Biology subjects

Rico, P.

Publications and source records attributed to Rico, P..

6 recordsLinked to original sources

bioRxiv2025

NaBC1 Acts as a Mechanosensitive Co-regulator of Fibronectin-binding Integrins Adhesion and Myoblast Polarization

Cell-matrix interactions are central to the regulation of cell mechanics, polarity, and signal transduction. In this study, we investigate the contribution of the borate transporter NaBC1 to myoblast adhesion dynamics and mechanotransductive responses. Using C2C12 myoblasts cultured on fibronectin-coated substrates, we show that NaBC1 activation rapidly reinforces cell-substrate attachment, leading to accelerated spreading and the establishment of polarized cell morphologies. These changes are associated with the assembly of enlarged focal adhesions and a marked slowdown of actin retrograde flow, consistent with increased force transmission across adhesion sites. NaBC1 stimulation also induces a coordinated increase in the expression of fibronectin-binding integrins, including 5{beta}1 and v{beta}3, at both transcriptional and protein levels. Proximity ligation assays reveal an enhanced spatial association between NaBC1 and these integrins at the cell membrane, supporting the formation of cooperative adhesion complexes. In parallel, fluorescently labelled boron accumulates at focal adhesions and within intracellular compartments such as mitochondria, lysosomes, and the endoplasmic reticulum, suggesting a link between NaBC1-dependent adhesion signaling and subcellular organization. Importantly, the adhesive and mechanotransductive effects driven by NaBC1 are strictly dependent on fibronectin integrity and are lost on mutant fibronectins lacking RGD or synergy motifs, as well as on laminin-111 substrates that do not support molecular clutch engagement. Together, these findings identify NaBC1 as an integral component of fibronectin-integrin adhesion systems, contributing to the regulation of myoblast mechanics and polarity through cooperative interactions with fibronectin-binding integrins.

bioengineering↗

bioRxiv2024

NaBC1 boron transporter enables myoblast response to substrate rigidity via fibronectin-binding integrins

Cells are sensitive to the physical properties of their microenvironment and transduce them into biochemical cues that trigger gene expression and alter cell behavior. Numerous proteins, including integrins, are involved in these mechanotransductive events. Here, we identify a novel role for the boron transporter NaBC1 as a mechanotransducer. We demonstrate that soluble boron ions activate NaBC1 to enhance cell adhesion and intracellular tension in C2C12 myoblasts seeded on fibronectin-functionalised polyacrylamide (PAAm) hydrogels. Retrograde actin flow and traction forces exerted by these cells are significantly increased in vitro in response to both increased boron concentration and hydrogel stiffness. These effects are fibronectin and NaBC1-mediated as they are abrogated in hydrogels coated with laminin-111 in place of fibronectin and in esiRNA NaBC1-silenced cells. Our findings thus demonstrate that NaBC1 controls boron homeostasis and also functions as a mechanosensor.

bioengineering↗

bioRxiv2023

Injectable borax-loaded alginate hydrogels reduce muscle atrophy, inflammation and generate neuroprotection in the SOD1G93A mouse model of ALS

Amyotrophic Lateral Sclerosis (ALS) is the most frequent and fatal condition that causes motor neuron loss and skeletal muscle paralysis. Although ALS is associated with mutations in over 40 genes, its etiology remains largely elusive without a cure or effective treatment. Historically considered the prototype of motor neuron diseases, ALS is defined today as a multisystem disorder that presents several changes in non-neuronal cell types, such as pathological changes in muscle occurring before disease onset and independent from motor neuron degeneration (dying back hypothesis). We base on the hypothesis that skeletal muscle may have an active contribution to disease pathology and thus we consider skeletal muscle tissue as a therapeutic target for ALS. In previous works, we have demonstrated that boron transporter NaBC1 (encoded by the SLC4A11 gene), after activation co-localizes with integrins and growth factor receptors producing a functional cluster that synergistically enhances crosstalk mechanisms accelerating muscle repair. In this work, we aimed to study the effects of borax (B) in a SOD1 mouse model of ALS targeting muscle. We have engineered and characterized injectable alginate-based hydrogels with controlled local borax release to effectively activate muscle NaBC1 in vivo. Treated mice presented improved motor function and extended survival correlated with the activation of essential muscle metabolic pathways, resulting in an enhanced muscle repair response and reduced muscle atrophy and inflammation. Interestingly, the activation of muscle repair mechanisms at the local level produced retrograde neuroprotection by motor neuron preservation and reduction in neuroinflammation. Altogether, this work presents evidence supporting the involvement of muscle tissue in ALS pathology, reinforcing skeletal muscle as a primary target to develop new therapies for ALS. We propose a novel strategy based on NaBC1 activation for ALS muscle regeneration. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=139 SRC="FIGDIR/small/567052v2_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@16ba3e7org.highwire.dtl.DTLVardef@1d5c8c3org.highwire.dtl.DTLVardef@6f8775org.highwire.dtl.DTLVardef@2623fc_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

bioRxiv2020

Lithium directs embryonic stem cell differentiation versus hemogenic endothelium

The discovery that the definitive hematopoietic stem cells (HSCs) derive from specialized regions of the endothelium, known as the hemogenic endothelium (HE), shed a good deal of light on HSC embryonic developmental processes. This knowledge opened up new possibilities for the design of new strategies to obtain HSCs in vitro from pluripotent stem cells (PSCs). Previous advances in this field have shown that the Wnt/{beta}-catenin signaling pathway plays a key role in PSC-derived HSC formation. In this work, we identified lithium, a GSK3 inhibitor, as an element capable of stabilizing {beta}-catenin and inducing ESC differentiation in the mesoderm lineage and subsequently in the HE, highly consistent with the role of Wnt agonists on ESC differentiation. ESCs treated with 10 mM lithium express CD31+, Sca-1+, Nkx2-5+ and Runx1+ cells characteristic of HE cells. The ability of lithium-treated ESCs to further derive into HSCs was confirmed after defined maturation, resulting in rounded cell aggregates positive for fetal and mature HSCs markers, confirming the endothelial to hematopoietic transition. Our results represent a novel strategy for generating HSC in vitro as a multipotent source of stem cells for blood and muscle disease therapies.

cell biology↗

bioRxiv2020

Boron induces osteogenesis by stimulating NaBC1 in cooperation with BMPR1A

The intrinsic properties of Mesenchymal Stem Cells (MSCs) make them ideal candidates for tissue engineering applications as they are regulated by the different signals present in the stem cell niche. Considerable efforts have been made to control stem cell behavior by designing material system approaches to engineer synthetic extracellular matrices and/or include soluble factors in the media. This work proposes a novel and simple approach based on ion-channel stimulation to determine stem cell fate that avoids the use of growth factors (GFs). We used boron ion - essential item in cell metabolism - transported inside cells by the NaBC1-channel. Addition of boron alone enhanced MSC adhesion and contractility, promoted osteogenesis and inhibited adipogenesis. The stimulated NaBC1 promoted osteogenesis via activation of the BMP canonical pathway (comprising Smad1 and YAP nucleus translocation and osteopontin expression) through a mechanism that involves simultaneous NaBC1/BMPR1A and NaBC1/5{beta}1/v{beta}3 co-localization. We describe a novel function for NaBC1 as a mechanosensitive ion-channel capable of interacting and stimulating GF receptors and fibronectin-binding integrins. Our results open up new biomaterial engineering approaches for biomedical applications by a cost-effective strategy that avoids the use of soluble GFs.

cell biology↗

bioRxiv2019

Engineered full-length Fibronectin-based hydrogels sequester and present growth factors to promote regenerative responses in vitro and in vivo

Extracellular matrix (ECM)-derived matrices such as Matrigel are used to culture numerous cell types in vitro and can recapitulate certain ECM functions that support cell growth and differentiation. However, ECM-derived matrices suffer lot-to-lot variability, undefined composition and lack of controlled physical properties. There is a need to develop rationally designed synthetic matrices that can also recapitulate ECM roles. Synthetic matrices have certain limitations as they use synthetic peptides or fragments whereas the ECM consists of full proteins. Here, we report the development of degradable, PEG-based hydrogels of controlled stiffness that incorporate full-length fibronectin (FN) to enable solid-phase presentation of growth factors in a physiological manner. We demonstrate, in vitro and in vivo, the effect of incorporating vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP2), in these hydrogels to enhance angiogenesis and bone regeneration, respectively. We show that the solid-state presentation of growth factors enables very low growth factor doses to achieve regenerative effects.

bioengineering↗