Metabolomic profiling reveals muscle metabolic changes following iliac arteriovenous fistula creation in mice

Abstract

End-stage kidney disease (ESKD), the most advanced stage of chronic kidney disease (CKD), requires renal replacement therapy or kidney transplant to sustain life. To accomplish durable dialysis access, the creation of an arteriovenous fistula (AVF) has emerged as a preferred approach. Unfortunately, a significant proportion of patients that receive an AVF experience some form of hand dysfunction, however the mechanisms underlying these side effects are not understood. In this study, we utilized nuclear magnetic resonance (NMR) spectroscopy to investigate the muscle metabolome following iliac AVF placement in mice with CKD. To induce CKD, C57BL6J mice we fed an adenine-supplemented diet for three weeks and then randomized to receive AVF or Sham surgery. Two-weeks following surgery, the quadriceps muscles were rapidly dissected and snap frozen for metabolite extraction and subsequent NMR analysis. Principal component analysis demonstrated clear separation between groups, confirming a unique metabolome in mice that received an AVF. AVF creation resulted in reduced levels creatine, ATP, ADP, as well as increased levels of IMP and several tricarboxylic acid cycle metabolites suggesting profound energetic stress. Pearson correlation and multiple linear regression analyses identified several metabolites that were strongly linked to measures of limb function (grip strength, gait speed, mitochondrial respiration). In summary, AVF creation generates a unique metabolome profile in the distal skeletal muscle indicative of an energetic crisis and myosteatosis.

Temporal serum metabolomic and lipidomic analyses distinguish patients with access-related hand disability following arteriovenous fistula creation

“…It is well known that the kidney plays a crucial role in the uptake of phenylalanine and its conversion to tyrosine 29 , 30 . Interestingly, our recent analysis of the muscle metabolome in a murine AVF model also identified that high levels of muscle phenylalanine were associated with weakness (hindlimb grip strength) and motor disability (walking speed) 16 , suggesting that serum phenylalanine may be a biomarker for these phenotypic characteristics of ARHD.…”

“…A 600 MHz Bruker Avance III system (Bruker BioSpin Corporation, Billerica, MA, USA) with a 5-mm TCl CryoProbe was used to acquire all 1-dimensional (1D) and 2-dimensional (2D) NMR spectra. 1D 1 H spectra were acquired with the 1D nuclear Overhauser effect spectroscopy (NOESY) pulse sequence (noesypr1d) 15 , utilizing identical parameters described previously 13 , 14 , 16 . Two dimensional spectra including heteronuclear single quantum coherence (HSQC), heteronuclear multiple bond coherence (HMBC), correlated spectroscopy (COSY), and total correlated spectroscopy (TOCSY) were acquired using the standard Bruker library, as well as previous work 17 to confirm metabolite identity.…”

Temporal serum metabolomic and lipidomic analyses distinguish patients with access-related hand disability following arteriovenous fistula creation

“…It is well known that the kidney plays a crucial role in the uptake of phenylalanine and its conversion to tyrosine 28,29 . Interestingly, our recent analysis of the muscle metabolome in a murine AVF model also identi ed that high levels of muscle phenylalanine were associated with weakness (hindlimb grip strength) and motor disability (walking speed) 30 , suggesting that serum phenylalanine may be a biomarker for these phenotypic characteristics of ARHD.…”

Mitochondrial targeted catalase improves muscle strength following arteriovenous fistula creation in mice with chronic kidney disease

“…One week following arrival, all mice were switched to a 20% casein-based chow diet. After 1 week on casein diet, all mice were fed a 0.2% adenine-supplemented casein chow to induce chronic kidney disease prior to the AVF surgery and remained on the adenine diet throughout the study as previously described 18 , 23 , 24 , 32 – 35 . All animals were provided both food and water ad libitum for the duration of the experiment.…”

“…To begin to explore whether skeletal muscle pathology contributes to ARHD following AVF creation, our group developed a novel mouse model in which an AVF can be created in the iliac vascular bundle allowing examination of limb pathophysiology 18 , 19 which is not possible using alternative pre-clinical models using aortic or carotid vessels 20 – 22 . In this murine iliac AVF model, mice have been shown to exhibit hindlimb weakness, gait abnormalities, and muscle mitochondrial abnormalities 18 , 23 , 24 . Oxidative stress and mitochondrial dysfunction have been frequently reported in the skeletal muscle of rodents and human patients with CKD 10 – 12 , 16 , 25 – 28 .…”