Spider Venom Research Articles

Animal Venom Peptides as a Treasure Trove for New Therapeutics Against Neurodegenerative Disorders.

Neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, and cerebral ischemic stroke, impose enormous socio-economic burdens on both patients and health-care systems. However, drugs targeting these diseases remain unsatisfactory, and hence there is an urgent need for the development of novel and potent drug candidates. Animal toxins exhibit rich diversity in both proteins and peptides, which play vital roles in biomedical drug development. As a molecular tool, animal toxin peptides have not only helped clarify many critical physiological processes but also led to the discovery of novel drugs and clinical therapeutics. Recently, toxin peptides identified from venomous animals, e.g. exenatide, ziconotide, Hi1a, and PcTx1 from spider venom, have been shown to block specific ion channels, alleviate inflammation, decrease protein aggregates, regulate glutamate and neurotransmitter levels, and increase neuroprotective factors. Thus, components of venom hold considerable capacity as drug candidates for the alleviation or reduction of neurodegeneration. This review highlights studies evaluating different animal toxins, especially peptides, as promising therapeutic tools for the treatment of different neurodegenerative diseases and disorders.

An Antiviral Peptide from Alopecosa nagpag Spider Targets NS2B-NS3 Protease of Flaviviruses.

Flaviviruses are single-stranded RNA viruses predominantly transmitted by the widely distributed Aedes mosquitoes in nature. As important human pathogens, the geographic reach of Flaviviruses and their threats to public health are increasing, but there is currently no approved specific drug for treatment. In recent years, the development of peptide antivirals has gained much attention. Natural host defense peptides which uniquely evolved to protect the hosts have been shown to have antiviral properties. In this study, we firstly collected the venom of the Alopecosa nagpag spider from Shangri-La County, Yunnan Province. A defense peptide named Av-LCTX-An1a (Antiviral-Lycotoxin-An1a) was identified from the spider venom, and its anti-dengue serotype-2 virus (DENV2) activity was verified in vitro. Moreover, a real-time fluorescence-based protease inhibition assay showed that An1a functions as a DENV2 NS2B–NS3 protease inhibitor. Furthermore, we also found that An1a restricts zika virus (ZIKV) infection by inhibiting the ZIKV NS2B–NS3 protease. Together, our findings not only demonstrate that An1a might be a candidate for anti-flavivirus drug but also indicate that spider venom is a potential resource library rich in antiviral precursor molecules.

Use of Spider Venom and Botanicals as Acetylcholinesterase Inhibitors Against Musca domestica (L.) (Diptera: Muscidae) and Aedes (Stegomyia) Aegypti L. (Dipetra: Culicidae)

The aim of the study was to determine insecticidal potential of three plant extracts (Calatropis procera, Eucalyptus globulus and Mentha arvensis) and crude venom of an orb-web spider, Neoscona theisi, in the laboratory using Musca domestica and Aedes aegypti as model insects. Laboratory bioassays were conducted against three doses of plants extracts (0.2 mg/ml, 0.4 mg/ml and 0.6 mg/ml) and spider venom (10 µg/ml, 20 µg/ml and 30 µg/ml). Ellman et al.'s (1961) method was followed to estimate the activity of acetylcholinesterase of insects. Calatropis procera was found to be more effective in killing the insects. The mortality rate was increased with an increase in plant extract or venom dose. Calatropis procera inhibited acetylcholinesterase more proficiently as compared to the M. arvensis and E. globulus extracts. Crude spider venom also significantly inhibited the estimated activity of acetylcholinesterase in M. domestica and Ae. aegypti. It is concluded from the study that both plant extracts and spider venom inhibited the activity of acetylcholinesterase in M. domestica and Ae. aegypti but C. procera is more efficient in killing the insects.

Determining the Structures of the Snake and Spider Toxins by X-Rays.

Snake and spider venom is a complex mixture that contains proteins, peptides, and small organic and inorganic compounds. In contrast to spider venom, snake venom proteins are well known both functionally and structurally. This work describes methods for purification and crystallization of snake and spider venom toxins and their three-dimensional structure determination by X-ray crystallography.

Snake- and Spider-Venom-Derived Toxins as Lead Compounds for Drug Development.

Snake and spider venoms have been developed by nature as a defense mechanism against predators or to immobilize their prey by blocking the cardiovascular, respiratory, and/or nervous systems. Consequently, predators are deterred from approaching their prey by painful sensations. At a molecular level, the targeted physiological systems are blocked or stimulated by peptide toxins which, once injected into the body, modulate, though not exclusively, important cell membrane ion channels and receptors. Millions of years of constant evolution have led to the evolvement of complex venom libraries of optimized protein toxins, making them more potent, more selective, resistant to proteases, less immunogenic, and improved in terms of pharmacokinetic (PK) properties. The resulting advantage is that they induce long-term and potent pharmacodynamic (PD) effects toward unique molecular targets of therapeutic importance such as coagulation cascade proteins, receptors, and ionic channels. This optimization process has been enabled by the diversification of peptide sequences (mainly by gene duplication) and an upscaling of the complexity of toxin peptide scaffold structures, through implementation of multiple disulfide bridges and sequence-active motif diversification, leading to a wide diversity of chemical structures. This combination of pharmaceutical properties has made venom toxins valuable both as pharmacological tools and as leads for drug development. These highly tunable molecules can be tailored to achieve desirable biocompatibility and biodegradability with simultaneously selective and potent therapeutic effects. This brief overview provides basic definitions, rules, and methodologies and describes successful examples of a few drugs developed from snake toxins that are currently used in the clinic for therapy of several diseases as well as new molecular entities in clinical development based on spider-venom-derived peptide toxins.

MALDI-TOF Mass Spectrometric Profiling of Spider Venoms.

Fingerprinting by means of matrix-assisted laser desorption-ionizationtime-of-flight mass spectrometry (MALDI-TOF MS) represents a tool for rapidly detecting proteinaceous compounds from spider venoms. Here we describe an optimized protocol and discuss methodological details with the aim of providing a platform for obtaining the most informative and reproducible mass spectral data.

Online, Anytime.

When Pediatrics in Review was first published 40 years ago, I was in diapers, and Visual Diagnosis was still 2 decades from conception. At its 20-year mark, I was unaware that Pediatrics in Review was beginning to publish content online. I was keenly aware of the shifting tides from traditional media to online information; after all, I was starting a career in multimedia production. However, I was still several years from discovering my second calling in pediatric medicine, so I was not yet a Pediatrics in Review reader.When I finally discovered Pediatrics in Review, it was the online version that worked best for my learning style. It has always been difficult for me to focus on reading textbooks, and the Pediatrics in Review search box solved that problem. I found that I could quickly access content relevant to my practice on demand, which made the topics come alive and allowed me to apply the knowledge to real patients.For Pediatrics in Review, the birth of online publishing opened opportunities to create additional material, and Visual Diagnosis was born in March of 2000. It was initially printed, but the format of a presenting image followed by the diagnosis and discussion allowed for a smooth transition to online-only publication. Dr Joe Zenel was the inaugural editor of this section and wrote the first article, titled “An Infant Who Has Fever and Rash.” Dr Zenel built the foundations of Visual Diagnosis and eventually became editor-in-chief. It is an honor for me to learn from him and to continue his legacy with the Visual Diagnosis feature.Dr Zenel concluded the first Visual Diagnosis article with this question: “…will the clinician-in-training recognize measles in the patient presenting with fever, cough, coryza, conjunctivitis, and a red maculopapular rash?” Given the fact that measles was eradicated from the United States in that same year, it was a valid question.Today, pocket-sized computers and excessive media hype can induce irrational fears in anxious parents. I suspect that many of you have received photographs via text message from parents asking if something looks like measles, a spider bite, or fungus. Luckily, for today’s clinicians and clinicians-in-training, there is a vast library of Visual Diagnosis cases available to access when you find yourself confronted with an unusual fever and rash. Pediatrics in Review will continue to move forward, adding to this library and offering expert content to pediatricians for many generations to come.

Maximum tolerant dose and analgesic activity of PT1 peptide

Introduction: The article presents the results of the study of the maximum tolerant dose (MTD) and the analgesic activity of peptide PT1 isolated from Alopecosa marikovskyi spider venom. PT1 is the first compound of polypeptide nature, capable of exerting a selective modulating effect on purinergic P2X3 receptors.
 Materials and methods: The study was conducted on 174 ICR mice. The analgesic activity of the peptide was evaluated in a thermal hypersensitivity test triggered by CFA and in a model of chemical irritation.
 Results and discussion: The determined MTD for the peptide PT1 when administered intravenously provides evidence to attribute it to low-toxic compounds. The maximum analgesic activity of PT1 using the biomodel of hypersensitivity induced by CFA when tested 15 minutes after the administration was recorded at doses of 0.1 and 0.5 mg/kg. In the visceral pain test, the maximum analgesic activity 15 minutes after the administration of the chemical stimulus was observed at a dose of 0.01 mg/kg.
 Conclusions: According to the results of testing peptide PT1, it is shown that it belongs to low-toxic compounds, has a pronounced analgesic activity in a wide range of doses of 0.0001–10 mg/kg.

Prey acceptance and conditional foraging behavior in the cribellate-web spider Titanoeca quadriguttata (Araneae: Titanoecidae)

Spiders employ a wide range of foraging strategies and some are known to use conditional foraging behavior, which is tailored to the type of prey attacked. Little is known about the prey capture behavior of cribellate titanoecid spiders. Here, we investigated the fundamental trophic niche and predatory behavior of a cribellate-web spider, Titanoeca quadriguttata (Hahn, 1833). Juvenile spiders were collected under stones in a quarry and used in the experiment. We offered each individual nine prey types (spider, springtail, termite, cricket, cockroach, beetle, fly, moth, and ant) and observed the frequencies of capture and consumption, and the foraging behavior. We found that T. quadriguttata accepted many prey types, and thus, that its fundamental trophic niche was wide. The hunting behavior differed among prey types, depending on the size and potential danger of the prey. Two capture behaviors were used to overcome prey on the web. For dangerous and some large prey, such as spiders and crickets, the spider bit its victim and immediately retreated, whereas for innocuous and small prey, such as springtails and flies, the spider bit its victim and held it in the chelicerae. Attacks were administered to different body parts of prey. For example, ants were always bitten on the head, and crickets were bitten on the legs. We conclude that T. quadriguttata is an euryphagous generalist predator which uses conditional prey capture behavior to catch a variety of prey types.

The bolster technique utilising small intestinal submucosa wound matrix: A novel approach to wound treatment.

Managing acute wounds with soft tissue loss can be very challenging for both patients and physicians. Successful wound healing depends on several factors including exudate control, prevention of infection, and moisture balance. In this case series, we describe a novel combination treatment method utilising small intestinal submucosa wound matrix (SISWM) with the bolster technique as a way of assisting the integration of collagen‐based wound treatment products into the base of complex wounds with the intent of restoring a dysfunctional extracellular matrix. In case 1, a 44‐year‐old female presented with an acute wound resulting from a spider bite to the posterior aspect of the right knee. In case 2, a 12‐year‐old male sustained multiple injuries to his right foot from an all‐terrain vehicle accident. In case 3, an 80‐year‐old female on anticoagulants sustained an avulsion injury to her left lower leg. In case 4, a 41‐year‐old female sustained a severe complex avulsion injury to the dorsal left forearm sustained from a dog bite. All patients were successfully treated with SISWM and the bolster technique, and their wounds healed completely within 6 weeks. The bolster technique, when combined with an SISWM, is a novel method designed to enable the SISWM to impart its wound healing properties to these complex traumatic wounds. This case series presents treating clinicians with a different treatment methodology to assist the patient in achieving a successful outcome.

Diversity of astacin-like metalloproteases identified by transcriptomic analysis in Peruvian Loxosceles laeta spider venom and in vitro activity characterization

Loxosceles spiders are found in almost all countries of South America. In Peru, Loxosceles laeta species is the main responsible for the accidents caused by poisonous animals, being known as “killer spiders”, due to the large number of fatal accidents observed. Astacin-like metalloproteases, named LALPs (Loxosceles astacin-like metalloproteases) are highly expressed in Loxosceles spiders venom gland. These proteases may be involved in hemorrhage and venom spreading, being relevant to the envenoming proccess. Thus, the aim of this work was to analyze Peruvian L. laeta venom gland transcripts using bioinformatics tools, focusing on LALPs. A cDNA library from Peruvian L. laeta venom glands was constructed and sequenced by MiSeq (Illumina) sequencer. After assembly, the resulting sequences were annotated, seeking out for similarity with previously described LALPs. Nine possible LALPs isoforms from Peruvian L. laeta venom were identified and the results were validated by in silico and in vitro experiments. This study contributes to a better understanding of the molecular diversity of Loxosceles venom and provide insights about the action of LALPs.

Evaluation of the Spider (Phlogiellus genus) Phlotoxin 1 and Synthetic Variants as Antinociceptive Drug Candidates.

Over the two last decades, venom toxins have been explored as alternatives to opioids to treat chronic debilitating pain. At present, approximately 20 potential analgesic toxins, mainly from spider venoms, are known to inhibit with high affinity the NaV1.7 subtype of voltage-gated sodium (NaV) channels, the most promising genetically validated antinociceptive target identified so far. The present study aimed to consolidate the development of phlotoxin 1 (PhlTx1), a 34-amino acid and 3-disulfide bridge peptide of a Phlogiellus genus spider, as an antinociceptive agent by improving its affinity and selectivity for the human (h) NaV1.7 subtype. The synthetic homologue of PhlTx1 was generated and equilibrated between two conformers on reverse-phase liquid chromatography and exhibited potent analgesic effects in a mouse model of NaV1.7-mediated pain. The effects of PhlTx1 and 8 successfully synthetized alanine-substituted variants were studied (by automated whole-cell patch-clamp electrophysiology) on cell lines stably overexpressing hNaV subtypes, as well as two cardiac targets, the hCaV1.2 and hKV11.1 subtypes of voltage-gated calcium (CaV) and potassium (KV) channels, respectively. PhlTx1 and D7A-PhlTx1 were shown to inhibit hNaV1.1–1.3 and 1.5–1.7 subtypes at hundred nanomolar concentrations, while their affinities for hNaV1.4 and 1.8, hCaV1.2 and hKV11.1 subtypes were over micromolar concentrations. Despite similar analgesic effects in the mouse model of NaV1.7-mediated pain and selectivity profiles, the affinity of D7A-PhlTx1 for the NaV1.7 subtype was at least five times higher than that of the wild-type peptide. Computational modelling was performed to deduce the 3D-structure of PhlTx1 and to suggest the amino acids involved in the efficiency of the molecule. In conclusion, the present structure–activity relationship study of PhlTx1 results in a low improved affinity of the molecule for the NaV1.7 subtype, but without any marked change in the molecule selectivity against the other studied ion channel subtypes. Further experiments are therefore necessary before considering the development of PhlTx1 or synthetic variants as antinociceptive drug candidates.

Demonstrating the potential of a novel spider venom-based biopesticide for target-specific control of the small hive beetle, a serious pest of the European honeybee

The parasitic small hive beetle (Aethina tumida) feeds on pollen, honey and brood of the European honey bee (Apis mellifera); establishment in North America and Australia has resulted in severe economic damage to the apiculture industry. We report potential for the “in-hive” use of a novel biopesticide that is toxic to this invasive beetle pest but harmless to honeybees. Constructs encoding the spider venom neurotoxin ω-hexatoxin-Hv1a (Hv1a) linked to the N- or C-terminus of snowdrop lectin (GNA) were used to produce recombinant Hv1a/GNA and GNA/Hv1a fusion proteins. Both were similarly toxic to beetles by injection (respective LD50s 1.5 and 0.9 nmoles/g larvae), whereas no effects on adult honeybee survival were observed at injection doses of > 200 nmoles/g insect. When fed to A. tumida larvae, GNA/Hv1a was significantly more effective than Hv1a/GNA (LC50s of 0.52 and 1.14 mg/ml diet, respectively), whereas both proteins were similarly toxic to adults. Results suggested that the reduced efficacy of Hv1a/GNA against larvae was attributable to differences in the susceptibility of the fusion proteins to cleavage by gut serine proteases. In laboratory assays, A. tumida larval survival was significantly reduced when brood, inoculated with eggs, was treated with GNA/Hv1a.

Protective response against toxic effects of Loxosceles spider venoms using recombinant mutated phospholipases-D as antigens.

Protective response against toxic effects of Loxosceles spider venoms using recombinant mutated phospholipases-D as antigens.

Latrophilins: A Neuro-Centric View of an Evolutionary Conserved Adhesion G Protein-Coupled Receptor Subfamily.

The adhesion G protein-coupled receptors latrophilins have been in the limelight for more than 20 years since their discovery as calcium-independent receptors for α-latrotoxin, a spider venom toxin with potent activity directed at neurotransmitter release from a variety of synapse types. Latrophilins are highly expressed in the nervous system. Although a substantial amount of studies has been conducted to describe the role of latrophilins in the toxin-mediated action, the recent identification of endogenous ligands for these receptors helped confirm their function as mediators of adhesion events. Here we hypothesize a role for latrophilins in inter-neuronal contacts and the formation of neuronal networks and we review the most recent information on their role in neurons. We explore molecular, cellular and behavioral aspects related to latrophilin adhesion function in mice, zebrafish, Drosophila melanogaster and Caenorhabditis elegans, in physiological and pathophysiological conditions, including autism spectrum, bipolar, attention deficit and hyperactivity and substance use disorders.

When spider and snake get along: Fusion of a snake disintegrin with a spider phospholipase D to explore their synergistic effects on a tumor cell

Venoms of spiders and snakes contain toxins extremely active and, thus, provide a natural source for the development of new biotechnological tools. Among the diversity of toxins present in the venom of spiders from genus Loxosceles, the phospholipases D (PLDs) show high hydrolytic activity upon lysophosphatidylcholine (LPC) and sphingomyelin (SM), generating bioactive phospholipids such as cyclic phosphatidic acid (cPA). Since this mediator has been shown to play a major role in complex signaling pathways, including inhibition of tumor cells, the PLDs may hold the key to learn how toxins could be used for therapeutic purposes. However, the strong platelet aggregation of PLDs and their lack of selectivity impose a major limitation. On the other hand, disintegrins present in the venoms of Viperidae snakes are a potent inhibitor of platelet aggregation and possess high affinity and specificity to molecules called integrins that are highly expressed in some tumor cells, such as murine melanoma B16F10. Therefore, disintegrins might be suitable molecules to carry the PLDs to the malignant cells, so both toxins may work synergistically to eliminate these cells. Thus, in this work, a recombinant PLD from Loxosceles gaucho spider was recombinantly fused to a disintegrin from Echis carinatus snake to form a hybrid toxin called Rechistatin. This recombinant toxin was successfully expressed in bacteria, showed binding activity in B16F10 murine melanoma cells and exerted a synergistic cytotoxicity effect on these cells. Therefore, the approach presented in this work may represent a new strategy to explore new potential applications for spider PLDs.

Along came a spider: Medicine’s most famous spider eponyms

Spiders have long been admired for the beauty of their webs. They are celebrated in popular culture as well as in medical eponyms. This contribution provides the historical background for three spider-related eponyms: nevus araneus (spider angioma), arachnodactyly, and the arachnoid mater. Nevus araneus was first named and described by Sir Erasmus Wilson in 1842. Arachnodactyly was described in 1896 by Antoine Marfan using the term pattes d’araignée, which means spider legs. In 1902, Emile Charles Achard proposed the term arachnodactyly for this clinical finding. The arachnoid mater had been named in 1699 by Frederik Ruysch. The clinical management of spider bites from the only two dangerous venomous spiders within the United States, the black widow spider (Latrodectus mactans) and the brown recluse spider (Loxosceles reclusa), is reviewed.

Brown Spider (Loxosceles) Venom Toxins as Potential Biotools for the Development of Novel Therapeutics.

Brown spider envenomation results in dermonecrosis with gravitational spreading characterized by a marked inflammatory reaction and with lower prevalence of systemic manifestations such as renal failure and hematological disturbances. Several toxins make up the venom of these species, and they are mainly peptides and proteins ranging from 5–40 kDa. The venoms have three major families of toxins: phospholipases-D, astacin-like metalloproteases, and the inhibitor cystine knot (ICK) peptides. Serine proteases, serpins, hyaluronidases, venom allergens, and a translationally controlled tumor protein (TCTP) are also present. Toxins hold essential biological properties that enable interactions with a range of distinct molecular targets. Therefore, the application of toxins as research tools and clinical products motivates repurposing their uses of interest. This review aims to discuss possibilities for brown spider venom toxins as putative models for designing molecules likely for therapeutics based on the status quo of brown spider venoms. Herein, we explore new possibilities for the venom components in the context of their biochemical and biological features, likewise their cellular targets, three-dimensional structures, and mechanisms of action.

Vestaron gets cash for biopesticides

Kalamazoo, Michigan–based Vestaron has closed a $40 million series B financing round led by Novo Holdings, parent company of Novo Nordisk and Novozymes. Vestaron develops biopesticides based on peptides found in spider venom. The start-up says they could prove to be environmentally friendly alternatives to synthetic crop protection chemicals. The company thus far has two products: one for greenhouse pests such as thrips, whiteflies, aphids, and mites and another for lepidopterans such as moths.

Tying pest insects in knots: the deployment of spider-venom-derived knottins as bioinsecticides.

Spider venoms are complex chemical arsenals that contain a rich variety of insecticidal toxins. However, the major toxin class in many spider venoms is disulfide-rich peptides known as knottins. The knotted three-dimensional fold of these mini-proteins provides them with exceptional chemical and thermal stability as well as resistance to proteases. In contrast with other bioinsecticides, which are often slow-acting, spider knottins are fast-acting neurotoxins. In addition to being potently insecticidal, some knottins have exceptional taxonomic selectivity, being lethal to key agricultural pests but innocuous to vertebrates and beneficial insects such as bees. The intrinsic oral activity of these peptides, combined with the ability of aerosolized knottins to penetrate insect spiracles, has enabled them to be developed commercially as eco-friendly bioinsecticides. Moreover, it has been demonstrated that spider-knottin transgenes can be used to engineer faster-acting entomopathogens and insect-resistant crops. © 2019 Society of Chemical Industry.