In vivo transcriptomic profiling using cell encapsulation identifieseffector pathways of systemic aging
Abstract
Sustained exposure to a young systemic environment rejuvenates aged organisms and promotes cellular function. However, due to the intrinsic complexity of tissues it remains challenging to pinpoint niche-independent effects of circulating factors on specific cell populations. Here we describe a method for the encapsulation of human and mouse skeletal muscle progenitors in diffusible polyethersulfone hollow fiber capsules that can be used to profile systemic aging in vivo independent of heterogeneous short-range tissue interactions. We observed that circulating long-range signaling factors in the old systemic environment lead to an activation of Myc and E2F transcription factors, induce senescence and suppress myogenic differentiation. Importantly, in vitro profiling using young and old serum in 2D culture does not capture all pathways deregulated in encapsulated cells in aged mice. Thus, in vivo transcriptomic profiling using cell encapsulation allows for the characterization of effector pathways of systemic aging with unparalleled accuracy.
Data availability
The data discussed in this publication have been deposited in NCBI's Gene Expression Omnibus (GEO). GEO Series accession numbers are GSE111401, GSE81096 and GSE193665.
-
In Vivo Transcriptomic Profiling using Cell Encapsulation Identifies Effector Pathways of Systemic AgingNCBI Gene Expression Omnibus, GSE111401.
-
In Vivo Transcriptomic Profiling using Cell Encapsulation Identifies Effector Pathways of Systemic AgingNCBI Gene Expression Omnibus, GSE193665.
Article and author information
Author details
Funding
Canadian Institutes of Health Research (PJT-162442)
- C Florian Bentzinger
Association Française contre les Myopathies (AFM)
- Pura Muñoz-Cánoves
MWRF (MWRF)
- Pura Muñoz-Cánoves
Maria de Maeztu Unit of Excellence award to UPF (MDM-2014-0370)
- Pura Muñoz-Cánoves
Severo Ochoa Center of Excellence award to the CNIC (SEV-2015-0505)
- Pura Muñoz-Cánoves
Severo Ochoa FPI predoctoral fellowship (SEV-2015-0505-17-1))
- Xiaotong Hong
National Science and Research Council of Canada (RGPIN-2017-05490)
- C Florian Bentzinger
Fonds de Recherche du Québec - Santé (Dossiers 296357,34813,and 36789)
- C Florian Bentzinger
Centre de Recherche Médicale de l'Université de Sherbrooke (CRMUS Chair)
- C Florian Bentzinger
European Research Council (ERC-2016-AdG-741966)
- Pura Muñoz-Cánoves
La Caixa Foundation (La Caixa-HEALTH-HR17-00040)
- Pura Muñoz-Cánoves
Muscular Dystrophy Association (MDA)
- Pura Muñoz-Cánoves
H2020 (UPGRADE-H2020-825825)
- Pura Muñoz-Cánoves
Programa Estatal de Investigacion (RTI2018-096068-B-I00)
- Pura Muñoz-Cánoves
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was performed in accordance with the Swiss regulation on animal experimentation and the European Community Council directive (86/609/EEC) for the care and use of laboratory animals. Experiments were approved by the Vaud cantonal authorities under license VD3085, and by the Animal Care and Ethics Committee of the Spanish National Cardiovascular Research Center (CNIC) and regional authorities.
Copyright
© 2022, Mashinchian et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 2,252
- views
-
- 339
- downloads
-
- 1
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Further reading
-
- Cell Biology
- Plant Biology
Autophagy-related gene 6 (ATG6) plays a crucial role in plant immunity. Nonexpressor of pathogenesis-related genes 1 (NPR1) acts as a signaling hub of plant immunity. However, the relationship between ATG6 and NPR1 is unclear. Here, we find that ATG6 directly interacts with NPR1. ATG6 overexpression significantly increased nuclear accumulation of NPR1. Furthermore, we demonstrate that ATG6 increases NPR1 protein levels and improves its stability. Interestingly, ATG6 promotes the formation of SINCs (SA-induced NPR1 condensates)-like condensates. Additionally, ATG6 and NPR1 synergistically promote the expression of pathogenesis-related genes. Further results showed that silencing ATG6 in NPR1-GFP exacerbates Pst DC3000/avrRps4 infection, while double overexpression of ATG6 and NPR1 synergistically inhibits Pst DC3000/avrRps4 infection. In summary, our findings unveil an interplay of NPR1 with ATG6 and elucidate important molecular mechanisms for enhancing plant immunity.
-
- Cell Biology
Prostaglandin E2 (PGE2) is an endogenous inhibitor of glucose-stimulated insulin secretion (GSIS) and plays an important role in pancreatic β-cell dysfunction in type 2 diabetes mellitus (T2DM). This study aimed to explore the underlying mechanism by which PGE2 inhibits GSIS. Our results showed that PGE2 inhibited Kv2.2 channels via increasing PKA activity in HEK293T cells overexpressed with Kv2.2 channels. Point mutation analysis demonstrated that S448 residue was responsible for the PKA-dependent modulation of Kv2.2. Furthermore, the inhibitory effect of PGE2 on Kv2.2 was blocked by EP2/4 receptor antagonists, while mimicked by EP2/4 receptor agonists. The immune fluorescence results showed that EP1–4 receptors are expressed in both mouse and human β-cells. In INS-1(832/13) β-cells, PGE2 inhibited voltage-gated potassium currents and electrical activity through EP2/4 receptors and Kv2.2 channels. Knockdown of Kcnb2 reduced the action potential firing frequency and alleviated the inhibition of PGE2 on GSIS in INS-1(832/13) β-cells. PGE2 impaired glucose tolerance in wild-type mice but did not alter glucose tolerance in Kcnb2 knockout mice. Knockout of Kcnb2 reduced electrical activity, GSIS and abrogated the inhibition of PGE2 on GSIS in mouse islets. In conclusion, we have demonstrated that PGE2 inhibits GSIS in pancreatic β-cells through the EP2/4-Kv2.2 signaling pathway. The findings highlight the significant role of Kv2.2 channels in the regulation of β-cell repetitive firing and insulin secretion, and contribute to the understanding of the molecular basis of β-cell dysfunction in diabetes.