Monocarboxylate Transporter Isoform Research Articles

Five Weeks of Sprint Interval Training Improve Muscle Glycolytic Content and Activity But Not Time to Task Failure in Severe-Intensity Exercise.

This study examined the impact of a 5-wk sprint interval training (SIT) intervention on time to task failure (TTF) during severe-intensity constant work rate (CWR) exercise, as well as in glycolytic enzymatic content and activity, and glycogen content. Fourteen active males were randomized into either a SIT group ( n = 8) composed of 15 SIT sessions over 5 wk, or a control group ( n = 6). At pretraining period, participants performed i) ramp incremental test to measure the cardiorespiratory function; ii) CWR cycling TTF at 150% of the power output (PO) at the respiratory compensation point (RCP-PO) with muscle biopsies at rest and immediately following task failure. After 5 wk, the same evaluations were repeated (i.e., exercise intensities matched to current training status), and an additional cycling CWR matched to pretraining 150% RCP-PO was performed only for TTF evaluation. The content and enzymatic activity of glycogen phosphorylase (GPhos), hexokinase (HK), phosphofructokinase (PFK), and lactate dehydrogenase (LDH), as well as the glycogen content, were analyzed. Content of monocarboxylate transporter isoform 4 (MCT4) and muscle buffering capacity were also measured. Despite improvements in total work performed at CWR posttraining, no differences were observed for TTF. The GPhos, HK, PFK, and LDH content and activity, and glycogen content also improved after training only in the SIT group. Furthermore, the MCT4 concentrations and muscle buffering capacity were also improved only for the SIT group. However, no difference in glycogen depletion was observed between groups and time. Five weeks of SIT improved the glycolytic pathway parameters and total work performed; however, glycogen depletion was not altered during CWR severe-intensity exercise, and TTF remained similar.

Disposition of glycolic acid into the embryo following oral administration of ethylene glycol during placentation in the rat and rabbit.

In order to evaluate the role of the placenta in the etiology of ethylene glycol (EG) developmental toxicity, the distribution of EG and its main metabolites, glycolic acid (GA) and oxalic acid (OX), into the conceptus was determined at the beginning and completion of placentation in the rat and rabbit. Two groups (n = 28) of timed-pregnant Wistar rats were administered EG (1000 mg/kg bw/day, oral gavage) from gestation day (GD) 6 to either GD 11 or GD 16; similarly, two groups (n = 28) of timed-pregnant New Zealand White rabbits were administered EG from GD 6 to either GD 10 or GD 19. Four animals from each group were sacrificed at 1, 3, 6, 9, 12, 18, or 24 h after the final administration, and maternal blood, extraembryonic fluid, and embryonic tissue were removed for analysis of EG, GA, and OX. The three analytes were predominantly cleared from all compartments in both species within 24 h. Neither EG nor OX preferentially accumulated into the conceptus compartments, compared with the maternal blood, in either species. Critically, GA was preferentially accumulated from the maternal blood only into the rat embryo at GD 11, but not at GD 16 and not into the rabbit embryo at either GD 10 or GD 19. The accumulation of GA into the rat embryo, and its decline over the course of placentation, is discussed in relation to the expression of monocarboxylate transporter isoforms across the syncytiotrophoblast.

Ensiled diet improved the growth performance of Tibetan sheep by regulating the rumen microbial community and rumen epithelial morphology.

The aim of this study was to investigate the effects of ensiled agricultural byproducts from Qinghai-Tibet plateau on growth performance, rumen microbiota, ruminal epithelium morphology, and nutrient transport-related gene expression in Tibetan sheep. Fourteen male Tibetan sheep were randomly assigned to one of two diets: an untreated diet (without silage inoculum, CON, n = 7) or an ensiled diet (with silage inoculum, ESD, n = 7). The total experimental period lasted for 84 d, including early 14 d as adaption period and remaining 70 d for data collection. The ESD increased average daily gain (P = 0.046), dry matter intake (P < 0.001), ammonia nitrogen (P = 0.045), microbial crude protein (P = 0.034), and total volatile fatty acids concentration (P < 0.001), and decreased ruminal pH value (P = 0.014). The proportion of propionate (P = 0.006) and the copy numbers of bacteria (P = 0.01) and protozoa (P = 0.002) were higher, while the proportion of acetate (P = 0.028) was lower in the sheep fed ESD compared to CON. Pyrosequencing of the 16S ribosomal RNA gene revealed that ESD increased the relative abundance of Firmicutes, Ruminococcus, Lachnospiraceae_AC2044_group, Lachnospiraceae_XPB1014_group, and Christensenellaceae_R-7_group in the rumen (P < 0.05), while decreased the relative abundance of Bacteroidota, Prevotellaceae_UCG-003, and Veillonellaceae_UCG-001 (P < 0.05). Analyses with PICRUSt2 and STAMP indicated that the propionate metabolism pathway was enriched in the sheep fed ESD (P = 0.026). The ESD increased the rumen papillae height (P = 0.012), density (P = 0.036), and surface area (P = 0.001), and improved the thickness of the total epithelia (P = 0.018), stratum corneum (P = 0.040), stratum granulosum (P = 0.042), and stratum spinosum and basale (P = 0.004). The relative mRNA expression of cyclin-dependent Kinase 2, CyclinA2, CyclinD2, zonula occludens-1, Occludin, monocarboxylate transporter isoform 1 (MCT1), MCT4, sodium/potassium pump, and sodium/hydrogen antiporter 3 were higher in the rumen epithelial of sheep fed ESD than CON (P < 0.05). Conversely, the relative mRNA expressions of Caspase 3 and B-cell lymphoma-2 were lower in the sheep fed ESD than CON (P < 0.05). In conclusion, compared with an untreated diet, feeding an ensiled diet altered the rumen microbial community, enhanced nutrient transport through rumen epithelium, and improved the growth performance of Tibetan sheep.

Monocarboxylate Transporters: Role and Regulation in Corneal Diabetes.

Diabetes mellitus (DM) is a group of metabolic diseases that is known to cause structural and functional ocular complications. In the human cornea, DM-related complications affect the epithelium, stroma, and nerves. Monocarboxylate transporters (MCTs) are a family of proton-linked plasma membrane transporters that carry monocarboxylates across plasma membranes. In the context of corneal health and disease, their role, presence, and function are largely undetermined and solely focused on the most common MCT isoforms, 1 through 4. In this study, we investigated the regulation of MCT1, 2, 4, 5, 8, and 10, in corneal DM, using established 3D self-assembled extracellular matrix (ECM) in vitro models. Primary stromal corneal fibroblasts were isolated from healthy (HCFs), type I (T1DMs), and type II (T2DMs) DM donors. Monoculture 3D constructs were created by stimulating stromal cells on transwells with stable vitamin C for two or four weeks. Coculture 3D constructs were created by adding SH-SY5Y neurons at two different densities, 12 k and 500 k, on top of the monocultures. Our data showed significant upregulation of MCT1 at 4 weeks for HCF, T1DM, and T2DM monocultures, as well as the 500 k nerve cocultures. MCT8 was significantly upregulated in HCF and T1DM monocultures and all of the 500 k nerve cocultures. Further, MCT10 was only expressed at 4 weeks for all cocultures and was limited to HCFs and T1DMs in monocultures. Immunofluorescence analysis showed cytoplasmic MCT expression for all cell types and significant downregulation of both MCT2 and MCT4 in HCFs, when compared to T1DMs and T2DMs. Herein, we reveal the existence and modulation of MCTs in the human diabetic cornea in vitro. Changes appeared dependent on neuronal density, suggesting that MCTs are very likely critical to the neuronal defects observed in diabetic keratopathy/neuropathy. Further studies are warranted in order to fully delineate the role of MCTs in corneal diabetes.

Involvement of SLC16A1/MCT1 and SLC16A3/MCT4 in l-lactate transport in the hepatocellular carcinoma cell line.

Fourteen isoforms of the monocarboxylate transporter (MCT) have been reported. Among the MCT isoforms, MCT1, MCT2, and MCT4 play a role in l-lactate/proton cotransport and are involved in the balance of intracellular energy and pH. Therefore, MCT1, MCT2, and MCT4 are associated with energy metabolism processes in normal and pathological cells. In the present study, we evaluated the expression of MCT1, MCT2, and MCT4 and the contribution of these three MCT isoforms to l-lactate uptake in hepatocellular carcinoma (HCC) cells. In HepG2 and Huh-7 cells, l-lactate transport was pH-dependent, which is characteristic of MCT1, MCT2, and MCT4. Furthermore, l-lactate uptake was selectively inhibited by MCT1 and MCT4 inhibitors in HepG2 and Huh-7 cells. Kinetic analysis of HepG2 cells demonstrated that l-lactate uptake was biphasic. Although the knockdown of MCT1 and MCT4 in the HepG2 cells decreased the uptake of l-lactate, the knockdown of MCT2 had no effect on the uptake of l-lactate. Consequently, we concluded that both MCT1 and MCT4 were involved in the transport of l-lactate in HepG2 and Huh-7 cells at pH 6.0. In contrast, PXB-cells, freshly isolated hepatocytes from humanized mouse livers, showed lower MCT4 expression and l-lactate uptake at pH 6.0 compared to that in HCC cell lines. In conclusion, MCT4, which contributes to l-lactate transport in HCC cells, is significantly different in HCC compared to normal hepatocytes, and has potential as a target for HCC treatment.

Differential response of hepatocellular carcinoma glycolytic metabolism and oxidative stress markers after exposure to human amniotic membrane proteins.

The human Amniotic Membrane (hAM) has been studied as a potential therapeutic option in cancer, namely in hepatocellular carcinoma. Previously, our research group evaluated the effect of human Amniotic Membrane Protein Extracts (hAMPE) in cancer therapy, demonstrating that hAMPE inhibit the metabolic activity of human hepatocellular carcinoma cell lines: Hep3B2.1-7, HepG2 and Huh7. Therefore, and considering the close relationship between metabolic activity and oxidative stress, the aim of this study was to evaluate the effect of hAMPE treatment in glucose metabolism and its role in oxidative stress of hepatocellular carcinoma. Glucose uptake and lactate production was assessed by 1H-NMR, and the expression of several mediators of the glycolytic pathway was evaluated by Western blot or fluorescence. Total antioxidant capacity (TAC) and biomarkers of oxidative stress effects in proteins were detected. Our results showed that hAMPE treatment increased glucose consumption on Hep3B2.1-7, HepG2, and Huh7 through the increase of GLUT1 in Hep3B2.1-7 and Huh7, and GLUT3 in HepG2 cells. It was observed an increased expression of 6-phosphofrutokinase (PFK-1L) in all cell lines though glucose was not converted to lactate on HepG2 and Huh7 cells, suggesting that hAMPE treatment may counteract the Warburg effect observed in carcinogenesis. In Hep3B2.1-7, hAMPE treatment induced an increase in expression of lactate dehydrogenase (LDH) and monocarboxylate transporter isoform 4 (MCT4). We further detected that hAMPE enhances the TAC of culture media after 2 and 8h. This was followed by a degree of protection against proteins nitration and carbonylation. Overall, this work highlights the potential usefulness of hAMPE as anticancer therapy through the modulation of the glycolytic and oxidative profile in human hepatocellular carcinoma.

PD43-03 MCT INHIBITION SYNERGIZES WITH TYROSINE KINASE AND MTOR INHIBITOR TREATMENT IN RENAL CELL CARCINOMA

You have accessJournal of UrologyCME1 May 2022PD43-03 MCT INHIBITION SYNERGIZES WITH TYROSINE KINASE AND MTOR INHIBITOR TREATMENT IN RENAL CELL CARCINOMA Bhaghyasree Jambunathan, Abdo Dergham, Sayani Bhattacharjee, Rebecca Wynn, Puneet Sindhwani, Firas Petros, and Nagalakshmi Nadiminty Bhaghyasree JambunathanBhaghyasree Jambunathan More articles by this author , Abdo DerghamAbdo Dergham More articles by this author , Sayani BhattacharjeeSayani Bhattacharjee More articles by this author , Rebecca WynnRebecca Wynn More articles by this author , Puneet SindhwaniPuneet Sindhwani More articles by this author , Firas PetrosFiras Petros More articles by this author , and Nagalakshmi NadimintyNagalakshmi Nadiminty More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002604.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: RCC has been recognized in recent years as a uniquely metabolic disease. All histologic types of RCC exhibit increase in aerobic glycolysis with disease progression. The TCGA-RCC study showed that high grade, high stage, and low survival are associated with a shift to aerobic glycolysis and a decrease in oxidative phosphorylation. Approaches targeting glucose metabolism in RCC such as GLUT1 inhibitors, hexokinase inhibitors, GAPDH inhibitor, and PKM2 inhibitor showed limited clinical benefit. Monocarboxylate transporters (MCTs) play a major role in the export/import of the glycolytic end-product lactate. Different MCT isoforms are differentially expressed in RCC tissues with an increase in the expression of MCT1 and MCT4 in clear-cell RCC and papillary RCC. Here we hypothesized that MCT inhibition may be an attractive therapeutic strategy against RCC cells. METHODS: We analyzed the expression levels of MCTs in RCC tissues using public datasets from Oncomine. We treated normal renal epithelial and renal cell carcinoma cells with varying concentrations of MCT inhibitors AR-C155858, AZD3965, or syrosingopine either singly or in combination with sunitinib or everolimus and assessed cell survival, cell viability, proliferation, clonogenic ability, survival in 3-D models, and glycolytic activity. We treated xenografts of ccRCC cells in mice with MCT inhibitors, sunitinib, or everolimus singly or in combination and assessed tumor growth. RESULTS: We found that the suppression of MCT activity using the MCT antagonists AR-C155858, AZD3965, or syrosingopine not only diminished the proliferation, survival, and clonogenic ability of RCC cells, but also potentiated the response of RCC cells to treatment with sunitinib or everolimus. MCT inhibitors inhibited the metabolic adaptation of ccRCC cells when treated with sunitinib or everolimus. Tumor growth of ccRCC xenografts was suppressed significantly when treated with a combination of MCT inhibitors with sunitinib or everolimus. These findings imply that MCT inhibition may be an effective strategy to develop combinatorial therapies in renal cell carcinoma. CONCLUSIONS: MCT inhibition may improve the anti-tumor effects of tyrosine kinase or mTOR inhibitors in renal cell carcinoma. Source of Funding: DoD-PCRP PC190332 (W81XWH2010794), University of Toledo College of Medicine and Life Sciences, NIH/NCI 1R21CA202404, NIH/NCI 1R03CA198696 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e702 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Bhaghyasree Jambunathan More articles by this author Abdo Dergham More articles by this author Sayani Bhattacharjee More articles by this author Rebecca Wynn More articles by this author Puneet Sindhwani More articles by this author Firas Petros More articles by this author Nagalakshmi Nadiminty More articles by this author Expand All Advertisement PDF DownloadLoading ...

Effects of dietary oil sources (sunflower and fish) on fermentation characteristics, epithelial gene expression and microbial community in the rumen of lambs fed a high-concentrate diet

ABSTRACT The feeding of high-concentrate diets commonly results in lowered pH and ruminal dysbiosis which cause shifts in uptake dynamics of short-chain fatty acids (SCFA) and altered epithelial function. Therefore, the current study evaluated the effect of dietary polyunsaturated fatty acids (PUFA) on ruminal fermentation products, gene expression in the ruminal epithelium and the associated changes in ruminal microorganisms in lambs fed a high-concentrate diet. Twenty-six Afshari lambs adapted to a high-concentrate diet during a completely randomised design were fed with a basal diet supplemented with 100 g oil supplement (OS; 60 g sunflower oil and 40 g fish oil) for 10 (OS10), 20 (OS20) and 30 (OS30) d, respectively (n = 6). Lambs with no oil supplementation (OS0, n = 8) were considered as control and slaughtered at d 0 of the experiment, and the remaining lambs were slaughtered at 10, 20 and 30 d on feed. After slaughter, ruminal digesta was collected for evaluating fermentation and microbial community. Ruminal papillae were taken for assessment of epithelial gene expression. Compared with OS0 lambs, supplemental PUFA in OS30 lambs tended to decrease total SCFA concentration with decreased acetic and increased propionic acid concentrations. Acetate:propionate ratios were decreased and ruminal pH was increased in OS20 and OS30 lambs compared to OS0. All groups with included OS had decreased concentrations of iso-valeric and valeric acids compared to OS0. Relative mRNA abundance of monocarboxylate transporter isoforms 1 and 4, insulin-like growth factor binding protein 3, sterol regulatory element-binding proteins 1 and 2 decreased with increasing OS duration. The relative abundance of 3-hydroxy-3-methylglutaryl-CoA synthase 1 mRNA transcript was higher for OS10 and OS20 lambs relative to OS0 lambs. OS20 and OS30 showed a decrease of lipopolysaccharide binding protein mRNA expression compared with OS0. Feeding supplemental PUFA decreased Ciliate protozoa and increased Butyrivibrio fibrisolvens in OS20 and OS30 lambs, whereas Megasphaera elsdenii was increased in OS30 lambs. In conclusion, combined supplementation of sunflower and fish oil to a high-concentrate diet affects the ruminal microbial community with prominent decreases in ruminal ciliate protozoa and increases in B. fibrisolvens and M. elsdenii. These results lead to a more stabilised ruminal pH and a fermentation shift towards more propionate generation. Consideration of nutrients digestion will help to fully understand the benefits of feeding PUFA with a high-concentrate diet.

Effects of weaning on regulators of volatile fatty acid absorption and intracellular pH in Holstein calves

This study examined the effects of changes in rumen fermentation during the weaning transition on abundance of transporters involved in volatile fatty acid (VFA) absorption or intracellular pH homeostasis. Holstein bull calves (n = 27) were assigned to 1 of 3 treatment groups in a randomized, complete block design: 2 preweaning groups [animals fed milk only (PRE-M) or milk, calf starter, and hay (PRE-S)] and 1 postweaning group (animals fed milk, starter, and hay with a 2-wk weaning transition; POST-S). Calves were euthanized at 42 d of age (PRE-M and PRE-S) or at 63 d of age (POST-S), and rumen epithelium and rumen fluid samples were collected. Rumen fluid was analyzed for VFA concentration, and rumen epithelium was analyzed for the abundance of VFA transporter monocarboxylate transporter isoform 1 (MCT1) and the intracellular pH regulators sodium bicarbonate co-transporter 1 (NBC1) and sodium-proton exchanger 3 (NHE3) protein. Preweaning, total VFA concentrations tended to increase and NBC1 abundance increased with starter intake. Between pre- and postweaning, total VFA concentrations increased but NHE3 protein abundance decreased. In calves, rumen epithelial development during the weaning transition appears to show more pronounced changes in intracellular pH homeostasis than in VFA transport capacity.

Abstract 2330: MCT inhibition as a therapeutic strategy to target Enzalutamide-resistant prostate cancer

Abstract Sayani Bhattacharjee, Jonathan Doan, Rebecca Wynn, Nagalakshmi Nadiminty Enzalutamide is a second-generation anti-androgen used for the treatment of prostate cancer. However, almost all patients develop resistance to this drug, thereby creating an urgent need of identifying new approaches that can overcome said resistance. Using parental and Enzalutamide-resistant C4-2B and 22RV1 cells, we targeted cellular metabolism to beat therapeutic resistance. Prostate cancers are more heterogeneous in their use of energy sources compared with other solid tumors and the most common metabolic end product is lactate, which is toxic to cancer cells upon accumulation. Hence, cancer cells upregulate the expression of monocarboxylate transporters (MCTs) that play a pivotal role in lactate efflux. Different MCT isoforms are differentially expressed across prostate cancer progression and presumably according to cellular demands at different stages. Gradual progression from benign prostate, to prostatic intraepithelial neoplasia, to in-situ carcinoma showed an increase in the expression of MCT2 and MCT4, with MCT4 being expressed in invasive prostate cancers. Based on our preliminary data that the expression of MCTs is higher in prostate cancer cells resistant to enzalutamide, we hypothesized that MCT inhibition may be an attractive therapeutic strategy against enzalutamide-resistant prostate cancer cells. Using cell survival, cell proliferation and clonogenic-ability assays, we found that the suppression of MCT activity using the MCT1/2 antagonists AR-C155858 or AZD3965 and the MCT 4 antagonist Syrosingopine not only diminishes the proliferation and survival of prostate cancer cells, but also re-sensitizes resistant cells to treatment with enzalutamide. We also found that treatment with AR-C155858 or AZD3965 either singly, or in combination with Enzalutamide suppressed prostate cancer cell xenograft growth in SCID mice bearing subcutaneously injected parental or Enzalutamide-resistant C4-2B cells. These findings imply that the MCTs have an intrinsic role in the acquisition of resistance to therapy. Citation Format: MCT inhibition as a therapeutic strategy to target Enzalutamide-resistant prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2330.

Cysteine 159 delineates a hinge region of the alternating access monocarboxylate transporter1 and is targeted by cysteine-modifying inhibitors.

Monocarboxylate transporter isoforms 1-4, MCT, of the solute carrier SLC16A family facilitate proton-coupled transport of l-lactate. Growth of tumors that exhibit the Warburg effect, that is, high rates of anaerobic glycolysis despite availability of oxygen, relies on swift l-lactate export, whereas oxygenic cancer cells import circulating l-lactate as a fuel. Currently, MCTs are viewed as promising anticancer targets. Small-molecule inhibitors have been found, and, recently, high-resolution protein structures have been obtained. Key questions, however, regarding the exact binding sites of cysteine-modifying inhibitors and the substrate translocation cycle lack a conclusive experimental basis. Here, we report Cys159 of the ubiquitous human MCT1 to reside in a critical hinge region of the alternating access-type transporter. We identified Cys159 as the binding site of the organomercurial pCMBS. The inhibitory effect of pCMBS was proposed to be indirect via modification of the chaperone basigin. We provide evidence that pCMBS locks MCT1 in its outward open conformation in a wedge-like fashion. We corroborated this finding using smaller cysteine-modifying reagents that size-dependently inhibited l-lactate transport. The smallest modifiers targeted additional cysteines as shown by a C159S mutant. We found a Cys399/Cys400 pair to constitute the second hinge of the transporter that tolerated only individual replacement by serine. The hinge cysteines, in particular the selectively addressable Cys159, provide natural anchors for placing probes into MCTs to report, for instance, on the electrostatics or hydration upon binding of the transported l-lactate substrate and the proton cosubstrate.

Modeling hyperpolarized lactate signal dynamics in cells, patient-derived tissue slice cultures and murine models.

Determining the aggressiveness of renal cell carcinoma (RCC) noninvasively is a critical part of the diagnostic workup for treating this disease that kills more than 15,000 people annually in the USA. Recently, we have shown that not only the amount of lactate produced, as a consequence of the Warburg effect, but also its efflux out of the cell, is a critical marker of RCC aggressiveness and differentiating RCCs from benign renal tumors. Enzymatic conversions can now be measured in situ with hyperpolarized (HP) 13 C magnetic resonance (MR) on a sub-minute time scale. Using RCC models, we have shown that this technology can interrogate in real time both lactate production and compartmentalization, which are associated with tumor aggressiveness. The dynamic HP MR data have enabled us to robustly characterize parameters that have been elusive to measure directly in intact living cells and murine tumors thus far. Specifically, we were able to measure the same intracellular lactate longitudinal relaxation time in three RCC cell lines of 16.42 s, and lactate efflux rate ranging from 0.14 to 0.8 s-1 in the least to the most aggressive RCC cell lines and correlate it to monocarboxylate transporter isoform 4 expression. We also analyzed dynamic HP lactate and pyruvate data from orthotopic murine RCC tumors using a simplified one-compartment model, and showed comparable apparent pyruvate to lactate conversion rate (kPL ) values with those measured in vitro. This kinetic modeling was then extended to characterize the lactate dynamics in patient-derived living RCC tissue slices; and even without direct measurement of the extracellular lactate signal the efflux parameter was still assessed and was distinct between the benign renal tumors and RCCs. Across all these preclinical models, the rate parameters of kPL and lactate efflux correlated to cancer aggressiveness, demonstrating the validity of our modeling approach for noninvasive assessment of RCC aggressiveness.

Monocarboxylate Transporters (SLC16): Function, Regulation, and Role in Health and Disease.

The solute carrier family 16 (SLC16) is comprised of 14 members of the monocarboxylate transporter (MCT) family that play an essential role in the transport of important cell nutrients and for cellular metabolism and pH regulation. MCTs 1-4 have been extensively studied and are involved in the proton-dependent transport of L-lactate, pyruvate, short-chain fatty acids, and monocarboxylate drugs in a wide variety of tissues. MCTs 1 and 4 are overexpressed in a number of cancers, and current investigations have focused on transporter inhibition as a novel therapeutic strategy in cancers. MCT1 has also been used in strategies aimed at enhancing drug absorption due to its high expression in the intestine. Other MCT isoforms are less well characterized, but ongoing studies indicate that MCT6 transports xenobiotics such as bumetanide, nateglinide, and probenecid, whereas MCT7 has been characterized as a transporter of ketone bodies. MCT8 and MCT10 transport thyroid hormones, and recently, MCT9 has been characterized as a carnitine efflux transporter and MCT12 as a creatine transporter. Expressed at the blood brain barrier, MCT8 mutations have been associated with an X-linked intellectual disability, known as Allan-Herndon-Dudley syndrome. Many MCT isoforms are associated with hormone, lipid, and glucose homeostasis, and recent research has focused on their potential roles in disease, with MCTs representing promising novel therapeutic targets. This review will provide a summary of the current literature focusing on the characterization, function, and regulation of the MCT family isoforms and on their roles in drug disposition and in health and disease. SIGNIFICANCE STATEMENT: The 14-member solute carrier family 16 of monocarboxylate transporters (MCTs) plays a fundamental role in maintaining intracellular concentrations of a broad range of important endogenous molecules in health and disease. MCTs 1, 2, and 4 (L-lactate transporters) are overexpressed in cancers and represent a novel therapeutic target in cancer. Recent studies have highlighted the importance of MCTs in glucose, lipid, and hormone homeostasis, including MCT8 in thyroid hormone brain uptake, MCT12 in carnitine transport, and MCT11 in type 2 diabetes.

Effects of bisphenol S, a major substitute of bisphenol A, on neurobehavioral responses and cerebral monocarboxylate transporters expression in mice

Bisphenol A has been restricted in a large variety of products. Bisphenol S (BPS) is its major substitute. Yet, the impacts of BPS on the central nervous system are unknown, especially in vulnerable populations like children. The aim of this study was to investigate the effects of BPS on behavioral performances and the expression of cerebral monocarboxylate transporters (MCTs). Male Swiss mice were exposed to BPS at 100 μg/kg in drinking water for 10 weeks. The protocol started after the lactation period, which is a sensitive period of early social-emotional development. Elevated T-maze and open field tests were used to measure respectively, anxiety-related and activity-related behaviors. Molecular expressions of MCTs isoforms (MCT1, MCT2, MCT4) were determined in the frontal cortex. Data showed that BPS does not affect mRNA expression of MCTs. However, BPS decreases the number of entries into the open arms and the time spent on them for BPS-treated mice. These data reveal an anxiogenic effect of BPS. For locomotor activity and exploratory behavior levels, differences did not reach a statistically significant level in the BPS-exposed group. The effect of BPS on behavioral performances unravels a putative risk for psychopathology development in early childhood and calls for more attention.

Infusion of sodium butyrate promotes rumen papillae growth and enhances expression of genes related to rumen epithelial VFA uptake and metabolism in neonatal twin lambs.

The objective of this study was to evaluate the effect of sodium butyrate (SB) infusion on rumen papillae growth and volatile fatty acid (VFA) uptake and metabolism in neonatal lambs. Seven pairs of newborn twin lambs were used. Within each pair, lambs were assigned to receive an oral infusion of SB at 0.36 g/kg body weight (BW) (SB, n = 7) or the same volume of saline (Con, n = 7). Treatments were administered from 10 to 49 d of age, when all lambs were slaughtered. Results showed that the average daily feed intake (ADFI) of starter, average daily gain (ADG), BW of lambs at ages of 5 and 6 wk in SB group were greater (P < 0.05) than those in Con group. Infusion of SB increased (P < 0.05) the concentrations of acetate, butyrate, and total VFA in the rumen fluid and elevated (P < 0.05) the levels of β-hydroxybutyrate acid (BHBA), insulin-like growth factor-1 (IGF-1), and insulin in plasma. Infusion of SB promoted rumen papillae growth, depicted by higher emptied rumen weight, larger rumen papillae length, width, and surface area, and greater thickness of stratum corneum and total epithelium. Sodium butyrate infusion upregulated (P < 0.05) mRNA expression of cyclin A2, cyclin D1, and cyclin-dependent kinases 6 (CDK6), and downregulated (P < 0.05) mRNA expression of caspase-3 and Bcl-2-associated X protein (Bax) in the rumen epithelia. Moreover, SB infusion also upregulated (P < 0.05) mRNA expression of insulin-like growth factor-1 receptor (IGF-1R), and insulin-like growth factor-binding protein 5 (IGFBP-5), and downregulated (P < 0.05) mRNA expression of insulin-like growth factor-binding protein 3 (IGFBP-3) in the rumen epithelia. Sodium butyrate infusion also enhanced (P < 0.05) gene expressions of monocarboxylate transporter isoform 1 (MCT1), downregulated in adenoma (DRA), 3-hydroxy-3-methylglutaryl-CoA synthase isoform 2 (HMGCS2), and 3-hydroxy-3-methylglutaryl-CoA lyase (HMGCL), while depressed (P < 0.05) mRNA expression of sodium/proton exchanger isoform 2 (NHE2) in the rumen epithelia. Our results suggest that the SB infusion can improve animal performance, promote the ruminal papillae growth, and enhance expression of genes related to ruminal epithelial VFA uptake and metabolism in preweaning twin lambs. These findings provide a better understanding of the molecular mechanism of SB promoting rumen epithelial development and function in preweaning lambs.

Intense physical exercise potentiates glucose inhibitory effect over food intake of male Wistar rats.

What is the central question of this study? How does an acute session of exercise affect food intake of male Wistar rats? What is the main finding and its importance? Food intake in male Wistar rats is decreased in the first hour after physical exercise independent of the intensity. Moreover, high-intensity exercise potentiates the anorexic effect of peripheral glucose administration. This work raises new feeding-related targets that would explain how exercise drives body weight loss. Obesity has emerged as a critical metabolic disorder in modern society. An adequate lifestyle with a well-oriented programme of diet and physical exercise (PE) can prevent or potentially even cure obesity. Additionally, PE might lead to weight loss by increasing energy expenditure and decreasing hunger perception. In this article, we hypothesize that an acute exercise session would potentiate the glucose inhibitory effects on food intake in male Wistar rats. Our data show that moderate- or high-intensity PE significantly decreased food intake, although no changes in the expression of feeding-related neuropeptide in the arcuate nucleus of the hypothalamus were found. Exercised animals demonstrated a reduced glucose tolerance and increased blood insulin concentration. Intraperitoneal administration of glucose decreased food intake in control animals. In the animals submitted to moderate-intensity PE, the decrease in food intake promoted by glucose was similar to controls; however, an interaction was observed when glucose was injected in the high-intensity PE group, in which food intake was significantly lower than the effect produced by glucose alone. A different pattern of expression was observed for the monocarboxylate transporter isoforms (MCT1, 2 and 4) and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFBP3) in the hypothalamus, which was dependent on the exercise intensity. In conclusion, PE decreases food intake independently of the intensity. However, an interaction between PE and the anorexic effect of glucose is only observed when a high-intensity exercise is performed. These data show an essential role of exercise intensity in the modulation of the glucose inhibitory effect on food intake.

MCT1 regulates aggressive and metabolic phenotypes in bladder cancer.

Background: Monocarboxylate transporter isoform 1 (MCT1) is an important molecule in mediating lactate transportation. Recent studies have shown an oncogenic role of MCT1 in cancer development.Methods: In this study, we aimed to investigate the expression and role of MCT1 in bladder cancer (BCa). MCT1 expression was detected in 124 BCa tissues and their clinicopathological significance was analyzed. We also used The Cancer Genome Atlas database to explore the prognostic association of MCT1 with BCa. Cell proliferation, migration and invasion assays were performed on BCa cells in which MCT1 was downregulated. The effect of MCT1 on BCa cell aerobic glycolysis, as well as its association with HIF-1α, was tested.Results: We found that high MCT1 expression correlated with lymph node and distant metastasis. Patients with high-MCT1 expression showed shorter overall survival than those with low-MCT1 expression. Knockdown of MCT1 inhibited BCa cell proliferation, migration and invasion, and affected expression of epithelial-mesenchymal transition related proteins. Downregulation of MCT1 decreased lactate levels in cell medium, as well as HK2, GLUT1 and LDHB expression. In addition, MCT1 expression was partly dependent on HIF-1α.Conclusions: Taken together, our study has shown a prognostic role of MCT1 in BCa, and provided potential diagnostic and therapeutic options for BCa patients.

Effect of starter diet supplementation on rumen epithelial morphology and expression of genes involved in cell proliferation and metabolism in pre-weaned lambs

Starter feeding is usually used in lamb production to improve rumen development and to facilitate the weaning process, but molecular mechanism of which is not well understood. Therefore, the objective of this study is to investigate the effect of starter feeding on the expression of ruminal epithelial genes involved in cell proliferation, apoptosis and metabolism in pre-weaned lambs. We selected eight pairs of 10-day-old lamb twins. One twin was fed ewe milk (M, n=8), while the other was fed ewe milk plus starter (M+S, n=8). The lambs were sacrificed at 56 days age. Results showed that the lambs fed M+S had lower pH in the rumen and a higher concentration of acetate, propionate, butyrate and total volatile fatty acid (VFA). Compared with the M group, the concentration of β-hydroxybutyric acid in plasma had an increased trend, and the concentration of IGF-1 in plasma had an decreased trend in the M+S group. The length, width and surface of rumen papillae increased in the M+S group compared with the M group; this was associated with increased cell layers in the stratum corneum, stratum granulosum and total epithelia. Messenger RNA (mRNA) expression of proliferative genes of cyclin A, cyclin D1 and cyclin-dependent kinase 2 in the ruminal epithelia of M+S lambs was increased compared with M only lambs. The mRNA expression of apoptosis genes of caspase-3, caspase-8, B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in the M+S group was decreased compared with M group, but the ratio of Bcl-2 to Bax were not changed between the two groups. Expression of IGF-1 mRNA was decreased, but the mRNA expression of IGF-1 receptor was higher in ruminal epithelia in the M+S group. Furthermore, the mRNA expression of VFA absorption and metabolism genes of β-hydroxybutyrate dehydrogenase isoforms 1 and 3-hydroxy-3-methylglutaryl-CoA lyase had an increased trend in the M+S group than in the M group, but the mRNA expression of 3-hydroxy-3-methylglutaryl-CoA synthase isoform 1, monocarboxylate transporter isoform 1 and putative anion transporter isoform 1 had a decreased trend in the M+S group than in the M group. These results suggest that starter feeding increased proliferation and inhibited apoptosis of ruminal epithelial cells, and may promote the VFA metabolism in ruminal epithelium in pre-weaned lambs. These findings provide new insights into improving rumen development by nutritional intervention strategies in pre-weaned lambs.

Involvement of monocarboxylate transporter 1 (SLC16A1) in the uptake of l-lactate in human astrocytes

PurposeAstrocytes, the most abundant glial cells in the central nervous system (CNS), help neurons survive. Monocarboxylate transporters (MCTs) are reported to transport l-lactate, which is important for CNS physiology and cognitive function. However, it remains unclear which MCT isoform is functionally expressed by human astrocytes. The aim of this study was to establish the contribution of each MCT isoform to l-lactate transport in human astrocytes. MethodsThe function of l-lactate transport was studied using NHA cells as a human astrocyte model and radiolabeled l-lactate. The expression of MCT in human astrocytes was detected by immunohistochemistry staining. ResultsThe cellular uptake of l-lactate was found to be pH- and concentration-dependent with a Km value for l-lactate uptake of 0.64mM. This Km was similar to what has been previously established for MCT1-mediated l-lactate uptake. α-Cyano-4- hydroxycinnamate (CHC) and 5-oxoproline, which are both MCT1 inhibitors, were found to significantly inhibit the uptake of l-lactate, suggesting MCT1 is primarily responsible for l-lactate transport. Moreover, MCT1 protein was expressed in human astrocytes. ConclusionpH-dependent l-lactate transport is mediated by MCT1 in human astrocytes.

Monocarboxylate Transporters: Therapeutic Targets and Prognostic Factors in Disease.

Solute carrier (SLC) transporters represent 52 families of membrane transport proteins that function in endogenous compound homeostasis and xenobiotic disposition, and have been exploited in drug delivery and therapeutic targeting strategies. In particular, the SLC16 family thatencodes for the 14 isoforms of the monocarboxylate transporter (MCT) family plays a significant role in the absorption, tissue distribution, and clearance of both endogenous and exogenous compounds. MCTs are required for the transport of essential cell nutrients and for cellular metabolic and pH regulation. Recent publications have indicated their novel roles in disease, and thus their potential as biomarkers and new therapeutic targets in disease are under investigation. More research into MCT isoform function, specificity, expression, and regulation will allow researchers to exploit the potential utility of MCTs in the clinic as therapeutic targets and prognostic factors of disease.