Publications

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2015
Mendo, Ana Soraia, Sara Figueiredo, Catarina Roma-Rodrigues, Paula A. Videira, Zhen Ma, Mário Diniz, Miguel Larguinho, Pedro Costa, João C. Lima, Armando J. L. Pombeiro, Pedro V. Baptista, and Alexandra R. Fernandes. "Characterization of antiproliferative potential and biological targets of a copper compound containing 4'-phenyl terpyridine." JBIC Journal of Biological Inorganic Chemistry 2 (2015): 935-948. AbstractWebsite

Several copper complexes have been assessed as anti-tumor agents against cancer cells. In this work, a copper compound [Cu(H2O){OS(CH3)2}L](NO3)2 incorporating the ligand 4'-phenyl-terpyridine antiproliferative activity against human colorectal, hepatocellular carcinomas and breast adenocarcinoma cell lines was determined, demonstrating high cytotoxicity. The compound is able to induce apoptosis and a slight delay in cancer cell cycle progression, probably by its interaction with DNA and induction of double-strand pDNA cleavage, which is enhanced by oxidative mechanisms. Moreover, proteomic studies indicate that the compound induces alterations in proteins involved in cytoskeleton maintenance, cell cycle progression and apoptosis, corroborating its antiproliferative potential.

Pedrosa, Pedro, and Pedro Viana Baptista. "Gold and silver nanoparticles for diagnostics of infection." In Nanotechnology in Diagnosis, Treatment and Prophylaxis of Infectious Diseases, edited by Mahendra Rai and Kateryna Kon, 1-18. Elsevier, 2015. Abstract

Nanotechnology in Diagnosis, Treatment and Prophylaxis of Infectious Diseases delivers comprehensive coverage of the application of nanotechnology to pressing problems in infectious disease.
This text equips readers with cutting-edge knowledge of promising developments and future prospects in nanotechnology, paying special attention to microbes that are now resistant to conventional antibiotics, a concerning problem in modern medicine.
Readers will find a thorough discussion of this new approach to infectious disease treatment, including the reasons nanotechnology presents a promising avenue for the diagnosis, treatment, and prophylaxis of infectious diseases.

Vinhas, Raquel, Milton Cordeiro, Fábio Carlos, Soraia Mendo, Alexandra Fernandes, Sara Figueiredo, and Pedro Baptista. "Gold nanoparticle-based theranostics: disease diagnostics and treatment using a single nanomaterial." Journal of Nanobiosensors in Disease Diagnosis (2015): 11-23. AbstractWebsite

Nanotheranostics takes advantage of nanotechnology-based systems in order to diagnose and treat a specific disease. This approach is particularly relevant for personalized medicine, allowing the detection of a disease at an early stage, to direct a suitable therapy toward the target tissue based on the molecular profile of the altered phenotype, subsequently facilitating disease monitoring and following treatment. A tailored strategy also enables to reduce the off-target effects associated with universal treatments and improve the safety profile of a given treatment. The unique optical properties of gold nanoparticles, their ease of surface modification, and high surface-to-volume ratio have made them central players in this area. By combining imaging, targeting, and therapeutic agents in a single vehicle, these nanoconjugates are (ought to be) an important tool in the clinics. In this review, the multifunctionality of gold nanoparticles as theranostics agents will be highlighted, as well as the requirements before the translation of these nanoplatforms into routine clinical practice.

Child, Hannah Winifred, Yulan Hernandez, João Conde, Margaret Mullin, Pedro V. Baptista, Jesus Maria de la Fuente, and Catherine C. Berry. "Gold nanoparticle-siRNA mediated oncogene knockdown at RNA and protein level, with associated gene effects." NANOMEDICINE 10 (2015): 2513-2525. AbstractWebsite

Aims: RNAi is a powerful tool for gene silencing that can be used to reduce undesirable overexpression of oncogenes as a novel form of cancer treatment. However, when using RNAi as a therapeutic tool there is potential for associated gene effects. This study aimed to utilize gold nanoparticles to deliver siRNA into HeLa cells. Results: Knockdown of the c-myc oncogene by RNAi, at the RNA, protein and cell proliferation level was achieved, while also identifying associated gene responses. Discussion: The gold nanoparticles used in this study present an excellent delivery platform for siRNA, but do note associated gene changes. Conclusion: The study highlights the need to more widely assess the cell physiological response to RNAi treatment, rather than focus on the immediate RNA levels.

Franco, Ricardo, Pedro Pedrosa, Fábio Ferreira Carlos, Bruno Veigas, and Pedro Viana Baptista. "Gold Nanoparticles for DNA/RNA-Based Diagnostics." In Handbook of Nanoparticles, 1-25. Berlin: Springer, 2015. Abstract

The remarkable physicochemical properties of gold nanoparticles (AuNPs) have prompted development
in exploring biomolecular interactions with AuNPs-containing systems, pursuing biomedical applications
in diagnostics. Among these applications, AuNPs have been remarkably useful for the development of
DNA/RNA detection and characterization systems for diagnostics, including systems suitable for point of
need. Here, emphasis will be on available molecular detection schemes of relevant pathogens and their
molecular characterization, genomic sequences associated with medical conditions (including cancer),
mutation and polymorphism identification, and the quantification of gene expression.

Pedrosa, Pedro, Raquel Vinhas, Alexandra Fernandes, and Pedro V. Baptista. "Gold Nanotheranostics: Proof-of-Concept or Clinical Tool?" Nanomaterials 5 (2015): 1853-1879. AbstractWebsite

Nanoparticles have been making their way in biomedical applications and personalized medicine, allowing for the coupling of diagnostics and therapeutics into a single nanomaterial—nanotheranostics. Gold nanoparticles, in particular, have unique features that make them excellent nanomaterials for theranostics, enabling the integration of targeting, imaging and therapeutics in a single platform, with proven applicability in the management of heterogeneous diseases, such as cancer. In this review, we focus on gold nanoparticle-based theranostics at the lab bench, through pre-clinical and clinical stages. With few products facing clinical trials, much remains to be done to effectively assess the real benefits of nanotheranostics at the clinical level. Hence, we also discuss the efforts currently being made to translate nanotheranostics into the market, as well as their commercial impact.

Martins, Pedro, João Jesus, Sofia Santos, Luis R. Raposo, Catarina Roma-Rodrigues, Pedro Viana Baptista, and Alexandra Fernandes. "Heterocyclic Anticancer Compounds: Recent Advances and the Paradigm Shift towards the Use of Nanomedicine’s Tool Box." Molecules (2015): 16852-16891. AbstractWebsite

The majority of heterocycle compounds and typically common heterocycle fragments present in most pharmaceuticals currently marketed, alongside with their intrinsic versatility and unique physicochemical properties, have poised them as true cornerstones of medicinal chemistry. Apart from the already marketed drugs, there are many other being investigated for their promising activity against several malignancies. In particular, anticancer research has been capitalizing on the intrinsic versatility and dynamic core scaffold of these compounds. Nevertheless, as for any other promising anticancer drugs, heterocyclic compounds do not come without shortcomings. In this review, we provide for a concise overview of heterocyclic active compounds and families and their main applications in medicine. We shall focus on those suitable for cancer therapy while simultaneously addressing main biochemical modes of action, biological targets, structure-activity relationships as well as intrinsic limitation issues in the use of these compounds. Finally, considering the advent of nanotechnology for effective selective targeting of drugs, we shall discuss fundamental aspects and considerations on nanovectorization of such compounds that may improve pharmacokinetic/pharmacodynamic properties of heterocycles.

Larguinho, Miguel, Sara Figueiredo, Ana Cordeiro, Fábio Ferreira Carlos, Milton Cordeiro, Pedro Pedrosa, and Pedro Viana Baptista. "Nanoparticles for Diagnostics and Imaging." In Frontiers in Nanomedicine, edited by Maria Luisa Bondì Bondì, Chiara Botto and Erika Amore, 3-46. Bentham Science, 2015. Abstractsample.pdf

Nanoparticles possess unique optical and physic-chemical properties that may potentiate applications in biomedicine, in particular in diagnostics, therapy and imaging. Advances on biomolecular diagnostics strategies have greatly focused on single molecule detection and characterization of DNA, RNA or proteins through improved nanoparticle-based platforms. Nanoparticles improve analytical capability when compared to traditional techniques with high resolution and medium-high throughput. Also, particular interest has been directed at SNP detection, gene expression profiles and biomarker characterization through colorimetric, spectrometric or electrochemical strategies.
Molecular imaging has also benefited from the introduction of nanoparticles in standard techniques towards non-invasive imaging procedures that can be used to highlight regions of interest, allowing the characterization of biological processes at the cellular and/or molecular level. Several imaging modalities are associated with low sensitivity, an issue that can be tackled by the use of probes, e.g. contrast agents for X-ray and magnetic resonance imaging, radiolabelled molecules for nuclear medicine. Furthermore, nanoparticles can be used as vehicles that deliver specifically these contrast agents, leading to overcome the limitations of conventional modalities.
This chapter will discuss the use of nanoparticles in biomolecular recognition and imaging applications, focusing those already being translated into clinical settings. Current knowledge will be addressed as well as its evolution towards the future of nanoparticle-based biomedical applications.

Martins, Pedro, Mara Marques, Lidia Coito, Armando Pombeiro, Pedro V. Baptista, and Alexandra R. Fernandes. "Organometallic Compounds in Cancer Therapy: Past Lessons and Future Directions." ANTI-CANCER AGENTS IN MEDICINAL CHEMISTRY 9 (2015). AbstractWebsite

Over the past few years, modern medicinal chemistry has evolved towards providing us new and alternative chemotherapeutic compounds with high cytotoxicity towards tumor cells, alongside with reduced side effects in cancer patients. Organometallic compounds and their unique physic-chemical properties typically used in homogenous catalysis are now being translated as potential candidates for medical purposes. Their structural diversity, ligand exchange, redox and catalytic properties make them promising drug candidates for cancer therapy. Over the last decade this area has witnessed a steady growth and a few organometallic compounds have in fact already entered clinical trials, emphasizing its increasing importance and clinical relevance. Here we intend to stress out the different applications of organometallic compounds in medicine with emphasis on cancer therapy, as well as address setbacks regarding formulation issues, systemic toxicity and off-target effects. Advantages over classical coordination metal complexes, their nanovectorisation and specific molecular targets are also discussed.

Restani, Rita B., Joao Conde, Rita F. Pires, Pedro Martins, Alexandra R. Fernandes, Pedro V. Baptista, and Vasco D. B. Bonifacio. "POxylated Polyurea Dendrimers: Smart Core-Shell Vectors with IC50 Lowering Capacity." MACROMOLECULAR BIOSCIENCE 15 (2015): 1045-1051. AbstractWebsite

The design and preparation of highly efficient drug delivery platforms using green methodologies is at the forefront of nanotherapeutics research. POxylated polyurea dendrimers are efficiently synthesized using a supercritical-assisted polymerization in carbon dioxide. These fluorescent, pH-responsive and water-soluble core-shell smart nanocarriers show low toxicity in terms of cell viability and absence of glutathione depletion, two of the major side effect limitations of current vectors. The materials are also found to act as good transfection agents, through a mechanism involving an endosomal pathway, being able to reduce 100-fold the IC50 of paclitaxel.

McCully, Mark, Yulan Hernandez, João Conde, Pedro V. Baptista, Jesus M. de la Fuente, Andrew Hursthouse, and David St. "The significance of the balance between intracellular glutathione and polyethylene glycol (PEG) for successful siRNA release from gold nanoparticles." Nano Research (2015). AbstractWebsite

The therapeutic promise of small interfering RNAs (siRNAs) for specific gene silencing is dependent on the
successful delivery of functional siRNAs to the cell cytoplasm. Their conjugation to an established delivery
platform, such as gold nanoparticles, offers a huge potential for treating diseases and advancing our
understanding of cellular processes. The success or failure is dependent on both the uptake of the nanoparticlesinto the cells and subsequent intracellular release of the functional siRNA. In this paper, utilising gold nanoparticle siRNA-mediated delivery against C-MYC, we aimed to determine if we could achieve knockdown in a cancer cell line with low levels of intracellular glutathione, and determine the influence, if any, of polyethylene glycol (PEG) ligand density on knockdown, with a view to determine the optimal nanoparticle
design to achieve C-MYC knockdown. We demonstrate that, regardless of the PEG density, knockdown in cells with relatively low glutathione levels can be achieved, and also the possible effect of steric hindrance in terms of PEG on the availability of the siRNA for cleavage in the intracellular environment. Gold nanoparticle uptake was demonstrated via transmission electron microscopy and mass spectroscopy, whilst knockdown was determined at the protein and physiological levels (cells in S-phase) by in-cell westerns and BrdU incorporation, respectively.

2016
Vinhas, Raquel, Claudia Correia, Patricia Ribeiro, Alexandra Lourenco, Aida Botelho de Sousa, Alexandra R. Fernandes, and Pedro V. Baptista. "Colorimetric assessment of BCR-ABL1 transcripts in clinical samples via gold nanoprobes." Analytical and Bioanalytical Chemistry 408 (2016): 5277-5284. Abstract

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Cordeiro, Milton, Fabio Ferreira Carlos, Pedro Pedrosa, Antonio Lopez, and Pedro Viana Baptista. "Gold Nanoparticles for Diagnostics: Advances towards Points of Care." Diagnostics 6 (2016). Abstract

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Lenis-Rojas, O. A., A. R. Fernandes, C. Roma-Rodrigues, P. V. Baptista, F. Marques, D. Perez-Fernandez, J. Guerra-Varela, L. Sanchez, D. Vazquez-Garcia, M. Lopez Torres, A. Fernandez, and J. J. Fernandez. "Heteroleptic mononuclear compounds of ruthenium(II): synthesis, structural analyses, in vivo antitumor activity and in vivo toxicity on zebrafish embryost." Dalton Transactions 45 (2016): 19127-19140. Abstract

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Martins, Marta, Pedro V. Baptista, Ana Soraia Mendo, Claudia Correia, Paula Videira, Antonio S. Rodrigues, J. Muthukumaran, Teresa Santos-Silva, Ana Silva, Fatima M. C. Guedes da Silva, Joana Gigante, Antonio Duarte, Malgorzata Gajewska, and Alexandra R. Fernandes. "In vitro and in vivo biological characterization of the anti-proliferative potential of a cyclic trinuclear organotin(IV) complex." Molecular Biosystems 12 (2016): 1015-1023. Abstract

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Roma-Rodrigues, Catarina, Cynthia Alves-Barroco, Luis R. Raposo, Mafalda N. Costa, Elvira Fortunato, Pedro Viana Baptista, Alexandra R. Fernandes, and Ilda Santos-Sanches. "Infection of human keratinocytes by Streptococcus dysgalactiae subspecies dysgalactiae isolated from milk of the bovine udder." Microbes and Infection 18 (2016): 290-293. Abstract

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Luisa Corvo, M., Ana Soraia Mendo, Sara Figueiredo, Rogerio Gaspar, Miguel Larguinho, Fatima M. C. Guedes da Silva, Pedro Viana Baptista, and Alexandra R. Fernandes. "Liposomes as Delivery System of a Sn(IV) Complex for Cancer Therapy." Pharmaceutical Research 33 (2016): 1351-1358. Abstract

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Mendes, Rita, Barbara Carreira, Pedro V. Baptista, and Alexandra R. Fernandes. "Non-small cell lung cancer biomarkers and targeted therapy - two faces of the same coin fostered by nanotechnology." Expert Review of Precision Medicine and Drug Development 1 (2016): 155-168. Abstract

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Vinhas, Raquel, Anna Tolmatcheva, Rafaela Canto, Patricia Ribeiro, Alexandra Lourenco, Aida Botelho de Sousa, Pedro V. Baptista, and Alexandra R. Fernandes. "A novel mutation in CEBPA gene in a patient with acute myeloid leukemia." Leukemia & Lymphoma 57 (2016): 711-713. Abstract

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Roma-Rodrigues, Catarina, Amelie Heuer-Jungemann, Alexandra R. Fernandes, Antonios G. Kanaras, and Pedro V. Baptista. "Peptide-coated gold nanoparticles for modulation of angiogenesis in vivo." International Journal of Nanomedicine 11 (2016): 2633-2639. Abstract

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Baptista, Pedro Viana. "Precision nanomedicine in cancer: how far are we from personalization?" Expert Review of Precision Medicine and Drug Development 1 (2016): 227-228. Abstract

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Cordeiro, Milton, Leticia Giestes, Joao Carlos Lima, and Pedro Baptista. "BioCode gold-nanobeacon for the detection of fusion transcripts causing chronic myeloid leukemia." Journal of Nanobiotechnology 38 (2016). AbstractWebsite

Background
Gold-nanobeacons (Au-nanobeacons) have proven to be versatile systems for molecular diagnostics and therapeutic actuators. Here, we present the development and characterization of two gold nanobeacons combined with Förster resonance energy transfer (FRET) based spectral codification for dual mode sequence discrimination. This is the combination of two powerful technologies onto a single nanosystem.

Results
We proved this concept to detect the most common fusion sequences associated with the development of chronic myeloid leukemia, e13a2 and e14a2. The detection is based on spectral shift of the donor signal to the acceptor, which allows for corroboration of the hybridization event. The Au-nanobeacon acts as scaffold for detection of the target in a homogenous format whose output capability (i.e. additional layer of information) is potentiated via the spectral codification strategy.

Conclusions
The spectral coded Au-nanobeacons permit the detection of each of the pathogenic fusion sequences, with high specificity towards partial complementary sequences. The proposed BioCode Au-nanobeacon concept provides for a nanoplatform for molecular recognition suitable for cancer diagnostics.

Vinhas, Raquel, Milton Cordeiro, Pedro Pedrosa, Alexandra R. Fernandes, and Pedro V. Baptista. "Current trends in molecular diagnostics of chronic myeloid leukemia." Leukemia & Lymphoma (2016): 1-14. AbstractWebsite

Nearly 1.5 million people worldwide suffer from chronic myeloid leukemia (CML), characterized by the genetic translocation t(9;22)(q34;q11.2), involving the fusion of the Abelson oncogene (ABL1) with the breakpoint cluster region (BCR) gene. Early onset diagnosis coupled to current therapeutics allow for a treatment success rate of 90, which has focused research on the development of novel diagnostics approaches. In this review, we present a critical perspective on current strategies for CML diagnostics, comparing to gold standard methodologies and with an eye on the future trends on nanotheranostics.

Conde, Joao, Furong Tian, Jesus M. de la Fuente, and Pedro V. Baptista. "Editorial: Cancer Nanotheranostics: What Have We Learned So Far?" Frontiers in Chemistry 71 (2016).Website
Cordeiro, Mílton, Fábio Ferreira Carlos, Pedro Pedrosa, António Lopez, and Pedro Viana Baptista. "Gold Nanoparticles for Diagnostics: Advances towards Points of Care." Diagnostics 6 (2016): 43. AbstractWebsite

The remarkable physicochemical properties of gold nanoparticles (AuNPs) have prompted developments in the exploration of biomolecular interactions with AuNP-containing systems, in particular for biomedical applications in diagnostics. These systems show great promise in improving sensitivity, ease of operation and portability. Despite this endeavor, most platforms have yet to reach maturity and make their way into clinics or points of care (POC). Here, we present an overview of emerging and available molecular diagnostics using AuNPs for biomedical sensing that are currently being translated to the clinical setting.