Cargando…

SAT-033 Studying Cell-Context Specific Effects on the Function of the AT1R in iPSCs and Their Differentiated Derivatives

Cardiovascular homeostasis is tightly regulated by numerous neurohormonal mediators such as the renin-angiotensin system which plays an important role in the maintenance of blood pressure. Central to this system is the peptide hormone angiotensin II (Ang II) whose signals are transduced via the AT1...

Descripción completa

Detalles Bibliográficos
Autores principales: Bourque, Kyla, Devost, Dominic, Pétrin, Darlaine, Hébert, Terence
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Endocrine Society 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6552636/
http://dx.doi.org/10.1210/js.2019-SAT-033
_version_ 1783424638748983296
author Bourque, Kyla
Devost, Dominic
Pétrin, Darlaine
Hébert, Terence
author_facet Bourque, Kyla
Devost, Dominic
Pétrin, Darlaine
Hébert, Terence
author_sort Bourque, Kyla
collection PubMed
description Cardiovascular homeostasis is tightly regulated by numerous neurohormonal mediators such as the renin-angiotensin system which plays an important role in the maintenance of blood pressure. Central to this system is the peptide hormone angiotensin II (Ang II) whose signals are transduced via the AT1 receptor (AT1R), an important member of the superfamily of G protein-coupled receptors (GPCRs). Ang II binding results in receptor activation characterized by structural re-arrangements within the receptor structure and the subsequent activation of its cognate G protein partners. GPCRs are allosteric in nature and their biological activity is highly dependent on the cell context in which they are expressed(1). Changes in the cellular background such as the differential availability of G proteins and effector molecules including putative dimer partners can affect receptor conformation and function. As such, we are interested in understanding how AT1R conformation and signaling are modulated by the cell context in which it is expressed(1). In the past, studies that aimed at understanding signaling downstream of GPCRs mostly relied on heterologous expression systems such as HEK 293 cells because of their ease of culture. Such studies led to a ‘one size fits all’ notion that our findings could be reasonably extrapolated to guide drug discovery platforms relevant for human disease. However, it is clear that with the high rate of drug attrition, we need more physiologically relevant cellular models for studies of molecular signal transduction events to be translatable. With this in mind, we are generating iPSCs that stably express a panel of conformation-sensitive biosensors that reliably report on the conformational changes in the AT1R(1,2). Our biosensors use resonance energy transfer between a bioluminescent donor and a fluorescent acceptor (FlAsH) where agonist-mediated conformational changes can be recorded(1). Here, we will investigate how the conformation of the AT1R changes when expressed in AT1R-relevant cell types such as iPSC-derived cardiomyocytes and vascular smooth muscle cells. We will investigate how our conformational profiles differ in different iPSC-derived cell types in response to AT1R-specific agonists. Our goal is to gain a better mechanistic understanding of how cells are differentially wired leading to cell-specific conformational and signaling responses. We hope our results can guide rational drug design to better target the AT1R and other GPCRs. (1)Devost D., et al (2017). Journal of Biological Chemistry, jbc-M116. (2)Pei Y., et al (2015). Scientific reports, 5, 9205.
format Online
Article
Text
id pubmed-6552636
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Endocrine Society
record_format MEDLINE/PubMed
spelling pubmed-65526362019-06-13 SAT-033 Studying Cell-Context Specific Effects on the Function of the AT1R in iPSCs and Their Differentiated Derivatives Bourque, Kyla Devost, Dominic Pétrin, Darlaine Hébert, Terence J Endocr Soc Non-Steroid Hormone Signaling Cardiovascular homeostasis is tightly regulated by numerous neurohormonal mediators such as the renin-angiotensin system which plays an important role in the maintenance of blood pressure. Central to this system is the peptide hormone angiotensin II (Ang II) whose signals are transduced via the AT1 receptor (AT1R), an important member of the superfamily of G protein-coupled receptors (GPCRs). Ang II binding results in receptor activation characterized by structural re-arrangements within the receptor structure and the subsequent activation of its cognate G protein partners. GPCRs are allosteric in nature and their biological activity is highly dependent on the cell context in which they are expressed(1). Changes in the cellular background such as the differential availability of G proteins and effector molecules including putative dimer partners can affect receptor conformation and function. As such, we are interested in understanding how AT1R conformation and signaling are modulated by the cell context in which it is expressed(1). In the past, studies that aimed at understanding signaling downstream of GPCRs mostly relied on heterologous expression systems such as HEK 293 cells because of their ease of culture. Such studies led to a ‘one size fits all’ notion that our findings could be reasonably extrapolated to guide drug discovery platforms relevant for human disease. However, it is clear that with the high rate of drug attrition, we need more physiologically relevant cellular models for studies of molecular signal transduction events to be translatable. With this in mind, we are generating iPSCs that stably express a panel of conformation-sensitive biosensors that reliably report on the conformational changes in the AT1R(1,2). Our biosensors use resonance energy transfer between a bioluminescent donor and a fluorescent acceptor (FlAsH) where agonist-mediated conformational changes can be recorded(1). Here, we will investigate how the conformation of the AT1R changes when expressed in AT1R-relevant cell types such as iPSC-derived cardiomyocytes and vascular smooth muscle cells. We will investigate how our conformational profiles differ in different iPSC-derived cell types in response to AT1R-specific agonists. Our goal is to gain a better mechanistic understanding of how cells are differentially wired leading to cell-specific conformational and signaling responses. We hope our results can guide rational drug design to better target the AT1R and other GPCRs. (1)Devost D., et al (2017). Journal of Biological Chemistry, jbc-M116. (2)Pei Y., et al (2015). Scientific reports, 5, 9205. Endocrine Society 2019-04-30 /pmc/articles/PMC6552636/ http://dx.doi.org/10.1210/js.2019-SAT-033 Text en Copyright © 2019 Endocrine Society https://creativecommons.org/licenses/by-nc-nd/4.0/ This article has been published under the terms of the Creative Commons Attribution Non-Commercial, No-Derivatives License (CC BY-NC-ND; https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Non-Steroid Hormone Signaling
Bourque, Kyla
Devost, Dominic
Pétrin, Darlaine
Hébert, Terence
SAT-033 Studying Cell-Context Specific Effects on the Function of the AT1R in iPSCs and Their Differentiated Derivatives
title SAT-033 Studying Cell-Context Specific Effects on the Function of the AT1R in iPSCs and Their Differentiated Derivatives
title_full SAT-033 Studying Cell-Context Specific Effects on the Function of the AT1R in iPSCs and Their Differentiated Derivatives
title_fullStr SAT-033 Studying Cell-Context Specific Effects on the Function of the AT1R in iPSCs and Their Differentiated Derivatives
title_full_unstemmed SAT-033 Studying Cell-Context Specific Effects on the Function of the AT1R in iPSCs and Their Differentiated Derivatives
title_short SAT-033 Studying Cell-Context Specific Effects on the Function of the AT1R in iPSCs and Their Differentiated Derivatives
title_sort sat-033 studying cell-context specific effects on the function of the at1r in ipscs and their differentiated derivatives
topic Non-Steroid Hormone Signaling
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6552636/
http://dx.doi.org/10.1210/js.2019-SAT-033
work_keys_str_mv AT bourquekyla sat033studyingcellcontextspecificeffectsonthefunctionoftheat1rinipscsandtheirdifferentiatedderivatives
AT devostdominic sat033studyingcellcontextspecificeffectsonthefunctionoftheat1rinipscsandtheirdifferentiatedderivatives
AT petrindarlaine sat033studyingcellcontextspecificeffectsonthefunctionoftheat1rinipscsandtheirdifferentiatedderivatives
AT hebertterence sat033studyingcellcontextspecificeffectsonthefunctionoftheat1rinipscsandtheirdifferentiatedderivatives