Biology subjects

Somoza, A.

Publications and source records attributed to Somoza, A..

4 recordsLinked to original sources

bioRxiv2026

Carboxymethyl Dextran Coated Iron Oxide Magnetic Nanoparticles as Vehicles for CRISPR Ribonucleoproteins

The efficient and safe intracellular delivery of CRISPR ribonucleoproteins (RNPs) remains a major challenge for the development of genome editing strategies. Magnetic iron oxide nanoparticles (MNPs) offer an attractive and biocompatible delivery platform. However, their use for the delivery of functional CRISPR RNPs remains comparatively underexplored. Here, we developed carboxymethyl dextran (CMD)-coated MNPs as non-viral carriers for Cas9 and Cas12a RNPs. Nanocomplexes were formed through electrostatic interactions, without the need for chemical coupling reagents or additional surface functionalization. Both RNPs efficiently associated with MNPs, and the resulting nanocomplexes exhibited stable physicochemical properties and retained their cargo in serum-supplemented culture medium for up to 24 hours. Cellular uptake was confirmed by microscopy, while biocompatibility assays revealed no overt cytotoxicity. Functional genome editing was demonstrated at reporter and endogenous gene loci in HEK-293T cells, as well as in primary human dermal fibroblasts. Finally, we explored the therapeutic potential of this platform for recessive dystrophic epidermolysis bullosa (RDEB) by targeting COL7A1 exon 80, with the aim of removing the prevalent c.6527insC mutation while preserving the reading frame. Cas9 RNP nanocomplexes enabled exon 80 deletion. These findings establish CMD-coated MNPs as a versatile platform for the delivery of Cas9 and Cas12a RNPs and support further investigation of magnetic nanoparticle-mediated genome editing for inherited skin disorders.

molecular biology↗

bioRxiv2025

Hairpin-Functionalized Gold Nanoparticles as an Adaptable Platform for Detecting MicroRNA Signatures

Early, accurate, and fast diagnosis is essential to ensure positive health outcomes, through effective treatment interventions and disease control, as well as the study of physiological changes related to gene expression. Liquid biopsies and point-of-care (PoC) detection are valuable tools to achieve this goal, allowing the study and monitoring of a patients molecular profile in a timely and simple manner. MicroRNAs (or miRNAs) have been proposed as biomarkers for detection in liquid biopsies, as they are stable in body fluids and their dysregulation is associated with many diseases. In this work, a sensor based on gold nanoparticles (AuNPs) functionalized with oligonucleotides is reported, aiming for the detection of nucleic acids, in particular miRNAs. This sensor is based on the recognition of target sequences by hairpin-shaped oligonucleotide probes, which allows the modulation of the colloidal stability of the AuNPs, producing color changes detectable with the naked eye. The system is used to detect a panel of miRNAs, demonstrating its versatility for the detection of relevant nucleic acid signatures. The sensor detects single miRNAs with good sensitivity and selectivity and, what is more, it can be used to recognize several miRNAs simultaneously at picomolar concentrations. The system was further adapted to a lateral flow assay (LFA) format, producing a visible colored line on lateral flow test strips, and coupled with isothermal amplification to reach femtomolar detection levels. Its properties make it suitable for use in the point-of-care (PoC), contributing to fast and early detection of pathological and physiological molecular profiles.

molecular biology↗

bioRxiv2025

Targeting of ibrutinib resistance driving pathways by miR-28 in ABC-DLBCL

Diffuse large B-cell lymphoma (DLBCL) is the most common aggressive B-cell lymphoma. Although many patients respond well to R-CHOP immunochemotherapy, those with the activated B-cell (ABC) subtype are often refractory or relapse. Bruton tyrosine kinase (BTK) inhibitors such as ibrutinib have improved outcomes, but acquired resistance limits their long-term efficacy. Here, we modeled the development of ibrutinib resistance in ABC-DLBCL and investigated whether the BCR-signaling regulator microRNA-28 (miR-28) can block this process. Using flow cytometry-based competition assays, multicolor clonal barcoding, transcriptomic profiling, and xenograft models, we found that miR-28 expression impairs the emergence of ibrutinib-resistant ABC-DLBCL cells. Mechanistically, miR-28 interferes with the clonal selection process triggered by ibrutinib treatment and rewires transcriptional programs by downregulating mitochondrial and mTOR signaling pathways critical for resistance development. Furthermore, the miR-28-repressed gene signature associated with ibrutinib resistance correlates with improved survival in ibrutinib-treated patients from the PHOENIX trial cohort with the MCD genetic subtype, which is associated with ABC-DLBCL. Finally, the targeted therapeutic delivery of miR-28 via aptamer-guided nanoparticles suppresses ibrutinib-resistant tumor growth in vivo. These findings identify miR-28 as an effective inhibitor of ibrutinib resistance, underscoring its translational potential as an adjunct strategy in ABC-DLBCL therapy. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=188 SRC="FIGDIR/small/687947v2_ufig1.gif" ALT="Figure 1"> View larger version (70K): org.highwire.dtl.DTLVardef@8e5913org.highwire.dtl.DTLVardef@17a19c3org.highwire.dtl.DTLVardef@13178f2org.highwire.dtl.DTLVardef@fb5d9b_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2022

Natural killer (NK) cell-derived extracellular-vesicle shuttled microRNAs control T cell responses

Natural killer (NK) cells recognise and kill target cells undergoing different types of stress. NK cells are also capable of modulating immune responses. In particular, they regulate T cell functions. Small RNA next-generation sequencing of resting and activated NK cells and their secreted EVs led to the identification of a specific repertoire of NK-EV-associated microRNAs and their post-transcriptional modifications signature. Several microRNAs of NK-EVs, namely miR-10b-5p, miR-92a-3p and miR-155-5p, specifically target molecules involved in Th1 responses. NK-EVs promote the downregulation of GATA-3 mRNA in CD4+ T cells and subsequent T-bet de-repression that leads to Th1 polarization and IFN-{gamma} and IL-2 production. NK-EVs also have an effect on monocyte and moDCs function, driving their activation and increased presentation and co-stimulatory functions. Nanoparticle-delivered NK-EV microRNAs partially recapitulate NK-EV effects in vivo. Our results provide new insights on the immunomodulatory roles of NK-EVs that may help to improve their use as immunotherapeutic tools.

immunology↗