Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Patient-derived Enteroids provide a Platform for the Development of Therapeutic Approaches in Microvillus Inclusion Disease

ABSTRACT: Microvillus Inclusion Disease (MVID), caused by loss-of-function mutations in the motor protein Myosin Vb (MYO5B), is a severe infantile disease characterized by diarrhea, malabsorption, and acid-base instability, requiring intensive parenteral support for nutritional and fluid management. Human patient-derived enteroids represent a model for investigation of monogenic epithelial disorders but are a rare resource from MVID patients. We developed human enteroids with different loss-of function MYO5B variants and showed that they recapitulated the structural changes found in native MVID enterocytes. Multiplex Immunofluorescence imaging of patient duodenal tissues revealed patient-specific changes in localization of brush border transporters. Functional analysis of electrolyte transport revealed profound loss of Na+/H+ exchange (NHE) activity in MVID patient enteroids with near-normal chloride secretion. The chloride channel-blocking anti-diarrheal drug, Crofelemer, dose-dependently inhibited agonist-mediated fluid secretion. MVID enteroids exhibited altered differentiation and maturation versus healthy enteroids. Gamma-secretase inhibition with DAPT recovered apical brush border structure and functional Na+/H+ exchange activity in MVID enteroids. Transcriptomic analysis revealed potential pathways involved in the rescue of MVID cells including serum- and glucocorticoid-induced protein kinase 2 (SGK2), and NHE regulatory factor 3 (NHERF3). These results demonstrate the utility of patient-derived enteroids for developing therapeutic approaches to MVID.

ORGANISM(S): Homo sapiens

PROVIDER: GSE240928 | GEO | 2023/08/16

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Similar Datasets

Lysophosphatidic acid treatment improves brush border maturation and apical SGLT1 activity in Myo5b deficient mice, a model of MVID

Project description:Myosin Vb (Myo5b) is an essential trafficking protein for membrane recycling in gastrointestinal (GI) epithelial cells and its inactivating mutation causes the congenital diarrheal disease, microvillus inclusion disease (MVID). We previously reported that Myo5b deficiency in mice causes mislocalization of SGLT1 and NHE3, but retained apical function of CFTR, resulting in malabsorption and secretory diarrhea. Activation of lysophosphatidic acid (LPA) receptors can improve diarrhea, but the effects of LPA on MVID symptoms is unclear. We therefore tested whether LPA treatment can ameliorate the epithelial deficits in Myo5b knockout (KO) mice. Methods: Adult tamoxifen-induced, intestine-specific, Myo5b KO (VillinCreERT2;Myo5bflox/flox) and littermate control mice were treated with LPA, an LPAR2 agonist (GRI977143), or vehicle for 4 days after a single tamoxifen injection. Apical SGLT1 and CFTR activities were measured in Üssing chambers. The localization of membrane transporters was evaluated by immunostaining in mouse tissues and enteroids. RNA sequencing and enrichment analysis were performed with isolated jejunal epithelial cells. Results: Daily treatment with LPA decreased the frequency of multivesicular bodies and the expression of cathepsins, but did not affect the formation of microvillus inclusions in Myo5b KO mice. LPA partially restored the brush border height and the localization of SGLT1 and NHE3 in Myo5b KO small intestine and enteroids. SGLT1-dependent short-circuit current (Isc) was increased and abnormal CFTR activities were suppressed in LPA-treated jejunum compared to that of vehicle-treated Myo5b KO mice. Conclusions: Cell autonomous LPA signaling may modulate a Myo5b-independent trafficking mechanism and the brush border maturation, demonstrating a therapeutic potential for LPA in the treatment of MVID.

2019-10-24 | GSE139302 | GEO

Coordinated assembly of NHE3, IRBIT, NHERF1, and ezrin ameliorates fluid malabsorption in diabetes

Project description:Clinical evidence supports the occurrence of intermittent diarrhea in many type 1 diabetes mellitus (T1DM) patients. Others and we found that net fluid absorption rate is dramatically lower in the ileum of T1DM murine models. However, the identity of molecules that contribute to fluid malabsorption in the gut remains unknown. Considering the importance of ion transporters and channels for intestinal fluid absorption, we reasoned that their expression level at the luminal membrane is altered under diabetic conditions. To this end, we analyzed the brush border membrane vesicles (BBMVs) from the ileum of control and DM mice by proteomic analysis. The expression levels of Cl-/HCO3- exchanger SLC26A3/DRA and cystic fibrosis transmembrane conductance regulator (CFTR) were not significantly different between control and DM mice. Cl-/HCO3- exchanger Slc26a6/PAT1 and Na+/H+ exchanger 3 (NHE3) were not detected. Interestingly, cytoskeleton scaffold proteins that regulate NHE3, including NHERF1-3 and ezrin, were all significantly lower in diabetic animals.

2016-04-27 | PXD001859 | Pride

Human Renal Tubular Anti-Brush Border Antigen (ABBA) Discovery

Project description:Discovery of target antigen resulting in autoimmune form of renal interstitial nephritis using a brush border biotinylation, affinity purification of target antigen using patient sera, and characterization of biotinylated species in the immunopurified complex. Results were confirmed by IF of patient biopsies and IB using recombinant target antigen.

2017-04-04 | MSV000080885 | MassIVE

Loss-of-function of MYO5B induces epithelial cell scattering in enterocytes

Project description:A non-functional myosin Vb motor in duodenal enterocytes results in disruption of epithelial cell polarity characterized by complete loss of microvilli and mislocalization of apical brush border proteins in the cytoplasm which finally cause a devastating disease in neonates with severe malabsorption defects accompanied by protracted diarrhea during infancy, classified as microvillus inclusion disease (MVID). The exact mechanisms how loss-of-function of MYO5B induces polarity loss are not completely understood in MVID pathogenesis. Obtaining better insights in cell polarity defects caused by loss of MYO5B, we performed microarray- in combination with protein expression-analysis in an inducible CaCo2 MYO5B RNAi cell system. Surprisingly, in MYO5B-depleted CaCo2 cells, CDH1 coding for the cell adhesion protein E-Cadherin and important for cell adhesion and therefore maintenance of cell polarity, was significantly downregulated. Interestingly, mesenchymal cell markers, specifically Vimentin and N-Cadherin, physiologically not expressed in differentiated epithelium, were upregulated and accompanied by increased phospho-c-jun levels in the nucleus. Importantly phospho-c-jun was also found in nuclei of duodenal enterocytes in MVID patients, indicating loss of MYO5B induces epithelial cell scattering in enterocytes. 3 Myosin 5B KD versus 3 control samples

2013-03-14 | E-GEOD-45134 | biostudies-arrayexpress

Cubilin is a Novel Target Antigen in Anti-Brush Border Antibody Disease

Project description:Autoimmune diseases that affect the kidney proximal tubule (PT) cause proteinuria and are implicated in the pathogenesis of kidney failure. Previous evidence demonstrated that circulating anti-brush border antibodies (ABBA) target megalin (LRP2) but whether other components of the PT drive autoimmunity was unknown. Here we investigate a patient with LRP2-negative ABBA disease, and applied immunoprecipitation, mass spectrometry, and confocal imaging to identify cubilin as a novel antigen associated with the autoimmune disorder.

2023-04-06 | PXD030159 | Pride

Elucidation of molecular functions of the protocadherin CDHR5 in intestinal epithelial cells

Project description:Intestinal epithelial cells are covered by the brush border, which consists of densely packed microvilli. The Intermicrovillar Adhesion Complex (IMAC) connects the microvilli and is required for proper organization of the brush border. The protocadherin CDHR5 is an important member of the IMAC, but it is unclear whether CDHR5 has mainly structural functions or is also involved in cellular signaling. This issue was resolved with a CDHR5 knockout mouse model. Intestinal epithelial cells were isolated from the small intestine of four wild-type and four CDHR5 knockout mice. Bulk RNA sequencing was performed to provide insight into the functions of CDHR5 in cellular signaling and gene regulation.

2023-04-13 | E-MTAB-12525 | biostudies-arrayexpress

Transcriptional profiles of yolk sac upon HNF4A loss in mice by RNA-seq

Project description:Knockout of HNF4A in yolk sac resulted in decreased expression levels of brush border genes and increased expression levels of stress fiber genes.

2021-03-15 | GSE166125 | GEO

Pulmonary Neuroendocrine Cells Regulate Gas Exchange Through Controlling Lung Fluid Balance

Project description:Limiting fluid in lung is critical for efficient gas exchange. Here we discovered a mechanism of neuropeptidergic control of lung fluid balance by pulmonary neuroendocrine cells (PNECs), potent sensors of chemical and mechanical cues. We studied the first animal model of neuroendocrine cell hyperplasia of infancy (NEHI), which faithfully recapitulated patient phenotypes including PNEC hyperplasia and impaired gas exchange. Double mutants showed that increased PNECs and excess PNEC products such as CGRP are responsible for poor gas exchange, acting through downregulating endothelial junctions, increasing vessel leakage and fluid accumulation. Endothelium-specific inactivation of CGRP receptor, or treatment with CGRP receptor antagonist reduced fluid and improved gas exchange. In lungs with acute respiratory distress syndrome (ARDS), including those caused by COVID-19, there was a striking increase of CGRP-expressing PNECs. These findings raise the possibility that increased neuropeptides would contribute to excess extravascular lung fluid and antagonizing their function may improve gas exchange.

2022-02-18 | GSE171907 | GEO

Transcriptional profiles of kidney upon HNF4A loss in mice by RNA-seq

Project description:Adult-onset knockout of HNF4A resulted in decreased expression levels of brush border genes. We find a robust shift in the transcriptome away from proximal tubule transcripts and towards distal tubule transcripts in the kidney upon HNF4A loss.

2021-03-15 | GSE157911 | GEO

Loss-of-function of MYO5B induces epithelial cell scattering in enterocytes

2013-03-14 | GSE45134 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data