Summary of Study ST004082

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002563. The data can be accessed directly via it's Project DOI: 10.21228/M8BV8N This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST004082
Study Title	The secreted metabolite sensor CtBP2 links metabolism to healthy lifespan
Study Summary	While metabolic homeostasis is coordinated by metabolite-sensing molecules within a cell, their roles in horizontal regulation across cells remain underexplored. Here, we describe a system regulated by a metabolite sensor CtBP2. CtBP2 is secreted via exosomes in response to reductive metabolism, which is suppressed by oxidative stress. A series of pro-longevity agents induce CtBP2 secretion, and administration of exosomal CtBP2 extends lifespan of aged mice. Consistently, serum CtBP2 concentrations are higher in human subjects from families with a history of longevity and exhibit an age-dependent decline. Healthspan is also improved by exosomal CtBP2, as exemplified by protection against frailty in mice and negative associations between serum CtBP2 levels and cardiovascular diseases in humans. These health benefits could be attributed to the proper channeling of reducing equivalents into a chain of activation of CYB5R3 and AMPK. Our findings illustrate a previously uncharacterized metabolic system that provides insights into the regulation of human health and holds promise for future clinical applications.
Institute	University of Tsukuba
Last Name	Sekiya
First Name	Motohiro
Address	1-1-1 Tennodai, Tsukuba, Ibaraki, 3058575, Japan
Email	msekiya@md.tsukuba.ac.jp
Phone	+81298533053
Submit Date	2025-07-28
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	CE-MS
Release Date	2025-08-03
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002563
Project DOI:	doi: 10.21228/M8BV8N
Project Title:	IMR90 cells treated with exosomes with or without CtBP2
Project Summary:	We prepared two types of MEF (mouse embryonic fibroblast) cells: one is cells with wild-type CtBP2-HA knock-in and the other is cells with CtBP2 mutant-HA knock-in. The latter does not secrete CtBP2 in exosomes. We isolated exosomes from these cells and treated IMR90 fibroblasts (WT and Mut, respectively).
Institute:	University of Tsukuba
Last Name:	Sekiya
First Name:	Motohiro
Address:	1-1-1 Tennodai, Tsukuba, Ibaraki, 3058575, Japan
Email:	msekiya@md.tsukuba.ac.jp
Phone:	+81-29-853-3053

Subject:

Subject ID:	SU004228
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id	local_sample_id	Group	Sample source
SA473570	Mut-1	Mut	IMR90 fibroblasts
SA473571	Mut-2	Mut	IMR90 fibroblasts
SA473572	Mut-3	Mut	IMR90 fibroblasts
SA473573	WT-1	WT	IMR90 fibroblasts
SA473574	WT-2	WT	IMR90 fibroblasts
SA473575	WT-3	WT	IMR90 fibroblasts

Showing results 1 to 6 of 6

Showing results 1 to 6 of 6

Collection:

Collection ID:	CO004221
Collection Summary:	We identified an anti-aging effect of CtBP2 in exosomes. To investigate the mechanistic insights behind this, we decided to treat IMR90 fibroblasts with exosomes with or without CtBP2. Since we revealed that CtBP2 mutant lacking the metabolite sensing pocket cannot be secreted via exosomes, we prepared two types of MEF cells: one is cells with wild-type CtBP2-HA knock-in and the other is cells with the CtBP2 mutant-HA knock-in. We isolated exosomes from these cells and treated IMR90 fibroblasts for 1.5 days. Exosomes from MEF cells with wild-type CtBP2-HA knock-in contain CtBP2, and exosomes from those with mutant CtBP2-HA knock-in does not. WT and Mut denote metabolites extracted from IMR90 cells treated with exosomes with and without CtBP2, respectively.
Sample Type:	Fibroblast cells

Treatment:

Treatment ID:	TR004237
Treatment Summary:	IMR90 fibroblast cells were treated with exosomes with or without CtBP2, for 1.5 days in DMEM 10% exosome-free fetal bovine serum.

Sample Preparation:

Sampleprep ID:	SP004234
Sampleprep Summary:	Culture medium was aspirated from the dish, and the cells were washed twice with 5% mannitol solution (10 mL and 2 mL for the first and second wash, respectively). The cells were then treated with 800 µL of methanol and incubated at room temperature for 30 sec to suppress enzyme activity. Next, 550 µL of Milli-Q water containing internal standards (H3304-1002, Human Metabolome Technologies, Inc. (HMT), Tsuruoka, Yamagata, Japan) was added to the cell extract, followed by further incubation at room temperature for 30 sec. The cell extract was then centrifuged at 2,300 ×g, 4ºC for 5 min, after which 700 µL of the supernatant was centrifugally filtered through a Millipore 5-kDa cutoff filter (UltrafreeMC-PLHCC, HMT) at 9,100 ×g, 4ºC for 120 min to remove macromolecules. Subsequently, the filtrate was evaporated to dryness under vacuum and reconstituted in 50 µL of Milli-Q water for metabolome analysis at HMT.

Sampleprep ID:

SP004234

Sampleprep Summary:

Culture medium was aspirated from the dish, and the cells were washed twice with 5% mannitol solution (10 mL and 2 mL for the first and second wash, respectively). The cells were then treated with 800 µL of methanol and incubated at room temperature for 30 sec to suppress enzyme activity. Next, 550 µL of Milli-Q water containing internal standards (H3304-1002, Human Metabolome Technologies, Inc. (HMT), Tsuruoka, Yamagata, Japan) was added to the cell extract, followed by further incubation at room temperature for 30 sec. The cell extract was then centrifuged at 2,300 ×g, 4ºC for 5 min, after which 700 µL of the supernatant was centrifugally filtered through a Millipore 5-kDa cutoff filter (UltrafreeMC-PLHCC, HMT) at 9,100 ×g, 4ºC for 120 min to remove macromolecules. Subsequently, the filtrate was evaporated to dryness under vacuum and reconstituted in 50 µL of Milli-Q water for metabolome analysis at HMT.

Chromatography:

Chromatography ID:	CH005136
Instrument Name:	Agilent 7100 CE
Column Name:	Fused silica capillary (80 cm x 50 um)
Column Temperature:	N/A
Flow Gradient:	N/A
Flow Rate:	N/A
Solvent A:	Cation Buffer Solution (p/n:H3301-1001)
Solvent B:	N/A
Chromatography Type:	CE

Chromatography ID:	CH005137
Instrument Name:	Agilent 7100 CE
Column Name:	Fused silica capillary (80 cm x 50 um)
Column Temperature:	N/A
Flow Gradient:	N/A
Flow Rate:	N/A
Solvent A:	Anion Buffer Solution (p/n:I3302-1023)
Solvent B:	N/A
Chromatography Type:	CE

Analysis:

Analysis ID:	AN006759
Analysis Type:	MS
Chromatography ID:	CH005136
Num Factors:	2
Num Metabolites:	271
Units:	arbitrary unit

Analysis ID:	AN006760
Analysis Type:	MS
Chromatography ID:	CH005137
Num Factors:	2
Num Metabolites:	237
Units:	arbitrarry unit