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Joris, V.

Publications and source records attributed to Joris, V..

5 recordsLinked to original sources

Posterior insula oscillations are not modulated by changes in perceived intensity: Evidence from human intracerebral EEG

The human insular cortex is involved in a multitude of processes essential for survival, but its precise role in pain perception remains debated. While it has been shown that ongoing oscillatory activity in the posterior insula is preferentially modulated by thermonociceptive input, it is unclear whether these modulations are functionally related to changes in pain perception. To assess this link, neural responses to sustained periodic thermonociceptive and non-nociceptive vibrotactile stimulation delivered at a frequency of 0.2 Hz were measured using intracerebral electrode contacts located in the anterior (n=63) and posterior (n=37) insular cortices of 10 patients undergoing presurgical evaluation for focal epilepsy. An arithmetic task was employed concomitant to the stimulation, aiming to reduce participants perceived intensity while leaving the physical stimulation parameters unchanged. A frequency-tagging analysis approach was used to assess the modulation of the ongoing neural oscillations. As expected, the perception of the stimulation was significantly reduced during the arithmetic task to a similar extent in both modalities. Yet, no congruent reduction in the frequency-tagged oscillatory responses was found in the posterior insula during distraction, for either of the modalities. On the contrary, during distraction, vibrotactile stimulation elicited larger responses in the anterior insula. These findings suggest that top-down modulation via distraction differentially affects anterior insula responses to innocuous vibrotactile input, while thermonociceptive processing appears to involve additional neural systems during distraction that are not equivalently engaged by vibrotactile stimulation.

neuroscience↗

MiR-125b differentially impacts mineralization in dexamethasone and calcium-treated human mesenchymal stem cells

Bone metabolism is highly regulated, and microRNAs (miR) can contribute to this process. Among them, miR-125b is well-known to enhance osteoporosis and reduce osteogenic differentiation of human mesenchymal stem cells (hMSCs). In this work we aim to evaluate and understand how miR-125b modulates mineralization of hMSCs in two different in vitro models. Cells were cultured in dexamethasone or calcium medium and transfected with miR-125b mimic. Exposure to dexamethasone or calcium medium increased mineralization of hMSCs and was associated with decreased miR-125b expression. Transfection of miR-125b mimic in dexamethasone-treated cells increased mineralization while it decreased it in calcium-treated cells. Levels of osteogenic markers presented the same difference. We identified STAT3, p53 and RUNX2 as direct targets of miR-125b in hMSCs. While these targets remained identical in both treatments, their modulation after transfection was different. We showed that miR-125b mimicking differentially modulated the expression of the miR-199a/214 cluster, probably via STAT3/miR-199a/214, and p53/miR-214 pathways. In conclusion, miR-125b affinity for targets implicated in bone remodeling changed depending on the in vitro models used to induce mineralization and led to opposite physiological effect. This works shows the complexity of drugs such as dexamethasone and opens the door for new in vitro models of mineralization.

molecular biology↗

FGF9 treatment reduces off-target chondrocytes from iPSC-derived kidney organoids

Renal failure due to drug nephrotoxicity or disease is frequently observed in patients. The development of in vitro models able to recapitulate kidney biology offers new possibilities to study drug toxicity or model diseases. Induced pluripotent stem cell-derived kidney organoids already show promise, but several drawbacks must be overcome to maintain them in culture, among which is the presence of non-renal cell populations such as cartilage. We modified the culture protocol and maintained kidney organoids in medium containing FGF9 for one additional week compared to the control protocol (Takasato). In comparison to the control, the FGF9-treated kidney organoids had reduced cartilage at day 7+25 and diminished chondrocyte marker expression. Importantly, the renal structures assessed by immunofluorescence were unaffected by the FGF9 treatment. This reduction of cartilage produces a higher quality kidney organoid that can be maintained longer in culture to improve their maturation for further in vivo work. HighlightsO_LIKidney organoids develop cartilage between days 7+18 and 7+25 C_LIO_LIExtending the FGF9 supplementation reduces cartilage and chondrocyte markers C_LIO_LIFGF9-treated organoids present less EMT marker expression than control organoids C_LIO_LIRenal structures are not affected by the extended FGF9 treatment C_LI eTOC blurbLaPointe and colleagues showed that a one-week extension of FGF9 supplementation in iPSC-derived kidney organoids leads to a reduction of off-target cartilage. The renal structures are not impacted by FGF9 treatment and EMT markers are reduced. This modified organoid protocol will enable longer culture periods, a benefit for the use of organoids for screening or therapies.

cell biology↗

Ultrastructural comparison of human kidney organoids and human fetal kidneys reveals features of hyperglycemic culture

Induced pluripotent stem cell (iPSC)-derived kidney organoids have the potential for a large variety of applications. However, they do not persist long in culture, for which reasons are still unclear. Furthermore, their morphological maturation, an essential feature for kidney function, has not been sufficiently assessed. Kidney organoids are transcriptionally much alike end-of-first-trimester fetal kidneys and present many of the same cell types. From large transmission electron microscopy tilescans of specific regions of interest, we compared the ultrastructures of iPSC-derived kidney organoids at various timepoints to human fetal kidneys of the first trimester. Unlike healthy fetal kidneys, large glycogen deposits developed over time in all organoid cell types, but particularly in podocytes and in chondrocytes, one of the off-target populations that contaminate the culture. Deeper investigation showed that glycogen synthase kinase 3b (GSK3{beta}) levels and activation were diminished over time, correlated with the accumulation of glycogen. Activated YAP was strongly expressed and large lipid droplets accumulated over time in proximal tubules. Accordingly, EGFR signaling increased significantly over time. Mitochondria in glomeruli and tubules contained few or no cristae, indicating mitochondrial damage. Together these features are known for hyperglycemic cultures and diabetic nephropathy. Measuring the glucose concentration in the kidney organoid culture medium showed a concentration of 2.94 g/mL, which is considered an elevated, pre-diabetic-like concentration in vitro. In summary, our ultrastructural assessment of iPSC-derived kidney organoids using an age-matched fetal human reference allowed the evaluation of cellular morphology, and we identified intracellular features that can inform the cellular state, which is particularly important while physiological testing of organoids is limited. Translational StatementKidney organoids hold promise as a future treatment for patients with end-stage kidney disease. The engineering of kidney organoids with correct and healthy morphology in vitro is therefore essential, to guarantee functionality after transplantation. The present study provided deeper insights into the structural organization and ultrastructure of cells in kidney organoids compared to age-matched human fetal kidneys. Accordingly, we found several features in the regular kidney organoid culture, which are known to occur in hyperglycemic cultures and diabetic nephropathy, indicating that the current medium composition may be inducing pathological cellular phenotypes. This study therefore creates a better understanding of current limitations in the kidney organoid culture, increases knowledge of their function and cellular organization, and sets the foundation for further research to create advanced organoids.

bioengineering↗

Enhanced microvasculature formation and patterning in iPSC-derived kidney organoids cultured in physiological hypoxia

Functional kidney organoids have the potential to be used in implantable kidney grafts for patients with end-stage kidney disease, because they have been shown to self-organize from induced pluripotent stem cells into most important renal structures. To date, however, long-term kidney organoid culture has not succeeded, as nephrons lose their phenotype after approximately 25 days. Furthermore, the renal structures remain immature with diminishing endothelial networks with low connectivity and limited organoid invasion. We hypothesized that introducing long-term culture at physiological hypoxia, rather than the normally applied non-physiological, hyperoxic 21% O2, could initiate angiogenesis, lead to enhanced growth factor expression and improve the endothelial patterning. We therefore cultured the kidney organoids at 7% O2 instead of 21% O2 for up to 25 days and evaluated nephrogenesis, VEGF-A expression and vascularization. Whole mount imaging revealed a homogenous morphology of the endothelial network with enhanced sprouting and interconnectivity when the kidney organoids were cultured in hypoxia. Three-dimensional quantification confirmed that the hypoxic culture led to an increased average vessel length, likely due to the observed upregulation of proangiogenic VEGF-A189 mRNA and downregulation of the antiangiogenic protein VEGF-A165b measured in hypoxia. This research indicates the importance of optimization of oxygen availability in organoid systems and the potential of hypoxic culture conditions in improving the vascularization of organoids. Significance statementCulturing kidney organoids in a hypoxic environment improved patterning of the endothelial network and improved vascularization. These findings may help improve the quality of kidney organoids, and could eventually improve the kidney graft for transplantation in patients with end-stage kidney disease. Furthermore, the organoids will be more suitable for drug testing and in developmental biology. The findings might also be translatable to other organoid models containing endothelial cells. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=199 SRC="FIGDIR/small/473849v1_ufig1.gif" ALT="Figure 1"> View larger version (61K): org.highwire.dtl.DTLVardef@d5cde7org.highwire.dtl.DTLVardef@c0ff46org.highwire.dtl.DTLVardef@b76191org.highwire.dtl.DTLVardef@166823d_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗