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Cancer Science Sep 2022Lactate accumulation in the tumor microenvironment was shown to be closely related to tumor growth and immune escape, and suppression of lactate production by inhibiting...
Lactate accumulation in the tumor microenvironment was shown to be closely related to tumor growth and immune escape, and suppression of lactate production by inhibiting lactate dehydrogenase A (LDHA) has been pursued as a potential novel antitumor strategy. However, only a few potent LDHA inhibitors have been developed and most of them did not show potent antitumor effects in vivo. To this end, we designed new LDHA inhibitors and obtained a novel potent LDHA inhibitor, ML-05. ML-05 inhibited cellular lactate production and tumor cell proliferation, which was associated with inhibition of ATP production and induction of reactive oxygen species and G phase arrest. In a mouse B16F10 melanoma model, intratumoral injection of ML-05 significantly reduced lactate production, inhibited tumor growth, and released antitumor immune response of T cell subsets (Th1 and GMZB CD8 T cells) in the tumor microenvironment. Moreover, ML-05 treatment combined with programmed cell death-1 Ab or stimulator of interferon genes protein (STING) could sensitize the antitumor activity in B16F10 melanoma model. Collectively, we developed a novel potent LDHA inhibitor, ML-05, that elicited profound antitumor activity when injected locally, and was associated with the activation of antitumor immunity. In addition, ML-05 could sensitize immunotherapies, which suggests great translational value.
Topics: Animals; Antineoplastic Agents; Cell Line, Tumor; Enzyme Inhibitors; L-Lactate Dehydrogenase; Lactate Dehydrogenase 5; Lactates; Melanoma; Mice; Tumor Microenvironment
PubMed: 35722994
DOI: 10.1111/cas.15468 -
Protein Science : a Publication of the... May 2019Protein-based biological drugs and many industrial enzymes are unstable, making them prohibitively expensive. Some can be stabilized by formulation with excipients, but...
Protein-based biological drugs and many industrial enzymes are unstable, making them prohibitively expensive. Some can be stabilized by formulation with excipients, but most still require low temperature storage. In search of new, more robust excipients, we turned to the tardigrade, a microscopic animal that synthesizes cytosolic abundant heat soluble (CAHS) proteins to protect its cellular components during desiccation. We find that CAHS proteins protect the test enzymes lactate dehydrogenase and lipoprotein lipase against desiccation-, freezing-, and lyophilization-induced deactivation. Our data also show that a variety of globular and disordered protein controls, with no known link to desiccation tolerance, protect our test enzymes. Protection of lactate dehydrogenase correlates, albeit imperfectly, with the charge density of the protein additive, suggesting an approach to tune protection by modifying charge. Our results support the potential use of CAHS proteins as stabilizing excipients in formulations and suggest that other proteins may have similar potential.
Topics: Animals; Desiccation; Enzyme Stability; L-Lactate Dehydrogenase; Lipoprotein Lipase; Models, Molecular; Protein Conformation; Proteins; Tardigrada
PubMed: 30868674
DOI: 10.1002/pro.3604 -
International Journal of Molecular... Jul 2023is the most widespread cause of malaria, especially in subtropical and temperate regions such as Asia-Pacific and America. lactate dehydrogenase (PvLDH), an essential...
is the most widespread cause of malaria, especially in subtropical and temperate regions such as Asia-Pacific and America. lactate dehydrogenase (PvLDH), an essential enzyme in the glycolytic pathway, is required for the development and reproduction of the parasite. Thus, LDH from these parasites has garnered attention as a diagnostic biomarker for malaria and as a potential molecular target for developing antimalarial drugs. In this study, we prepared a transformed strain for the overexpression of PvLDH without codon optimization. We introduced this recombinant plasmid DNA prepared by insertion of the gene in the pET-21a(+) expression vector, into the Rosetta(DE3), an strain suitable for eukaryotic protein expression. The time, temperature, and inducer concentration for PvLDH expression from this Rosetta(DE3), containing the original gene, were optimized. We obtained PvLDH with a 31.0 mg/L yield and high purity (>95%) from this Rosetta(DE3) strain. The purified protein was characterized structurally and functionally. The PvLDH expressed and purified from transformed bacteria without codon optimization was successfully demonstrated to exhibit its potential tetramer structure and enzyme activity. These findings are expected to provide valuable insights for research on infectious diseases, metabolism, diagnostics, and therapeutics for malaria caused by .
Topics: Humans; Plasmodium vivax; L-Lactate Dehydrogenase; Escherichia coli; Malaria, Vivax; Malaria; Codon
PubMed: 37446261
DOI: 10.3390/ijms241311083 -
Molecules (Basel, Switzerland) Dec 2017Inhibitors of human lactate dehydrogenase (LDH5)-the enzyme responsible for the conversion of pyruvate to lactate coupled with oxidation of NADH to NAD⁺-are promising... (Review)
Review
Inhibitors of human lactate dehydrogenase (LDH5)-the enzyme responsible for the conversion of pyruvate to lactate coupled with oxidation of NADH to NAD⁺-are promising therapeutic agents against cancer because this enzyme is generally found to be overexpressed in most invasive cancer cells and is linked to their vitality especially under hypoxic conditions. Consequently, significant efforts have been made for the identification of small-molecule LDH5 inhibitors displaying high inhibitory potencies. X-ray structure of LDH5 complexes as well as molecular modeling studies contribute to identify and explain the main binding modes of LDH5 inhibitors reported in literature. The purpose of this review is to analyze the main three-dimensional interactions between some of the most potent inhibitors and LDH5, in order to provide useful suggestions for the design of new derivatives.
Topics: Antineoplastic Agents; Binding Sites; Crystallography, X-Ray; Enzyme Inhibitors; Gene Expression; Humans; Hydrogen Bonding; Hydrophobic and Hydrophilic Interactions; L-Lactate Dehydrogenase; Molecular Docking Simulation; NAD; Neoplasm Proteins; Neoplasms; Protein Binding; Protein Conformation; Protein Interaction Domains and Motifs
PubMed: 29236080
DOI: 10.3390/molecules22122217 -
Cellular & Molecular Biology Letters 2020One of the hallmarks of cancer cells is increased energy requirements associated with the higher rate of cellular proliferative activity. Metabolic changes in rapidly... (Review)
Review
One of the hallmarks of cancer cells is increased energy requirements associated with the higher rate of cellular proliferative activity. Metabolic changes in rapidly dividing cancer cells are closely associated with increased uptake of glucose and abnormal activity of lactate dehydrogenase (LDH), which regulates the processing of glucose to lactic acid. As serum LDH levels were found to be commonly increased in cancer patients and correlated with poor clinical outcome and resistance to therapy, the determination of LDH has become a standard supportive tool in diagnosing cancers or monitoring the effects of cancer treatment. The aim of this review is to summarize the current knowledge about methods and the practical utility for measuring both the total LDH and LDH isoenzymatic activities in the diagnosis, prognosis and prediction of cancer diseases.
Topics: Biomarkers, Tumor; Colorimetry; Humans; Isoenzymes; L-Lactate Dehydrogenase; Neoplasms; Prognosis
PubMed: 32528540
DOI: 10.1186/s11658-020-00228-7 -
Scientific Reports Jul 2017Lactate dehydrogenase A (LDHA) has been reported to be involved in the initiation and progression of tumors. However, the potential role of LDHA in pituitary adenoma...
Lactate dehydrogenase A (LDHA) has been reported to be involved in the initiation and progression of tumors. However, the potential role of LDHA in pituitary adenoma (PA) remains unknown. In this study, we showed that the expression levels of LDHA mRNA and protein were significantly elevated in invasive PA samples, and positively correlated with higher Ki-67 index. Overexpression of LDHA in a PA cell line (GH3) promoted glucose uptake through the upregulation of glucose transporter-1 (Glut1), lactate secretion and induced cellular invasion by upregulation of matrix metalloproteinase2 (MMP2). LDHA also promoted GH3 cell proliferation through induction of cell cycle progression via activation of the Akt-GSK-3β-cyclinD1 pathway. Accordingly, oxamate-induced inhibition of LDHA suppressed glucose uptake, lactate secretion, invasion and proliferation in GH3 cells via down regulation of Glut1 and MMP2 expression and inhibition of the Akt-GSK-3β-cyclinD1 pathway. Moreover, oxamate induced GH3 cell apoptosis by increasing mitochondrial reactive oxygen species (ROS) generation. In vivo, LDHA overexpression promoted tumor growth, and oxamate delayed tumor growth. In primary PA cell cultures, oxamate also effectively suppressed invasion and proliferation. Our data indicate that LDHA is involved in promoting the progression of PA, and oxamate might be a promising therapeutic agent for the treatment of PA.
Topics: Adenoma; Animals; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Glucose; Humans; Isoenzymes; L-Lactate Dehydrogenase; Lactate Dehydrogenase 5; Mice; Neoplasm Invasiveness; Neoplasm Transplantation; Pituitary Neoplasms; Signal Transduction; Up-Regulation
PubMed: 28680051
DOI: 10.1038/s41598-017-04366-5 -
PloS One 2017In the asexual stages, Toxoplasma gondii stage converts between acute phase rapidly replicating tachyzoites and chronic phase slowly dividing bradyzoites....
In the asexual stages, Toxoplasma gondii stage converts between acute phase rapidly replicating tachyzoites and chronic phase slowly dividing bradyzoites. Correspondingly, T. gondii differentially expresses two distinct genes and isoforms of the lactate dehydrogenase enzyme, expressing LDH1 exclusively in the tachyzoite stage and LDH2 preferentially in the bradyzoite stage. LDH catalyzes the interconversion of pyruvate and lactate in anaerobic growth conditions and is utilized for energy supply, however, the precise role of LDH1 and LDH2 in parasite biology in the asexual stages is still unclear. Here, we investigated the biological role of LDH1 and LDH2 in the asexual stages, and the vaccine strain potential of deletion mutants lacking LDH1, LDH2, or both genes (Δldh1, Δldh2 and Δldh1/2). Deletion of LDH1 reduced acute parasite virulence, impaired bradyzoite differentiation in vitro, and markedly reduced chronic stage cyst burdens in vivo. In contrast, deletion of LDH2 impaired chronic stage cyst burdens without affecting virulence or bradyzoite differentiation. Deletion of both LDH1 and LDH2 induced a more severe defect in chronic stage cyst burdens. These LDH mutant phenotypes were not associated with any growth defect. Vaccination of mice with a low dose of mutants deleted for LDH elicited effective protective immunity to lethal challenge infection, demonstrating the vaccine potential of LDH deletion mutants. These results suggest that lactate dehydrogenase in T. gondii controls virulence, bradyzoite differentiation, and chronic infection and reveals the potential of LDH mutants as vaccine strains.
Topics: Amino Acid Sequence; Animals; Cells, Cultured; Escherichia coli; Female; Gene Knockout Techniques; Humans; Isoenzymes; L-Lactate Dehydrogenase; Mice, Inbred BALB C; Mutation; Random Allocation; Recombinant Proteins; Toxoplasma; Toxoplasmosis; Vaccination; Virulence
PubMed: 28323833
DOI: 10.1371/journal.pone.0173745 -
International Journal of Biological... Dec 2020The binding between the enzyme lactate dehydrogenase (LDH) and ferrihydrite nanoparticles (Fh-NPs) was investigated by means of small-angle neutron scattering (SANS),...
The binding between the enzyme lactate dehydrogenase (LDH) and ferrihydrite nanoparticles (Fh-NPs) was investigated by means of small-angle neutron scattering (SANS), Fourier-transform infrared (FTIR) spectroscopy, fluorescence and Förster resonance energy transfer (FRET) and molecular docking. Fh-NPs - LDH compounds of dimensions under 100 nm are formed. The conformational changes and the mechanism of interaction between LDH and Fh-NPs simple and doped with Cu and Co, and the effect of these NPs on the thermal denaturation of LDH were monitored. The quenching mechanism is static, the binding occurring with moderate affinity, being mainly driven by hydrogen bonding and van der Waals forces. FRET occurs at a minimal distance of 2.55 nm. Thermal denaturation of LDH in the presence of simple and doped Fh-NPs shows that the thermodynamic parameters of protein unfolding are significantly changed with temperature. The denaturation temperature of LDH shifts to higher values in the presence of all Fh-NPs, than in the case of simple LDH. The docking approach estimates the energy corresponding to the best fit of the ferrihydrite in the LDH binding site near Trp. These results have direct implications on the uses of the complex of LDH with Fh-NPs in various biochemical, biological, or clinical applications.
Topics: Algorithms; Chemical Phenomena; Drug Discovery; Ferric Compounds; L-Lactate Dehydrogenase; Models, Theoretical; Molecular Docking Simulation; Molecular Dynamics Simulation; Molecular Structure; Nanoparticles; Protein Binding; Spectrum Analysis; Structure-Activity Relationship; Thermodynamics
PubMed: 32890566
DOI: 10.1016/j.ijbiomac.2020.08.242 -
Future Medicinal Chemistry Mar 2020Dysregulated metabolism is one of the hallmarks of cancer. Under normal physiological conditions, ATP is primarily generated by oxidative phosphorylation. Cancers...
Dysregulated metabolism is one of the hallmarks of cancer. Under normal physiological conditions, ATP is primarily generated by oxidative phosphorylation. Cancers commonly undergo a dramatic shift toward glycolysis, despite the presence of oxygen. This phenomenon is known as the Warburg effect, and requires the activity of LDHA. LDHA converts pyruvate to lactate in the final step of glycolysis and is often upregulated in cancer. LDHA inhibitors present a promising therapeutic option, as LDHA blockade leads to apoptosis in cancer cells. Despite this, existing LDHA inhibitors have shown limited clinical efficacy. Here, we review recent progress in LDHA structure, function and regulation as well as strategies to target this critical enzyme.
Topics: Apoptosis; Enzyme Inhibitors; Humans; L-Lactate Dehydrogenase; Neoplasms; Protein Conformation
PubMed: 32064930
DOI: 10.4155/fmc-2019-0287 -
Bioorganic & Medicinal Chemistry Letters Jun 2021Lactate dehydrogenase (LDH) is a critical enzyme in the glycolytic metabolism pathway that is used by many tumor cells. Inhibitors of LDH may be expected to inhibit the...
Lactate dehydrogenase (LDH) is a critical enzyme in the glycolytic metabolism pathway that is used by many tumor cells. Inhibitors of LDH may be expected to inhibit the metabolic processes in cancer cells and thus selectively delay or inhibit growth in transformed versus normal cells. We have previously disclosed a pyrazole-based series of potent LDH inhibitors with long residence times on the enzyme. Here, we report the elaboration of a new subseries of LDH inhibitors based on those leads. These new compounds potently inhibit both LDHA and LDHB enzymes, and inhibit lactate production in cancer cell lines.
Topics: Aniline Compounds; Antineoplastic Agents; Cell Line, Tumor; Drug Design; Ethers; Humans; L-Lactate Dehydrogenase
PubMed: 33771585
DOI: 10.1016/j.bmcl.2021.127974