Repositório Colecção: ICVS - conferenceAbstractsICVS - conferenceAbstractshttps://hdl.handle.net/1822/10352024-03-29T15:55:40Z2024-03-29T15:55:40ZNon-viral delivery of CRISPR/Cas9 DNA for gene editing via multivalent cationic liposome systemSousa, Diana AndradeGaspar, R.Ferreira, C. J. O.Baltazar, FátimaSilva, B. F. B.Rodrigues, L. R.https://hdl.handle.net/1822/764212022-05-26T14:42:25Z2022-03-07T09:35:16ZTítulo: Non-viral delivery of CRISPR/Cas9 DNA for gene editing via multivalent cationic liposome system
Autor: Sousa, Diana Andrade; Gaspar, R.; Ferreira, C. J. O.; Baltazar, Fátima; Silva, B. F. B.; Rodrigues, L. R.
Resumo: [Excerpt] Introduction: CRISPR/Cas9 gene editing technology has revolutionized medical research by opening new therapeutic possibilities for disease treatment such as cancer, cardiovascular, neuronal and immune disorders [1]. Even with the emergence of this technology, the lack of clinically viable delivery systems continues to hinder CRISPR therapeutic applications. Viral vectors have been sucessfully used for the delivery of CRISPR/Cas9, which whilst efficient, raise safety concerns regarding immunogenicity and insertional mutagenesis [2]. Non-viral vector such as cationic liposomes offer an attractive alternative to viruses given their low immunogenicity, high encapsulation capacity, easy synthesis and functionalization [3]. Their cationic charge mediates strong electrostatic interactions with the negative charges of nucleic acids, leading to the formation of complexes called “lipoplexes”. However, cationic lipids efficiency is still not at the level of viruses as delivery vectors. In recent years, multivalent cationic lipids have been proposed as a promising strategy to effectively deliver nucleic acids into target cells [4]. In this work, we explored the potential of multivalent cationic liposomes to deliver plasmid-based CRISPR/Cas9 systems as well as to mediate gene-editing. [...]
<b>Tipo</b>: conferenceAbstract2022-03-07T09:35:16ZExosome-mediated MEK1 silencing is a promising approach against triple negative breast cancer regressionFerreira, DéboraCátia S. PereiraCosta, MartaAfonso, JulietaLongatto, AdhemarBaltazar, FátimaKalluri, RaghuMoreira, João N.Rodrigues, L. R.https://hdl.handle.net/1822/762402022-03-01T17:13:21Z2022-03-01T17:11:41ZTítulo: Exosome-mediated MEK1 silencing is a promising approach against triple negative breast cancer regression
Autor: Ferreira, Débora; Cátia S. Pereira; Costa, Marta; Afonso, Julieta; Longatto, Adhemar; Baltazar, Fátima; Kalluri, Raghu; Moreira, João N.; Rodrigues, L. R.
Resumo: [Excerpt] Introduction: Breast cancer is a major public health problem
worldwide being the most diagnosed cancer in women [1]
. Triple negative breast cancer (TNBC) represents 10-20% of all breast cancers and is
characterized by the absence of hormone
receptors (progesterone and estrogen) and lack
of expression of epidermal growth factor receptor 2. Furthermore, it is also described to have poor
prognosis due to its propensity to metastasize to
visceral organs early in the clinical course [2–4]
.
In accordance with the lack of recurrently altered
targets at the genomic level, there is a shortage
of approved targeted agents for TNBC, remaining
cytotoxic chemotherapy the mainstay of
treatment. [...]
<b>Tipo</b>: conferenceAbstract2022-03-01T17:11:41ZElectronic cigarette aerosol: impact on embryonic lung morphologySilva-Ribeiro, TiagoCoelho, EduardoGenisheva, ZlatinaOliveira, José MariaCarvalho, PedroCorreia-Pinto, JorgeSampaio, PaulaMoura, Rute S.https://hdl.handle.net/1822/753192022-01-10T10:23:51Z2022-01-10T10:23:50ZTítulo: Electronic cigarette aerosol: impact on embryonic lung morphology
Autor: Silva-Ribeiro, Tiago; Coelho, Eduardo; Genisheva, Zlatina; Oliveira, José Maria; Carvalho, Pedro; Correia-Pinto, Jorge; Sampaio, Paula; Moura, Rute S.
Resumo: Introduction: Smoking is a major public health problem
responsible for 700000 deaths/year in Europe. Conventional
cigarettes (c-cig) exacerbates several health issues, such as
chronic obstructive pulmonary disease, fibrosis and cancer.
Tobacco use during pregnancy has serious consequences to infants,
since they become more susceptible to develop congenital
disorders, lung diseases and sudden death. Electronic cigarettes
(e-cig) have emerged as an alternative to c-cig. Previous studies
revealed that c-cig exposure impairs lung development, aggravates
wheezing and triggers inflammation. However, nothing
is known about the impact of e-cig aerosol during pulmonary
development.
Aim: Our aim was to evaluate the effect of e-cig aerosol and
c-cig smoke in the early chick embryonic lung.
Methods: Ex vivo lung explants were cultured in smoke/
aerosol medium or unexposed medium (control) for 48 hours.
Explants were assessed morphometrically. Additionally, TNF-a
levels were evaluated by ELISA.
Results: When compared to controls, c-cig treated explants
revealed a significant decrease, in all morphometric parameters,
between 15 to 30%, while e-cig treated explants displayed a
significant reduction only in lung total area and mesenchymal
perimeter (roughly 10%). Lastly, c-cig explants presented a decrease
in all morphometric parameters, between 11 to 26%,
when compared to e-cig treated explants. Additionally, e-cig and
c-cig treatment induced similar TNF-a release, that was nearly 7
times higher than control.
Conclusion: This study describes, for the first time, the impact
of e-cigs on early lung development. The results revealed that
e-cig aerosol impairs lung growth and promotes lung inflammation.
However, its impact on early lung growth seems to be less
detrimental than conventional cigarette smoke. Nevertheless, more studies are required to fully understand the effect of the aerosol in
embryo development. The validation of these effects will eventually
lead to the development of new tobacco control recommendations
to pregnant women in order to protect the fetus and childs
health.
<b>Tipo</b>: conferenceAbstract2022-01-10T10:23:50Z3-Bromopyruvate induces cytotoxicity, inhibits glycolysis and decreases migratory capacity in glioblastomas and colorectal cancer cell linesBarbosa, Ana MargaridaVieira, Joana Margarida PereiraMiranda, JoanaMoreira, RoxanaCasal, MargaridaQueirós, Odíliahttps://hdl.handle.net/1822/732582021-06-01T16:39:45Z2021-06-01T16:39:45ZTítulo: 3-Bromopyruvate induces cytotoxicity, inhibits glycolysis and decreases migratory capacity in glioblastomas and colorectal cancer cell lines
Autor: Barbosa, Ana Margarida; Vieira, Joana Margarida Pereira; Miranda, Joana; Moreira, Roxana; Casal, Margarida; Queirós, Odília
<b>Tipo</b>: conferencePoster2021-06-01T16:39:45ZMonocarboxylate transporters expression is influenced by different extracellular conditions determining the effect of 3-bromopyruvate in colorectal cancer cellsVieira, Joana Margarida PereiraAzevedo-Silva, JoãoPreto, AnaCasal, MargaridaQueirós, Odíliahttps://hdl.handle.net/1822/732572021-06-01T21:54:31Z2021-06-01T16:33:19ZTítulo: Monocarboxylate transporters expression is influenced by different extracellular conditions determining the effect of 3-bromopyruvate in colorectal cancer cells
Autor: Vieira, Joana Margarida Pereira; Azevedo-Silva, João; Preto, Ana; Casal, Margarida; Queirós, Odília
Resumo: Monocarboxylate transporters (MCTs) play a vital role in the glycolytic metabolism in cancer
cells, exporting lactate by a proton symport mechanism, maintaining intracellular pH
homeostasis and contributing for tumor microenvironment acidification and aggressiveness.
3BP is an analogous of lactate with anti-tumor properties. 3BP uptake occurs via MCTs, acting
as a glycolytic inhibitor with already identified targets, including hexokinase (HK) II and
glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Indeed, 3BP induces energy depletion
and cell death. As tumor cells overexpress MCTs, they can be used as a trap to mediate the
uptake of compounds as 3BP. However, the tumor microenvironment may influence the
expression of MCTs and consequently affect the toxic effect of 3BP. In this work, we aimed to
characterize the effect of 3BP in different colorectal cancer (CRC) cell lines by exploring 3BP
cytotoxicity and MCT1 and MCT4 expression upon different extracellular stimuli, including
extracellular pH (pHe), glucose and oxygen levels as well as short-chain fatty acids (SCFAs)
exposure. Materials and methods: Viability assays: The IC50 of 3BP was determined in basal
conditions and in the different conditions: hypoxia, starvation, different pHe and exposure to
SCFA. HCT-15 cells were incubated during 24 h with medium containing 200 µM of CoCl2
(chemical hypoxia), free-glucose medium (starvation), complete medium (w/o bicarbonate)
adjusted with HEPES buffer to pH 6.6 or 7.4 or with complete medium containing butyrate (10
mM). When cells were incubated with medium containing lactate (50 mM) or acetate (20 mM),
cells were incubated during 48 h. After that, cells were exposed to different concentrations of
3BP for 16 hour and cell viability was determined after this period of time by the SRB assay.
Metabolites quantification: HCT-15 cells were exposed to different extracellular conditions and
lactate and glucose were measured in extracellular medium. Expression assays: MCTs expression
was evaluated by Western blot assays. Protein extracts of HCT-15 cells lines, incubated in the
different conditions tested, were used. Result/Discussion: In this work, we tested the effect of
3BP in three different CRC derived cell lines: HCT15, Caco-2 and HT-29. HCT-15 cells showed to
be the most sensitive cell line to 3BP and also the one that presented the highest basal
expression of both MCT1 and CD147, a protein involved in the proper expression and activity of
MCT1 and MCT4 at cell surface. Glucose starvation and hypoxia induced an increased resistance
to 3BP in HCT-15 cells, in contrast to what happens with at acidic pHe. However, no association
with MCT1, MCT4 and CD147 expression was observed, except for glucose starvation, where a
decrease in CD147 (but not of MCT1 and MCT4) was detected. Butyrate and acetate (but not
lactate) exposure increased the expression of MCT4 (but not MCT1) and CD147. Additionally, it
was observed that the metabolic profile was affected by the 3 rd ASPIC International Congress,
10 – 11 May, Lisbon 92 different extracellular conditions. The overall results suggest that MCTs
influence 3BP effect in CRC cells, although they are not the only player in its mechanism of
action
<b>Tipo</b>: conferenceAbstract2021-06-01T16:33:19Z