Molecule Information

General Information of the Molecule (ID: Mol04067)

• Name: Dihydroorotate dehydrogenase (DHODH) ,Homo sapiens
• Synonyms: Dihydroorotate oxidase
• Molecule Type: Protein
• Gene Name: DHODH
• Gene ID: 1723
• Location: chr16:72008588-72027664[+]
• Sequence:
  MAWRHLKKRAQDAVIILGGGGLLFASYLMATGDERFYAEHLMPTLQGLLDPESAHRLAVR
  FTSLGLLPRARFQDSDMLEVRVLGHKFRNPVGIAAGFDKHGEAVDGLYKMGFGFVEIGSV
  TPKPQEGNPRPRVFRLPEDQAVINRYGFNSHGLSVVEHRLRARQQKQAKLTEDGLPLGVN
  LGKNKTSVDAAEDYAEGVRVLGPLADYLVVNVSSPNTAGLRSLQGKAELRRLLTKVLQER
  DGLRRVHRPAVLVKIAPDLTSQDKEDIASVVKELGIDGLIVTNTTVSRPAGLQGALRSET
  GGLSGKPLRDLSTQTIREMYALTQGRVPIIGVGGVSSGQDALEKIRAGASLVQLYTALTF
  WGPPVVGKVKRELEALLKEQGFGGVTDAIGADHRR
• 3D-structure: PDB ID: 6FMD; Classification: Oxidoreductase; Method: X-ray diffraction; Resolution: 1.58 Å
• Function: Catalyzes the conversion of dihydroorotate to orotate with quinone as electron acceptor. Required for UMP biosynthesis via de novo pathway. .
• Uniprot ID: PYRD_HUMAN
• Ensembl ID: ENSG00000102967
• HGNC ID: HGNC:2867

Kingdom: Metazoa
Phylum: Chordata
Class: Mammalia
Order: Primates
Family: Hominidae
Genus: Homo
Species: Homo sapiens

Type(s) of Resistant Mechanism of This Molecule

  ADTT: Aberration of the Drug's Therapeutic Target

  MRAP: Metabolic Reprogramming via Altered Pathways

  UAPP: Unusual Activation of Pro-survival Pathway

Drug Resistance Data Categorized by Drug

Clinical Trial Drug(s) 1 drug(s) in total

BAY2402234

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Metabolic Reprogramming via Altered Pathways (MRAP)

Disease Class: Glioblastoma [ICD-11: 2A00.02] [1]

• Metabolic Type: Nucleic acid metabolism
• Sensitive Disease: Glioblastoma [ICD-11: 2A00.02]
• Sensitive Drug: BAY2402234
• Molecule Alteration: Expression; Up-regulation

Differential expression of the molecule in resistant disease

• Classification of Disease: Brain cancer [ICD-11: 2A00]
• The Specified Disease: Glioblastoma
• The Studied Tissue: Nervous tissue
• The Expression Level of Disease Section Compare with the Healthy Individual Tissue: p-value: 7.05E-05; Fold-change: 3.01E-01; Z-score: 4.81E+00
• Experimental Note: Identified from the Human Clinical Data
• In Vivo Model: HCC patients; Homo Sapiens
• Experiment for Drug Resistance: Cell viability assay
• Mechanism Description: Recently, the dihydroorotate dehydrogenase (DHODH) inhibitor BAY2402234 displayed efficacy in different brain cancer animal models

Approved Drug(s) 4 drug(s) in total

Docetaxel

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Metabolic Reprogramming via Altered Pathways (MRAP)

Disease Class: Gastric adenocarcinoma [ICD-11: 2B72.0] [2]

• Metabolic Type: Nucleic acid metabolism
• Resistant Disease: Gastric adenocarcinoma [ICD-11: 2B72.0]
• Resistant Drug: Docetaxel
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: AGS cells; Gastric; Homo sapiens (Human); CVCL_0139
• In Vitro Model: HGC27 cells; Gastric; Homo sapiens (Human); CVCL_1279
• Experiment for Molecule Alteration: qRT-PCR; Western blot analysis
• Experiment for Drug Resistance: IC50 assay
• Mechanism Description: Mechanistically, pyrimidine biosynthesis augmented Notch signaling and transcriptionally increased c-Myc expression, leading to up-regulation of critical glycolytic enzymes. Further studies revealed that pyrimidine synthesis could stabilize gamma-secretase subunit Nicastrin at post-translational N-linked glycosylation level, thereby inducing the cleavage and activation of Notch. Besides, we found that up-regulation of the key enzymes for de novo pyrimidine synthesis CAD and DHODH conferred the chemotherapeutic resistance of gastric cancer via accelerating glycolysis, and pharmacologic inhibition of pyrimidine biosynthetic pathway sensitized cancer cells to chemotherapy in vitro and in vivo.

Fluorouracil

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Metabolic Reprogramming via Altered Pathways (MRAP)

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Resistant Disease: Colorectal cancer [ICD-11: 2B91.1]
• Resistant Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Identified from the Human Clinical Data
• In Vivo Model: Patient-derived organoids; Homo Sapiens
• Experiment for Molecule Alteration: MRNA level and western blot analysis
• Experiment for Drug Resistance: Tumor volume assay
• Mechanism Description: Mechanistically, we report that intracellular lipid accumulation results in lipid peroxidation (LPO) overload, whereas mitochondrial DHODH deficiency weakens the ferroptosis defense system. The combination of these factors makes 5-FU-resistant CRC cells susceptible to ferroptosis. Moreover, mitochondrial DHODH redistribution to the cytosol increases intracellular pyrimidine pools, thereby impeding the effectiveness of 5-FU through molecular competition.

Disease Class: Gastric adenocarcinoma [ICD-11: 2B72.0] [2]

• Metabolic Type: Nucleic acid metabolism
• Resistant Disease: Gastric adenocarcinoma [ICD-11: 2B72.0]
• Resistant Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: AGS cells; Gastric; Homo sapiens (Human); CVCL_0139
• In Vitro Model: HGC27 cells; Gastric; Homo sapiens (Human); CVCL_1279
• Experiment for Molecule Alteration: qRT-PCR; Western blot analysis
• Experiment for Drug Resistance: IC50 assay
• Mechanism Description: Mechanistically, pyrimidine biosynthesis augmented Notch signaling and transcriptionally increased c-Myc expression, leading to up-regulation of critical glycolytic enzymes. Further studies revealed that pyrimidine synthesis could stabilize gamma-secretase subunit Nicastrin at post-translational N-linked glycosylation level, thereby inducing the cleavage and activation of Notch. Besides, we found that up-regulation of the key enzymes for de novo pyrimidine synthesis CAD and DHODH conferred the chemotherapeutic resistance of gastric cancer via accelerating glycolysis, and pharmacologic inhibition of pyrimidine biosynthetic pathway sensitized cancer cells to chemotherapy in vitro and in vivo.

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Resistant Disease: Colorectal cancer [ICD-11: 2B91.1]
• Resistant Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: Caco2 cells; Colon; Homo sapiens (Human); CVCL_0025
• In Vitro Model: HCT-116 cells; Colon; Homo sapiens (Human); CVCL_0291
• In Vitro Model: HCT15 cells; Colon; Homo sapiens (Human); CVCL_0292
• In Vitro Model: HCT8 5FU-R cells; Colon; Homo sapiens (Human); CVCL_2478
• In Vitro Model: HCT8 cells; Colon; Homo sapiens (Human); CVCL_2478
• In Vitro Model: HT-29 cells; Colon; Homo sapiens (Human); CVCL_0320
• In Vitro Model: LoVo 5FU-R cells; Colon; Homo sapiens (Human); CVCL_0399
• In Vitro Model: LOVO cells; Colon; Homo sapiens (Human); CVCL_0399
• In Vitro Model: SW-480 cells; Colon; Homo sapiens (Human); CVCL_0546
• In Vitro Model: T84 cells; Colon; Homo sapiens (Human); CVCL_0555
• Experiment for Molecule Alteration: MRNA level and western blot analysis
• Experiment for Drug Resistance: Cell viability assay
• Mechanism Description: Mechanistically, we report that intracellular lipid accumulation results in lipid peroxidation (LPO) overload, whereas mitochondrial DHODH deficiency weakens the ferroptosis defense system. The combination of these factors makes 5-FU-resistant CRC cells susceptible to ferroptosis. Moreover, mitochondrial DHODH redistribution to the cytosol increases intracellular pyrimidine pools, thereby impeding the effectiveness of 5-FU through molecular competition.

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Resistant Disease: Colorectal cancer [ICD-11: 2B91.1]
• Resistant Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vivo Model: Cell line-derived xenograft (CDX) models, 4-week-old male athymic BALB/c nude mice, transplanted with 5-FU-resistant organoid (PDOX5FU-R) ; cell line-derived xenograft (CDX) models, 4-week-old male athymic BALB/c nude mice, transplanted with 5-FU-resistant tumor fragment (PDX5FU-R); cell line-derived xenograft (CDX) models, 4-week-old male athymic BALB/c nude mice, transplanted with HCT8 5FU-R cells (CDXHCT8 5FU-R); cell line-derived xenograft (CDX) models, 4-week-old male athymic BALB/c nude mice, transplanted with HCT8 WT cells (CDXHCT8 WT); patient-derived xenograft (PDX) models, 4-week-old male NOG mice, transplanted with 5-FU-resistant organoid (PDOX5FU-R) ; patient-derived xenograft (PDX) models, 4-week-old male NOG mice, transplanted with 5-FU-resistant tumor fragment (PDX5FU-R); patient-derived xenograft (PDX) models, 4-week-old male NOG mice, transplanted with HCT8 5FU-R cells (CDXHCT8 5FU-R); patient-derived xenograft (PDX) models, 4-week-old male NOG mice, transplanted with HCT8 WT cells (CDXHCT8 WT); Mice
• Experiment for Molecule Alteration: MRNA level and western blot analysis
• Experiment for Drug Resistance: Tumor volume assay
• Mechanism Description: Mechanistically, we report that intracellular lipid accumulation results in lipid peroxidation (LPO) overload, whereas mitochondrial DHODH deficiency weakens the ferroptosis defense system. The combination of these factors makes 5-FU-resistant CRC cells susceptible to ferroptosis. Moreover, mitochondrial DHODH redistribution to the cytosol increases intracellular pyrimidine pools, thereby impeding the effectiveness of 5-FU through molecular competition.

Drug resistance Data Categorized by Their Corresponding Mechanisms

Metabolic Reprogramming via Altered Pathways (MRAP)

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Sensitive Disease: Colorectal cancer [ICD-11: 2B91.1]
• Sensitive Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: HT-29 cells; Colon; Homo sapiens (Human); CVCL_0320
• Experiment for Molecule Alteration: Western blot analysis
• Experiment for Drug Resistance: CCK8 assay
• Mechanism Description: Specifically, we elucidated the mechanism underlying 5-FU resistance in CRC cells, whereby the cytosolic DHODH-mediated pathway enhanced intracellular pyrimidine pools, reducing 10-FU metabolite concentrations

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Sensitive Disease: Colorectal cancer [ICD-11: 2B91.1]
• Sensitive Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: Caco2 cells; Colon; Homo sapiens (Human); CVCL_0025
• Experiment for Molecule Alteration: Western blot analysis
• Experiment for Drug Resistance: CCK8 assay
• Mechanism Description: Specifically, we elucidated the mechanism underlying 5-FU resistance in CRC cells, whereby the cytosolic DHODH-mediated pathway enhanced intracellular pyrimidine pools, reducing 11-FU metabolite concentrations

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Sensitive Disease: Colorectal cancer [ICD-11: 2B91.1]
• Sensitive Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: T84 cells; Colon; Homo sapiens (Human); CVCL_0555
• Experiment for Molecule Alteration: Western blot analysis
• Experiment for Drug Resistance: CCK8 assay
• Mechanism Description: Specifically, we elucidated the mechanism underlying 5-FU resistance in CRC cells, whereby the cytosolic DHODH-mediated pathway enhanced intracellular pyrimidine pools, reducing 12-FU metabolite concentrations

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Sensitive Disease: Colorectal cancer [ICD-11: 2B91.1]
• Sensitive Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: HCT8 cells; Colon; Homo sapiens (Human); CVCL_2478
• Experiment for Molecule Alteration: Western blot analysis
• Experiment for Drug Resistance: CCK8 assay
• Mechanism Description: Specifically, we elucidated the mechanism underlying 5-FU resistance in CRC cells, whereby the cytosolic DHODH-mediated pathway enhanced intracellular pyrimidine pools, reducing 5-FU metabolite concentrations

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Sensitive Disease: Colorectal cancer [ICD-11: 2B91.1]
• Sensitive Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: HCT15 cells; Colon; Homo sapiens (Human); CVCL_0292
• Experiment for Molecule Alteration: Western blot analysis
• Experiment for Drug Resistance: CCK8 assay
• Mechanism Description: Specifically, we elucidated the mechanism underlying 5-FU resistance in CRC cells, whereby the cytosolic DHODH-mediated pathway enhanced intracellular pyrimidine pools, reducing 6-FU metabolite concentrations

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Sensitive Disease: Colorectal cancer [ICD-11: 2B91.1]
• Sensitive Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: HCT-116 cells; Colon; Homo sapiens (Human); CVCL_0291
• Experiment for Molecule Alteration: Western blot analysis
• Experiment for Drug Resistance: CCK8 assay
• Mechanism Description: Specifically, we elucidated the mechanism underlying 5-FU resistance in CRC cells, whereby the cytosolic DHODH-mediated pathway enhanced intracellular pyrimidine pools, reducing 7-FU metabolite concentrations

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Sensitive Disease: Colorectal cancer [ICD-11: 2B91.1]
• Sensitive Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: LOVO cells; Colon; Homo sapiens (Human); CVCL_0399
• Experiment for Molecule Alteration: Western blot analysis
• Experiment for Drug Resistance: CCK8 assay
• Mechanism Description: Specifically, we elucidated the mechanism underlying 5-FU resistance in CRC cells, whereby the cytosolic DHODH-mediated pathway enhanced intracellular pyrimidine pools, reducing 8-FU metabolite concentrations

Disease Class: Colorectal cancer [ICD-11: 2B91.1] [3]

• Metabolic Type: Mitochondrial metabolism
• Sensitive Disease: Colorectal cancer [ICD-11: 2B91.1]
• Sensitive Drug: Fluorouracil
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: SW-480 cells; Colon; Homo sapiens (Human); CVCL_0546
• Experiment for Molecule Alteration: Western blot analysis
• Experiment for Drug Resistance: CCK8 assay
• Mechanism Description: Specifically, we elucidated the mechanism underlying 5-FU resistance in CRC cells, whereby the cytosolic DHODH-mediated pathway enhanced intracellular pyrimidine pools, reducing 9-FU metabolite concentrations

Leflunomide

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Unusual Activation of Pro-survival Pathway (UAPP)

Disease Class: Cholangiocarcinoma [ICD-11: 2C12.0] [4]

• Sensitive Disease: Cholangiocarcinoma [ICD-11: 2C12.0]
• Sensitive Drug: Leflunomide
• Molecule Alteration: Expression; Down-regulation
• Experimental Note: Discovered Using In-vivo Testing Model
• Cell Pathway Regulation: Mitochondrial ferroptosis regulatory signaling pathway; Regulation; N.A.
• In Vivo Model: Mouse model; Mus musculus
• Experiment for Molecule Alteration: Immunoprecipitation assay; LC-MS/MS analysis
• Experiment for Drug Resistance: Cellular ROS and lipid peroxidation level assay; LOXL3 enzymatic assay; In vitro kinase assay
• Mechanism Description: To overcome chemotherapy resistance, novel strategies sensitizing cancer cells to chemotherapy are required. Here, we screen the lysyl-oxidase (LOX) family to clarify its contribution to chemotherapy resistance in liver cancer. LOXL3 depletion significantly sensitizes liver cancer cells to Oxaliplatin by inducing ferroptosis. Chemotherapy-activated EGFR signaling drives LOXL3 to interact with TOM20, causing it to be hijacked into mitochondria, where LOXL3 lysyl-oxidase activity is reinforced by phosphorylation at S704. Metabolic adenylate kinase 2 (AK2) directly phosphorylates LOXL3-S704. Phosphorylated LOXL3-S704 targets dihydroorotate dehydrogenase (DHODH) and stabilizes it by preventing its ubiquitin-mediated proteasomal degradation. K344-deubiquitinated DHODH accumulates in mitochondria, in turn inhibiting chemotherapy-induced mitochondrial ferroptosis. CRISPR-Cas9-mediated site-mutation of mouse LOXL3-S704 to D704 causes a reduction in lipid peroxidation. Using an advanced liver cancer mouse model, we further reveal that low-dose Oxaliplatin in combination with the DHODH-inhibitor Leflunomide effectively inhibit liver cancer progression by inducing ferroptosis, with increased chemotherapy sensitivity and decreased chemotherapy toxicity.

Oxaliplatin

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Aberration of the Drug's Therapeutic Target (ADTT)

Disease Class: Cholangiocarcinoma [ICD-11: 2C12.0] [4]

• Resistant Disease: Cholangiocarcinoma [ICD-11: 2C12.0]
• Resistant Drug: Oxaliplatin
• Molecule Alteration: Missense mutation; Rv0668; p.Arg69Pro
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vivo Model: Mouse model; Mus musculus
• Experiment for Molecule Alteration: Immunoprecipitation assay; LC-MS/MS analysis
• Experiment for Drug Resistance: Cellular ROS and lipid peroxidation level assay; LOXL3 enzymatic assay; In vitro kinase assay
• Mechanism Description: To overcome chemotherapy resistance, novel strategies sensitizing cancer cells to chemotherapy are required. Here, we screen the lysyl-oxidase (LOX) family to clarify its contribution to chemotherapy resistance in liver cancer. LOXL3 depletion significantly sensitizes liver cancer cells to Oxaliplatin by inducing ferroptosis. Chemotherapy-activated EGFR signaling drives LOXL3 to interact with TOM20, causing it to be hijacked into mitochondria, where LOXL3 lysyl-oxidase activity is reinforced by phosphorylation at S704. Metabolic adenylate kinase 2 (AK2) directly phosphorylates LOXL3-S704. Phosphorylated LOXL3-S704 targets dihydroorotate dehydrogenase (DHODH) and stabilizes it by preventing its ubiquitin-mediated proteasomal degradation. K344-deubiquitinated DHODH accumulates in mitochondria, in turn inhibiting chemotherapy-induced mitochondrial ferroptosis. CRISPR-Cas9-mediated site-mutation of mouse LOXL3-S704 to D704 causes a reduction in lipid peroxidation. Using an advanced liver cancer mouse model, we further reveal that low-dose Oxaliplatin in combination with the DHODH-inhibitor Leflunomide effectively inhibit liver cancer progression by inducing ferroptosis, with increased chemotherapy sensitivity and decreased chemotherapy toxicity.

Metabolic Reprogramming via Altered Pathways (MRAP)

Disease Class: Gastric adenocarcinoma [ICD-11: 2B72.0] [2]

• Metabolic Type: Nucleic acid metabolism
• Resistant Disease: Gastric adenocarcinoma [ICD-11: 2B72.0]
• Resistant Drug: Oxaliplatin
• Molecule Alteration: Expression; Up-regulation
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: AGS cells; Gastric; Homo sapiens (Human); CVCL_0139
• In Vitro Model: HGC27 cells; Gastric; Homo sapiens (Human); CVCL_1279
• Experiment for Molecule Alteration: qRT-PCR; Western blot analysis
• Experiment for Drug Resistance: IC50 assay
• Mechanism Description: Mechanistically, pyrimidine biosynthesis augmented Notch signaling and transcriptionally increased c-Myc expression, leading to up-regulation of critical glycolytic enzymes. Further studies revealed that pyrimidine synthesis could stabilize gamma-secretase subunit Nicastrin at post-translational N-linked glycosylation level, thereby inducing the cleavage and activation of Notch. Besides, we found that up-regulation of the key enzymes for de novo pyrimidine synthesis CAD and DHODH conferred the chemotherapeutic resistance of gastric cancer via accelerating glycolysis, and pharmacologic inhibition of pyrimidine biosynthetic pathway sensitized cancer cells to chemotherapy in vitro and in vivo.

References

• Ref 1: Metabolic Plasticity of Glioblastoma Cells in Response to DHODH Inhibitor BAY2402234 Treatment. Metabolites. 2024 Jul 27;14(8):413.
• Ref 2: De novo pyrimidine synthesis fuels glycolysis and confers chemoresistance in gastric cancer. Cancer Lett. 2022 Nov 28;549:215837.
• Ref 3: Redistribution of defective mitochondria-mediated dihydroorotate dehydrogenase imparts 5-fluorouracil resistance in colorectal cancer. Redox Biol. 2024 Jul;73:103207.
• Ref 4: Lysyl oxidase-like 3 restrains mitochondrial ferroptosis to promote liver cancer chemoresistance by stabilizing dihydroorotate dehydrogenase. Nat Commun. 2023 May 30;14(1):3123.