Biology subjects

Cunha, J.

Publications and source records attributed to Cunha, J..

5 recordsLinked to original sources

bioRxiv2025

Multi-omics analysis of TNBC organoids identifies phosphorylation of the membrane trafficking machinery as a key event associated with FER-mediated invasion

Triple Negative Breast Cancer (TNBC) is characterised by unfavourable outcome due to the combination of its metastatic propensity, chemo-refractory behaviour and the lack of effective targeted interventions. Expression of the feline sarcoma-related (FER) kinase constitutes an independent prognostic factor that correlates with poor patient survival. FER promotes invasive behaviour in TNBC cells by regulating endosomal sorting and recycling (ESR) of adhesion proteins. Yet, the ESR machinery supporting invasion in TNBC, particularly within 3D environments, remains poorly understood. Here, we used FER-expressing TNBC patient-derived xenograft organoids (PDXOs) and MDA-MB-231 cells to identify the membrane trafficking machinery promoting invasion. Using a combination of proteomics, phospho-proteomics, and single cell RNA-sequencing, we show that the invasion of FER-expressing PDXO cells in collagen-I is mainly associated with the differential phosphorylation of membrane trafficking regulators, including SEC16A and a marked increase in Rab4-positive tubules. SEC16A depletion impairs cell invasion and reduces the number of focal adhesions and Rab4-positive tubules. Importantly, FER regulates SEC16A levels and localization, specifically in TNBC. Altogether, we identified SEC16A as a key player in FER-driven TNBC invasion, highlighting the membrane trafficking machinery as a promising target for the development of new therapeutic strategies.

cancer biology↗

bioRxiv2024

Electrospun Poly(ε-caprolactone) Membranes Modified with Heparin and Essential Fatty Acids for Biomedical Applications

The study aimed to enhance wound healing by modifying poly({varepsilon}-caprolactone) (PCL) membranes with sodium heparin (HS) and Essential Fatty Acids (EFA). Electrospinning was used to prepare the membranes containing the maximum concentration of HS and AGE, which were then sterilized with ozone. Microbiological tests confirmed effective sterilization. The membrane characterization included scanning electron microscopy (SEM) for morphological analysis, wettability studies for contact angle determination, Fourier-transform infrared spectroscopy (FT-IR) analysis, and thermogravimetry (TGA) for thermal analysis. Results indicated successful preparation and sterilization of PCL membranes modified with HS and EFA. Morphological analysis showed well-formed and randomly distributed fibers, although the PCL+HS membrane exhibited beads on its fibers. Adding HS and EFA affected the fiber diameter, with PCL+HS fibers having a smaller diameter than pure PCL and PCL+EFA fibers. Wettability analysis demonstrated modified surface properties with reduced contact angles. FT-IR analysis showed slight contributions of HS and EFA in the modified PCL membranes, while thermal analysis revealed no substantial changes in thermal stability. In conclusion, PCL membranes modified with HS and EFA can potentially accelerate wound healing, presenting an innovative and cost-effective approach to treating skin injuries.

bioengineering↗

bioRxiv2022

Grapevine leaf MALDI-MS imaging reveals the localisation of a putatively identified sucrose metabolite associated to Plasmopara viticola development

Despite well-established pathways and metabolites involved in grapevine-Plasmopara viticola interaction, information on the molecules involved in the first moments of pathogen contact with the leaf surface and their specific location is still missing. To understand and localise these molecules, we analysed grapevine leaf discs infected with P. viticola with MSI. Plant material preparation was optimised, and different matrices and solvents were tested. Our data shows that trichomes hamper matrix deposition and the ion signal. Results show that putatively identified sucrose presents a higher accumulation and a non-homogeneous distribution in the infected leaf discs in comparison with the controls. This accumulation was mainly on the veins, leading to the hypothesis that sucrose metabolism is being manipulated by the development structures of P. viticola. Up to our knowledge this is the first time that the localisation of a putatively identified sucrose metabolite was shown to be associated to P. viticola infection sites.

plant biology↗

bioRxiv2020

RNA editing in inflorescences of wild grapevine unveils association to sex and development

RNA editing challenges the central dogma of molecular biology, by modifying the genetic information at the transcription level. Recent reports, suggesting increased levels of RNA editing in plants, raised questions on the nature and dynamics of such events during development. We here report the occurrence of distinct RNA editing patterns in wild Vitis flowers during development, with twelve possible RNA editing modifications observed for the first time in plants. RNA editing events are gender and developmental stage specific, identical in subsequent years of this perennial species and with distinct nucleotide frequencies neighboring editing sites on the 5 and 3 flanks. The transcriptome dynamics unveils a new regulatory layer responsible for gender plasticity enhancement or underling dioecy evolution in Vitis.

plant biology↗

bioRxiv2020

Portuguese wild grapevine genome re-sequencing (Vitis vinifera sylvestris)

Vitis is a relevant genus worldwide. The genome of a Vitis vinifera representative (PN40024) published in 2007 boosted grapevine related studies. While this reference genome is a suitable tool for the overall studies in the field, it lacks the ability to unveil changes accumulated during V. vinifera domestication. Considering that grapevines for wine production (V. v. vinifera, hereafter vinifera) have evolved from V. v. sylvestris (hereafter sylvestris), or from a shared no-longer existing ancestor, both subspecies are quite close, but sylvestris has not been domesticated and still exist nowadays, preserving wild characteristics, making it a good material to provide insights into vinifera domestication. The difference in the reproductive strategy between both subspecies is one of the characteristics that sets them apart. While vinifera flowers are hermaphrodite with functional male and female organs, sylvestris is mostly dioecious. Male plants present flowers lacking functional carpels unable to produce grapes and female individuals have flowers with reflexed stamens producing infertile pollen but able to exhibit small and acidic grapes. In this paper, we describe the re-sequencing of the genomes from a male and a female individual of the wild sylvestris and its comparison against the reference vinifera genome.

plant biology↗