bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4620: Somatic mutation profile of gastric cancer cases from the Hispanic population
Authors: Prieto-Sanchez Rodrigo , Sahasrabudhe Ruta Madhusudan , Lott Paul , Bohorquez Mabel , Jairo Suarez Jhon , Mateus Gilbert , Torres Javier , Echeverry Magdalena , Carvajal-Carmona Luis
Published: August 2015

Gastric cancer contributes to a significant health care burden, being the third most common cause of cancer related deaths worldwide. In the US, gastric cancer incidence and mortality in the Hispanic population is more than double compared to Non-Hispanic whites (NHW). However, a majority of the large-scale gastric cancer genome characterization studies that have been carried so far have been largely limited to non-Hispanic (NH) populations, predominantly Non-Hispanic Whites (NHW). This “mono-ethnic” approach is likely to miss driver mutations that are unique or common in Hispanics but rare in NHWs and can lead to future genomic-driven cancer health disparities. Identification of driver mutations is important to understand the molecular basis of tumorigenesis and to develop better preventive and therapeutic strategies in minority groups like Hispanics. Therefore, the purpose of this study was to characterize gastric cancer somatic changes in the understudied Hispanic population. Somatic mutations were identified using whole exome sequencing (WES) of 36 tumor-normal pairs from Hispanic gastric cancer patients. Microsatellite stability tasting was also performed on all the samples and samples were divided into 27 microsatellite stable (MSS) and 9 microsatellite instable (MSI) cases. Interestingly, preliminary analysis of exome sequencing data delineates differences in mutational profile in our study population compared to publish studies in NH. For example, we found that the frequency of TP53, the most commonly mutated gastric cancer driver in NHs, had a significantly lower frequency in our Hispanic sample (33% in MSS tumors and 11% in MSI tumors compared to 50% in MSS and 35% in MSI tumors reported in TCGA dataset). Similar differences were also found in other frequently mutated gastric cancer genes, which suggest that the genetic pathways leading to gastric cancer in Hispanics are likely to be different to the ones involved in NHW tumors. In summary, our sequencing study of Hispanic gastric cancer cases have identified important differences with other populations, are likely to identify novel cancer drivers in this tumor type and would help bridge the gap in heath care disparities

Citation Format: Rodrigo Prieto-Sanchez, Ruta Madhusudan Sahasrabudhe, Paul Lott, Mabel Bohorquez, Jhon Jairo Suarez, Gilbert Mateus, Javier Torres, Magdalena Echeverry, Luis Carvajal-Carmona. Somatic mutation profile of gastric cancer cases from the Hispanic population. [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 4620. doi:10.1158/1538-7445.AM2015-4620
Abstract 464: Hypoxia-induced miR-210 potentiates MDSC function through regulation of Arg1, Il-16 and Cxcl12, and promotes tumor growth
Authors: Noman Muhammad Zaeem , Chouaib Salem
Published: August 2015

Myeloid Derived Suppressor Cells (MDSCs) are an integral component of the hypoxic tumor microenvironment. We demonstrated that hypoxia selectively induced miR-210 via HIF-1α in splenic MDSCs from tumor-bearing mice. Overexpression of miR-210 enhanced MDSC-mediated T cell suppression under normoxia, while targeting of hypoxic miR-210 decreased MDSC function. Our data show that miR-210 modulated MDSCs function by increasing arginase activity and NO production, although it had no effect on ROS or cytokine (IL-6 and IL-10) production and PD-L1 expression. A comprehensive approach using microRNA target predictions tool, transcriptome analysis, and luciferase reporter assay strongly pointed to the regulation of Arg1, Cxcl12 and Il16 at both mRNA and protein levels by miR-210 in splenic MDSCs. In mediating the immunosuppressive effects of miR-210, targeting of Arg1, and blockade of IL-16 and CXCL12, in splenic MDSCs from tumor-bearing mice under normoxia dramatically decreased MDSC-mediated T cell proliferative response and IFN-gamma production. Interestingly, miR-210 overexpressing MDSCs enhanced in vivo their immune suppressive activity resulting in an increase in 4T1 tumor growth. Therefore, the potential therapeutic use of miR-210 inhibitor oligonucleotide alone, or along with HIF-1α inhibitors as adjuvant tool combined to immunotherapeutic approaches, may be beneficial for boosting the immune system in cancer patients.

Citation Format: Muhammad Zaeem Noman, Salem Chouaib. Hypoxia-induced miR-210 potentiates MDSC function through regulation of Arg1, Il-16 and Cxcl12, and promotes tumor growth. [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 464. doi:10.1158/1538-7445.AM2015-464
Abstract 4640: Andrographolide modulates cell cycle, cell migration and tumor growth in prostate cancer
Authors: Forestier-Roman Ingrid , Sánchez Maria , Casillas Joseph , Rohena Krizia , Martínez-Ferrer Magaly
Published: August 2015

Prostate cancer is the most frequently diagnosed non-cutaneous cancer and is the second leading cause of cancer death in American men. Chemopreventive strategies have been tested to prevent the development of different types of cancer including prostate cancer. Andrographolide, a labdane diterpenoid that is the main bioactive component of the medicinal plant Andrographis paniculata, has been reported to have a wide range of biological activities including anticarcinogenic properties. In this study we aim to determine the role of Andrographolide in the progression of prostate cancer using in vitro and in vivo models. Andrographolide significantly inhibited PC3 cell growth at a concentration of 10μM after 24 h of treatment (P < 0.001), and inhibited LNCaP cell growth at a concentration of 15 μM after 24 h of treatment (P < 0.001). Wound healing assays and boyden chamber experiments showed that PC3 prostate cancer cells treated with Andrographolide (25uM) significantly reduced (P<0.001) their migration and invasion. Flow cytometry assays for Annexin V, cell cycle and cell count and viability were performed. Andrographolide (25uM) significantly increased apoptosis, decreased cell population in G1 phase, and reduced count and viability of PC3 cells treated for 48 hours. Immunofluorescence analysis showed that MMP-11 expression levels were significantly decreased and ZO-1 expression was significantly increased in PC3 cells treated with Andrographolide (25uM). Tumor progression was evaluated using a xenograft model in which the anterior prostate lobes of SCID mice were injected with 250,000 22RV1 cells or 500,000 PC3 cells. Andrographolide was administered bi-weekly at concentrations of 10uM and 25uM intraperitoneal injections during 4 weeks (22RV1) or 8 weeks (PC3). Tumor tissue was collected for gross examination, immunohistochemical analysis and gene expression analysis. In vivo results demonstrated that Andrographolide treatment significantly decreased tumor volume when compared with tumors generated in mice treated with vehicle. Our results suggest that the anti-migration and anti-invasion effects of andrographolide may be associated with alterations in the expression of ZO-1 and MMP-11.

Citation Format: Ingrid Forestier-Roman, Maria Sánchez, Joseph Casillas, Krizia Rohena, Magaly Martínez-Ferrer. Andrographolide modulates cell cycle, cell migration and tumor growth in prostate cancer. [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 4640. doi:10.1158/1538-7445.AM2015-4640
Abstract 4642: Quercetin inhibits prostate cancer by modulating molecules involved in apoptosis and cell proliferation
Authors: Ward Ashely B. , Mir Hina , Kapur Neeraj , Singh Shailesh
Published: August 2015

Prostate cancer (PCa) remains the second leading cause of death in men, despite the multimodal options offered in the clinics. Long latency and indolent nature of PCa provides window of opportunity for preventive interventions using natural and synthetic agents. Quercetin, a bioflavonoid abundant in fruits and vegetables, has been reported to inhibit growth in cancer and reduce inflammation in conditions like prostatitis. Considering this, we investigated the chemo-preventive effects of Quercetin on PCa. The effects of Quercetin on apoptosis and associated proteins on both RNA and protein levels were determined using apoptosis assay, real-time qPCR and western blot, respectively. Furthermore, an antibody microarray was performed to observe changes induced by Quercetin on signaling molecules and possible mechanism of cell survival/apoptosis in PCa. PCa cells treated with Quercetin showed decreased cell proliferation in a time and dose dependent manner, while no significant effect was seen on normal prostate cells. Quercetin treatment also significantly induced apoptosis in PCa cells, as compared with untreated controls. Different phosphorylation status of key molecules (Bcl-2, Akt-1 and NF-k B) was observed following Quercetin treatment in PCa cell lines, as compared with untreated controls, which are known to be involved in supporting PCa progression. These results suggest that Quercetin could be a potential chemo-preventative agent capable of slowing down the progression of PCa, by increasing apoptosis and decreasing cell proliferation.

Note: This abstract was not presented at the meeting.

Citation Format: Ashely B. Ward, Hina Mir, Neeraj Kapur, Shailesh Singh. Quercetin inhibits prostate cancer by modulating molecules involved in apoptosis and cell proliferation. [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 4642. doi:10.1158/1538-7445.AM2015-4642
Abstract 4645: Decursin, a coumarin compound, inhibits the growth of human hepatoma cells involving cell cycle arrest, apoptosis, and autophagy
Authors: Kujur Praveen K. , Yim Dongsool , Singh Rana P.
Published: August 2015

Although recent advances have been made for the treatment of liver cancer, it still remains the sixth most diagnosed cancer worldwide and second most cause of mortality related to cancer. Decursin, a naturally occurring coumarin compound isolated from Angelica gigas Nakai is known for its hepatoprotective activity. For the first time we show that decursin has strong cytotoxic and growth inhibitory effects against liver cancer cells. Decursin (1-25 μM) resulted in a dose-dependent inhibition of Hep3B cell proliferation by upto 65-85% (p<0.005-0.001) in 24-48 h. Similarly in HepG2 cells, decursin (25-100 μM) decreased cell number by upto 61-74% (p<0.005-0.001) at 24-48 h. Growth inhibition was not due to the cell death in HepG2 cells, whereas, Hep3B cells showed an increased cell death (9-45%; p<0.001). Decursin enhanced sub-G1 population of cells in Hep3B but not in HepG2. Further, decursin induced G1 arrest at lower doses, however, at the higher doses cell death prevailed over cell cycle arrest. A consistent G2/M phase arrest in HepG2 cells at all the time points, however, at 100 μM, there was an induction of S-phase arrest. Decursin strongly decreased the colony forming efficiency of Hep3B cells. Decursin induced both apoptotic and autophagic cell death. In flow cytometry analyses, an increase in the acidic vesicular organelle development was observed at 24 and 48 h in Hep3B cells. Interestingly, in decursin treated HepG2 cells, autophagic induction occurred much later (48 h). This was further validated by MDC staining. Mechanistic studies for cell cycle regulators showed that decursin mediated G1 arrest could be via an increase in p21/cip1. Concomitantly, we observed reduced levels of Cyclin A and B1 and their respective kinases. PCNA, a marker of proliferation showed decreased in expression. Decursin mediated induction of apoptosis involved cleavage of caspase-3 and PARP and reduction in survivin level. This was accompanied by an enhanced Bax/Bcl-2 ratio. Additionally, a potent decrease in the phosphorylation of IGF-IR and its downstream effector Akt, was observed. Decursin increased Erk1/2 phosphorylation, however, it was not associated with cell growth or survival. Collectively, these findings suggest that decursin strongly inhibits liver cancer growth and proliferation involving induction of cell cycle arrest, apoptosis and autophagy and could be further explored and developed as novel treatment option for the hepatic malignancies.

Citation Format: Praveen K. Kujur, Dongsool Yim, Rana P. Singh. Decursin, a coumarin compound, inhibits the growth of human hepatoma cells involving cell cycle arrest, apoptosis, and autophagy. [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 4645. doi:10.1158/1538-7445.AM2015-4645
Abstract 4648: Differential regulation of estrogen metabolizing CYP1A1 and CYP1B1 enzymes by novel resveratrol analogs in breast cancer cells
Authors: Ronghe Amruta Mukund , Chatterjee Anwesha , Padhye Subhash , Bhat Hari K.
Published: August 2015

Evidence from epidemiological and experimental studies suggest an association between diets rich in phytoestrogens and reduced risk of breast cancer. Resveratrol, a bioflavonoid, found naturally in grapes, fruits and nuts, has been shown to be a potent chemopreventive agent in vitro. However, poor bioavailability, rapid metabolism and poor specificity limit its use as the preferred anticancer agent. In an attempt to improve the anticancer properties of resveratrol, we synthesized several analogs of resveratrol with appropriate chemical modifications. Of the compounds that were screened for anticancer activity towards breast cancer cells, two analogs, namely, HPIMBD and TIMBD showed better potency and selectivity in inhibiting the growth of breast cancer cell lines while they did not significantly affect the growth of non-neoplastic breast epithelial cell lines. In this study, we investigated the ability of resveratrol and resveratrol analogs in the regulation of estrogen metabolizing genes, cytochrome P450 (Cyp) 1A1, 1B1 and their upstream regulator AhR in breast cancer and non-neoplastic breast epithelial cells.

Non-neoplastic breast epithelial cell line MCF-10A and breast cancer cell lines T47D and MDA-MB-231 were treated with resveratrol, HPIMBD or TIMBD. From cells in culture, mRNA and protein was isolated and expression analyses of CYP1A1, CYP1B1 and AhR carried out by PCR and western blot analyses. Unlike resveratrol, HPIMBD and TIMBD significantly induced the mRNA and protein expression levels of CYP1A1 and AhR while they inhibited that of CYP1B1. Also, HPIMBD and TIMBD induced the expression of CYP1A1 in an AhR-dependent fashion while they inhibited CYP1B1 expression of CYP1B1 by a proteosomal degradation pathway. Cyp1A1 preferentially metabolizes estradiol to 2-hydroxyestradiol and Cyp1B1 to 4-hydroxyestradiol. While, 2-hydroxyestradiol and its methylated product are suggested to be chemo-protective, 4-hydroxyestradiol is regarded as a carcinogenic metabolite of estradiol. Taken together, our findings indicate that novel resveratrol analogs HPIMBD and TIMBD differentially regulate the expressions of CYP1A1 and CYP1B1 in breast cancer cells that may have implications in chemoprevention of breast cancer by these analogs.

Note: This abstract was not presented at the meeting.

Citation Format: Amruta Mukund Ronghe, Anwesha Chatterjee, Subhash Padhye, Hari K. Bhat. Differential regulation of estrogen metabolizing CYP1A1 and CYP1B1 enzymes by novel resveratrol analogs in breast cancer cells. [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 4648. doi:10.1158/1538-7445.AM2015-4648
Abstract 4650: Novel resveratrol-analog HPIMBD inhibits breast cancer cell metastasis by reversal of epithelial-mesenchymal transition
Authors: Chatterjee Anwesha , Ronghe Amruta , Padhye Subhash , Bhat Hari K.
Published: August 2015

Epithelial-mesenchymal transition (EMT) is a biological process that mediates a phenotypical change in the polarized epithelial cells to mesenchymal cells. The process of EMT increases a cell's migratory and invasive properties. It is well documented that the process of EMT plays a very critical role in cancer metastasis. The objective of this study was to characterize the role of a novel Resveratrol analog 4-(E)-{4-hydroxyphenylimino)-methylbenzene, 1,2-diol} (HPIMBD) on EMT in human breast cancer cells.

Human breast cancer cells MDA-MB-231 were treated with Resveratrol (Res) and HPIMBD for up to 48 hours. Protein expression levels of E-cadherin, matrix metalloproteases (MMPs), snail, slug, zeb1/2 and nuclear migration of β-catenin were quantified after treatment with Res or HPIMBD and compared with that of the vehicle-treated control cells. mRNA expression levels of β-catenin and wnt-7b were also quantified. Migration MDA-MB-231 cells was determined after treatment with Res or HPIMBD by wound healing assay.

Treatment with HPIMBD significantly up-regulated E-cadherin protein expression levels. The up-regulation of E-cadherin by HPIMBD was associated with suppression of snail, slug and Zeb1 protein levels. HPIMBD significantly down-regulated mRNA expression levels of β-catenin and wnt-7b. Furthermore, HPIMBD restricted nuclear migration of β-catenin. Treatment of breast cancer cells with HPIMBD resulted in down-regulation of MMP3 and MMP9 but not MMP2 protein expression levels. Treatment with HPIMBD also prevented breast cancer cell migration.

Our studies demonstrtae that HPIMBD decreases the metastatic properties of breast cancer cells by inhibition of EMT and down-regulation of wnt/β catenin pathway. These findings suggest that HPIMBD may have a potential in inhibtion of EMT and may be explored as as a potential therapy to prevent breast cancer metastasis.

Note: This abstract was not presented at the meeting.

Citation Format: Anwesha Chatterjee, Amruta Ronghe, Subhash Padhye, Hari K. Bhat. Novel resveratrol-analog HPIMBD inhibits breast cancer cell metastasis by reversal of epithelial-mesenchymal transition. [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 4650. doi:10.1158/1538-7445.AM2015-4650
Abstract 4653: Modulation of mTOR and p53 signaling using rapamycin plus CP-31398 inhibits growth and progression of urothelial carcinoma in-vivo
Authors: Madka Venkateshwar , Mohammed Altaf , Li Qian , Zhang Yuting , Biddick Laura , Patlolla Jagan M.R. , Lightfoot Stanley , Wu Xue-Ru , Steele Vernon , Kopelovich Levy , Rao Chinthalapally V.
Published: August 2015

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; p<0.0001) compared with wild type (27.3 ± 0.8 mg). Rapamycin (8 or 16 ppm) reduced tumor weight by ∼ 67% or 77% and CP31398 decreased it by >70%. The combination of CP31398 with 8 ppm rapamycin decreased tumor weight by ∼83% (16.47 ± 5.4 mg; p<0.0001). Rapamycin (8 or 16 ppm) suppressed tumor invasion by 53% (p<0.005) or 66% (p<0.0005). CP-31398 alone failed to inhibit tumor invasion; however its combination with low-dose rapamycin inhibited tumor invasion >71% (p<0.0001). Molecular analysis of tumors via real-time PCR, IHC and western blotting showed synergistic inhibition by the drug combination of tumor growth and invasion with suppression of mTOR signaling (mTOR, pmTOR, raptor, rictor, Akt, pAkt); induction of p53 expression; and decreased expression of proliferation (PCNA), cell cycle regulators (cyclin D1, cyclin A), pro-survival molecules (Hif1a, Vegf) and androgen receptor. The combination of CP-31398 and rapamycin appears to be a promising approach for preventing urothelial TCC invasion. (Supported in part by NCI-CN53300)

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
Abstract 4658: Resveratrol aspirinate derivatives as novel chemopreventive agents for colon cancer
Authors: Sang Shengmin , Zhu Yingdong , Fu Junsheng , Shurlknight Kelly
Published: August 2015

In more than a century of use, the benefits of aspirin have been exploited in different therapeutic areas, from inflammation to cardiovascular diseases and cancer prevention. However, the major limitation to the use of aspirin is its side effect to cause ulceration and bleeding in the gastrointestinal tract. The present study found that resveratrol, a natural chemopreventive agent from grapes and berries, and its methylated derivatives can react with acetylsalicyloyl chloride to form resveratrol aspirinate (RAS) and related derivatives. Among all the synthesized derivatives, we found that RAS is the most active component and have much stronger growth inhibitory effect on human colon cancer cells (HCT-116 and HT-29) than resveratrol and aspirin along, as well as the combination of resveratrol and aspirin. In addition, we found that RAS can be hydrolyzed to regenerate resveratrol and aspirin in mice and in human colon cancer cells. Therefore they serve as prodrugs of aspirin and resveratrol and its derivatives. Compared to simply combining aspirin and resveratrol, RAS has not only greater growth inhibitory effects on human colon cancer cells, but also can prevent the damage of gastric epithelial cells caused by direct contact between aspirin and gastric mucosa. Further mechanistic studies found that RAS inhibited cell growth through regulating cell cycle arrest via down-regulation of cyclins D1 and E, and inducing cancer cell apoptosis via mitochondrial pathway. In addition, two stable RAS analogues, for the first time, were successfully synthesized to demonstrate the importance of the scaffold composed of resveratrol and aspirin in the anti-cancer activity of RAS. Our results demonstrated for the first time that resveratrol based aspirin derivatives represent a brand-new, safer and improved alternative to aspirin in anti-cancer and anti-inflammatory therapies.

Note: This abstract was not presented at the meeting.

Citation Format: Shengmin Sang, Yingdong Zhu, Junsheng Fu, Kelly Shurlknight. Resveratrol aspirinate derivatives as novel chemopreventive agents for colon cancer. [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 4658. doi:10.1158/1538-7445.AM2015-4658
Abstract 4664: Impact of TP53 mutations, single nucleotide variants and global methylation patterns on pre-menopausal breast cancer risk
Authors: Samuel Nardin , Lemire Mathieu , Novokmet Ana , Hudson Thomas J. , Malkin David
Published: August 2015

INTRODUCTION: Li-Fraumeni Syndrome (LFS) is a cancer predisposition syndrome that confers a high lifetime risk of developing malignancies, including pre-menopausal breast cancer. Over 80% of LFS patients harbor germline mutations of the TP53 tumor suppressor gene.

OBJECTIVE: We sought to determine if the spectrum of TP53 variants, as well as global methylation methylation patterns, correlates with the presence of TP53 mutations or influences cancer risk in pre-menopausal breast cancer patients.

METHODS: Constitutional DNA was obtained from peripheral blood lymphocytes (PBL) of 10 patients with pre-menopausal breast cancer harboring germline TP53 mutations, 10 patients diagnosed with pre-menopausal breast cancer that are non-mutation carriers, 15 TP53 mutation carriers who do not have cancer, and 15 healthy female controls. Genome-wide methylation analysis of PBL-derived DNA was performed using the Illumina 450K array. Sanger sequencing of exons 2-11, and flanking intron regions of the TP53 gene was performed.

RESULTS: The most commonly observed single nucleotide polymorphism (SNP) in TP53 across all samples was at c.215C>G (p.Pro72Arg) in exon 4 (rs1042522). We observed a G allele frequency of 66.0% in our study group. Three additional low-frequency variants in TP53 were observed: the exon 4: c.108G>A (p.Pro36Pro) variant was found in 2 patients, exon 6: c.639A>G (p.Arg213Arg) was found in 9 patients and intron 9: c.993+12T>C was found in 4 patients. Methylation patterns were not associated with a specific TP53 mutation or variant; however, distinct methylation patterns were observed in TP53 mutation carriers when compared to either TP53 wild-type healthy individuals or patients with early-onset breast cancer.

DISCUSSION: This study represents an epigenome-wide analysis and targeted genetic survey of germline TP53 in early-onset, pre-menoupausal breast cancer. As a result of the mutation frequency observed, germline mutations in TP53 may need to be included in the genetic work-up of patients with early-onset breast cancer. Parallel studies of genome-wide methylation demonstrate these patterns may serve as an independent biomarker of early-onset breast cancer risk.

Note: This abstract was not presented at the meeting.

Citation Format: Nardin Samuel, Mathieu Lemire, Ana Novokmet, Thomas J. Hudson, David Malkin. Impact of TP53 mutations, single nucleotide variants and global methylation patterns on pre-menopausal breast cancer risk. [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 4664. doi:10.1158/1538-7445.AM2015-4664
Abstract 4673: Parity and short-term estradiol treatment confers protection against breast cancer by suppressing cell survival and migration
Authors: Arumugam Arunkumar , Subramani Ramadevi , Nandy Sushmita , Do Duy , Lakshmanaswamy Rajkumar
Published: August 2015

Early pregnancy is the only natural phenomenon known to confer protection against breast cancer. We have earlier demonstrated that short-term treatment with pregnancy levels of estradiol (STET) is very effective in preventing mammary carcinogenesis. The underlying mechanisms behind the protective effect of parity and STET remain unclear. In the current study, we attempted to delineate the influence of the mammary gland microenvironment on mammary carcinogenesis among parous and STET compared to age matched virgin rats. Parous, STET, and virgin (control) Sprague Dawley rats were administered N-methyl-N-nitrosourea (50mg/kg BW) at 15 weeks of age. Mammary cancer development was monitored by weekly palpations for 9 months post carcinogen treatment. A subset of rats (n = 4) were euthanized five weeks post carcinogen treatment. Mammary gland samples were surgically excised and used for histological and molecular analysis. Our analysis demonstrated that parity and STET resulted in reduced expression of cell survival markers (pAKT, pmTOR), extracellular matrix mediated signaling proteins (ERK1/2, pFAK, Itgb1) hormonal (ER, PR, GHR) and growth factor receptors (pEGFR, pIGFR1) compared to age matched virgin group. Further, expression of EMT markers and regulators of cell migration were significantly reduced in parous and STET mammary glands. MicroRNA analysis disclosed that both parity and STET differentially regulated the expression of 23 microRNAs. Among these 18 microRNAs were down regulated and 5 microRNAs were up regulated in both parous and STET treatment. In silico analysis revealed that several of the differentially regulated microRNAs are direct regulators of the cell survival and cell migratory proteins. Parity and STET by altering microRNA expression influences cell survival, cell migration and cell death signaling pathways reduces the cancer promoting microenvironment in the mammary gland.

Citation Format: Arunkumar Arumugam, Ramadevi Subramani, Sushmita Nandy, Duy Do, Rajkumar Lakshmanaswamy. Parity and short-term estradiol treatment confers protection against breast cancer by suppressing cell survival and migration. [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 4673. doi:10.1158/1538-7445.AM2015-4673
Abstract 4677: Control of androgen receptor function by the genomic action of the cochaperone Bag-1L
Authors: Cato Laura , Neeb Antje , Adelmant Guillaume , Ficarro Scott , Westerling Thomas , Marto Jarrod A. , Cato Andrew C. , Brown Myles
Published: August 2015

The androgen receptor (AR) is an important determinant of normal and malignant prostate growth and its function is regulated by many factors, including molecular chaperones, cochaperones and coregulators. One such factor is the nuclear-resident cochaperone Bag-1L. Overexpression of Bag-1L and the amplification of its gene have been reported in the hormone-refractory and metastatic stages of prostate cancer and in androgen-independent prostate cancer cells. However the exact mechanism of Bag-1L-mediated regulation of AR action in prostate cancer remains unclear.

Here we show that Bag-1L and AR directly interact with one another through two distinct domains in both proteins. While the N-terminus of Bag-1L binds to the AR BF-3 domain via a conserved GARRPR motif, the C-terminal Hsp70/Hsc70-binding domain (BAG) of Bag-1L binds to the tau5 region in the intrinsically disordered AR AF1. The latter interaction is of particular significance as tau5 is a region crucial for the ligand-independent transcriptional activity of the receptor. Understanding the details of this interaction will therefore provide new opportunities of controlling AR action for therapeutic purposes outside of targeting the LBD.

We have identified the exact amino acids (K231, K232 and K279) in the BAG domain that are required for the interaction with AR. Amino acid substitution of these residues destroy the interaction of Bag-1L and AR but do not affect the binding to Hsp70/Hsc70. We also observed significant reduction in androgen-dependent cell growth when the BAG-domain mutant is overexpressed in prostate cancer cells in culture and in xenograft mouse models. We further tested the consequence of the BAG-domain mutation on AR transactivation by microarray analysis. We observed repressed expression of multiple genes, including the AR coactivator NCoA2. Correspondingly, we observed multiple changes in the Bag-1L interactome when comparing the wild-type with the BAG mutant; this includes the loss of interaction with the heat shock protein Hsp27 for the mutant Bag-1L protein. Combined these results demonstrate the importance of Bag-1L for AR function and the potential of this protein for therapeutic intervention in prostate cancer.

Citation Format: Laura Cato, Antje Neeb, Guillaume Adelmant, Scott Ficarro, Thomas Westerling, Jarrod A. Marto, Andrew C. Cato, Myles Brown. Control of androgen receptor function by the genomic action of the cochaperone Bag-1L. [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 4677. doi:10.1158/1538-7445.AM2015-4677
Abstract 468: Delayed tumor growth in breast cancer is associated with reduced activity of the a2 isoform of vacuolar ATPase
Authors: Katara Gajendra K. , Kulshrestha Arpita , Gilman-Sachs Alice , Beaman Kenneth D.
Published: August 2015

In cancer cells, vacuolar ATPase (V-ATPase), a multi-subunit enzyme, is expressed on the plasma as well as vesicular membranes and critically influences metastatic behavior. The soluble, cleaved N-terminal domain of a2 isoform of V-ATPase (a2NTD) is associated with in vitro induction of both tumor associated macrophages (TAMs) phenotype and activity in macrophages. This activity caused us to further investigate the role of a2NTD in cancer progression. We evaluated the effects of the knock down of a2 isoform (a2V) in tumor cells and the concomitant tumor microenvironment in the 4T-1 mouse breast cancer model. In vitro experiments showed that macrophages co-cultivated with a2V knocked down (sh-a2) tumor cells produced lower amounts of tumorigenic factors and have a reduced ability to suppress T cell activation and proliferation compared to control tumor cells. Furthermore, sh-a2 4T1 or control 4T-1 cells were inoculated into female Balb/c mice to generate mammary tumors. Data analysis showed that tumor growth in mice inoculated with sh-a2 4T-1 cells was delayed up to ten fold compared to control (mean±SD: 1.3 ± 1.15 vs 13.96 ± 4.06, P>0.01) at day 8 after inoculation. The expression of TAM associated molecules like mannose receptor-1 or CD206, IL-10, Arginase-1, matrix metalloproteinase and vascular endothelial growth factor were significantly down regulated in tumors from sh-a2 inoculated mice compared to control (P<0.01). Additionally, a significant reduction in the number of TAMs (F4/80+CD206+: mean = 6) and increase in macrophage with anti-tumor activity (F4/80+CD11c+: mean = 10.4) was also observed in these tumors which contributed towards delay in tumor growth. Further, we successfully deleted a2V gene in mice and its effect on tumor generation and microenvironment is evaluated. These findings demonstrated that in the absence of a2V in tumor cells, the resident TAM population in the tumor microenvironment is altered, which affected in vivo tumor growth. The data show that by involving the host innate immune system, tumor growth can be control through targeting of a2V and other related factors in tumor cells.

Citation Format: Gajendra K. Katara, Arpita Kulshrestha, Alice Gilman-Sachs, Kenneth D. Beaman. Delayed tumor growth in breast cancer is associated with reduced activity of the a2 isoform of vacuolar ATPase. [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 468. doi:10.1158/1538-7445.AM2015-468
Abstract 4690: Next-generation screen for integrative subtyping and target discovery for KRAS-mutant cancer
Authors: Yuan Tina L. , Bagni Rachel , Yi Ming , Amzallag Arnaud , Afghani Shervin , Beam Katie , Burgan William , Fer Nicole , Garvey Leslie , Smith Brian , Waters Andrew , Stephens Robert , Benes Cyril , McCormick Frank
Published: August 2015

Mutations in the small GTPase, KRAS, are found in ∼140,000 new cases of cancer every year in the United States. This heterogeneous class of cancers manifests primarily as adenocarcinomas of the lung, colon and pancreas. These cancers display a wide spectrum of KRAS-dependency and differentially activate downstream effector signaling. The tumors further diverge in their array of co-occurring secondary mutations, expression signatures and KRAS mutant allele. Ultimately, the sole trait these cancers share in common is an obstinate resistance to chemo- and targeted-therapies, making identification of effective treatments an urgent need. To identify treatments for such a heterogeneous class of cancers, we developed a strategy to stratify KRAS-mutant cell lines into subtypes by integrating next-generation RNAi screening and “Omics” database mining. Each subtype is characterized by unique biomarkers and distinct patterns of effector dependency, both of which represent potential targets for personalized therapeutic strategies.

Our RNAi screen systematically evaluates sensitivity to siRNA-mediated knockdown of 40 KRAS effector nodes in a panel of 135 lung, colorectal and pancreatic cancer cell lines. Data is analyzed on the single cell level, through the simultaneous measurement of 5 functional parameters. This single-cell, multi-dimensional approach allows for a comprehensive assessment of cellular homeostasis, with unprecedented depth and dynamic range that allows robust classification of cell lines by similarity. We identify subtypes of KRAS-mutant cell lines that rely on particular effector pathways such as the RAL, RSK, MTOR and autophagy pathways, which are not engaged by all KRAS-mutant cell lines, and thus may represent targets for personalized treatment. We further identify widely shared dependencies such as on the RAF, glycolytic and cell cycle pathways. Through integrative data mining of exome, transcriptome and drug/siRNA sensitivity databases for each KRAS-mutant subtype, we can identify unique biomarkers that will serve to stratify patients in the clinic and recommend personalized treatment strategies.

Citation Format: Tina L. Yuan, Rachel Bagni, Ming Yi, Arnaud Amzallag, Shervin Afghani, Katie Beam, William Burgan, Nicole Fer, Leslie Garvey, Brian Smith, Andrew Waters, Robert Stephens, Cyril Benes, Frank McCormick. Next-generation screen for integrative subtyping and target discovery for KRAS-mutant cancer. [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 4690. doi:10.1158/1538-7445.AM2015-4690
Abstract 470: Nicotine reduces survival via augmentation of paracrine HGF-MET signaling in the pancreatic cancer microenvironment
Authors: Delitto Daniel , Zhang Dongyu , Han Song , Black Brian S. , Knowlton Andrea E. , Vlada Adrian C. , Sarosi George A. , Behrns Kevin E. , Thomas Ryan M. , Lu Xiaomin , Liu Chen , George Thomas J. , Hughes Steven J. , Wallet Shannon M. , Trevino Jose G.
Published: August 2015

Smoking is an established risk factor for the development of pancreatic adenocarcinoma (PC). However, the relationship between smoking and PC tumor biology is incompletely defined. We report reduced overall survival (OS) in PC patients who continued smoking after surgical resection with curative intention (HR 1.93; P = .040). We further demonstrate augmented paracrine signaling via the hepatocyte growth factor (HGF)/c-Met pathway as a result of nicotine exposure in the PC microenvironment. Specifically, HGF, secreted by patient-derived PC tumor associated stroma (TAS), activates the c-Met receptor in PC cells. This paracrine activation subsequently leads to downstream induction of inhibitor of differentiation-1 (Id1) in PC cells, previously established as a mediator of chemoresistance. Further delineation of the signaling pathway demonstrates HGF-induced Id1 expression is abrogated by silencing of c-Met or pharmacologic inhibition. In patient-derived PC xenografts, nicotine treatment augmented tumor growth and metastasis; tumor lysates from nicotine-treated mice demonstrated elevated HGF expression and phospho-Met levels. Additionally, patients with high intratumoral phospho-Met levels exhibited reduced overall survival compared to those with low phospho-Met levels (Median OS 6.1 vs. 15.2 months, respectively; P = .028). Taken together, our data reveal that nicotine promotes the progression of PC via a microenvironment-dependent, paracrine signaling mechanism.

Citation Format: Daniel Delitto, Dongyu Zhang, Song Han, Brian S. Black, Andrea E. Knowlton, Adrian C. Vlada, George A. Sarosi, Kevin E. Behrns, Ryan M. Thomas, Xiaomin Lu, Chen Liu, Thomas J. George, Steven J. Hughes, Shannon M. Wallet, Jose G. Trevino. Nicotine reduces survival via augmentation of paracrine HGF-MET signaling in the pancreatic cancer microenvironment. [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 470. doi:10.1158/1538-7445.AM2015-470
Abstract 4704: Neo-antigen enriched TIL therapy mediates superior tumor eradication in a patient-derived xenograft model of human melanoma
Authors: Kelderman Sander , Bies Laura , Van Buuren Marit M. , Van Rooij Nienke , Haanen John , Kvistborg Pia , Schumacher Ton
Published: August 2015

The mutational load in melanoma is high relative to that in most other human malignancies, resulting in the possible expression of large numbers of patient-specific mutated antigens. This may, in part, explain the immunogenicity of this disease and the high rate of responsiveness to immunotherapeutic strategies such as tumor-infiltrating lymphocyte (TIL) therapy. Indeed, recent data have shown that cytotoxic T-cell reactivity targeting neo antigens may be common in human melanoma TILs. Importantly though, the clinical relevance of neo-antigen specific T cell populations remains uncertain.

To directly address the tumoricidal potential of defined neo-antigen specific T cell populations, we first identified two neo-antigen specific T-cell populations within a bulk melanoma TIL culture by the combination of exome sequencing and MHC multimer-based T cell screens. Subsequently, we generated TIL products that are highly enriched for these neo-antigen reactivities and we compared the anti-tumor activity of these neo-antigen enriched TIL with that of ‘standard’ bulk TIL in NSG mice bearing the autologous tumor. We observed outgrowth of the tumors in mice treated with standard TIL. In contrast, tumors in mice treated with enriched TILs were controlled long-term. Additional experiments showed that this superior activity of neo-antigen enriched TIL was caused by the increased numbers of T cells with a high anti-tumor activity, rather than the depletion of cell populations with inhibitory activity. Furthermore, once tumors eventually recurred these were still recognized in vitro by cells from the initially infused TIL culture.

Together, these preclinical data demonstrate that neo-antigen reactive T cells within bulk TIL cultures form a critical component of anti-tumor reactivity and provide a further basis to target mutated antigens with cancer immunotherapy.

Citation Format: Sander Kelderman, Laura Bies, Marit M. Van Buuren, Nienke Van Rooij, John Haanen, Pia Kvistborg, Ton Schumacher. Neo-antigen enriched TIL therapy mediates superior tumor eradication in a patient-derived xenograft model of human melanoma. [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 4704. doi:10.1158/1538-7445.AM2015-4704
Abstract 4709: Compartmental flexibility in mammalian folate metabolism
Authors: Ducker Gregory S. , Chen Li , Teng Xin , Rabinowitz Joshua D.
Published: August 2015

Background: Folate metabolism occurs in parallel cytosolic and mitochondrial pathways. Increased mitochondrial, but not cytosolic, folate dependent serine catabolism is among the most consistent metabolic alterations in human tumors. This study employs genetics coupled with quantitative metabolic analysis to determine the respective requirements for compartmentalized folate metabolism in proliferating human cells.

Methods: Using CRISPR-Cas9 technology, we created a library of deletion mutant cell lines for enzymes in folate metabolism. Cellular metabolites from these cells were quantified by liquid chromatography mass spectrometry (LC-MS). Compartmental specific folate metabolic fluxes were assayed by labeling experiments using isotopic tracers of serine, glycine and formate.

Results: We show that in HEK 293T cells, as well as in many but not all human cancer cell lines, folate dependent serine catabolism occurs predominantly in the mitochondria. In mutant cell lines deficient for mitochondrial folate genes, the reversal of the direction of cytosolic one-carbon flux enables cell growth, albeit at a reduced rate and with metabolic hallmarks of 10-formyltetrahydrofolate deficiency. Addition of exogenous formate rescued the 10-formyltetrahydrofolate deficiency in all mutants, but the growth defect in only a subset. The growth defect persisted in cell lines mutant for enzymes required for the generation of mitochondrial one-carbon units and folate dependent generation of NAD(P)H but not for the production of formate.

Conclusions: This study finds that folate metabolism is flexible with respect to compartmentalization. Cytosolic folate metabolism can provide sufficient one-carbon flux to fuel cell growth even in the complete absence of mitochondrial activity. High expression of mitochondrial folate genes in cancer may function to meet biosynthetic requirements under the stressful conditions of the tumor or to provide necessary compartmentalized one-carbon units and/or reducing equivalents.

Citation Format: Gregory S. Ducker, Li Chen, Xin Teng, Joshua D. Rabinowitz. Compartmental flexibility in mammalian folate metabolism. [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 4709. doi:10.1158/1538-7445.AM2015-4709