Hyaluronic Acid Synthesis Research Articles

Effect of Epidermal Growth Factor-Like Peptides on the Metabolism of In Vitro- Matured Mouse Oocytes and Cumulus Cells1

Oocyte in vitro maturation (IVM) is an assisted reproductive technology that involves the maturation of cumulus-oocyte complexes (COCs) that are then capable of normal development. We have shown that epidermal growth factor (EGF)-like peptide signaling is perturbed in mouse COCs undergoing IVM when matured with follicle-stimulating hormone (FSH) and/or EGF, but supplementation of IVM with EGF-like peptides amphiregulin or epiregulin improves oocyte developmental competence. Here we aimed to determine whether EGF-like peptides regulate COC metabolism. Immature 129/Sv mouse COCs underwent IVM with FSH, EGF, amphiregulin, epiregulin, betacellulin, or no treatment (control). Epiregulin significantly increased intraoocyte flavin adenine dinucleotide (FAD) and REDOX (reduction and oxidation) ratio compared to FSH and control. Amphiregulin and epiregulin significantly increased the proportion of J aggregates (from JC-1) in oocyte mitochondria compared to control, FSH, or EGF, and this coupled with FAD and REDOX measures indicates greater mitochondrial activity. There were no differences in glucose consumption, lactate production, or glycolysis between COCs matured with FSH, EGF, and EGF-like peptides. COCs matured with EGF or EGF-like peptides exhibited significantly higher mRNA expression of the hexosamine biosynthesis pathway (HBP) rate-limiting enzyme gene Gfpt2, Has2 expression, and global beta-O-linked glycosylation of proteins, compared to control or FSH, suggesting greater HBP activity. Our findings suggest that 1) EGF-like peptides, particularly epiregulin, induce more oocyte mitochondrial activity than EGF or FSH and 2) EGF-like peptides and EGF induce greater HBP activity, enabling more hyaluronic acid synthesis and protein beta-O-linked glycosylation. These metabolic alterations may be a mechanism by which EGF-like peptides increase oocyte developmental competence.

Cyclic phosphatidic acid relieves osteoarthritis symptoms.

BackgroundCyclic phosphatidic acid (cPA) is a naturally occurring phospholipid mediator with a unique cyclic phosphate ring at the sn-2 and sn-3 positions of its glycerol backbone. Natural cPA and its chemically stabilized cPA derivative, 2-carba-cPA (2ccPA), inhibit chronic and acute inflammation, and 2ccPA attenuates neuropathic pain. Osteoarthritis (OA) is a degenerative disease frequently associated with symptoms such as inflammation and joint pain. Because 2ccPA has obvious antinociceptive activity, we hypothesized that 2ccPA might relieve the pain caused by OA. We aimed to characterize the effects of 2ccPA on the pathogenesis of OA induced by total meniscectomy in the rabbit knee joint.ResultsIntra-articular injection of 2ccPA (twice a week for 42 days) significantly reduced pain and articular swelling. Histopathology showed that 2ccPA suppressed cartilage degeneration in OA. We also examined the effects of 2ccPA on the inflammatory and catabolic responses of human OA synoviocytes and chondrosarcoma SW1353 cells in vitro. 2ccPA stimulated synthesis of hyaluronic acid and suppressed production of the metalloproteinases MMP-1, -3, and -13. However, it had no effect on the production of interleukin (IL)-6, an inflammatory cytokine. The suppressive effect of 2ccPA on MMP-1 and -3 production in synoviocytes and on MMP-13 production in SW1353 cells was not mediated by the lysophosphatidic acid receptor, LPA1 receptor (LPA1R).ConclusionsOur results suggest that 2ccPA significantly reduces the pain response to OA by inducing hyaluronic acid production and suppressing MMP-1, -3, and -13 production in synoviocytes and chondrocytes.

Inhibition of Hyaluronan Synthase 2 Reduces the Abnormal Migration Rate of Keloid Keratinocytes

Keloids are fibroproliferative scars that spread beyond the original wound boundary and are very resistant to treatment. Development of highly effective therapies requires a comprehensive understanding of the mechanisms regulating keloid formation. Previous studies indicated that keloid keratinocytes have abnormal expression of genes involved in differentiation and adhesion, and increased migration rates. The objective of the current study was to better understand the role of hyaluronan synthase 2 (HAS2) in keloid keratinocyte migration and gene expression. Keratinocytes were isolated from keloid scars and normal skin. Migration rates of keloid keratinocytes were quantified using an in vitro scratch assay. Expression levels of HAS2, related HAS1, and HAS3 genes, and genes aberrantly expressed in keloid keratinocytes, were quantified using real-time polymerase chain reaction. Treatment with 4-methylumbelliferone (4MU) was used to inhibit hyaluronic acid synthesis. The expression of HAS2 was significantly increased in keloid vs normal keratinocytes. Treatment with 4MU caused a dose-dependent reduction in keloid keratinocyte migration and HAS2 expression; HAS3 expression was moderately inhibited by 4MU and HAS1 was not expressed. Keloid keratinocytes displayed a motile phenotype in vitro, including loose colonies and widely separated refractile cells; this phenotype was normalized by 4MU. Further, 4MU altered gene expression in keloid keratinocytes. The results suggest that HAS2 overexpression contributes to increased migration and altered gene expression in keloid keratinocytes. Abnormal keratinocyte migration may contribute to the overhealing of keloid scars beyond the original wound boundaries. Therefore, inhibition of HAS2 expression using 4MU may represent a novel strategy for treatment of keloid scarring.

천궁으로부터 분리된 ferulic acid의 히알루론산 생성에 미치는 효과

Hyaluronic acid (HA) is one of the major extracellular matrix components in skin. The HA content is reportedto decline with age, which may contribute to decrease in skin moisture, wrinkle formation and the decrease in elasticityof the skin. Among the family of HA synthase genes (HAS-1, 2, 3) identified so far, HAS-2 plays crucial roles inthe regulation of HA synthesis in human skin fibroblasts. In this study, we elucidated the effects of ferulic acid isolatedfrom Cnidium officinale on HA production. Semi-quantitative RT-PCR and quantitative real-time PCR showed that ferulicacid increased mRNA level of HAS-2 gene and ELISA assay also revealed that ferulic acid increased HA productionin human skin fibroblasts. Our study suggests that ferulic acid might prevent age-dependent skin deteriorations suchas wrinkles, dryness and elasticity decrease, all of which could be ascribed to the reduction of the HA content in humanskin.

RocA Truncation Underpins Hyper-Encapsulation, Carriage Longevity and Transmissibility of Serotype M18 Group A Streptococci

Group A streptococcal isolates of serotype M18 are historically associated with epidemic waves of pharyngitis and the non-suppurative immune sequela rheumatic fever. The serotype is defined by a unique, highly encapsulated phenotype, yet the molecular basis for this unusual colony morphology is unknown. Here we identify a truncation in the regulatory protein RocA, unique to and conserved within our serotype M18 GAS collection, and demonstrate that it underlies the characteristic M18 capsule phenotype. Reciprocal allelic exchange mutagenesis of rocA between M18 GAS and M89 GAS demonstrated that truncation of RocA was both necessary and sufficient for hyper-encapsulation via up-regulation of both precursors required for hyaluronic acid synthesis. Although RocA was shown to positively enhance covR transcription, quantitative proteomics revealed RocA to be a metabolic regulator with activity beyond the CovR/S regulon. M18 GAS demonstrated a uniquely protuberant chain formation following culture on agar that was dependent on excess capsule and the RocA mutation. Correction of the M18 rocA mutation reduced GAS survival in human blood, and in vivo naso-pharyngeal carriage longevity in a murine model, with an associated drop in bacterial airborne transmission during infection. In summary, a naturally occurring truncation in a regulator explains the encapsulation phenotype, carriage longevity and transmissibility of M18 GAS, highlighting the close interrelation of metabolism, capsule and virulence.

Hyaluronic Acid: From Biochemical Characteristics to its Clinical Translation in Assessment of Liver Fibrosis

Context:Hyaluronic acid (HA) is a high molecular weight polysaccharide that is distributed in all bodily tissues and fluids. The liver is the most important organ involved in the synthesis and degradation of HA. Research has shown that liver cell injury can affect serum HA levels. In this review, authors aimed to describe the biochemical and physiological roles of this glycosaminoglycan and its changes in various liver diseases.Evidence Acquisition:Liver fibrosis and in more severe form, cirrhosis are results of an imbalance between fibrogenesis and fibrinolysis. Liver biopsy is the gold standard to assess liver necro inflammatory injuries. This method is invasive and has some major side effects; therefore it is an unfavorable method for both physicians and patients. Now, a wide variety of noninvasive methods have been introduced based on evaluating serum level of different markers. They are safe, readily available, and more favorable. Serum HA levels are used by some researchers to assess stages of liver fibrosis.Results:There are several scientific studies indicating HA as a biomarker for high score fibrosis and cirrhosis in various liver diseases alone or in algorithm models. It seems from various algorithm models that the use of HA as a major constituent has more diagnostic reliability and accuracy than the use of HA alone.Conclusions:Use of HA in an algorithm model, is an extra and valuable tool for assessing liver necro inflammatory injuries- in parallel with liver biopsy- but more comprehensive studies are needed to approve the use of HA as an appropriate clinical tool.

High molecular weight hyaluronic acid regulates osteoclast formation by inhibiting receptor activator of NF-κB ligand through Rho kinase

To determine the effects of high molecular weight hyaluronic acid (HMW-HA) on osteoclast differentiation by monocytes co-cultured with stromal cells. Mouse bone marrow stromal cell line ST2 cells were incubated with HMW-HA or 4-methylunbeliferone (4-MU) for various times. In some experiments, cells were pre-treated with the anti-CD44 monoclonal antibody (CD44 mAb) or Rho kinase pathway inhibitors (simvastatin or Y27632), then treated with HMW-HA. The expression of receptor activator of NF-κB ligand (RANKL) was determined using real-time reverse transcription polymerase chain reaction (RT-PCR), western blotting, and immunofluorescence microscopy, while the amount of active RhoA was measured by a pull-down assay. To further clarify the role of HMW-HA in osteoclastogenesis, mouse monocyte RAW 264.7 cells were co-cultured with ST2 cells pre-stimulated with 1,25(OH)2D3. Osteoclast-like cells were detected by staining with tartrate-resistant acid phosphatase (TRAP). HMW-HA decreased RANKL mRNA and protein expressions, whereas inhibition of hyaluronic acid (HA) synthesis by 4-MU enhanced RANKL expression. Blockage of HA-CD44 binding by CD44 mAb suppressed HMW-HA-mediated inhibition of RANKL. Pull-down assay findings also revealed that HMW-HA transiently activated RhoA in ST2 cells and pre-treatment with CD44 mAb inhibited the activation of RhoA protein mediated by HMW-HA. Moreover pre-treatment with Rho kinase pathway inhibitors also blocked the inhibition of RANKL by HMW-HA. Co-culture system results showed that HMW-HA down-regulated differentiation into osteoclast-like cells by RAW 264.7 cells induced by 1,25(OH)2D3-stimulated ST2 cells. These results indicated that HA-CD44 interactions down-regulate RANKL expression and osteoclastogenesis via activation of the Rho kinase pathway.

Inhibition of adipogenesis in 3T3-L1 cells and suppression of abdominal fat accumulation in high-fat diet-feeding C57BL/6J mice after downregulation of hyaluronic acid

Adipogenesis can be spatially and temporally regulated by extracellular matrix (ECM). We hypothesized that the regulation of hyaluronic acid (HA), a component of the ECM, can affect adipogenesis in fat cells. The effects of HA on adipogenesis were investigated in vitro in 3T3-L1 cells and in vivo in high-fat diet-feeding C57BL/6J mice. We investigated the effects of HA by degradation of pre-existing or synthesized HA and artificial inhibition of HA synthesis in adipogenesis. In vitro adipogenesis in 3T3-L1 cells was inhibited by treating them with exogenous hyaluronidase (HYAL) and with 4-methylumbelliferone, which inhibited the synthesis of HA in a concentration-dependent manner. In vivo, abdominal fat accumulation in high-fat diet-feeding C57BL/6J mice was suppressed by exogenous HYAL 10(4) IU injections, which was associated with reduction of lipid accumulation in liver and increase of insulin sensitivity. Changes in the ECM such as accumulation of high molecular weight of HA by HAS and degradation of HA by endogenous HYAL were essential for adipogenesis both in vitro and in vivo.

Treatment with the hyaluronic acid synthesis inhibitor 4-methylumbelliferone suppresses SEB-induced lung inflammation.

Exposure to bacterial superantigens, such as staphylococcal enterotoxin B (SEB), can lead to the induction of acute lung injury/acute respiratory distress syndrome (ALI/ARDS). To date, there are no known effective treatments for SEB-induced inflammation. In the current study we investigated the potential use of the hyaluronic acid synthase inhibitor 4-methylumbelliferone (4-MU) on staphylococcal enterotoxin B (SEB) induced acute lung inflammation. Culturing SEB-activated immune cells with 4-MU led to reduced proliferation, reduced cytokine production as well as an increase in apoptosis when compared to untreated cells. Treatment of mice with 4-MU led to protection from SEB-induced lung injury. Specifically, 4-MU treatment led to a reduction in SEB-induced HA levels, reduction in lung permeability, and reduced pro-inflammatory cytokine production. Taken together, these results suggest that use of 4-MU to target hyaluronic acid production may be an effective treatment for the inflammatory response following exposure to SEB.

Formation of synovial joints and articular cartilage

Chondrocytes differentiate from mesenchymal progenitors and produce templates(anlagen) for the developing bones. Chondrocyte differentiation is controlled by Sox transcription factors. Templates for the neighbour bones are subsequently separated by conversion of differentiated chondrocytes into non-chondrogenic cells and emergence of interzone in which joints cavitation occurs. A central role in initiating synovial joint formation plays Wnt-14/beta-catenin signalling pathway.Moreover, bone morphogenetic proteins and growth and differentiation factors are expressed at the site of joint formation. Joint cavitation is associated with increased hyaluronic acid synthesis. Hyaluronic acid facilitates tissue separation and creation of a functional joint cavity. According to the traditional view articular cartilage represents part of cartilage anlage that is not replaced by bone through endochondral ossification. Recent studies indicate, however, that peri-joint mesenchymal cells take part in interzone formation and that these interzone cells subsequently differentiate into articular chondrocytes and synovial cells. Thus,anlage chondrocytes have a transient character and disappear after cessation of growth plate function while articular chondrocytes have stable and permanent phenotype and function throughout life.

Synthesis and Use of Hyaluronic Acid–10B Polymeric Chelates for Neutron-Capture Therapy

The technology of solid-state modification of hyaluronic acid (HA) with sodium decahydroborate by deformation in Bridgman anvils was developed. The deformation was carried out at a pressure of 1 GPa and a rotation angle of 500°. The structure of the obtained complexes was studied. It was shown using Raman and IR-Fourier spectroscopy that HA and the polyborates could form a network of cyclic polymeric chelate complexes in which HA acted as a multidentate ligand. The biodistribution of the HA-based B-containing compound in organs and tissues of mice with B-16 melanoma was studied after a single intratumoral administration. The maximum B content in the tumor was observed 15 min after administration. However, it was halved after 1 h and decreased by 5 times by 3 h. The most opportune time for performing a boron-neutron-capture therapy session was 30 min after administration when the tumor B concentration was >30 μg/g and exceeded the content in surrounding tissues. The tumor/muscle and tumor/blood ratios reached 3 and 5, respectively. Use of this non-toxic compound for neutron-capture therapy was deemed promising based on preclinical studies.

The Role of Hyaluronic Acid Precursor Concentrations in Molecular Weight Control in Streptococcus zooepidemicus

The biosynthetic pathway responsible for the production of hyaluronic acid (HA) has been thoroughly studied; however, many aspects remain elusive regarding the mechanisms that control molecular weight (MW). Previously, we demonstrated a positive correlation between MW and the concentration of the HA precursor sugar UDP-N acetylglucosamine (UDP-GlcNAc). To further investigate the role of UDP-GlcNAc in MW control, we increased the intracellular concentration of this monomer using both feeding strategies and genetic engineering approaches. Feeding cells glucosamine dramatically increased intracellular levels of UDP-GlcNAc, but unexpectedly, produced HA of a lower MW. This was subsequently attributed to an equally dramatic decrease in the level of the other HA precursor sugar UDP-glucuronic acid (UDP-GlcUA). Feeding cells a mixture of glucose and GlcNAc addressed this imbalance of precursor sugars, leading to an increase in both UDP-GlcNAc and UDP-GlcUA; however, no significant increase in MW was observed. Despite the increase in UDP-sugars, RNA sequencing identified no increase in the expression of the genes involved in production of HA. Returning to genetic engineering approaches to examine UDP-GlcNAc and MW, genes known to contribute to the production of UDP-GlcNAc were over-expressed, both individually and together. Using this approach, UDP-GlcNAc and MW increased. At lower levels of UDP-GlcNAc, the positive correlation between UDP-GlcNAc levels and MW was maintained, however this relationship stalled at higher concentrations of UDP-GlcNAc. Taken together, these results suggest that while optimising HA precursor levels using feeding or genetic engineering approaches can improve HA MW, there is a point at which excess availability of precursors is no longer advantageous. Once precursor concentrations are addressed, it would seem that other uncharacterised factor(s) (e.g. rate of HA synthesis) also contribute to HA MW control.

Targeting hyaluronic acid production for the treatment of leukemia: Treatment with 4-methylumbelliferone leads to induction of MAPK-mediated apoptosis in K562 leukemia

The current study examined the effect of modulation of hyaluronic acid (HA) synthesis on leukemia cell survival using the hyaluronic acid synthesis inhibitor 4-methylumbelliferone (4-MU). Treatment of CML cells with 4-MU led to caspase-dependent apoptosis characterized by decreased HA production, PARP cleavage, and increased phosphorylation of p38. Addition of exogenous HA, the pan caspase inhibitor Z-VAD-FMK or the p38 inhibitor SB203580 to 4-MU treated cells was able to protect cells from apoptosis. Treatment of tumor-bearing mice with 4-MU led to a significant reduction in tumor load which was mediated through the induction of apoptosis.

Glycosaminoglycan Regulation by VEGFA and VEGFC of the Glomerular Microvascular Endothelial Cell Glycocalyx in Vitro

Damage to endothelial glycocalyx impairs vascular barrier function and may contribute to progression of chronic vascular disease. An early indicator is microalbuminuria resulting from glomerular filtration barrier damage. We investigated the contributions of hyaluronic acid (HA) and chondroitin sulfate (CS) to glomerular microvascular endothelial cell (GEnC) glycocalyx and examined whether these are modified by vascular endothelial growth factors A and C (VEGFA and VEGFC). HA and CS were imaged on GEnCs and their resynthesis was examined. The effect of HA and CS on transendothelial electrical resistance (TEER) and labeled albumin flux across monolayers was assessed. Effects of VEGFA and VEGFC on production and charge characteristics of glycosaminoglycan (GAG) were examined via metabolic labeling and liquid chromatography. GAG shedding was quantified using Alcian Blue. NDST2 expression was examined using real-time PCR. GEnCs expressed HA and CS in the glycocalyx. CS contributed to the barrier to both ion (TEER) and protein flux across the monolayer; HA had only a limited effect. VEGFC promoted HA synthesis and increased the charge density of synthesized GAGs. In contrast, VEGFA induced shedding of charged GAGs. CS plays a role in restriction of macromolecular flux across GEnC monolayers, and VEGFA and VEGFC differentially regulate synthesis, charge, and shedding of GAGs in GEnCs. These observations have important implications for endothelial barrier regulation in glomerular and other microvascular beds.

AB0066 Cyclic phosphatidic acid (cpa) stimulates hyaluronic acid production in human osteoarthritic articular chondrocytes, and intra-articular administration of cpa suppresses pain, swelling, and cartilage destruction in rabbit experimental osteoarthritis.

BackgroundCyclic phosphatidic acid (cPA) is one of lipid mediators, has been shown various biological effects1. On human skin fibroblasts, cPA stimulates high molecular hyaluronic acid (HA) production through up-regulating HA...

AB0083 Anti-inflammatory effect of hymovis® on an in vitro model of CPP crystal-induced inflammation. Comparison with other hyaluronic acids used for intra-articular treatment

BackgroundIntra-articular injections of hyaluronic acid (HA) are widely used in the treatment of inflammatory and degenerative joint diseases. A novel slightly modified hyaluronic acid, HYADD4 (Hymovis®), has recently been tested...

Altered expression of hyaluronan synthase and hyaluronidase mRNA may affect hyaluronic acid distribution in keloid disease compared with normal skin

Keloid disease (KD) is a fibroproliferative disorder characterised partly by an altered extracellular matrix (ECM) profile. In fetal scarring, hyaluronic acid (HA) expression is increased, but is reduced in KD tissue compared with normal skin (NS). The expression of Hyaluronan Synthase (HAS) and hyaluronidase (HYAL) in KD and NS tissue were investigated for the first time using a range of techniques. Hyaluronan synthase and HYAL mRNA expression were significantly increased in NS tissue compared with KD tissue (P < 0.05). Immunohistological analysis of tissue indicated an accumulation of HAS and HYAL protein expression in KD compared with NS due to the thicker epidermis. No differences were observed in mRNA or protein expression in KD and NS fibroblasts. Reduced expression of HAS and HYAL may alter HA synthesis, degradation and accumulation in KD. Better understanding of the role of HA in KD may lead to novel therapeutic approaches to address the resulting ECM imbalance.

Vitamin D inhibition of Has2 and hyaluronan synthesis in invasive breast cancer cells

1,25‐Dihydroxyvitamin D (1,25D) exerts anti‐cancer actions through the vitamin D receptor (VDR) but the specific targets that mediate these effects remain to be defined. In this study, growth and genomic responses to 1,25D were evaluated in cells derived from mammary tumors generated in VDR knock‐out (KO) and wild‐type (WT) mice. WT145 cells (derived from WT tumors) expressed VDR and were growth inhibited by 1,25D, whereas KO240 cells (derived from VDRKO tumors) lacked VDR and were not growth inhibited by 1,25D. KO240 cell clones stably expressingVDR (KOhVDR cells) were sensitized to 1,25D mediated growth arrest. Genomic profiling of KO240, KOhVDR and WT145 cells identified 35 transcripts that were altered by 1,25D in relation to growth inhibition, with an enrichment of genes involved in heparin sulfate and proteoglycan binding (including Prelp, Hbegf, Fbn1, Postn and Has2). Currently we are focusing on VDR regulation of Has2 in mammary gland and breast cancer. Has2 is the rate‐limiting enzyme in the synthesis of hyaluronic acid (HA) which is strongly associated with tumor invasion and metastasis. To date, we have confirmed down‐regulation of Has2 protein by 1,25D and demonstrated enhanced HA accumulation in mammary gland and tumors of VDRKO mice. These data define a novel role for vitamin D in control of HA synthesis in epithelial tissues that likely contributes to its anti‐cancer actions.NIH R01CA69700

Dermial®: A new natural ingredient with anti‐aging and regenerative properties for skin

ObjectiveTo determine the anti‐aging and regenerative effect of Dermial®, a new ingredient with Cosmetic and Nutricosmetic potential.Dermial® is a natural product rich in glycosaminoglycans, mainly hyaluronic acid, and also type I collagen, both beneficial for skin health.MethodsThe anti‐aging action was determined using a culture of human dermal fibroblasts (HDF). Skin hydration effect was quantified by measuring the synthesis of hyaluronic acid. The potential to increase skin elasticity was determined quantifying elastin levels. Effects on proliferation and migration of HDF were also tested.ResultsDermial® showed a hydrating effect at the 3 tested concentrations in a dose‐dependent manner. This nutricosmetic ingredient was able to increase skin moisture up to 30% and to induce a statistical significant effect on the elastin synthesis up to 16%. In addition, this new ingredient was a potent inductor of fibroblast migration and was also able to increase up to 100% the proliferation index compared to the baseline culture.ConclusionsThese data demonstrate that Dermial® exhibits antiaging properties due to its hydrating action and its stimulating effect of elastin production. Moreover, it shows a powerful regenerative activity. All together, these results suggest that Dermial® may be useful as an anti‐aging treatment.Supported by BIOIBERICA S.A., Barcelona, Spain.

168 DIETARY SUPPLEMENT HYMECROMONE BASED COMBINATION FOR RENAL CELL CARCINOMA

You have accessJournal of UrologyKidney Cancer: Basic Research (I)1 Apr 2013168 DIETARY SUPPLEMENT HYMECROMONE BASED COMBINATION FOR RENAL CELL CARCINOMA Nicolas Ortiz, Anaid Benitez, Travis Yates, and Vinata Lokeshwar Nicolas OrtizNicolas Ortiz Miami, FL More articles by this author , Anaid BenitezAnaid Benitez Miami, FL More articles by this author , Travis YatesTravis Yates Miami, FL More articles by this author , and Vinata LokeshwarVinata Lokeshwar Miami, FL More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.1548AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Approximately 17% of patients with renal cell carcinoma (RCC) present with metastasis upon diagnosis, rendering patients with a 5-year survival rate of 12%. Sorafenib (SF) was the first targeted therapy approved for RCC treatment, however it has modest efficacy. Hyaluronic acid (HA) is a non-sulfated glycosaminoglycan that promotes cell proliferation, invasion and motility through membrane receptors CD44 and RHAMM. High HA expression has been correlated with cancer progression and inflammatory diseases. Hymecromone (HC) is a non−toxic dietary supplement that inhibits HA synthesis and down regulates CD44 and RHAMM mRNA. In this study we examined whether HC, SF, or their combination inhibit RCC progression. METHODS Effect of HC, SF and HC+SF on cell proliferation, cell cycle and apoptosis was examined in RCC cells (Caki−1, 786−O) and endothelial cells (HMVEC−L, HUVEC) by cell-counting, flow cytometry and Cell Death ELISA. Boyden chamber was used for motility and invasion assays. Effect on cell cycle, apoptosis, and HA receptor levels were evaluated by immunoblotting and Q−PCR. Treatment efficacy was evaluated in the Caki−1 xenograft model. RESULTS While HC (0−30 ug/ml) and SF (0−3.2 ug/ml) individually were ineffective, their combination inhibited proliferation (>95%), motility (≥60%) and invasion (≥50%) in both RCC and endothelial cells. Combination of HC+ SF induced apoptosis (>8−fold) and caused cell cycle arrest. Consistent with these results, downregulation of Cyclins A, E1, D1, phospho−cdk2, Rb, STAT3, Mcl−1 and induced PARP cleavage, and Caspase 3 activation was also seen in combination treatment. HC+SF synergistically decreased the expression of HA receptors CD44 and RHAMM at the mRNA and protein levels. The oral administration of both HC (50−100mg/kg) and SF (30mg/kg) eradicated SF-resistant Caki−1 tumors whereas individual drugs were ineffective. No serum or tissue toxicity was observed in the treated groups. CONCLUSIONS Our findings show that a novel combination of SF with HC, a non−toxic oral supplement is potentially an effective treatment strategy for metastatic RCC. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e69 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Nicolas Ortiz Miami, FL More articles by this author Anaid Benitez Miami, FL More articles by this author Travis Yates Miami, FL More articles by this author Vinata Lokeshwar Miami, FL More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...