The effects of neuroblastoma and chemotherapy on metabolism, fecal microbiome, volatile organic compounds, and gut barrier function in a murine model

Abstract

BACKGROUND: Following transplantation of human neuroblastoma (NB) cells into athymic mice, we investigated the effects of tumor growth and cyclophosphamide (CTX) treatment on systemic metabolism, gut inflammation and permeability, fecal microbiome and volatile organic compounds (VOCs). METHODS: NB cells (MHH-NB11) were implanted into athymic mice (n=20); 20 healthy mice served as controls (sham). CTX was given to 20 animals (10 NB and 10 sham) after 8 and 9 weeks. Metabolic changes were measured. Ileum samples were obtained for RT-PCR (claudins 2 and 4, occludin, tight junction protein 1) and apoptosis rate determination. Fecal microbiome and VOCs were analyzed. Values were compared to sham animals. RESULTS: NB caused reduction of adipose tissue, increases of IL-6 and TNF-α, and decreases of TGF-β1 and-β2. Serum FITCdextrane levels were increased in NB and improved under CTX. Claudin 4 expression was higher in NB versus NB + CTX and sham animals. NB caused increased apoptosis of epithelial cells. NB but also CTX led to a reduction in the abundance of Lactobacillus. NB led to alterations of the fecal VOC profile. CONCLUSIONS: NB caused a catabolic pro-inflammatory state, increased gut permeability, altered fecal VOCs and reductions of Lactobacillus. Further investigations are required to determine if modifications of the intestinal microbiome may reverse some of the observed effects.

The Effects of Prebiotic Supplementation with OMNi-LOGiC® FIBRE on Fecal Microbiome, Fecal Volatile Organic Compounds, and Gut Permeability in Murine Neuroblastoma-Induced Tumor-Associated Cachexia

“…5). Cells were cultured in RPMI medium (RPMI-1640 Medium, Sigma-Aldrich Handels GmbH, Vienna, Austria) with 2 mM glutamine supplemented with 10% fetal bovine serum (Sigma-Aldrich Handels GmbH, Vienna, Austria) and 1X minimum essential medium eagle (Sigma-Aldrich Handels GmbH, Vienna, Austria) nonessential amino acids as described previously [ 5 ]. After six passages, cell entity was verified by STR analysis with a Promega PowerPlex 16HS System (Promega GmbH, Braunschweig, Germany; Cat.…”

“…The gonadal, peri-renal, and inguinal white adipose tissue (WAT) were dissected, combined, and weighed as previously described [ 5 ]. The following organs were excised for further analysis: liver, spleen, kidney, triceps surae muscle, right tibia, and 3 cm ileum (beginning 1 cm orally of the ileocecal valve).…”

“…For VOC analysis, specimens were sent to our laboratory partner in Rostock by overnight express maintaining a temperature of 6-8 °C and examined within 48 h after harvesting. VOC analysis was performed in the headspace of stool samples as previously reported [ 5 , 26 , 27 ]. VOCs were pre-concentrated with a commercially available solid-phase microextraction (SPME) fiber (carboxen/polymethylsiloxane, Sulpeco, Bellefonte, PA, USA).…”

“…However, an interaction between host and tumor involving the secretion of pro-inflammatory cytokines from both cancer cells and the host immune system has been shown to be involved. These soluble mediators include tumor necrosis factor alpha (TNF-α), interferon gamma (INF-γ), and a variety of interleukins (IL-6, IL-8, IL-1β, and others) [ 4 , 5 , 6 ]. Additionally, the tumor itself may release catabolic substances such as proteolysis-inducing factor, anemia-inducing substance, activin, or myostatin, thereby further fueling the progression of TAC [ 4 , 7 ].…”

“…Although the tumor itself seems to play the major role in development and progression of TAC, the intestinal microbiome and its interaction with gut barrier, inflammation, and metabolism have also been implicated in the pathogenesis of TAC [ 5 , 8 , 9 , 10 , 11 , 12 ]. Among others, the intestinal microbiome serves as source for bacterial metabolites such as short-chain fatty acids and their derivatives, is involved in bile acid metabolism, and secretes microbe-associated molecular patterns (MAMPs) [ 13 , 14 , 15 ].…”