A Transcriptomic Approach to the Recruitment of Venom Proteins in a Marine Annelid

CEECIND/02699/2017The growing number of known venomous marine invertebrates indicates that chemical warfare plays an important role in adapting to diversified ecological niches, even though it remains unclear how toxins fit into the evolutionary history of these animals. Our case study, the Polychaeta Eulalia sp., is an intertidal predator that secretes toxins. Whole-transcriptome sequencing revealed proteinaceous toxins secreted by cells in the proboscis and delivered by mucus. Toxins and accompanying enzymes promote permeabilization, coagulation impairment and the blocking of the neuromuscular activity of prey upon which the worm feeds by sucking pieces of live flesh. The main neurotoxins ("phyllotoxins") were found to be cysteine-rich proteins, a class of substances ubiquitous among venomous animals. Some toxins were phylogenetically related to Polychaeta, Mollusca or more ancient groups, such as Cnidaria. Some toxins may have evolved from non-toxin homologs that were recruited without the reduction in molecular mass and increased specificity of other invertebrate toxins. By analyzing the phylogeny of toxin mixtures, we show that Polychaeta is uniquely positioned in the evolution of animal venoms. Indeed, the phylogenetic models of mixed or individual toxins do not follow the expected eumetazoan tree-of-life and highlight that the recruitment of gene products for a role in venom systems is complex.publishersversionpublishe

Comprehensive review on the interaction between natural compounds and brain receptors: Benefits and toxicity

Given their therapeutic activity, natural products have been used in traditional medicines throughout the centuries. The growing interest of the scientific community in phytopharmaceuticals, and more recently in marine products, has resulted in a significant number of research efforts towards understanding their effect in the treatment of neurodegenerative diseases, such as Alzheimer's (AD), Parkinson (PD) and Huntington (HD). Several studies have shown that many of the primary and secondary metabolites of plants, marine organisms and others, have high affinities for various brain receptors and may play a crucial role in the treatment of diseases affecting the central nervous system (CNS) in mammalians. Actually, such compounds may act on the brain receptors either by agonism, antagonism, allosteric modulation or other type of activity aimed at enhancing a certain effect. The current manuscript comprehensively reviews the state of the art on the interactions between natural compounds and brain receptors. This information is of foremost importance when it is intended to investigate and develop cutting-edge drugs, more effective and with alternative mechanisms of action to the conventional drugs presently used for the treatment of neurodegenerative diseases. Thus, we reviewed the effect of 173 natural products on neurotransmitter receptors, diabetes related receptors, neurotrophic factor related receptors, immune system related receptors, oxidative stress related receptors, transcription factors regulating gene expression related receptors and blood-brain barrier receptors.The author A.R.S. is grateful to the authors C.G. and J.M.R for the scientific assistance and suggestions shared throughout the supervision of her B.Sc. final project. The author A.R.S. also acknowledges the Department of Biology (DB) and the Centro de Biologia Molecular e Ambiental [Centre of Molecular and Environmental Biology] (CBMA) from School of Sciences (EC), University of Minho (UM), Braga, Portugal, by providing all the conditions leading to the B.Sc. in Biochemistry. The corresponding author C.G. is grateful to Fundação para a Ciência e a Tecnologia [Foundation for Science and Technology, FCT I.P.] (FCT I.P.) for the FCT Investigator (IF/01332/2014/CP1255/CT0001). The author J.M.R. acknowledges the CBMA and the Instituto de Ciência e Inovação para a Biosustentabilidade [Institute of Science and Innovation for Biosustainability] (IBS) from University of Minho (UM), Braga, Portugal, where he recently carried out his activities as invited assistant researcher and professor. The author J.M.R. is also grateful to the Laboratório Associado para a Química Verde (LAQV) [Green Chemistry Laboratory] from REQUIMTE – Rede de Química e Tecnologia [REQUIMTE – Chemistry and Technology Network], as well as to the Department of Chemistry and Biochemistry (DCB) from the Faculty of Sciences from University of Porto (FCUP), Porto, Portugal, where he is currently researcher. Regarding to the author J.M.R., this work was supported by the strategic programmes UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) (Research project entitled “EcoAgriFood: Innovative green products and processes to promote Agri-Food BioEconomy”) and PTDC/SAUNUT/30448/2017 (POCI-01-0145-FEDER-030448) (Research project entitled “Poly4CD: Action of Dietary Polyphenols in Preventing Celiac Disease”) funded by Portuguese national funds through Fundação para a Ciência e Tecnologia [Foundation for Science and Technology] (FCT-I.P.)/Ministério da Ciência, Tecnologia e Ensino Superior [Ministry of Science, Technology and Higher Education] (MCTES), and Fundo Europeu de Desenvolvimento Regional [European Regional Development Fund] (FEDER), under the scope of the COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI) [COMPETE2020 – Competitiveness and Internationalization Operational Program] and by COST Action 18101 “SOURDOMICS”, supported by COST (European Cooperation in Science and Technology). C. G. and C. D.-M. are grateful for the financial support from FCT/MCTES through national funds (UID/QUI/50006/2019). C. G. and C. D.-M. would also like to thank the EU and FCT for funding through the projects: DESignBIOtecHealth - New Technologies for three Health Challenges of Modern Societies: Diabetes, Drug Abuse and Kidney Diseases (Portugal2020, Norte-01-0145-FEDER-000024) and project PTDC/OCE-ETA/30240/2017- SilverBrain - From sea to brain: Green neuroprotective extracts for nanoencapsulation and functional food production (POCI-01-0145-FEDER-030240); and to the REQUIMTE for the project “Sea_4_Brain_Food”. All the authors acknowledge the CBMA and IB-S by the financial support provided specifically for this open access publication.info:eu-repo/semantics/publishedVersio

surfing the deep blue sea for novel bioactive proteins and peptides

Funding Information: The author(s) declare financial support was received for the research, authorship, and/or publication of this article. Fundo Azul co-financed project MARVEN (FA_05_2017_007). The Portuguese Foundation for Science and Technology (FCT), I. P., funded project WormALL (PTDC/BTA-BTA/28650/2017). This work was also financed by the National Funds from FCT in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences \u2013 UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy \u2013 i4HB. FCT is also acknowledged for the grants SFRH/BD/144914/2019 to CG and 2022.11150.BD to IMC. Acknowledgments Publisher Copyright: Copyright © 2024 Moutinho Cabral, Gonçalves, Grosso and Costa.The vast biological and biochemical diversity of the global ocean is the driver behind marine bioprospecting for novel bioproducts. As Marine Biotechnology is gaining momentum as one of the main pillars of the ‘Brue Growth’ revolution, the ability to screen for novel compounds of interest in species with little or no genomic resources is paramount. With this respect, proteins, which are easily metabolised, can be synthetised using convenient DNA recombinant methods and can easily be modified to better meet the needs of human society, making them prized targets. Evidently, proteins that hold natural bioactivity and specificity such as toxins and other venom components, have long captured the focus of biotechnologists, leading to the merger between environmental omics and toxinology termed as ‘venomics’. Indeed, bioactive proteins such as conopeptides, conotoxins, turripeptides and others are long deemed important subjects of research. Even though current mainstream paradigms set the focus on secondary metabolites from marine organisms, transcriptomics and proteomics approaches and their combination are rising strategies for screening for thousands of proteins and peptides in non-conventional biological models, emphasising, but not limited to, marine invertebrate animals due to their abundance, biodiversity and uncanny biochemical strategies to cope with selective pressure in literally every known marine habitat. Untargeted approaches, such as RNA-Seq - based transcriptomics and tandem mass spectrometry - based proteomics, can circumvent limitations related with absent or reduced genomic annotation. The present review will outline the main contributions of ‘omics’ and computational approaches for bioprospecting for proteinaceous marine bioactives. Despite the relatively low number of ‘omics’ studies with the main purpose of discover novel compounds, there is already important literature showcasing pipelines and approaches for revolutionising the exploration of the ocean.publishersversionpublishe

Genetic predisposition for aggressive behaviour related with dopamine and serotonin pathways : an overview

Abstract in proceedings of the Fourth International Congress of CiiEM: Health, Well-Being and Ageing in the 21st Century, held at Egas Moniz’ University Campus in Monte de Caparica, Almada, from 3–5 June 2019.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.info:eu-repo/semantics/publishedVersio

Transcriptome profiling of the posterior salivary glands of the cuttlefish Sepia officinalis from the Portuguese West coast

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The authors acknowledge \u201CFundo azul\u201D for the funding of research project MARVEN (Ref. FA_05_2017_007). This work is also financed by national funds from The Portuguese Foundation for Science and Technology (FCT), I.P., in the scope of project UIDP/04378/2020 and UIDB/04378/2020 of UCIBIO and project LA/P/0140/2020 of i4HB. FCT is also acknowledged for the grants SFRH/BD/144914/2019 to CG and 2022.11150.BD to IMC. Acknowledgments Publisher Copyright: Copyright © 2024 Gonçalves, Moutinho Cabral, Alves de Matos, Grosso and Costa.Cephalopods like octopuses and cuttlefishes are known to secrete a ‘toxic saliva’ to inject into their prey, especially crustaceans since the XIX century. However, only in the mid-XX century were the first coleoid-specific toxins successfully isolated. Motivated by the growing interest on the global ocean as an almost inexhaustible source of novel bioactive compounds, we used RNA-Seq - based transcriptomics and de novo assembly of transcriptomes to screen the posterior salivary glands of Sepia officinalis (the common cuttlefish) from the Portuguese West coast for toxins and other bioactive proteins and peptides. Supported by microanatomical analyses, the posterior salivary glands constitute indeed the ‘venom gland’ whereas the more elusive anterior salivary glands (embedded in the buccal mass) are responsible for the production of mucin-rich saliva that is effectively the vehicle that transports the toxins as the venom is injected into the prey. Indeed, the transcriptomic profiling suggests that the cuttlefish venom is complex mixture of bioactive proteins, among which neurotoxins are major players, together with enzymes whose function is to digest the extracellular matrix to facilitate diffusion of the toxins. Nonetheless, by comparing with previous RNA-Seq data obtained from S. officinalis collected from other biogeographical areas, it may be suggested that significant inter-populational variation in venom composition can occur, which may potentially increase the span of bioactives secreted by these animals. We isolated and validated the full coding sequences for three important toxins, a cysteine-rich venom protein (CRVP), a venom insulin (VIns) and a cephalotoxin (CTX). The toxins seem to be relatively conserved among coleoids but diverging from other venomous mollusks such as cone snails. Their properties as potent modulators of glucose (in the case of VIns) and as potential neurotoxins (like CRVP and CTX) can render them primer targets for drug development.publishersversionpublishe

A novel bacterium for 2-phenylethanol production

Funding Information: This work was supported by the Associated Laboratory for Green Chemistry \u2013 LAQV (UIDB/50006/2020 and UIDP/50006/2020), the Associate Laboratory Institute for Health and Bioeconomy - i4HB (LA/P/0140/2020), the Applied Molecular Biosciences Unit \u2013 UCIBIO (UIDP/04378/2020 and UIDB/04378/2020) and the national project PTDC/CTM-CTM/29869/2017 and UID/DTP/04138/2019, which are financed by national funds from FCT - Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia, I.P. (Portugal). This work also received financial support from ACINETOBACTER_POCI-01-0145-FEDER-042759. Funding Information: Ana R. Bernardino acknowledge FCT, I.P. for financial support through PhD fellowship SFRH/BD/138011/2018 and Filipa Grosso was supported by national funds through FCT in the context of the transitional norm (DL57/2016/CP1346/CT0034; ( https://doi.org/10.54499/DL57/2016/CP1346/CT0034 ). Publisher Copyright: © 2024A bacterium, Acinetobacter soli ANG344B, isolated from river water, exhibited an exceptional capacity to produce 2-phenylethanol (2-PE) using L-phenylalanine (L-Phe) as a precursor—a capability typically observed in yeasts rather than bacteria. Bioreactor experiments were conducted to evaluate the production performance, using glucose as the carbon source for cellular growth and L-Phe as the precursor for 2-PE production. Remarkably, A. soli ANG344B achieved a 2-PE concentration of 2.35 ± 0.26 g/L in just 24.5 h of cultivation, exhibiting a global volumetric productivity of 0.10 ± 0.01 g/L.h and a production yield of 0.51 ± 0.01 g2-PE/gL-Phe, a result hitherto reported only for yeasts. These findings position A. soli ANG344B as a highly promising microorganism for 2-PE production. Whole-genome sequencing of A. soli strain ANG344 revealed a genome size of 3.52 Mb with a GC content of 42.7 %. Utilizing the Rapid Annotation using Subsystem Technology (RAST) server, 3418 coding genes were predicted, including genes coding for enzymes previously associated with the metabolic pathway of 2-PE production in other microorganisms, yet unreported in Acinetobacter species. Through gene mapping, 299 subsystems were identified, exhibiting 30 % subsystem coverage. The whole genome sequence data was submitted to NCBI GeneBank with the BioProject ID PRJNA982713. These draft genome data offer significant potential for exploiting the biotechnological capabilities of A. soli strain ANG344 and for conducting further comparative genomic studies.publishersversionpublishe

The genetic susceptibility linking preterm birth and periodontal disease : a review

Abstract in proceedings of the Fourth International Congress of CiiEM: Health, Well-Being and Ageing in the 21st Century, held at Egas Moniz’ University Campus in Monte de Caparica, Almada, from 3–5 June 2019.This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.info:eu-repo/semantics/publishedVersio

Specific Antiproliferative Properties of Proteinaceous Toxin Secretions from the Marine Annelid Eulalia sp. onto Ovarian Cancer Cells

The Portuguese Foundation for Science and Technology (FCT) funded project WormALL (PTDC/BTA-BTA/28650/2017) plus the grants SFRH/BD/109462/2015 to A.P.R., and CEECIND/02699/2017 to A.R.G. The Applied Molecular Biosciences Unit-UCIBIO is financed by national funds from FCT, ref. UIDB/04378/2020. FCT, along with the European Regional Development Fund (ERDF) through the COMPETE 2020-Operational Programme for Competitiveness and Internationalisation also funded the projects: POCI-01-0145-FEDER-007440 (UID/NEU/04539/2019), POCI-01-0145-FEDER-016428 (SAICTPAC/0010/2015), POCI-01-0145-FEDER-029311 (PTDC/BTM-TEC/29311/2017), POCI-01-0145-FEDER-30943 (PTDC/MEC-PSQ/30943/2017) and PTDC/MED-NEU/27946/2017. The work was also funded by the National Mass Spectrometry Network (RNEM) under the contract POCI-01-0145-FEDER-402-022125 (ref.: ROTEIRO/0028/2013).As Yondelis joins the ranks of approved anti-cancer drugs, the benefit from exploring the oceans' biodiversity becomes clear. From marine toxins, relevant bioproducts can be obtained due to their potential to interfere with specific pathways. We explored the cytotoxicity of toxin-bearing secretions of the polychaete Eulalia onto a battery of normal and cancer human cell lines and discovered that the cocktail of proteins is more toxic towards an ovarian cancer cell line (A2780). The secretions' main proteins were identified by proteomics and transcriptomics: 14-3-3 protein, Hsp70, Rab3, Arylsulfatase B and serine protease, the latter two being known toxins. This mixture of toxins induces cell-cycle arrest at G2/M phase after 3h exposure in A2780 cells and extrinsic programmed cell death. These findings indicate that partial re-activation of the G2/M checkpoint, which is inactivated in many cancer cells, can be partly reversed by the toxic mixture. Protein-protein interaction networks partake in two cytotoxic effects: cell-cycle arrest with a link to RAB3C and RAF1; and lytic activity of arylsulfatases. The discovery of both mechanisms indicates that venomous mixtures may affect proliferating cells in a specific manner, highlighting the cocktails' potential in the fine-tuning of anti-cancer therapeutics targeting cell cycle and protein homeostasis.publishersversionpublishe

Aerocyte specification and lung adaptation to breathing is dependent on alternative splicing changes

© 2022 Fidalgo et al. This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).Adaptation to breathing is a critical step in lung function and it is crucial for organismal survival. Alveoli are the lung gas exchange units and their development, from late embryonic to early postnatal stages, requires feedbacks between multiple cell types. However, how the crosstalk between the alveolar cell types is modulated to anticipate lung adaptation to breathing is still unclear. Here, we uncovered a synchronous alternative splicing switch in multiple genes in the developing mouse lungs at the transition to birth, and we identified hnRNP A1, Cpeb4, and Elavl2/HuB as putative splicing regulators of this transition. Notably, we found that Vegfa switches from the Vegfa 164 isoform to the longer Vegfa 188 isoform exclusively in lung alveolar epithelial AT1 cells. Functional analysis revealed that VEGFA 188 (and not VEGFA 164) drives the specification of Car4-positive aerocytes, a subtype of alveolar endothelial cells specialized in gas exchanges. Our results reveal that the cell type-specific regulation of Vegfa alternative splicing just before birth modulates the epithelial-endothelial crosstalk in the developing alveoli to promote lung adaptation to breathing.This work was supported by European Research Council (ERC starting grant [679368]), the European Union (H2020-TWINN-2015 – Twinning [692322]), Fundação para a Ciência e Tecnologia (FCT) (PTDC/MED-PAT/31639/2017, and UIDP/04378/2020 of the Research Unit on Applied Molecular Biosciences - UCIBIO), and Fondation Leducq (17CVD03). CG Fonseca was supported by a PhD fellowship from the doctoral program Bioengineering: Cellular Therapies and Regenerative Medicine funded by Fundação para a Ciência e Tecnologia (FCT) (PD/BD/128375/2017). T Balboni was supported by a PhD fellowship from the doctoral program “Oncology, Hematology and Pathology - 30th Cycle” funded by University of Bologna, Italy. P Caldas was supported by a postdoctoral researcher fellowship from FCT (PTDC/MED-ONC/28660/2017). AASF Raposo was supported by FCT and Fundo Europeu de Desenvolvimento Regional (FEDER) PAC-PRECISE-LISBOA-01-0145-FEDER-016394 and by an assistant researcher contract from FCT (CEECIND/01474/2017). AR Grosso was supported by a principal investigator contract from FCT (CEECIND/02699/2017). FF Vasconcelos was supported by a postdoctoral researcher contract from FCT (CEECIND/04251/2017). CA Franco was supported by a principal investigator contract from FCT (CEECIND/02589/2018).info:eu-repo/semantics/publishedVersio

Sialyl LewisX/A and Cytokeratin Crosstalk in Triple Negative Breast Cancer

project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB. Publisher Copyright: © 2023 by the authors.Triple-negative breast cancer (TNBC) encompasses multiple entities and is generally highly aggressive and metastatic. We aimed to determine the clinical and biological relevance of Sialyl-Lewis X and A (sLeX/A)—a fucosylated glycan involved in metastasis—in TNBC. Here, we studied tissues from 50 TNBC patients, transcripts from a TNBC dataset from The Cancer Genome Atlas (TCGA) database, and a primary breast cancer cell line. All 50 TNBC tissue samples analysed expressed sLeX/A. Patients with high expression of sLeX/A had 3 years less disease-free survival than patients with lower expression. In tissue, sLeX/A negatively correlated with cytokeratins 5/6 (CK5/6, which was corroborated by the inverse correlation between fucosyltransferases and CK5/6 genes. Our observations were confirmed in vitro when inhibition of sLeX/A remarkably increased expression of CK5/6, followed by a decreased proliferation and invasion capacity. Among the reported glycoproteins bearing sLeX/A and based on the STRING tool, α6 integrin showed the highest interaction score with CK5/6. This is the first report on the sLeX/A expression in TNBC, highlighting its association with lower disease-free survival and its inverse crosstalk with CK5/6 with α6 integrin as a mediator. All in all, sLeX/A is critical for TNBC malignancy and a potential prognosis biomarker and therapeutic target.publishersversionpublishe