Ultra-Deep Single-Cell Analyses Reveals Novel Mechanisms of Gut Enteroendocrine Cell Sensing Dysfunction in Human Obesity
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE154405 | GEO | 2021/11/30
REPOSITORIES: GEO
Similar Datasets
Project description:Croft2013 - GPCR-RGS interaction that compartmentalizes RGS activity
Through modelling studies, the classic quaternary complex (ligand-GPCR-G-RGS) has been extended to include an additional layer of regulation through GPCR-RGS interactions, which facilitate the compartmentalization of RGS activity into the plasma membrane and non-plasma compartments.
This model is described in the article:
A physiologically required G protein-coupled receptor (GPCR)-regulator of G protein signaling (RGS) interaction that compartmentalizes RGS activity.
Croft W, Hill C, McCann E, Bond M, Esparza-Franco M, Bennett J, Rand D, Davey J, Ladds G.
J Biol Chem. 2013 Sep 20;288(38):27327-42.
Abstract:
G protein-coupled receptors (GPCRs) can interact with regulator of G protein signaling (RGS) proteins. However, the effects of such interactions on signal transduction and their physiological relevance have been largely undetermined. Ligand-bound GPCRs initiate by promoting exchange of GDP for GTP on the Gα subunit of heterotrimeric G proteins. Signaling is terminated by hydrolysis of GTP to GDP through intrinsic GTPase activity of the Gα subunit, a reaction catalyzed by RGS proteins. Using yeast as a tool to study GPCR signaling in isolation, we define an interaction between the cognate GPCR (Mam2) and RGS (Rgs1), mapping the interaction domains. This reaction tethers Rgs1 at the plasma membrane and is essential for physiological signaling response. In vivo quantitative data inform the development of a kinetic model of the GTPase cycle, which extends previous attempts by including GPCR-RGS interactions. In vivo and in silico data confirm that GPCR-RGS interactions can impose an additional layer of regulation through mediating RGS subcellular localization to compartmentalize RGS activity within a cell, thus highlighting their importance as potential targets to modulate GPCR signaling pathways.
Author's comment on reproducing the plots:
To reproduce dose-response plots in the publication, the model is simulated with 12 different ligand concentrations (see parameter Ligand_conc).
For each ligand concentration, a single value corresponding to total amount of output must be obtained, by calculating the area under the curve of the trajectory of species z3, from time=0 to time=30.
These total output values are then used to build a dose-response plot (authors used GraphPad Prism).
Mutant strains are simulated with alternative parameter values or initial conditions specified in the Supplementary Material.
This model is hosted on BioModels Database
and identified
by: BIOMD0000000479
.
To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource
for published quantitative kinetic models
.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to the public
domain worldwide. Please refer to CC0 Public Domain
Dedication
for more information.
2024-09-02 | BIOMD0000000479 | BioModels
Project description:Bush2016 - Extended Carrousel model of
GPCR-RGS
This model is described in the article:
Yeast GPCR signaling
reflects the fraction of occupied receptors, not the
number.
Bush A, Vasen G, Constantinou A,
Dunayevich P, Patop IL, Blaustein M, Colman-Lerner A.
Mol. Syst. Biol. 2016 Dec; 12(12):
898
Abstract:
According to receptor theory, the effect of a ligand depends
on the amount of agonist-receptor complex. Therefore, changes
in receptor abundance should have quantitative effects.
However, the response to pheromone in Saccharomyces cerevisiae
is robust (unaltered) to increases or reductions in the
abundance of the G-protein-coupled receptor (GPCR), Ste2,
responding instead to the fraction of occupied receptor. We
found experimentally that this robustness originates during
G-protein activation. We developed a complete mathematical
model of this step, which suggested the ability to compute
fractional occupancy depends on the physical interaction
between the inhibitory regulator of G-protein signaling (RGS),
Sst2, and the receptor. Accordingly, replacing Sst2 by the
heterologous hsRGS4, incapable of interacting with the
receptor, abolished robustness. Conversely, forcing hsRGS4:Ste2
interaction restored robustness. Taken together with other
results of our work, we conclude that this GPCR pathway
computes fractional occupancy because ligand-bound GPCR-RGS
complexes stimulate signaling while unoccupied complexes
actively inhibit it. In eukaryotes, many RGSs bind to specific
GPCRs, suggesting these complexes with opposing activities also
detect fraction occupancy by a ratiometric measurement. Such
complexes operate as push-pull devices, which we have recently
described.
This model is hosted on
BioModels Database
and identified by:
BIOMD0000000638.
To cite BioModels Database, please use:
BioModels Database:
An enhanced, curated and annotated resource for published
quantitative kinetic models.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
2024-09-02 | BIOMD0000000638 | BioModels
Project description:Bush2016 - Simplified Carrousel model of
GPCR
This model is described in the article:
Yeast GPCR signaling
reflects the fraction of occupied receptors, not the
number.
Bush A, Vasen G, Constantinou A,
Dunayevich P, Patop IL, Blaustein M, Colman-Lerner A.
Mol. Syst. Biol. 2016 Dec; 12(12):
898
Abstract:
According to receptor theory, the effect of a ligand depends
on the amount of agonist-receptor complex. Therefore, changes
in receptor abundance should have quantitative effects.
However, the response to pheromone in Saccharomyces cerevisiae
is robust (unaltered) to increases or reductions in the
abundance of the G-protein-coupled receptor (GPCR), Ste2,
responding instead to the fraction of occupied receptor. We
found experimentally that this robustness originates during
G-protein activation. We developed a complete mathematical
model of this step, which suggested the ability to compute
fractional occupancy depends on the physical interaction
between the inhibitory regulator of G-protein signaling (RGS),
Sst2, and the receptor. Accordingly, replacing Sst2 by the
heterologous hsRGS4, incapable of interacting with the
receptor, abolished robustness. Conversely, forcing hsRGS4:Ste2
interaction restored robustness. Taken together with other
results of our work, we conclude that this GPCR pathway
computes fractional occupancy because ligand-bound GPCR-RGS
complexes stimulate signaling while unoccupied complexes
actively inhibit it. In eukaryotes, many RGSs bind to specific
GPCRs, suggesting these complexes with opposing activities also
detect fraction occupancy by a ratiometric measurement. Such
complexes operate as push-pull devices, which we have recently
described.
This model is hosted on
BioModels Database
and identified by:
BIOMD0000000637.
To cite BioModels Database, please use:
BioModels Database:
An enhanced, curated and annotated resource for published
quantitative kinetic models.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
2024-09-02 | BIOMD0000000637 | BioModels
Project description:Enteroendocrine cells (EECs) are hormone-producing cells residing in the epithelium of stomach, small intestine (SI) and colon. EECs regulate various aspects of metabolic activity including insulin levels, satiety, gastrointestinal secretion and motility. The generation of different EEC lineages of the SI is incompletely understood. We now report a TFome-wide CRISPR knockout screen in adult human intestinal organoids to identify dominant transcription factors controlling EEC differentiation. ZNF800 is discovered as a master repressor for endocrine lineage commitment, which particularly restricts enterochromaffin cell differentiation by directly controlling an endocrine TF network centered on PAX4. Thus, organoid models allow unbiased functional SCRISPR screens for genes that program cell differentiation.
2023-10-27 | GSE229584 | GEO
Project description:Enteroendocrine cells (EECs) are hormone-producing cells residing in the epithelium of stomach, small intestine (SI) and colon. EECs regulate various aspects of metabolic activity including insulin levels, satiety, gastrointestinal secretion and motility. The generation of different EEC lineages of the SI is incompletely understood. We now report a TFome-wide CRISPR knockout screen in adult human intestinal organoids to identify dominant transcription factors controlling EEC differentiation. ZNF800 is discovered as a master repressor for endocrine lineage commitment, which particularly restricts enterochromaffin cell differentiation by directly controlling an endocrine TF network centered on PAX4. Thus, organoid models allow unbiased functional SCRISPR screens for genes that program cell differentiation.
2023-10-27 | GSE229583 | GEO
Project description:Enteroendocrine cells (EECs) are hormone-producing cells residing in the epithelium of stomach, small intestine (SI) and colon. EECs regulate various aspects of metabolic activity including insulin levels, satiety, gastrointestinal secretion and motility. The generation of different EEC lineages of the SI is incompletely understood. We now report a TFome-wide CRISPR knockout screen in adult human intestinal organoids to identify dominant transcription factors controlling EEC differentiation. ZNF800 is discovered as a master repressor for endocrine lineage commitment, which particularly restricts enterochromaffin cell differentiation by directly controlling an endocrine TF network centered on PAX4. Thus, organoid models allow unbiased functional SCRISPR screens for genes that program cell differentiation.
2023-10-27 | GSE229585 | GEO
Project description:Hypothalamic neurons expressing Agouti-related peptide (AgRP) are critical for initiating food intake, but druggable biochemical pathways that control this response remain elusive. Thus, genetic ablation of insulin or leptin signaling in AgRP neurons is predicted to reduce satiety but fails to do so. FoxO1 is a shared mediator of both pathways, and its inhibition is required to induce satiety. Accordingly, FoxO1 ablation in AgRP neurons of mice results in reduced food intake, leanness, improved glucose homeostasis, and increased sensitivity to insulin and leptin. Expression profiling of flow-sorted FoxO1-deficient AgRP neurons identifies G-protein-coupled receptor Gpr17 as a FoxO1 target whose expression is regulated by nutritional status. Intracerebroventricular injection of Gpr17 agonists induces food intake, whereas Gpr17 antagonist cangrelor curtails it. These effects are absent in Agrp-Foxo1 knockouts, suggesting that pharmacological modulation of this pathway has therapeutic potential to treat obesity. We used microarrays to detail the change of gene expression in AgRP neurons after knocking out FoxO1. AgRP neurons from control and KO mice were collected by FACS. Gene expression was analyzed by microarray.
2013-04-09 | E-GEOD-45858 | biostudies-arrayexpress
Project description:Adjacent stroma, including subepithelial fibroblasts, are believed to coordinate the differentiation process of epithelial cells, but the mechanisms are not well understood. Glial cell line-derived neurotrophic factor (GDNF) is expressed in the intestinal Pdgfra high subepithelial myofibroblasts (SEMFs), while the GDNF receptor RET is expressed in a subset of enteroendocrine cells (EECs), indicating regulatory crosstalk. In this experiment, single RET+ intestinal cells were sorted the gene expression profile was compared to non-RET cells. This provided knowledge about RET+ cell types. RET+ cells were upregulated with most of the EEC markers, indicating that RET+ cells are expressed in EECs. We also observed the differences in the gene expression profile of stromal-derived GDNF ligand on intestinal organoids. Organoids were derived from intestinal crypts of WT mice C57BL/6J and treated with 500ng/ml of Gdnf and Gfra1. GDNF-treated organoids induced the expression of EEC genes such as Pyy, Tac1, Tph1, and Cck, indicating enhanced differentiation of EC cell and L-I-N cell lineages. This highlights a stroma-epithelium crosstalk pathway regulating the differentiation of intestinal EEC subtypes.
2024-10-17 | GSE259326 | GEO
Project description:Regulators of G protein signaling (RGS proteins) constrain G protein-coupled receptor (GPCR)-mediated and other responses throughout the body primarily, but not exclusively, through their GTPase activating (GAP) activity. Asthma is a highly prevalent condition characterized by airway hyper-responsiveness (AHR) to environmental stimuli resulting in part from amplified GPCR-mediated airway smooth muscle (ASM) contractility. Rgs2 or Rgs5 gene deletion in mice enhances AHR and ASM contraction whereas RGS4 knockout (KO) mice unexpectedly have decreased AHR due to increased production of the bronchodilator prostaglandin E2 (PGE2) by lung epithelial cells. Here we found that knockin mice harboring Rgs4 alleles encoding a point mutation (N128A) that sharply curtails RGS4 GAP activity had increased AHR, reduced airway PGE2 levels, and augmented GPCR-induced bronchial contraction compared to either RGS4 KO mice or WT controls. RGS4 interacted with the p85a subunit of PI3 kinase (PI3K) and inhibited PI3K-dependent PGE2 secretion elicited by TGFb in airway epithelial cells. Together these findings suggest that RGS4 affects asthma severity in part by regulating the airway inflammatory milieu in a G protein-independent manner.
2024-05-08 | GSE242546 | GEO
Project description:Hypothalamic neurons expressing Agouti-related peptide (AgRP) are critical for initiating food intake, but druggable biochemical pathways that control this response remain elusive. Thus, genetic ablation of insulin or leptin signaling in AgRP neurons is predicted to reduce satiety but fails to do so. FoxO1 is a shared mediator of both pathways, and its inhibition is required to induce satiety. Accordingly, FoxO1 ablation in AgRP neurons of mice results in reduced food intake, leanness, improved glucose homeostasis, and increased sensitivity to insulin and leptin. Expression profiling of flow-sorted FoxO1-deficient AgRP neurons identifies G-protein-coupled receptor Gpr17 as a FoxO1 target whose expression is regulated by nutritional status. Intracerebroventricular injection of Gpr17 agonists induces food intake, whereas Gpr17 antagonist cangrelor curtails it. These effects are absent in Agrp-Foxo1 knockouts, suggesting that pharmacological modulation of this pathway has therapeutic potential to treat obesity. We used microarrays to detail the change of gene expression in AgRP neurons after knocking out FoxO1.
2013-04-09 | GSE45858 | GEO