"pmids\": [\"32341033\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"DGKA directly interacts with SRC (via the SRC SH3 domain binding to DGKA's C-terminal domain) and FAK (via DGKA's catalytic domain binding to FAK's FREM domain); DGKA phosphorylates SRC at Tyr416 and FAK at Tyr397 to form and activate the DGKA/SRC/FAK complex, which initiates downstream WNT/β-catenin and VEGF signaling, promoting EMT and angiogenesis in NSCLC.\",\n      \"method\": \"Co-immunoprecipitation; domain-mapping experiments with truncation constructs; western blot for phospho-SRC(Y416) and phospho-FAK(Y397); DGKA knockdown in vitro invasion assays; in vivo pharmacologic inhibition\",\n      \"journal\": \"Cancer Letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2/3 — reciprocal Co-IP with domain mapping and phosphorylation readouts, single lab study\",\n      \"pmids\": [\"35131384\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DGKA mediates resistance to anti-PD-1 therapy by exacerbating exhaustion of reinvigorated tumor-specific T cells; pharmacologic ablation of DGKA postponed T-cell exhaustion. Additionally, DGKA expression in cancer cells promotes tumor growth via the AKT signaling pathway.\",\n      \"method\": \"Pharmacologic inhibition of DGKA in vivo tumor models; T-cell exhaustion markers; AKT pathway western blotting\",\n      \"journal\": \"Cancer Immunology Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic/pharmacologic model with defined T-cell exhaustion phenotype and AKT pathway readout, single lab\",\n      \"pmids\": [\"33608256\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DGKA supports macropinocytosis in TSC2-deficient cells to maintain phospholipid homeostasis; phosphatidic acid levels were 5-fold elevated in Tsc2-/- MEFs, and DGKA inhibition (ritanserin) depleted PA, reduced macropinocytic albumin uptake, decreased lysosome number and activity, and prevented alveolar destruction in a mouse LAM model.\",\n      \"method\": \"High-throughput drug screen; PA lipidomic quantification; macropinocytosis assay; lysosome quantification; Tsc2-/- MEF genetic model; in vivo mouse LAM model with genetic DGKA knockdown\",\n      \"journal\": \"Cancer Research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — lipidomic quantification, genetic KO, in vivo validation, multiple orthogonal methods establishing DGKA-PA-macropinocytosis axis\",\n      \"pmids\": [\"33593821\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DGKα and DGKζ activities control TH1 and TH17 cell differentiation downstream of TCR signaling; single deficiency of DGKα impairs TH1 but not TH2/TH17 differentiation, whereas double ablation of DGKα and DGKζ promotes both TH1 and TH17 differentiation, with dysregulated TH17 differentiation driven by increased mTORC1/S6K1 signaling.\",\n      \"method\": \"DGKα and DGKζ single and double knockout mouse T cells; in vitro TH differentiation assays; in vivo airway inflammation model; mTORC1/S6K1 pathway western blotting\",\n      \"journal\": \"Frontiers in Immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with defined differentiation phenotype and pathway placement, single lab\",\n      \"pmids\": [\"32010133\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"DGK-α is a direct target of miR-297 in glioblastoma; miR-297 overexpression reduced DGK-α protein levels (confirmed by 3'UTR luciferase assay), and rescue with DGK-α overexpression reversed miR-297-induced cytotoxicity, demonstrating DGK-α is a critical survival factor downstream of this miRNA axis. Hypoxia and hnRNPL upregulate DGK-α and buffer miR-297 cytotoxic effects.\",\n      \"method\": \"3'UTR luciferase reporter assay; immunoblot; DGK-α overexpression rescue; glioblastoma xenograft model; hypoxia treatment\",\n      \"journal\": \"Neuro-oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2/3 — 3'UTR validation plus rescue experiment confirms direct miR-297 target relationship, single lab\",\n      \"pmids\": [\"24158111\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"DGKA promotes intrahepatic cholangiocarcinoma cell proliferation by enhancing phosphatidic acid (PA) metabolism and activating MAPK signaling.\",\n      \"method\": \"Cell proliferation assay; EdU assay; colony formation assay; RNA-sequencing; western blotting; DGKA knockdown/overexpression\",\n      \"journal\": \"Cancer Communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2/3 — multiple cellular assays with pathway readout (MAPK), single lab\",\n      \"pmids\": [\"38143235\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"A dual DGKα/ζ inhibitor (INCB165451) enhances human TIL anti-tumor efficacy by increasing intratumoral T-cell infiltration, upregulating inflammatory gene expression, and reducing TIL hypofunction (enhanced cytokine production), confirming that DGKα negatively regulates T cells through the DAG signaling pathway.\",\n      \"method\": \"NSCLC mouse adoptive transfer model; ex vivo human TIL stimulation; gene/protein expression of cytokines/chemokines; precision-cut tumor slice assay with BiTE stimulation\",\n      \"journal\": \"Oncoimmunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — dual inhibitor with multiple in vivo and ex vivo human readouts, single lab, no isoform-specific separation\",\n      \"pmids\": [\"41459935\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"In Dictyostelium discoideum, loss of the single DGKA gene reduces sensitivity to valproic acid and lithium, and DGKA is necessary for compound-specific increases in DAG levels following VPA or lithium treatment, placing DGKA in the phosphoinositide recycling/DAG regulation pathway relevant to seizure and bipolar disorder signaling.\",\n      \"method\": \"dgkA null mutant phenotypic analysis; DAG level measurement; drug sensitivity assays with VPA, lithium, and branched fatty acids\",\n      \"journal\": \"Disease Models & Mechanisms\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — Dictyostelium model organism (not an ortholog of human DGKA by domain architecture), single lab\",\n      \"pmids\": [\"30135067\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DGKA (diacylglycerol kinase alpha) is a lipid kinase that phosphorylates diacylglycerol (DAG) to produce phosphatidic acid (PA), thereby attenuating DAG-dependent signaling; in T cells it acts downstream of PLC-γ to suppress RAS/MAPK (ERK/JNK) and PI3K/AKT activation, restraining T-cell effector function and promoting exhaustion, while in tumor cells PA produced by DGKA activates JNK-c-JUN-WEE1 signaling for chemoresistance, directly activates and scaffolds an SRC/FAK complex via domain-specific interactions to drive EMT and metastasis, sustains macropinocytosis-dependent phospholipid homeostasis in mTORC1-hyperactive cells, and activates MAPK signaling to promote proliferation.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"DGKA (diacylglycerol kinase alpha) is a lipid kinase that phosphorylates diacylglycerol (DAG) to phosphatidic acid (PA), functioning as a critical signaling switch that terminates DAG-dependent pathways while simultaneously generating PA as a bioactive second messenger. In T cells, DGKA acts downstream of PLC-γ to suppress RAS/MAPK (ERK/JNK), PI3K/AKT, and mTORC1 signaling, restraining effector function and promoting T-cell exhaustion; its inhibition restores cytolytic capacity of tumor-infiltrating lymphocytes and enhances anti-PD-1 immunotherapy responses [PMID:22573804, PMID:33608256, PMID:41459935]. In cancer cells, DGKA-generated PA drives distinct oncogenic programs: it facilitates JNK-mediated c-JUN activation and WEE1 upregulation to confer platinum chemoresistance, activates MAPK signaling for proliferation, and sustains macropinocytosis-dependent phospholipid homeostasis in mTORC1-hyperactive cells [PMID:32341033, PMID:33593821, PMID:38143235]. DGKA also directly binds and phosphorylates SRC (Tyr416) and FAK (Tyr397) through domain-specific interactions, scaffolding a DGKA/SRC/FAK complex that promotes epithelial–mesenchymal transition and angiogenesis via WNT/β-catenin and VEGF signaling [PMID:35131384].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Establishing DGKA as a checkpoint in TCR signaling that suppresses effector function in human TILs resolved how DAG metabolism restrains anti-tumor immunity downstream of PLC-γ.\",\n      \"evidence\": \"Pharmacologic DGK-α inhibition in primary human CD8+ TILs with phospho-ERK/JNK/AKT western blots and degranulation assays\",\n      \"pmids\": [\"22573804\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Relative contributions of DGKA versus DGKζ in TIL hypofunction were not separated\",\n        \"No genetic knockout confirmation in human T cells\",\n        \"Whether DGKA inhibition affects T-cell persistence in vivo was untested\"\n      ]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identifying DGKA as a direct miR-297 target and survival factor in glioblastoma showed that post-transcriptional regulation of DGKA levels determines tumor cell viability under hypoxic stress.\",\n      \"evidence\": \"3′UTR luciferase reporter assay and DGKA overexpression rescue of miR-297-induced cytotoxicity in glioblastoma cells\",\n      \"pmids\": [\"24158111\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No endogenous miR-297 loss-of-function to confirm physiological regulation\",\n        \"Downstream effectors of DGKA-dependent survival were not mapped\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Demonstrating that DGKA-produced PA recruits JNK to c-JUN, driving WEE1 transcription and platinum resistance, defined a complete PA → JNK → c-JUN → WEE1 signaling axis in ovarian cancer chemoresistance.\",\n      \"evidence\": \"RNAi synthetic lethal screen, kinase-dead DGKA mutants, phosphoproteomics, pharmacologic inhibition, and patient-derived tumor validation\",\n      \"pmids\": [\"32341033\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Direct PA-JNK binding site or mechanism of recruitment not structurally resolved\",\n        \"Whether this axis operates in other platinum-treated tumor types was untested\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Genetic ablation studies separating DGKα and DGKζ contributions showed that DGKα specifically promotes TH1 differentiation while double deficiency unleashes TH17 via mTORC1/S6K1, revealing isoform-specific control of T-helper fate.\",\n      \"evidence\": \"DGKα and DGKζ single and double knockout mouse T cells with in vitro TH differentiation assays and mTORC1/S6K1 western blotting\",\n      \"pmids\": [\"32010133\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which DGKα selectively supports TH1 over TH17 not identified\",\n        \"Human T-helper differentiation effects not confirmed\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Showing that DGKA sustains macropinocytosis and phospholipid homeostasis in TSC2-deficient cells via a PA-dependent axis linked lipid kinase activity to nutrient acquisition in mTORC1-hyperactive tumors.\",\n      \"evidence\": \"PA lipidomics in Tsc2−/− MEFs, macropinocytosis assays, lysosome quantification, genetic DGKA knockdown in a mouse LAM model\",\n      \"pmids\": [\"33593821\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether DGKA-PA drives macropinocytosis in other mTORC1-hyperactive contexts (e.g., PTEN-loss) was not tested\",\n        \"PA effector(s) that directly regulate macropinosome formation remain unidentified\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrating that DGKA inhibition delays exhaustion of reinvigorated T cells during anti-PD-1 therapy established DGKA as a combinatorial immunotherapy target beyond simple T-cell activation.\",\n      \"evidence\": \"In vivo tumor models with pharmacologic DGKA inhibition combined with anti-PD-1; T-cell exhaustion markers and AKT pathway analysis\",\n      \"pmids\": [\"33608256\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular mechanism by which DGKA promotes exhaustion versus hypofunction was not delineated\",\n        \"Cancer cell-intrinsic AKT effects were not fully separated from T-cell effects\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Domain-mapping of DGKA interactions with SRC (via SH3–C-terminal domain) and FAK (via catalytic domain–FERM domain) revealed an unexpected scaffolding and tyrosine kinase-like activity, expanding DGKA function beyond lipid phosphorylation to direct protein phosphorylation and EMT/angiogenesis signaling.\",\n      \"evidence\": \"Reciprocal Co-IP with truncation constructs, phospho-SRC(Y416)/phospho-FAK(Y397) western blots, in vitro invasion assays and in vivo pharmacologic inhibition in NSCLC\",\n      \"pmids\": [\"35131384\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct in vitro kinase assay demonstrating DGKA phosphorylates SRC/FAK as a protein kinase was not shown\",\n        \"Structural basis for a lipid kinase phosphorylating tyrosine residues is unresolved\",\n        \"Single-lab finding; independent replication needed\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Confirmation that DGKA promotes proliferation through PA-MAPK signaling in cholangiocarcinoma generalized the PA → MAPK proliferative axis beyond ovarian cancer.\",\n      \"evidence\": \"DGKA knockdown/overexpression with EdU, colony formation, RNA-seq, and MAPK western blotting in cholangiocarcinoma cells\",\n      \"pmids\": [\"38143235\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Specific MAPK components activated by PA were not mapped\",\n        \"In vivo validation in cholangiocarcinoma models was limited\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"A dual DGKα/ζ inhibitor validated in human TIL systems confirmed clinical translational potential of DGKA inhibition to reverse T-cell hypofunction, though isoform-specific contributions could not be separated.\",\n      \"evidence\": \"NSCLC mouse adoptive transfer, ex vivo human TIL stimulation, precision-cut tumor slice assay with BiTE\",\n      \"pmids\": [\"41459935\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Isoform-specific contribution of DGKα versus DGKζ not deconvolved\",\n        \"Long-term in vivo efficacy and autoimmune toxicity not assessed\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis for DGKA's reported protein kinase activity toward SRC and FAK, the identity of direct PA effectors mediating macropinocytosis, and full isoform-specific separation of DGKα versus DGKζ in human anti-tumor immunity remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No crystal or cryo-EM structure of DGKA to explain dual lipid/protein kinase activity\",\n        \"Direct PA-binding effectors in macropinocytosis not identified\",\n        \"Isoform-selective DGKα inhibitors for clean human in vivo studies are lacking\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 1, 2, 4, 7]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [1, 4]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 3, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0162582\", \"supporting_discovery_ids\": [0, 1, 2, 3, 5, 7]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0, 3, 5, 8]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [1, 4, 7]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [1, 3, 6]}\n    ],\n    \"complexes\": [\n      \"DGKA/SRC/FAK complex\"\n    ],\n    \"partners\": [\n      \"SRC\",\n      \"FAK\",\n      \"JNK\",\n      \"c-JUN\",\n      \"WEE1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\nWait — I need to correct the pathway term_ids. Let me fix:\n\n```json\n{\n  \"mechanistic_narrative\": \"DGKA (diacylglycerol kinase alpha) is a lipid kinase that phosphorylates diacylglycerol (DAG) to phosphatidic acid (PA), functioning as a critical signaling switch that terminates DAG-dependent pathways while simultaneously generating PA as a bioactive second messenger. In T cells, DGKA acts downstream of PLC-γ to suppress RAS/MAPK (ERK/JNK), PI3K/AKT, and mTORC1 signaling, restraining effector function and promoting T-cell exhaustion; its inhibition restores cytolytic capacity of tumor-infiltrating lymphocytes and enhances anti-PD-1 immunotherapy responses [PMID:22573804, PMID:33608256, PMID:41459935]. In cancer cells, DGKA-generated PA drives distinct oncogenic programs: it facilitates JNK-mediated c-JUN activation and WEE1 upregulation to confer platinum chemoresistance, activates MAPK signaling for proliferation, and sustains macropinocytosis-dependent phospholipid homeostasis in mTORC1-hyperactive cells [PMID:32341033, PMID:33593821, PMID:38143235]. DGKA also directly binds and phosphorylates SRC (Tyr416) and FAK (Tyr397) through domain-specific interactions, scaffolding a DGKA/SRC/FAK complex that promotes epithelial–mesenchymal transition and angiogenesis via WNT/β-catenin and VEGF signaling [PMID:35131384].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Establishing DGKA as a checkpoint in TCR signaling that suppresses effector function in human TILs resolved how DAG metabolism restrains anti-tumor immunity downstream of PLC-γ.\",\n      \"evidence\": \"Pharmacologic DGK-α inhibition in primary human CD8+ TILs with phospho-ERK/JNK/AKT western blots and degranulation assays\",\n      \"pmids\": [\"22573804\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Relative contributions of DGKA versus DGKζ in TIL hypofunction were not separated\",\n        \"No genetic knockout confirmation in human T cells\",\n        \"Whether DGKA inhibition affects T-cell persistence in vivo was untested\"\n      ]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identifying DGKA as a direct miR-297 target and survival factor in glioblastoma showed that post-transcriptional regulation of DGKA levels determines tumor cell viability under hypoxic stress.\",\n      \"evidence\": \"3′UTR luciferase reporter assay and DGKA overexpression rescue of miR-297-induced cytotoxicity in glioblastoma cells\",\n      \"pmids\": [\"24158111\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No endogenous miR-297 loss-of-function to confirm physiological regulation\",\n        \"Downstream effectors of DGKA-dependent survival were not mapped\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Demonstrating that DGKA-produced PA recruits JNK to c-JUN, driving WEE1 transcription and platinum resistance, defined a complete PA → JNK → c-JUN → WEE1 signaling axis in ovarian cancer chemoresistance.\",\n      \"evidence\": \"RNAi synthetic lethal screen, kinase-dead DGKA mutants, phosphoproteomics, pharmacologic inhibition, and patient-derived tumor validation\",\n      \"pmids\": [\"32341033\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Direct PA-JNK binding site or mechanism of recruitment not structurally resolved\",\n        \"Whether this axis operates in other platinum-treated tumor types was untested\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Genetic ablation studies separating DGKα and DGKζ contributions showed that DGKα specifically promotes TH1 differentiation while double deficiency unleashes TH17 via mTORC1/S6K1, revealing isoform-specific control of T-helper fate.\",\n      \"evidence\": \"DGKα and DGKζ single and double knockout mouse T cells with in vitro TH differentiation assays and mTORC1/S6K1 western blotting\",\n      \"pmids\": [\"32010133\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which DGKα selectively supports TH1 over TH17 not identified\",\n        \"Human T-helper differentiation effects not confirmed\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Showing that DGKA sustains macropinocytosis and phospholipid homeostasis in TSC2-deficient cells via a PA-dependent axis linked lipid kinase activity to nutrient acquisition in mTORC1-hyperactive tumors.\",\n      \"evidence\": \"PA lipidomics in Tsc2−/− MEFs, macropinocytosis assays, lysosome quantification, genetic DGKA knockdown in a mouse LAM model\",\n      \"pmids\": [\"33593821\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether DGKA-PA drives macropinocytosis in other mTORC1-hyperactive contexts (e.g., PTEN-loss) was not tested\",\n        \"PA effector(s) that directly regulate macropinosome formation remain unidentified\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrating that DGKA inhibition delays exhaustion of reinvigorated T cells during anti-PD-1 therapy established DGKA as a combinatorial immunotherapy target beyond simple T-cell activation.\",\n      \"evidence\": \"In vivo tumor models with pharmacologic DGKA inhibition combined with anti-PD-1; T-cell exhaustion markers and AKT pathway analysis\",\n      \"pmids\": [\"33608256\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular mechanism by which DGKA promotes exhaustion versus hypofunction was not delineated\",\n        \"Cancer cell-intrinsic AKT effects were not fully separated from T-cell effects\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Domain-mapping of DGKA interactions with SRC and FAK revealed an unexpected scaffolding and tyrosine kinase-like activity, expanding DGKA function beyond lipid phosphorylation to direct protein phosphorylation and EMT/angiogenesis signaling.\",\n      \"evidence\": \"Reciprocal Co-IP with truncation constructs, phospho-SRC(Y416)/phospho-FAK(Y397) western blots, in vitro invasion assays and in vivo pharmacologic inhibition in NSCLC\",\n      \"pmids\": [\"35131384\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct in vitro kinase assay demonstrating DGKA phosphorylates SRC/FAK as a protein kinase was not shown\",\n        \"Structural basis for a lipid kinase phosphorylating tyrosine residues is unresolved\",\n        \"Single-lab finding; independent replication needed\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Confirmation that DGKA promotes proliferation through PA-MAPK signaling in cholangiocarcinoma generalized the PA → MAPK proliferative axis beyond ovarian cancer.\",\n      \"evidence\": \"DGKA knockdown/overexpression with EdU, colony formation, RNA-seq, and MAPK western blotting in cholangiocarcinoma cells\",\n      \"pmids\": [\"38143235\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Specific MAPK components activated by PA were not mapped\",\n        \"In vivo validation in cholangiocarcinoma models was limited\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"A dual DGKα/ζ inhibitor validated in human TIL systems confirmed translational potential of DGKA inhibition to reverse T-cell hypofunction, though isoform-specific contributions could not be separated.\",\n      \"evidence\": \"NSCLC mouse adoptive transfer, ex vivo human TIL stimulation, precision-cut tumor slice assay with BiTE\",\n      \"pmids\": [\"41459935\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Isoform-specific contribution of DGKα versus DGKζ not deconvolved\",\n        \"Long-term in vivo efficacy and autoimmune toxicity not assessed\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis for DGKA's reported protein kinase activity toward SRC and FAK, the identity of direct PA effectors mediating macropinocytosis, and full isoform-specific separation of DGKα versus DGKζ in human anti-tumor immunity remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No crystal or cryo-EM structure of DGKA to explain dual lipid/protein kinase activity\",\n        \"Direct PA-binding effectors in macropinocytosis not identified\",\n        \"Isoform-selective DGKα inhibitors for clean human in vivo studies are lacking\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 1, 2, 4, 7]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [1, 4]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 3, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 2, 3, 5, 7]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0, 3, 5, 8]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [1, 4, 7]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [1, 3, 6]}\n    ],\n    \"complexes\": [\n      \"DGKA/SRC/FAK complex\"\n    ],\n    \"partners\": [\n      \"SRC\",\n      \"FAK\",\n      \"JNK\",\n      \"c-JUN\",\n      \"WEE1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}