bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5746: CRISPR9-mediated FGL2-KO in tumor cells impairs tumor progression in brain and triggers antitumor immune memory systematically via NF-κB dependent Batf3 expression in DCs
Authors: Yan Jun , Zhao Qingnan , Gabrusiewicz Konrad , Kong Ling-Yuan , Huo Longfei , Xia Xueqing , Wei Jun , Xu Jingda , Davis Eric , Heimberger Amy B. , Li Shulin
Published: August 2018

Brain cancer is the 10th leading cause of death in United States with increasing primary tumor morbidity and secondary brain metastasis morbidity. Glioblastoma (GBM) is most lethal brain tumor with less than 5% 5 years survival rate marked by significant resistance to majority of therapies targeting tumor cells or tumor microenvironment. Here we shown one approach to convert aggressive GBM cells to immunogenic tumor cells by knockout FGL2 in GBM cells to evoke efficient anti-tumor immunosurvilliance. FGL2 is an immunosuppressive molecule and its expression level is strongly correlated with glioma grade and poor diagnosis. FGL2 from GBM cells, but not from host cells, is essential for immune evasion dependent GBM progression using both preclinical GBM and brain metastatic tumor models. Knockout FGL2 in GBM cells completely abolish tumor progression of GBM models challenged with low or high number of tumor cells in immunocompetent mice and protect mice from tumor rechallenge in periphery. This eradication effect is impaired in immune deficient mice and CD8+ T cells depleted mice, but not in FGL2KO mice, showing the necessarily of immune system to eliminate FGL2KO tumor cells and the requirement of FGL2 in tumor cells to evade immunosurvilliance. To understand how the systemic CD8+ T cell-dependent anti-tumor response and memory was induced by FGL2KO GBM cells, we studied the impact of FGL2KO tumor cells on dendritic cells (DCs) which control immune response via its critical role in the generation of effective cytotoxic T lymphocyte (CTL) responses. Knockout FGL2 in GBM cells induced CD8a+/CD103+ DCs accumulation in brain in vivo; DCs proliferation and CD103+ DCs induction in vitro. In agreement with this discovery, knockout Batf3, a key transcription factor regulating CD8a+/CD103+ DCs, reverse the impact of FGL2KO in GBM cell in vivo. By molecule analysis, a novel signaling pathway was revealed that knockout FGL2 in GBM cells induce NF-κ B activation, and this activation induced Batf3 and associated CD103 expression in DCs. Therefore, target specific molecule in GBM cells, such as knockout FGL2, can convert aggressive GBM cells to immunogenic cells, pointing to new approach for immune therapy.

Citation Format: Jun Yan, Qingnan Zhao, Konrad Gabrusiewicz, Ling-Yuan Kong, Longfei Huo, Xueqing Xia, Jun Wei, Jingda Xu, Eric Davis, Amy B. Heimberger, Shulin Li. CRISPR9-mediated FGL2-KO in tumor cells impairs tumor progression in brain and triggers antitumor immune memory systematically via NF-κB dependent Batf3 expression in DCs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5746.
Abstract 5749: Interleukin-1 (IL-1) induces a treatment-resistant gene expression pattern in prostate cancer (PCa)
Authors: Thomas-Jardin Shayna Elizabeth , Kanchwala Mohammed , Jacob Joan , Meade Rachel , Gahnim Nagham , Delk Nikki
Published: August 2018

Inflammation promotes the development and progression of cancer, where inflammatory cytokines secreted by infiltrating immune cells and tumor cell autocrine signaling are usurped to promote tumor cell survival and disease progression. The interleukin-1 (IL-1) inflammatory cytokine is elevated in prostate cancer (PCa) patient tissue and serum, where it promotes PCa metastasis and bone colonization. Performing in vitro analysis on PCa cell lines and RNA sequencing, we found that IL-1 concomitantly upregulates many pro-survival and lineage proteins, and represses the hormone receptor, androgen receptor (AR). Despite AR loss, a subpopulation of PCa cells remain viable. Our observation could reflect a role for IL-1 in the de novo accumulation of AR- PCa subpopulations. Indeed, AR- PCa subpopulations, such as stem and neuroendocrine cells, are implicated in PCa resistance to AR-targeted therapies. Thus, we hypothesize that IL-1 may reprogram AR+ PCa cells into treatment-resistant AR- PCa cells that survive hormone receptor repression through the upregulation of pro-survival pathways. To begin to address this hypothesis, we performed RNA sequencing and pathway analysis to identify an IL-1-conferred expression pattern in the LNCaP AR+ PCa cell line that mimics the PC3 AR- PCa cell line. We identified canonical inflammatory and immune response pathways, including NFκB, IL-8, and iNOS signaling cascades, likely facilitating PCa cell survival in an inflammatory tumor microenvironment. We also identified novel IL-1-regulated proteins that are known to promote PCa cell survival, such as SQSTM1/p62, SOX9, and ELF3. Experiments are underway to determine the functional significance of our IL-1-conferred gene expression profile in cell line and PDX models. Our investigation will provide insight into the mechanistic function of IL-1 in PCa resistance to AR-targeted therapies

Citation Format: Shayna Elizabeth Thomas-Jardin, Mohammed Kanchwala, Joan Jacob, Rachel Meade, Nagham Gahnim, Nikki Delk. Interleukin-1 (IL-1) induces a treatment-resistant gene expression pattern in prostate cancer (PCa) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5749.
Abstract 5750: Underlying inflammatory processes in non-neoplastic female thyroid serve as an immunological basis for the sex disparity in papillary thyroid cancer
Authors: Gupta Anvita , Singh Karnika , O'Connell Timmy J. , Jones Melanie , Rasamny JK , Schwarcz Monica , Moscatello Augustine , Shin Edward , Tiwari Raj , Geliebter Jan
Published: August 2018

Background: The American Cancer Society (ACS) estimated 56,870 new cases of thyroid cancer in the United States in 2017, with roughly 2,010 deaths attributed to the disease. The incidence of thyroid cancer has been increasing steadily over the past three decades, with Papillary Thyroid Cancer (PTC) being the most prevalent form of this endocrine malignancy, comprising of about 90% of all cases. Appreciably, Surveillance, Epidemiology, and End Results (SEER) and the ACS indicate that the incidence of PTC in the age group of 20-49 years is three to four times more prevalent in women than in men. This epidemiologic finding suggests that the female thyroid is at a higher risk of developing PTC.

Methods: The Cancer Genome Atlas (TCGA) database provides the means to address the differences between male and female thyroids on the molecular level. Using the normalized Level 3 RNA Seq Data and patient pathology reports available on the TCGA Data Portal, differential gene expression of 56 non-neoplastic/normal thyroid samples (40 women; 16 men) was investigated using the biological analysis software, Subio platform, version v1.20.5031. (Subio Inc., Tokyo, Japan).

Results: Immune/inflammatory genes exhibited a higher expression in uninflamed, normal thyroid tissue of women than in men (>1.5 fold; p<0.05). The expression levels of these genes were increased in male PTC samples, compared to their matched normal tissue. The level of expression of immune/inflammatory genes in women approximated their expression levels in matched PTC tissue of women and men, indicating that normal thyroid tissue in women exhibits a neoplastic level of inflammation.

Conclusions: Our data suggest that the female thyroid may be “at-risk” for developing PTC due to underlying and on-going immune/inflammatory processes.

Citation Format: Anvita Gupta, Karnika Singh, Timmy J. O'Connell, Melanie Jones, JK Rasamny, Monica Schwarcz, Augustine Moscatello, Edward Shin, Raj Tiwari, Jan Geliebter. Underlying inflammatory processes in non-neoplastic female thyroid serve as an immunological basis for the sex disparity in papillary thyroid cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5750.
Abstract 5768: MG1122, a whole IgG-like bispecific antibody targeting mesothelin and CD3, induces T cell-mediated killing of MSLN-expressing tumor cells
Authors: Lee Yun-Jung , Lim Okjae , Kim Munkyung , Lee Jeewon , Kim Kisu , Jung Junhong , Lee SuA , Kim Sung Keun , Kwon Haenaem , Lim Yangmi , Park Yong Yea , Won Jonghwa
Published: August 2018

Mesothelin (MSLN) is an antigen overexpressed in several malignancies, including mesothelioma and ovarian and pancreatic adenocarcinoma. It has been studied as a marker for diagnosis and a target for immunotherapy. Here, we adopted a bispecific antibody format for recruiting cytotoxic T cells to kill tumor cells. MG1122 is a novel, whole IgG-like bispecific antibody which recognizes CD3 on T cells and MSLN on tumor cells. MG1122 induced effective killing of MSLN-expressing tumor cells. This response was associated with robust activation of T cells as shown by nuclear factor of activated T cells (NFAT) activation, CD25 and CD69 upregulation and increased cytokine release. In addition, using a live cell analysis system, we found that levels of activated caspase 3/7 were amplified in the tumor cells by MG1122 treatment. Although MSLN is a surface antigen, it also exists as a secreted isoform, which can lead to tumor evasion against MSLN targeting antibodies. However, despite the high concentration of soluble MSLN, MG1122 still effectively bound and showed strong killing effects against MSLN-expressing tumor cells. Taken together, our MSLN/CD3 bispecific antibody, MG1122 offers a promising opportunity to redirect T cells to kill MSLN-expressing cancer cells.

Citation Format: Yun-Jung Lee, Okjae Lim, Munkyung Kim, Jeewon Lee, Kisu Kim, Junhong Jung, SuA Lee, Sung Keun Kim, Haenaem Kwon, Yangmi Lim, Yong Yea Park, Jonghwa Won. MG1122, a whole IgG-like bispecific antibody targeting mesothelin and CD3, induces T cell-mediated killing of MSLN-expressing tumor cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5768.
Abstract 577: Oral microbiome biomarker panel to detect oral and oropharyngeal cancers in a clinical setting
Authors: Lim Yenkai , Fukuma Naoki , Totsika Makrina , Kenny Liz , Morrison Mark , Punyadeera Chamindie
Published: August 2018

Oral microbiome plays a role in the instigation of oral diseases that affect other systemic conditions. These associations encourage the exploration of oral dysbiosis leading to the pathogenesis of cancers. In this study, saliva was used to characterise the oral microbiome fluctuation associated with oral and oropharyngeal cancers. The study cohort consists of normal healthy controls (n = 10, between 20 to 30 years of age; n = 10, above 50 years of age), high-risk individuals (n = 11, above 50 years of age with bad oral hygiene and/or oral diseases) and oral and oropharyngeal cancer patients (n = 31, HPV-positive; n = 21, HPV-negative). Saliva samples were analysed using 16S rRNA gene amplicon sequencing on the MiSeq platform. Kruskal-Wallis rank test was used to identify genera associated with oral and oropharyngeal cancers. A logistic regression analysis was carried out to determine the performance of these genera as a biomarker panel to detect oral and oropharyngeal cancers. In addition, a two-fold cross-validation with a bootstrap procedure was carried out on R to investigate how well the panel would perform in a real clinical scenario. Our data indicates that the oral microbiome is able to predict the presence of oral and oropharyngeal cancers with sensitivity and specificity of 100% and 90% respectively. With further validation, the panel could potentially be implemented into clinical workflow as oral and oropharyngeal cancers diagnostic and prognostic biomonitors.

Citation Format: Yenkai Lim, Naoki Fukuma, Makrina Totsika, Liz Kenny, Mark Morrison, Chamindie Punyadeera. Oral microbiome biomarker panel to detect oral and oropharyngeal cancers in a clinical setting [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 577.
Abstract 5785: EML4-ALK variant E6a/b;A20 positive NSCLC cell lines are associated with growth upon blocking MEK-ERK pathway
Authors: Hallberg Bengt , Palmer Ruth H. , Umapathy Ganesh , Gustafsson Dan E. , Siaw Joachim T. , Alam Wasi , Guan Jikui , Doebele Robert , Le Anh T. , Doak Andrea
Published: August 2018

Background:

The protein kinase B (PKB/AKT) and RAF/MEK/ERK signaling pathways are activated in a wide range of human cancer types. In many cases, concomitant inhibition of both pathways is necessary to block proliferation and induce cell death and tumor shrinkage. Several feedback mechanisms have been described in which inhibition of one pathway leads to activation of a parallel signaling pathway, thereby decreasing the effectiveness of targeted monotherapies.

Materials and Methods:

In order to determine which signalling core should be blocked for combinatorial treatment, we treated a panel of EML4-ALK positive lung cancer cell lines using MEK-ERK inhibitors. Resazurin assay was performed to evaluate cell viability. Protein levels were determined using western blotting.

Results:

In this study, we describe a feedback mechanism in which MEK-ERK inhibition leads to increased activation of AKT signaling in EML4-ALK variant (E6a/b;A20) positive NSCLC cell lines. Interestingly, EML4-ALK variant (E13;A20) NSCLC cell lines responds to MEK-ERK inhibitors and shows synergism when combined with ALK tyrosine kinase inhibitors. We found that feedback response in EML4-ALK variant (E6a/b;A20) positive NSCLC cells was mediated by the mammalian target of rapamycin complex 2-associated protein SIN1, resulting in increased survival and proliferation that depended on AKT signaling.

Conclusions:

Taken together, these results elucidate an important feedback network and contraindicate the use of MEK inhibitors as effective therapeutic strategy in EML4-ALK variant (E6a/b;A20) positive NSCLC.

Citation Format: Bengt Hallberg, Ruth H. Palmer, Ganesh Umapathy, Dan E. Gustafsson, Joachim T. Siaw, Wasi Alam, Jikui Guan, Robert Doebele, Anh T. Le, Andrea Doak. EML4-ALK variant E6a/b;A20 positive NSCLC cell lines are associated with growth upon blocking MEK-ERK pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5785.
Abstract 5806: Combination of the miniature drug conjugate PEN-866 with PARP inhibitors as a rational approach to overcoming limitations of PARP inhibitor monotherapy
Authors: Perino Samantha , Quinn James M. , Callahan Marrissa , Sharma Gitanjali , White Brian , Bilodeau Mark , Alland Leila , Whalen Kerry , Wooster Richard
Published: August 2018

PEN-866 is a novel miniature drug conjugate comprised of a Heat Shock Protein 90 (HSP90) targeting ligand attached through a cleavable linker to SN-38, the active metabolite of the approved topoisomerase I inhibitor, irinotecan. The conjugate accumulates and is retained in tumor cells through the HSP90 targeting ligand. PEN-866 linker cleavage provides sustained release of SN-38 at a high local tumor concentration leading to DNA damage and apoptosis of tumor cells resulting in broad preclinical antitumor activity. Reducing the DNA repair activity in tumor cells is an opportunity to significantly potentiate the efficacy of PEN-866.

Poly ADP ribose polymerase (PARP) has an important role in the repair of DNA. PARP inhibitors have shown activity in the clinical setting, both in patients whose cancers harbor mutations in BRCA1 and BRCA2, and in those that are BRCA1/2 wildtype, and are approved as monotherapy for use in treatment of patients with advanced ovarian cancers. Combining PARP inhibitors with other therapeutics has been challenging due to dose limiting toxicities. Combining PARP inhibitors with PEN-866 is mechanistically appealing with the potential for greater efficacy as compared to these agents used alone and the potential to mitigate the toxicity of the combination of a PARP inhibitor with conventional cytotoxic therapy due to the high levels of accumulation and retention of PEN-866 in xenograft tumors.

To evaluate these hypotheses, we have combined PEN-866 and PARP inhibitors in preclinical models of human cancer. Efficacy studies were carried out in both BRCA mutant and BRCA wildtype tumor xenografts combining PEN-866 and PARP inhibitors, and the combinations resulted in greater efficacy than that of the single agent in tumors with both genetic backgrounds. In addition, a pharmacodynamic assessment of DNA damage was performed in tumors responsive to the combination treatment. These data demonstrate that combination of PEN-866 and PARP inhibitors provide greater efficacy than single agent activity alone, including BRCA wildtype tumors, and support the evaluation of such combinations in the clinical setting. The authors acknowledge David Proia, Weiwen Ying and others from Synta Pharmaceuticals who contributed to this work.

Citation Format: Samantha Perino, James M. Quinn, Marrissa Callahan, Gitanjali Sharma, Brian White, Mark Bilodeau, Leila Alland, Kerry Whalen, Richard Wooster. Combination of the miniature drug conjugate PEN-866 with PARP inhibitors as a rational approach to overcoming limitations of PARP inhibitor monotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5806.
Abstract 5814: Synthetic lethal therapy to overcome resistance to AURKA inhibition in HPV-negative head and neck cancer
Authors: Deneka Alexander Y. , Einarson Margret B. , Lee Jong Woo , Nikonova Anna S. , Burtness Barbara , Golemis Erica A.
Published: August 2018

Head and neck squamous cell carcinomas (HNSCC) affect more than a half million people annually worldwide. Most currently available treatments, which include cisplatin and irradiation, are associated with significant morbidity, while many targeted inhibitors show limited single agent efficacy: novel therapeutic approaches which combine efficacy and limited toxicity are needed. The most common mutational event in human papillomavirus (HPV)-negative HNSCC is mutation of TP53. As control of the G1/S transition is entirely p53-dependent, cells with TP53 mutations lose the G1/S checkpoint, and evade radiation- and cisplatin-induced cellular senescence. Targeting the serine/threonine mitotic kinase Aurora-A (AURKA) emerged as a promising synthetic lethal approach in this setting. AURKA is negatively regulated by p53, which causes upregulation of AURKA in the majority of cases of HNSCC, correlating with poor prognosis and cisplatin resistance. The single agent AURKA inhibitor MLN8237 (alisertib) had a 9% monotherapy response rate in treatment-refractory HNSCC, with all responses occurring in HPV-negative or -unknown disease. In our study, we used initial high throughput screening in multiple genomically characterized HNSCC cell lines to identify synergistic combinations involving AURKA inhibitors and 20 other inhibitors of cell cycle, and cell cycle checkpoint controls. This work has identified has several effective combinations, particularly in between alisertib (targeting AURKA), danusertib (AURKA and AURKB), LY2606368/prexasertib (CHK1), and reversine (MPS1). Multiplexing analysis with measures of cell metabolism, direct visualization of mitotic nuclei, and additional assays has allowed us to dissect distinct execution points for different drug combinations. We will describe in detail mechanistic interactions between specific combinations of value for HNSCC. These drug combinations are likely to represent a new and effective modality for treating TP53-mutated cancers.

Citation Format: Alexander Y. Deneka, Margret B. Einarson, Jong Woo Lee, Anna S. Nikonova, Barbara Burtness, Erica A. Golemis. Synthetic lethal therapy to overcome resistance to AURKA inhibition in HPV-negative head and neck cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5814.
Abstract 582: Accelerating drug development with a CLIA approved functional test using patient derived organoids
Authors: Schaub Franz X. , Churchill Michael J. , Swan Hallie A. , Rosati Rachele , Watt Roland M. , Shaw Reid C. , Murphy Stephanie A. , Diaz Robert L. , Pereira Shalini C. , Grandori Carla
Published: August 2018

The pool of targeted therapies is expanding steadily providing novel treatment opportunities. However, the process of expanding indications and including additional cancer types after initial approval is slow. Here we present an in vitro clinical trial platform using the data from the first CLIA-approved Precision Medicine Platform using high-throughput screening (P.A.R.I.S. Assay, SEngine Precision Medicine, Seattle, WA).

Briefly, patient-derived tumor cells from biopsies are expanded as living organoids, and functionally evaluated using a high-throughput drug screen. The drug library currently includes FDA approved and experimental drugs. Results are integrated with known genomic information and reported to the clinician to highlight treatment options of an individual patient. Combining this data from multiple patients enables the discovery of novel indications and biomarkers. To date we have acquired data from a wide range of tumor types including hard-to-treat cancers, rare tumor types and tumors from heavily pretreated patients. In addition, we also have collected data from more than 100 patient derived cell lines.

Using the data analysis platform developed in R, the drug responses across all patient screens of a given drug can be analyzed to find common indicators of sensitivities. For instance, a BRD4 inhibitor (CPI-203), which was tested on 8 different primary tumor samples, was selectively active in a subset of the patients. Interestingly, most of the responding tumor types are ovarian and breast cancer cases with mutations in BRCA1 or EZH2. However, there are currently no clinical trials investigating the more developed BRD4 inhibitor CPI-0610 for breast or ovarian cancer. Non-responders, on the other hand, are spread between multiple cancer types.

The number of patients receiving the P.A.R.I.S. test is steadily increasing and these patient cases contribute to our knowledge base. As this test is used to inform clinicians about potential treatment opportunities, we are also accumulating information on how these patients respond to treatment. Overall, this platform will provide real world data for optimal drug combinations and novel biomarkers to quickly expand indications of existing drugs.

Citation Format: Franz X. Schaub, Michael J. Churchill, Hallie A. Swan, Rachele Rosati, Roland M. Watt, Reid C. Shaw, Stephanie A. Murphy, Robert L. Diaz, Shalini C. Pereira, Carla Grandori. Accelerating drug development with a CLIA approved functional test using patient derived organoids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 582.
Abstract 5842: Oxidative stress and protein carbonylation towards multi-drug resistance in cancer
Authors: Aryal Baikuntha P. , Gillet Jean-Pierre , Rao V Ashutosh
Published: August 2018

Reactive oxygen species (ROS) are constantly produced under normal as well as stress conditions and play important roles in cell signaling and homeostasis. An elevated level of ROS has been reported in most cancers which may play a critical role in both cancer progression or cancer suppression. One proposed therapeutic strategy is to alter the oxidative stress from ROS-induced tumor promoting event to a ROS-induced apoptotic signaling. Most ROS inducing therapeutic agents initially induce ROS to kill cancer cells by apoptosis; however, prolonged exposure of drugs may result in reduced ROS level and an adaptive microenvironment that makes cancer cells resistant to the same ROS-inducing therapeutic drugs. Chronic exposure of ROS may also upregulate the multidrug resistance protein expression in cancer cells. We hypothesize that differences in oxidative stress and the antioxidant response between drug sensitive and drug resistance cells may play a key role in drug resistance to redox-active drugs. We used two pairs of cell lines, a mitoxantrone sensitive and resistance MCF7 human breast adenocarcinoma (MCF7/MCF7MX) and H460 non-small lung cancer cell lines (H460/H460MX) for our experiments. We employed western blot and ELISA assays to determine the total and specific protein oxidation by carbonylation. Both MCF7MX and H460MX showed slightly lower level of total protein carbonylation compared to their mitoxantrone sensitive counterparts. In western blot analysis, we observed two specific proteins that were differentially carbonylated in MCF7MX compared to and MCF7. Both cytoskeletal and multifunctional proteins were identified by mass spectrometry and confirmed by two-color western blot analysis. Investigation of the potential roles of these specific protein carbonylation in drug resistance will be presented.

Citation Format: Baikuntha P. Aryal, Jean-Pierre Gillet, V Ashutosh Rao. Oxidative stress and protein carbonylation towards multi-drug resistance in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5842.
Abstract 5865: Gartanin, a 4-prenylated xanthone in the mangosteen fruit, inhibits the growth of prostate cancer cells associated with Skp2 degradation via neddylation inhibition and FBXW2 induction
Authors: Pham Victor , Bui Michelle , Rendon Raymond , Agredano Ericka , Le Thanh , Song Liankun , Zi Xiaolin
Published: August 2018

The S-phase kinase-associated protein 2 (Skp2) is overexpressed in prostate cancer and associated with tumor stage, disease recurrence, and worse patient survival. Studies also have demonstrated that Skp2 overexpression plays a critical role in development and progression of prostate cancer in several prostate cancer models, including PTEN knockout and TRAMP carcinogenesis and CWR22 xenograft models. Therefore, Skp2 is a promising target for developing new therapeutic and preventive approaches for prostate cancer. Here, we have found that gartanin, 4-prenylated xanthone from the Garcinia mangostana fruit (mangosteen), promoted Skp2 degradation in a proteasome-dependent manner in prostate cancer cells. In addition, gartanin induced the expression of FBXW2, a novel E3 ligase of SKP2 for targeted degradation, and downregulated the expression of NEDD8, leading to reduced neddylation levels of UBC12 and Cullin1. Molecular modelling analysis predicts that gartanin reasonably docks onto the regulatory subunit of NEDD8 activating enzyme E1 (NAE1) and next to the NEDD8 binding complex. More interestingly, gartanin selectively inhibits the growth of Skp2 overexpressing PC3 cells compared to parental PC cells (IC50 values for PC3/Skp2 vs. PC3 cells are approximately 4 μM vs. 14 μM). Together, our data indicate that gartanin is a novel natural agent for targeting Skp2 degradation via induction of FBXW2 expression and inhibition of neddylation. Gartanin deserves further investigation for its usefulness in prostate cancer prevention and treatment.

Citation Format: Victor Pham, Michelle Bui, Raymond Rendon, Ericka Agredano, Thanh Le, Liankun Song, Xiaolin Zi. Gartanin, a 4-prenylated xanthone in the mangosteen fruit, inhibits the growth of prostate cancer cells associated with Skp2 degradation via neddylation inhibition and FBXW2 induction [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5865.
Abstract 5878: Effect of cannabinoid WIN 55,212-2 on prostate cancer cell proliferation, migration, invasion, and tumor growth
Authors: Roberto Domenica , Klotz Laurence H. , Venkateswaran Vasundara
Published: August 2018

Introduction and Objective: Prostate cancer (PCa) is the most commonly diagnosed cancer in men and the second leading cause of cancer related death in Canada. A large body of evidence supports a possible role for cannabinoids in certain aspects of human health and disease, acting as palliative agents and more importantly, as inhibitors of cancer cell proliferation, migration, invasion, and the angiogenesis of tumours. WIN 55,212-2 (WIN) is a highly potent synthetic cannabinoid, binding to the cannabinoid receptors 1 and 2. This study aims to determine the anti-cancer effect of WIN using preclinical models of prostate cancer.

Methodology: Human PCa cells (DU145, PC3) were treated with WIN at concentrations ranging from 1μM to 20μM and growth of cells (using the MTS assay) was assessed at various times following treatment. Wound-healing assays were conducted to investigate the migratory potential of cells following exposure to treatment, and trans-well invasion assays were performed to explore the influence of WIN on cell invasion. In vivo evaluation using PC3 xenografts was performed using swiss athymic mice. Treatment was administered thrice weekly and tumor volume assessed for a total of three weeks.

Results: Treatment with 10μM WIN resulted in a significant reduction in the proliferation of DU145 and PC3 cells after 24 h compared to control (p<0.05). Cell migration and invasion studies revealed a significant reduction in cell motility at 15μM WIN (p<0.05) and a significant reduction in cell invasion at a concentration as low as 1μM (p<0.05). Treatment with 5mg/kg WIN (ip) revealed significant differences in tumor volume compared to controls that received the vehicle (p<0.05). There were no significant alterations in body weight in both groups.

Conclusion: WIN 55,212-2 is a highly potent cannabinoid, having significant influences on cell proliferation, migration, and invasion, with reduction in the growth of tumors in prostate cancer xenografts. We are the first to demonstrate the use of WIN in PC3 xenografts and provide evidence for its use as a novel therapeutic option in patients with prostate cancer.

Funding/Conflicts of Interest: None

Citation Format: Domenica Roberto, Laurence H. Klotz, Vasundara Venkateswaran. Effect of cannabinoid WIN 55,212-2 on prostate cancer cell proliferation, migration, invasion, and tumor growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5878.
Abstract 5906: EGFR and SOX2 crosstalk determines EGFR-TKI
Authors: Lee An-Chun , Yu Chia-Cherng , Wang Yuan-Hung , Chou Yu-Ting
Published: August 2018

Activating mutations in the epidermal growth factor receptor (EGFR), while initiating lung tumorigenesis, render tumors susceptible to EGFR-tyrosine kinase inhibitors (TKIs). SOX2 functions as a cell fate determination factor in lung progenitor cells. However, how SOX2 expression affects TKI treatment and tumor invasion in lung cancer is obscure. Here we report that EGFR mutations crosstalk with SOX2 to maintain cell proliferation and barrier properties in lung cancer cells, some of which generate heterogeneity by switching off SOX2 expression, thus promoting TKI resistance and invasiveness. Downregulation of SOX2 expression by shRNA against SOX2 or TGF-beta treatment induced epithelial-to-mesenchymal transition (EMT) and endowed cell resistant to TKIs. TKI treatment selected EGFR-mutated lung cancer cells harboring low SOX2 expression with the EMT signature. TKI naïve EGFR-mutated lung cancer cells harboring low SOX2 expression exhibits intrinsic TKI resistance. Conversely, enhancing endogenous SOX2 expression reverses EMT and increases sensitivity to TKIs. Immunohistochemistry analysis revealed that low SOX2 expression predicts a poor survival in patients harboring EGFR mutations. Taken together, our findings show how cancer plasticity elicited by SOX2 expression and TKI selection generates distinct oncogenic properties and TKI sensitivity, providing critical insights into tumor progression in EGFR-mutated lung cancer.

Citation Format: An-Chun Lee, Chia-Cherng Yu, Yuan-Hung Wang, Yu-Ting Chou. EGFR and SOX2 crosstalk determines EGFR-TKI [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5906.
Abstract 595: Perioperative risk factors for postoperative pneumonia after major oral cancer surgery: A retrospective analysis of 331 cases
Authors: Xu Jieyun , Yu Pei , Wang Weiwang , Zhuang Zehang , Liu Xiqiang
Published: August 2018

Objective: Postoperative pneumonia (POP) is common and results in prolonged hospital stays, higher costs, increased morbidity and mortality. However, data on the incidence and risk factors of POP after oral and maxillofacial surgery are rare. This study aims to identify perioperative risk factors for POP after major oral cancer (OC) surgery.

Methods: Perioperative data and patient records of 331 consecutive subjects were analyzed in the period of April 2014 to March 2016. We individually traced each OC patient for a period to discharge from the hospital or 45 days after surgery, whichever occur later.

Results: The incidence of POP after major OC surgery with free flap construction or major OC surgery was 11.6% or 4.5%, respectively. Patient-related risk factors for POP were male sex, T stage, N stage, clinical stage and preoperative serum albumin level. Among the investigated procedure-related variables, incision grade, mandibulectomy, free flap reconstruction, tracheotomy, intraoperative blood loss, and the length of the operation were shown to be associated with the development of POP. Postoperative hospital stay was also significantly related to increased incidence of POP. Using a multivariable logistic regression model, we identified male sex, preoperative serum albumin level, operation time and postoperative hospital stay as independent risk factors for POP.

Conclusion: Several perioperative risk factors can be identified that are associated with POP. At-risk oral cancer patients should be subjected to intensified postoperative pulmonary care.

Citation Format: Jieyun Xu, Pei Yu, Weiwang Wang, Zehang Zhuang, Xiqiang Liu. Perioperative risk factors for postoperative pneumonia after major oral cancer surgery: A retrospective analysis of 331 cases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 595.
Abstract 625: Cytotoxic T lymphocyte antigen 4 (CTLA-4) expression in ductal carcinoma in situ (DCIS) of the breast
Authors: Novo Jorge E. , Rosen Lauren , Ubago Julianne M. , Pincus Jennifer , Blanco Luis , Siziopikou Kalliopi P.
Published: August 2018

Background: Ductal carcinoma in situ (DCIS) consists of a heterogeneous group that can be subclassified by molecular signatures. High grade DCIS is commonly associated with periductal inflammation, a feature associated with increased risk for microinvasion. We previously assessed the immune microenvironment of DCIS and reported that HER2+ DCIS has increased numbers of tumor infiltrating lymphocytes (TILs), with significant expression of PD-L1. CTLA-4 is a supplementary immunomodulatory marker, which serves as a negative regulator of T-cell response. Targeted therapy is currently available against both PD-L1 and CTLA-4, and studies suggest an enhanced response with combination therapy as these therapies act though different mechanisms. In this study we evaluated the expression of CTLA-4 in both TILs and tumor cells in DCIS.

Design: The study population consisted of 74 cases of DCIS treated with surgical excision from 2008-2012. The molecular subtypes of DCIS were determined based on ER and HER2 expression. Tissue microarrays were constructed (3 cores/case) to account for tumor heterogeneity. The TMAs were immunostained for CTLA-4 and the expression CTLA-4 was assessed.

Results: Of the 74 cases, 41 were classified as Luminal A (ER+/HER2-), 17 as Luminal B (4 as Luminal B HER2- [ER+/HER2-/high Ki-67] and 13 as Luminal B HER2+ [ER+/HER2+]), 11 as HER2+ (ER-/HER2+) and 5 as basal (ER-/HER2-). Overall, the DCIS tumor cells expressed CTLA-4 in 58% of Luminal B (HER2+ 53% [7/13]; HER2- 75% [3/4]) and 36% of HER2+ (4/11), while only 39% of Luminal A (16/41), and one basal DCIS (1/5) did. In addition, the presence of CTLA-4 positive TILs was strongly associated with HER2+ expression (Luminal B HER2+=21.8%, HER2+=13.4%), compared to HER2- DCIS (Luminal B HER2-=8.5%, Luminal A=7.3%, Basal=6.4%) (p=0.026).

Conclusion: 1. CTLA-4 expression in TILs is increased in HER2+ DCIS. 2. A significant percent of tumor cells express CTLA-4 in all DCIS subtypes. Our results further advance our understanding of the role of the immune microenvironment in DCIS. Additional studies are needed to further elucidate the complex interplay between the host immune system and DCIS, which may offer insights into breast carcinoma progression and possible use of novel immune-based therapies.

Citation Format: Jorge E. Novo, Lauren Rosen, Julianne M. Ubago, Jennifer Pincus, Luis Blanco, Kalliopi P. Siziopikou. Cytotoxic T lymphocyte antigen 4 (CTLA-4) expression in ductal carcinoma in situ (DCIS) of the breast [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 625.
Abstract 637: Quantitative screening of serum protein biomarkers with reverse phase protein arrays
Authors: Kuang Zhizhou , Huang Ruochun , Lv Zhiqiang , Lv Jingqiao , Wu Jian , Huang Ruo-Pan
Published: August 2018

Screening biomarkers in serum samples for different diseases has always been of great interest because it presents an early, reliable, and, most importantly, noninvasive means of diagnosis and prognosis. Reverse phase protein arrays (RPPAs) are a high-throughput platform that can measure single or limited sets of proteins from thousands of patients' samples in parallel. They have been widely used for detection of signaling molecules involved in diseases, especially cancers, and related regulation pathways in cell lysates. However, this approach has been difficult to adapt to serum samples. Previously, we developed a sensitive method called the enhanced protein array to quantitatively measure serum protein levels from large numbers of patient samples. Here, we further refine the technology on several fronts: 1. simplifying the experimental procedure; 2. optimizing multiple parameters to make the assay more robust, including the support matrix, signal reporting method, background control, and antibody validation; and 3. establishing a method for more accurate quantification. Using this technology, we quantitatively measured the expression levels of 6 proteins: AFP, GP73, GDF15, IGFBP2, B2M and OPN from serum samples of 132 hepatocellular carcinoma (HCC) patients and 78 healthy volunteers. We found that all 6 protein expression levels are significantly increased in HCC patients. Statistical and bioinformatical analysis has revealed decent accuracy rates of individual proteins, ranging from 0.617 (B2M) to 0.908 (AFP) as diagnostic biomarkers to distinguish HCC from healthy controls. The combination of these 6 proteins as a specific HCC signature yielded a higher accuracy of 0.923 using LR, LDA, RF and SVM predictive model analyses. Our work reveals promise for using reverse phase protein arrays for biomarker discovery and validation in serum samples.

Citation Format: Zhizhou Kuang, Ruochun Huang, Zhiqiang Lv, Jingqiao Lv, Jian Wu, Ruo-Pan Huang. Quantitative screening of serum protein biomarkers with reverse phase protein arrays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 637.
Abstract 642: Comparison between in situ hybridization and digital polymerase chain reaction methods for detecting c-MYC gene copy number gain in tissue and cell-free plasma samples of colorectal cancer patients
Authors: Seo Soo Hyun , Lee Kyu Sang , Nam Soo Kyung , Park Kyoung Un , Oh Heung-Kwon , Kim Duck-Woo , Kang Sung-Bum , Lee Hye Seung
Published: August 2018

Introduction: We focused on the utility of the droplet digital polymerase chain reaction (ddPCR) for detecting c-MYC gene copy number (GCN) gain in cell-free plasma and tumor tissue of CRC patients.

Methods: c-MYC GCN was determined by using dual-color silver in situ hybridization (SISH) and ddPCR in retrospective cohort of 193 CRC patients (cohort 1) and prospective cohort of 64 CRC patients (cohort 2). Additionally, c-MYC GCN was analyzed by ddPCR method in cell-free plasma samples of cohort 2.

Results: In cohort 1, c-MYC GCN gain, defined as mean c-MYC copies/nucleus ≥ 4.0 in SISH analysis, was observed in 34 (17.5%), and c-MYC GCN gain by ddPCR method in 7 (3.6%). c-MYC GCN by SISH was significantly correlated with ddPCR results (ρ= 0.532, P < 0.001). Forty cases (20.7%) showed intratumoral genetic heterogeneity, but it did not affect the concordance between SISH and ddPCR results (P > 0.05). c-MYC GCN gain by both SISH and ddPCR methods had the worst prognosis (P = 0.001). In cohort 2, c-MYC GCN by SISH was significantly associated with tissue ddPCR (ρ=0.349, P = 0.005), but not with plasma ddPCR (P = 0.620). Intratumoral genetic and regional heterogeneity of c-MYC status have been found in 29 (45.3%) and 8 (12.7%). c-MYC status in plasma was associated with tissue ddPCR, but borderline statistical significance (ρ= 0.246, P = 0.050). Conclusions: Our findings suggested that results by SISH and ddPCR methods were discordant. Therefore, detecting method should be carefully selected to determine c-MYC GCN status.

Citation Format: Soo Hyun Seo, Kyu Sang Lee, Soo Kyung Nam, Kyoung Un Park, Heung-Kwon Oh, Duck-Woo Kim, Sung-Bum Kang, Hye Seung Lee. Comparison between in situ hybridization and digital polymerase chain reaction methods for detecting c-MYC gene copy number gain in tissue and cell-free plasma samples of colorectal cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 642.
Abstract 647: Identification of novel and known hotspot variants in African Americans with colorectal cancer
Authors: Brim Hassan , Tavakoli Payaam , Daremipouran Mohammad Reza , Shokrani Babak , Lee Edward , Nouraie Mehdi , Laiyemo Adeinko O. , Ashktorab Hassan
Published: August 2018

Background: The recent introduction of the cancer hotspot panel can help enhance targeted cancer therapy, but requires further refinement to identify which genes have significant clinical value for the population of interest. We aim to determine such genes by identifying somatic alterations in the 48 cancer gene panel in African Americans.

Methods: Tumor matched normal tissues (n=16) from African American colorectal cancer patients were analyzed using a 48 hotspot cancer panel by Ion Torrent sequencing. Variants of the genes found in TruSeq Amplicon - Cancer Panel (TSACP) were identified and validated. The type of variant was determined using bioinformatic pipelines including Ensembl's Variant Effect Predictor. The genes with relevant clinical value were determined based on frequency of somatic variants and results from previous studies.

Results: Somatic variants were detected in 39/48 (82%) genes. Of the 39 genes, the 7 with the most clinical value were selected for further analysis. Overall, 46 novel and known hotspot somatic variants were determined in the following 7 genes: KRAS (7/46 [15%]), NRAS (2/46 [4%]), MLH1 (1/46 [2%]), PIK3CA (8/46 [18%]), SMAD4 (6/46 [13%]), RET (9/46 [20%]), and TP53 (13/46 [28%]). Of the KRAS variants, 57% (4/7) were found in codons 12 and 13 while the other 43% (3/7) were found in codons 117 and 146. Out of the 46 somatic variants, 30% (14/46) were in hotspot regions. Of the KRAS hotspot variants, 100% (4/4) were in codons 12 and 13. The 2 NRAS variants were in codons 59 and 61. Lastly, the 9 genes without variants in our population are CDKN2A, SMARCB1, AKT1, ALK, FGFR2, IDH2, CSF1R, HNF1A and MPL.

Conclusion: We have identified novel and known hotspot somatic variants in African Americans with colorectal lesions. Patients with significant hotspot variants, primarily missense alterations, had a worse prognosis. As a result, cancer gene panel hotspot profile should be taken into consideration when treating African American CRC patients.

Citation Format: Hassan Brim, Payaam Tavakoli, Mohammad Reza Daremipouran, Babak Shokrani, Edward Lee, Mehdi Nouraie, Adeinko O. Laiyemo, Hassan Ashktorab. Identification of novel and known hotspot variants in African Americans with colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 647.
Abstract 668: Electron paramagnetic resonance (EPR) pO2 image-guided tumor biopsies to analyze hypoxia-induced proteins
Authors: Miller Richard , Epel Boris , Elas Martyna , Krzykawska-Serda Martyna , Maggio Matthew , Barth Eugene , Giurcanu Mihai , Halpern Howard
Published: August 2018

Stereotactic MCa4 and FSa tumor tissue biopsies registered with electron paramagnetic resonance (EPR) O2 imaging were used to identify sampling sites suitable to measure protein levels of three hypoxic cell biomarkers, hypoxia-induced factor 1-alpha (HIF-1α), carbonic anhydrase IX (CA9), and vascular endothelial growth factor (VEGF). EPR imaging provided quantitative localization of pO2 levels in tumors grown in the legs of C3H mice. Since hypoxic cells (pO2 <10 torr) are up to three times more resistant than normoxic cells to radiation lethality, identification of hypoxic populations to predict radiotherapeutic outcome is important. The oxygen broadening of narrow EPR spectral lines, or, equivalently, the increase in relaxation rates of electron magnetization, displays pO2 with 1-2 torr resolution in image voxels less than 1 mm3. The pO2 reporter molecule OX063 was used to image hypoxic areas in tumors. The molecule is selectively retained in tumors with a half-life of ~30 minutes. We used inversion recovery electron spin echo (IRESE) to measure the T1 rate of the trityl inside the tumor-bearing leg of mice. Voxel maps with O2 levels less than 10 torr (HF10) were used to guide biopsy cannulas to obtain tumor cell samples. Each of these biopsy cores were further subdivided into samples from which protein amounts could be determined via ELISA. Results from MCa4 and FSa tumor cells indicate a strong correlation between EPR pO2-identified hypoxic areas (<10 torr) and HIF-1α, CA9, and VEGF protein. The activation of various genes, including stem-cell activation, angiogenesis, vasodilation, glucose metabolism, reduced apoptotic activity, and cell cycle changes, contributes to poor radiotherapeutic outcome. EPR pO2 imaging may be used to precisely identify hypoxic areas harboring cells that protect tumor tissue from the toxicity of conventional radiation leading to radiotherapeutic failure.

Grant Support: This work was supported by grants from the NIH, including R01 CA98575 and P41 EB002034.

Citation Format: Richard Miller, Boris Epel, Martyna Elas, Martyna Krzykawska-Serda, Matthew Maggio, Eugene Barth, Mihai Giurcanu, Howard Halpern. Electron paramagnetic resonance (EPR) pO2 image-guided tumor biopsies to analyze hypoxia-induced proteins [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 668.