Project description:Primary objectives: The primary objective is to investigate circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Primary endpoints: circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Project description:We employed whole genome gene expression analysis to characterize the intestinal exposure to 5 closely related food contaminants belonging to the type B trichocene mycotoxins groups The few data available on fusarenon-X (FX) do no support derivation of health-based guidance values for this mycotoxin, although preliminary results suggest higher toxicity compared to other regulated trichothecenes. Using histo-morphological analysis and whole-transcriptome profiling, the present study was designed to get a global view of intestinal alterations induced by FX. The well-described trichothecene deoxynivalenol (DON) served as a benchmark. FX exposure induced more severe intestinal histological alterations compared to DON. Intestinal inflammation was the hallmark of the molecular toxicity of DON, but also of FX. The dose-response analysis for FX revealed that benchmark doses for up-regulation of key-inflammatory genes expression were 4 to 45-fold higher than the previously reported ones for DON. Transcriptome analysis revealed that both mycotoxins down-regulated PPAR and LXR-RXR signaling pathways controlling lipid metabolism. Interestingly, several pathways including VDR/RXR activation, ephrin receptor signaling, and GNRH signaling were specific to FX and thus, discriminate the intestinal transcriptomic fingerprint of the two mycotoxins. Altogether, these results demonstrate that FX induces a more potent intestinal inflammation than DON. This study also reveals specific FX-targeted pathways, indicating that the toxicity of DON cannot serve as a benchmark for FX, and that toxicity evaluation of each trichothecene should be conducted separately.
Project description:Genome-wide DNA methylation profiling of white blood cells was conducted using the Illumina Infinium Human Methylation 850K Genechip. According to the internal exposure level of deoxynivalenol, 32 participants were divided into two groups (high exposure DON group and low exposure DON group). Samples included 16 peripheral blood samples from children with high DON exposure and 16 peripheral blood samples from children with low DON exposure.
Project description:Deoxynivalenol (DON) frequently detected in a wide range of foods and feeds, inducing cytotoxicity to animals and humans. N6-methyladenosine (m6A) is an important epitranscriptomic marker with high abundance in eukaryotic mammals mRNAs. However, the role of the m6A methylomes in DON damage is still poorly understood. Here, we investigated the m6A transcriptome-wide profile in intestinal porcine epithelial cells (IPEC-J2) with and without 1000 ng/mL DON treatment via m6A sequencing and RNA sequencing. In total, 5406 new m6A peaks appeared with the disappearance of 2615 peaks in DON-induced IPEC-J2. The unique m6A-modified genes in DON-induced IPEC-J2 were associated with TNF signaling pathway. We identified 733 differentially expressed mRNA transcripts with hyper-methylated or hypo-methylated m6A peaks between DON-induced IPEC-J2 and normal IPEC-J2. Protein interaction network analysis and qPCR validation suggested that CSF2 probably acts as a promising new target for combating DON damage in IPEC-J2. Our first report of m6A transcriptome-wide map of IPEC-J2 cells presented here provides a starting roadmap for uncovering m6A functions that may affect DON infection.
Project description:Fusarium graminearum (F. graminearum) and its derivative mycotoxin deoxynivalenol (DON) are highly concerned with food security, safety and sustainability worldwide. The molecular mechanism regulates DON biosynthesis in F. graminearum remains enigmatic, despite several transcription factors (TFs) have been revealed containing regulatory functions. Here, we first characterized a zinc finger-contained TF, FgSfp1. Interestingly, contrast to the previous knowledge, all TRI-cluster genes were abnormally upregulated in ∆FgSfp1 while Tri proteins abundance rationally decreased, resulting in a 95.4% reduction of DON yield simultaneously. Further evidence determined FgSfp1 acted as a nutrient-sensing factor coordinates genetic expression efficiency through interference of ribosomal biogenesis process. Specifically, FgSfp1-depletion leads to ribosome biogenesis assembly factor (RiBi) expression attenuation along with DON precursor acetyl-CoA synthase reduction since FgSfp1 actively interacts with RNA 2’-O-methylation enzyme FgNop1 revealed by Bi-FC and subsequently influences mRNA translation capacity. In conclusion, we elucidated that the FgSfp1 orchestrates DON biosynthesis via participating RNA posttranscriptional modification for ribosomal RNA maturation, offering new insights into the DON biosynthesis regulation. Ultimately, this novel TF might be a key regulator for DON contamination control in the whole food chain.
Project description:To gain mechanistic insight into how Epigenetic factors reprogramming metabolism in response to DON treatment. we performed CUT&Tag sequencing in murine PDAC cells. sgPaxip1 cells were treated in Cont group or DON group , and harvested after 72 hours. CUT&Tag assay was performed following the manual of hyperactive pG-Tn5/pA-Tn5 transposase for CUT&Tag kit (TD901, Vazyme). DNA library were prepared according to manufacturer’s instructions of Trueprep index kit v2 (TD202, Vazyme).