Marine Sponge Haliclona Research Articles

Potential of Polar Lipids Isolated from the Marine Sponge Haliclona (Halichoclona) vansoesti against Melanoma.

Marine sponges represent a good source of natural metabolites for biotechnological applications in the pharmacological, cosmeceutical, and nutraceutical fields. In the present work, we analyzed the biotechnological potential of the alien species Haliclona (Halichoclona) vansoesti de Weerdt, de Kluijver & Gomez, 1999, previously collected in the Mediterranean Sea (Faro Lake, Sicily). The bioactivity and chemical content of this species has never been investigated, and information in the literature on its Caribbean counterpart is scarce. We show that an enriched extract of H. vansoesti induced cell death in human melanoma cells with an IC50 value of 36.36 µg mL-1, by (i) triggering a pro-inflammatory response, (ii) activating extrinsic apoptosis mediated by tumor necrosis factor receptors triggering the mitochondrial apoptosis via the involvement of Bcl-2 proteins and caspase 9, and (iii) inducing a significant reduction in several proteins promoting human angiogenesis. Through orthogonal SPE fractionations, we identified two active sphingoid-based lipid classes, also characterized by nuclear magnetic resonance and mass spectrometry, as the main components of two active fractions. Overall, our findings provide the first evaluation of the anti-cancer potential of polar lipids isolated from the marine sponge H. (Halichoclona) vansoesti, which may lead to new lead compounds with biotechnological applications in the pharmaceutical field.

Gelatin nanoparticles loaded with 3-alkylpyridinium salt APS7, an analog of marine toxin, are a promising support in human lung cancer therapy

This study demonstrates the potential of gelatin nanoparticles as a nanodelivery system for antagonists of nicotinic acetylcholine receptors (nAChRs) to improve chemotherapy efficacy and reduce off-target effects. Too often, chemotherapy for lung cancer does not lead to satisfactory results. Therefore, new approaches directed at multiple pharmacological targets in cancer therapy are being developed. Following the activation of nAChRs (e.g. by nicotine), cancer cells begin to proliferate and become more resistant to chemotherapy-induced apoptosis. This work shows that the 3-alkylpyridinium salt, APS7, a synthetic analog of a toxin from the marine sponge Haliclona (Rhizoneira) sarai, acts as an nAChR antagonist that inhibits the pro-proliferative and anti-apoptotic effects of nicotine on A549 human lung adenocarcinoma cells. In this study, gelatin-based nanoparticles filled with APS7 (APS7-GNPs) were prepared and their effects on A549 cells were compared with that of free APS7. Both APS7 and APS7-GNPs inhibited Ca2+ influx and increased the efficacy of cisplatin chemotherapy in nicotine-stimulated A549 cells. However, significant benefits from APS7-GNPs were observed – a stronger reduction in the proliferation of A549 lung cancer cells and a much higher selectivity in cytotoxicity towards cancer cells compared with non-tumorigenic lung epithelial BEAS-2B cells.

Marine Natural Compound (Neviotin A) Displays Anticancer Efficacy by Triggering Transcriptomic Alterations and Cell Death in MCF-7 Cells.

We investigated the anticancer mechanism of a chloroform extract of marine sponge (Haliclona fascigera) (sample C) in human breast adenocarcinoma (MCF-7) cells. Viability analysis using MTT and neutral red uptake (NRU) assays showed that sample C exposure decreased the proliferation of cells. Flow cytometric data exhibited reactive oxygen species (ROS), nitric oxide (NO), dysfunction of mitochondrial potential, and apoptosis in sample C-treated MCF-7 cells. A qPCR array of sample C-treated MCF-7 cells showed crosstalk between different pathways of apoptosis, especially BIRC5, BCL2L2, and TNFRSF1A genes. Immunofluorescence analysis affirmed the localization of p53, bax, bcl2, MAPKPK2, PARP-1, and caspase-3 proteins in exposed cells. Bioassay-guided fractionation of sample C revealed Neviotin A as the most active compound triggering maximum cell death in MCF-7, indicating its pharmacological potency for the development of a drug for the treatment of human breast cancer.

Antiproliferative and pro-apoptotic properties against colon carcinoma cell line of the marine sponge haliclona sp. From poblacion, kauswagan, lanao del norte, Philippines

Marine sponge Haliclona sp. was examined for its anticancer potential focusing on the antiproliferative and pro-apoptotic properties against HCT116 (colon) cancer cell line. The 16 fractions obtained from Haliclona sp. were subjected to antiproliferative and pro-apoptotic evaluation via MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and Caspase-Glo 3/7 assay, respectively, at 30 μg/mL concentration. Fractions 10, 13, 14, and 15 exhibited a significant decrease (p < 0> 50�ll viability) against HCT116. Only fraction 8 showed a significant increase (p < 0.05) in relative light unit (RLU) and fold change (FC) (1809.78 and 2.42), more than two times larger, compared to the negative control (754.89 RLU and 1.00 FC). This result demands further study on fraction 8 to identify and characterize the structure of the compounds responsible for its bioactivity and hence could be a source of drug leads.

Purification and biochemical characterization of SM14est, a PET-hydrolyzing enzyme from the marine sponge-derived Streptomyces sp. SM14.

The successful enzymatic degradation of polyester substrates has fueled worldwide investigation into the treatment of plastic waste using bio-based processes. Within this realm, marine-associated microorganisms have emerged as a promising source of polyester-degrading enzymes. In this work, we describe the hydrolysis of the synthetic polymer PET by SM14est, a polyesterase which was previously identified from Streptomyces sp. SM14, an isolate of the marine sponge Haliclona simulans. The PET hydrolase activity of purified SM14est was assessed using a suspension-based assay and subsequent analysis of reaction products by UV-spectrophotometry and RP-HPLC. SM14est displayed a preference for high salt conditions, with activity significantly increasing at sodium chloride concentrations from 100 mM up to 1,000 mM. The initial rate of PET hydrolysis by SM14est was determined to be 0.004 s-1 at 45°C, which was increased by 5-fold to 0.02 s-1 upon addition of 500 mM sodium chloride. Sequence alignment and structural comparison with known PET hydrolases, including the marine halophile PET6, and the highly efficient, thermophilic PHL7, revealed conserved features of interest. Based on this work, SM14est emerges as a useful enzyme that is more similar to key players in the area of PET hydrolysis, like PHL7 and IsPETase, than it is to its marine counterparts. Salt-tolerant polyesterases such as SM14est are potentially valuable in the biological degradation of plastic particles that readily contaminate marine ecosystems and industrial wastewaters.

New lectin isolated from the tropical sponge Haliclona (Reniera) implexiformis (Hechtel, 1965) shows antibiofilm effect

A lectin from the marine sponge Haliclona (Reniera) implexiformis (HiL) was isolated by affinity chromatography on Sepharose™ matrix. HiL showed specificity for galactose and its derivatives. The glycoproteins porcine stomach mucin (PSM) and bovine stomach mucin (BSM) were potent inhibitors. Hemagglutinating activity of the lectin was maximal between pH 5.0 and 9.0. The lectin remained active until 60°C. The presence of CaCl2 and EDTA did not affect the hemagglutinating activity. In SDS-PAGE, HiL showed a single band of 20 kDa under reduced conditions, whereas in the non-reducing conditions, it showed a band of 20 kDa and one additional band of 36 kDa. The average molecular mass determined by Electrospray Ionization Mass Spectrometry (ESI-MS) was 35.874 ± 2 Da in native and non-reducing conditions, whereas carboxyamidomethylated-lectin showed 18,111 Da. These data indicated that HiL consists in a dimer formed by identical subunits linked by disulfide bonds. Partial amino acid sequence of HiL was determined by mass spectrometry, and revealed that it is a new type of lectin, which showed no similarity with any protein. Secondary structure consisted of 6% α-helice, 31% β-sheet, 18% β-turn and 45% random coil. HiL showed significant reduction in the number of viable cells of Staphylococcus biofilms.

Large-scale mariculture of the sponge Haliclona simulans on floating rafts in Zhao'an Bay, Fujian Province, China

Marine sponges are an important source of bioactive products and biomaterials. However, there is a bottleneck of biomass supply for sponge utilization. To overcome this bottleneck, mariculture was considered to be the most viable solution. Mariculture of the marine sponge Haliclona simulans was conducted on floating rafts in Zhao'an Bay, Fujian Province, China. The explants were suspended 0.5–2 m below the water surface to grow naturally using the mesh method. Two independent trials with different scales were performed in this study. In a small-scale trial with 30 sponge explants, with daily maintenance, the survival rate of H. simulans was 93.3% over 224 days. To determine whether harvesting twice yearly yields greater output than annual harvesting, the 30 H. simulans were randomly divided into two equivalent groups. The annual harvest group (1 harvest per year, 1PY) was harvested in 224 days, and the twice harvest groups (2 harvest per year, 2PY) were harvested at 92 days and 224 days, with approximately 30% of the biomass was retained during the first harvest to allow regrowth. The results showed that the total biomass accumulation rate was 629.27 ± 256.74% for the 1PY group and 1014.73 ± 252.56% for the 2PY group. The biomass accumulation rate of 2PY was approximately 61.26% higher than that of 1PY. To obtain more accurate yield data under industry scale, a large-scale mariculture was independently conducted with about 7500 explants and covered a sea area about 500 m2, in which each area of 0.2 m2 was occupied by a horizontal rope, and each rope held 3 vertically suspended explants. Lost or dead explants were replanted during routine maintenance. A total of 330 kg wet weight biomass was obtained from the first harvest after 92 days of cultivation, and 272 kg wet weight biomass was obtained from the second harvest after 224 days. A total of 45.05 kg dry weight sponge was obtained after drying. This finding indicates that under our mariculture methods and harvesting strategy, the yield of H. simulans can reach approximately 900 kg dry weight biomass per hectare per year. This research provides valuable data for large-scale sponge production and new methods for the industrial development of sponge products.

Chemical Constituents and Anti-inflammatory Properties of the Marine Sponge Haliclona cratera from Konkan, India

Chemical Constituents and Anti-inflammatory Properties of the Marine Sponge Haliclona cratera from Konkan, India

A new amide from the marine sponge Haliclona baeri

A new amide, baeriamide (1), along with nine known diketopiperazines (2-10), was isolated from the marine sponge Haliclona baeri. Their structures were identified by the means of UV, IR, MS and NMR. The absolute configuration of 1 was established by Marfey’s method and comparing the specific optical rotation with the known compound HCO-Val-Gly methyl ester. Compound 1 was derived from dehydration of formylated L-valine with γ-amino-butanoic acid methyl ester. Compounds 2-10 were isolated from the genus of Haliclona for the first time. The absolute confirmation of 7 was confirmed first by the means of single-crystal X-ray diffraction. The cytotoxic, antibacterial, antiviral and antifouling activities of these compounds were also tested. However, none of them exhibited significant bioactivities.

Densazalin, a New Cytotoxic Diazatricyclic Alkaloid from the Marine Sponge Haliclona densaspicula.

Densazalin, a polycyclic alkaloid, was isolated from the marine sponge Haliclona densaspicula collected in Korea. The complete structure of the compound was determined by spectroscopic methods, including 1D and 2D nuclear magnetic resonance techniques, high-resolution mass spectrometry, and comparison of the calculated and measured electronic circular dichroism spectra. Densazalin possesses a unique 5,11-diazatricyclo[7.3.1.02,7]tridecan-2,4,6-triene moiety, which is connected by two linear carbon chains. This compound was derived from the biogenetic precursor bis-1,3-dialkylpyridnium. Densazalin exhibited cytotoxic activity on two human tumor cell lines (AGS and HepG2) in the Cell Counting Kit-8 (CCK-8) bioassay, with IC50 values ranging from 15.5 to 18.4 μM.

Antibacterial assay of crude extracts from marine sponge Haliclona fascigera in Badi Island of Spermonde Archipelago against shrimp pathogenic bacteria

The marine Sponge Haliclona fascigera, belonging to the genus Haliclona, Family Haliclonidae, Order Haplosclerida, is a source of potentially valuable marine natural products, including diverse bioactive secondary metabolites. These bioactive secondary metabolites could be used as alternative antibiotics for managing pathogenic bacteria in aquaculture. The study aimed to determine the in vitro antibacterial activity of n-hexane, ethyl acetate, and acetone crude extracts against three shrimp pathogenic bacteria and to carry out phytochemical screening. The three crude extracts were produced from H. fascigera collected around Badi Island, Spermonde Archipelago, Indonesia. The crude extracts were screened for their activity against three shrimp pathogenic bacteria: Vibrio harveyi (M-120), Vibrio alginolyticus, and Vibrio parahaemolyticus (T-170). Antibacterial activity assays used the agar diffusion method; the paper discs were impregnated with extract concentrations of 2μg/25μL. Phytochemical screening was carried out using standard protocols to provide supporting data. N-hexane and acetone crude extracts of H. fascigera were able to inhibit Vibrio parahaemolyticus with inhibition zone diameters of 8.07mm and 7.62mm, respectively. The inhibition zone of Ciprofloxacin (positive control) was 10.45 mm. The phytochemical analysis indicated that steroid, terpenoid, and alkaloid compounds were present in H. fascigera. Further studies are needed to reveal the compounds causing the observed antimicrobial effect.

Isoquinolinequinone Derivatives from a Marine Sponge (Haliclona sp.) Regulate Inflammation in In Vitro System of Intestine

Using bio-guided fractionation and based on the inhibitory activities of nitric oxide (NO) and prostaglandin E2 (PGE2), eight isoquinolinequinone derivatives (1–8) were isolated from the marine sponge Haliclona sp. Among these, methyl O-demethylrenierate (1) is a noble ester, whereas compounds 2 and 3 are new O-demethyl derivatives of known isoquinolinequinones. Compound 8 was assigned as a new 21-dehydroxyrenieramycin F. Anti-inflammatory activities of the isolated compounds were tested in a co-culture system of human epithelial Caco-2 and THP-1 macrophages. The isolated derivatives showed variable activities. O-demethyl renierone (5) showed the highest activity, while 3 and 7 showed moderate activities. These bioactive isoquinolinequinones inhibited lipopolysaccharide and interferon gamma-induced production of NO and PGE2. Expression of inducible nitric oxide synthase, cyclooxygenase-2, and the phosphorylation of MAPKs were down-regulated in response to the inhibition of NF-κB nuclear translocation. In addition, nuclear translocation was markedly promoted with a subsequent increase in the expression of HO-1. Structure-activity relationship studies showed that the hydroxyl group in 3 and 5, and the N-formyl group in 7 may be key functional groups responsible for their anti-inflammatory activities. These findings suggest the potential use of Haliclona sp. and its metabolites as pharmaceuticals treating inflammation-related diseases including inflammatory bowel disease.

Inhibition of biofilm formation and quorum sensing mediated virulence in Pseudomonas aeruginosa by marine sponge symbiont Brevibacterium casei strain Alu 1

The alternative antimicrobial strategies that mitigate the threat of antibiotic resistance is the quorum-sensing inhibition (QSI) mechanism, which targets autoinducer dependent virulence gene expression in bacterial pathogens. N-acyl homoserine lactone (AHL) acts as a key regulator in the production of virulence factors and biofilm formation in Pseudomonas aeruginosa PAO1 and violacein pigment production in Chromobacterium violaceum ATCC 12472. In the present study, the marine sponge Haliclona fibulata symbiont Brevibacterium casei strain Alu 1 showed potential QSI activity in a concentration-dependent manner (0.5–2% v/v) against the N-acyl homoserine lactone (AHL)-mediated violacein production in C. violaceum (75–95%), and biofilm formation (53–96%), protease (27–82%), pyocyanin (82–95%) and pyoverdin (29–38%) productions in P. aeruginosa. Further, the microscopic analyses validated the antibiofilm activity of the cell-free culture supernatant (CFCS) of B. casei against P. aeruginosa. Subsequently, the biofilm and pyoverdin inhibitory efficacy of the ethyl acetate extract of B. casei CFCS was assessed against P. aeruginosa. Further, the gas chromatography–mass spectrometry (GC-MS) analysis revealed the presence of variety of components in which diethyl phthalate was found to be a major active component. This phthalate ester, known as diethyl ester of phthalic acid, could act as a potential therapeutic agent for preventing bacterial biofilm and virulence associated infectious diseases.

Formal Synthesis of (-)-Haliclonin A: Stereoselective Construction of an Azabicyclo[3.3.1]nonane Ring System by a Tandem Radical Reaction.

A formal synthesis of (-)-haliclonin A, isolated from the marine sponge Haliclona sp. in Korea, is described. The key feature of the synthesis includes the highly stereoselective tandem radical reaction to construct the azabicyclo[3.3.1]nonane core and the enantioselective formation of an all-carbon quaternary center via the Pd-mediated deracemization.

The Bacterial Microbiome of the Long-Term Aquarium Cultured High-Microbial Abundance Sponge Haliclona cnidata – Sustained Bioactivity Despite Community Shifts Under Detrimental Conditions

Marine sponges host highly diverse but specific bacterial communities that provide essential functions for the sponge holobiont, including antimicrobial defense. Here, we characterized the bacterial microbiome of the marine sponge Haliclona cnidata that has been in culture in an artificial marine aquarium system. We tested the hypotheses (1) that the long-term aquarium cultured sponge Haliclona cnidata is tightly associated with typical sponge bacterial microbiota and (2) that the symbiotic Bacteria sustain bioactivity under harmful environmental conditions to facilitate holobiont survival by preventing pathogen invasion. Microscopic and phylogenetic analyses of the bacterial microbiota revealed that H. cnidata represents a high microbial abundance (HMA) sponge with a temporally stable bacterial community that significantly shifts with changing aquaria conditions. A four week incubation experiment was performed in small closed aquaria systems with antibiotic and/or light exclusion treatments to reduce the total bacterial and photosynthetically active sponge-associated microbiota to a treatment specific resilient community. While the holobiont was severely affected by the experimental treatment (i.e. fading of the sponge, reduced bacterial abundance, shifted bacterial community composition), the biological defense and bacterial community interactions (i.e. quorum sensing activity) remained intact. 16S rRNA gene amplicon sequencing revealed a resilient community of 105 bacterial taxa, which remained in the treated sponges. These 105 taxa accounted for a relative abundance of 72-83% of the bacterial sponge microbiota of non-treated sponge fragments that have been cultured under the same conditions. We conclude that a sponge-specific resilient community stays biologically active under harmful environmental conditions, facilitating the resilience of the holobiont. In H. cnidata, bacteria were located in bacteriocytes, which may have contributed to the observed phenomenon.

Topical Application of Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells in Combination with Sponge Spicules for Treatment of Photoaging.

PurposeThe topical application of exosomes secreted by mesenchymal stem cells (MSC-Exos) on the skin is a very new and interesting topic in the medical field. In this study, we aimed to investigate whether marine sponge Haliclona sp. spicules (SHSs) could effectively enhance the skin delivery of human umbilical cord-derived MSC-Exos (hucMSC-Exos), and further evaluate the topical application of hucMSC-Exos combined with SHSs in rejuvenating photoaged mouse skin.Materials and MethodsSHSs were isolated from the explants of sponge Haliclona sp. with our proprietary method, and hucMSC-Exos were prepared from the conditioned medium of hucMSCs using ultracentrifugation. The effects of SHSs on the skin penetration of fluorescently labeled hucMSC-Exos were determined using confocal microscopy in vitro (porcine skin) and in vivo (mouse skin). The therapeutic effects of hucMSC-Exos coupled with SHSs against UV-induced photoaging in mice were assessed by using microwrinkles analysis, pathohistological examination and real-time RT-PCR. We also tested the skin irritation caused by the combination of hucMSC-Exos and SHSs in guinea pigs.ResultsIn vitro results showed that hucMSC-Exos could not readily penetrate through porcine skin by themselves. However, SHSs increased the skin absorption of exosomes by a factor of 5.87 through creating microchannels. Similar penetration enhancement of hucMSC-Exos was observed after SHSs treatment in mice. The combined use of hucMSC-Exos and SHSs showed significant anti-photoaging effects in mice, including reducing microwrinkles, alleviating histopathological changes, and promoting the expression of extracellular matrix constituents, whereas hucMSC-Exos alone produced considerably weaker effects. Skin irritation test showed that the combination of hucMSC-Exos and SHSs caused slight irritation, and the skin recovered shortly.ConclusionSHSs provide a safe and effective way to enhance the skin delivery of MSC-Exos. Moreover, the combination of MSC-Exos and SHSs may be of much use in the treatment of photoaging.

X-ray Crystallography and Unexpected Chiroptical Properties Reassign the Configuration of Haliclonadiamine.

Haliclonadiamine and papuamine are bis-indane marine natural products isolated from the marine sponge Haliclona sp. Their relative structures were previously reported to differ by inversion at only one of their eight shared stereocenters. Here X-ray crystallography shows the opposite to be true: papuamine has a 1R,3S,8R,9S,14S,15R,20S,22R configuration, while haliclonadiamine has a 1S,3R,8S,9R,14R,15S,20R,22R configuration. Paradoxically the ECD of each structure displays a negative Cotton effect. X-ray crystallography reveals the two structures adopt similar conformations of their 13-membered macrocyclic core that comprises a configurationally relevant diene. B97x-D/Def2-TZVPP-(MeOH)-calculated ECD supports the diene configuration with the macrocycle dominating the ECD Cotton effect for haliclonadiamine and papuamine. Additional crystallographic and chiroptical analyses of three sponge samples from geographically distant locations indicate this pair of natural products always exists as a configurationally related couple. The co-discovery of a biosynthetic precursor, halichondriamine C, present in these same Haliclona samples must be considered when discussing any biosynthetic pathway. Taken together, this work justifies a reassignment of haliclonadiamine's structure and opens the question of how this complex stereochemical relationship between haliclonadiamine and palauamine arises biosynthetically.

The manzamine alkaloids.

The manzamine alkaloids are absolutely one of the most fascinating marine natural products. The representative manzamine alkaloids, manzamines A-C, were isolated from a marine sponge Haliclona sp. collected off Cape Manzamo, Okinawa, Japan. The manzamine alkaloids are a unique class of alkaloids possessing a characteristic heterocyclic system, and exhibit a diverse range of bioactivities including cytotoxicity, antimicrobial activity, antimalarial activity, antiviral and antiinflammatory activities, antiinsecticidal activity, and proteasome inhibitory activity. About 100 manzamine alkaloids have been isolated from more than 16 species of marine sponges belonging to 5 families. The unusual ring systems, an intriguing suggested biogenetic pathway, and promising biological activities of manzamine alkaloids have attracted great interest as challenging targets for the total synthesis. This review is the continuation of the previous review published in volume 60 of The Alkaloids and covers isolation, structure elucidation, biosynthesis and biogenesis, chemical synthesis, and biological activity of manzamine alkaloids reported from 2003 to 2018.

Synthesis and antiproliferative effect of the proposed stereoisomer of the marine sponge metabolite halisphingosine A.

The first synthesis of the proposed natural stereoisomer of halisphingosine A, a metabolite of the marine sponge Haliclona tubifera, was accomplished in 11 steps including an enantioselective Henry reaction, a Weinreb amide - acetylide coupling, and stereoselective reductions of the resulting ynone to afford the R,Z-configured allyl alcohol moiety. The synthetic product differed from the natural isolate in some 13C-NMR data. It showed antiproliferative activity at clinically relevant concentrations against six tumour cell lines including such lacking functional tumor suppressor gene p53.

High Altitude Hypoxia Alters the Iron/metalloproteins and NO Metabolites in Fetal and Maternal Sheep Blood and Aorta

The manzamine alkaloids are absolutely one of the most fascinating marine natural products. The representative manzamine alkaloids, manzamines A–C, were isolated from a marine sponge Haliclona sp. collected off Cape Manzamo, Okinawa, Japan. The manzamine alkaloids are a unique class of alkaloids possessing a characteristic heterocyclic system, and exhibit a diverse range of bioactivities including cytotoxicity, antimicrobial activity, antimalarial activity, antiviral and antiinflammatory activities, antiinsecticidal activity, and proteasome inhibitory activity. About 100 manzamine alkaloids have been isolated from more than 16 species of marine sponges belonging to 5 families. The unusual ring systems, an intriguing suggested biogenetic pathway, and promising biological activities of manzamine alkaloids have attracted great interest as challenging targets for the total synthesis. This review is the continuation of the previous review published in volume 60 of The Alkaloids and covers isolation, structure elucidation, biosynthesis and biogenesis, chemical synthesis, and biological activity of manzamine alkaloids reported from 2003 to 2018.