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"Chakir, Bouchra"
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Anti-Cancer Activity of Buthus occitanus Venom on Hepatocellular Carcinoma in 3D Cell Culture
Hepatocellular carcinoma (HCC) is the most dominant primary liver cancer, which can be caused by chronic hepatitis virus infections and other environmental factors. Resection, liver transplantation, and local ablation are only a few of the highly effective and curative procedures presently accessible. However, other complementary treatments can reduce cancer treatment side effects. In this present work, we evaluated the activity of Moroccan scorpion venom Buthus occitanus and its fractions obtained by chromatography gel filtration against HCC cells using a 3D cell culture model. The venom was fractionated by gel filtration chromatography, each fraction and the crude venom was tested on normal hepatocytes (Fa2N-4 cells). Additionally, the fractions and the crude venom were tested on MCTSs (multicellular tumor spheroids), and this latter was generated by cultivate Huh7.5 cancer cell line with WI38 cells, LX2 cells, and human endothelial cells (HUVEC). Our results indicate that Buthus occitanus venom toxin has no cytotoxic effects on normal hepatocytes. Moreover, it is reported that F3 fraction could significantly inhibit the MCTS cells. Other Protein Separation Techniques (High-performance liquid chromatography) are needed in order to identify the most active molecule.
Journal Article
Moroccan Naja haje Venom and its Peptides: In Vivo Toxicity and In Vitro Antiproliferative Effect on Hepatocellular Carcinoma HepG2 Cells
Purpose
Snake venom, despite its toxicity, presents notable therapeutic potential by specifically targeting tumor cells to inhibit their proliferation or induce their apoptosis. However, with the global rise in hepatocellular carcinoma rates, new treatments are needed that move away from surgery and toxic chemotherapy. Research has revealed that snake venoms, particularly those of cobras, contain promising molecules for the treatment of cancer.
Methods
Our study evaluates the toxicity (LD
50
) of the Moroccan cobra venom
Naja haje
as well as its fractions obtained by gel filtration fractionation. We also analyze their antiproliferative activity on HepG2 hepatocellular carcinoma cells. Cell viability was determined by MTT assay and the fractions that exhibited proliferation inhibitions of HepG2 cells were analyzed by mass spectrometry (LC-MS/MS).
Results
Our results show that the LD
50
of
Naja haje
venom varies depending on the route of administration, 0.69, 0.79 and 0.90 µg/g by intracerebroventricular, intravenous and intraperitoneal routes, respectively. On the other hand, the fractions, notably 1, 2, 3, 7 and 8, demonstrate significant antiproliferative activity on the HepG2 cells in vitro, and this activity is enhanced at a concentration of 50 µg/mL, highlighting their therapeutic potential. LC-MS/MS analysis of the active fractions revealed a complex molecular composition, including cytotoxins, neurotoxins, cardiotoxins, and a diverse array of peptides.
Conclusions
Fractions 7 and 8 from
Naja haje
venom showed significant antiproliferative effects on HepG2 cells at 10 µg/mL, enhanced at 50 µg/mL.
Journal Article
Development and Efficacy of the Antivenom Specific to Severe Envenomations in Morocco and North Africa: Advancements in Scorpion Envenomation Management
Scorpion envenomation poses a global public health issue, with an estimated 1,500,000 cases worldwide annually resulting in 2600 deaths. North Africa, particularly Morocco, experiences severe envenomations, mainly attributed to Androctonus mauretanicus and Buthus occitanus in Morocco, and Buthus occitanus and Androctonus australis hector in Algeria and Tunisia, with case numbers often underestimated. Current treatment relies mainly on symptomatic approaches, except in Morocco, where management is limited to symptomatic treatment due to controversies regarding specific treatment. In Morocco, between 30,000 and 50,000 scorpion envenomation cases are reported annually, leading to hundreds of deaths, mainly among children. Controversies among clinicians persist regarding the appropriate course of action, often limiting treatments to symptomatic measures. The absence of a specific antivenom for the venoms of the most lethal scorpions further exacerbates the situation. This study aims to address this gap by developing a monovalent antivenom against the endemic and most dangerous scorpion, Androctonus mauretanicus. The antivenom was produced by immunizing albino rabbits with a mixture of Androctonus mauretanicus venom collected from high-risk areas in Morocco. Immunizations were performed by subcutaneous injections at multiple sites near the lymphatic system, following an immunization schedule. Production control of neutralizing antibody titers was conducted through immunodiffusion. Once a sufficient antibody titer was achieved, blood collection was performed, and the recovered plasma underwent affinity chromatography. The efficacy of purified IgG was evaluated by determining the ED50 in mice, complemented by histological and immunohistochemical studies on its ability to neutralize venom-induced tissue alterations and the neutralization of toxins bound to receptors in the studied organs. The monovalent antivenom demonstrated specificity against Androctonus mauretanicus venom and effective cross-protection against the venom of the scorpions Buthus occitanus and Androctonus australis hector, highly implicated in lethal envenomations in the Maghreb. This study shows that the developed monovalent antivenom exhibits notable efficacy against local scorpions and a surprising ability to neutralize the most lethal envenomations in North Africa. These results pave the way for a new, more specific, and promising therapeutic approach to countering severe scorpion envenomations, especially in Morocco, where specific treatment is lacking.
Journal Article
C-band radar data and in situ measurements for the monitoring of wheat crops in a semi-arid area (center of Morocco)
A better understanding of the hydrological functioning of irrigated crops using remote sensing observations is of prime importance in semi-arid areas where water resources are limited. Radar observations, available at high resolution and with a high revisit time since the launch of Sentinel-1 in 2014, have shown great potential for the monitoring of the water content of the upper soil and of the canopy. In this paper, a complete set of data for radar signal analysis is shared with the scientific community for the first time to our knowledge. The data set is composed of Sentinel-1 products and in situ measurements of soil and vegetation variables collected during three agricultural seasons over drip-irrigated winter wheat in the Haouz plain in Morocco. The in situ data gather soil measurements (time series of half-hourly surface soil moisture, surface roughness and agricultural practices) and vegetation measurements collected every week/2 weeks including aboveground fresh and dry biomasses, vegetation water content based on destructive measurements, the cover fraction, the leaf area index, and plant height. Radar data are the backscattering coefficient and the interferometric coherence derived from Sentinel-1 GRDH (Ground Range Detected High Resolution) and SLC (Single Look Complex) products, respectively. The normalized difference vegetation index derived from Sentinel-2 data based on Level-2A (surface reflectance and cloud mask) atmospheric-effects-corrected products is also provided. This database, which is the first of its kind made available open access, is described here comprehensively in order to help the scientific community to evaluate and to develop new or existing remote sensing algorithms for monitoring wheat canopy under semi-arid conditions. The data set is particularly relevant for the development of radar applications including surface soil moisture and vegetation variable retrieval using either physically based or empirical approaches such as machine and deep learning algorithms. The database is archived in the DataSuds repository and is freely accessible via the following DOI: https://doi.org/10.23708/8D6WQC (Ouaadi et al., 2020a).
Journal Article
Anti-Cancer Effect of Moroccan Cobra Naja haje Venom and Its Fractions against Hepatocellular Carcinoma in 3D Cell Culture
Hepatocellular carcinoma (HCC) is the most common primary liver cancer in adults, the fifth most common malignancy worldwide and the third leading cause of cancer related death. An alternative to the surgical treatments and drugs, such as sorafenib, commonly used in medicine is necessary to overcome this public health problem. In this study, we determine the anticancer effect on HCC of Moroccan cobra Naja haje venom and its fraction obtained by gel filtration chromatography against Huh7.5 cancer cell line. Cells were grown together with WI38 human fibroblast cells, LX2 human hepatic stellate cell line, and human endothelial cells (HUVEC) in MCTS (multi-cellular tumor spheroids) models. The hepatotoxicity of venom and its fractions were also evaluated using the normal hepatocytes cell line (Fa2N-4 cells). Our results showed that an anti HCC activity of Moroccan cobra Naja haje venom and, more specifically, the F7 fraction of gel filtration chromatography exhibited the greatest anti-hepatocellular carcinoma effect by decreasing the size of MCTS. This effect is associated with a low toxicity against normal hepatocytes. These results strongly suggest that the F7 fraction of Moroccan cobra Naja haje venom obtained by gel filtration chromatography possesses the ability to inhibit cancer cells proliferation. More research is needed to identify the specific molecule(s) responsible for the anticancer effect and investigate their mechanism of action.
Journal Article
Mass spectrometry‐based top‐down and bottom‐up approaches for proteomic analysis of the Moroccan Buthus occitanus scorpion venom
Buthus occitanus (B. occitanus) is one of the most dangerous scorpions in the world. Despite the involvement of B. occitanus scorpion in severe cases of envenomation in Morocco, no study has focused yet on the proteomic composition of the Moroccan B. occitanus scorpion venom. Mass spectrometry‐based proteomic techniques are commonly used in the study of scorpion venoms. The implementation of top‐down and bottom‐up approaches for proteomic analyses facilitates screening by allowing a global view of the structural aspects of such complex matrices. Here, we provide a partial overview of the venom of B. occitanus scorpion, in order to explore the diversity of its toxins and hereafter understand their effects. To this end, a combination of top‐down and bottom‐up approaches was applied using nano‐high liquid chromatography coupled to nano‐electrospray tandem mass spectrometry (nano‐LC‐ESI MS/MS). The LC‐MS results showed that B. occitanus venom contains around 200 molecular masses ranging from 1868 to 16 720 Da, the most representative of which are those between 5000 and 8000 Da. Interestingly, combined top‐down and bottom‐up LC‐MS/MS results allowed the identification of several toxins, which were mainly those acting on ion channels, including those targeting sodium (NaScTxs), potassium (KScTxs), chloride (ClScTxs), and calcium channels (CaScTx), as well as antimicrobial peptides (AMPs), amphipathic peptides, myotropic neuropeptides, and hypothetical secreted proteins. This study reveals the molecular diversity of B. occitanus scorpion venom and identifies components that may have useful pharmacological activities. Buthus occitanus scorpion possesses highly toxic venom. This article describes the first proteomic screening of the Moroccan B. occitanus venom. Our findings highlight the complexity and the toxin diversity of this venom. The most representative compounds are neurotoxins. In addition, antimicrobial peptides, amphipathic peptides, myotropic neuropeptides, and hypothetical secreted proteins were also detected in this venom.
Journal Article