The purpose of this research is to determine how exercise affects fat (adipose) tissue and how changes to adipose tissue that occur during and after exercise might improve health in aging and obesity.
Adipose tissue has important endocrine functions that influence metabolic health. Early evidence shows that adipose adapts to physiological stress, including exercise. The objective here is to determine how exercise-induced alterations in adipose tissue cellular composition and endocrine signaling may contribute to the beneficial adaptations to exercise in aging and obesity. Immune cell populations and inflammatory signatures will be assessed in subcutaneous abdominal adipose tissue biopsies collected from obese and normal weight young and older adults before, immediately after, and 3 hours after a 30-min bout of cycling exercise at 70% of maximal oxygen consumption (VO2max). Mass spectrometry, Olink targeted inflammation assays, and RNA sequencing will be used for full proteomic and transcriptomic characterization of the adipose tissue secretome (the proteins and molecules secreted from the adipose tissue) and the cargo of extracellular vesicles isolated from plasma and media collected from cultured human adipose tissue explants generated from each time point. Overall, the primary hypothesis of the proposed work is that a single bout of exercise triggers transient changes in adipose tissue paracrine/endocrine signals and immune cellular composition. The investigators propose that these responses contribute to the beneficial effects of exercise locally and in distal tissues and that the cumulative effects of acute changes in adipose tissue likely contribute to the positive alterations in adipose tissue associated with exercise training.
Study Type
INTERVENTIONAL
Allocation
NON_RANDOMIZED
Purpose
BASIC_SCIENCE
Masking
NONE
Enrollment
56
Fat sample collected from abdomen before exercise, immediately after exercise, and 3 hours after exercise
30 minutes of exercise on a cycle ergometer exercise machine
Mayo Clinic
Rochester, Minnesota, United States
Adipose lymphocyte populations
The numbers of lymphocytes in the adipose tissue will be assessed by flow cytometry.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
IL6 secreted from adipose tissue measured using the Olink Target Inflammation Panel (includes 96 inflammation-associated proteins)
The concentration of 96 IL6 will be measured using the targeted inflammation Olink proximity extension assay panel in media obtained from cultured adipose tissue explants. Adipose tissue samples will be cultured for 3 hours, and the culture media will be removed for the assessment of secreted inflammatory proteins. T
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue macrophage populations
Numbers of macrophages in the adipose tissue will be assessed by immunohistochemistry.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of interleukin (IL)-6 measured by polymerase chain reaction (PCR).
Gene expression of the inflammatory cytokine IL-6 will be measured in adipose tissue samples using real time (RT)-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of interleukin (IL)-8 measured by PCR.
Gene expression of the inflammatory cytokine IL-8 will be assessed in adipose tissue samples using real-time (RT)-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of cluster of differentiation 68 (CD68) measured by PCR.
Gene expression of the macrophage marker CD68 be assessed in adipose tissue samples using real time (RT)-PCR.
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Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of cluster of differentiation 163 (CD163) measured by PCR.
Gene expression of the anti-inflammatory macrophage marker CD163 will be assessed in adipose tissue samples using RT-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of cluster of differentiation 206 (CD206) measured by PCR.
Gene expression of the anti-inflammatory macrophage marker CD206 will be assessed in adipose tissue samples using RT-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of adhesion G-protein coupled receptor E (ADGRE) measured by PCR.
Gene expression of the macrophage marker ADGRE will be assessed in adipose tissue samples using RT-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of inducible nitric oxide synthase (INOS) measured by PCR.
Gene expression of the pro-inflammatory macrophage marker INOS will be assessed in adipose tissue samples using RT-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of monocyte chemoattractant protein-1 (MCP1) measured by PCR.
Gene expression of the inflammatory cytokine MCP1 will be assessed in adipose tissue samples using RT-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of tumor necrosis factor alpha (TNFa) measured by PCR.
Gene expression of the inflammatory cytokine TNFa will be assessed in adipose tissue samples using RT-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of leptin (LEPD) measured by PCR.
Gene expression of the pro-inflammatory adipokine LEPD will be assessed in adipose tissue samples using RT-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipose tissue gene expression of adiponectin (ADIPOQ) measured by PCR.
Gene expression of the anti-inflammatory adipokine ADIPOQ will be assessed in adipose tissue samples using RT-PCR.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Inflammatory proteins found in adipose tissue measured using the Olink Target Inflammation Panel (includes 96 inflammation-associated proteins)
The concentrations of 96 different inflammatory proteins (including various cytokines and adipokines) will be measured in adipose tissue lysates using the targeted inflammation Olink proximity extension assay panel. This assay measures all 96 proteins in a single assay and all are reported as normalized protein expression values.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Protein cargo of adipose extracellular vesicles measured using mass spectrometry
Adipose tissue samples will be cultured for 3 hours, and the culture media will be removed. From this media, extracellular vesicles will be isolated. In these extracellular vesicles, mass spectrometry will be performed to examine the entire proteome (all proteins) contained within these extracellular vesicles. Using the large number of proteins identified, the investigators will examine changes in categories of protein types (e.g., inflammatory proteins, metabolic proteins, transcriptional proteins).
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Adipocyte cell size
Adipocyte cell size will be determined using methylene blue staining and microscopy.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Plasma IL6 concentrations measured using the Olink Target Inflammation Panel
The concentration of IL6 will be measured in plasma samples using the targeted inflammation Olink proximity extension assay panel. This assay measures 96 proteins in a single assay and all are reported as normalized protein expression values.
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise
Protein cargo of plasma extracellular vesicles measured using mass spectrometry extracellular vesicles
Extracellular vesicles will be isolated from plasma samples. In these extracellular vesicles, mass spectrometry will be performed to examine the entire proteome (all proteins) contained within these extracellular vesicles. Using the large number of proteins identified, the investigators will examine changes in categories of protein types (e.g., inflammatory proteins, metabolic proteins, transcriptional proteins).
Time frame: Pre-exercise, immediately post-exercise, and 3 hours after exercise