bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4211: Breast cancer risk factors by mode of detection among screened women in the Cancer Prevention Study-II
Authors: Gaudet Mia M. , Deubler Emily , Diver W. Ryan , Puvanesarajah Samantha , Patel Alpa V. , Gansler Ted , Flanders Dana , Sherman Mark , Gapstur Susan
Published: August 2019

Identifying risk factors for women at high risk of symptom-detected breast cancers that were missed by screening would enable targeting of alternative prevention strategies. To identify breast cancer risk factors by mode of detection, we examined these associations in heavily-screened women from the Cancer Prevention Study (CPS)-II Nutrition Cohort. Among 76,406 women followed for a median of 13.8 years, 2,469 screen-detected and 1,189 symptom-detected breast cancer cases were diagnosed. Multivariable-adjusted associations were estimated using joint Cox proportional hazards regression models with person-time calculated contingent on screening. The mean (standard deviation) age at baseline was 62.0 (6.6) years for non-cases, 61.6 (6.1) for screen-detected cases, and 61.5 (6.2) for symptom-detected cases. Women with symptom-detected tumors were diagnosed at a slightly younger age (mean=69.4) and less likely to be diagnosed with a localized tumor (65.3%), than women with a screen-detected tumor (mean age at diagnosis=70.4 and localized tumors=83.3%). Factors associated with higher risks of symptom detected vs. screen detected breast cancer, included: current combined menopausal hormone use (HR=2.04, 95% CI 1.70 - 2.46 vs. HR=1.43, 95% CI 1.25 - 1.63), current estrogen only menopausal hormone use (HR=1.41, 95% CI 1.15 - 1.72 vs. HR=0.94, 95% CI 0.82 - 1.09), and history of benign breast disease (HR=1.89, 95% CI 1.67 - 2.13 vs. HR=1.49, 95% CI 1.36 - 1.62). The opposite pattern of higher risk of screen detected vs. symptom detected breast cancer was observed for greater adult weight gain (HR=2.32, 95% CI 1.91 - 2.82 vs. HR=1.42, 95% CI 1.10 - 1.84) and alcohol intake (HR=1.39, 95% CI 1.24 - 1.64 vs. HR=1.20, 95% CI 0.92 - 1.58). Our results suggest that understanding risk factors for symptom-detected cancers may help identify women who would benefit from more intensive screening to facilitate early detection.

Citation Format: Mia M. Gaudet, Emily Deubler, W. Ryan Diver, Samantha Puvanesarajah, Alpa V. Patel, Ted Gansler, Dana Flanders, Mark Sherman, Susan Gapstur. Breast cancer risk factors by mode of detection among screened women in the Cancer Prevention Study-II [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 4211.
Abstract 4215: Patient and screening characteristics associated with positive lung cancer screening examinations
Authors: Henderson Louise M. , Sites Samantha , Tailor Tina , Bearden Sara C. , Huamani Roger , Throneburg Allison , Nagle Max , Rivera M Patricia
Published: August 2019

Introduction. The likelihood of having a positive low dose computed tomography (LDCT) lung cancer screening (LCS) examination that requires workup may vary according to patients’ characteristics and risk factors. The purpose of this study is to evaluate patient characteristics associated with positive LCS exams.

Methods. We utilized data from 1684 LDCT exams conducted for LCS at five academic and community sites from 2015-2018. During the screening visit, patients were asked to complete a comprehensive questionnaire including socio-demographics, smoking history, occupational and environmental exposures, family history of lung cancer, and comorbid conditions. Information on the LCS examination, including the radiologists’ Lung Reporting and Data System (Lung-RADS) assessment, was obtained from the radiologist report. We dichotomized Lung-RADS into negative (Lung-RADS 1 (negative) or 2 (benign appearance or behavior)) and positive (Lung-RADS 3 (probably benign), 4A (suspicious), or 4B (suspicious)) based on the management recommendation of continuing with annual screening in 12 months or requiring follow-up before 12 months. We compared Lung-RADS results (positive versus negative) by patient characteristics using chi-square tests and examined predictors of positive LCS exams using multivariate logistic regression, reporting adjusted odds ratios (aORs) and 95% confidence intervals (95%CI).

Results. Screened patients ranged in age from 50-78 years, with 46.7% less than 65 years and 53.3% ages 65 or older. Approximately 53.3% were male and 46.7% were female; 84.5% were White, 11.8% were African American, and 3.7% were other races. The majority (76.5%) of screened patients were overweight or obese. Two-thirds (68.5%) of patients had more than one LCS exam. Most (85.0%) LCS exams were negative while 15.0% were positive. There were no significant differences in Lung-RADS assessment by race, gender, education, or body mass index. Lung-RADS were significantly more likely to be positive in those ages 65 and older versus those ages less than 65 (aOR=1.47, 95%CI: 1.03-2.10) and in baseline versus subsequent screening exams (aOR=0.43, 95%CI: 0.30-0.62).

Conclusion. Predictors of having a positive LCS exam requiring work-up before the next recommended annual screening test include being aged 65 and older and having a baseline LCS examination.

Citation Format: Louise M. Henderson, Samantha Sites, Tina Tailor, Sara C. Bearden, Roger Huamani, Allison Throneburg, Max Nagle, M Patricia Rivera. Patient and screening characteristics associated with positive lung cancer screening examinations [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 4215.
Abstract 4236: Evaluating alterations in mRNA biogenesis with TCGA breast cancer sequencing data
Authors: Yan Thomas J. , Burow Matthew E. , Martin Elizabeth C.
Published: August 2019

Alternative splicing leads to the production of multiple mRNA isoforms of a gene transcript with distinct functional characteristics. These events are frequently observed in rapidly dividing cancer cells and are broadly recognized as important signatures for oncogenesis pathways and treatment options. In this study, we utilized data from The Cancer Genome Atlas (TCGA) to identify differences in isoform expression between estrogen receptor-positive and estrogen receptor-negative breast cancer cell lines. RNA sequencing analysis combined with clinical annotation data revealed alterations in isoform expression that were associated with patient survival. These global changes in isoform expression occurred independently of total gene expression in both estrogen receptor-positive and -negative breast cancer. Furthermore, we identified multiple underrepresented isoforms including CDKAL1 and CSTF3 that were associated with favorable prognostic outcomes despite a lack of association observed with their traditional transcripts. These data suggest an underlying role mediated by alternative spliced isoforms in driving clinical outcome in breast cancer, warranting further investigation in delineating isoform expression profiles in previously established cancer pathways.

Citation Format: Thomas J. Yan, Matthew E. Burow, Elizabeth C. Martin. Evaluating alterations in mRNA biogenesis with TCGA breast cancer sequencing data [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 4236.
Abstract 4241: Molecular profile of tumor-associated adipose in obese women with breast cancer
Authors: Infante Niel , Coad James , Primerano Donald , Denvir James , Vona-Davis Linda
Published: August 2019

Breast tumors have a direct physical and vascular interface with white adipose tissue, making them ideal model systems to study the adipose-tumor microenvironment, especially within the context of obesity. As a first initiative to understanding how adipose tissue contributes to breast tumor etiology, we explored cancer-associated adipose tissue in patients with invasive breast cancer. We hypothesized that obesity leads to greater alterations in cancer-associated adipose tissue by expressing genes involved in cellular growth, angiogenesis, and invasion, thus providing support for breast cancer progression. We also proposed that adipocytes adjacent to tumors express a genotype unique to obese breast cancers. Adipose tissue samples (n=10) were obtained from obese and lean women with breast cancer. Within individuals, tissue was sampled proximal and distal (2 cm) to the tumor. Using next-generation whole transcriptome sequencing, we examined genes from cancer-associated adipose tissue in both groups. A heat map of normalized expression values was generated with unsupervised clustering and the fifty most informative genes. Samples from obese individuals with breast cancer tended to cluster together. There were significant gene differences between adipose collected proximal versus distal from the tumor in obese but not in lean women, such as genes associated with myosin heavy chains MYH1, 4, 8, and 11 (adj. p < 0.05). In examining obese women, there were 16 Hallmark pathways that were statistically significant (adj. p < 0.10); whereas a subset of these (11 pathways) were significant in lean women. Of interest, pathways that were significant in both were pathways that regulate EMT, adipogenesis and TGF beta signaling. The molecular signature of cancer-associated adipocytes is different between lean and obese patients. (Supported by NIH P20GM103434 and NIGMS U54GM104942)

Citation Format: Niel Infante, James Coad, Donald Primerano, James Denvir, Linda Vona-Davis. Molecular profile of tumor-associated adipose in obese women with breast cancer [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 4241.
Abstract 4266: Endoplasmic reticulum (ER), a potential therapeutic target for mutant p53 colorectal cancer
Authors: Tyagi Ashish , chandrasekaran Balaji , Kolluru Venkatesh , Baby Becca V. , Houda Alatassi , Ramisetti Srinivasa , Sharma Arun , Ankem Murali , Damodaran Chendil
Published: August 2019

Introduction: Inactivating p53 mutations contribute to tumor progression and treatment-resistance, resulting in poor patient survival in colorectal cancer patients (CRC). The goal of this study is to identify novel small molecules that therapeutically target mutant p53 in CRC.

Methods: We analyzed the effects of small molecule (ASR458) on p53-wild type (p53-wt; HCT116) and mutant p53 (p53-mut; SW620) colon cancer cells by phenotypic, molecular and in vivo assays.

Results: ASR458 treatment significantly inhibited the proliferation of both HCT116 and SW620 cells at nM concentrations. In p53-wt HCT116 cells, ASR458 caused induction of p53 that resulted in caspase-mediated cell death in both in vitro and in vivo models. On the contrary, ASR458 treatment induced ER-stress signaling (i.e., phosphorylation of ERK and eIF2-α) in p53-mut SW620 cells, which triggered ATF4 activation and subsequent induction of cascade of autophagy events (Atg family proteins, LC3B and Lamp1), causing autophagy-mediated cell death. Silencing ER stress marker ATF-4, a key regulator of autophagy, caused resistance to ASR458 and abrogated autophagy signaling in SW620 cells. This suggested that induction of ER-stress is critical for the cytotoxic effects of ASR458 in p53-mut CRC. Preliminary knockdown studies indicate that silencing of ER markers causes resistance to ASR458 in vitro, further confirming ER-stress as the mechanism of ASR458 action in p53-mut CRC. ASR458 significantly inhibited the growth of SW620 tumors in xenograft. Tissue analysis confirmed the ER-stress signaling observed in vitro.

Conclusion: Our results demonstrate ASR458 as a potential therapeutic agent with distinct targets in p53-wt and p53-mut CRC. This study also suggests that ATF4 mediated autophagy in unmanaged ER stress can reduce CRC pathogenesis. Further investigation into the pharmacokinetics and pharmacodynamics of ASR458 should help clinical translation of this agent.

Citation Format: Ashish Tyagi, Balaji chandrasekaran, Venkatesh Kolluru, Becca V. Baby, Alatassi Houda, Srinivasa Ramisetti, Arun Sharma, Murali Ankem, Chendil Damodaran. Endoplasmic reticulum (ER), a potential therapeutic target for mutant p53 colorectal cancer [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 4266.
Abstract 4278: A comparison of drug sensitivity in isogenic tumor cell lines confirms that cytoprotective autophagy confers intrinsic resistance to cisplatin
Authors: Xu Jingwen , Patel Nipa , Saleh Tareq , Wu Yingliang , Lima Santiago , Gewirtz David A.
Published: August 2019

Although autophagy inhibition often increases tumor cell sensitivity to chemotherapy and radiation, these observations do not provide definitive proof that autophagy is actually an intrinsic mechanism of resistance since virtually all tumor cells undergo autophagy in response to therapy. To directly address whether autophagy induction can confer chemoresistance, the current studies utilized a paired set of isogenic cell lines in which (cisplatin-induced) autophagy was either cytoprotective (crp53 cells where p53 was knocked out) or nonprotective (parental p53 wt cells) in function, but where the overall extent of autophagy was essentially identical. Cisplatin sensitivity was significantly lower in the crp53 cells where cisplatin-induced autophagy was protective compared to the parental p53 wt cells where cisplatin-induced autophagy was nonprotective, in support of the premise that cytoprotective autophagy could confer a relative degree of intrinsic resistance to chemotherapy. Accordingly, we can draw the following conclusions: (i) radiation and chemotherapy can promote both protective and nonprotective autophagy in tumor cells; (ii) although p53 can act as an autophagic switch, it cannot be reliably predicted whether autophagy will be protective or nonprotective based solely on p53 function; and (iii) even in the case(s) where autophagy is cytoprotective, it cannot be assumed that autophagy will confer drug or radiation resistance, but only that autophagy inhibition may, in select cases, increase drug or radiation sensitivity. These findings highlight the complexity of efforts to exploit autophagy inhibition as a therapeutic strategy to enhance tumor response to chemotherapy and radiation.

Citation Format: Jingwen Xu, Nipa Patel, Tareq Saleh, Yingliang Wu, Santiago Lima, David A. Gewirtz. A comparison of drug sensitivity in isogenic tumor cell lines confirms that cytoprotective autophagy confers intrinsic resistance to cisplatin [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 4278.
Abstract 4284: Targeting notch signaling in glioblastoma cancer stem cells through modulation of Connexin43 function
Authors: Lunski Michael , Smyth James , Vaughn Jennifer , Sheng Zhi , Gourdie Robert , Purow Benjamin , Lamouille Samy
Published: August 2019

Glioblastoma (GBM) is a highly malignant and lethal cancer of the central nervous system for which there has been limited progress in improving patient outcomes despite intensive research efforts. The current multimodal therapy for newly diagnosed GBM patients includes surgical resection, radiotherapy and cytotoxic chemotherapy with temozolomide (TMZ), conferring a median survival time of just 14.6 months. Failure to generate more effective treatment strategies is due to 1) the infiltrative nature of GBM tumor cells preventing complete surgical resection, and 2) the cellular heterogeneity within GBM tumors, which often comprise a sub-population of GBM cancer stem cells (GSCs) characterized by self-renewal characteristics and resistance to chemotherapeutic alkylating agents including TMZ. Multiple signaling pathways, including Notch, participate to the formation and maintenance of GSCs. Recent studies, including our research, have shown that increased levels of gap junction protein Connexin43 (Cx43) correlate with TMZ resistance in GBM cells and inversely correlated with GBM patient survival. Importantly, Cx43 has also been associated with anti-proliferative effects in glioma, and reduced levels of Cx43 protein occur in high-grade gliomas. Therefore, we hypothesized that altering Cx43 localization and/or activity rather than Cx43 expression represents a potent strategy for GBM treatment. Regulating Cx43 function is primarily associated with the multiple sites for post-translational modifications and protein-protein interactions within the Cx43 carboxy-terminus. Our research identifies crosstalk between Cx43 and Notch signaling in GSCs. Using a Cx43 carboxy-terminus mimetic peptide that modulates non-junctional functions of Cx43, we observe a decrease in Notch1 protein expression and reduced transcription of its downstream targets Hes1 and Hey1 in GSCs derived from patient tumors. Most importantly, our Cx43 mimetic peptide decreases cell survival in TMZ-resistant GSCs, limits GSC neurosphere formation in vitro, and inhibits GSC-derived tumor growth in vivo. Our current research aims at dissecting the molecular mechanisms of Cx43 functions on Notch signaling in GSCs using this Cx43 mimetic peptide. In conclusion, we have identified a novel therapeutic opportunity to decrease the tumorigenic potential of GSCs through altering Cx43 activity and Notch signaling to target chemoresistant GSCs in GBM treatment.

Citation Format: Michael Lunski, James Smyth, Jennifer Vaughn, Zhi Sheng, Robert Gourdie, Benjamin Purow, Samy Lamouille. Targeting notch signaling in glioblastoma cancer stem cells through modulation of Connexin43 function [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 4284.
Abstract 4291: p85α alters Gab2 phosphorylation profile in ovarian cancer
Authors: Li Xinran , Mak Victor CY , Cheung Annie NY , Mills Gordon B. , Cheung Lydia WT
Published: August 2019

Dysregulation of phosphatidylinositol 3-kinase (PI3K), which is essential for various cellular processes, is a common event in cancers. PIK3R1 encodes the PI3K class IA regulatory subunit p85α, which binds the p110 catalytic subunit forming heterodimeric PI3K complex. The PI3K associates with receptor tyrosine kinase, for example through the docking protein Gab2, to initiate downstream signaling events. Downregulation of p85α is frequently seen in cancers. However, the signaling changes driven by the downregulation remain to be elucidated. Our data herein indicate that reduced p85α expression leads to changes in the phosphorylation status of Gab2 at multiple amino acid residues. PIK3R1 siRNA decreased the levels of phosphorylated Y452 (a site implicated in binding p85α) and S210 (a site that may mediate negative feedback of Gab2 activity). Meanwhile, we observed an increase in phosphorylated S623, which was shown to involve in STAT activation. These findings provide mechanistic insights on the pathway activation upon downregulation of p85α in cancer.

Citation Format: Xinran Li, Victor CY Mak, Annie NY Cheung, Gordon B. Mills, Lydia WT Cheung. p85α alters Gab2 phosphorylation profile in ovarian cancer [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 4291.
Abstract 4294: C6 Ceramide potentiates paclitaxel and gemcitabine mediated antitumor effects against pancreatic cancer via inhibition of prosurvival PI3k/AKT/mTOR and ERK pathways
Authors: Wanebo Harold J.
Published: August 2019

The 5% survival of pancreatic cancer emphasizes need for new treatment strategies. Our studies suggest potential value of C6Ceramide (C6Cer) as a chemotherapeutic adjunct. The current study demonstrates that combination chemotherapy of Paclitaxel with adjunct C6Cer significantly increased cell death (apoptosis) in pancreatic cancer cell lines (L3.6,PANC-1, MIA (PaCa-2) in vitro as well as in vivo - enhancing tumor regression in SCID mice bearing L3.6 transplants. A similar synergistic effect was observed with addition of C6Cer to cetuximab which is not effective in KRAS mutant Pancreatic or Colorectal cancer and thus is not used clinically. At the molecular level combining C6Cer with Gemcitabine or Paclitaxel resulted in significant decreased p-AKT and p-GSK-a/b(PI3K/AKT (survival)pathway in L3.6 cells.

Conclusion: C6Cer significantly inhibited key cancer survival pathways resulting in significant synergism with chemotherapeutics in therapy of Pan Ca and KRAS mutant CRC.

Citation Format: Harold J. Wanebo. C6 Ceramide potentiates paclitaxel and gemcitabine mediated antitumor effects against pancreatic cancer via inhibition of prosurvival PI3k/AKT/mTOR and ERK pathways [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 4294.
Abstract 4299: Targeting AKT with oridonin inhibits growth of esophageal squamous cell carcinoma in vitro and patient-derived xenografts in vivo
Authors: Song Mengqiu , Zhao Ran , Xie Hua , Chen Hanyong , Liu Kangdong , Bode Ann M. , Lee Mee-Hyun , Dong Zigang
Published: August 2019

Esophageal cancer is the sixth leading cause of cancer death worldwide and has low survival rates with poor prognosis. Hyperactivation of AKT has been reported to modulate cell growth, survival, and gene expression in various solid tumors. Oridonin, an inflammatory medical and diterpenoid compound isolated from Rabdosia rubescens, has exhibited various pharmacologic and physiologic properties, including antitumor, antibacterial, and anti-inflammatory effects. In this study, we aimed to investigate the effects of oridonin on the proliferation and growth of ESCC and to elucidate its underlying mechanisms of action. We found that oridonin is an inhibitor of AKT and induces cell cycle arrest and apoptosis in ESCC cells and attenuates growth of patient derived xenograft (PDX) tumors in vivo by interfering with AKT signaling pathways through MTT assay, anchorage-independent cell growth assay and Cell cycle and apoptosis analyses. AKT was developed as a dierctly target of oridonin by in vitro kinase assay, ex vivo and in vitro pull-down assay and computational docking model. Mechanistically, oridonin diminished the phosphorylation and activation of AKT and supressed the downstream of p-GSK-3β, p-mTOR and NF-kB activity. Moreover, a combination of oridonin and 5-fluorouracil or cisplatin (clinical chemotherapeutic agents) enhanced the inhibition of ESCC cell growth. Overall, our study suggests that oridonin can inhibit progression of ESCC tumors in vitro and in vivo by suppressing AKT signaling through its direct targeting of AKT. Thus, this study might provide useful information in the clinical application of oridonin for ESCC chemotherapy.

Citation Format: Mengqiu Song, Ran Zhao, Hua Xie, Hanyong Chen, Kangdong Liu, Ann M. Bode, Mee-Hyun Lee, Zigang Dong. Targeting AKT with oridonin inhibits growth of esophageal squamous cell carcinoma in vitro and patient-derived xenografts 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 4299.
Abstract 4302: AR signaling promotes prostate carcinogenesis
Authors: Cheng Siyuan , Yang Shu , Connelly Zachary M. , Chen Fenghua , Yu Xiuping
Published: August 2019

Prostate cancer (PCa) is the most common non-skin cancer in American men. Each year, about 164,690 new patients will be diagnosed and about 29,430 men will die from this disease in 2018. Studies have identified several factors that increase the risk of PCa, including age, race, family history, and some diet factors. However, the exact cause of PCa remains unclear. Prostate is an androgen dependent organ and the growth of PCa also dependents on androgen and AR signaling. It is well established that AR signaling is the central player in promoting PCa progression. Here, we explore if AR signaling also plays a role in prostate carcinogenesis. We found that activation of AR signaling in normal prostate epithelial cells suppressed their cell proliferation in vitro but induced carcinogenesis in vivo. To explore the mechanisms that contribute to the differential effects of AR in vitro and in vivo, we examined if AR signaling induces genome instability in prostate epithelial cells, resulting in carcinogenesis and tumor progression. Senescence-associated beta-galactosidase staining results indicated that androgen treatment induced cellular senescence of prostate epithelial cells. Also, we found that the cells that survived the cellular senescence had altered genome ploidy and increased expression of active DNA-PK, suggesting that non-homologous end joining repair pathway is active in these cells. Our data support that AR signaling promotes genome instability in prostate epithelial cells, contributing to carcinogenesis. This may explain the high frequency of cancer developed in hormone-regulated organs such as prostate, mammary gland, and liver.

Funding: This study was supported by: FWCC/Foundation Legacy Funds, DOD New Investigator Award, The Office of Research SEED awards, The Department of Biochemistry & Molecular Biology.

Citation Format: Siyuan Cheng, Shu Yang, Zachary M. Connelly, Fenghua Chen, Xiuping Yu. AR signaling promotes prostate carcinogenesis [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 4302.
Abstract 431: Semi-automated detection, isolation and molecular analysis of single disseminated melanoma cells from lymph nodes
Authors: Alberter Barbara , Scheitler Sebastian , Feliciello Giancarlo , Ferrarini Alberto , Werner-Klein Melanie , Haferkamp Sebastian , Klein Christoph A. , Polzer Bernhard
Published: August 2019

We recently showed that lymph node disaggregation followed by immunocytology enables precise quantification of disseminated cancer cells (DCCs) and demonstrated that this approach has a 20-fold higher sensitivity to detect melanoma DCCs than routine histopathology (Ulmer et al., PLoS Medicine, 2014). Moreover, genetic profiling of single melanoma DCCs identified a colonization signature consisting of specific copy number alterations and point mutations that identify patients with high risk of progression (Werner-Klein et al., Nature Communications 2018). Here, we present the adaptation of this method to a semi-automated workflow for detection, isolation and molecular analysis of single melanoma DCCs. The developed workflow includes a mechanical disaggregation of lymph node tissue and collection of the mononuclear cells, immunofluorescence staining against melanoma-associated markers gp100 and MCSP and depletion of CD45-positive cells. Individual melanoma cells are then detected and isolated by DEPArrayTM technology enabling single cell whole genome amplification (Ampli1TM) for subsequent molecular analysis. In total, we processed 20 lymph nodes of melanoma patients and detected melanoma DCCs in 11/20 samples (55%). The quality of isolated cells was checked by Ampli1TM QC and 174 isolated single cells were further analyzed by Sanger sequencing for specific point mutations (BRAF, NRAS and cKIT) and Ampli1TM low pass kit for Illumina for copy number variation (CNV). Successful molecular analysis was correlated with genome integrity score (GII) as determined by Ampli1TM QC, with more than 95% of cells with GII 3-4 showing good performance in low pass sequencing. In conclusion, a new DepArrayTM based application for marker-dependent single cell isolation from malignant melanoma lymph nodes was successfully established and tested on a cohort of 20 melanoma patients. Molecular analyses of isolated single cells confirmed the tumor origin by CNV profiling and mutational analysis of melanoma-associated mutations. In the future, this approach could help to select individualized therapies for melanoma patients.

Citation Format: Barbara Alberter, Sebastian Scheitler, Giancarlo Feliciello, Alberto Ferrarini, Melanie Werner-Klein, Sebastian Haferkamp, Christoph A. Klein, Bernhard Polzer. Semi-automated detection, isolation and molecular analysis of single disseminated melanoma cells from lymph nodes [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 431.
Abstract 4311: Mechanisms responsible for the inflammatory polarization of monocytes by subclinical low-dose LPS
Authors: Pradhan Kisha
Published: August 2019

Dynamic polarizations of innate leukocytes such as monocytes and macrophages have been increasingly recognized to play key roles in the pathogenesis and/or treatment of human disease. However, mechanisms responsible for distinct polarizations of monocytes are not well understood. Our previous studies indicate that monocytes can be persistently polarized into a low-grade inflammatory state by subclinical super-low dose LPS (bacterial lipopolysaccharide). In this current study, we further characterized the unique polarization of monocytes by super-low dose LPS and its underlying mechanisms. We observed that super-low dose LPS preferentially induced the expression of inflammatory mediators such as MCP-1 while potently suppressed the expression of homeostatic mediators such as Catalase and FPN. Super-low dose LPS activated the inflammatory signaling molecules such as p38, CamKII, and NFkB. In contrast, super-low dose LPS suppressed the basal activation of immune suppressing molecules such as AMPK and SIRT3. Taken together, our data reveal novel insights regarding the persistent polarization of inflammatory monocyte, an emerging key player in various human diseases including cancer.

Citation Format: Kisha Pradhan. Mechanisms responsible for the inflammatory polarization of monocytes by subclinical low-dose LPS [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 4311.
Abstract 4328: Epigenetic modulation of SSTR2 expression provides the potential to broaden PEN-221 treatment population
Authors: Perino Samantha , Quinn James , Freda Jessica , White Brian , Bailey Scott , Bhatia Viren , Sweryda-Krawiec Beata , Bilodeau Mark , Bloss Jeffrey D. , Whalen Kerry , Wooster Richard
Published: August 2019

The expression of somatostatin receptor subtype 2 (SSTR2) is increased in up to 80% of neuroendocrine tumors (NETs), 20-40% of small cell lung cancers (SCLC) and a number of other tumor types. As a cell surface receptor, SSTR2 is a logical target for a drug conjugate. However, marginal, or loss of, expression of SSTR2 in tumor tissues has potential to limit the benefit from such conjugates.

Recent published preclinical studies have demonstrated that epigenetic modulators, such as HDAC inhibitors, can result in increased SSTR2 expression with treatment. Given that the expression of SSTR2 can be variable from patient to patient and in some cases low to absent, we hypothesized that the combination of epigenetic modulators with an SSTR2 target therapy may provide a benefit for patients who are not eligible for these targeted approaches on their own.

PEN-221 is a miniature drug conjugate of an SSTR2 targeting peptide attached to the potent cytotoxic DM1 through a cleavable disulfide linker. PEN-221, currently in Phase 1/2a (NCT02936323), is designed as a potent and selective anticancer agent to treat patients whose tumors express SSTR2. The targeting peptide of PEN-221 selectively binds to SSTR2, triggers receptor internalization leading to the accumulation of the DM1 to provide antitumor activity by disrupting microtubule networks causing apoptosis and mitotic catastrophe. In preclinical studies, single agent PEN-221 treatment leads to complete and sustained tumor regression in xenograft models that over-express SSTR2. We hypothesize that increased expression of SSTR2 as a result of epigenetic modulation from HDAC or other epigenetic modulators, may allow for the broadening of PEN-221 treatable tumors.

To evaluate these hypotheses, we have combined PEN-221 and epigenetic modulators in preclinical models of cancer. Efficacy studies were carried out in models expressing various degrees of SSTR2, and combinations resulted in greater efficacy than that of the single agent. In addition, a pharmacodynamic assessment of epigenetic modulator treatment on SSTR2 expression levels was performed. These data demonstrate that combination of PEN-221 and epigenetic modulators provide greater efficacy than single agent activity alone and support the evaluation of such combinations in the clinical setting.

Citation Format: Samantha Perino, James Quinn, Jessica Freda, Brian White, Scott Bailey, Viren Bhatia, Beata Sweryda-Krawiec, Mark Bilodeau, Jeffrey D. Bloss, Kerry Whalen, Richard Wooster. Epigenetic modulation of SSTR2 expression provides the potential to broaden PEN-221 treatment population [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 4328.
Abstract 4332: Targeting MYC overexpressing leukemia with cardiac glycoside proscillaridin through downregulation of histone acetyltransferases
Authors: Costa Elodie Marie Da , Armaos Gregory , McInnes Gabrielle , Beaudry Annie , Moquin-Beaudry Gael , Bertrand-Lehouillier Virginie , Caron Maxime , St-Onge Pascal , Jonhson Jeffrey R. , Krogan Nevan , Sai Yuka , Downey Michale , Rafei Moutih , Boileau Meaghan , Eppert Kolja , Florez-Diaz Ema , Haman Andre , Hoang Trang , Sinnett Daniel , Beausejour Christian , McGraw Serge , Raynal Noel J.
Published: August 2019

Targeting MYC oncogene remains a major therapeutic goal in anticancer therapies. Here, we demonstrate that proscillaridin, a cardiac glycoside approved for heart failure treatment, causing Na+/K+ pump inhibition, targets efficiently MYC overexpressing cancer cells. At clinically relevant doses, proscillaridin induced rapid downregulation of MYC protein level, and produced growth inhibition preferentially against MYC overexpressing leukemic cell lines including lymphoid and myeloid stem cell populations. Whole transcriptome analysis with RNA sequencing of acute lymphoblastic leukemia cells showed a downregulation of gene sets involved in MYC pathways and cell replication, and an upregulation of genes involved in hematopoietic differentiation induced by proscillaridin treatment. Gene expression changes were associated with an epigenetic remodeling of chromatin active marks. Proscillaridin induced a significant loss of lysine acetylation in histone H3 (at lysine 9, 14, 18 and 27). In addition, mass spectrometry analysis revealed a loss of lysine acetylation in non-histone proteins such as MYC itself, MYC target proteins, and a series of histone acetylation regulators. Global loss of acetylation correlated with the rapid downregulation of histone acetyltransferase proteins (such as CBP, P300, TIP60 and GCN5) involved in histone and MYC acetylation. Overall, these results strongly support the repurposing of proscillaridin in MYC overexpressing leukemia and suggest a novel strategy to target MYC by inducing the downregulation of histone acetyltransferases involved in its stability.

Citation Format: Elodie Marie Da Costa, Gregory Armaos, Gabrielle McInnes, Annie Beaudry, Gael Moquin-Beaudry, Virginie Bertrand-Lehouillier, Maxime Caron, Pascal St-Onge, Jeffrey R. Jonhson, Nevan Krogan, Yuka Sai, Michale Downey, Moutih Rafei, Meaghan Boileau, Kolja Eppert, Ema Florez-Diaz, Andre Haman, Trang Hoang, Daniel Sinnett, Christian Beausejour, Serge McGraw, Noel J. Raynal. Targeting MYC overexpressing leukemia with cardiac glycoside proscillaridin through downregulation of histone acetyltransferases [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 4332.
Abstract 4340: A novel fluoride (ChlA-F) transcriptionally upregulates miR494 via a HuR/JunB axis to inhibit cell invasion in human bladder cancers
Authors: Tian Zhongxian , Hua Xiaohui , Xu Jiheng , Zhu Junlan , li Jingxia , Huang Chuanshu
Published: August 2019

Bladder cancer (BC) is a commonly occurring cancer and characterized by a low 5-year survival rate particularly in patients with invasive BCs. Here we investigated the chemotherapeutic activity of ChlA-F, a novel C8 fluoride derivative of cheliesisin A (ChelA) with potent anti-neoplastic properties. ChlA-F treatment upregulated miR494 expression and significantly suppressed cell invasion in human BC cell lines. Mechanistically, ChlA-F-induced upregulation of miR494 expression was due to a HuR-mediated increase in JunB mRNA stabilization and protein expression, which led to an increase in miR494 transcription via direct binding to the miR494 promoter region. After ChlA-F treatment, upregulated miR494 was found to bind to the 3’ UTR region of c-Myc mRNA, resulting in decreased c-Myc mRNA stability and protein expression. This, in turn, led to decreased transcription of a c-Myc downstream-regulated gene, MMP-2 and ultimately, inhibition of BC invasion. Here we provide the first evidence showing that our newly synthesized compound, ChlA-F, has a profound inhibitory effect on human BC invasion and elucidate the mechanisms underlying this anti-neoplastic activity. Our data suggest that ChlA-F may be a promising therapeutic alternative for treatment of invasive and metastatic human BC in patients.

Citation Format: Zhongxian Tian, Xiaohui Hua, Jiheng Xu, Junlan Zhu, Jingxia li, Chuanshu Huang. A novel fluoride (ChlA-F) transcriptionally upregulates miR494 via a HuR/JunB axis to inhibit cell invasion in human bladder cancers [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 4340.
Abstract 4347: Differentially expressed genes and molecular pathways in an autochthonous mouse prostate cancer model
Authors: Verma Shiv , Shukla Sanjeev , Pandey Mitali , MacLennan Gregory T. , Gupta Sanjay
Published: August 2019

Prostate cancer remains a major public health problem and second leading cause of cancer-related deaths in men in the United States. Despite widespread screening efforts and advancement in therapeutic regimens, incidence of prostate cancer remains steady in the past two decades. Although serum prostate-specific antigen (PSA) has been used as a screening test for prostate cancer for over two decades, the practice remains controversial. As a screening test for prostate cancer, PSA lacks sensitivity and specificity as early detection marker. Furthermore, screening may lead to unnecessary biopsies, over-detection, and overtreatment. There is a need for specific and sensitive biomarker(s) which can differentiate between cancer and non-cancer utilized for screening and early detection. The present study aims to comprehend the molecular pathways of prostate cancer which is essential for early detection and treatment. Dorso-lateral prostate from 20 week transgenic adenocarcinoma of the mouse prostate (TRAMP) mice, which spontaneously develops prostate cancer and recapitulates human disease and age-matched non-transgenic littermates were utilized for microarray analysis. Network and pathway analyses were mapped using the Ingenuity Pathway Analysis (IPA), and the database for annotation, visualization and integrated discovery. In total, 136 differentially expressed genes, including 32 downregulated genes and 104 upregulated genes were identified in the dorso-lateral prostate of TRAMP mice, compared to non-transgenic. A subset of differentially expressed genes were validated by QRT-PCR. Among these genes, 36% genes were connected to the nucleic acid binding especially ribosomal proteins which play important roles in protein synthesis, and is the most enriched pathway in the development of prostate cancer. Moreover, the results also suggest deregulation of the metabolic process and an imbalance between extracellular matrix components and adherens junction proteins could induce cancer progression. Taken together, the present study evaluated the underlying pathways and its connection to human cancer, which may further help to assess the risk of prostate cancer detection and its progression to identify potential targets of adjuvant treatment.

Citation Format: Shiv Verma, Sanjeev Shukla, Mitali Pandey, Gregory T. MacLennan, Sanjay Gupta. Differentially expressed genes and molecular pathways in an autochthonous mouse prostate cancer model [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 4347.
Abstract 4348: DNA-based fusion detection using a pan-cancer tumor profiling 532-oncogene panel
Authors: Dilger Katharine , Sun Yongming , Lai Kevin , Chakravarty Ushati Das , Sponer Nicole , Giorda Kristina , Lau Patrick , Wang Yu
Published: August 2019

Large-scale cancer profiling using next generation sequencing (NGS) has become instrumental to the discovery and identification of new, targetable cancer alterations. A comprehensive set of 532 oncogene targets were combined to create the new xGen® Pan-Cancer Panel V2 for hybrid capture sequencing. This xGen panel covers 2.2 Mb of the human genome, and allows for the simultaneous detection of copy number variations (CNVs), insertions and deletions (indels), gene rearrangements, and microsatellite instability across a wide range of sample types, inputs, and quality. Using a new library prep workflow optimized for low quality samples and low input, panel performance was first evaluated with 30 ng of input DNA using libraries built from matched samples [formalin-fixed paraffin-embedded (FFPE) tumor gDNA, frozen adjacent normal tissue gDNA, and cell-free DNA (cfDNA)] from five lung cancer donors (n = 15). Sample quality ranged from a mean DIN of 4.4 ±1.1 to 8.3 ±0.9 for FFPE tumor gDNA and frozen normal gDNA, respectively. After subsampling to 200X mean target coverage, 96% of target bases had at least 40X coverage for all libraries. Comparative hierarchal clustering analysis was then used to identify lung cancer mutations shared in all tumor samples. NGS gDNA reference standards from Horizon Discovery (HD753, HD798, HD799, and HD803) with verified CNVs, single nucleotide variations (SNVs), amplifications, and fusions, and were used to evaluate detection rates at different library input masses down to 1 ng. cDNA libraries were also prepared from RNA extracted from FFPE 5-Fusion RNA Multiplex Reference Standards (HD796, HD783). We identified all possible gene fusion events in the positive control using the structural variant caller, LUMPY (https://github.com/arq5x/lumpy-sv). The xGen Pan-Cancer Panel V2 enables a cost-efficient and time-saving approach for the detection of multiple oncogene targets.

Citation Format: Katharine Dilger, Yongming Sun, Kevin Lai, Ushati Das Chakravarty, Nicole Sponer, Kristina Giorda, Patrick Lau, Yu Wang. DNA-based fusion detection using a pan-cancer tumor profiling 532-oncogene panel [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 4348.
Abstract 4353: Stroma drive metabolic reprogramming and induces stemness properties in pancreatic cancer cells
Authors: Kesh Kousik K. , Gupta Vineet K. , Sharma Nikita , Hadad Roey , Dudeja Vikas , Saluja Ashok , Banerjee Sulagna
Published: August 2019

Pancreatic Ductal Adenocarcinoma (PDAC) is one of the most devastating human malignancies with poor survival rate owing to chemoresistance and relapse. Small population of Tumor Initiating Cells (TIC) are considered major players driving these traits. Recent literature has demonstrated the importance of microenvironment “niche” in enrichment of these TIC populations. The desmoplastic stroma in pancreatic cancer comprising of stromal cells and immune cells are instrumental in pathogenesis and progression of PDAC. Among many cytokines, IL-6 a pro-inflammatory cytokine plays a major role in it. In this study, we show that stromal cell secreted IL6 altered metabolic reprogramming and increased cancer stemness in PDAC cells. PDAC cells cultured with pancreatic stellate cells condition media (PSC-CM) or recombinant IL6 and stem ness markers expression were investigated. Cytometry Bead Array (CBA) were used for cytokine profiling. Metabolic alteration along with TIC gene expression were measured using flow cytometry and Seahorse. PDAC cells when cultured with PSC-CM exhibited increase glucose uptake along with an enrichment of CD133+ population. An analysis of the conditioned media using CBA reveled that PSC-CM is enriched with cytokine IL-6 and TNF-α. Treatment of PDAC cells with recombinant IL-6 increased expression of self-renewal genes SOX2, OCT4, Nanog along with biochemical marker ALDH. In addition, TIC marker CD133 and invasion marker gene MMP7 expression also increased upon IL6 induction. Moreover, IL6 treatment increased glucose uptake and lactate secretion. Small molecule inhibitor Stattic (inhibitor of STAT3) inhibit glucose uptake and stemness gene expression.Our study shows that stromal IL6 altered metabolic reprogramming in cancer cells as well as increased cancer stemness and invasion. In future, the underlying mechanisms of IL-6 driven metabolic alteration and its impact on stemness and metastasis will be investigated.

Citation Format: Kousik K. Kesh, Vineet K. Gupta, Nikita Sharma, Roey Hadad, Vikas Dudeja, Ashok Saluja, Sulagna Banerjee. Stroma drive metabolic reprogramming and induces stemness properties in pancreatic cancer cells [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 4353.