Search

Colorectal Cancer (CRC) is the third most common cancer and leading cause of mortality in the US. While the rate of CRC incidence has steadily dropped in the last decades, recent rising trends among young adults are a major concern. Hence, preventing CRC in the general population would aid in reducing cancer mortality, especially in high-risk FAP patients for which there are no FDA approved preventive agents. Here, we evaluated TRAIL-inducing agent ONC201, alone and in combination with NSAID naproxen (NAP) for CRC prevention in PIRC rat model of FAP. PIRC rats (20 female rats/group) are bred and randomized by age into control and intervention groups. Starting at 6 weeks of age, PIRC rats were treated with ONC201 by gavage [25mg/kg; 2X/week (LD) or 50mg/kg; 1X/week (HD)]; NAP 200 ppm in AIN-76A diet [continuously or 1 week ON/OFF]; or the combination of ONC201 and NAP. Control group rats received vehicle only. Colonic polyps’ (CP) development (occurrence, number, and size) was monitored by periodic colonoscopies and upon termination at 32 weeks of age. Colonoscopy data indicated significantly fewer CPs in the HD ONC201, NAP alone dosing regimens, and combination groups. At termination, colonic tumors were harvested and subjected to histopathological evaluation. Colonic tumors were histologically classified as hyperplastic polyps, adenomas, and adenocarcinomas (ADCA). In vehicle treated rats, histological assessment showed multiplicity of 2.4±0.4 (Mean±SEM) hyperplastic polyps; 9.14±0.92 adenomas, and 2.4±0.27 ADCA. A significant decrease in the number of hyperplastic polyps (p90% inhibition of adenomas. Importantly, LD ONC201 inhibited colonic ADAC multiplicity by 78.8% (p Citation Format: Venkateshwar Madka, Gopal Pathuri, Karthikkumar Venkatachalam, Srikanth Chiliveru, Yuting Zhang, Nandini Kumar, Nicole Stratton, Stanley Lightfoot, Mark S. Miller, Shizuko Sei, Chinthalapally V. Rao. Colon cancer preventive efficacy of ONC201 and naproxen alone or in combination in FAP relevant PIRC rat model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB114.

Colorectal cancer (CRC) is the 3rd most common cancer in the US with an estimated 149,500 new cases and 52,980 deaths in 2021. Studies show that Western-style diet-induced obesity promote CRC by modulating gut inflammatory mediators. We found significant increase in IL-23 levels (p30) and its. over expression in colonic tumors (CT) from human and rodent models that highly correlated with, the TCGA gene expression & disease-free survival data. These findings suggested IL-23 as a possible important link between obesity and colon tumorigenesis. The present study was designed to understand the role of IL-23 in colorectal carcinogenesis by using genetic knockout (KO) approach. For this study, Apcmin/+ and IL-23 KO mice were crossbred to generate Apcmin/+ mice with IL-23 in heterozygous or KO conditions. To determine the effect of IL-23 on intestinal tumorigenesis, six-week-old Apcmin/+ mice (N≥15/gender) were grouped by IL-23 genotype i.e., normal (+/+), heterozygous (+/-) and KO (-/-) conditions, then maintained under standard conditions. At 20 weeks of age, all mice were euthanized and intestines were evaluated and compared for tumors incidence and multiplicity. Genetic ablation of IL-23 led to significant suppression of large and small intestinal tumors of APCmin/+ mice in both genders. CT multiplicity of male and female IL-23+/+ Apcmin/+ mice were 1.35±0.16 (Mean±SEM) and 0.61±0.13, respectively. Male and female IL-23+/- Apcmin/+ mice had 77% (0.31+0.15; p Citation Format: Venkateshwar Madka, Srikanth Chiliveru, Gopal Pathuri, Janani Panneerselvam, Rajani Rai, Nicole C. Stratton, Nandini B. Kumar, Yuting Zhang, Chinthalapally V Rao. IL-23 knockdown profoundly suppresses intestinal tumorigenesis in APCmin mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3181.

Background: Colorectal cancers (CRCs) is the third most commonly diagnosed cancer and the second most lethal cancer worldwide. CRCs carry high degrees of genomic instability (GI), which enables cancer evolution and makes prognosis poorer. Thus, GI is an exploitable target for therapy purposes. GI can be caused by gene mutation, alteration in gene expression, epigenetic modification, and other infectious or environmental factors. Several mutated genes involved in Chromosome instability and/or microsatellite instability have been identified. Yet it has not been technically feasible to modify functions of mutated genes (e.g., APC, TP53). From the drug development’ standpoint, inhibition of over-expressed target gene is preferred. However, specific genes involved in GI via expression alterations in CRCs have not been comprehensively identified. The gap in knowledge hinders development of CRC therapies targeting GI in CRC. In a previous study, we developed a data mining strategy (Gene Expression to Copy Number Alterations; “GE-CNA”) and comprehensively identified 1,578 genes that associate with CNA in lung adenocarcinoma, among which 39 were survival-critical (i.e., expression levels correlate with significant differences in patients’ survival). Rationale: Causal genes for GI via expressions have not been comprehensively identified in CRCs. Thus, the mechanistic understanding of GI generation in CRC remains incomplete. Few attempts have been made to understand the organ-specificity of genomic instability in cancer. Main Results: We applied the GE-CNA approach to 592 TCGA CRC datasets, and identified 513 genes whose expression levels associate with CNA. Among these, 27 were survival-critical. Comparison with previous results from lung adenocarcinoma indicated striking differences between lung adenocarcinoma and CRC: (a) overall CNA numbers are higher in lung adenocarcinoma than CRC in all stages, (b) 262 CRC-CNA facilitator genes did not show significant concentration in a specific pathway, and (c) 251 CRC-CNA suppressor genes are concentrated in the following pathways: Interferon Signaling, Antigen Presentation Pathway, Heme Biosynthesis II, Natural Killer Cell Signaling, Retinoic acid Mediated Apoptosis Signaling, JAK/Stat Signaling, Glucocorticoid Receptor Signaling, Heme Biosynthesis from Uroporphyrinogen-III I, and Glutathione Redox Reactions II. Implications: The 27 survival-critical genomic instability genes are potential targets to suppress CRC genomic instability and therefore are targets for CRC drug development. Causal genes for genomic instability via expressions differ among organs. Hence, the “targeting genomic instability and/or aneuploidy” approach will need to be tailored for the specific target organ. Citation Format: Chinthalapally V. Rao, Chao Xu, Yuting Zhang, Adam S. Asch, Hiroshi Y. Yamada. Gene expression to copy number alterations analysis in colorectal cancers: Organ specificity of transcriptomic impact on copy number alterations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5003.

Lung cancer patients are more vulnerable to COVID-19 infection. Treatment of patients with lung cancer during the current COVID-19 pandemic is challenging and development of drugs for COVID-19 and lung cancer is urgently needed. Cathepsin L plays key role in lung cancer progression, invasion and more so, endocytosis of SARS-Cov-2 virus into the lung epithelial cells. Importantly, patients with KrasG12V mutation show agreessive disease and high-resistance to chemotherapy due to over-expression of Cathepsin L. Thus, Cathepsin L is a common target for lung cancer and COVID-19 infection. Chymostatin is a known cathepsin L inhibitor, here we screened it for dual inhibition of COVID-19 Mpro and lung cancer patients with COVID-19. In vitro COVID-19 Mpro fluorometric assay was performed to evaluate its inhibitory efficacy by chymostatin. Chymostatin showed dose-dependent inhibition of COVID-19 Mpro activity with IC50 of 15.81 µM (P Citation Format: Nagendra Sastry Yarla, Gopal Pathuri, Simon Terzyan, Yuting Zhang, Anil Singh, Marcus T. Scotti, Venkateshwar Madka, Chinthalapally V. Rao. Chymostatin, a cathepsin L inhibitor, inhibits lung cancer cell proliferation and COVID-19 Mpro in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB074.

Inflammation is a key hallmark of many cancers and potent target for chemoprevention. Experimental and clinical intervention studies indicate strong cancer preventive efficacy of cyclooxygenase (COX)-2 inhibitors. Their use for chemoprevention is limited due to increased cardiovascular (CV) toxicities. Selective COX-2 inhibition diverts arachidonic acid to the 5-lipoxygenase (5-LOX) pathway resulting in accumulation of prothrombotic leukotrienes while depleting antithrombotic prostaglandin (PG)I2, leading to increased risk of CV events. To overcome side effects, balanced dual inhibitors targeting COX-2/5-LOX or microsomal PGE synthase (mPGES)-1/5-LOX enzymes are being developed for chemoprevention. In this study, the clinically advanced dual COX-2/5-LOX inhibitor, licofelone, and its glycine analogue (LFA-9), the mPGES-1/5-LOX dual inhibitor, were evaluated for colon cancer preventive efficacy in the FAP relevant PIRC rat model. In preclinical dose range finding and preliminary toxicity studies in F344 rats, dietary administration of licofelone Citation Format: Venkateshwar Madka, Nagendra S. Yarla, Gopal Pathuri, Yuting Zhang, Anh Bao, Nicole C. Stratton, Anil Singh, David L. McCormick, Altaf Mohammed, Shizuko Sei, Jennifer Fox, Chinthalapally V. Rao. Colon cancer preventive efficacy of licofelone and its analogue LFA-9 in PIRC rat model of FAP [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB225.

Some retrospective studies suggest an inverse risk-association between the use of angiotensin signaling inhibitors and colorectal cancer (CRC). The angiotensin receptor blocker olmesartan medoxomil (OLM) is commonly prescribed for hypertension treatment. Here we sought to determine the chemopreventive efficacy of OLM using the azoxymethane (AOM)-induced CRC rat model. Male and female F344 rats (n=6/gender/group) were randomized and subcutaneously injected with 20 mg/kg of AOM once weekly for two weeks. One week later, AIN76-A diets containing OLM (0 ppm to 480 ppm) were fed to rats. Six weeks after treatment, rats were euthanized and evaluated for toxicity and aberrant crypt foci (ACF) inhibition. OLM-fed rats, particularly at ≥160 ppm, showed signs of toxicity. There was a modest decrease in the total ACFs without any change in multicrypt ACFs in male rats; however in females there was an increase in multicrypt ACFs as compared to the control diet group. For tumor efficacy evaluation, 5-week-old F344 rats were randomized (n=30/gender/group) and CRC was induced as described above. At the adenoma stage (12-weeks after AOM-treatment), rats were administered OLM (0, 20 and 40 ppm) in the diet for 33 weeks, and colonic tumors were evaluated. In the control group, there was a huge disparity in colonic tumor incidence and multiplicity between genders with ~3-fold less tumors in female rats. Surprisingly, colon tumor incidence was significantly increased (3-fold; p Citation Format: Venkateshwar Madka, Yuting Zhang, Gopal Pathuri, Janani Panneerselvam, Nicole Stratton, Anil Singh, Shizuko Sei, Jennifer Fox, Chinthalapally V. Rao. Hypertension drug Olmesartan medoxomil promotes colonic tumorigenesis in AOM-induced CRC rat model [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 15.

Colorectal cancer (CRC) is a major health problem globally. Preventing CRC in the general population, and especially in high-risk cohorts with colonic polyps, or its recurrence, is a high priority. ONC201, an orally active small molecule with robust antitumor activity, is currently in clinical trials for advanced cancers. The aim of this study was to determine the CRC preventive potential of ONC201 using the Apcmin/+ mouse model for CRC in familial adenomatous polyposis. First, the optimal and non-toxic doses of ONC201 were determined. C57BL/6J mice (n=6/group) were randomized, and five different doses of ONC201 (0-100mg/kg BW) were administered orally twice a week by gavage. After six weeks of treatment, all mice were euthanized, and tissues were analyzed for optimal dose and toxicity. Based on the body weight gain, gross organ weights at termination, blood profiling, plasma liver enzyme profile, and histopathological analysis, it was concluded that there were no toxicities associated with ONC201 within the tested dose range. For efficacy evaluation, Apcmin/+ male and female mice (n≥20) were grouped and ONC201 (0, 25 and 50 mg/kg BW) was administered by gavage, twice weekly for 14 weeks. At termination, intestinal tumors were evaluated for incidence and multiplicity to determine the efficacy of ONC201. Results indicate a strong suppressive effect of ONC201 against colonic and small intestinal (SI) tumor multiplicity in mice of both genders. Colonic tumor incidence was significantly reduced by >50% in ONC201-treated (50 mg/kg BW) male (40% in treated vs. 92% in control; p Citation Format: Chinthalapally V. Rao, Venkateshwar Madka, Yuting Zhang, Gopal Pathuri, Janani Panneerselvam, Nicole Stratton, Niklas K. Finnberg, Howard P. Safran, Jennifer Fox, Shizuko Sei, Elizabeth R. Glaze, Robert Shoemaker, Wafik S. El-Deiry. TRAIL inducing small molecule ONC201 prevents intestinal tumors in FAP mouse model [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 14.

Bladder cancer (BC) is the second most common genitourinary cancer and a leading cause of death globally. Muscle invasive BC (MIBC) has high mortality (>85% patients) leading to death within 2 years of diagnosis, if untreated. Although new treatment options were approved recently, it is still very challenging and most expensive to manage. Preventing BC is highly desirable to reduce recurrence, mortality and improve quality of life. Inactivation of p53 signaling and chronic inflammation are frequent hallmarks of MIBC. In spite of the promising chemopreventive effects of non-steroidal anti-inflammatory agents (NSAIDs), their clinical translation is hampered due to side-effects associated with their chronic intake and higher doses. Here we investigated a combinatorial approach to modulate inflammation with NSAIDs (licofelone or NO-Naproxen) and p53 signaling pathways using CP-31398 (CP) for preventing MIBC bladder in-vivo. Transgenic UPII-SV40T mice developing spontaneous MIBC were generated and fed control or experimental diets containing the licofelone (150ppm), NO-naproxen (300 ppm), CP (150ppm) alone or in combination starting at early tumor stage (6 weeks age). After 34 weeks of agent administration, mice were euthanized and urinary bladders were evaluated. Control diet fed transgenic mice developed high grade, muscle invasive, urothelial transitional cell carcinoma (TCC) leading to 3-5 fold increase in bladder weights (140.2±9.8 mg Vs 27.3±0.8 mg; p Citation Format: Venkateshwar Madka, Yuting Zhang, Nandini Kumar, Gopal Pathuri, Nicole Stratton, Stanley Lightfoot, Adam S. Asch, Vernon E. Steele, Altaf Mohammed, Chinthalapally V. Rao. Pharmacological modulation of inflammation and p53 signaling synergize to prevents muscle invasive bladder cancer in-vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5065.

(a) Chromosome instability (CIN), a type of genomic instability that is mitotic error-driven, has a deep impact on carcinogenesis and cancer recurrence. High CIN is a marker for poor prognosis in various cancers, including lung adenocarcinoma. In the tumor genome, CIN can be indicated with Copy Number Alterations (CNA) as its surrogate indicator. Although a diverse array of genes can cause CIN at the cellular level, genes responsible for tumor CNA remain elusive. In this study, we set out to identify genes whose expression shows correlation with CNA in human lung adenocarcinoma. (b) We employed a novel cross-species in silico analysis. Our previous lung RNAseq results from lung cancer-prone CIN mice indicated 348 misregulated genes (92 up-regulated, 256 down-regulated; 2-fold cutoff, P (c) MMP13 (Matrix Metalloprotease 13) is a protease involved in the breakdown of extracellular matrix. NUF2 (yeast NUF2 Kinetochore Protein homolog) is involved in the mitotic process, the misregulation of which directly leads to CIN. CCL22 (C-C Motif Chemokine Ligand 22), ZNF366 (Zinc Finger Protein 366), and GPR114 (G protein-coupled receptor 114, aka ADGRG5) are generally markers for subsets of immune cells (likely dendritic cells). This result led us to further hypothesize that infiltration of CCL22-, ZNF336-, and/or GPR114-positive immune cells inhibits pre-tumor cells and tumor cells with high CNA. (d) The removal mechanism of cells with genomic instability is poorly understood. This is the first time that candidate markers for the immune cells directly or indirectly involved in removal of CIN cells have been identified. This study may aid in fine-tuning cancer-preventive immunotherapy that targets lung cells with genomic instability. Citation Format: Chinthalapally V. Rao, Mudassir Farooqui, Yuting Zhang, Gaurav Kumar, Janani Panneerselvam, Nagendra Yarla, Adam S. Asch, Hiroshi Y. Yamada. Identification of candidate regulators of genomic instability in human lung adenocarcinoma through a new cross-species in silico analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1199.

Colorectal Cancer (CRC) affects more than 1.35 million people worldwide annually. In the US it is anticipated ~140,000 new cases and 50,000 deaths for the year 2017. In spite of existing preventive measures, recent data suggest an increase in the number of CRC incidences in the younger age groups (30 to 55 years) urging the need for better preventives. In the present study we have studied, Diosgenin against colon cancer in APCmin/+ mice and azoxymethane (AOM)-induced rat CRC models. Diosgenin is a steroid saponin with proven diverse medicinal properties, including anti-diabetic, antiobesity and antiinflammatory, and anticancer properties. APCmin/+ mice, a hereditary polyposis mouse model, were bred in-house and starting at 6 weeks age they were given AIN7-6A diets containing diosgenin (0, 500 and 1,000 ppm) for 14 weeks. At 20-wks of age all mice were euthanized and intestines, both small and large, were analyzed for tumors. In the sporadic CRC model, colon tumors were induced in the male F344 rats by two weekly s.c. injections of AOM (15mg/kg BW) followed by diosgenin administration (0 and 1,000ppm) for 38 weeks. Dietary diosgenin showed significant suppression of colon tumor incidence and multiplicity in both models. In APCmin/+ mice both small intestinal polyps (SIP) and colon tumors (CT) were suppressed in a dose-dependent manner with a significant effect in the high-dose treatment. Diosgenin administered female mice had 33% (25±4.1; Mean±SE, p0.05) with high dose in both male and female mice as compared to their respective controls (1.3±0.20 and 1.0±0.25). Importantly, diosgenin showed a similar inhibitory effect on AOM-induced colon adenocarcinoma incidence and multiplicity in the rat model. Administration of diosgenin significantly suppressed both invasive and non-invasive colon adenocarcinoma incidence by 60% (p Citation Format: Venkateshwar Madka, Jayadev Raju, Gopal Pathuri, Snigdha Smriti, Anh Bao, Yuting Zhang, Nicole Stratton, Mudassir Farooqui, Stanley Lightfoot, Adam S. Asch, Chinthalapally V. Rao. Diosgenin, a naturally occurring steroidal saponin, prevents colon cancer in animal models of hereditary and sporadic CRC [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 1269.

Obesity and inflammation play a vital role in colorectal cancer (CRC). Antiobesity agents may be beneficial for CRC prevention. Celastrol is a triterpene bioactive compound derived from Tripterygium wilfordii (TW) plant, which possesses antiobesity and anti-inflammatory properties. In the present study, we tested celastrol for its intestinal tumor inhibitory efficacy, modulation of intestinal microbiome, induction of UCP-1 in inguinal fat, and inflammation under obese conditions using APCMin/+ mouse model. For efficacy study, six-week-old male and female C57BL/6J-APCMin/+ mice (10 mice/group/gender) were fed high-fat diets (HFD; 60% Kcal fat) containing 0 and 150 ppm celastrol and a group of mice with a low-fat diet (LFD; 10% Kca fat) for 11 weeks. At termination, intestinal tumors were evaluated histologically and serum was assayed for fasting glucose, uric acid, liver enzymes (ALT, AST), triglycerides and cholesterol levels. Untreated and treated intestinal tumors were assayed for apoptosis and inflammatory markers by real-time PCR method. Results suggest that administration of LFD showed lower intestinal tumor formation by 52% (p Stephenson Cancer Center Grant. Citation Format: Naveena B. Janakiram, Venkateshwar Madka, Yuting Zhang, Nicole Stratton, Hima Bezawada, Beth Griesel, Altaf Mohammed, Ann L. Olson, Chinthalapally V. Rao. Celastrol inhibits high fat diet-induced obesity and intestinal tumorigenesis in APCMin/+ mice by modulating gut microbes and inflammation [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 4984.

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.

Pancreatic cancer is an aggressive neoplasm with almost uniform lethality and a 5-year survival rate of 7%. Several overexpressed mucins that impede drug delivery to pancreatic tumors have been therapeutically targeted, but enzymes involved in mucin biosynthesis have yet to be preclinically evaluated as potential targets. We used survival data from human patients with pancreatic cancer, next-generation sequencing of genetically engineered Kras-driven mouse pancreatic tumors and human pancreatic cancer cells to identify the novel core mucin-synthesizing enzyme GCNT3 (core 2 β-1,6 N-acetylglucosaminyltransferase). In mouse pancreatic cancer tumors, GCNT3 upregulation (103-fold; P < 0.0001) was correlated with increased expression of mucins (5 to 87-fold; P < 0.04–0.0003). Aberrant GCNT3 expression was also associated with increased mucin production, aggressive tumorigenesis, and reduced patient survival, and CRISPR-mediated knockout of GCNT3 in pancreatic cancer cells reduced proliferation and spheroid formation. Using in silico small molecular docking simulation approaches, we identified talniflumate as a novel inhibitor that selectively binds to GCNT3. In particular, docking predictions suggested that three notable hydrogen bonds between talniflumate and GCNT3 contribute to a docking affinity of −8.3 kcal/mol. Furthermore, talniflumate alone and in combination with low-dose gefitinib reduced GCNT3 expression, leading to the disrupted production of mucins in vivo and in vitro. Collectively, our findings suggest that targeting mucin biosynthesis through GCNT3 may improve drug responsiveness, warranting further development and investigation in preclinical models of pancreatic tumorigenesis. Cancer Res; 76(7); 1965–74. ©2016 AACR.

Pancreatic cancer (PC) is most fatal and is the fourth leading cause of cancer related deaths in USA with a least five year survival of GCNT3 CRISPR knock out PC cells and talniflumate treated PC cells showed a reduction in cell proliferation and significantly reduced spheroid formation (p Citation Format: Altaf Mohammed, Naveena B. Janakiram, Venkateshwar Madka, Gaurav Kumar, Scott Edgar, Gopal Pathuri, Taylor Bryant, Yuting Zhang, Hariprasad Gali, Yan Daniel Zhao, Stan Lightfoot, Chinthalapally V. Rao. Targeting novel mucin synthesis pathway for pancreatic cancer treatment [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 1159. doi:10.1158/1538-7445.AM2017-1159

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) is a lethal disease, and its management is an ongoing challenge. PC is the fourth leading cause of deaths due to cancer in the United States. It is a highly aggressive cancer that is usually diagnosed at an advanced stage, and has the worst prognosis of any malignancy, with a five year survival of Citation Format: Altaf Mohammed, Naveena B. Janakiram, Venkateshwar Madka, Gaurav Kumar, Scott Edgar, Gopal Pathuri, Taylor Bryant, Hannah Kutsche, Yuting Zhang, Laura Biddick, Hariprasad Gali, Yan Daniel Zhao, Stan Lightfoot, Chinthalapally V. Rao. Disruption of mucin synthesis by targeting GCNT3 inhibits pancreatic cancer progression. [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 2621.

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.

Increased polyamine synthesis and inflammation have long been associated with intraepithelial neoplasia and their progression to malignant tumor growth, including lung cancer. Targeting multiple pathways simultaneously with low-dose combinations may be an effective approach to modulate different pathways and their downstream signaling, which may result in an increased efficacy and reduced side effects than a single-agent high dose strategy. The aim of the present study was to investigate the effects of DFMO (ODC inhibitor) and licofelone, a dual 5-LOX-COX inhibitor, individually and in combination, on 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone NNK-induced lung adenoma and progression to adenocarcinoma in female A/J mice. At 6 weeks of age, mice (25 /group) were fed AIN-76A-modified diet, and one week later, lung tumors were induced with a single intraperitoneal (i.p.) injection of 10 μmol NNK/mouse. Three weeks after the NNK treatment, groups of mice were fed with either control or experimental diets containing DFMO (1500 or 3000 ppm) or Licofelone (200 or 400 ppm) or combination of low doses of DFMO and Licofelone. Mice were killed after 17 or 34 weeks of drug exposure and tumors were evaluated via histopathology and lung tumors were assayed for modification of various biomarkers of proliferation and apoptosis. Results suggest that both DFMO and licofelone showed dose-dependent inhibition of NNK-induced lung adenoma progression to adenocarcinoma. At high dose DFMO and Licofelone showed 46% and 55% of adenocarcinoma inhibition. Importantly, low dose combination of DFMO and licofelone showed more pronounced effects at both 17 or 34 weeks in inhibiting the total adenocarcinomas (adenoma and adenocarcinoma progression) by >65% and somewhat in a synergistic manner as compared to individual low doses of DFMO and licofelone. Combination-treated lung tumors exhibited modulation of ODC pathway key components (Arg1, Oat, Oaz, SRM, SMS, and SAT) along with decreased proliferation (PCNA, cyclin D1 and Cyclin A) and increased expression of p53, p21 and p27 compared to tumors from mice fed with control diet. These data suggest that targeting ODC plus 5-LOX/COX decreases the progression of adenoma to adenocarcinoma. Furthermore, adenoma progression delay by combination of DFMO and Licofelone is associated with decreased tumor invasive markers such as MMTPs and EMT markers. In conclusion, targeting two or more pathways is an effective chemopreventive approach for high-risk lung cancer individual's particularly former tobacco smokers with lung hyperplasia and adenomas. (Supported by Kerley-Cade Chair Endowment and NCI-N01-CN-53300) Citation Format: Gaurav Kumar, Jagan Mohan R. Patolla, Venkateshwar Madka, Altaf Mohammed, Li Qian, Yuting Zhang, Laura Biddick, Anil Singh, Allison Gillaspy, Stanley Lightfoot, Levy Kopelovich, Vernon E. Steele, Chinthalapally V. Rao. Simultaneous targeting of ODC and 5-LOX/COX block the tobacco carcinogen-induced lung adenoma progression to adenocarcinoma in A/J 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 2625.

Bladder cancer is the second most common and a leading cause of death among genitourinary cancers worldwide. Particularly, untreated muscle invasive bladder cancer has high mortality (>85% patients) leading to death within 2 years of diagnosis. Preventing this deadly form is highly imperative since the currently available options to patients with invasive disease remained essentially unchanged and no effective drugs have been approved in past two decades. Several non-steroidal anti-inflammatory agents (NSAIDs) have shown promising chemopreventive activity in many cancers. The common adverse events with NSAIDs, especially gastrointestinal (GI) morbidities, including complications in both upper and lower GI tract drives the need for development of safer agents. To overcome this various nitric oxide (NO)-linked NSAIDs have been synthesized. Here we investigated the NO-releasing Naproxen (NO-Naproxen) with proven anti-inflammatory and GI protecting effects for its efficacy in preventing bladder cancer. Transgenic mouse model (UPII-SV40T; n = 30/group) that develop muscle invasive urothelial tumors were generated, genotyped and fed modified AIN-76A diet containing NO-naproxen (0, 300 and 600 ppm) starting at 6 weeks of age. At 40 weeks age, control (0 ppm) and experimental diet (300 and 600 ppm) fed mice were euthanized and urinary bladders were analyzed. Control diet fed male and female transgenic mice developed high grade, invasive transitional cell carcinoma (TCC) of bladder resulting in significant increase in bladder weights (140.2±9.8 mg; p Citation Format: Venkateshwar Madka, Altaf Mohammed, Qian Li, Yuting Zhang, Stanley Lightfoot, Xue-Ru Wu, Vernon Steele, Levy Kopelovich, Chinthalapally V. Rao. Nitric oxide-releasing naproxen prevents muscle invasive bladder cancer. [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 5241.

(a) Colon cancer is the second most lethal cancer and is predicted to claim 50,310 lives in 2014. Chromosome Instability (CIN) is observed in 80-90% of colon cancers and is thought to contribute to colon cancer progression and recurrence. We developed animal models focusing on mitotic error-induced CIN, Shugoshin-1 (Sgo1) haploinsufficient (-/+) mice, to investigate the impact of CIN on colon cancer development. Sgo1-/+ mice showed altered profiles in colonic lesions and cancer development. In this study, we analyzed signature changes in the colonic transcriptome of Sgo1-/+ mice to investigate underlying molecular events in the altered carcinogenesis profiles. (b) We treated control and Sgo1-/+ mice with colonic carcinogen azoxymethane (AOM), and tracked colon cancer development 12, 24, and 36 weeks after AOM treatments. We performed Next Generation Sequencing (NGS)-based transcriptome analysis at 24 weeks. (c) There were 349 hits with 2-fold expression difference threshold, P (d) Systemic Chromosome Instability resulted in transcriptomic changes in metabolic and proliferation regulators in the colon. We propose that the CIN effect may be countered through manipulation of these pathways. Citation Format: Chinthalapally V. Rao, Saira Sanghera, Yuting Zhang, Laura Biddick, Arun Reddy, Stan Lightfoot, Altaf Mohammed, Wei Dai, Hiroshi Y. Yamada. Systemic Chromosome Instability (CIN) resulted in transcriptomic changes in metabolic and proliferation regulators in colonic mucosal tissue of Sgo1-/+ mice. [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 2881. doi:10.1158/1538-7445.AM2015-2881

Galectins, a family of glycan-binding proteins, interact with cell-surface glycoconjugates and influence tumor progression by triggering a cascade of transmembrane signaling events which influence pathogenesis of cancer or tumor outcome. Despite considerable progress in identifying the involvement of individual galectins in tumor biology, an integrated portrait of the galectin network in different organ-site tumors or its microenvironments is not fully established. To understand the role of each galectin in lung, colon, pancreas and urinary bladder tumors and their relevance as markers of tumor progression; chemoprevention intervention trials was evaluated. Lung adenoma and adenocarcinomas were induced by tobacco specific carcinogen, NNK and colon adenomas and adenocarcinomas by azoxymethane (AOM). Whereas, pancreatic ductal tumors were induced by KrasG12D activation in p48Cre.KrasG12D mice and bladder tumors were induced by SV40 activation in UPII-SV40T mice. Expression profiling of galectins was obtained by whole genome-transcriptome analysis with SOLiD methodology from RNAs of tumors and normally appearing tissues from each organ-site. Further profiling and characterization of each galectin (Gal) was carried by RT-PCR, western blotting, and immunohistochemcial analysis. Data from transcriptome suggest that Gal-1, Gal-3, Gal-4, and Gal-12 are significantly associated with tumor progression. Particularly, Gal-1, Gal-3, and Gal-4 expressions were positively correlated with tumor progression in all the above cancers compared to their respective normal tissues and early lesions. Gal-12 was significantly increased in pancreatic and bladder tumor tissues but not in lung and colonic tumors. To understand prognostic value of galectins for chemoprevention interventions, we tested well-established and highly efficacious agents, difluoromethyl ornithine (DFMO), an inhibitor of ODC), and Licofelone, a dual COX-LOX inhibitor in above organ site cancers. DFMO (up to 2,000 ppm) and Licofelone (up to 500 ppm) was administered in the diet to mice after initiation of preneoplastic lesions or adenomas and continued until malignant tumor formations. As anticipated, both agents significantly inhibited malignant tumor formation. Importantly, intervention of tumor growth with mice fed with chemopreventive agents (DFMO and licofelone) showed a significant decrease in expression of Gal-3 and Gal-4 in tumor tissues and correlated with progression of tumor growth inhibition. Overall, these data provide impetus for further studies to delineate the role of Gal-3 and Gal-4 in various cancers and their usefulness as prognostic markers of chemoprevention interventions. (Supported by Kerley-Cade Chair Endowment and NCI-N01-CN-53300) Citation Format: Gaurav Kumar, Venkateshwar Madka, Altaf Mohammed, Jagan M. r. Patolla, Naveena B. Janakiram, Qian Li, Yuting Zhang, Laura Biddick, Allison Gillaspy, Stanley Lightfoot, Chinthalapally V. Rao. Galectin-3 and -4 as signature markers of lung, colon, pancreatic and bladder tumor progression and chemopreventive interventions. [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 1905. doi:10.1158/1538-7445.AM2015-1905

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

Bladder cancer is the second most common genitourinary cancer worldwide. Despite multidisciplinary treatment advances, muscle-invasive bladder cancer continues to have high mortality. Since dysregulation of p53 and mTOR signaling in urothelium is associated with tumor initiation, growth and invasiveness, we investigated whether up-regulation of TP53 plus inhibition of mTOR additively or synergistically inhibits tumor growth and progression in vivo. Transgenic UPII-SV40T male mice (n = 15/group) were generated, genotyped and fed control or experimental diets containing the mTOR inhibitor rapamycin (8 or 16 ppm) and/or the p53 stabilizing agent CP31398 (150 ppm) starting at 6 weeks of age. After 34 weeks, control diet fed transgenic mice developed urothelium-specific high-grade, invasive transitional cell carcinoma (TCC) with significant increase in bladder weights (140.2 ± 9.8 mg; p70%. The combination of CP31398 with 8 ppm rapamycin decreased tumor weight by ∼83% (16.47 ± 5.4 mg; p71% (p Note: This abstract was not presented at the meeting. Citation Format: Venkateshwar Madka, Altaf Mohammed, Qian Li, Yuting Zhang, Laura Biddick, Jagan M.R. Patlolla, Stanley Lightfoot, Xue-Ru Wu, Vernon Steele, Levy Kopelovich, Chinthalapally V. Rao. Modulation of mTOR and p53 signaling using rapamycin plus CP-31398 inhibits growth and progression of urothelial carcinoma in-vivo. [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 4653. doi:10.1158/1538-7445.AM2015-4653

(a) Colon cancer is the second most lethal cancer and is predicted to claim 50,830 lives in 2013. Chromosome Instability (CIN) is observed in 80-90% of colon cancers and is thought to contribute to colon cancer progression and recurrence. For drug development, animal models that reflect cancer-specific etiology is necessary. However, animal models focusing on CIN have not been validated for colon cancer development and for drug testing. In this study, we set out to validate a mitotic error-induced CIN model mice, Shugoshin1 (Sgo1) haploinsufficient mice, as a colon cancer study model. (b) We treated control and Sgo1-/+ mice with colonic carcinogen Azoxymethane (AOM), and tracked colon cancer development over 12, 24 and 36 weeks after AOM treatments. The three endpoints provided information regarding the time course for colon cancer development. (c) Although we predicted an increase in colon tumors in Sgo1-/+ mice at a later (24 or 36 weeks) endpoint, precancerous lesions the Aberrant Crypt Foci (ACF) did not develop to larger tumors straightforwardly. Unexpectedly, the numbers of gross colon tumors showed no difference (P>0.05). Immunohistological analyses indicated IL6, Bcl2 and p16INK4A were differentially expressed in Sgo1-/+ mice. (d) We propose that formation of ACF in Sgo1-/+ mice are facilitated by IL6-STAT3-SOCS3 oncogenic pathway and Bcl2-anti-apoptotic pathway, yet further development of the ACF to tumors are inhibited by p16INK4A tumor suppressor pathway. Manipulating these pathways would be beneficial for inhibiting development of colon cancer with CIN. The CIN mice may be useful for investigating carcinogenesis in earlier stage and tumor regression in later stage. Citation Format: Chinthalapally V. Rao, Saira Sanghera, Yuting Zhang, Laura Biddick, Stan Lightfoot, Wei Dai, Hiroshi Y. Yamada. Antagonizing pathways leading to differential dynamics in colonic carcinogenesis in azoxymethane (AOM)-treated Sgo1 (Shugoshin1)-haploinsufficient chromosome instability (CIN) model mice. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 93. doi:10.1158/1538-7445.AM2014-93

Urinary bladder cancer is the sixth most commonly diagnosed cancer and significant proportion of the patients diagnosed with bladder cancer have high-grade tumors that are at potential risk for post-surgical recurrence and metastasis, where 5-year survival is very low. Hence it is important to identify and develop regimens that prevent urothelial tumorigenesis. Inflammatory pathways particularly, cyclooxygenase (COX-2) and lipoxygenase (5-LOX) mediated metabolites are known to play important role in bladder tumor progression. Consistent with that epidemiological and clinical observations suggest that use of anti-inflammatory agents is associated with reduced risk for bladder cancer. Here we determined the chemopreventive efficacy of a Licofelone, a dual COX-LOX inhibitor, in a transgenic UPII-SV40T mouse model of bladder transitional cell carcinoma (TCC). Six-week old transgenic UPII-SV40T mice (n=30/group) were fed control (AIN-76A) or experimental diets containing 150 or 300 ppm licofelone for 34 weeks. At 40 weeks of age, all mice were euthanized; urinary bladders were collected to determine urothelial tumor weights and histopathological grading. Results suggest that bladders of the control diet fed transgenic mice weighed 3-5 fold more than that of the wild type mice. However, treatment of transgenic mice with licofelone led to a significant, dose dependent inhibition of the urothelial tumor weights (65.2 - 82.7%, p Citation Format: Venkateshwar Madka, Altaf Mohammed, Qian Li, Yuting Zhang, Laura Biddick, Stanley Lightfoot, Xue-Re Wu, Levy Kopelovich, Vernon Steele, Chinthallapally V. Rao. Licofelone, a dual COX-LOX inhibitor prevents transitional cell carcinoma of urinary bladder in UPII-SV40T transgenic mice. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1244. doi:10.1158/1538-7445.AM2014-1244

Preclinical and clinical studies suggest that statins and polyamine inhibitors suppress growth, invasion and progression of malignant colon tumors. Experiments were designed to evaluate the human clinical relevant doses of Rosuvastatin and Difluromethylornithine (DFMO), individually and in combination, on azoxymethane (AOM)-induced colon adenocarcinomas (AdCa) in male F344 rats. Eight weeks of age, rats (30-36/group) treated with AOM to induce the colonic tumors and nine weeks after the AOM treatment, groups of rats were fed AIN-76A diets containing Rosuvastatin (50 or 100 ppm), DFMO (500 or 1000 ppm), or their low dose combination for an additional 40 weeks. AOM-treated rats fed control diet showed AdCa multiplicity of 3.59 ± 0.48 (Mean ±SEM) and AdCa incidence of 96.6 %. 50 or 100 ppm Rosuvastatin significantly suppressed AdCa multiplicities (29%, p52% (p Citation Format: Naveena B. Janakiram, Altaf Mohammed, Taylor Bryant, Yuting Zhang, Misty Brewer, Ashley Duff, Laura Biddick, Stan Lightfoot, Vernon Steele, Chinthalapally V. Rao. Clinically relevant low dose combination of rosuvastatin and difluoromethylornithine provides effective chemopreventive efficacy against AOM-induced colon cancers in F344 rats . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 186. doi:10.1158/1538-7445.AM2013-186

The United States will have approximately 73,510 new cases of bladder cancer in 2012 with 14,880 deaths. More than 50% of the invasive (lamina and muscularis) bladder tumors have mutations in p53 and/or pRB. p53 appears to play a major role in the etiology of human bladder cancer, particularly in the high grade invasive type. Structural and functional defects in p53 are found in majority of human transitional cell carcinomas (TCC). The high prevalence of bladder cancer and the frequency of tumor recurrence make it an important target for chemoprevention. In UPII-SV40T transgenic mice, induced bladder TCC bears strong resemblance to that of human TCC in phenotype and in the mode of progression. In the present study, we determined the chemopreventive efficacy of a p53 stabilizing agent, CP-31398, using transgenic UPII-SV40T mouse model. After genotyping, six-week old UPII-SV40T mice (n=32/group) were fed control (AIN-76A) or experimental diets containing 150 or 300 ppm CP-31398 for ∼34 weeks. Progression of bladder cancer was monitored by MRI imaging. At 40 weeks of age, all mice were euthanized; urinary bladders were collected to determine weights, tumor incidence and multiplicity; and histopathology. There was a significant increase in bladder weights of transgenic vs. wild type mice (male 140.2 mg vs. 27.3 mg, p Citation Format: Venkateshwar Madka, Yuting Zhang, Qian Li, Altaf Mohammed, Stan Lightfoot, Wu Re Xue, Levy Kopelovich, Chinthalapally V. Rao. p53 stabilizing agent CP-31398 prevents urinary bladder tumor growth and invasion in transgenic UPII-SV40T mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3690. doi:10.1158/1538-7445.AM2013-3690

Abstract Mitotic errors have been suspected to be a cause of aneuploidy, rapid accumulation of mutations, and tumorigenesis. In addition, recent cell biological studies demonstrated that mitotic errors can result in DNA damage, which would also aid accumulation of mutations then carcinogenesis. However, no observations in vivo have been provided to support the cell biological observations to date. Previously we developed two strains of high Chromosome Instability (CIN) model mice, which show an increase in mitotic errors due to haploinsufficiency in chromosome cohesion system component Sgo1 or mitotic spindle checkpoint component BubR1. The pathways affected by these mutations are suggested to play a role in aging and carcinogenesis in human. Consistently, compared with wild type, both mice strains showed a significant risk for tumor development in colon with chemical carcinogenesis assays. In this study, we used these CIN mice to test whether (i) spontaneous carcinogenesis occurs in the mice, (ii) spontaneous DNA damage occurs in the mice, (iii) whether associated marker expressions are altered, and (iv) if so, whether it is affected by the age. In 12 month old (middle-age) SGO1 and BUBR1 mice, we observed an increase in spontaneous epithelial cancers, although the carcinogenic effect is not aggressive (p Citation Format: Hiroshi Y. Yamada, Yuting Zhang, Wei Dai, Chinthalapally V. Rao. Consequences of high Chromosome Instability (CIN) in SGO1(Shugoshin1) CIN model mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1752. doi:10.1158/1538-7445.AM2013-1752

Epigenetic modulators particularly histone deacetylases (HDACs) are valid targets cancer for prevention and Therapy. Recent studies from our laboratory and others suggest that HDAC-2 over-expression is associated with colon tumor progression and potential target for colon cancer prevention. The present study explored the chemopreventive effects of novel HDAC-2 inhibitor, OSU-HDAC42, in a mouse model of familial adenomatous polyposis (FAP). We used a rat colon carcinogenesis model assay, inhibition of aberrant crypt foci (ACF) development, as measure of the dose-response and chemopreventive effects. Colonic ACF were induced by azoxymethane (AOM) (15 mg/kg BW once weekly for 2 weeks by s.c. injection at the age of 8 and 9 weeks). One week after AOM-treatment groups of rats were fed AIN-76A diet containing 0, 75, 150 and 300 ppm OSU-HDAC42 for eight-weeks, and then colonic ACF were evaluated. Six week-old male C57Bl/6J-APCmin/+ and wild type mice were fed AIN-76A diet containing 150 ppm OSU-HADC42 or 300 ppm SAHA for 80 days. At ∼120 days of age, all mice were killed, and the intestines were evaluated for polyps. Dietary administration of OSU-HDAC42 produced a dose-dependent inhibition of AOM-induced colonic ACF formation (by 13-50%, p46%, p26%, P≥0.05) in APCmin/+ mice. Whereas 300 ppm SAHA showed limited non-significant inhibition of intestinal polyp formation in APCmin/+ mice. Further, OSU-HDAC42 showed inhibition in small intestinal polyps size measuring 1-2-mm (P Citation Format: Durgadevi Ravillah, Li Qian, Misty Brewer, Yuting Zhang, Laura Biddick, Venkateshwar Madka, Jagan M.R. Patlolla, Altaf Mohammed, Vernon Steele, Chintalapally V. Rao. Targeting HADC-2: Chemopreventive effect of OSU-HDAC42 on rat colon carcinogenesis and APCmin/+ Mice intestinal tumorigenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4392. doi:10.1158/1538-7445.AM2013-4392

CRC is one of the most commonly diagnosed cancers in the United States. Curcumin, a principle compound in turmeric, has been extensively investigated and proven to be antiinflammatory and anticarcinogenic against numerous cancers including CRC. Turmeric is consumed as a dietary ingredient by large populations and turmeric extracts as a dietary supplement by healthy as well as high-risk individuals with various chronic diseases. Turmeric, however, contains over 300 different compounds including essential oils; curcuminoids, turmerones, etc. Most turmeric extracts commonly used as dietary supplements contains Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-177. doi:1538-7445.AM2012-LB-177

Lung cancer is a leading cause of cancer deaths worldwide. p53 Mutations including wild-type p53 nuclear localization due to excessive endogenous electrophilic molecules are highly prevalent in tobacco associated lung cancers. Here, we report the chemopreventive effects of direct-acting p53 modulating agents, CP-31398 and Prima-1 on 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung adenoma and adenocarcinoma in female A/J mice. Seven weeks-old mice on a regular AIN-76A diet, received one dose of 10 μmol NNK /mouse by i.p. injection. Three weeks after NNK treatment, groups of mice (25 mice/group) were fed control AIN-76A or experimental diets containing 50 and 100 ppm CP-31398, or 150 and 300 ppm Prima-1. Mice were sacrificed after 20 weeks (10 mice/group) and/or 36 weeks (15 mice/group) following NNK-treatment and evaluated for lung adenomas and adenocarcinomas by histopathological methods. Administration of 50 and 100 ppm CP-31398 significantly suppressed lung tumor formation by 29% and 39.5% (P Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-176. doi:1538-7445.AM2012-LB-176

Estrogen Receptor (ER)-α signaling is positively associated with colon carcinogenesis as evidenced from pre-clinical and clinical trials. Whereas, down regulation or loss of function of retinoid X receptors (RXR)-α is identified in colon tumors. Both of these receptors play a vital role in growth, development and apoptosis of cells. Bexarotene, a selective agonist for RXR-agonist, is well studied for small cell lung carcinoma and breast cancer. Among the SERMs, raloxifene is an estrogen antagonist in the colon, which has been protective against the colonic lesion development. Hence, experiments were designed to evaluate the chemopreventive efficacies of raloxifene and bexarotene, which were administered individually and in combination during promotion and progression stages of colon tumor formation in F344 rats. Male F344 rats (36-45 animals/group) were fed AIN-76A diet and colon tumors were induced with azoxymethane (AOM; 1.5m/kg body wt). Nine weeks after the AOM-treatment, groups of rats were fed diets containing either raloxifene (1.5 and 3 ppm), bexarotene (50 and 100 ppm), or their low dose combinations for 40 weeks. We have evaluated colonic tumors histopathologically. Also, we have analyzed modulating effects of raloxifene, and bexarotene and combinations of these agents on the markers of colon tumor cell proliferation and apoptosis. Administration of raloxifene at both the doses significantly suppressed (p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1633. doi:1538-7445.AM2012-1633

Lung cancer is the leading causes of cancer deaths. β-Escin a triterpene saponin isolated from horse chestnut (Aesculus hippocastanum) seeds, had tested on 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung adenoma and adenocarcinoma formation in female A/J mice and its possible mode of action was evaluated using human lung cancer cell lines (H460 and H2126). For bioassay, at 6 weeks of age, mice were randomized and fed control AIN-76A modified diet. At 7 weeks of age (25 mice/group) intended for carcinogen treatment received one dose of 10 μmol NNK /mouse by i.p. injection. Three weeks after the NNK treatment, groups of mice fed either control or experimental diets containing 500 ppm β-Escin. Mice were continued on the respective diets and killed at 20 weeks (10 mice/group) and/or 36 weeks (15 mice/group) after NNK-treatment for lung tumor evaluation by histopathological methods. Administration of 500 ppm β-Escin significantly suppressed lung tumor (adenoma+adenocarcinoma) formation by >40% (p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-175. doi:1538-7445.AM2012-LB-175

Inactivation of Shugoshin-1 (Sgo1) function or depletion of Sgo1 in mammalian cells causes missegregation of chromosome. Sgo1 plays an obligate role in chromosome cohesion and its defect is implicated in high chromosome instability. Recent studies indicate that Sgo1 levels were significantly decreased in human colonic tumors, suggesting a role in colon carcinogenesis. Here, we report for first time that depletion of Sgo1 lead to the enhanced colorectal carcinogenesis in mice. To understand the role of Sgo1 in colorectal cancer, we have established haploinsufficient Sgo1+/- mouse model. We tested whether Sgo1 deficiency would enhance the colon carcinogenesis in animal exposed to colonic specific carcinogen. At eight weeks of age mice Sgo1+/+ and Sgo1+/- were treated with azoxymethane (AOM), s.c., 4 mg/kg BW, twice weekly for four weeks. Upon treatment with AOM, we analyzed for colonic crypt cell-cycle modifications and later stage colonic ACF and tumor formation. Sgo1-/+ mice showed higher acute cell death rate than Sgo1+/+ mice soon after the AOM-treatment. However, at the endpoint, the number of aberrant crypt foci [Sgo1+/+, 4±1.41 vs Sgo1+/- 12.2 ±1.5, mean ±SD, p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2983. doi:10.1158/1538-7445.AM2011-2983

Different mode of action exerted by low-dose combination of agents represents a practical approach for improving the chemopreventive efficacy and minimizing unwanted toxicities. Recent clinical studies in high-risk sporadic colorectal cancer patients proven that DFMO combined with COX-1/COX-2 inhibitor, sulindac dramatically reduced polyps formation. Present study designed to address whether i) low-dose combinations DFMO and sulindac would provide the efficacy against rat colon adenocarcinomas in rat model representing human sporadic colon cancers; ii) test the advantages of nitric-oxide releasing (NO)-sulindac would provide colon cancer chemopreventive activity alone or combined with DFMO; and iii) understand the molecular changes associated in colon tumor inhibition by these combinations. Seven-week old, male F344 rats (36 rats/group) were fed the control AIN-76A diet and AOM (15 mg/kg b.w., once weekly for 2 weeks, at the age of 8 and 9 weeks) administered by s.c. Eight weeks after the last carcinogen treatment, rats were fed the diets containing 0 ppm, 500 ppm DFMO, 150 ppm Sulindac, 200 ppm NO-Sulindac individually or in combinations for 40 weeks. Rats were sacrificed and colon tumors were evaluated by histopathologically and colonic tissues were assayed for expression levels of COX-2, iNOS, p21WAF1/CIP1, apoptotic markers, and cell proliferation (BrdU) and ODC enzymatic activity. Bioassay results suggest that administration of 500 ppm DFMO and 150 ppm sulindac significantly suppressed total adenocarcinoma multiplicity by 50% (p0.5-mm. Colonic adenocarcinomas harvested from rats fed DFMO, sulindac or combinations showed significant inhibition of COX-2 and iNOS expression and activity compared to control diet fed rat colonic tumors. Also, DFMO and sulindac or combinations showed considerable enhancement of p21WAF1/CIP1, caspase-3 cleavage, down regulation of bcl-xL, bcl-2 and survivin in rat colonic tumors. Rats that were fed with the combination DFMO and sulindac showed reduced tumor cell proliferation (BrdU labeling index by 16-28%, p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 808. doi:10.1158/1538-7445.AM2011-808