The work of our group has been focused on the interaction between the genome and environmental stressors as determinant of human conditions. In pursue of this global objective, two main research areas have been defined: i) environmental genotoxicity; ii) Molecular genetics and mechanism of different human diseases. Ultimately, our research intends to provide knowledge, technologies and strategies to prevent environment-associated and genetic diseases.
ENVIRONMENTAL DETERMINANTS OF HUMAN DISEASE
Human exposure to physical, biological or chemical stressors (and their mixtures) that are present in environmental or in occupational settings may generate adverse effects in somatic cells’ genome. The field of environmental genotoxicity investigates the environmental risk factors that may impact the human genome and the consequent genomic instability that may lead to genetic diseases, with emphasis on cancer. Environmental stressor that have been recently investigated for their genotoxicity include mycotoxins and their mixtures, chemical pollutants that persist in freshwater sediments, water disinfection by-products, tobacco smoke and nanomaterials. The studies underway have been focused on the following domains:
The safety evaluation of nanomaterials, considering the specificity of their physicochemical properties, has been a major concern to prevent unpredictable adverse health effects of those materials. The cytotoxic, genotoxic and epigenetic effects of several classes of nanomaterials has been assessed and mechanistic knowledge has been obtained towards a better understanding of nanomaterials toxicity and early effect biomarkers development. In addition, this research has provided a knowledge basis for decision making in the field of nanosafety. H. Louro and M.J. Silva are INSA representatives at the Portuguese Technical Commission on Nanotechnologies (CT-194, IPQ).
Human biomonitoring/molecular epidemiology
The investigation in this domain is framed by the H2020 European Joint Program Human Biomonitoring Initiative – HBM4EU. Human biomonitoring relies on the measurement of human exposure to chemical substances in the environment by measuring the substances or their metabolites in human specimens. The toxicity and other potential adverse effects of some selected priority substances have been assessed through literature review, data analysis and modeling, in vitro assays and biomarkers analysis in some selected population groups. The results will generate scientific evidence to support decision making in the field of occupational and environmental health,h towards the improvement of national and European health and environment policies.
GENETIC DETERMINANTS OF HUMAN DISEASE
This area includes the investigation of the genetic basis of hereditary conditions (e.g. sexual development pathologies, male infertility, hereditary cancer) to understand the molecular mechanisms of genetic disease. Our current and future research includes molecular studies in two major conditions related with sexual development – idiopathic male infertility, and primary and secondary sex determination/differentiation – both affected by environmental factors. In addition, our research continues with NGS analysis of hereditary breast/ovarian and colorectal cancers. The mutation detection rate is expected to increase in these conditions, and functional assays will contribute to new variants’ pathogenicity evaluation.
Running as PI / Co-PI:
• ToxApp4NanoCELFI – A predictive toxicology approach to characterize potential respiratory effects of functionalized nanocellulose fibers in a co-culture system (PTDC/SAU-PUB/32587/2017) – PI: M.J. Silva; Co-PI: H. Louro
• INGESTNano – Cellular and molecular mechanisms of toxicity of ingested nanomaterials (PTDC/SAU-PUB/29481/2017) – PI: H. Louro; Co-PI: M.J. Silva
• NANOGENOTOX – Safety evaluation of manufactured nanomaterials by characterisation of their potential genotoxic hazard, funded by the Executive Agency for Health and Consumers, 2009 21 01 (2010-2013) – PI of the Portuguese participation: M.J. Silva
• “NANoREG – A common European Approach to the Regulatory Testing of Nanomaterials”, funded under the FP7, theme NMP.21012.1.3-3 NMP.2012.1.3-3 (2013-2016) – PI of the Portuguese participation
Running as Collaborator
• The European Human Biomonitoring Initiative (HBM4EU). Funded under the H2020 (GA 733032); 2017-2021; Coordinator: M. Kolossa-Gehring, UBA, Germany. Coordinator of the Project at INSA: M.J. Silva; Chemical Group Leader of Mycotoxins group. Member of the National Hub for Human Biomonitoring.
• earlyMYCO – early-life exposure to MYCOtoxins and its impact on health (PTDC/MED-TOX/28762/2017) (PI: Paula Alvito, INSA)
• Red Temática CYTED-NANOCELIA – Transferencia Tecnologica sobre Aplicaciones de Nanocelulosa en Iberoamerica; (P316RT0095); 2017 – 2020.
• UPCAST – Unified Platform for CBRN Accident/Attack Scenario management, Projeto em Co-promoção do Programa Operacional PT2020 (SI I&DT) AVISO Nº 08/SI/2015; Coordinator: Tekever, Lisbon; 2016 – 2018.
• NANoREG – A common European Approach to the Regulatory Testing of Nanomaterials, funded under the FP7, theme NMP.21012.1.3-3 NMP.2012.1.3-3; 2013-2017.
• NanoPoliBiofilm – Polymeric therapeutic nanoparticles containing an antimicrobial agent: new opportunities for the development of anti-biofilm agents to treat postetic joint infections (EXCL/CTM-NAN/0166/2012); 2013-2015.
• Mycomix – Exploring the toxic effects of mixtures of mycotoxins in infant food and potential health impact (PTDC/DTP-FTO/0417/2012; 2013-2015.
• HERA – Environmental Risk Assessment of a Contaminated Estuarine Environment: a Case Study (PTDC/SAU-ESA/100107/2008); 2010-2013.
• Environmental Tobacco Smoke Exposure at Portuguese Restaurants, Bars and nightclubs: health effects and early molecular mechanisms underlying respiratory disorders. Fundação Calouste Gulbenkian & MInistry of Health (ACSS); 2008-2013.
• International consortium on “Genetics of Male Infertility”, project – “Genomics of Spermatogenic Impairment”, collaboration with Departments of Genetics, Washington University School of Medicine.
THE MOST RELEVANT SINCE 2013:
Louro H. (2018) Relevance of Physicochemical Characterization of Nanomaterials for Understanding Nano-cellular Interactions. In: Saquib Q., Faisal M., Al-Khedhairy A., Alatar A. (eds) Cellular and Molecular Toxicology of Nanoparticles. Advances in Experimental Medicine and Biology, vol 1048: 124-142. Springer, Cham. ISBN: 978-3-319-72040-1; https://doi.org/10.1007/978-3-319-72041-8_8
Louro H, Bettencourt A, Gonçalves LM, Almeida AJ, Silva MJ. (2015) Role of nanogenotoxicology studies in safety evaluation of nanomaterials. In: Sabu Thomas, Yves Grohens, Neethu Ninan (eds.), Nanotechnology applications in tissue engineering. Elsevier Inc., Oxford, UK, pp.263-288.
Louro H, Pinto M, Vital N, Tavares AM, Costa PM, Silva MJ (2014) The LacZ plasmid based transgenic mouse model: an integrative approach to study the genotoxicity of nanomaterials. In L. María Sierra and Isabel Gaivão (eds.), Genotoxicity and DNA Repair: A Practical Approach, Methods in Pharmacology and Toxicology, Springer-Humana Press, N. Y., USA, pp. 451-478.
Costa PM, Pinto M, Louro H, Gonçalves C, Rodrigo A, Costa MH, Caeiro S, Silva MJ. (2015) An integrative assessment to determine the genotoxic hazard of estuarine sediments: combining cell– and whole–organism responses. In: A. Azqueta, S. Langie, A. Collins (eds.) “30 Years of the Assay: an overview with some insights”. Front. Genet. – Research Topics. ISBN 978-2-88919-649-4 (ebook), pp. 41-52, doi: 10.3389/fgene.2014.00437.
Articles in peer-reviewed journals
Ventura C, Lourenço AF, Sousa-Uva A, Ferreira PJT, Silva MJ (2018). Evaluating the genotoxicity of cellulose nanofibrils in a co-culture of human lung epithelial cells and monocyte-derived macrophages. Toxicology Letters 291:173-183.
Ventura C, Sousa-Uva A, Lavinha J, Silva MJ (2018). Conventional and novel “omics”-based approaches to the study of carbon nanotubes pulmonary toxicity. Environ Mol Mutagen, 59:334-362.
Menezes J, Ventura C, Costa JM, Parreira E, Romão L, Gonçalves J (2017). PROS1 novel splice-site variant decreases protein S expression in patients from two families with thrombotic disease. Clinical Case Reports, 5:2062-2065
Graça D, Louro H, Santos J, Dias K, Gonçalves L, Almeida A, Silva MJ*, Bettencourt A* (2017) Toxicity screening of a novel poly(methylmethacrylate)-Eudragit nanocarrier on L929 fibroblasts. Toxicology Letters, 276: 129-137, doi.org/10.1016/j.toxlet.2017.05.017 (*equal contribution).
Gábelová A, El Yamani N, Alonso T, Buliaková B, Srančíková A, Bábelová A, Runden Pran E, Fjellsbø LM, Elje E, Yazdani M, Silva MJ, Dušinská M (2017) Fibrous shape underlies the mutagenic and carcinogenic potential of nanosilver while surface chemistry affects the biosafety of iron oxide nanoparticles. Mutagenesis, 32: 193-202, doi:10.1093/mutage/gew045.
Caeiro S, Vaz-Fernandes P, Martinho AP, Costa PM, Silva MJ, Lavinha J, Matias-Dias C, Machado A, Castanheira I, Costa MH (2017) Environmental risk assessment in a contaminated estuary: An Integrated weight of evidence approach as a decision support tool. Ocean & Coastal Management, 143: 51-62, doi:10.1016/j.ocecoaman.2016.09.026.
Ferrão J, Silva M, Gonçalves L, Gomes S, Loureiro P, Coelho A, Miranda A, Seuanes F, Batalha Reis A, Pina F, Maia R, Kjöllerström P, Monteiro E, F. Lacerda J, Lavinha J, Gonçalves J, Faustino P (2017). Unusual molecular mechanisms in the origin of alpha-thalassemia. 22nd Congress of the European Hematology Association, 22-25 June, Madrid. E1586. Haematologica 102(s2): 644-645.
Faria R, Silva B, Silva C, Loureiro P, Queiroz A, Fraga S, Esteves J, Mendes D, Fleming R, Vieira L, Gonçalves J, Faustino P (2016). Next-generation sequencing of hereditary hemochromatosis-related genes: Novel likely pathogenic variants found in the Portuguese population. Blood Cells, Molecules and Diseases 61:10–15. http://dx.doi.org/10.1016/ j.bcmd.2016.07.004.
Faria R, Silva B, Silva C, Vieira L, Loureiro P, Gomes S, Gonçalves J, Rivera I, Fraga S, Fleming R, Faustino P (2016). Next-generation sequencing of iron-metabolism related genes in Portuguese patients with iron overload: novel pathogenic genetic variants. European Iron Club, Innsbruck , 7-10 April 2016; Abstract Book P04, Pag 37-38.
Louro H, Pinhão M, Santos J, Tavares A, Vital N, Silva MJ (2016) Evaluation of the cytotoxic and genotoxic effects of benchmark multi-walled carbon nanotubes in relation to their physicochemical properties. Toxicology Letters. 262: 123–13, doi.org/10.1016/j.toxlet.2016.09.016.
Assunção R*, Silva MJ*, Alvito P* (2016) Challenges in risk assessment of multiple mycotoxins in food. World Mycotoxin Jounal, 9: 791-811. DOI 10.3920/WMJ2016.2039 (*equal contribution).
Dekkers S, Oomen AG, Bleeker EAJ, Vandebriel RJ, Micheletti C, Cabellos J, Janer G, Fuentes N,Vazquez-Campos S, Borges T, Silva MJ, Prina-Mello A, Movia D, Nesslany F, Ribeiro AR, Leite PE, Groenewold M, Cassee FR, Sips AJAM, Dijkzeul A, van Teunenbroek T, Wijnhoven SWP (2016) Towards a nanospecific approach for risk assessment. Regulatory Toxicology and Pharmacology 80: 46-59. http://dx.doi.org/10.1016/j.yrtph.2016.05.037.
Collins AR,, Annangi B, Rubio L, Marcos R, Dorn M, Merker C, Estrela-Lopis I, Cimpan MR, Ibrahim M, Cimpan E, Ostermann M, Sauter A, El Yamani N, Shaposhnikov S, Chevillard S, Paget V, Grall R, Delic J, Goñi- de-Cerio F, Suarez-Merino B, Fessard V, Hogeveen KN, Fjellsbø LM, Runden Pran E, Brzicova T, Topinka J, Silva MJ, Leite PE, Ribeiro AR, Granjeiro JM, Grafström R, Prina-Mello A, Dusinska M (2016) High throughput toxicity screening and intracellular detection of nanomaterials. WIREs Nanomed Nanobiotechnol 2016. doi: 10.1002/wnan.1413.
Costa JG, Saraiva N, Guerreiro PS, Louro H, Silva MJ, Miranda JP, Castro M, Batinic-Haberle I, Fernandes AS, Oliveira NG. (2016) Ochratoxin A-induced cytotoxicity, genotoxicity and reactive oxygen species in kidney cells: An integrative approach of complementary endpoints. Food Chem Toxicol., 87:65-76, http://dx.doi.org/10.1016/j.fct.2015.11.018
Pinto M, Louro H, Costa P, Caeiro S, Silva MJ (2015) Exploring the potential interference of estuarine sediment contaminants with the DNA repair capacity of human hepatoma cells. Journal of Toxicology and Environmental Health, Part A: Current Issues. Doi: 10.1080/15287394.2015.1006712.
Bernardo Dias Pereira, Jorge Ralha Portugal, Iris Pereira, João Gonçalves, Luísa Raimundo (2015). X-linked adrenal hipoplasia congenita: clinical and follow-up findings of two kindreds, one with a novel NR0B1 mutation. Arch Endocrinol Metab 59 (2): 181-185. DOI: 10.1590/2359-3997000000032.
A.C. Lima, F. Carvalho, J. Gonçalves, S. Fernandes, P. I. Marques, M. Sousa, A. Barros, S. Seixas, A. Amorim, D. F. Conrad, A. M. Lopes (2015). Rare double sex and mab-3-related transcription factor 1 regulatory variants in severe spermatogenic failure. Andrology. doi: 10.1111/andr.12063.
Tarantini A, Huet S, Jarry G, Lanceleur R, Poul M, Tavares A, Vital N, Louro H, Silva MJ, Fessard V (2014) Genotoxicity of Synthetic amorphous silica nanoparticles in rats following short termexposure. Part 1: Oral route. Environmental and Molecular Mutagenesis. Dec 15. doi: 10.1002/em.21935. [Epub ahead of print].
Costa PM, Pinto M, Louro H, Gonçalves C, Rodrigo A, Costa MH, Caeiro S, Silva MJ (2014) An integrative assessment to determine the genotoxic hazard of estuarine sediments: combining cell– and whole–organism responses. Front. Genet. 2014.5:437. doi:10.3389/fgene.2014.00437.
Louro H, Tavares A, Vital N, Costa PM, Alverca E, Zwart E, de Jong WH, Fessard V, Lavinha J, Silva MJ (2014) Integrated approach to the in vivo genotoxic effects of a titanium dioxide nanomaterials using LacZ plasmid based transgenic mice. Environmental and Molecular Mutagenesis, 55: 500–509.
Dias E, Pinto M, Louro H, Santos T, Antunes S, Pereira P, Silva MJ (2014) Genotoxicity of MicrocystinLR in in vitro and in vivo experimental models. BioMed Res International, 949521(http://dx.doi.org/10.1155/2014/949521).
Pinto M, Rebola M, Louro H, Antunes AMM, José SS, Rocha MR, Silva MJ, Cardoso AS. (2014) Chlorinated polycyclic aromatic hydrocarbons associated with drinking water disinfection: synthesis, occurrence under aqueous chlorination conditions and genotoxic effects. Polycyclic Aromatic Compounds, 34:356371.
Tavares A, Vital N, Costa PM, Alverca E, Zwart E, de Jong WH, Fessard V, Lavinha, Silva MJ. (2014) Integrated approach to the in vivo genotoxic effects of a titanium dioxide nanomaterial using LacZ plasmid-based transgenic mice. Environ Mol Mutag, 55: 500509.
Pinto M, Costa PM, Louro H, Costa MH, Lavinha J, Caeiro S, Silva MJ (2014) Determining oxidative and nonoxidative genotoxic effects driven by estuarine sediment contaminants on a human hepatoma cell line. Science of the Total Environment, 478:2535.
Pinto M, Costa PM, Louro H, Costa MH, Lavinha J, Caeiro S, Silva MJ (2014) Human hepatoma cells exposed to sequential estuarine sediment contaminant extracts permitted the differentiation between cytotoxic and promutagenic fractions. Environ Poll, 185: 141-148.
Catarina M Seabra, Sofia Quental, Ana Paula Neto, Filipa Carvalho, João Gonçalves, João Paulo Oliveira, Susana Fernandes, Mário Sousa, Alberto Barros, António Amorim, Alexandra M Lopes (2014). A novel Alu-mediated microdeletion at 11p13 removes WT1 in a patient with cryptorchidism and azoospermia. Reprod Biomed Online. 29(3):388-91. doi: 10.1016/j.rbmo.2014.04.017. Epub 2014 May 16.
Catarina M Seabra, Sofia Quental, Ana C Lima, Filipa Carvalho, João Gonçalves, Susana Fernandes, Iris Pereira, Júlia Silva, Patrícia I Marques, Mário Sousa, Alberto Barros, Susana Seixas, António Amorim, Alexandra M Lopes (2014). The mutational spectrum of WT1 in male infertility. J Urol. 11. pii: S0022-5347(14)04832-0. doi: 10.1016/j.juro.2014.11.004.
Silva B, Pita L, Gomes S, Gonçalves J, Faustino P (2014). The hepcidin gene promoter c.-1010C>T; −582A>G haplotype modulates serum ferritin in individuals carrying the common H63D mutation in HFE gene. Ann Hematol. 93(12):2063-6 PMID:25015054; DOI: 10.1007/s00277-014-2160-7.
Tavares A, Louro H, Antunes S, Quarré S, Simar S, Temmerman PD, Verleysen E, Mast J, Jensen KA, Norppa H, Nesslany F, Silva MJ (2013) Genotoxicity evaluation of nanosized titanium dioxide, synthetic amorphous silica and multi-walled carbon nanotubes in human lymphocytes, Toxicology in Vitro, 28: 60–69.
Louro H, Borges T, Silva MJ (2013) Nanomateriais manufaturados – Novos desafios para a saúde pública. Revista Portuguesa de Saúde Pública, 31: 145-157.
Tavares, A. M., Alvito, P., Loureiro, S. Louro., H., Silva, M. J. (2013) Multi-mycotoxin determination and in vitro combined cytotoxic effects of aflatoxin M1 and ochratoxin A. World Mycotoxin Journal, 6: 375-388.
Louro A, Peralta L, Soares S, Pereira A, Cunha G, Belchior A, Ferreira L, Monteiro Gil O, Louro H, Pinto P, Rodrigues AS, Silva MJ, Teles P (2013) Human exposure to indoor radon: a survey in the region of Guarda, Portugal. Radiat Prot Dosimetry.154: 237-44.
Pacheco SA, Torres VM, Louro H, Gomes F, Lopes C, Marçal N, Fragoso E, Martins C, Oliveira CL, Hagenfeldt M, Bugalho-Almeida A, Penque D, Simões T (2013) Effects of occupational exposure to tobacco smoke: seeking for a link between environmental exposure and disease. Journal of Toxicology and Environmental Health, Part A, 76:311–327.
Alexandra Lopes; Kenneth I. Aston; Emma E Thompson; Filipa Carvalho; João Gonçalves; Ni Huang; Rune Matthiesen; Michiel J. Noordam; Ines Quintela; Avinash Ramu; Catarina Seabra; Amy B. Wilfert; Juncheng Dai; Jonathan Downie; Susana Fernandes; Xuejiang Guo; Jiahao Shah; Antonio Amorim; Alberto Barros; Angel Carracedo (2013). Human spermatogenic failure purges deleterious mutation load from the autosomes and both sex chromosomes, including the gene DMRT1. PLoS Genet 9(3): e1003349. doi:10.1371/journal.pgen.1003349.
- PToNANO Platform
- Faculdade de Ciências e Tecnologias, Universidade de Coimbra
- Faculdade de Farmácia de Lisboa
- Instituto da Soldadura e Qualidade
- Escola Superior de Tecnologia da Saúde, Instituto Politécnico de LIsboa
- Instituto Português da Qualidade
- Direção Geral da Saúde
- Universidade de Aveiro
- Universidade Aberta
- Faculdade de Ciências Tecnologias, Universidade Nova de Lisboa
- CESAM – Centro de Estudos do Ambiente e do Mar
- MARE – Centro de Ciências do Mar e do Ambiente
- Universidade de Trás-os-Montes e Alto Douro
- Serviço de Medicina da Reprodução – Maternidade Alfredo da Costa
- HBM4EU Consortium
- Maria Dusinska, NILU, Norway
- Hannu Norppa, FIOH, Finland
- NANoREG consortium
- Research groups from several European institutes working in the field of toxicology and environmental health, including (but not limited to) RIVM, The Netherlands; ANSES, France; FIOH, Finland; NILU, Norway.