Bioneedles for tailored pain-free drug delivery

 

AREA OF KNOWLEDGE

Life Sciences Panel

 

GROUP OF DISCIPLINE

Biotechnology, Bioinformatics, Pharmaceuticals, Food Technology

 

GROUP LEADER

Prof. Ana Cecília Afonso Roque
cecilia.roque@fct.unl.pt

 

RESEARCH PRODUCT / RESEARCH GROUP

The Biomolecular Engineering Lab @UCIBIO @i4HB (NOVA School of Science and Technology), was established in 2008 by Prof. Cecília Roque and is composed by a multidisciplinary team with expertise in Applied and Computational Chemistry, Biotechnology, Biomedical Engineering, Chemical Engineering and Materials Science. Our group is dedicated to MINIMAL BIOMIMETIC SYSTEMS, namely combining designed molecular recognition agents with advanced functional materials, which together find applications in Biotechnology and Biomedicine.

https://biomolecular-engineering-lab.pt/

 

POSITION DESCRIPTION

 

Research Project / Research Group Description

This is a collaborative research project between two labs @UCIBIO, FCT-NOVA (www.ucibio.pt), combining the unique know-how on stimuli-responsive protein-based materials (Roque´s lab) with the excellent mechanical properties of bio-based and biodegradable polymers produced by sustainable approaches (Reis´ lab).

Microneedles are excellent alternatives for delivery as they are simple to operate, painless and with high patient compliance. Still, there are several challenges associated with the wide implementation of microneedle devices, due to the need to develop biocompatible materials with tuneable release profiles. The goal is to design, develop and test microneedles for the tailor-made transdermal delivery of therapeutic agents.

In this work we will use biopolymers that are obtained from microbial transformation of wastes into high value products. Reis lab has unique experience on the biological valorization of wastes and by-products from agro-food and industrial processes for the production of microbial biopolymers by single and mixed culture systems. One of the key developments of the lab, that will be explored in this project, is the ability to tune the biotechnological production process to obtain biopolymers with distinct and tunable mechanical properties, a key aspect to obtain flexible, yet robust microneedle devices. This can be obtained by combining feedstocks with different compositions and/or imposing key reactor operational conditions and microbial cultures. To endow the microneedle devices with stimuli-responsive properties, for the on-demand release of drugs according to physiological cues, the know-how from Roque´s lab on stimuli-responsive and autonomously actuated protein-based materials will be critical. This will confer a simple, sustainable and biocompatible synergetic solution for the development of novel microneedle devices.

 

Job position description:

This is a position for a PhD student to develop his/her research project at Roque and Reis labs. The PhD student will be responsible for the planning and daily execution of the research project, participate in lab management activities, and in training, dissemination and communication activities. The student should have ideally a background in biotechnology/biological engineering fields.

 

OTHER RELEVANT WEBSITES

This project is in collaboration with the Biochemical Engineering (BIOENG) group, led by Prof. Reis, that develops research (both fundamental and applied) in the areas of Industrial and Environmental Biotechnology, with main focus on on the valorization of wastes/by-products through the production and characterization of microbial biopolymers such as Polyhydroxyalkanoates (PHAs),Polysaccharides and Bulk Chemicals. BIOENG is an interdisciplinary group with strong collaborations with Industries and research centers at national and international level.

https://sites.fct.unl.pt/bioeng/home

 

 

Carbon monoxide modulation of Brain Homeostasis: targeting Peroxisome Metabolism

 

AREA OF KNOWLEDGE

Life Sciences Panel

 

GROUP OF DISCIPLINE

Human Biology, Microbiology, Molecular Biology, Genetics, Cell Biology, Genomics and Proteomics, Biochemistry, Basic Neuroscience

 

GROUP LEADER

Prof. Helena Luisa de Araujo Vieira
hl.vieira@fct.unl.pt

 

RESEARCH PRODUCT / RESEARCH GROUP

Description of the Research group. For a more complete presentation of the results generated by the group and concerning the present research proposal, please see the link Group History Vieira

https://www.ucibio.pt/research-groups/lab/molecular-mechanisms-disease

 

POSITION DESCRIPTION

 

Research Project / Research Group Description

Carbon monoxide (CO) is endogenously produced by the stress response heme oxygenase enzyme and presents homeostatic roles: anti-inflammatory and cytoprotective in several tissues including the brain (1). In the last decade, Vieira’s group has been exploring CO as a cytoprotective molecule in the brain, showing CO’s anti-apoptotic and anti-inflammatory role in neurons, astrocytes and microglia. Moreover, we have been pioneer in demonstrating that CO modulates cell metabolism, namely increasing oxidative phosphorylation and mitochondrial metabolism (Group History Link).

Herein the main objective is to explore peroxisome function in CO-induced cytoprotection, in particular how peroxisome-mediated lipid metabolism modulates mitochondrial metabolism and inflammatory response. Despite the physiopathological role of CO in mitochondria has been extensively studied, nothing is known about peroxisome, which is a far less explored organelle but with a key impact on lipid metabolism.

Lipid metabolism is important for microglial control of inflammation and phagocytosis control (2). In addition, peroxisomes are critical for lipid oxidation and their biogenesis is controlled by PGC-1alpha, which also activates mitochondrial biogenesis and is induced by CO (3). Thus, it can be hypothesized that CO stimulates very long and long-chain fatty acid oxidation into short fatty acids that fuel mitochondrial metabolism and control inflammation.

CO modulation of peroxisome metabolism will be studied in key cell types with impact on brain homeostasis: microglia and blood brain barrier (BBB) endothelial cells. In fact, several neurological diseases (namely vascular dementia, stroke or neurodegenerative diseases) present common features, including BBB disruption and chronic or exacerbated neuroinflammation, with microglial dysfunction (4,5).

1- DOI 10.1111/bph.12729

2- DOI 10.3390/ijms20163877

3- DOI 10.1016/j.redox.2020.101470

4- DOI 10.1038/nri.2017.125

5- DOI 10.1152/physrev.00050.2017

 

Job position description

BBB is constituted by cerebral microvascular endothelial cell, astrocytes and pericytes and present unique characteristics with higher selectivity and limited permeability for the maintenance of brain homeostasis. BBB protects the brain against neurotoxic plasma factors, blood cells and pathogens. Microglia control phagocytosis and the production of pro- and anti-inflammatory cytokines for damage resolution and maintenance of homeostasis. Nevertheless, under exacerbated or chronic neuroinflammation, microglia can become neurotoxic.

The focus of this research will be the unexplored role of CO on lipid metabolism and peroxisome activity and its impact on BBB and microglial functions.

 

First aim will be the study of peroxisomes function and biogenesis in microglia, astrocytes and brain microvasculature endothelial cells (BBB). Peroxisome population and activity will be quantified by the levels of specific peroxisomal proteins (ACOX1, PEX and PGC-1alpha). Fatty acid oxidation will be analyzed by NMR in collaboration with Dr Jones. Preliminary NMR data indicate an increase of the short fatty acid acetate levels during inflammatory response of microglia. Lipid metabolism will be studied along with other metabolic pathways, such as oxidative phosphorylation, in order to disclose the cross-talk between mitochondria-peroxisome.

Secondly, the functional impact of CO modulation of lipid metabolism will be explored in microglial function and BBB permeabilization.

It is well established that CO is anti-neuroinflammatory and we have preliminary data pointing to CO regulation of microglial phagocytosis of dead neurons (clearance of toxic factors). Likewise, it was shown that CO protects BBB permeabilization. Thus, cell culture and in vivo models will be used for exploring whether lipid metabolism and peroxisomes play a role in these biological protective effects of CO.

Please see in the link – Techniques and Collaborations for the techniques, models and the network of collaborations.

 

OTHER RELEVANT WEBSITES

Superviser Scopus link: https://www.scopus.com/authid/detail.uri?authorId=57197339848

Research group History about Carbon monoxide and Neurobiology link: https://www.dropbox.com/s/mvcanji4deaajp1/GroupHistoryVieira.pdf?dl=0

Techniques description and network of collaborators involved in the project: https://www.dropbox.com/s/evhlz9994rrrwvj/Techniques_Collaborations.pdf?dl=0

 

 

Structural and functional studies on G-quadruplex/nucleolin complex and its role in lung cancer

 

AREA OF KNOWLEDGE

Life Sciences Panel

 

GROUP OF DISCIPLINE

Human Biology, Microbiology, Molecular Biology, Genetics, Cell Biology, Genomics and Proteomics, Biochemistry, Basic Neuroscience

 

GROUP LEADER

Prof. Eurico Cabrita
ejc@fct.unl.pt

 

RESEARCH PRODUCT / RESEARCH GROUP

The Biomolecular Structure and interactions by NMR research Lab interests are related to the study of intermolecular interactions in biological and chemical systems by solution state NMR spectroscopy. The main contributions are in the understanding of molecular recognition processes in different contexts, namely, protein-ligand interactions in drug discovery.

https://www.ucibio.pt/research-groups/lab/biomolecular-structure-and-interactions-nmr

 

POSITION DESCRIPTION

 

Research Project / Research Group Description

This collaborative research project between UCIBIO and CICS-UBI combines unique know-how on structural molecular biology to study proteins and protein-ligand complexes, (Cabrita and Romao labs) with biophysical and cellular assays to check G-quadruplex (G4) aptamers formation, its recognition by a specific target and the implications in lung cancer (LC) (Cruz lab).

Nucleolin (NCL) is highly expressed by surface cancer cells and recent studies suggest an inhibitory effect in tumour growth and gene expression after G4-NCL complex formation [1]. Relevant examples are NCL complex formation with G4 of NHE III1 region of the c-MYC promoter repressing MYC transcription, mRNA stabilization and protection from degradation. NCL has distinct functions depending on different cellular compartments in cancer cells, but the structure associated with its subcellular expression levels requires further clarification.

DNA/RNA G4 aptamers represent a novel therapeutic strategy in cancer. They combine binding efficiency and selectivity for unrelated targets, like NCL, and can exhibit high conformational stability after ligand binding.

Based on previous studies the goal of this project is to study the role of G4-NCL complex formation and its implications in NCL expression in LC cell models.

We will use known cancer-specific DNA/RNA aptamers with the potential to form stable G4 for NCL binding [2].

One of the key developments proposed is to investigate the preclinical impact of how the formation of G4–NCL complex is related with NCL expression and LC, by determining the complex 3D structure, and evaluating tumour suppressor genes, transcription, protein repression and growth factors whose angiogenic/tumorigenic activities are mediated by NCL. To accomplish this, we will use cellular lung cancer models and peripheral blood mononuclear cells (PBMCs) co-culture from patients with LC.

[1] DOI:10.1016/j.tcb.2022.03.003

[2] DOI:10.1016/j.jpba.2022.114757

 

Job position description

This is a position for a PhD student to develop his/her research project at Romao & Cabrita labs at FCT-NOVA/UCIBIO (www.fct.unl.pt/en www.ucibio.pt) with some stays at the Cruz lab (https://www.ubi.pt/sites/cics/en). The PhD student will be responsible for the planning and daily execution of the research project, participate in lab management activities, and in training, dissemination and communication activities. The student should ideally have a background in biochemistry/biophysics or related areas.

 

Supervisor:

Eurico Cabrita: orcid.org/0000-0002-0720-2751

 

Co-supervisors:

Maria Joao Romao:  orcid.org/0000-0002-3004-0543

Carla Cruz: orcid.org/0000-0001-6630-1242

 

OTHER RELEVANT WEBSITES

The macromolecular crystallography research Lab at UCIBIO-FCT-NOVA, headed by Maria Joao Romao, uses structural molecular biology tools to study proteins and protein-ligand complexes, using molecular biology techniques, X-ray Crystallography, Cryo-EM, Small Angle X-ray Scattering (SAXS) and other biophysical methods.

https://www.ucibio.pt/research-groups/lab/xtal-macromolecular-crystallography

G-quadruplex research group at CICS-UBI is focused on the optimization of biophysical/cellular and molecular assays to study DNA/RNA G-quadruplex structures, aptamers and protein targets towards cancer.

https://gquadruplex.wixsite.com/ccruz

 

 

Tackling drivers of malignant transformation of polyps to colorectal cancer via Molecular Profilling and Nanotheranostics

 

AREA OF KNOWLEDGE

Life Sciences Panel

 

GROUP OF DISCIPLINE

Biotechnology, Bioinformatics, Pharmaceuticals, Food Technology

 

GROUP LEADER

Prof. Alexandra Núncio de Carvalho Ramos Fernandes
ma.fernandes@fct.unl.pt

 

RESEARCH PRODUCT / RESEARCH GROUP

The major goals of Human Genetics and cancer therapeutics group (co-supervisor) are the identification and validation of cancer diagnostics biomarkers and the development of novel combinatorial cancer therapies, using in vitro 2D (cell monocultures) and 3D (spheroids) cellular models that simulate tumor microenvironment, patient´s derived cells or peripheral blood, or even in vivo models.

https://ucibio.pt/research-groups/lab/human-genetics-and-cancer-therapeutics

 

POSITION DESCRIPTION

 

Research Project / Research Group Description

The identification of genes and pathways that drive formation of invasive colorectal cancers (CRC) has been the central focus of several large-scale genomics efforts. However, most studies have focused on bulk profiling of advanced stage CRC and have largely ignored premalignant lesions. As a result, understanding the progression of phenotypic changes that occur during the transition from normal to precancerous polyps which can subsequently give rise to CRCs, together with the molecular drivers of this transformation, is of utmost importance for early diagnostics and improved therapeutics regimens.

Gold nanoparticles (AuNPs) have been extensively explored in combined diagnostic and therapeutic concepts – theranostics - due to their unique properties that are well suited for the detection and delivery of cargo (siRNAs, drugs, antibodies targeting specific biomarkers) to the desired cells.

Polyps and surrounding normal tissue from patients with familial adenomatous polyposis (with informed written consent) who undergo surgical colectomies, will be used to isolate single-cell subsets (immune, stromal (particularly, fibroblast subpopulations) and epithelial cell types) that will be further used to develop primary cell cultures (part A). RNA-seq will be used to understand the molecular drivers of malignant transition (part B). These molecular drivers will be further used to tailor personalized nanosystems for improved theranostics applications (part C).

This project will be carried out @Human Genetics & Cancer Therapeutics Group led by Alexandra R Fernandes (ARF) (supervisor) and @Nanomedicine Group led by Pedro V Baptista (PVB) (co-supervisor), both part of UCIBIO FCT-NOVA (www.ucibio.pt). Recently UCIBIO has integrated the successful application to Associated Laboratory in Bioeconomy and Health, whose strategic line focused on development of innovative (nano)therapeutics would profit from the current PhD project.

 

Job position description

The project aims at the understanding of the molecular players, by using RNA-sequencing, that drives malignant transition from premalignant lesions to CRC. These molecular players are important for developing precision diagnostics and therapeutics that can assist clinicians in an early detection and management of patients.  Multifunctional gold nanoparticles will be used to tailor this strategy. The host groups have extensive expertise in the development of novel Diagnostic and Therapeutic strategies to fight cancer (https://orcid.org/0000-0003-2054-4438; https://orcid.org/0000-0001-5255-7095).

This project builds upon expertise of the host groups – Molecular cell biology and models & nanoconjugates – where the suitable candidate shall be integrated. The candidate should have a strong background in biomolecular aspects of the cell (biology, biochemistry, biomedicine, biotechnology). The proposed work relates to different areas of research – from molecular and cell biology to nanomedicine.

 

OTHER RELEVANT WEBSITES

This project is in collaboration with the group of Nanomedicine@FCT - created and developed by Prof. Pedro V Baptista in 2003 - in the field of biofunctionalization of nanoparticles with targeting and silencing moieties and their application in drug delivery and/or gene silencing, in the assessment of toxicology of nanoparticles in in vivo models, in the development of nanobiosensors for DNA/RNA biomolecular recognition studies. At the intersection of Molecular Genetics and Nanotechnology, we have focused our research on the use of noble metal nanoparticles for new diagnostics and therapeutics platforms

https://ucibio.pt/research-groups/lab/nanomedicine

 

 

Targeting a ‘toolbox’ of human microbiome O-Glycan-Selective enzymes towards new approaches in cancer research

 

AREA OF KNOWLEDGE

Life Sciences Panel

 

GROUP OF DISCIPLINE

Human Biology, Microbiology, Molecular Biology, Genetics, Cell Biology, Genomics and Proteomics, Biochemistry, Basic Neuroscience

 

GROUP LEADER

Prof. Maria Angelina de Sá Palma
angelina.palma@fct.unl.pt

 

RESEARCH PRODUCT / RESEARCH GROUP

The Functional Glycobiology Lab focuses on understanding carbohydrate (glycan)-protein interactions for discovering novel biomarkers and developing glycan-based therapies and biotechnological applications. Through an integrative approach, the power of high-throughput glycan microarray technologies is combined with structural biology to identify functional glycans for proteins at a glycomics scale and uncover the molecular determinants of glycan recognition by endogenous lectins and in host-human microbiome interactions.

https://www.ucibio.pt/research-groups/lab/glycolab-functional-glycobiology

 

POSITION DESCRIPTION

 

Research Project / Research Group Description

The fellow will develop research under the FCT-funded GlycOSELECT project that aims to discover novel enzymes with O-glycan selectivity (O-glycopeptidases) from the human microbiome as an analytical ‘toolbox’ for molecular and functional analysis of cancer-associated glycoproteins. The project gathers an interdisciplinary team with complementary expertise in molecular biology, protein biochemistry, structural biology, chemoenzymatic synthesis, technologies in glycomics, glycoproteomics and cancer cell biology.

Mucin-type O-glycosylation is a post-translational modification of cell-surface and secreted proteins, which is associated with tissue- and cell-specific activities, including cell-attachment of commensal and pathogenic microbes and development and progression of cancer. Alterations of O-glycosylation are known to modulate more aggressive cancer phenotypes and tumour associated O-glycans are molecules with high potential for development of clinical biomarkers. Research on human microbiome uncovered O-glycopeptidases of commensal and pathogenic bacteria that have adapted to the human gastrointestinal tract, a mucin O-glycoprotein-rich ecosystem.

The project hypothesis is that these O-glycopeptidades exhibit different O-glycan and peptide-backbone selectivity to target mucin-type O-glycoproteins, which will translate into functionally diverse enzymes to build a toolbox targeting normal vs tumour associated O-glycans. The proposed objectives are: i) to characterise O-glycan specificity and substrate selectivity of novel microbiome-associated O-glycopeptidases; ii) to apply the O-glycopeptidases in analysis of cancer-associated glycoproteins in cancer cell models and patient tissues.

 

Job position description

The work will be mainly carried out at the Functional Glycobiology Lab (Supervisor, A Palma) and Macromolecular Crystallography Lab (Co-Supervisor, AL Carvalho,) at UCIBIO-NOVA. There will be programmed stays at the collaborator’s Institutions: i3S-Porto (PT), Imperial College London (UK) and Umea University (SWE). Therefore, mobility will be supported. The Fellow shall be integrated in the scientific team and will participate in all activities of the GlycOSELECT project and will be responsible to develop an interdisciplinary research plan in Molecular Glycosciences. The plan will be mainly focused in applying glycome-based microarray approaches and structural biology for the rational design of the O-glycopeptidases, and in the setting-up of functional assays for their application in analysis of cancer-associated glycoproteins. The student should have a background in biochemistry/chemistry/biology or related areas.

 

OTHER RELEVANT WEBSITES

The Macromolecular Crystallography Lab uses structural molecular biology tools to study proteins and protein-ligand complexes, using X-ray Crystallography, Cryo-Electron Microscopy, Small Angle X-ray Scattering (SAXS) and other biophysical methods. Main research areas include: mechanistic studies and identification of ligands, at atomic level; Drug metabolism and ligand discovery; Glycan-protein and protein-protein interactions; Novel approaches for improved protein crystallization.

https://www.ucibio.pt/research-groups/lab/xtal-macromolecular-crystallography

 

 

The Impact of human Aldehyde Oxidase on Drug Metabolism – Structural and Functional studies

 

AREA OF KNOWLEDGE

Life Sciences Panel

 

GROUP OF DISCIPLINE

Human Biology, Microbiology, Molecular Biology, Genetics, Cell Biology, Genomics and Proteomics, Biochemistry, Basic Neuroscience

 

GROUP LEADER

Prof. Maria Joao Romao
mjr@fct.unl.pt

 

RESEARCH PRODUCT / RESEARCH GROUP

The macromolecular crystallography research Lab at UCIBIO-FCT-NOVA, headed by Maria Joao Romao, uses structural molecular biology tools to study proteins and protein-ligand complexes, using molecular biology techniques, X-ray Crystallography, Cryo-EM, Small Angle X-ray Scattering (SAXS) and other biophysical methods.

https://www.ucibio.pt/research-groups/lab/xtal-macromolecular-crystallography

 

POSITION DESCRIPTION

 

Research Project / Research Group Description

Human aldehyde oxidase (hAOX1) is a complex cytosolic metalloenzyme, with no clear physiological function yet assigned, but known to play an essential role in phase-I drug and xenobiotic metabolism. The lack of understanding of the mechanisms underlying the diverse panoply of reactions catalyzed by hAOX1 and its multiple substrates has led to the failure of clinical trials, raising Academia and Industry concern [1]. To worsen the problem, the differences between human and rodent enzymes leads to difficulties when trying to comprehend AOX drug metabolism in standard preclinical animal models. AOXs have a complex evolutionary profile being species-specific and, while humans possess only one active AOX gene, rodents express 4 isoforms with differing tissue and substrate specificities. These disparities lead to challenges and difficulties when trying to comprehend hAOX1 drug metabolism in preclinical animal models, and this has often resulted in unpredicted hepatic clearance and ultimately led to clinical trials failures.

The first crystal structures of hAOX1 as well as of mouse AOX3 (Romao lab) enabled the discovery of molecular determinants for substrate binding and reactivity, as well as unexpected inhibition modes with implications, namely in drug-drug interactions. However, many aspects remain to be uncovered, and the emergence of novel catalytic reactions means that many more drugs in the market and drug candidates are expected to be hAOX1 substrates, and deeper knowledge is required.  A step forward was possible in an integrated study employing crystallography, NMR and kinetics that led to a first experimental method for predicting putative substrates and inhibitors [2].

Further mechanistic and structural studies are however still required to understand the complexities of this enzyme, to unravel its true physiological role and to accelerate drug discovery campaigns.

1- 10.1016/j.ccr.2018.04.006; 10.1074/jbc.REV119.007741

2- 10.1021/acs.jmedchem.1c01125

 

Job position description

This is a position for a PhD student to develop his/her research project at the Romao & Cabrita labs at FCT-NOVA University Lisbon www.fct.unl.pt/en  / Research Unit UCIBIO (www.ucibio.pt)

The work will involve an integrated structural biology approach involving complementary biochemical, biophysical, and structural studies (NMR, X-ray crystallography, cryo-EM, TR-SSX) that will be used to unravel the mechanisms of action of hAOX1 (free and in complex with substrates or inhibitors) contributing to enhancing the knowledge of AOX metabolism in humans with impact on drug discovery.

Short stays abroad are expected, namely at synchrotron labs in Europe (ESRF, ALBA or DESY) for X-ray data collection experiments as well as for Time Resolved Serial Synchrotron Crystallography (TR-SSX at DESY) (10.1107/s2059798321011621) through an on-going collaboration with Arwen Pearson (https://www.physik.uni-hamburg.de/en/inf/ag-pearson.html)

The PhD student will be responsible for the planning and daily execution of the research project, participate in lab management activities, and in training, dissemination, and communication activities. The student should ideally have a background in biochemistry/biophysics or related areas.

 

Supervisor:

Maria Joao Romão:  orcid.org/0000-0002-3004-0543

 

Co-supervisor:

Eurico Cabrita: orcid.org/0000-0002-0720-2751

 

OTHER RELEVANT WEBSITES

The Biomolecular Structure and interactions by NMR research Lab interests are related to the study of intermolecular interactions in biological and chemical systems by solution state NMR spectroscopy. The main contributions are in the understanding of molecular recognition processes in different contexts, namely, protein-ligand interactions in drug discovery.

https://www.ucibio.pt/research-groups/lab/biomolecular-structure-and-interactions-nmr

 

 

Tools for NanoImmuno Theragnostics of Cancer

 

AREA OF KNOWLEDGE

Life Sciences Panel

 

GROUP OF DISCIPLINE

Human Biology, Microbiology, Molecular Biology, Genetics, Cell Biology, Genomics and Proteomics, Biochemistry, Basic Neuroscience

 

GROUP LEADER

Prof. José Ricardo Ramos Franco Tavares
Ricardo.franco@fct.unl.pt

 

RESEARCH PRODUCT / RESEARCH GROUP

Bionano Research Lab

https://www.ucibio.pt/research-groups/lab/bionano

 

POSITION DESCRIPTION

 

Research Project / Research Group Description

Present anti-cancer immunotherapies are successful but require more efficient and less toxic approaches. Gold nanoparticles are promising multi-modal platforms, capable of integrating multiple therapeutic (e.g., antibody and/or photothermal therapy (PTT)), diagnostic (e.g., photoacoustic imaging (PAI)) and immunomodulatory functions. Namely, gold nanostars (AuNS) are very well suited for PAI, and show high PTT potential for cancer cells, due to their high absorption coefficient in the NIR region and an optimal hydrodynamic size for intra-tumoral retention.

Sialylated glycans are frequently overexpressed in cancer microenvironment and associated with poor prognosis. Sialyl Tn (STn) may offer an opportunity for more efficacious immunotherapeutic options, as STn is only expressed in cancer, acting as immunosuppressor, impairing multiple immune cell function. Our previous studies lead us to hypothesize that targeting sialylated antigens is a strategy to overcome cancer's immunosuppression.

The MAIN AIM is to develop an immunotheragnostic approach based on bioconjugates of AuNS and sialyl Tn antigen and other cancer specific sialylated glycans. Specifically, the student will optimize the synthesis, and characterize AuNS, functionalized with specific thiolated ligands, and conjugated with the intended antibody. Characterization of the bioconjugates will be using advanced techniques such as Dynamic and Electrophoretic Light Scattering and Nanoparticle Tracking Analysis. The student will also develop relevant in vitro tests, based on immune-cancer cell co-cultures to further characterize the immunomodulatory properties of sialyl Tn. The efficacy of several antibody’s variants for antitumor capacity will be tested, and antibody blockade will be compared to sialidase treatments.

The anticancer and immunomodulatory potential of these multimodal bioconjugates will be tested in vitro. The imaging potential using PAI and the PTT potential for cancer cells will be evaluated in vivo.

 

Job position description

We are looking for self-driven and motivated PhD students. They will be integrated in a dynamic and international PhD Programme at UCIBIO-NOVA, Caparica, Portugal, with weekly Seminars and technical training for specialized equipment. They will be working in two Research Labs from the NanoImmunoTech Group: (1) the Bionano Lab where they will be involved in pursuing the main objective of the lab, namely, the development of gold nanoparticle-antibody conjugates in the context of biosensing, imaging and thermotherapy. They will use Bionanotechnology techniques (anisotropic gold nanoparticle synthesis, functionalization with active ligands, antibody-conjugation, etc.), as well as top-notch nanotechnology characterization and spectroscopic techniques, available through a scientific collaboration with CENIMAT, one of the top nanomaterials Center in the Iberian Peninsula, located in the UCIBIO-NOVA Campus; and (2) the Glycoimmunology Lab where they will be involved in pursuing the main objective of the lab, namely, the development  of immunotherapies against cancer. They will use immune techniques (immune-cancer cell co-cultures, testing antitumor capacity of antibodies variants, etc.). Antibodies will be supplied by the company CellmAbs, with close links to the Glycoimmunology Lab and also profit from a collaboration with the Chemical Glycobiology Lab from CICbioGUNE, Bilbao, Spain. Advisors will work closely with the students, for a continuous development and adjustment of their PhD Dissertation contents, with regular meetings for (re)definition of milestones and deliverables. Students will report their progress in bimonthly joint lab meetings, with all lab members involved in data discussion. Required skills for selected students are laboratory expertise in the Chemistry/Biochemistry/Biology areas. Previous experience in immunology and/or bionanotechnology is favored, but not essential, as our labs will provide and/or deepen training in those specific areas.

 

OTHER RELEVANT WEBSITES

Glycoimmunology Research Lab: https://www.ucibio.pt/research-groups/lab/glycoimmunology

CENIMAT Facilities, in the UCIBIO-NOVA Campus and available as Collaboration: https://www.cenimat.fct.unl.pt/lab-facilities

CellmAbs Biopharmaceuticals, supplying antibodies as a collaboration: https://www.cellmabs.com/

CICBioGUNE Collaborators: https://www.cicbiogune.es/people/jereno

 

 

Tracking the (intra)cellular fate and organelle trafficking of gold nanoparticles in Cancer models

 

AREA OF KNOWLEDGE

Life Sciences Panel

 

GROUP OF DISCIPLINE

Biotechnology, Bioinformatics, Pharmaceuticals, Food Technology

 

GROUP LEADER

Prof. Pedro Miguel Ribeiro Viana Baptista
pmvb@fct.unl.pt

 

RESEARCH PRODUCT / RESEARCH GROUP

The group of Nanomedicine@FCT - created and developed by Prof. Pedro V Baptista in 2003 - in the field of biofunctionalization of nanoparticles with targeting and silencing moieties and their application in drug delivery and/or gene silencing, in the assessment of toxicology of nanoparticles in in vivo models, in the development of nanobiosensors for DNA/RNA biomolecular recognition studies. At the intersection of Molecular Genetics and Nanotechnology, we have focused our research on the use of noble metal nanoparticles for new diagnostics and therapeutics platforms.

https://ucibio.pt/research-groups/lab/nanomedicine

 

POSITION DESCRIPTION

 

Research Project / Research Group Description

Nanomedicine refers to the use of nanotechnology in therapeutics and diagnostics to improve clinical outcomes, where nanoparticles play a pivotal role towards precise and efficient delivery of therapeutics to target cells. However, there is reduced information on the mechanisms and pathways involved in nanoparticle trafficking within 3D tumor spheroids (that mimic in vivo tumors), cell uptake and inside cells.

Gold nanoparticles (AuNPs) have been extensively explored in combined therapeutic concepts due to their unique properties that are well suited for the delivery of drugs to cancer cells. Still, the dynamics of nanoparticle-cell interaction is critical to understand the reciprocal effects of nanoconjugates on cells/tissues and the way these impact the former. Due to the amazing optical properties and wide range of applications, AuNPs are valuable assets to model bio-nano interactions and for the in-depth study of uptake and trafficking of AuNPs in 2D/3D cells. In fact, coupling fluorescent probes to AuNPs have recently been proposed for tracking NPs at cellular and subcellular level. These fluorescent probes (endocytic trackers) rely on modulation of fluorescence dependent on the compartments’/organelles’ pH, which correlate to internalization pathways.

We have been developing tools to trace the migration of nanoconjugates into tissues and cells since this is pivotal to the design of improved diagnostics/therapeutics nanomedicine systems. What is the fate of these nanoconjugates within cells/tissues? Can we trace these nanoplatforms through the environment and retrieve information on the environment the NPs are feeling to optimize their design? This project aims at designing and assembling NanoTracers probes that can be used to i) assess the fate of AuNPs conjugates in cells/tissues and ii) clarify the pathway NPs take from the outskirts of a 3D tumor to the core and inside cells.

 

Job position description

The project aims at the development of a AuNP probe that combines a pH dependent fluorophore (functional sensing – for sensing the intra-compartment pH), a pH independent fluorophore to act as reference (“GPS” for the nanoconjugate) and integrate the optical imaging with TEM for the critical evaluation of the sub-cellular features and organelles’ details. These tracking systems shall provide data on the reciprocal effect of cells-nanoconjugates throughout their migration into 3D spheroids and cells. In the end, we shall have developed a valuable tool and gathered critical data to assist the design of nanoconjugates for improved targeted uptake and clinical efficacy.

This project builds upon expertise of the host groups – nanoconjugates & cell models – where the suitable candidate shall be integrated. The candidate should have a strong background in biomolecular aspects of the cell (biology, biochemistry, biomedicine, biotechnology). The proposed work relates to different areas of research – from nanomedicine to cell biology.

This project will be carried out @Nanomedicine Group led by Pedro V Baptista (PVB) (supervisor) and @Human Genetics & Cancer Therapeutics Group led by Alexandra R Fernandes (ARF) (co-supervisor), both part of UCIBIO FCT-NOVA (www.ucibio.pt). Recently UCIBIO has integrated the successful application to Associated Laboratory in Bioeconomy and Health, whose strategic line focused on development of innovative (nano)therapeutics would profit from the current PhD project.

 

OTHER RELEVANT WEBSITES

This project is in collaboration with the Group of Prof. Alexandra R Fernandes – Human Genetics and cancer therapeutics Lab. The major goals of this Lab are the identification and validation of cancer diagnostics biomarkers and the development of novel combinatorial cancer therapies, using in vitro 2D (cell monocultures) and 3D (spheroids) cellular models that simulate tumor microenvironment, cancer patient´s derived tumor cells or peripheral blood, or even in vivo models.

https://ucibio.pt/research-groups/lab/human-genetics-and-cancer-therapeutics