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Belder, N.

Publications and source records attributed to Belder, N..

5 recordsLinked to original sources

Torsional Force by Helical Pericytes Regulates Blood Flow in Downstream Capillaries

This study investigates the contractile properties of downstream capillaries, which are traditionally regarded as passive conduits, and addresses ongoing debates surrounding blood flow regulation. It is widely accepted that these capillaries passively dilate in response to increased blood flow in upstream microvessels, and that the helical pericytes located on them lack contractile capability, largely due to the absence of detectable -SMA expression. Challenging this prevailing view, we demonstrate that downstream capillary pericytes do express both -SMA and Myosin11, as shown using in situ hybridization on whole-mount intact retinas--unlike prior studies that relied on dissociated cells. Furthermore, Forster resonance energy transfer (FRET) analysis reveals that -SMA and Myosin11 are in sufficiently close proximity to permit actomyosin bridge cycling, a process essential for contraction. We also show that pericyte contraction can be inhibited by disrupting this molecular interaction. Distinct from the nodal constrictions caused by circular pericyte processes in upstream microvessels, we identify torsional contractions in the downstream capillaries in the retina of living mice using two-photon laser scanning microscopy (TPLSM), which regulates blood flow. These contractions provide direct evidence that downstream capillaries actively contribute to blood flow regulation. Notably, such contractions were overlooked in previous TPLSM studies that monitored only luminal diameter, unless the specialized analytical techniques we employed were applied. Our 3D modeling confirms that these torsional contractions correlate with the helical morphology of pericytes in downstream capillaries--a structure previously thought incapable of producing significant constrictive force. In conclusion, our findings provide direct evidence that downstream pericytes play an active role in regulating blood flow. They highlight a previously unrecognized mechanism--torsional contraction--that aligns with the helical structure of these pericytes and contributes to flow regulation in small-caliber capillaries located nearest to regions of high oxygen demand. TEASERDownstream Capillary Pericytes Express -SMA and Regulate Flow via Torsional Contraction

neuroscience↗

Evaluation of the Prognostic Value of Four ECM-Associated Genes in Recurrence and Metastasis of Colorectal Cancer

Colorectal cancer is a significant public health issue due to its high incidence and high mortality rate, especially when diagnosed late. Identifying new biomarkers that can predict the diagnosis and prognosis of CRC is critical to reducing the mortality rate of the disease. This study tested the relationship of four extracellular matrix genes (COL1A1, COL5A1, THBS2 and FN1) with CRC prognosis at the mRNA and protein levels. Our previous studies identified these genes as candidate prognostic biomarkers for CRC. They were analysed in two cohorts: retrospective (130 CRC tumours and 70 healthy colon tissues) and prospective (160 paired tumours and normal tissues with 180 CRC serum samples). Accordingly, in the retrospective cohort, COL1A1 (p>0.0001), COL5A1 (p>0.0001), THBS2 (0.0001) and FN1 (0.001) were expressed at higher levels in tumours compared to controls by qRT-PCR and in metastatic cases compared to non-metastatic cases. In the same cohort, immunohistochemistry analysis showed that increasing protein levels of COL1A1 and FN1 correlated with mRNA levels. In addition, high COL1A1 mRNA levels were associated with OS (p>0.011), RFS (p>0.0001), and DMFS (p>0.0001), high COL5A1 mRNA levels were associated with OS (0.042), RFS (0.013), DMFS (p>0.003) and high FN1 levels were associated with DMFS (p>0.039). In the prospective cohort, COL1A1, COL5A1 and THBS2 were found to be higher in tumour tissue compared to matched normal tissue and in metastatic tumours compared to primary tumours by qRT-PCR in 160 tumours and matched normal tissues. In this cohort, FN1 levels were unexpectedly higher in normal tissues than in tumour tissues. In the prospective cohort, where 180 CRC serum samples were also tested by ELISA, COL1A1 (p>0.0001), COL5A1 (p>0.05), and THBS2 (p>0.05) levels were higher in metastatic cases than in the sera of non-metastatic cases. As a result, this study associated COL1A1, COL5A1 and THBS2 genes, which have different data on their prognostic effects in the literature, with poor prognosis of CRC at both mRNA and protein levels.

cancer biology↗

Reduced Folate Carrier 1 (RFC1/Slc19a1) Suppression Exacerbates Blood-Brain Barrier Breakdown in Experimental Ischemic Stroke in Adult Mice

The Reduced Folate Carrier 1 (RFC1), also called solute carrier family 19 member 1 (SLC19A1/SLC19a1), is recognized for transporting folates across the blood-brain barrier (BBB). RFC1 has recently been defined as a hypoxia-immune related gene whose expression levels were induced by acute retinal ischemia, suggesting that RFC1 may have a role in the response of the brain to ischemic injury. Despite a recent human meta-analysis suggesting an association between certain RFC1 polymorphisms and the risk of silent brain infarctions, preclinical evidence concerning the potential role of RFC1 in acute ischemic stroke has yet to be presented. To investigate this, we first characterized RFC1 protein expression in mouse microvessels and pericytes which play significant roles in stroke pathophysiology. Then, we examined the temporal (1-h, 24-h, and 48-h) and spatial (infarct, periinfarct, contralateral) expression of RFC1 protein in the intraluminal transient middle cerebral artery occlusion mouse model. Finally, we knocked down RFC1 protein with RFC1-siRNA in the potential periinfarct region before induction of ischemia and investigated BBB integrity and infarct size in vivo via 7T-MRI. Moreover, we utilized a pharmacological modulation-methotrexate, a non-covalent inhibitor of RFC1- to further investigate the role of RFC1 in maintaining BBB integrity. Our study revealed that, i) RFC1 protein levels were dynamic throughout the acute phases of ischemic stroke, ii) RFC1 suppression aggravated the BBB leakage during ischemia. These results emphases the role of RFC1 in the pathophysiology of ischemic stroke and supports the evidence from human studies.

neuroscience↗

Targeting LINC00152 activates cAMP/Ca2+/ferroptosis axis and overcomes tamoxifen resistance in ER+ breast cancer

Tamoxifen has been the mainstay therapy to treat early, locally advanced, and metastatic estrogen receptor-positive (ER+) breast cancer, constituting around 75% of all cases. However, emergence of resistance is common, necessitating the identification of novel therapeutic targets. Here, we demonstrated that long-noncoding RNA LINC00152 confers tamoxifen resistance via blocking tamoxifen-induced ferroptosis, an iron-mediated cell death. Mechanistically, inhibiting LINC00152 reduces the mRNA stability of phosphodiesterase 4D (PDE4D), leading to activation of cAMP/PKA/CREB axis and increased expression of TRPC1 Ca2+ channel. This causes cytosolic Ca2+ overload and generation of reactive oxygen species (ROS) that is, on one hand, accompanied by downregulation of FTH1, a member of the iron sequestration unit, thus increasing intracellular Fe2+ levels; and on the other hand, inhibition of the peroxidase activity upon reduced GPX4 and xCT levels. These ultimately induce lipid peroxidation and ferroptotic cell death in combination with tamoxifen. Overexpressing PDE4D rescues LINC00152 inhibition-mediated tamoxifen sensitization by de-activating the cAMP/Ca2+/ferroptosis axis. Importantly, high LINC00152 expression is significantly correlated with high PDE4D/low ferroptosis and worse survival in multiple cohorts of tamoxifen- or tamoxifen-containing endocrine therapy-treated ER+ breast cancer patients. Overall, we identified LINC00152 inhibition as a novel mechanism of ferroptosis induction and tamoxifen sensitization, thereby revealing LINC00152 and its effectors as actionable therapeutic targets to improve clinical outcome in refractory ER+ breast cancer.

cancer biology↗

Reduced Folate Carrier 1 is Present in Retinal Microvessels and Contributes to the Regulation of The Inner Blood Retinal Barrier in Health and Retinal Ischemia

BackgroundReduced folate carrier 1 (RFC1; SLC19a1) is the main responsible transporter for the B9 family of vitamins named folates, which are essential for normal tissue growth and development. While folate deficiency resulted in retinal vasculopathy, the expression and the role of RFC1 in blood-retinal barrier (BRB) are not well known. MethodsWe used whole mount retinas and trypsin digested microvessel samples of adult mice. To knockdown RFC1, we delivered RFC1-targeted short interfering RNA (RFC1-siRNA) intravitreally; while, to upregulate RFC1 we delivered lentiviral vector overexpressing RFC1. Retinal ischemia was induced 1-hour by applying FeCl3 to central retinal artery. We used RT-qPCR and Western blotting to determine RFC1. Endothelium (CD31), pericytes (PDGFR-beta, CD13, NG2), tight-junctions (Occludin, Claudin-5 and ZO-1), main basal membrane protein (Collagen-4), endogenous IgG and RFC1 were determined immunohistochemically. ResultsOur analyses on whole mount retinas and trypsin digested microvessel samples of adult mice revealed the presence of RFC1 in the inner BRB and colocalization with endothelial cells and pericytes. Knocking down RFC1 expression via siRNA delivery resulted in the disintegration of tight junction proteins and collagen-4 in twenty-four hours, which was accompanied by significant endogenous IgG extravasation. This indicated the impairment of BRB integrity after an abrupt RFC1 decrease. Furthermore, lentiviral vector-mediated RFC1 overexpression resulted in increased tight junction proteins and collagen-4, confirming the structural role of RFC1 in the inner BRB. Acute retinal ischemia decreased collagen-4 and occludin levels and led to an increase in RFC1. Besides, the pre-ischemic overexpression of RFC1 partially rescued collagen-4 and occludin levels which would be decreased after ischemia. ConclusionIn conclusion, our study clarifies the presence of RFC1 protein in the inner BRB, which has recently been defined as hypoxia-immune-related gene in other tissues and offers a novel perspective of retinal RFC1. Hence, other than being a folate carrier, RFC1 is an acute regulator of the inner BRB in healthy and ischemic retinas.

neuroscience↗