Search

Colorectal cancer causes over 53,000 deaths annually in the United States. Its rising incidences worldwide and particularly in young adults is a major concern. Here, we evaluated the efficacy of omeprazole that is clinically approved for treating acid reflux, to enable its repurposing for colorectal cancer prevention. In the azoxymethane-induced rat colorectal cancer model, dietary omeprazole (250 and 500 ppm) was administered at early adenoma stage (8 weeks after azoxymethane) to assess the progression of early lesions to adenocarcinoma. Administration of omeprazole at 250 or 500 ppm doses led to suppression of total colon adenocarcinoma incidence by 15.7% and 32% (P < 0.01), respectively. Importantly, invasive carcinoma incidence was reduced by 59% (P < 0.0005) and 90% (P < 0.0001) in omeprazole-administered rats in a dose-dependent manner. There was also a strong and dose-dependent inhibition in the adenocarcinoma multiplicity in rats exposed to omeprazole. Administration of 250 and 500 ppm omeprazole inhibited total colon adenocarcinoma multiplicity by approximately 49% and approximately 65% (P < 0.0001), respectively. While noninvasive adenocarcinomas multiplicity was suppressed by approximately 34% to approximately 48% (P < 0.02), the invasive carcinomas multiplicity was reduced by approximately 74% to approximately 94% (P < 0.0001) in omeprazole-exposed rats in comparison with the untreated rats. Biomarker analysis results showed a decrease in cell proliferation and anti-apoptotic/pro-survival proteins with an increase in apoptosis. Transcriptome analysis of treated tumors revealed a significant increase in adenocarcinoma inhibitory genes (Olmf4; Spink4) expression and downregulation of progression promoting genes (SerpinA1, MMP21, IL6). In summary, omeprazole showed significant protection against the progression of adenoma to adenocarcinoma.Prevention Relevance:Preventing colon cancer is urgently needed because of its high incidence and mortality rates worldwide. Toward this end, preventive efficacy of omeprazole, a common medication, was evaluated in animal model of colorectal cancer and was found to suppress colonic adenoma progression to carcinoma. These findings warrant its further evaluation in humans.

Daily dosing of either NSAIDs or EGFR inhibitors has been shown to prevent bladder cancer development in a N-butyl-(4-hydroxybutyl)nitrosamine (OH-BBN)-induced rat model. However, these inhibitors cause gastrointestinal ulceration and acneiform rash, respectively, limiting their continuous use in a clinical prevention setting. We studied chemopreventive efficacy of pulsatile dosing of EGFR inhibitor erlotinib (42 mg/kg BW, once/week) combined with intermittent or continuous low doses of the NSAID naproxen (30 mg/kg BW/day, 3 weeks on/off or 128 ppm daily in diet) in the OH-BBN induced rat bladder cancer model. The interventions were started either at 1 or 4 weeks (early intervention) or 3 months (delayed intervention) after the last OH-BBN treatment, by which time the rats had developed microscopic bladder lesions. All combination regimens tested as early versus late intervention led to the reduction of the average bladder tumor weights (54%–82%; P < 0.01 to P < 0.0001), a decrease in tumor multiplicity (65%–85%; P < 0.01 to P < 0.0001), and a decrease in the number of rats with large palpable tumors (>200 mg; 83%–90%; P < 0.01 to P < 0.0001). Levels of signal transduction markers, Ki-67, cyclin D1, IL1β, pSTAT3, and pERK, were significantly (P < 0.05 to P < 0.001) reduced in the treated tumors, demonstrating their potential utility as predictive markers for efficacy. These findings demonstrate that significant chemopreventive efficacy could be achieved with alternative intervention regimens designed to reduce the toxicity of agents, and that starting erlotinib and/or naproxen treatments at the time microscopic tumors were present still conferred the efficacy.

Treatment with celecoxib, a selective COX-2 inhibitor, reduces formation of premalignant adenomatous polyps in the gastrointestinal tracts of humans and mice. In addition to its chemopreventive activity, celecoxib can exhibit antimicrobial activity. Differing bacterial profiles have been found in feces from colon cancer patients compared with those of normal subjects. Moreover, preclinical studies suggest that bacteria can modulate intestinal tumorigenesis by secreting specific metabolites. In the current study, we determined whether celecoxib treatment altered the luminal microbiota and metabolome in association with reducing intestinal polyp burden in mice. Administration of celecoxib for 10 weeks markedly reduced intestinal polyp burden in APCMin/+ mice. Treatment with celecoxib also altered select luminal bacterial populations in both APCMin/+ and wild-type mice, including decreased Lactobacillaceae and Bifidobacteriaceae as well as increased Coriobacteriaceae. Metabolomic analysis demonstrated that celecoxib caused a strong reduction in many fecal metabolites linked to carcinogenesis, including glucose, amino acids, nucleotides, and lipids. Ingenuity Pathway Analysis suggested that these changes in metabolites may contribute to reduced cell proliferation. To this end, we showed that celecoxib reduced cell proliferation in the base of normal appearing ileal and colonic crypts of APCMin/+ mice. Consistent with this finding, lineage tracing indicated that celecoxib treatment reduced the rate at which Lgr5-positive stem cells gave rise to differentiated cell types in the crypts. Taken together, these results demonstrate that celecoxib alters the luminal microbiota and metabolome along with reducing epithelial cell proliferation in mice. We hypothesize that these actions contribute to its chemopreventive activity. Cancer Prev Res; 9(9); 721–31. ©2016 AACR.

Supplementary Figure 1. Pathways coverage map of the significantly altered genes with OME 500ppm treatment in the rat colon tumors as identified using Reactome online tool. Supplementary Figure 2. Significantly down regulated Wnt signaling pathway candidates as visualized using Reactome online tool. (A). Wnt ligand biogenesis and trafficking (B). Degradation of beta-catenin by destruction complex. (C). TCF dependant signaling in response to WNT. (D) Beta-catenin independent WNT signaling. Supplementary Figure 3. Significantly down regulated chemokines related to inflammation pathway candidates as visualized using Reactome online tool. (A). Interleukin-6 family signaling. (B) Interleukin-1 family signaling. (C) Other interleukin signaling. Supplementary Figure 4. Significantly down regulated integrin signalling pathway candidates as visualized using Reactome online tool. Supplementary Figure 5. Significantly down regulated candidates of other key signaling pathways as visualized using Reactome online tool. (A) EGFR signaling. (B) MET signaling (C) Aryl-hydrocarbon signalling pathway

Pathway alterations associated with celecoxib-induced metabolite changes in WT mice.

Supplementary Figure Legends

Supplementary Table 1. Effect of oral OME on terminal body weight and various organs. Supplementary Table 2. Analysis of various serum parameters to determine the effect of OME on liver function (AST & ALT), kidney function (BUN & CREA) and mineral absorption (Mg & Ca). Supplementary Table 3. Partial list of significantly altered pathways (TOP 20) identified using Panther Supplementary Table 4. Selected gene related to tumor growth and progression promoting mechanism / pathways with significantly suppressed gene expression in omeprazole 500 ppm rat colon tumors compared to control.

Supplementary Figure S1. Administration of celecoxib alters the luminal metabolome of WT mice. Feces were collected at baseline (0 weeks) then again at 5 and 10 weeks after initiating celecoxib treatment.

Supplementary Figure 2

Data are rank transformed and displayed as color intensity with low levels indicated by green color and high levels indicated by red color.

Supplementary Figure Legends.

Supplementary Tables

Colorectal cancer causes over 53,000 deaths annually in the United States. Its rising incidences worldwide and particularly in young adults is a major concern. Here, we evaluated the efficacy of omeprazole that is clinically approved for treating acid reflux, to enable its repurposing for colorectal cancer prevention. In the azoxymethane-induced rat colorectal cancer model, dietary omeprazole (250 and 500 ppm) was administered at early adenoma stage (8 weeks after azoxymethane) to assess the progression of early lesions to adenocarcinoma. Administration of omeprazole at 250 or 500 ppm doses led to suppression of total colon adenocarcinoma incidence by 15.7% and 32% (P < 0.01), respectively. Importantly, invasive carcinoma incidence was reduced by 59% (P < 0.0005) and 90% (P < 0.0001) in omeprazole-administered rats in a dose-dependent manner. There was also a strong and dose-dependent inhibition in the adenocarcinoma multiplicity in rats exposed to omeprazole. Administration of 250 and 500 ppm omeprazole inhibited total colon adenocarcinoma multiplicity by approximately 49% and approximately 65% (P < 0.0001), respectively. While noninvasive adenocarcinomas multiplicity was suppressed by approximately 34% to approximately 48% (P < 0.02), the invasive carcinomas multiplicity was reduced by approximately 74% to approximately 94% (P < 0.0001) in omeprazole-exposed rats in comparison with the untreated rats. Biomarker analysis results showed a decrease in cell proliferation and anti-apoptotic/pro-survival proteins with an increase in apoptosis. Transcriptome analysis of treated tumors revealed a significant increase in adenocarcinoma inhibitory genes (Olmf4; Spink4) expression and downregulation of progression promoting genes (SerpinA1, MMP21, IL6). In summary, omeprazole showed significant protection against the progression of adenoma to adenocarcinoma. Prevention Relevance: Preventing colon cancer is urgently needed because of its high incidence and mortality rates worldwide. Toward this end, preventive efficacy of omeprazole, a common medication, was evaluated in animal model of colorectal cancer and was found to suppress colonic adenoma progression to carcinoma. These findings warrant its further evaluation in humans.

Pioglitazone is a PPARγ agonist commonly prescribed for the clinical treatment of diabetes. We sought to expand its use to lung cancer prevention in a benzo[a]pyrene (B[a]P) mouse model with direct lung delivery via inhalation. Initially, we conducted inhalational toxicity experiments with 0, 15, 50, 150, and 450 μg/kg body weight/day pioglitazone in 40 A/J mice. We examined the animals for any physical toxicity and bronchoalveolar lavage fluids for inflammatory and cytotoxicity markers. Doses up to and including 450 μg/kg bw/d failed to demonstrate toxicity with aerosol pioglitazone. For chemoprevention experiments, A/J mice were randomized to treatment groups of inhaled doses of 0, 50, 150, or 450 μg/kg bw/d pioglitazone 1 or 8 weeks after the last dose of B[a]P. For the early treatment group, we found up to 32% decrease in lung adenoma formation with 450 μg/kg bw/d pioglitazone. We repeated the treatments in a second late-stage experiment and found up to 44% decreases in lung adenoma formation in doses of pioglitazone of 150 and 450 μg/kg bw/day. Both the early- and the late-stage experiments demonstrated biologically relevant and statistically significant decreases in adenoma formation. We conclude that aerosol pioglitazone is well-tolerated in the A/J mouse model and a promising chemoprevention agent for the lower respiratory tract. Cancer Prev Res; 10(2); 124–32. ©2016 AACR.

Daily dosing of either NSAIDs or EGFR inhibitors has been shown to prevent bladder cancer development in a N-butyl-(4-hydroxybutyl)nitrosamine (OH-BBN)-induced rat model. However, these inhibitors cause gastrointestinal ulceration and acneiform rash, respectively, limiting their continuous use in a clinical prevention setting. We studied chemopreventive efficacy of pulsatile dosing of EGFR inhibitor erlotinib (42 mg/kg BW, once/week) combined with intermittent or continuous low doses of the NSAID naproxen (30 mg/kg BW/day, 3 weeks on/off or 128 ppm daily in diet) in the OH-BBN induced rat bladder cancer model. The interventions were started either at 1 or 4 weeks (early intervention) or 3 months (delayed intervention) after the last OH-BBN treatment, by which time the rats had developed microscopic bladder lesions. All combination regimens tested as early versus late intervention led to the reduction of the average bladder tumor weights (54%–82%; P < 0.01 to P < 0.0001), a decrease in tumor multiplicity (65%–85%; P < 0.01 to P < 0.0001), and a decrease in the number of rats with large palpable tumors (>200 mg; 83%–90%; P < 0.01 to P < 0.0001). Levels of signal transduction markers, Ki-67, cyclin D1, IL1β, pSTAT3, and pERK, were significantly (P < 0.05 to P < 0.001) reduced in the treated tumors, demonstrating their potential utility as predictive markers for efficacy. These findings demonstrate that significant chemopreventive efficacy could be achieved with alternative intervention regimens designed to reduce the toxicity of agents, and that starting erlotinib and/or naproxen treatments at the time microscopic tumors were present still conferred the efficacy.

The PREVENT Cancer Preclinical Drug Development Program (PREVENT) is a National Cancer Institute, Division of Cancer Prevention (NCI, DCP)-supported program whose primary goal is to bring new cancer preventive interventions (small molecules and vaccines) and biomarkers through preclinical development towards clinical trials by creating partnerships between the public sector (e.g., academia, industry) and DCP. PREVENT has a formalized structure for moving interventions forward in the prevention pipeline using a stage-gate process with go/no go decision points along the critical path for development. This review describes the structure of the program, its focus areas, and provides examples of projects currently in the pipeline.

285 Background: Due to its high prevalence and often indolent natural history, prostate cancer(PC) active surveillance(AS) is an ideal setting for chemoprevention. Studies assessing pomegranate and its extracts have shown promising anti-proliferative and pro-apoptotic effects in cell lines and animal models and a single-arm clinical trial of pomegranate fruit extract(PFE) reported an increase in PSA doubling time(PSADT) during AS. The primary objective of this trial was to assess the effect of PFE supplementation on plasma levels of Insulin-like Growth Factor-1(IGF-1). Secondary objectives addressed PSA doubling time(PSADT), tumor volume on end-of-study(EOS) biopsy and plasma and prostate tissue biomarkers. Methods: Men with organ-confined favorable-risk PC on AS were randomly assigned to receive PFE 1,000 mg(n=15) or placebo(n=15) once daily for twelve months. Prostate biopsies were performed at study entry and upon completion of the one-year intervention. Tissue biomarkers were assessed by immunohistochemistry(IHC) with automated quantitation. Results: PFE was well-tolerated with no significant toxicities. One patient withdrew before study initiation and 29 completed the one-year intervention. No differences in plasma IGF-1 levels(p=0.5), PSADT, or tissue biomarkers of apoptosis or proliferation were observed. A significant increase in urolithin A(a urinary metabolite of pomegranate) was observed in the PFE arm. IHC analyses of both tumor (Table) and normal-appearing tissue adjacent to tumor showed reductions from baseline in IGF-1, 8-OHdG(DNA damage marker), and androgen receptor expression associated with PFE treatment. A trend towards a reduction in the maximum percent of biopsy core tumor involvement was observed(p=0.06) in PFE. Conclusions: PFE administration for 12-months was not associated with a decrease in plasma IGF-1 levels nor an increase in PSADT. However, exploratory analyses suggest that PFE may contain bioactive compounds capable of altering biomarkers in PC and normal-appearing adjacent tissue providing a rationale for further investigation of PFE in the active surveillance population.

Treatment with celecoxib, a selective COX-2 inhibitor, reduces formation of premalignant adenomatous polyps in the gastrointestinal tracts of humans and mice. In addition to its chemopreventive activity, celecoxib can exhibit antimicrobial activity. Differing bacterial profiles have been found in feces from colon cancer patients compared with those of normal subjects. Moreover, preclinical studies suggest that bacteria can modulate intestinal tumorigenesis by secreting specific metabolites. In the current study, we determined whether celecoxib treatment altered the luminal microbiota and metabolome in association with reducing intestinal polyp burden in mice. Administration of celecoxib for 10 weeks markedly reduced intestinal polyp burden in APCMin/+ mice. Treatment with celecoxib also altered select luminal bacterial populations in both APCMin/+ and wild-type mice, including decreased Lactobacillaceae and Bifidobacteriaceae as well as increased Coriobacteriaceae. Metabolomic analysis demonstrated that celecoxib caused a strong reduction in many fecal metabolites linked to carcinogenesis, including glucose, amino acids, nucleotides, and lipids. Ingenuity Pathway Analysis suggested that these changes in metabolites may contribute to reduced cell proliferation. To this end, we showed that celecoxib reduced cell proliferation in the base of normal appearing ileal and colonic crypts of APCMin/+ mice. Consistent with this finding, lineage tracing indicated that celecoxib treatment reduced the rate at which Lgr5-positive stem cells gave rise to differentiated cell types in the crypts. Taken together, these results demonstrate that celecoxib alters the luminal microbiota and metabolome along with reducing epithelial cell proliferation in mice. We hypothesize that these actions contribute to its chemopreventive activity. Cancer Prev Res; 9(9); 721–31. ©2016 AACR.

Colorectal Cancer (CRC) is a major public health issue world-wide with an estimated 700,000 deaths annually. Adults 50 to 69 years of age considered high-risk for CRC can take low-dose Aspirin daily for at least 10 years to reduce their risk for CRC, according to a set of recommendations from the US Preventive Services Task Force. Naproxen is a highly efficacious CRC chemopreventive agent in animal models. Continuous/chronic usage of both drugs is limited by GI toxicity and unwanted side effects. Thus, the rationale to establish intermittent dosing regimens of Naproxen and Aspirin may provide efficacy without GI toxicity. Male F344 rats were used to establish Naproxen and Aspirin pharmacodynamic efficacy and dose-response effects. Rat (36 animals/group) colon cancers were induced by two weekly doses of azoxymethane (AOM). At the adenoma stage, rats were fed diets containing Naproxen (200 and 400 ppm) or Aspirin (700, and 1,400 ppm) either continuously, 1 week on/1 week off, or 3 weeks on/3 weeks off, or Aspirin (2,800 ppm) 3 weeks on/3 weeks off. All rats were euthanized 48 weeks after AOM treatment and assessed for efficacy, dose-response effects, and biomarkers in tumor tissues. Dietary administration of Naproxen and Aspirin did not show any overt-toxicities. Administration of 200 and 400 ppm of Naproxen inhibited colon adenocarcinoma multiplicity by 54.5% and 70.5% (p88% (p67% (p91% (p Citation Format: Altaf Mohammed, Naveena B. Janakiram, Venkateshwar Madka, Yuting Zhang, Anil Singh, Laura Biddick, Qian Li, Stan Lightfoot, Vernon E. Steele, Ronald Lubet, Mark S. Miller, Chen S. Suen, Shizuko Sei, Chinthalapally V. Rao. Intermittent dosing regimens of naproxen and aspirin inhibit azoxymethane-induced rat colon adenoma progression to adenocarcinoma and carcinoma invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4983.

The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation, differentiation, and homeostasis. ROR(alpha) (NR1F1) and Reverb(alpha) (NR1D1) are two members of this family whose biological functions are largely unknown. In addition, no ligand has been yet identified for these two receptors; therefore, they are referred as orphan receptors. Here, we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat L6 myoblastic cells. Ectopic expression of ROR(alpha)1 in L6 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis. Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice, we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice, which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression. Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level. Furthermore, mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter. Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner. Finally, overexpression of GRIP-1/TIF-2, but not SRC-1, potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments. Together, our results identify Reverb(alpha) as a novel target gene for ROR(alpha).

Ligand-dependent gene transcription mediated by the nuclear receptors involves the recruitment of transcriptional coactivators to the ligand-binding domain (LBD), which leads to interaction with the basal transcription machinery, and ultimately with RNA polymerase II. Although most of these coactivators are ubiquitously expressed, a tissue-selective coactivator, PGC-1, has recently been characterized. Because PGC-1 and the retinoid X receptors (RXRs) possess an overlapping tissue distribution, we investigated whether PGC-1 is a coactivator for the retinoid X receptors. In a transient transfection assay, PGC-1 augments ligand-stimulated RXR transcription. Furthermore, PGC-1 efficiently enhances the RXR element-driven reporter gene transcription by all three RXR isoforms. An immunoprecipitation assay reveals that PGC-1 and RXRalpha interact in vivo. In addition, a glutathione S-transferase pull-down assay showed that this interaction requires the presence of the LXXLL motif of PGC-1. We demonstrate further, in a mammalian two-hybrid assay, that this physical interaction also requires the presence of the AF-2 region of RXR to interact with the LXXLL motif of PGC-1, which is consistent with our protein-protein interaction results. A time-resolved fluorescence assay shows that a peptide within the NR box of PGC-1 is efficiently recruited by a ligand-bound RXRalpha in vitro. Finally, PGC-1 and TIF2 synergistically enhance ligand-activated RXRalpha transcriptional activity. Taken together, these results indicate that PGC-1 is a bona fide coactivator for RXRalpha.

Pancreatic cancer (PC) is an almost uniformly lethal disease with inflammation playing an important role in its progression. Sustained stimulation of purinergic receptor P2X7 drives induction of NLRP inflammasome activation. To understand the role of P2X7 receptor and inflammasome, we performed transcriptomic analysis of p48Cre/+-LSL-KrasG12D/+ mice pancreatic tumors by next generation sequencing. Results showed that P2X7R's key inflammasome components, IL-1β and caspase-1 are highly expressed (p < 0.05) in pancreatic tumors. Hence, to target P2X7R, we tested effects of two P2X7R antagonists, A438079 and AZ10606120, on pancreatic intraepithelial neoplasms (PanINs) and their progression to PC in p48Cre/+-LSL-KrasG12D/+ mice. Following dose optimization studies, for chemoprevention efficacy, six-week-old p48Cre/+-LSL-KrasG12D/+ mice (24–36/group) were fed modified AIN-76A diets containing 0, 50 or 100 ppm A438079 and AZ10606120 for 38 weeks. Pancreata were collected, weighed, and evaluated for PanINs and PDAC. Control diet-fed male mice showed 50% PDAC incidence. Dietary A438079 and AZ10606120 showed 60% PDAC incidence. A marginal increase of PanIN 3 (carcinoma in-situ) was observed in drug-treated mice. Importantly, the carcinoma spread in untreated mice was 24% compared to 43–53% in treatment groups. Reduced survival rates were observed in mice exposed to P2X7R inhibitors. Both drugs showed a decrease in caspase-3, caspase-1, p21 and Cdc25c. Dietary A438079 showed modest inhibition of P2X7R, NLRP3, and IL-33, whereas AZ10606120 had no effects. In summary, targeting the P2X7R pathway by A438079 and AZ10606120 failed to show chemopreventive effects against PC and slightly enhanced PanIN progression to PDAC. Hence, caution is needed while treating high-risk individuals with P2X7R inhibitors.

Colorectal cancer (CRC) is the third most common type of cancer in men and women in US. Anti-inflammatory agents have proven to be most effective in CRC prevention, but are associated with gastrointestinal toxicities. Hence, to reduce the acidic environment and enhance the chemopreventive efficacy of anti-inflammatory agent aspirin, a combination of aspirin with acidity neutralizer omeprazole was used in AOM-induced F344 Rat model of CRC. Rat (36/group) colon cancers were induced by AOM (15mg/Kg body weight) s.c., once a week for two weeks. At adenoma stage (13 wks of age), rats were fed diets containing aspirin (700 or 1400ppm), omeprazole (0, 250 or 500 ppm), or aspirin + omeprazole (700 ppm + 250 ppm; 1,400 ppm + 500 ppm). To identify a molecular predictor of the benefit of aspirin plus omeprazole we performed next-generation sequencing of colon adenocarcinomas (AdCa) treated with aspirin (1400 ppm), omeprazole (500 ppm), aspirin plus omeprazole (1400 plus 500 ppm),conducted whole-transcriptome analysis and correlated the molecular portrait with chemoprevention benefit. Aspirin plus omeprazole enhanced colon AdCa incidence inhibitory effects by 55% (p Citation Format: Naveena B. Janakiram, Altaf Mohammed, Jagan M.R. Patlolla, Yuting Zhang, Laura Biddick, Venkateshwar Madka, Li Qian, Stan Lightfoot, Barbara Dunn, Ronald Lubet, Chen S. Suen, Vernon E. Steele, Chinthalapally V. Rao. NGS Transcriptome analysis of colon adenocarcinomas treated with individual and combination of aspirin and omeprazole [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5260. doi:10.1158/1538-7445.AM2017-5260

Pancreatic cancer (PC) still remains a devastating disease that is almost uniformly lethal. Despite advances in the field of molecular genetics in human pancreatic cancers, targeted therapies has not yet translated to an improved overall survival for patients. Chronic inflammation is hallmark of many cancers, including PC. P2×7R is the most potent of the membrane receptors responsible for inflammasome activation and release of inflammatory cytokines. Sustained stimulation of P2×7R drives induction of NLRP inflammasome activation. To understand the role of P2×7 receptor and inflammasome in pancreatic tumor progression, we carried transcriptomic analysis of LSL-Kras pancreatic tumors by next generation sequencing. Results showed that the P2×7 receptor (∼20-fold); key inflammasome components, IL-1β (∼45-fold), caspase-1 (15-fold), IL-18 (∼35-fold), IL-33 (∼93 folds), TNF-α (∼13-fold) and COX-2 (∼41-fold) are highly expressed (p Citation Format: Altaf Mohammed, Naveena B. Janakiram, Venkateshwar Madka, Gopal Pathuri, Qian Li, Rebekah Ritchie, Laura Biddick, Hannah Kutsche, Yuting Zhang, Anil Singh, Hariprasad Gali, Stan Lightfoot, Vernon E. Steele, Chen S. Suen, Chinthalapally V. Rao. Lack of chemopreventive effects of dietary P2×7R inhibitors against pancreatic cancer in p48Cre/+-LSL-KrasG12D/+ mice. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5255.

Colorectal cancer (CRC) protective effects of NSAIDs and COX-2 inhibitors are well established; however, they are associated with gastrointestinal (GI) bleeding and cardiovascular risk. Mechanistic studies suggest that sparing COX-1/2, prostaglandin (PG)I2 synthase and selectively targeting microsomal PG synthase-1 (mPGES-1) and 5-lipoxygenase (5-LOX) would inhibit PGE2 and Leukotrienes (LTs), respectively. To design and develop selective inhibitors of mPGES-1/5-LOX, we used insilico small molecular docking simulation approaches, and identified LFA-9 as a novel selective dual mPGES-1/5-LOX inhibitor among >20 analogs. Azoxymethane (AOM)-induced rat colonic tumors were utilized to assess inhibitory effects of LFA-9 on mPGES-1 and 5-LOX ex-vivo. At 7.5 μM, LFA-9 inhibited the mPGES-1 and 5-LOX activities by ∼57% and ∼68%, respectively. Maximum tolerable dose (100 -1,600ppm) of LFA-9 in AIN-76A diet on male C57Bl/6 mice were tested and observed that Potential CRC preventive efficacy of LFA-9 was assessed in AOM-rat carcinogenesis with colonic Aberrant Crypt Foci, (ACF) as surrogate marker in male F344 rats and intestinal tumor inhibition in APCMin/+ mice. In rats, colonic ACF were induced by AOM/DSS treatment and two week after the AOM/DSS treatment, LFA-9 (200, 400, and 600ppm) was fed by diet and ACF were evaluated after six-weeks. LFA-9 showed dose-response (P60% (P 80% suppression of polyps with >2mm size. Mice fed 350ppm LFA9 showed colon tumor inhibition of 60% (male) and 90% (female). It is noteworthy that both male and female mice fed 700ppm LFA-9 showed 91% inhibition of colon tumors. LFA-9 showed significant suppression of markers of proliferation and inflammation. Overall, above results suggest LFA-9 as a novel dual mPGES-1/5-LOX inhibitor, a safer agent than other NSAIDs and has a potential for prevention of CRC in high-risk patients. (Grant Support: NCI N01-CN 25001-26). Citation Format: Naveena B. Janakiram, Altaf Mohammed, Gopal Pathuri, Venkateshwar Madka, Rebekah Ritchie, Taylor Bryant, Yuting Zhang, Qian Li, Stan Lightfoot, Hariprasad Gali, Steele E. Vernon, Chen S. Suen, Chinthalapally V. Rao. Chemoprevention of colorectal cancer by LFA-9, a novel dual mPGES-1/5-LOX inhibitor: safer approaches to chemoprevention. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2604.

Finding a cure for type 1 diabetes (T1D) has been elusive. Incretin-based therapies, since their approval, have demonstrated their clinical utilities in type 2 diabetes (T2D). Yet, their potential clinical benefits in T1D remain to be appraised. GLP-1, in addition to its insulinotropic action in alleviating hyperglycemia, possesses beneficial effects in protecting progressive impairment of pancreatic β-cell function, preservation of β-cell mass and suppression of glucagon secretion, gastric emptying and appetite. Preclinical data using incretin-based therapies in diabetic NOD mice demonstrated additional effects including immuno-modulation, anti-inflammation and β-cell regeneration. Thus, data accumulated hold the promise that incretin-based therapies may be effective in delaying the new-onset, halting the further progression, or reversing T1D in subjects with newly diagnosed or long-standing, established disease.

Colorectal Cancer (CRC) is a major public health issue world-wide with an estimated over 600,000 deaths annually. Aspirin and Omeprazole are most commonly used medications. Aspirin has been proven as CRC chemopreventive agent, but its usage is limited by GI toxicity. The rationale to establish the chemopreventive role of Omeprazole has been based on i) acidic tumor environment due to the Warburg-effect, and ii) it may protect from the GI toxicity induced by aspirin. Male F344 rats were used to establish omeprazole and aspirin optimal doses, efficacy, dose-response effects of combinations. Rat (36-42 animals per group) colon cancers were induced by two weekly dose of AOM. At adenoma stage, rats were fed diets containing Omeprazole (0, 250 or 500 ppm), or aspirin + omeprazole (700 ppm + 250 ppm; 1,400 ppm + 500 ppm; 2,800 ppm + 250 ppm, respectively) and a combination dose fed every three weeks on and off. All rats were euthanized 48 weeks after AOM treatment and assessed for efficacy, dose-response effects and various biomarkers in colonic tissues. Dietary administration of test agents alone or combinations did not show any overt-toxicities. Administration of 250 and 500 ppm omeprazole inhibited colon adenocarcinoma incidence by 15.7% and 32% (p90% inhibition of invasive adenocarcinomas. In tumors of rats fed omeprazole alone or in combination with aspirin, there was a significant decrease in markers of proliferation (PCNA and β-catenin) and increase in apoptosis (p53, p21, Annexin V) when compared with CRC of rats fed control diet. Surprisingly, we observed a dramatic decrease in the expression levels of proton pump (K+-, H+- ATPase) markers in colonic tumors as compared to tumor adjacent mucosa. These results suggest that omeprazole tumor inhibitory mechanism may not be dependent on tumor proton pump inhibition. Overall, our results suggest that Omeprazole had significant dose-response efficacy effects on the adenoma progression to adenocarcinomas; particularly invasive carcinomas and combining with aspirin significantly enhanced adenocarcinoma inhibitory effects. {This work was supported by NCI-N01-CN-250026} Citation Format: Altaf Mohammed, Jagan M.R Patlolla, Yuting Zhang, Laura Biddick, Venkateshwar Madka, Qian Li, Stan Lightfoot, Ronald Lubet, Chen S. Suen, Vernon E. Steele, Chinthalapally V. Rao. Omeprazole alone, or in combination with Aspirin inhibits azoxymethane-induced colon adenoma progression to adenocarcinoma and carcinoma invasion in F344 rat model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2820. doi:10.1158/1538-7445.AM2015-2820

Numerous studies have demonstrated an inverse relationship between circulating Vitamin D levels and risk of colorectal cancer. Although calcitriol, the most active form of Vitamin D3, exhibits promising antitumorigenic activity, its use as a chemopreventive agent has been restricted by its hypercalcemic effect at therapeutic doses. Eldecalcitol (1α,25-dihydroxy-2β-(3-hydroxypropyloxy) Vitamin D3 or ED-71), a novel analog of calcitriol, is more potent than calcitriol in stimulating bone modeling and is approved for the treatment of osteoporosis in Japan. The goal of the present study was to assess the activity of ED-71 against colorectal adenomas in a unique mouse model of spontaneous multiple intestinal neoplasia (Apc+/Min-FCCC mice). In a preliminary dose-finding study, male Apc+/Min-FCCC mice (6 weeks old, 6-7/group) were treated with vehicle (medium chain triglyceride (MCT)), calcitriol (0.25 μg/kg body weight (bw)), or ED-71 (0.2, 0.1, 0.05, 0.025, 0.0125 μg/kg bw) every other day by gavage for 6 weeks. Body weight was recorded weekly and mice were euthanized at the end of the study. ED-71 was well tolerated at all doses tested, with no significant difference in body weight observed in treated animals as compared to controls. Drug toxicity was assessed by measuring serum calcium levels by colorimetric assay (BioVision Inc.) and hepatic alkaline phosphatase (ALP) by ELISA (Antibodies-online.com). No significant effect of ED-71 on serum calcium levels and hepatic ALP activity was observed. However, because some concern exists regarding the ability of mice treated with high-dose ED-71 (0.2 μg/kg bw) to maintain their body weight long-term, ED-71 doses of 0.05 and 0.1 μg/kg bw were selected for the long-term efficacy study. Prior to entry on this study, male Apc+/Min-FCCC mice (6 weeks old) were prescreened endoscopically to determine tumor-bearing status. Mice were randomized based on tumor status (tumor-free or tumor-bearing) and body weight to one of four treatment groups: vehicle (MCT), calcitriol (0.25 μg/kg bw), ED-71 (0.05 μg/kg bw) or ED-71 (0.1 μg/kg bw). Drugs were administered by gavage every other day for 14 weeks. At the end of the study, intestinal tissue was examined grossly and tumor multiplicity was recorded. No significant effect of ED-71 on tumor multiplicity was observed in tumor-bearing mice. However, in tumor-free mice (11-13/group), treatment with ED-71 (0.1 μg/kg bw) reduced the incidence of gross tumors by 36% (P = 0.017) and colon tumor multiplicity by 30% as compared to that of controls (63.6% vs. 100%, 1.4 ± 0.4 vs. 2.0 ± 0.4, respectively). These findings suggest that ED-71 is effective in inhibiting colon tumorigenesis when given to mice prior to tumor formation. These promising data provide support for future studies to determine the potential utility of ED-71 for the prevention of colorectal cancer in high-risk subjects. (Supported by NCI HHSN261201200015I) Citation Format: Wen-Chi L. Chang, Esther Kaunga, Harry S. Cooper, Lisa Vanderveer, Jing Peng, Yongchao Zhang, Chen S. Suen, Margie L. Clapper. Effect of ED-71, an analogue of Vitamin D3, on intestinal neoplasia in the Apc+/Min-FCCC mouse model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2806. doi:10.1158/1538-7445.AM2015-2806