bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract LB-273: Identity card of tumor-infiltrating regulatory T cells in the context of tertiary lymphoid structures in lung cancer patients
Authors: DEVI Priyanka , Leveugle Sylvain , Goc Jeremy , Kaplon Helene , Germain Claire , Knockaert Samantha , Validire Pierre , Damotte Diane , Katsahian Sandrine , Fridman Wolf Herve , Sautes-Fridman Catherine , Lawand Myriam , Dieu-Nosjean Marie-Caroline
Published: July 2016

Regulatory T cells (Tregs) exert various suppressive mechanisms to dampen the host immune response which can help tumor cells to escape immune surveillance. However, the prognostic value of tumor-infiltrating Tregs (Ti-Tregs) is controversial. We described, tumor-associated tertiary lymphoid structures (TLS) in lung cancers, shown that they are critical for the local coordination and polarization of protective immunity and are able to imprint the behavior of intra-tumoral CD8+ T cells. The aim of the study was to determine the role of Ti-Tregs in the shaping of the local immune response in the context of TLS. We studied the differentiation, activation, immunosuppression, and immune checkpoint (ICP) status of Ti-Tregs in different areas of human lung tumors on a prospective cohort of 50 lung cancer specimens.

In lung cancer patients, CD3+FoxP3+ cells were detected in different zones of the tumor i.e. the tumor stroma (TLS and non-TLS areas) and to a lesser extend in tumor nests. They express CD4 and all markers of human Tregs. As observed for conventional CD4+ T cells, Ti-Tregs show central-memory and effector-memory phenotype. Based on the expression of activation, immunosuppression and ICP (gene and protein level), Tregs exhibit a fully activated phenotype in tumor compared to distant sites of NSCLC patients. Interestingly, Ti-Tregs display a common phenotype in TLS versus out TLS but with some key specificities regarding immune checkpoint expression.

In conclusion, the presence of several Ti-Treg subsets in lung tumor suggests they may use distinct mechanisms to exert their immunosuppressive functions in the different tumor areas.

Citation Format: Priyanka DEVI, Sylvain Leveugle, Jeremy Goc, Helene Kaplon, Claire Germain, Samantha Knockaert, Pierre Validire, Diane Damotte, Sandrine Katsahian, Wolf Herve Fridman, Catherine Sautes-Fridman, Myriam Lawand, Marie-Caroline Dieu-Nosjean. Identity card of tumor-infiltrating regulatory T cells in the context of tertiary lymphoid structures in lung cancer patients. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-273.
Abstract LB-276: The identification and characterization of prostate adenocarcinoma tumor initiating cells
Authors: Agarwal Supreet , Hynes Paul , Lake Ross , Fang Lei , Tillman Heather , Beshiri Mike , Jansson Keith , Karthaus Wouter , Iaquinta Philip , Sawyers Charles , Kelly Kathleen
Published: July 2016

Primary prostate cancer is typically of luminal phenotype. However, little is known about the stem/progenitor properties of transformed luminal tumor cells as they fail to survive in culture. Using the organoid culture methodology, we show two distinct luminal progenitors in aggressive Pten/Tp53-null mouse model of prostate cancer. Not only did tumors contain previously described multipotent progenitors, but also a major population of committed luminal progenitors. The distinction between committed luminal and multipotent organoids was also evident in subcutaneous grafts as tumors of adenocarcinoma or multilineage histological phenotypes were observed, respectively. Moreover, using organoids we show that the self-renewing capacity of luminal-committed progenitors is a tumor-specific property, absent in benign luminal cells. Further, a significant fraction of luminal progenitors displayed resistance to in vivo castration as well as to androgen receptor inhibition ex vivo. Importantly, 3D organoid techniques have allowed us to relate our findings in humans as we can successfully grow similar luminal tumor populations from patient derived xenografts (PDXs) models of prostate cancer. In all, these data reveal two distinct luminal tumorigenic populations in mouse models of prostate cancer, providing insight into luminal tumor initiating cells in prostate cancer that can also influence response to therapy.

Citation Format: Supreet Agarwal, Paul Hynes, Ross Lake, Lei Fang, Heather Tillman, Mike Beshiri, Keith Jansson, Wouter Karthaus, Philip Iaquinta, Charles Sawyers, Kathleen Kelly. The identification and characterization of prostate adenocarcinoma tumor initiating cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-276.
Abstract LB-278: Cancer stem cells as the novel origin of cancer-associated fibroblast-like cells
Authors: Nair Neha , Vaidyanath Arun , Hoshikawa Kenta , Sanchez Calle Anna , Kasai Tomonari , Seno Masaharu
Published: July 2016

Cancer associated fibroblast (CAF) is an indispensible component of stromal microenvironment mediating cancer progression notably in breast, prostate and pancreatic carcinoma. CAF heterogeneity is attributed to its possible multiple origin majorly from tumor resident fibroblast, mesenchymal stem cells, epithelial cells. The present work analyses the possibility of cancer stem cells (CSC) as the source of CAF like cells. For this, we have used the model of CSC established in our laboratory derived from mouse induced pluripotent cells (miPSCs) by culturing with conditioned medium from cancer cell lines mimicking tumor microenvironment. In the present study, human breast cancer derived T47D and BT549 cells have been cultured to collect the conditioned medium. Since the green fluorescent protein (GFP) gene was integrated under the control of Nanog promoter in the miPSCs used in this study, the resulting CSCs was assessed by the expression of GFP for the undifferentiated state. The differentiation of CSCs was tracked by the attenuation of GFP.

The CSC population was multiplied by serial subcutaneous and orthotropic transplantation in nude mice. This population was further enriched by sphere formation. Then these spheres were allowed to differentiate into adhesive cells. The CAF-like cells were separated from the differentiated population. The expression of prominent CAF markers fibroblast specific protein (FSP), fibroblast activation protein (FAP), vimentin, collagen type1 α1, and smooth muscle actin (SMA) were assessed by real time PCR. Immuno-cytochemistry of the cells for the expression of FSP, FAP and SMA was further confirmed for the evidence that cancer stem cells should be a possible source of CAF. This model should elaborate the functional heterogeneity amongst CAFs and help design therapeutic interventions to inhibit such conversions.

Citation Format: Neha Nair, Arun Vaidyanath, Kenta Hoshikawa, Anna Sanchez Calle, Tomonari Kasai, Masaharu Seno. Cancer stem cells as the novel origin of cancer-associated fibroblast-like cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-278.
Abstract LB-285: Bioinformatic identification of prognostic signature defined by copy number alteration and expression of CCNE1 in non-muscle invasive bladder cancer
Authors: Chu In-Sun , Leem Sun-Hee
Published: July 2016

Because non-muscle-invasive bladder cancer (NMIBC) is clinically heterogeneous, predicting disease progression for patients with bladder cancer still remains a major clinical challenge, despite numerous investigations for NMIBC. In this study we identify a prognostic signature to predict the heterogeneity of NMIBC using an integrative analysis of copy number and gene expression data. We analyzed the copy number and gene expression profile data of bladder cancer obtained from The Cancer Genome Atlas (TCGA) consortium. A prognostic signature based on copy number altered gene was developed and validated in three independent patient cohorts (n = 501). Diverse statistical methods were applied to identify and validate a signature. Of the fourteen significant copy number altered molecules reported previously, ten were significantly correlated between copy number and expression changes in the TCGA dataset. Among them, CCNE1 and its co-expressed genes were significantly associated with disease progression, which was validated in the independent cohorts. The CCNE1 signature was shown to be an independent risk factor by a multivariate analysis (HR 4.99, 95% CI = 1.054 to 23.659, P = 0.043). Lastly, gene network and upstream regulator analyses revealed that NMIBC progression might be mediated by CCND1-CCNE1-SP1 pathways. The prognostic molecular signature defined by copy number and expression changes of CCNE1 suggests a novel diagnostic tool for predicting the likelihood of progression in NMIBC.

Citation Format: In-Sun Chu, Sun-Hee Leem. Bioinformatic identification of prognostic signature defined by copy number alteration and expression of CCNE1 in non-muscle invasive bladder cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-285.
Abstract LB-288: Investigation on the interaction between human beta defensin type 3 (hBD-3) and ectodomain of EphA2 receptor using molecular dynamics simulation method
Authors: Zhang Liqun
Published: July 2016

Defensins are a family of cationic cysteine-rich small molecules. In general, human beta defensins (hBD) are produced by epithelial cells, and they are important to the human innate immune system. Experimental results showed that defensins can directly bind with receptors such as chemokine receptors and act through several mechanisms that may trigger antitumor immunity or facilitate the host's immune response (or evasion) and promote tumor progression. hBD type 3 (hBD-3) is newly discovered and is unique that it has a net charge density much higher than other family members, and has the ability to undergo oligomerization. On the other hand, EPH receptors and their corresponding ligands (ephrins) are membrane-bound proteins that require direct cell-cell interactions for the activation of the receptors. Eph-ephrin system plays a critical role in tumor and vascular functions during carcinogenesis. Once EPH receptor A2 (EphA2) is ligand-independent, it can over-express itself in many types of tumors, and behave like an oncogene. Besides, many studies suggested that mutations that affect EPH receptor function play a role in cancer initiation or progression. Therefore, the EphA2-ephrin5 system can be a novel target for the development of anti-tumorigenic and anti-angiogenic therapies, and understanding its interaction with other molecules is important to demystify the cancer mechanism.

Both EphA2 ectodomain (ECD) and hBD-3 exist outside of the cell membrane. In order to find out the interaction mechanism of hBD-3 with EphA2 receptor, initially blind docking using Cluspro software was performed to predict all the possible binding sites between hBD-3 oligomer and EphA2 ECD in the free and bound forms. After that, all atom molecular dynamics CHARMM simulations were performed to find out the most probable binding interfaces between hBD-3 oligomer and EphA2 ECD. It was found that hBD-3 has multiple stable binding sites on the EphA2 ECD, and its binding on the ligand-binding domain (LBD) of EphA2 could influence the binding of ephrin5 with EphA2. Those suggest the hBD-3 could play a role in cancer initiation.

Citation Format: Liqun Zhang. Investigation on the interaction between human beta defensin type 3 (hBD-3) and ectodomain of EphA2 receptor using molecular dynamics simulation method. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-288.
Abstract LB-296: Multi-target sDNA increases colorectal cancer screening among previously non-compliant patients: The USMD physician services experience
Authors: Prince Mark , Lester Lynn , Chinawala Rupal , Berger Barry M.
Published: July 2016

Background: The multi-target stool DNA test (mt-sDNA) is an FDA approved, high sensitivity, colorectal cancer screening test. We evaluated mt-sDNA in non-screening-compliant Medicare patients and assessed compliance and diagnostic colonoscopy usage and correlated positive mt-sDNA results with colonoscopy findings.

Methods: We performed a retrospective medical records review (October 2014 - September 2015) to identify mt-sDNA use in average-risk Medicare eligible patients treated by USMD Physician Services (Dallas, Tx) who were not previously compliant with colon cancer screening. We offered mt-sDNA to patients who were either >10 years since last colonoscopy and/or >1 yr. since last fecal occult blood test. Follow-up colonoscopy was advised for all patients with a positive result. Colonoscopy and pathology results were reviewed with patient classification based on the most significant pathology finding (index lesion). Size, location and total number of polyps and adenomas were tabulated.

Results: Over 12 months, 77 providers ordered 393 mt-sDNA studies with 347 completed (88.3% compliance). Patient mean age was 69.8 (50-85) and patients were 64% female. Mt-sDNA was negative in 85.3% (296/347) and positive in 14.7% (51/347). Follow-up colonoscopy was performed in 46 positive patients (90.2% colonoscopy compliance) with 3 patients refusing colonoscopy and two patients lost to follow up. Index findings included: colon cancer (4/46, 8.7%), advanced adenomas (21/46, 45.6%), non-advanced adenoma (9/46, 19.6%), and negative results (12/46,23.0%). The positive predictive value for advanced colorectal lesions was 54.3% and for any colorectal neoplasia was 73.9%.

Conclusions: The availability of mt-sDNA provided significant medical benefit to our previously screening non-compliant Medicare population. Patients with clinically critical advanced colorectal neoplasia were identified in this cohort due to high compliance with both mt-sDNA screening and follow-up diagnostic colonoscopy.

Citation Format: Mark Prince, Lynn Lester, Rupal Chinawala, Barry M. Berger. Multi-target sDNA increases colorectal cancer screening among previously non-compliant patients: The USMD physician services experience. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-296.
Abstract LB-310: NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 4-like 2 (NDUFA4L2) reduces oxidative stress to promote hepatocellular carcinoma
Authors: Lai Robin Kit-Ho , Xu Irix Ming-Jing , Chiu David Kung-Chun , Tse Aki Pui-Wah , Wei Larry Lai , Law Cheuk-Ting , Lee Derek , Wong Chun-Ming , Wong Maria Pik , Ng Irene Oi-Lin , Wong. Carmen Chak Lui
Published: July 2016

Background & Aims: Liver is a major metabolic organ, yet the detailed metabolic alterations driving hepatocellular carcinoma (HCC) remain elusive. The rapid growing nature of HCC results in oxygen deprivation or hypoxia in regions of tumors with insufficient blood supply. Hypoxia unbalances the electron flow through the electron transport chain (ETC) resulting in reactive oxygen species (ROS) accumulation. Here we aim at delineating the mechanisms by which HCC evades oxidative stress. Methods: We performed transcriptome sequencing to study the gene expression profile in both HCC patients and HCC cell line. The mRNA expression of 100 paired HCC and corresponding non-tumorous tissues were analyzed. Stable RNAi knockdown by shRNA and genetic knockout by TALEN were established in HCC cells for functional characterization. Results: We demonstrated that HCC cells specifically utilized the mitochondrial protein NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 4-like 2 gene (NDUFA4L2), in the complex I of the ETC, to survive hypoxia. NDUFA4L2 was drastically over-expressed in human HCC and closely associated with poor clinical outcomes in HCC patients. We confirmed that NDUFA4L2 was regulated by HIF-1α in HCC cells. Inactivation of HIF-1α/NDUFA4L2 in different HCC cell lines increased mitochondrial activity and oxygen consumption, resulting in ROS accumulation and ROS-mediated apoptosis in HCC cells. Knockdown of NDUFA4L2 markedly suppressed HCC growth and metastasis in vitro and in vivo. In addition, HIF inhibitors, digoxin and sorafenib, significantly suppressed growth of tumors that expressed high level of NDUFA4L2 in orthotopic HCC model. Conclusions: Our results have unprecedentedly uncovered the clinical relevance and oncogenic roles of NDUFA4L2 in HCC.

Citation Format: Robin Kit-Ho Lai, Irix Ming-Jing Xu, David Kung-Chun Chiu, Aki Pui-Wah Tse, Larry Lai Wei, Cheuk-Ting Law, Derek Lee, Chun-Ming Wong, Maria Pik Wong, Irene Oi-Lin Ng, Carmen Chak Lui Wong. NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 4-like 2 (NDUFA4L2) reduces oxidative stress to promote hepatocellular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-310.
Abstract LB-313: Upregulation of lactate dehydrogenase A by 14-3-3ζ leads to increased glycolysis critical for breast cancer initiation and progression
Authors: Chang Chia-Chi , Zhang Chenyu , Sahin Ozgur , Zhang Qingling , Wang Hai , Zhang Jian , Rehman Sumaiyah , Li Ping , Yu Dihua
Published: July 2016

Metabolic reprogramming is a hallmark of cancer. Elevated glycolysis in cancer cells switches the cellular metabolic flux to produce more biological building blocks, thereby facilitating cellular adaptation to the adverse environment and sustaining rapid proliferation. Recently, new evidence has emerged that abnormal metabolic reprogramming may occur at the early-stages of neoplasia and critically contribute to early transformation and cancer initiation. Previously, we reported that 14-3-3ζ overexpression is an early event in human breast cancer, taking place as early as in the atypical ductal hyperplasia (ADH) stage. We therefore hypothesized that 14-3-3ζ overexpression in breast epithelial cells may trigger metabolic reprogramming by elevating glycolysis that contributes to cancer initiation. In the present study, our bioinfomatics analysis of microarray profiling data from the early-stages breast neoplastic lesions revealed that 14-3-3ζ expression is strongly correlated with the expression of genes of the canonical glycolytic pathway, particularly lactate dehydrogenase A (LDHA). Experimentally, increasing 14-3-3ζ expression in human non-transformed mammary epithelial cells (hMECs), MCF10A and MCF12A, transcriptionally up-regulated LDHA expression, elevated glycolytic activity, and promoted early transformation of hMECs. Conversely, knockdown of LDHA in the 14-3-3ζ-overexpressing hMECs significantly reduced glycolytic activity and inhibited transformation. Together, our data demonstrate that overexpression of 14-3-3ζ in early stage pre-cancerous breast epithelial cells may trigger an elevated glycolysis by activating the MEK-ERK-CREB signaling axis and transcriptionally up-regulating LDHA, thereby critically contributes to human breast cancer initiation.

Citation Format: Chia-Chi Chang, Chenyu Zhang, Ozgur Sahin, Qingling Zhang, Hai Wang, Jian Zhang, Sumaiyah Rehman, Ping Li, Dihua Yu. Upregulation of lactate dehydrogenase A by 14-3-3ζ leads to increased glycolysis critical for breast cancer initiation and progression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-313.
Abstract LB-320: Obligate lesions and diverse evolutionary patterns drive small cell bladder cancer
Authors: Penson Alexander , Chang Matthew T. , Desai Neil , Socci Nicholas D. , Shen Ronglai , Seshan Venkatraman E. , Kundra Ritika , Abeshouse Adam , Viale Agnes , Cha Eugene K. , Bochner Bernard H. , Rosenberg Jonathan E. , Bajorin Dean F. , Schultz Nikolaus , Berger Michael F. , Iyer Gopa , Solit David B. , Al-Ahmadie Hikmat , Taylor Barry S.
Published: July 2016

Small cell bladder cancer (SCBC) is a rare and aggressive neuroendocrine tumor with a dismal prognosis and limited treatment options. As SCBC is histologically indistinguishable from small cell lung cancer, a shared pathogenesis and cell of origin has been proposed. To determine whether SCBCs arise from a pre-existent urothelial carcinoma or share a molecular pathogenesis in common with small cell lung cancer, we performed integrative analysis of 61 SCBC patients to identify histology- and organ-specific similarities and differences with urothelial carcinomas and small cell lung cancers. SCBC has a high somatic mutational burden driven predominantly by an APOBEC-mediated mutational process. Obligate mutations in TP53, RB1, or the TERT promoter were present in nearly all samples. While these events arose early in all affected tumors, reflecting an evolutionary branch point that drove small cell lineage differentiation, they were not the founding transforming event, as they were often preceded by diverse and less common driver mutations many of which are common in bladder urothelial cancers but not small cell lung tumors. The majority of patient tumors (72%) also underwent whole-genome duplication (WGD). While arising at different chronological points in the evolution of the disease, WGD was often preceded by biallelic mutations in TP53 with retention of two intact copies. Together, our findings indicate that small cell cancers of the bladder and lung have a convergent but distinct pathogenesis and SCBCs likely arise from a cell of original shared with urothelial bladder cancer.

Citation Format: Alexander Penson, Matthew T. Chang, Neil Desai, Nicholas D. Socci, Ronglai Shen, Venkatraman E. Seshan, Ritika Kundra, Adam Abeshouse, Agnes Viale, Eugene K. Cha, Bernard H. Bochner, Jonathan E. Rosenberg, Dean F. Bajorin, Nikolaus Schultz, Michael F. Berger, Gopa Iyer, David B. Solit, Hikmat Al-Ahmadie, Barry S. Taylor. Obligate lesions and diverse evolutionary patterns drive small cell bladder cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-320.
Abstract LB-339: Mutation analysis of cell-free DNA captures heterogeneity of individual circulating tumor cells in metastatic breast cancer
Authors: Guttery David S. , Shaw Jacqueline A. , Hills Allison , Fernandez-Garcia Daniel , Page Karen , Rosales Brenda , Goddard Kate , Hastings Robert , Luo Jinli , Ogle Olivia , Woodley Laura , Ali Simak , Stebbing Justin , Coombes Charles
Published: July 2016

Circulating tumor cells (CTCs) are more technically challenging to isolate and characterize than circulating cell-free DNA (cfDNA). However, analysis of single CTCs may reveal more actionable information to guide targeted therapies than mutation tracking in cfDNA. Here, we compared CTC count with cfDNA levels in 113 patients with metastatic breast cancer. In 5 patients we compared mutations in 40 individual CTCs and matched cfDNA using targeted amplicon sequencing of 54 genes (∼2800 COSMIC mutations) at >2500x mean read depth. Total cfDNA levels were significantly associated with overall survival (hazard ratio (HR), 2.2; 95% CI, 1.1 to 4.3; P = 0.021) and correlated with CTC counts (P = 0.002). Four patients had mutations detected in cfDNA, the levels of which tracked with levels of circulating tumor DNA (ctDNA). Mutational heterogeneity was seen across individual CTCs (including mutations in PIK3CA, ESR1 and KRAS) and was reflected in matched cfDNA, but cfDNA had other mutations not seen in CTCs (including TP53). Therefore, cfDNA reflects persisting CTCs and enables monitoring of the metastatic burden for clinical decision-making when CTCs cannot be obtained. Moreover, monitoring of total cfDNA levels could provide an inexpensive and simple measure of treatment response.

Citation Format: David S. Guttery, Jacqueline A. Shaw, Allison Hills, Daniel Fernandez-Garcia, Karen Page, Brenda Rosales, Kate Goddard, Robert Hastings, Jinli Luo, Olivia Ogle, Laura Woodley, Simak Ali, Justin Stebbing, Charles Coombes. Mutation analysis of cell-free DNA captures heterogeneity of individual circulating tumor cells in metastatic breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-339.
Abstract LB-358: Plasma HGF level is associated with obesity, increased risk and reduced survival of pancreatic cancer
Authors: Yan Dong , Chang Ping , Li Donghui
Published: July 2016

Background: Obesity is one of the modifiable risk factors for the development and progression of pancreatic cancer. However, the mechanisms of obesity-related pancreatic cancer has not been well defined. In our previous analysis of the genome-wide association data, we identified a significant interaction of obesity with cytokine signaling pathway. Methods: In the current study, we selected several genes from this pathway that code for secreted or membrane proteins and measured the protein levels in plasma samples of 100 pancreatic patients and 100 healthy controls drawn from a case-control study conducted at MD Anderson. Cases and controls were frequency matched by age, sex, race and body mass index (BMI). Plasma levels of HGF (hepatocyte growth factor), MIP-1β (microphage infiltration factor 1-beta), FasL (Fas ligand) and PLA2 (phospholipase A2) were measured using the Luminex and ELISA methods. The protein levels were analyzed in relation to cancer risk and patient survival using analysis of variance, non-parametric test, Kaplan Meier plot, log-rank test, and Cox proportional hazard regression models. Results: A significantly higher level of HGF and MIP-1β and lower level of FasL was observed in cancer patients than in controls, with the mean values of 365.7 vs. 146.2 pg/ml, P< 0.001; 216.5 vs.122.3 pg/ml, P = 0.002, 23.1 versus 29.4 pg/ml, P<0.001, respectively. Individuals with a high HGF or low FasL level had, respectively, 4.6-fold (95% confidence interval [CI] = 2.4-9.1, P<0.001) and 2.9-fold (95% CI = 1.55-5.38, P = 0.001) increased risk of pancreatic cancer with adjustment for other known risk factors. Furthermore, a significant association of HGF level with BMI was observed in cases (R2 = 0.269, P = 0.005). The HGF level (mean ± SE pg/ml) was significantly higher in obese (682±204) than that in overweight (268±40) or normal weight patients (313±71), P = 0.01 and 0.005, respectively. Patients with higher levels of HGF had a significantly reduced overall survival than those with lower levels (9.3 vs. 13.3 months, P = 0.006). Cox proportional hazard regression analysis demonstrated that a higher level of HGF was an independent prognostic factor (Hazard Ratio = 2.61, 95% CI = 1.22-5.64, P = 0.01) after adjusting for BMI, disease stage and other clinical factors. Conclusion: A higher plasma level of HGF was related to obesity and overall survival in pancreatic adenocarcinoma, which may serve as a potential diagnostic and prognostic marker for pancreatic cancer.

Citation Format: Dong Yan, Ping Chang, Donghui Li. Plasma HGF level is associated with obesity, increased risk and reduced survival of pancreatic cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-358.
Abstract LB-364: Natural history of age-related lobular involution and impact on breast cancer risk
Authors: Radisky Derek , Visscher Daniel W. , Frank Ryan , Vierkant Robert A. , Winham Stacey J. , Stallings-Mann Melody L. , Hoskin Tanya L. , Nassar Aziza , Vachon Celine M. , Denison Lori A. , Hartmann Lynn C. , Frost Marlene H. , Degnim Amy C.
Published: July 2016

Background: Age-related lobular involution (LI) is a physiological process in which the terminal duct lobular units of the breast regress as a woman ages. Analyses of breast biopsies from women with benign breast disease (BBD) have found that extent of LI is negatively associated with subsequent breast cancer development. Here we assess the natural course of LI within individual women, and the impact of progressive LI on breast cancer risk.

Methods: The Mayo Clinic BBD cohort consists of 13,455 women with BBD from 1967-2001. The BBD cohort includes 1115 women who had multiple benign biopsies, 106 of whom had developed breast cancer. Within this multiple biopsy cohort, the progression of the LI process was examined by age at initial biopsy and time between biopsies. The relationship between LI progression and breast cancer risk was assessed using standardized incidence ratios (SIRs) and by Cox proportional hazards analysis.

Results: Women who had multiple biopsies were younger age and had a slightly higher family history of breast cancer as compared with the overall BBD cohort. Extent of LI at subsequent biopsy was greater with increasing time between biopsies and for women age 55+ at initial biopsy. Among women with multiple biopsies, there was a significant association of higher breast cancer risk among those with involution stasis (lack of progression, HR 1.63) as compared with those with involution progression, p = 0.036.

Conclusions: The multiple biopsy BBD cohort allows for a longitudinal study of the natural progression of LI. The majority of women in the multiple biopsy cohort showed progression of LI status between benign biopsies, and extent of progression was highest for women who were in the perimenopausal age range at initial biopsy. Progression of LI status between initial and subsequent biopsy was associated with decreased breast cancer risk.

Citation Format: Derek Radisky, Daniel W. Visscher, Ryan Frank, Robert A. Vierkant, Stacey J. Winham, Melody L. Stallings-Mann, Tanya L. Hoskin, Aziza Nassar, Celine M. Vachon, Lori A. Denison, Lynn C. Hartmann, Marlene H. Frost, Amy C. Degnim. Natural history of age-related lobular involution and impact on breast cancer risk. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-364.
Abstract LB-366: miRNA as markers of CIN risk and persistence for optimization of HPV-based cervical cancer screening
Authors: Naccarati Alessio , De Maria Daniela , Cordero Francesca , Pardini Barbara , Gouzounis Stavrula , Arigoni Maddalena , Rizzolo Raffaella , De Marco Laura , Gillio-Tos Anna , Vineis Paolo , Ronco Guglielmo
Published: July 2016

Cervical cancer (CC) is caused by oncogenic human papillomaviruses (HR-HPVs) infection and exhibits well-defined clinical stages associated with different steps of tumorigenesis. Screening for CC based on HPV is more effective than cytology, but it may increase costs and over treatment of spontaneously regressive high-grade lesions (hgCIN). Thus triage of HPV positive women is needed. Several microRNAs (miRNAs) are dysregulated in CC and hgCIN. However, the studies so far are based on miRNA candidate genes or array approach, mainly in tumor/healthy tissues. The aim of this study is to investigate, for the first time, miRNA profiles by next-generation sequencing in exfoliated cervical cells of HPV-positive women in relation to the presence of hgCIN lesions or as predictors of persistence/progression of CIN lesions. The study is nested in a large Italian multi-centre randomised controlled trial recruiting women in population-based screening programs that actively invite women aged 25-64 years. The study population includes HPV-positive women with CIN2 and CIN3 or without hgCIN lesion. For the discovery phase libraries of 100 samples of exfoliated cervical cells have been set up for miRNA sequencing. For the validation an additional set of 250 samples has already been selected.

Preliminary results shows that, after correction for multiple testing, 44 miRNAs resulted dysregulated in women with vs. those without prevalent hgCIN. This set includes some miRNAs previously reported in the literature, such as miR-100 and miR-126, and other additional miRNAs not included in arrays before. The association of miRNAs with occurrence of hgCIN and clearance of infection during follow up reveals other significantly differentially expressed miRNAs under investigation for their biological roles. The high stability of miRNAs, in contrast to the fast degradation of mRNA and proteins, allows their accurate determination also in samples of exfoliated cervical collected during screening. Validation of the results, associations with other investigated markers and with HPV genotypes will be performed.

Study supported by Italian Association on Cancer Research (AIRC, IG2013 N.14119).

Citation Format: Alessio Naccarati, Daniela De Maria, Francesca Cordero, Barbara Pardini, Stavrula Gouzounis, Maddalena Arigoni, Raffaella Rizzolo, Laura De Marco, Anna Gillio-Tos, Paolo Vineis, Guglielmo Ronco. miRNA as markers of CIN risk and persistence for optimization of HPV-based cervical cancer screening. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-366.
Abstract LB-370: Fine mapping of 32 breast cancer risk loci in women of East Asian ancestry
Authors: Zeng Chenjie , Cai Qiuyin , Long Jirong , Zheng Ying , Xiang Yong-Bing , Shi Jiajun , Shu Xiao-Ou , Zheng Wei
Published: July 2016

Background: Genome-wide association studies (GWAS) have identified multiple genetic susceptibility loci for breast cancer risk. The majority of these loci, however, were identified in studies conducted in European descendants. A comprehensive evaluation of these risk loci using densely genotyped data in East Asians may help to identify independent association signals and risk variants that are more closely related to breast cancer risk in East Asian women than those reported initially.

Methods: We performed a fine-scale mapping of 32 breast cancer susceptibility loci identified in previous GWAS in 5,703 breast cancer cases and 5,766 controls in studies conducted in Shanghai, China, Nearly 4,100 variants in these 32 loci were genotyped. We imputed genotypes using the 1000 Genomes Project (Phase 3) as the reference. Included in this analysis were 4,086 genotyped and more than 17,000 well-imputed common variants. At each locus, analyses using logistic regression models were conducted to identify variants that better capture the association with breast cancer risk and new risk variants that are not correlated with the risk variants reported from the initial studies (index SNPs). To identify additional independent risk variants, logistic regression conditional on the most significantly associated variant was performed.

Results: We identified risk variants that better captured the association than the index SNPs at 3 loci (2q35, rs1478596, OR = 1.14, P = 5.6×10-6; 6q25.1, rs9322344, OR = 0.89, P = 3.1×10-5;11q24.3, rs11221769, OR = 1.12, P = 6.1 ×10-5), new risk variants at 2 loci (1p13.2, rs3789614, OR = 1.31, P = 0.002; 2q31.1, rs117055746, OR = 0.87, P = 0.004), and independent risk variants at two loci (6q14.1, rs200626240, OR = 0.89, P = 0.001; 6q25.1 rs2982684, OR = 0.87, P = 2.7×10-5, Chr6:152107810, OR = 0.91, P = 1.4×10-4).

Conclusion: This study identified multiple risk variants that were more relevant to East Asians than the index SNPs. These risk variants combined explained a further 2% of the familial relative risk in this population. This study highlighted the need of fine-mapping studies in non-European ancestral populations to search for additional risk variants.

Citation Format: Chenjie Zeng, Qiuyin Cai, Jirong Long, Ying Zheng, Yong-Bing Xiang, Jiajun Shi, Xiao-Ou Shu, Wei Zheng. Fine mapping of 32 breast cancer risk loci in women of East Asian ancestry. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-370.
Abstract LB-378: Ankyrin repeat domain 30A as a novel molecular marker of columnar alterations in benign breast disease and subsequent breast cancer in African American women
Authors: Holowatyj Andreana N. , Ali-Fehmi Rouba , Ruterbusch Julie J. , Abdulfatah Eman , Pardeshi Vishakha , Roquiz Woodlyne , Alosh Baraa , Bandyopadhyay Sudeshna , Radisky Derek , Cote Michele L.
Published: July 2016

An estimated 1.6 million breast biopsies are performed annually in the United States, and the vast majority are benign. Benign breast disease is an established breast cancer risk factor for both Caucasian and African American women. Yet little is still known about benign breast tissue from African American women, who suffer from disproportionately high-grade tumors and poorer breast cancer survival. To better characterize the risk of breast cancer among African American women with benign breast disease, we assessed benign breast disease pathology of African American women in our metropolitan Detroit cohort (n = 3,737). We classified benign breast lesions for pathological characteristics and used logistic regression to estimate subsequent breast cancer risk with histological characteristics. We observed that women whose biopsies showed columnar alterations, columnar cell lesions of the breast, were 1.78-fold more likely to develop an invasive breast cancer [odds ratio (OR) 1.78, 95% confidence interval (CI) 1.19-2.65] after adjusting for age at biopsy, biopsy year, and overall impression. Columnar alterations were not found to be significantly associated with risk of developing ductal carcinoma in situ (DCIS) among African American women [OR 1.53, 95% CI 0.83-2.82]. Next, to identify molecular markers associated with columnar alterations in benign breast lesions, we performed gene expression analyses on biopsy tissues from 27 African American women that subsequently developed an invasive breast cancer. Transcriptional profiling of these lesions identified differentially expressed genes significantly associated with columnar alterations, including ankyrin repeat domain 30A (ANKRD30A). ANKRD30A, also known as NY-BR-1, is a mammary tissue-specific differentiation antigen that is overexpressed in breast cancers. Increased expression of ANKRD30A in benign breast lesions with columnar alterations may be indicative of subsequent breast cancer development. Taken together, novel molecular markers, including ANKRD30A, and ethnicity can lead to better risk assessment and earlier diagnosis of breast cancer.

Citation Format: Andreana N. Holowatyj, Rouba Ali-Fehmi, Julie J. Ruterbusch, Eman Abdulfatah, Vishakha Pardeshi, Woodlyne Roquiz, Baraa Alosh, Sudeshna Bandyopadhyay, Derek Radisky, Michele L. Cote. Ankyrin repeat domain 30A as a novel molecular marker of columnar alterations in benign breast disease and subsequent breast cancer in African American women. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-378.
Abstract PL03-02: The AML genome(s)
Authors: Ley Timothy J.
Published: July 2016

Acute Myeloid Leukemia (AML) is a clonal disorder of hematopoietic stem/progenitor cells. These cells have been shown to accumulate random mutations as a function of time; however, some of these mutations are non-random and can cause the expansion of the stem cells that contain them, creating a benign ‘preleukemic’ state that has recently been dubbed “Clonal Hematopoiesis of Indeterminate Potential” (CHIP). As these clones expand, they can occasionally acquire additional mutations that create a profound advantage for these cells, yielding an AML “ founding clone”. Founding clones generate subclones that have unique properties that are probably defined by the specific mutations that they contain. These subclones, found in virtually all AML cases, shape the responses of AML patients to therapy, and often contribute to relapse. Clonal heterogeneity is therefore a hallmark of AML at presentation, and approaches to assess risk of relapse must take this into account. Further, approaches to cure this disease will probably require successful targeting of the mutations that initiate the disease, which are the only ones that are found in all the cells within the tumor. Frequently, the initiating mutations that expand the preleukemic clone are persistent when patients are in clinical remission, strongly suggesting that these events are important for relapse. The precise mechanisms by which canonical initiating events predispose to AML are largely unknown, but many of these mutations do cause significant changes in the epigenome that are probably relevant for pathogenesis.

Citation Format: Timothy J. Ley. The AML genome(s). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr PL03-02.
Abstract SY04-01: The in vivo gain-of-function activities of mutant p53
Authors: Lozano Guillermina
Published: July 2016

The p53 tumor suppressor is the most often altered gene in human cancers with missense mutations in the DNA binding domain accounting for approximately 70% of alterations. Mutant p53 proteins have gain-of-function activates that contribute to more aggressive tumors with increased metastatic properties. Mechanisms by which mutant p53 exerts its gain of function will be discussed. In addition, I will discuss the generation of a novel mouse model that allows Cre-mediated expression of mutant p53 in specific cell types leaving the surrounding stroma and immune system wild type for p53 as occurs in human somatic tumors.

Citation Format: Guillermina Lozano. The in vivo gain-of-function activities of mutant p53. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr SY04-01.
Abstract SY09-01: The tumor immunity continuum as a framework for rational combinations
Authors: Hegde Priti S.
Published: July 2016

The PD-L1/PD-1 pathway is emerging as a central axis for immune-based therapeutic strategies in the treatment of multiple cancers. While these therapies are defined by durable responses extending to impressive overall survival improvement, this clinical benefit is limited to a few. We utilized gene signature analysis of cell subsets associated with the TME and molecular classification of disease using whole transcriptome analysis, PD-L1 IHC, CD8 IHC and tumor mutational load to identify signatures associated with immune response to atezolizumab (ati-PDL1) therapy from large Phase II trials and describe signatures associated with immune escape. Namely, we demonstrate that a preexisting immune response associated with inflamed tumors is highly predictive of clinical benefit to checkpoint inhibition. In addition, we describe both histological and molecular phenotypes associated with immune escape. The biological drivers of PD-L1 expression on tumor cells and tumor infiltrating immune cells will be presented. Using the lessons learned from human tumor immunology, we propose the concept of the tumor immunity continuum as a framework to expand the patient pool that derive long-term clinical benefit by harnessing their immune system.

Citation Format: Priti S. Hegde. The tumor immunity continuum as a framework for rational combinations. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr SY09-01.
Abstract SY38-02: Immune-related changes in breast tumor evolution
Authors: Polyak Kornelia
Published: July 2016

Increasing evidence supports a role for the immune system in tumor initiation and progression. Immune cells shape tumor evolution directly (e.g., anti-tumor immune response) and indirectly (e.g., changing the microenvironment) by selecting for cancer cells with certain properties. Immunotherapy is one of the most promising current therapeutic options and almost the only one that can effectively cure metastatic disease. However, thus far its success has been limited to a subset of patients. This may be a consequence of heterogeneity among tumors for mechanisms by which they escape immune surveillance. We hypothesized that the in situ to invasive carcinoma transition is a critical tumor progression step for immune escape in breast cancer that defines subsequent tumor evolution. This is due to the fact that in DCIS (ductal carcinoma in situ) cancer cells are still physically separated from the stroma by the basement membrane and myoepithelial cell layer, and tumor-infiltrating leukocytes are rarely detected in direct contact with cancer cells. In contrast, with invasive progression, cancer cells and leukocytes are intermingled, thus, only cancer cells that can survive in this environment will play a role in disease progression. To begin dissecting mechanisms of immune escape during breast tumor progression, we analyzed the composition of leukocytes in normal breast tissues, DCIS, and invasive ductal carcinomas (IDC) by polychromatic FACS and found significant changes in multiple cell types. We also analyzed the gene expression profiles of T cells from each of these tissue types and found that T cells in DCIS are different from those in normal breast and invasive tumors. We validated some of these changes by multicolor immunofluorescence. We also performed high throughput sequencing of T cell receptor (TCR) of T cells in DCIS and found potential clonal expansions shared among patients. Lastly, we performed multicolor immunofluorescence analysis of checkpoint proteins in DCIS and IDC of different subtypes, and detected significant differences in expression patterns. Our results imply that analyzing immune-related changes during DCIS to IDC transition will identify novel targets for immunotherapy in breast cancer.

Citation Format: Kornelia Polyak. Immune-related changes in breast tumor evolution. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr SY38-02.