bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1125: Developmental pluripotency associated 4 oncogene interacts with Wnt/β-catenin signaling and stem cells regulatory genes to control pituitary tumor cells invasiveness
Authors: Sarkar Dipak K. , Jabbar Shaima , Gangisetty Omkaram
Published: August 2018

The developmental pluripotency associated 4 (DPPA4) gene has an important role in self-renewal and pluripotency in embryonic stem cells. It is re-expressed in several malignant tumors and is identified as a new pluripotency-related oncogene. We have recently identified elevated expression of DPPA4 in aggressive pituitary tumors induced by estrogen in fetal alcohol exposed rats. In this study, we determined the role of DPPA4 in pituitary tumor growth and invasiveness by employing CRISPR/Cas9 knock-down of DPPA4 and conducting cell proliferation, in vitro soft agar anchorage-independent growth assays, and in vivo tumor formation assay using cell spheres from pituitary tumors of alcohol-fed and control-fed animals. Pituitary tumor cells of fetal alcohol-fed rats and control diet-fed rats were grown in ultra-low attachment plate to develop pituispheres. In vitro and in vivo studies showed pituispheres of alcohol-fed rats had higher cell proliferation rate, increased colony formation in soft agar, and higher tumor development in mice xenografts compared to those in pituispheres of alcohol-fed rats. Pituispheres of alcohol-fed rats also had increased mRNA levels of cell growth, cell stemness and EMT regulatory genes. CRISPR-Cas9 knockdown of DPPA4 in pituispheres reduced the tumor cell ability to proliferate, migrate and form colony in soft agar and grow in mice xenografts. Pituispheres of alcohol-fed rats also showed increased protein levels of Wnt/beta-catenin signaling molecules (Wnt 3, beta-catenin, LPR5, GSK3), stemness regulatory factors (KLF4, Sox2, Nanog) and EMT factors as compared to those in control-fed pituispheres. DPPA4 knockdown reduced the levels of Wnt signaling and stem cells regulatory proteins and EMT factors in alcohol-fed rat pituispheres. These data suggest that DPPA4 is expressed in aggressive pituitary tumors and regulates the growth and invasion of these tumors via activation of Wnt/beta-catenin signaling and stem cells regulatory genes. (This work is supported by a National Institute of Health grant R01 AA11591).

Citation Format: Dipak K. Sarkar, Shaima Jabbar, Omkaram Gangisetty. Developmental pluripotency associated 4 oncogene interacts with Wnt/β-catenin signaling and stem cells regulatory genes to control pituitary tumor cells invasiveness [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 1125.
Abstract 1128: Exosomal ID3 is pro-metastatic through guiding NRF1-induced breast cancer stem cells across the blood-brain-barrier
Authors: Das Jayanta K. , Doke Mayur , Deoraj Alok , Felty Quentin , Roy Deodutta
Published: August 2018

Nuclear respiratory factor 1 (NRF1) and inhibitor of differentiation protein 3 (ID3) are transcription regulating proteins. Recently we have uncovered a novel oncogenic function of NRF1 in breast cancer development and progression. In this study, we tested the postulate that exposure to PCB153 contributes in ID3 over-expressing endothelial stem cells (EndSCs) chaperoning and guiding of mesenchymal NRF1 breast cancer stem cells (BCSCs) across the blood-brain barrier. BCSCs/EndSCs were subjected to functional gain/loss of ID3 and/or NRF1 to test if PCB153 [1ng/ml] exposure produces NRF/ID3 signals regulating lineage specific BCSCs organ entry. First, we tested whether NRF1 promotes transmigration of breast cancer using a 3D blood-brain barrier (BBB) model consisting of breast tumor initiating cells (BTICs) or MDAMB231 cells, brain endothelial cell layer, and astrocytes. NRF1 overexpression increased the propensity for BTICs and mesenchymal enriched MDAMB231 BCSCs to adhere to brain endothelial cells and migrate across a human BBB model. Increased adhesion of NRF1+ BCSCs with ID3+ EndSCs was detected. ID3 EndSCs promoted the transmigration of BCSCs through the BBB. We found differential effects by endocrine disruptors, namely PCB153 and PCB77. The treatment with PCB153 showed increased growth of NRF1+ BCSCs tumor spheroids. PCB153 also induced the tumor cell adhesion to microvascular endothelium and transendothelial migration of BCSCs. The exosomal ID3 released from endothelial cells helped the mesenchymal NRF1+ BCSCs to cross through blood brain barriers. Xenograft experiments showed that ID3+ brain EndSCs not only supported the growth of estrogen treated NRF1+ BCSC tumorospheroids, but guided them to the brain in zebrafish. These findings show for the first time a key role for ID3 and NRF1 by which specific circulatory EndSCs accompany a particular sub-type of BCSCs to distant metastatic sites where they most likely facilitate the seeding, survival, and proliferation of BCSCs. This knowledge is important for pre-clinical testing of NRF1/ID3 modifying agents that prevent the spread of breast cancer to the brain. This work was, in part, supported by a VA MERIT Review (VA BX001463) grant to DR.

Citation Format: Jayanta K. Das, Mayur Doke, Alok Deoraj, Quentin Felty, Deodutta Roy. Exosomal ID3 is pro-metastatic through guiding NRF1-induced breast cancer stem cells across the blood-brain-barrier [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 1128.
Abstract 1136: Decrease in maturation of stem cells along the neuroendocrine lineage contributes to stem cell overpopulation that drives human CRC development
Authors: Boman Bruce M. , Zhang Tao , Modarai Shirin , Opdenacker Lynn , Jeremy Fields
Published: August 2018

Stem cell (SC) overpopulation is thought to drive tumor initiation and progression during colon cancer development, but it is unclear what causes the SC overpopulation. Because most neuroendocrine (NE) cells (NECs) in normal colonic crypts reside within the SC niche at the crypt bottom, we thought that aberrant NECs might be linked to the SC overpopulation. We hypothesized that (i) in normal crypts, SC and NEC populations are anatomically and functionally related, and (ii) during tumorigenesis, changes in these relationships quantitatively correlate with increased SC numbers. We tested this hypothesis - using quantitative immunohistochemical mapping - as a first step towards understanding how interactions between SC and NEC might become altered during colon tumorigenesis. In normal human colonic crypts, most (>80%) cells staining for ALDH1, a colonic SC marker, co-stained for chromogranin-A (CGA) and several other NE markers. Neither ALDH1+ nor CGA+ cells co-stained for MCM2, a marker for proliferating cells. These findings indicate that, in normal colonic crypts, many ALDH1+ cells have NE characteristics. In contrast, the proportion of ALDH1+ cells that co-stained for NE markers progressively decreased (to <20%) during the stepwise progression to cancer (from normal crypts to normal-appearing FAP crypts to adenomatous crypts to colon carcinomas). However, the absolute number of ALDH+ cells progressively increased during this progression, reflecting the extent of the SC overpopulation. These findings suggest that (i) NEC maturation is integral to regulation of SC population size and (ii) a decreased rate of NEC maturation contributes to SC overpopulation during colon tumorigenesis.

Citation Format: Bruce M. Boman, Tao Zhang, Shirin Modarai, Lynn Opdenacker, Fields Jeremy. Decrease in maturation of stem cells along the neuroendocrine lineage contributes to stem cell overpopulation that drives human CRC development [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 1136.
Abstract 1140: Genes preserving stem cell state in group 3 medulloblastoma brain tumor initiating cells contribute to therapy evasion and relapse
Authors: Singh Sheila Kumari , Bakhshinyan David , Venugopal Chitra , Adile Ashley , Singh Mohini , Qazi Maleeha , Manoranjan Branavan , Kameda-Smith Michelle
Published: August 2018

Medulloblastoma (MB) is the most common malignant pediatric brain tumor. Of current molecular subgroups, Group 3 patients face the highest incidence of metastatic spread and overall patient survival of less than 50%. Current clinical trials for recurrent MB patients based on genomic profiles of primary, treatment-naïve tumors provide limited clinical benefit, since recurrent metastatic MBs are highly genetically divergent from their primary tumors. By adapting the existing COG (Children's Oncology Group) protocol for children with newly diagnosed high-risk MB to the treatment of immuno-deficient mice intracranially engrafted with human MB brain tumor initiating cells (BTICs), we have characterized the rare treatment-refractory cell population in Group 3 MBs. MB cell populations recovered separately from brains and spines during the course of tumor development and therapy were comprehensively profiled for gene expression analysis, stem cell and molecular features to generate a global, comparative profile of MB cells through therapy to relapse. One of the most intriguing observations from our gene expression data was consistent over-expression in the treatment-refractory cell population of proteins belonging to the Inhibitor of DNA-binding/differentiation (ID) family (transcription factors with a basic helix-loop-helix motif that act as suppressors cellular differentiation), and a longevity-associated protein known as bactericidal/permeability-increasing fold-containing-family-B-member-4 (BPIFB4). This persistent upregulation of genes preserving undifferentiated state and cellular longevity further strengthens the hypothesis of stem-cell like cells driving tumor relapse in MB. Targeting ID1 and BPIFB4 using both knockdown (KD) and knockout (KO) strategies has resulted in decreased self-renewal and tumorigenicity of both primary and recurrent MB cells, further highlighting their potential as novel therapeutic targets in MB. Our differential genomic and gene expression profiles of the “treatment-responsive” tumors against those that fail therapy have successfully contributed to discovery and characterization of novel therapeutic targets for the most aggressive subgroup of MB.

Citation Format: Sheila Kumari Singh, David Bakhshinyan, Chitra Venugopal, Ashley Adile, Mohini Singh, Maleeha Qazi, Branavan Manoranjan, Michelle Kameda-Smith. Genes preserving stem cell state in group 3 medulloblastoma brain tumor initiating cells contribute to therapy evasion and relapse [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 1140.
Abstract 1165: A tissue-engineered bone mimetic in vitro model for monitoring metastatic PCa growth and therapy response
Authors: Paindelli Claudia , Hutmacher Dietmar , Friedl Peter , Dondossola Eleonora
Published: August 2018

Despite recent advances in prostate cancer (PCa) treatment, the outcome of metastatic disease remains frequently fatal and the underlying biology poorly understood. Thus, the development of clinically relevant in vitro models to monitor PCa biology in organotypic bone-like environment is critical to uncover mechanisms of therapy resistance and identify more effective treatments. To establish a bone-mimetic culture, we combined the following components: (i) bioactive osteoblasts depositing bone-like calcified extracellular matrix, (ii) complex 3D surface geometries, (iii) multicellular tumor application as spheroids/organoids, and (iv) applicability for live-cell microscopy to monitor the development of lesions over time. Calcified polycaprolactone (PCL) scaffolds were functionalized with bone-derived human mesenchymal stem cells (hMSCs) differentiated to osteoblasts, to generate a 3D niche-like calcified scaffold. PCa spheroids (PC3, C4-2B, patient-derived xenografts) were on-planted and their growth and invasion longitudinally monitored by advanced microscopy. PCa spheroids seeded on the organotypic bone model could be maintained and expanded over weeks, sufficient for monitoring therapy response to docetaxel, a first-line therapy for advanced PCa. The bone mimetic culture further revealed resistance to docetaxel mainly at the invasive edges, through a mechanism depending on the presence of osteoblasts. Thus, this 3D in vitro organotypic model will be suitable for dissecting the physical and molecular PCa cell-osteoblast interaction involved in PCa growth and therapy resistance.

Citation Format: Claudia Paindelli, Dietmar Hutmacher, Peter Friedl, Eleonora Dondossola. A tissue-engineered bone mimetic in vitro model for monitoring metastatic PCa growth and therapy response [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 1165.
Abstract 1168: The “humanized” chicken embryo chorioallantoic membrane (CAM) as a personalized platform for high-throughput (HTP) screening of cancer therapeutics
Authors: Shapira Shiran , Bogin Oren , Arber Nadir
Published: August 2018

Background

Every day, millions of people are taking medications that will not help them. “Personalized medicine,” is the tailoring of medical treatment to a single person, shape his response to a particular treatment and aims to better target intervention, maximize benefit and minimize harm. Several techniques are available to grow primary cell cultures from tumors; however, very few have been found to be promising. Novel, cost-effective model that similarly mimic tumor biology and provide faster information on the activity of anticancer therapies could therefore make an important contribution to the advancement of personalized medicine. Chick Chorioallantoic Membrane (CAM) assays have been used to study the process and therapeutics involving angiogenesis, tumor cell invasion and metastasis. The CAM is naturally immuno-deficient and rich in vascularity therefore an ideal system, allowing to generate 3D cancerous “organoids” in a very efficient, reproducible and cost-effective manner and translates basic research to the clinic.

Aim

1) Generate a “personalized medicine” HTP system for a quick and reliable evaluation of the effectiveness of different therapeutic options on of 3D "organoid" tumors; 2) Using the "humanized egg" as a low-cost and highly efficient mimetic of the mouse PDX model ".

Methods

Fertilized eggs were incubated until day 3 (37°C, 75-90% humidity). 2ml of albumin was pulled from the egg to separate the CAM membrane from the egg shell. Then, a small window in the egg shell has been made, and resealed with adhesive tape. On day 7, 3-5x106 tumor cells were transplanted onto the CAM membrane. The cancer cells become a visible tumor after 3 days. Different therapeutics (such as Erbitux, anti-CD24 mAb) were tested. Tumor growth was monitored by "live imaging" device.

Results

The efficiency and reproducibility of human cancer cell lines engraftment has been demonstrated. LV-GFP-Puro plasmid was constructed and used to generate GFP-encoded lentiviruses which then were used to generate a stable colorectal cancer cell line expressing GFP. The CAM tumors were evaluated histopathologicaly and IVIS fluorescent imaging.

Anti-CD24 mAb inhibited tumor growth by ~70%.The presence of the humanized mAb in the engrafted tissue was confirmed by western blot analysis and IHC.

Conclusions

The ability to evaluate ex-vivo cancer tissue response to potential therapeutics in “humanized” settings within days is a very powerful tool.

Citation Format: Shiran Shapira, Oren Bogin, Nadir Arber. The “humanized” chicken embryo chorioallantoic membrane (CAM) as a personalized platform for high-throughput (HTP) screening of cancer therapeutics [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 1168.
Abstract 1180: Quantifying melanoma drug resistance and tumor heterogeneity by live cell interferometry
Authors: Leslie Kevin A. , Huang Dian , Murray Graeme , Guest Daniel , Roy Irena J. , Piva Marco , Moriceau Gatien , Lo Roger S. , Teitell Michael A. , Reed Jason C.
Published: August 2018

Melanoma is the most aggressive type of skin cancer. Tumor heterogeneity and drug resistance are significant obstacles to survival benefits from mutation-targeted therapy. Over 50% of metastatic melanomas harbor V600BRAF mutations. Targeting of MUTBRAF melanoma with BRAF inhibitor monotherapy or the combination of BRAF and MEK inhibitors leads to a high rate of initial responses, but adaptive and acquired resistance frequently leads to clinical relapse. Although the presence of V600BRAF mutations guides the selection of therapy with MAPK inhibitors, methods to track preexisting and adaptive resistance and thereby predict response patterns, or to use this new information to modify therapy, are still lacking. Here, we aimed to develop a rapid, massively parallel tumor cell profiling method, based on Live Cell Interferometry (LCI), to quantify heterogeneous single tumor cell responses and emergent drug resistance. LCI is an ex vivo imaging approach that quantifies changes in total cell biomass, biomass motion, or cell stiffness over time. We report a novel high-throughput screening version of the LCI platform, HSLCI, which rapidly profiles changes in biomass in BRAF inhibitor (BRAFi)-sensitive, parental melanoma cell lines and their isogenic, BRAFi-resistant sub-lines. We show reproducible results from two different HSLCI platforms at two institutions and generate biomass kinetic signatures capable of discriminating between BRAFi-sensitive and -resistant melanoma cells within 24 hours. Our measurements require no fluorescence or dye labeling and are faster than field-standard growth inhibition assays. The accuracy and speed of HSLCI in profiling tumor cell heterogeneity and therapy resistance are promising features of potential tools to guide patient therapeutic selections.

Citation Format: Kevin A. Leslie, Dian Huang, Graeme Murray, Daniel Guest, Irena J. Roy, Marco Piva, Gatien Moriceau, Roger S. Lo, Michael A. Teitell, Jason C. Reed. Quantifying melanoma drug resistance and tumor heterogeneity by live cell interferometry [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 1180.
Abstract 1187: Comparison of intra-tumor heterogeneity and clonal evolution across lung cancer subtypes by multi-region whole genome sequencing
Authors: Zhang Tongwu , Sampson Joshua , Bertazzi Pier Alberto , Pesatori Angela , Consonni Dario , Zhu Bin , Hicks Belynda , Hua Xing , Shi Jianxin , Brown Kevin , Chanock Stephen , Landi Maria Teresa
Published: August 2018

Both lung squamous cell carcinoma (LUSC) and lung cancer with neuroendocrine features (LUNE; i.e., small cell carcinoma (SCLC), large cell carcinoma (LCNEC) and other subtypes) are associated with tobacco smoking and have a very poor prognosis. We sought to characterize their clonal architecture and illustrate how the evolutionary patterns differ across these lung cancer subtypes. We performed multi-region whole genome sequencing of 83 samples from 36 LUSC and 67 samples from 23 LUNE patients that had been resected before systemic therapy.

Among somatic copy number alterations, we observed genome doubling intra-tumor heterogeneity (ITH) and massive genomic instability in both subtypes. For somatic nucleotide variants (SNVs), the total mutation burden was higher in LUSC (16.7 average number of mutations/Mb) than LUNE (12.7/Mb), but for SNVs ITH was higher in LUNE. Among driver genes, TP53, CDKN2A, CR1, and NRK in LUSC and TP53, KRAS, KMT2D, and RB1 in LUNE were frequently mutated and were always clonal. Evidence of positive selection was found for TP53, PTEN, KMT2D and KEAP1 in LUSC, and for TP53 and KRAS in LUNE. As expected, the dominant mutation signatures in LUSC clones included signatures 4, 1, and 5, while APOBEC signatures (2 and 13) were predominantly found in sub-clones. In contrast, no evidence of APOBEC signatures were found in LUNE. Among structural variants, LUNE had more events (339.25 average number of events/tumor) than LUSC (206.7/tumor), with considerable ITH in both subtypes, particularly in LUNE. In LUNE, the dominant mechanism responsible for SVs included fork stalling and template switching within the microhomology-mediated break-induced repair (MMBIR). We found recurrent fusion events with hotspots including those previously identified near TTC28/CHEK2 (42% LUSC and 35% LUNE) and LRP1B (42% LUSC). Additional hotspots were found in chromosome bands 20p12.1 (22% LUSC), 1p31.1 (19% LUSC) and 12p11.22 (22% LUNE). We found >10-fold higher transposon insertions (LINE1) in LUSC (71.9 average number of events/tumor) than LUNE (6.3/tumor). Our findings improve our evolutionary understanding of these lung cancer subtypes and warrant further investigations on the mechanisms involved in clonal genomic instability.

Citation Format: Tongwu Zhang, Joshua Sampson, Pier Alberto Bertazzi, Angela Pesatori, Dario Consonni, Bin Zhu, Belynda Hicks, Xing Hua, Jianxin Shi, Kevin Brown, Stephen Chanock, Maria Teresa Landi. Comparison of intra-tumor heterogeneity and clonal evolution across lung cancer subtypes by multi-region whole genome sequencing [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 1187.
Abstract 121: Angiogenin and the mmp9-timp2 axis are strongly upregulated in pro-angigoenic dnk-like cells isolated from colorectal cancer patients
Authors: Bruno Antonino , Pelosi Giuseppe , Boni Luigi , Dominioni Lorenzo , Mortara Lorenzo , Noonan Douglas , Albini Adriana
Published: August 2018

Natural Killer (NK) cells are effector lymphocytes involved in tumor immunosurveillance. In solid malignancies, tumor associated (TANK; peripheral blood) and tumor infiltrating (TINK) NK cells have compromised functions. We functionally and molecularly characterize TINK and TANKs from blood and tissue samples of colorectal cancer (CRC) patients, a tumor type where inflammation and angiogenesis have clinical relevance, compared to NK isolated from control and non-oncologic inflammatory bowel disease patients.

NK subset distribution and cytokine profiling were performed by multicolor flow cytometry, using peripheral blood and tissue samples from CRC patients, for surface antigen and cytokine profiling characterization. Conditioned media (CM) from FACS-sorted NKs were used either for secretomic profiling using antibody membrane arrays or in functional in vitro angiogenesis assays.

CRC TINKs/TANKs showed decreased activation marker NKG2D, impaired degranulation activity, a decidual-like NK polarization toward the CD56brightCD16dim/- CD9+CD49+ subset. TINKs and TANKs supernatants induced endothelial cell proliferation, migration, adhesion and formation of capillary-like structures in vitro. It has been reported that dNK release proangiogenic factors and metastasis-associated (MMP-9, TIMP1-2) proteins.

We describe for the first time the expression of angiogenin and MMP2, MMP9, TIMP1 and TIMP2 by CRC derived NK cells. This could be a phenotype relevant to their invasive capabilities and pro-angiogenic function. STAT-3/STAT-5 activation was observed in TANKs, and inhibition of the STAT5 pathway by pimozide, an antipsychotic drug, reduced proangiogenic factor VEGF and angiogenin production and capillary-like structure formation, but did not affect the levels of TIMP-1, TIMP-2 and MMP-9.

Citation Format: Antonino Bruno, Giuseppe Pelosi, Luigi Boni, Lorenzo Dominioni, Lorenzo Mortara, Douglas Noonan, Adriana Albini. Angiogenin and the mmp9-timp2 axis are strongly upregulated in pro-angigoenic dnk-like cells isolated from 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 121.
Abstract 1217: The association between birth order and childhood brain tumors: A systematic review and meta-analysis
Authors: Nguyen Mai V. , Tran Mo Thi , Tran Chi TDu , Tran Quang H. , Tuong Thuy Thi-Van , Le Linh C. , Boffetta Paolo , Luu Hung N.
Published: August 2018

Background: The incidence of childhood brain tumors (CBT) has increased worldwide, likely resulting from the improvements of early diagnostics. The only known risk factors for CBT are the Li-Fraumeni syndrome and iodization. While both genetic and environmental factors are suggested roles in CBT, birth characteristics, including birth order, maternal age, and mode of delivery, might present the interactions between genetic susceptibility and perinatal environmental causes. Birth order has been hypothesized to play an important role in CBT carcinogenesis due to its possible role as proxy for 1) early infection exposure and 2) hormone levels since first pregnancy differs endocrinologically from later pregnancies. We conducted a systematic review and meta-analysis to clarify the association between birth order and CBT.

Methods: We followed established guidelines to systematically search OvidMedline, PubMed, and the Cochrane Library for English-language studies (both case-control and cohort designs) published prior to November 2017. Meta-analysis provided pooled risk estimates and their 95% confidence intervals (CI) for birth order and CBT. Funnel plots were used to identify sources of heterogeneity and publication bias.

Results: We identified 16 case-control studies with a total sample of 32,439 cases and 166,144 controls. Compared to first birth order, the meta-odds ratio (mOR) for second birth order was 1.04 (95% CI 1.01-1.07), and that for third birth order was 0.98 (95% CI: 0.90-1.06). We also identified 3 cohorts comprising a total of 1,208 cases. The meta-hazard ratio (mHR) for second or higher birth order compared to first birth order was 1.00 (95% CI: 0.96-1.05).

Conclusions: We found no overall association between birth order and CBT in both case-control and cohort study designs; the small association observed for second birth order in case-control studies deserves further consideration. Limited statistical power might have hampered the results of the other analyses.

Citation Format: Mai V. Nguyen, Mo Thi Tran, Chi TDu Tran, Quang H. Tran, Thuy Thi-Van Tuong, Linh C. Le, Paolo Boffetta, Hung N. Luu. The association between birth order and childhood brain tumors: A systematic review and meta-analysis [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 1217.
Abstract 1219: Association between MICA polymorphisms, s-MICA levels, and pancreatic cancer risk in a population-based case-control study
Authors: Onyeaghala Guillaume C. , Lane John , Pankratz Nathan , Nelson Heather H. , Thyagarajan Bharat , Anderson Kristin E. , Prizment Anna E.
Published: August 2018

Objectives: The immune system has been shown to play an important role in pancreatic cancer. Abnormal cells express the transmembrane major histocompatibility complex class I chain-related gene A (MICA) protein, which is recognized by receptors present on NK and cytotoxic T cells. However, pancreatic tumor cells release MICA protein in soluble form (called s-MICA) from the tumor surface and thus avoid immune surveillance. In our previous study, we showed that a higher serum concentration of the s-MICA protein was associated with increased pancreatic cancer risk. In the current study, we hypothesized that functional variants in the MICA gene are associated with higher circulating s-MICA levels and increased pancreatic cancer risk. We focused on the A5.1 MICA allele, as this allele encodes a MICA protein that is shorter than its normal counterpart and is more easily cleaved from the cell surface.

Methods: MICA alleles and s-MICA levels were measured in 116 cases and 492 controls (n=608) in a population-based case-control study in Minnesota. Allele assignments were based on the number of repeat units in the amplified regions of the MICA gene, and s-MICA levels were measured by enzyme-linked immunoabsorbent assay. To address the non-normal distribution of s-MICA levels, general linear regression with a log link was used to assess mean s-MICA levels across MICA alleles. Unconditional logistic regression was used to calculate the odds ratio (OR) and 95% confidence intervals (CI) for pancreatic cancer after adjustment for age, sex, education, smoking status, alcohol consumption, and diabetes status. The analysis was restricted to Caucasians, who represented 96% of all study participants.

Results: 452 participants (74%) possessed at least one copy of the A5.1 allele. After adjustment for confounders, participants with at least one copy of the MICA A5.1 allele were at an increased risk of pancreatic cancer (multivariable OR=2.00, 95% CI: 1.06-3.79), compared to participants who did not possess any A5.1 allele. Participants who had at least one copy of the A5.1 allele had 2.46 (95%CI: 2.36-2.56) times greater mean s-MICA levels than those without the A5.1 allele. These associations were consistent among pancreatic cancer cases and controls.

Conclusions: Our study supports the role of the MICA A5.1 allele in impaired immune response, which may increase the risk of pancreatic cancer.

Citation Format: Guillaume C. Onyeaghala, John Lane, Nathan Pankratz, Heather H. Nelson, Bharat Thyagarajan, Kristin E. Anderson, Anna E. Prizment. Association between MICA polymorphisms, s-MICA levels, and pancreatic cancer risk in a population-based case-control study [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 1219.
Abstract 1225: Rare variants in DNA damage repair genes are associated with male breast cancer predisposition
Authors: Maguire Sarah L. , Tomczyk Katarzyna , Perrakis Eleni , Saunders Edward , Leongamornlert Daniel , Kote-Jarai ZSofia , Eeles Rosalind , Garcia-Closas Montserrat , Pharoah Paul , Easton Douglas , Lord Christopher J. , Ashworth Alan , Swerdlow Anthony , Orr Nick
Published: August 2018

Male breast cancer accounts for approximately 1% of all breast cancers diagnosed each year in the UK. Germline mutations in DNA damage response genes are known to be associated with many forms of cancer, including breast cancer. In order to identify low frequency male breast cancer predisposition genes, we performed targeted resequencing of more than 300 genes that were selected on the basis of involvement in DNA maintenance and repair pathways. Using a cost-effective protocol based on an automated library preparation method and a multiplexed hybridization enrichment step, we sequenced 1,029 male breast cancer cases from the UK Breast Cancer Now male breast cancer study (MBCS) and 1,254 controls from the MBCS and the ICR1000 UK exome series. Rare variants localising to 19 genes were significantly associated with risk of male breast cancer. Genes with an excess of rare coding mutations included the known breast cancer susceptibility genes BRCA2, CHEK2 and ATM as well as multiple members of the Fanconi anaemia (FA) pathway. Loss of heterozygosity (LOH) affecting the wildtype allele of putative predisposition genes was observed more than 10% of cases with matched tumor samples. In addition to LOH in BRCA2 mutation carries, somatic losses were observed that affected CCNE1, ATM, PALB2, BARD1, CHEK2, POLL and RAD9A. These data represent the largest analysis of rare variants in male breast cancer to date and have enabled the identification of known and putative novel susceptibility genes.

Citation Format: Sarah L. Maguire, Katarzyna Tomczyk, Eleni Perrakis, Edward Saunders, Daniel Leongamornlert, ZSofia Kote-Jarai, Rosalind Eeles, Montserrat Garcia-Closas, Paul Pharoah, Douglas Easton, Christopher J. Lord, Alan Ashworth, Anthony Swerdlow, Nick Orr. Rare variants in DNA damage repair genes are associated with male breast cancer predisposition [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 1225.
Abstract 1272: Docosahexaenoic acid supplementation enhances the response of PC-3 prostate cancer cells to enzalutamide
Authors: Ma Irvin V. , Cooper Andrew R. , Hu Ying , Sorreta Arianne G. , Bains Palvinder K. , Kumar Prem , Pardini Ronald S.
Published: August 2018

Prostate cancer is currently the second leading cause of death from cancer among men in the United States. Initial treatments generally focus on hormone therapy to inhibit the production of androgens, but most prostate cancers eventually become castration resistant and no longer respond to androgen-suppression therapy. The recent development of drugs that inhibit androgen binding to the androgen receptor (AR) have proven effective in treating castration-resistant prostate cancer (CRPC). Studies in our laboratory have shown that docosahexaenoic acid (DHA; C22:6 n-3), a long chain omega-3 polyunsaturated fatty acid (PUFA), inhibits tumorigenesis in both androgen-dependent (LNCaP) and androgen-independent (PC-3, DU145) prostate cancer cell lines in culture. Moreover, in vivo studies with n-3 PUFA supplementation have been shown to significantly reduce tumor growth of PC-3 and DU145 compared to corn oil diets rich in linoleic acid, (C18:2, n-6). With evidence suggesting that n-3 PUFA dietary supplementation can inhibit tumorigenesis in CRPC, the current study investigated the impact of DHA supplementation in combination with enzalutamide in PC-3 tumor cells. Enzalutamide is a Food and Drug Administration-approved treatment for patients with metastatic CRPC. Enzalutamide acts by interrupting the binding of androgens to the AR, which influences the AR signaling pathway–a target of interest in CRPC treatment. Cell viability was determined by measuring intracellular ATP in PC-3 cells treated with DHA and enzalutamide alone and in combination. Cell viability was reduced with the combination of DHA and enzalutamide compared to enzalutamide or DHA alone, suggesting that DHA enrichment can augment enzalutamide therapy with PC-3 cells in vitro. Studies have also shown that combinatorial treatments with enzalutamide and HIF-1α inhibitors result in synergistic inhibition of CRPC tumor proliferation, and our laboratory has shown that DHA inhibits HIF-1α by decreasing intracellular ATP in breast and lung cancer cells in vitro. These results serve as a prelude to a pilot clinical trial to evaluate the effects of nutritional supplementation with high levels of omega-3 fatty acids on enzalutamide inhibition of CRPC.

Citation Format: Irvin V. Ma, Andrew R. Cooper, Ying Hu, Arianne G. Sorreta, Palvinder K. Bains, Prem Kumar, Ronald S. Pardini. Docosahexaenoic acid supplementation enhances the response of PC-3 prostate cancer cells to enzalutamide [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 1272.
Abstract 1273: Effect of a Western diet on colitis-associated colon tumor formation in BALB/c mice
Authors: Groeschel Charlotte , Tennakoon Samawansha , Aggarwal Abhishek , Kallay Enikoe
Published: August 2018

Patients with inflammatory bowel disease (IBD) are at increased risk for inflammation associated colorectal cancer (CAC). The incidence of IBD is increasing in the Western industrialized world. Epidemiological studies suggest that a diet, high in fat but low in vitamin D, calcium, fibre and methyl donors is a risk factor for both, IBD and colorectal cancer (CRC). It has been shown previously that a ”New Western Diet”, caused development of sporadic colonic tumours in mice.

Our study was performed to evaluate whether a Western Diet (WD) aggravates azoxymethane (AOM) /dextran sodium sulfate salt (DSS)-induced colitis-associated carcinogenesis in mice, and if a switch to the standard purified mouse diet AIN93G after cancer initiation is still able to ameliorate or delay the onset of the disease. Female BALB/c mice received either the WD (0.5 mg/g calcium, 2% fibre, 21 % butter fat, 0.23 μg/g folic acid) or AIN93G standard diet for 5 weeks until AOM was administered at a dose of 12.5 μg/kg intraperitoneally, followed by 3 cycles of DSS (2.5%). In one group the WD was switched to AIN93G diet one day before first administration of DSS. The mice were euthanized 80 days after AOM injection. One part of the colon, small intestine, kidney and liver were frozen in liquid nitrogen, the rest formalin fixed and paraffin embedded for mRNA and immunhistochemical analysis. Feeding constantly the WD shortened the colon (p<0.05) and increased the number (p<0.05) and size (p<0.01) of aberrant colonic crypt foci. The switch to the AIN93G diet ameliorated this effect, leading to reduced colitis and tumor promotion comparable with the AIN93G group. In colon ascendens of the WD group, Ki67 protein levels (p<0.001) and relative mRNA levels of the Wnt target genes c-myc (p<0.05) and axin2 (p<0.001) were reduced while mRNA levels of the vitamin D catabolizing enzyme cyp24a1 (p<0.001) and inos (p<0.05) were increased when compared with the AIN93G group.

A healthy diet protects colonic mucosa against AOM/DSS-treatment and is an effective chemopreventive strategy to reduce chronic colonic inflammation and tumorigenesis.

This project is funded by the Vienna Science and Technology Fund and the Austrian Research Fund.

Citation Format: Charlotte Groeschel, Samawansha Tennakoon, Abhishek Aggarwal, Enikoe Kallay. Effect of a Western diet on colitis-associated colon tumor formation in BALB/c mice [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 1273.
Abstract 1288: A comprehensive investigation of factors impacting the accuracy of mutation detection using next generation sequencing technology
Authors: Xiao Wenming , Zhao Yongmei ,   Somatic Mutation Working Group, SEQC2 Consortium
Published: August 2018

Next generation sequencing (NGS) allows researchers to survey DNA mutations in tumor cells at very large scales with affordable cost. However, distinguishing tumor cell sub-population mutations from errors due to NGS platform biases, sample preservation effects, assay protocols and suboptimal or incongruous bioinformatics analysis methods is currently complex and difficult. For NGS to be successfully leveraged in a clinical translational manner, increased ability to discriminate between true mutations and artifacts is paramount.

To address this urgent unmet clinical need, we systematically investigated somatic mutation events in paired cell lines (breast cancer and normal) with multiple NGS platforms and protocols to fully characterize factors that affect accuracy, specificity and sensitivity of mutation detection. We applied machine learning algorithms to define high confidence mutation calls with data sets from 8 NGS platforms combined with 9 bioinformatic analysis pipelines. With “ground truth” established using multiple orthogonal sequencing platforms, we evaluated the performance of mutation detection under various circumstances, including: fresh cell vs FFPE, amount of DNA, library preparation, sequencing platform, tumor purity, and analysis algorithm. Moreover, we performed whole exome sequencing (WES) and whole genome sequencing (WGS) at 6 sequencing sites in parallel, which allowed us to assess the reproducibility of NGS runs on the same biological samples.

By analyzing DNA variant detection recall and precision from data sets derived from various conditions, we identified statistical variance components in overall DNA variant detection framework as well as specific practices that will likely lead to the false identification of variants. By concomitantly examining the effects between sample processing and bioinformatics pipelines, here we recommend best practices for mutation detection using NGS technology and establish quality metrics for cancer studies involving NGS technology.

Citation Format: Wenming Xiao, Yongmei Zhao, Somatic Mutation Working Group, SEQC2 Consortium. A comprehensive investigation of factors impacting the accuracy of mutation detection using next generation sequencing technology [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 1288.
Abstract 13: Anti-angiogenic action of leukotriene-C4induced 15-hydroxyprostaglandin dehydrogenase in colon cancer cells is a TNF-α dependent phenomenon
Authors: Satapathy Shakti R. , Sjölander Anita
Published: August 2018

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. Cyclooxygenase-2 (COX-2), which plays a key role in the biosynthesis of prostaglandin E2 (PGE2), is often up-regulated in CRC and in other types of cancer. PGE2 induces angiogenesis and tumor cell survival, proliferation and migration. The tumor suppressor 15-hydroxyprostaglandin dehydrogenase (15-PGDH) is a key enzyme in PGE2 catabolism, converting it into its inactive metabolite 15-keto-PGE2 and is often down-regulated in cancer. Interesting enough, CRC patients expressing high levels of Cysteinyl leukotriene receptor 2 (CysLTR2) have a good prognosis and therefore, we investigated a potential link between CysLTR2-signaling and the tumor suppressor 15-PGDH in colon cancer cells. TNF-α is considered as the main regulators of COX-2 and mPGES-1 that contribute to the increased synthesis of PGE2, which is inhibited by overexpressed 15-PGDH. Level of the pro-tumorigenic PGE2 are increased in CRC, previously attributed to increased production via TNF-α mediated COX-2 up-regulation but more recently attributed to decreased catabolism due to down-regulation of 15-PGDH. Elevation of 15-PGDH expression by leukotriene C4 (LTC4), a CysLTR2 ligand, exhibited anti-tumor activity in colon cancer cells with significant phosphorylation of β-catenin and down-regulation of anti-apoptotic marker Bcl-2 with concurrent activation of CASPASE-3 expression. We also observed a dramatic down-regulation of TNF-α on mRNA level and NF-κβ on both mRNA as well as protein level with LTC4 induced 15-PGDH in a TNF-α dependent manner. Moreover, TNF-α regulated anti-angiogenic action of 15-PGDH in HT-29 and Caco-2 colon cancer cells by depleting the mRNA level of MMP-2 and MMP-9 and protein level of VEGFR-1. Furthermore, our preliminary observation also suggested disrupted tube formation in HUVEC with LTC4 induced 15-PGDH which is mediated by TNF-α. Hence, restoration of 15-PGDH expression through CysLTR2-signaling promotes the anti-angiogenic action against colon cancer cells, indicating an anti-tumor as well as the anti-metastatic efficacy of CysLTR2-signaling.

Citation Format: Shakti R. Satapathy, Anita Sjölander. Anti-angiogenic action of leukotriene-C4induced 15-hydroxyprostaglandin dehydrogenase in colon cancer cells is a TNF-α dependent phenomenon [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 13.
Abstract 1310: Systematic evaluation of gene networks for discovery of disease genes
Authors: Huang Justin K. , Carlin Daniel E. , Yu Michael K. , Zhang Wei , Kreisberg Jason F. , Tamayo Pablo , Ideker Trey
Published: August 2018

Gene networks are rapidly growing in size and number, raising the question of which networks are most appropriate for a particular application. Here, we evaluate 21 human genome-wide interaction networks for their ability to recover gene sets associated with 446 different diseases and 9 cancer hallmarks. While all networks have some ability in these recovery tasks, we observe a wide range of performance with STRING, GeneMANIA and GIANT networks having the best performance overall. A general tendency is that performance scales with network size, suggesting that new interaction discovery currently outweighs the detrimental effects of false positives. Correcting for size, we find that the DIP network provides the highest efficiency (value per interaction). Based on these results we create a parsimonious composite network with both high efficiency and absolute performance, which outperforms any single resource. This work provides a benchmark for selection of molecular networks in human disease research.

Citation Format: Justin K. Huang, Daniel E. Carlin, Michael K. Yu, Wei Zhang, Jason F. Kreisberg, Pablo Tamayo, Trey Ideker. Systematic evaluation of gene networks for discovery of disease genes [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 1310.
Abstract 1323: A p53-dependent switch between cytoprotective and nonprotective autophagy influences tumor cell sensitivity to cisplatin
Authors: Xu Jingwen , Saleh Tareq , Patel Nipa , Wu Yingliang , Lima Santiago , Gewirtz David
Published: August 2018

Autophagy is a multi-step, homeostatic process through which cells turnover dysfunctional organelles and proteins. Previous work in our laboratory has identified multiple functional roles of autophagy, specifically cytoprotective, cytotoxic, cytostatic and nonprotective, when subjected to radiation and chemotherapy. Furthermore, we have reported on the existence of an autophagic switch between one functional form (generally cytoprotective) to the other forms. To further explore the autophagic switch, we utilized p53 wild type (p53 +/+) and p53 null (p53 -/-) H460 non-small cell lung cancer (NSCLC) cell lines exposed to cisplatin. The extent of autophagy was similar in both cell lines, but the rate and extent of apoptosis was higher in the p53 +/+ cells, which showed greater cisplatin sensitivity. Pharmacological (chloroquine, 3-Methyladenine) and genetic (ATG5 silencing) inhibition of autophagy failed to sensitize the p53 +/+ H460 cells to cisplatin, suggesting that autophagy has a nonprotective role. In contrast, autophagy inhibition (CQ, 3-MA, or shATG5) increased sensitivity to cisplatin in the p53 -/- H460 cells, indicative of a cytoprotective function of autophagy. These studies support the premise that cytoprotective autophagy may confer resistance to cisplatin. Moreover, we confirm the pivotal role of p53 in determining the function of autophagy induced by cisplatin. Finally, we provide an additional example of a switch between the nonprotective and protective forms of autophagy in p53 wild type and p53 null H460 non-small cell lung cancer (NSCLC) cells, respectively.

Citation Format: Jingwen Xu, Tareq Saleh, Nipa Patel, Yingliang Wu, Santiago Lima, David Gewirtz. A p53-dependent switch between cytoprotective and nonprotective autophagy influences tumor cell sensitivity to cisplatin [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 1323.