Oncogenic targets Mmp7, S100a9, Nppb and Aldh1a3 from transcriptome profiling of FAP and Pirc adenomas are downregulated in response to tumor suppression by Clotam

Abstract

Intervention strategies in familial adenomatous polyposis (FAP) patients and other high-risk colorectal cancer (CRC) populations have highlighted a critical need for endoscopy combined with safe and effective preventive agents. We performed transcriptome profiling of colorectal adenomas from FAP patients and the polyposis in rat colon (Pirc) preclinical model, and prioritized molecular targets for prevention studies in vivo. At clinically relevant doses in the Pirc model, the drug Clotam (tolfenamic acid, TA) was highly effective at suppressing tumorigenesis both in the colon and in the small intestine, when administered alone or in combination with Sulindac. Cell proliferation in the colonic crypts was reduced significantly by TA, coincident with increased cleaved caspase-3 and decreased Survivin, β-catenin, cyclin D1 and matrix metalloproteinase 7. From the list of differentially expressed genes prioritized by transcriptome profiling, Mmp7, S100a9, Nppb and Aldh1a3 were defined as key oncogene candidates downregulated in colon tumors after TA treatment. Monthly colonoscopies revealed the rapid onset of tumor suppression by TA in the Pirc model, and the temporal changes in Mmp7, S100a9, Nppb and Aldh1a3, highlighting their value as potential early biomarkers for prevention in the clinical setting. We conclude that TA, an "old drug" repurposed from migraine, offers an exciting new therapeutic avenue in FAP and other high-risk CRC patient populations.

Heterocyclic Analogs of Sulforaphane Trigger DNA Damage and Impede DNA Repair in Colon Cancer Cells: Interplay of HATs and HDACs

“…Rats were administered a single oral gavage of 60 mg kg −1 of ITC, or vehicle. The study was terminated 6 h after dosing, rats were euthanized, and colon polyps were collected and stored prior to immunoblotting and immunohistochemistry (IHC), as reported …”

“…Formalin‐fixed paraffin‐embedded rat colon tumor was processed for IHC, as reported . Slides containing 5 μm sections were rehydrated and placed in an Autostainer (Dako).…”

Acetylation of CCAR2 Establishes a BET/BRD9 Acetyl Switch in Response to Combined Deacetylase and Bromodomain Inhibition

“…Library preparation via a NEBNext Ultra Directional RNA Library Prep Kit was followed by Illumina sequencing on a NextSeq 500/550 instrument (Illumina). Real-time reverse transcription quantitative PCR (RT-qPCR) was conducted according to a reported methodology (28).…”

“…Rats were then treated for 5 weeks with test agents, as follows: sulforaphane, 400 parts per million in AIN93 diet; JQ1, 12.5 mg/kg body weight via twice weekly intraperitoneal injection; sulforaphane þ JQ1, at the doses of the individual compounds, or vehicle. The study was terminated 2 months after polypectomy, and gastrointestinal lesions were enumerated prior to IB and RNA-seq, as reported previously (28). To our knowledge, this is the first report to examine secondary prevention in a murine model of FAP, following surgical intervention.…”

A miRNA signature for an environmental heterocyclic amine defined by a multi-organ carcinogenicity bioassay in the rat

“…Total RNA was extracted in Trizol reagent (Life Technologies), whereas, miRNA was extracted using the miRNeasy kit (Qiagen, Valencia, CA, USA). RNA quantity and purity were verified by Nanodrop ND-1000, from the absorbance at 260 and 280 nm (260/280 ratio >1.9), as reported (Simonich et al 2007; Jubert et al 2009; Ertem et al 2017). RNA (1 µg) from each rat tissue was reverse transcribed using the Superscript III RT kit (Life Technologies) in 10-µl reaction buffer, and diluted 10-fold before adding to a 10-µl reaction containing SYBR Green I Master mix (Roche Applied Science) and gene-specific primers.…”