bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5278: Separate and combined effects of metabolic reprogramming interventions, autophagy inhibition, and carboplatin on murine triple-negative breast cancer cells
Authors: Cozzo Alyssa J. , Coleman Michael F. , O'Flanagan Ciara H. , Pearce Jane B. , Rainey Magdalena A. , Hursting Stephen D.
Published: August 2019

Triple-negative breast cancers (TNBC) are an aggressive BC subset currently lacking targeted therapies. TNBCs exhibit aberrant fuel metabolism, including increased flux through glycolysis and the pentose phosphate pathway, increased lactate production (Warburg metabolism), and elevated dependence on glutamine (glutaminolysis). While dietary interventions such as caloric restriction, intermittent fasting, or ketogenic diets have demonstrated benefits (e.g, reduced tumor incidence, progression, and sensitization to some chemotherapeutic agents) in preclinical BC models, translation to humans has proven challenging, and concerns remain regarding tolerance and long-term safety of extreme diets in cancer patients. We hypothesized that targeted disruption of select metabolic pathways could recapitulate the beneficial effects of the aforementioned dietary interventions and improve the efficacy of carboplatin, part of first-line chemotherapy for primary and metastatic TNBCs. Moreover, we posited that blocking autophagy would further enhance the anticancer effects of metabolic reprogramming interventions (MRIs) alone and in combination with carboplatin. To test these hypotheses, we are using a high-throughput screening approach employing several MRIs with or without the addition of chloroquine and/or carboplatin in vitro. Serum starvation (an in vitro model of caloric restriction), rapamycin, BMS-754807 (a dual IGF-1R/insulin receptor), and nicotinamide significantly reduced cell viability (assessed by MTT assay) and clonogenicity (assessed by colony formation assay) of M-Wnt and M6 cells, both well-characterized TNBC cell lines derived from mammary tumors of MMTV-Wnt-1 and C3-TAg transgenic mice, respectively. The growth suppressive effects of these MRIs were further enhanced when combined with autophagy inhibition via chloroquine (CQ). Treatment of M-Wnt and M6 cells with CQ (20-80 µM) also increased in vitro cytotoxicity of carboplatin. Ongoing analyses and planned in vivo studies will confirm whether co-treatment with MRIs and/or CQ are an effective strategy for increasing the efficacy of carboplatin in TNBC cells.

Citation Format: Alyssa J. Cozzo, Michael F. Coleman, Ciara H. O'Flanagan, Jane B. Pearce, Magdalena A. Rainey, Stephen D. Hursting. Separate and combined effects of metabolic reprogramming interventions, autophagy inhibition, and carboplatin on murine triple-negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5278.
Abstract 5280: Branched chain amino acids represent indispensible metabolic substrates in glioblastoma
Authors: Prabhu Antony H. , Kant Shiva , Kesarwani Pravin , Chinnaiyan Prakash
Published: August 2019

Glioblastoma (GBM) is an aggressive brain tumor with limited treatment options. Considerable advances have been made in understanding the genetic alterations driving these tumors, however, the metabolic alterations and mechanisms underlying these aggressive phenotypes remain unclear. We performed global metabolomic profiling of patient derived low-grade astrocytoma (LGA, n= 28) and GBM (n=80). Hierarchical clustering (HC) identified amino acid (AA) metabolism as a dominant metabolic node in GBM compared to LGA. Considerable inter-tumoral heterogeneity of AA metabolism was observed in GBM, with HC identifying two clearly distinct metabolic subtypes consisting of increased and decreased concentrations of AA and their metabolic intermediates. VIP analysis identified the accumulation of branched chain amino acids (BCAA)- leucine, valine and isoleucine as the top ranked metabolites differentiating these subtypes. Based on this, GBMs were classified as BCAA-high and BCAA-low. BCAA-high GBM was enriched with mesenchymal and classical subtypes while BCAA-low GBM was enriched with proneural and neural subtypes. Integrative analysis coupling these metabolic signatures with gene expression profiling performed on matched tissue demonstrated upregulation of BCAA metabolism-associated genes in BCAA-high GBM, along with global transcriptional programs designed to activate cell cycle progression. When extended into molecular subtype specific preclinical models, we demonstrated that BCAA were indispensible for survival of mesenchymal GBM cells, which was independent of the presence or absence of other non-essential and essential amino acids. Further, these cells were able to maintain long-term survival in a dormant state when grown in media devoid of all essential and non-essential AA other than BCAA and glutamine. Knockdown of key enzymes in BCAA metabolism led to robust cytotoxicity in this model, underscoring the importance of BCAA in GBM survival. Ongoing studies are designed to determine the biologic consequence and intermediary metabolism of BCAA in GBM. Collectively these results identify BCAA metabolism as a therapeutic target in GBM.

Citation Format: Antony H. Prabhu, Shiva Kant, Pravin Kesarwani, Prakash Chinnaiyan. Branched chain amino acids represent indispensible metabolic substrates in glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5280.
Abstract 5294: The PanNET-related histone H3.3 chaperone Daxx regulates lineage specification and tissue homeostasis in the pancreas
Authors: Sposito Teresa , Tu Marie-Sophie Nguyen , Rutter Guy A. , Novelli Marco , Thirlwell Chrissie , Salomoni Paolo
Published: August 2019

Progressive age-dependent replacement of canonical H3.1/2 with H3.3 in post-mitotic cells is a crucial mechanism for maintaining genome integrity at heterochromatic sites. The histone chaperone DAXX cooperates with the chromatin remodeller ATRX for the deposition of the non-canonical histone H3.3 at heterochromatin, while it prevents the expression of endogenous retroviral elements in another complex involving the SETDB1 methyltransferases. Loss of function mutations affecting ATRX/DAXX and MEN-1 have been recently identified as a subgroup of pancreatic neuroendocrine tumours (PanNETs) leading to poor prognosis and more aggressive phenotype compared to the ATRX/DAXX and MEN-1 wild-type counterparts. We developed a mouse model for conditional Daxx inactivation in mouse islets to understand the impact of DAXX loss on pancreas homeostasis and PanNETs development. We found that loss of Daxx in the endocrine pancreas, using the Insulin-1Cre driver, leads to aberrant glucose metabolism with the development of hyperglycemia and consequently insulin resistance in mice. We have also shown that loss of DAXX is linked to increased expression of α-cell lineage genes upon glucose stimulation of isolated islets in vitro, resembling the recently reported phenotype of PanNETs harboring ATRX/DAXX or MEN1 mutations. Nonetheless, loss of Daxx was not sufficient to lead to PanNET development. We are currently investigating the chromatin and transcriptional perturbations caused by Daxx loss and how these could be linked to the aberration of cell lineage specification and tissue dysfunction in the pancreas. More generally, this work will provide new insight into how epigenetic alterations contribute to misregulation of developmental programmes and cancer development in the pancreas.

Citation Format: Teresa Sposito, Marie-Sophie Nguyen Tu, Guy A. Rutter, Marco Novelli, Chrissie Thirlwell, Paolo Salomoni. The PanNET-related histone H3.3 chaperone Daxx regulates lineage specification and tissue homeostasis in the pancreas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5294.
Abstract 5298: Finding new molecules to bypass the tumor suppressor gene TP53 and restore cell cycle checkpoint control
Authors: Moore Henry , Blackburn Jessica
Published: August 2019

The tumor suppressor gene, tumor protein 53 (TP53), has been shown to be mutated in more than half of all human cancer cases. p53 controls the G1/S phase checkpoint of the mitotic cell cycle, protecting the fidelity of DNA and inducing apoptosis in cells with irreparable DNA damage. Mutations in TP53, which are often missense mutations that interfere with p53 DNA binding activity or protein folding, result in protein inactivation and loss of checkpoint control, allowing cells with potentially oncogenic mutations to survive and proliferate. TP53 mutations have been shown to enhance tumor progression, increase metastatic potential, and mediate chemotherapy resistance. Because of the importance of p53 in cancer, many studies have aimed to identify small molecules that can “reactivate” mutated p53 protein in cell culture, although this has met with little success thus far. The aim of this research is to identify the extent to which the effects of non-functioning p53 can be completely bypassed, and cell cycle checkpoint restored, using already-established FDA approved compounds in a zebrafish model of p53 loss.

Zebrafish were genetically engineered to have their entire TP53 locus removed using TALEN (Transcription activator-like effector nuclease), rendering them unable to produce the p53 protein. p53 deficient and wild-type zebrafish larvae were then subjected to gamma-irradiation to induce DNA damage. An apoptotic cell staining assay was then performed to allow for the qualitative and quantitative analysis of apoptosis levels throughout the larvae. As expected, TP53 null fish had significantly less apoptotic cells than control after DNA damage. We are in the process of testing 770 FDA approved compounds for their effects in inducing an apoptotic response in p53 null larvae after gamma-irradiation. Positive hits will be carried forward to human cancer cells with TP53 mutation, and the mechanism of action of the drug will be determined using biochemical approaches. These results could identify novel pathways that work in concert with p53 and will allow us to harness new mechanisms of DNA damage induced apoptosis to kill cancer cells. Repurposing FDA-approved drugs can also rapidly translate to clinical benefit in a wide number of cancers driven by TP53 mutation.

Citation Format: Henry Moore, Jessica Blackburn. Finding new molecules to bypass the tumor suppressor gene TP53 and restore cell cycle checkpoint control [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5298.
Abstract 530: Phosphatidylserine-targeting antibody combined with chemoradiation enhances systemtic anti-tumor immunity
Authors: Wang Yifan , Liu Zhigang , Deng Weiye , Kim Betty Y. , Lin Steven H. , Jiang Wen
Published: August 2019

The discovery of immune checkpoint blockades have resulted in significant clinical benefits for patients with metastatic cancers including non-small cell lung cancer (NSCLC). Nevertheless, a large proportion of patients do not respond to such therapy. Strategies to enhance the therapeutic efficacy of cancer immunotherapy among a larger group of patients are urgently needed. Increasing the recognition of cancer cells by immune system may enhance response to immunotherapies. Tumor-associated macrophage (TAM) is the most abundant immune cell in most solid tumors, and is critical in bridging innate and adaptive immune system functions. Cancer cells overexpress multiple immune suppressive molecules including phosphatidylserine (PS) to inhibit immunological functions of TAMs. PS expression can be enhanced by cancer therapeutics such as radiation to induce immune suppression. Our results show that radiation increased expression of PS on lung cancer cells. The combination of PS blocking antibody with radiation decreased immune-suppressive phagocytosis and enhanced antigen presentation of macrophages. In murine lung cancer models, combination treatment of PS blockade and chemoradiation more effectively controlled tumor growth and prolonged survival than each treatment used alone. In a bilateral lung cancer tumor model, addition of PS blockade to chemoradiation promoted cytotoxic T cell infiltration and induced tumor growth inhibition of the non-irradiated tumors. Together, our results demonstrate that PS-targeting antibodies may be combined with chemoradiation to enhance intrinsic tumor immunogenicity and activate systemic anti-tumor immune responses. This approach may provide new routes for improving the efficacy of modern immunotherapeutics.

Citation Format: Yifan Wang, Zhigang Liu, Weiye Deng, Betty Y. Kim, Steven H. Lin, Wen Jiang. Phosphatidylserine-targeting antibody combined with chemoradiation enhances systemtic anti-tumor immunity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 530.
Abstract 5300: LIN9 as a potential negative regulator of the glioblastoma stem cell state
Authors: Farkouh Farid , Chheda Milan G. , Jash Arijita
Published: August 2019

Introduction: Glioblastoma (GBM) is the most lethal primary brain tumor. The GBM stem cells are treatment-resistant cells. We performed an RNAi screen to identify genetic drivers of the GSC state. LIN9 suppression scored strongly in the opposite direction: LIN9 suppression led to a significant increase in neurosphere size and number. In other cellular contexts, LIN9 interacts with pRB and cooperates to promote differentiation. It also inhibits DNA synthesis and oncogenic transformation. The purpose of this study is to better understand the role of LIN9 in GBM.

Methods: Lentivirus for suppression and overexpression of LIN9 was produced via transfection of 293T cells. We transduced patient-derived 0308 GSCs. qRT-PCR was used to determine the expression of various genes.

Results: LIN9 suppression in 0308 GSCs results in increased sphere formation and increased cell proliferation but not increased cell size. Additionally, LIN9 suppression results in upregulation of Sox2 in human astrocytes. In contrast, LIN9 overexpression in 0308 cells results in decreased sphere formation and decreased proliferation. Further ongoing experiments will determine if suppression and overexpression of LIN9 have an effect on the expression of stem cell markers (ex. SOX2, Nestin, OLIG2) and differentiation markers (ex. GFAP, MAPK2, GAL4). We expect LIN9 suppression will result in an increase in the expression of stem cell markers and a decrease in expression of differentiation markers. We also expect the opposite results for LIN9 overexpression.

Conclusions: LIN9 is a potentially important negative regulator of the glioblastoma stem cell state.

Citation Format: Farid Farkouh, Milan G. Chheda, Arijita Jash. LIN9 as a potential negative regulator of the glioblastoma stem cell state [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5300.
Abstract 533: The immune landscape in colon cancer and its changes after radiotherapy
Authors: Nigro Cristiana Lo , Sangiolo Dario , Ricci Vincenzo , Garrone Ornella , Abbona Andrea , Falletta Antonella , Merlotti Anna , Varamo Chiara , Aglietta Massimo , Gammaitoni Loretta , Bertetto Oscar , Merlano Marco
Published: August 2019

Background: The TRANSLATE project started in 2016 to test the immune effects on circulating immune cells and cytokines of metronomic Cyclophosphamide (CTX), daily low-dose IL-2 administered every other week, and radiotherapy (RT). The rationale is based on self vaccination induced by RT, T cells expansion by IL-2 treatment and selective Tregs down regulation by low dose cyclophosphamide administration.

Materials/Methods: Treatment consisted of CTX 50/mg tab daily continuously until progression or unacceptable toxicity, 8 Gy single fraction on one metastatic lesion given day 8 from CTX start and IL-2, 500.000 UI twice a day i.v. day 1 to 5 every other week, starting the day after RT. We enrolled patients with end-stage for colon, breast, kidney and prostate cancer. Analysis was performed at baseline, the day after RT, after 28 days from treatment start and at disease progression. Assay focused on Tregs, CD8+, NK, MDSC, CD3-PD1, IL-2, IL-4, IL-5, IL- 6, IL-10, IL-12, IL-13, IL-17a, TNFα, IFNγ, TGFβ. We report baseline and post-RT data separately in patients with colon cancer (CC) and other patients (non-CC).

Results: We report data from 8 pts with left colon, 6 with breast, 2 with kidney and 3 with prostate cancer. At baseline, cytokine expression was similar in CC and non-CC except IFNγ and IL-13, both less represented in CC. After RT, TNFα increased in both groups, but more evidently in CC. A clear increase in IL-10 and IFNγ occurred only in non-CC, while IL-2 increased only in CC. Cumulatively, CC and non-CC reacted differently to RT in terms of positive or negative changes of the analyzed cytokines. On the contrary, we did not observe clear differences in immune cells between CC and non-CC neither at baseline nor after RT.

Discussion: The limited number of patients reduced the interpretation of the study. The differences observed are numerical, but not statistically significant. Colon cancer shows some peculiarities, including the low IL-13 and IFNγ value and a different response to RT. Further analyses investigations are in progress.

Citation Format: Cristiana Lo Nigro, Dario Sangiolo, Vincenzo Ricci, Ornella Garrone, Andrea Abbona, Antonella Falletta, Anna Merlotti, Chiara Varamo, Massimo Aglietta, Loretta Gammaitoni, Oscar Bertetto, Marco Merlano. The immune landscape in colon cancer and its changes after radiotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 533.
Abstract 563: A biparatopic agonist antibody for OX40 that exhibits superior activity without secondary crosslinking
Authors: Hammer Bonnie , Bawadekar Mandar , Bissen Matthew , Painter John , Lehmann Lauren , Li Francis Qufei , Bailey Lucas , Glaser Bryan , Green Roland
Published: August 2019

The development of agonistic antibodies that activate T-cell co-stimulatory pathways represents a therapeutic strategy with significant clinical potential. However, challenges remain for the translation from in vitro efficacy to clinical success. OX40 and other tumor necrosis factor receptor (TNFR) superfamily members are notorious for requiring high-order receptor clustering in order to achieve full activity. For monoclonal antibodies, this high-order clustering is generally achieved through secondary cross-linking strategies. In vivo, this secondary cross-linking is often supplied through internal immune effector cells via Fc engagement. Bispecific and biparatopic antibodies represent an emerging class of drug molecules that enable unique mechanisms of action relative to their monoclonal counterparts. Here, we describe the use of our bispecific platform for the generation of large panels of biparatopic antibodies which enabled high-throughput screening for the discovery of an array of OX40 agonistic molecules. We have optimized these multivalent antibodies that exceed the potency of the OX40 ligand in NFkB activation without the need for secondary cross-linking. These agonist antibodies have additionally been characterized using primary T cell assays to monitor the kinetics of growth proliferation and cytokine secretion, outperforming cross-linked antibodies currently being tested in clinical trials. In co-culture systems, these agonist antibodies were effective in inhibiting immuno-suppressive properties of Tregs. Our lead OX40 agonist antibody has been optimized for activity and developability and has entered stable cell line development to further support pre-clinical activities.

Citation Format: Bonnie Hammer, Mandar Bawadekar, Matthew Bissen, John Painter, Lauren Lehmann, Francis Qufei Li, Lucas Bailey, Bryan Glaser, Roland Green. A biparatopic agonist antibody for OX40 that exhibits superior activity without secondary crosslinking [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 563.
Abstract 577: Optimization of methods for the analysis of class I MHC peptides by mass spectrometry
Authors: Ford Michael J. , Jones Richard C. , Amunugama Ravi , Allen David , Rizzo Paul Del , Mobley James , Pisano Michael
Published: August 2019

Immuno-oncology describes therapeutic approaches exploiting the body's immune system to fight cancer. One approach involves educating the immune-system to recognize and destroy tumor cells by targeting tumor-specific neoantigens. Neoantigens are antigens presented by tumors but not recognized by the immune-system. Identification of Neoantigens is therefore an active area of research and development. The major histocompatibility complex (MHC) plays a crucial role in antigen presentation. Peptides generated by protein degradation in the cytosol are presented, non-covalently bound to MHC Class I molecules, on the surface of cells for inspection by T-lymphocytes. Cytotoxic T lymphocytes (CTL) recognize peptides presented by MHC Class I. The recognition of peptide antigens presented by MHC Class I results in the destruction of the presenting cell by the CTL. Characterization of peptides associated with MHC Class I molecules requires a targeted protein complex enrichment, an unbiased peptide elution and finally a peptide analysis method. Here we present the latest results from our work optimizing and performing a workflow for the analysis of peptides associated with Class I MHC molecules.

Citation Format: Michael J. Ford, Richard C. Jones, Ravi Amunugama, David Allen, Paul Del Rizzo, James Mobley, Michael Pisano. Optimization of methods for the analysis of class I MHC peptides by mass spectrometry [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 577.
Abstract 580: Detailed analysis of immune responses induced against patient-specific neoantigens using the ex-vivo induction protocol, NEO-STIM™
Authors: Lenkala Divya , Kohler Jessica , McCarthy Brian , Nelson Michael , DeBarge Rachel , Ware Yvonne , Huang Yuting , Sridar Janani , Nasrullah Yusuf , Hartjano Dewi , Poran Asaf , Sengupta Sejuti , Berg Joost H. van den , Goldstein Matt , Gaynor Richard B. , Buuren Marit M. van
Published: August 2019

Background: Neoantigens are tumor-specific antigens that have been shown to be critical in the anti-tumor immune response. These antigens are not subject to central immune tolerance and are therefore potentially more immunogenic than tumor-associated antigens. The goal of our studies is to assess the immunogenic potential of high-quality neoantigen targets, as defined through our bioinformatic engine RECON®, as well as perform detailed characterization of the induced T cell responses towards these neoantigen targets.

Methods: Patient-specific neoantigens were predicted using our bioinformatic engine RECON. The predicted high quality neoantigens were utilized in our proprietary ex-vivo stimulation protocol NEO-STIM to assess immunogenicity. NEO-STIM is used to prime, activate and expand memory and de novo T cell responses from both the CD4+ as well as the CD8+ compartment. In-depth analysis was performed to characterize the specificity, functionality (cytokine production and cytolytic capacity) and diversity of the induced T cell responses.

Results: Here we present the successful induction of memory and de novo CD8+ T cell responses in PBMCs isolated by leukapheresis from three patients using NEO-STIM (2 with melanoma & 1 with NSCLC). We then deeply characterize the induced CD8+ T cell responses of one of the melanoma patients and show that these responses are functional, specific and have cytolytic capacity. Furthermore, we use paired single cell RNA and TCR sequencing to assess the diversity and gene expression profile of these responses.

Conclusions: NEO-STIM is a novel platform to understand the immunogenic potential of high-quality neoantigen-targets in detail. Moreover, this platform can be utilized to generate T cell products for adoptive cell therapy for patients with a variety of solid and liquid tumors.

Citation Format: Divya Lenkala, Jessica Kohler, Brian McCarthy, Michael Nelson, Rachel DeBarge, Yvonne Ware, Yuting Huang, Janani Sridar, Yusuf Nasrullah, Dewi Hartjano, Asaf Poran, Sejuti Sengupta, Joost H. van den Berg, Matt Goldstein, Richard B. Gaynor, Marit M. van Buuren. Detailed analysis of immune responses induced against patient-specific neoantigens using the ex-vivo induction protocol, NEO-STIM™ [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 580.
Abstract 582: Tumor immunoediting in a lung cancer mouse model harboring EGFR mutations
Authors: Nishii Kazuya , Ohashi Kadoaki , Ninomiya Kiichiro , Makimoto Go , Watanabe Hiromi , Kano Hirohisa , Hara Naofumi , Udono Heiichiro , Kiura Katsuyuki
Published: August 2019

Background: Anti-Programed cell death 1 (PD-1) antibodies such as nivolumab or pembrolizumab show promising treatment effects in smoking-related lung cancer. However, anti-PD-1 antibodies have very little effect on epidermal growth factor receptor (EGFR) mutant non-small cell lung cancer (NSCLC). EGFR mutations are the most frequent oncogenic drivers in non-smoking related NSCLC. Therefore, we aimed to explore the mechanism underlying tumor immunoediting in a lung cancer mouse model harboring EGFR mutations.

Methods: We previously established two strains of transgenic lung cancer mouse (C57BL/6) harboring a mouse Egfr exon 19-deletion (mDEL) or a human EGFR L858R (hLR) mutation, driven by a surfactant protein (SP)-C promoter (Ohashi, Cancer Sci. 2008; Ohashi, Cancer Res. 2009). These mice developed EGFR-dependent multifocal lung adenocarcinomas from type II pneumocytes. We modified the two models by transplanting lung tumors subcutaneously into C57BL/6 (Ohashi, AACR. 2018). Evaluation of mice tumor lymphocyte profile was done through flow cytometry or immunohistochemistry (IHC). Effects of T cell depletion or anti-mouse PD-1 antibody (4H2) in vivo were also assessed. Comprehensive analysis of mRNA expression of immuno-checkpoint molecules in the tumors was performed using PanCancer Immune Profiling Panel (NanoString Technologies).

Results: CD4+ T cells and/or CD8+ T cells were depleted in the subcutaneously transplanted tumor model followed by treatment with gefitinib (50 mg/kg/day, 5 days/week) for 2 weeks. As a result, after gefitinib discontinuation, rapid regrowth was seen in the tumors from groups with depletion of CD8+ T cells; depletion of CD4+ T cells had little effect on tumor regrowth, and tumor inhibition continued for 2 weeks after cessation of gefitinib. IHC showed relatively increased numbers of CD4+ T cells in the mice tumors whereas a decrease in the number of CD8+ T cells or Foxp3 positive cells was noted. PD-L1 expression was not observed in the tumors. Consistently, 4H2 showed little effect on tumor growth in vivo. The cancer immune panel showed increased expression of immune mediators such as Ccl6, Clec4n, and Lcn2.

Conclusions: Our results suggest that CD8+ T cells can recognize mice tumors in vivo. New immuno-checkpoint molecules regulating CD8+ T cells from PanCancer Immune Profiling Panel were evaluated. Further, we plan to evaluate the efficacy of various immuno-checkpoint inhibitors in vivo.

Citation Format: Kazuya Nishii, Kadoaki Ohashi, Kiichiro Ninomiya, Go Makimoto, Hiromi Watanabe, Hirohisa Kano, Naofumi Hara, Heiichiro Udono, Katsuyuki Kiura. Tumor immunoediting in a lung cancer mouse model harboring EGFR mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 582.
Abstract 592: Associations of mammographic breast density with breast stem cell marker-defined breast cancer subtypes
Authors: Yaghjyan Lusine , Esnakula Ashwini , Scott Christopher , Wijayabahu Akemi , Jensen Matthew , Vachon Celine
Published: August 2019

Background: High mammographic breast density is a strong, well-established breast cancer risk factor. A recent study found increased expression of stem cell markers in the areas of high breast density, but, whether stem cells may explain high breast cancer risk due to high breast density is unknown. We investigated the association between breast density and the risk of breast cancer by the status of stem cell markers CD44, CD24, and ALDH1A1 in the tumor.

Methods: We included 223 postmenopausal women with primary invasive or in-situ breast cancer and 399 age-matched controls from Mayo Clinic Mammography Study. Percent breast density, absolute dense area and non-dense area were assessed using computer-assisted thresholding technique. Immunohistochemical analysis of the markers was performed on tumor tissue microarrays according to a standard protocol with commercial antibodies. Cases with positive staining (≥1% positively stained cells) on one or more cores were considered marker-positive. We used polychotomous logistic regression to quantify the associations of breast density measures with breast cancer risk by the status of each of the markers. Heterogeneity tests were conducted to compare the differences in associations by marker-defined tumor subtypes.

Results: Of the 223 cancers in the study, 182 (82%) were positive for CD44, 83 (37%) for CD24 and 52 (23%) for ALDH1A1. Associations of percent density were not significantly different across any of the marker-defined subtypes (density 51%+ vs. 11-25%: OR=2.83, 95% CI 1.49-5.37 for CD44+ vs. OR=1.87, 95% CI 0.47-7.51 for CD44-, p-heterogeneity=0.66; OR=2.80, 95% CI 1.27-6.18 for CD24+ vs. OR=2.44, 95% CI 1.14-5.22 for CD24-, p-heterogeneity=0.61; OR=3.04, 95% CI 1.14-8.10 for ALDH1A1+ vs. OR=2.57. 95% CI 1.30-5.08 for ALDH1A1-, p-heterogeneity=0.94). Positive associations of absolute dense area and inverse associations of non-dense area with breast cancer risk were similar across marker-defined subtypes. Similar associations of breast density measures with breast cancer were seen when cancer subtypes were defined using combined marker expression status (none, one positive marker, or two and more positive markers).

Conclusions: We found no evidence of differential associations of breast density measures with breast cancer risk by the status of stem cell markers. However, our power was limited and further studies in larger study populations are warranted to confirm these associations in a larger sample.

Citation Format: Lusine Yaghjyan, Ashwini Esnakula, Christopher Scott, Akemi Wijayabahu, Matthew Jensen, Celine Vachon. Associations of mammographic breast density with breast stem cell marker-defined breast cancer subtypes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 592.
Abstract 602: Biological monitoring to clarify potential causes of colorectal cancer in Korean diet
Authors: Yang Mihi , Lee Myungah , Kim Jeongseon Kim , Park Jong , Om Aeson , Damonte Jennifer
Published: August 2019

We performed a pilot study (N=30) to look for the Korean specific risk factors, which can explain an increased incidence rate of colorectal cancer (CRC) among Korean population. Based on the promising data from the pilot study, we performed a case-control study (N=218, age: 64.4 ±13.4 years; N of men=116, 53.2%) with biological monitoring to analyze various biomarkers including clinical/hematological indices, oxidative or inflammatory biomarkers, e.g., c-reactive protein (CRP), malondialdehyde (MDA) or homocysteine, the metabolites of polycyclic aromatic hydrocarbons (PAHs) and heterocyclic amines (HCAs), e.g., urinary 1-OHP, MeIQx and PhIP, and their DNA adducts, e.g., dG-C8-MeIQx and -PhIP, with UPLC/MS/MS. We also obtained participants’ epidemiological, clinical, and behavioral information and developed food frequency questionnaires (FFQ) to collect their food information for one year before cancer diagnoses. Total subjects consumed 53.4±74.0g/day of red meat, and 1.1±3.7g/day of processed meat. After adjustment for sex and age, there were no associations between CRC risk and red or processed meat intake, or urinary MeIQx or PhIP levels. However, there were inverse associations between CRC and milk or calcium intake (ps<0.05). In addition, urinary MeIQx levels were positively associated with intake of lipid or coffee (p=0.06, p<0.01, respectively). LDL-C and CRP levels in the cases were higher than ones from controls (ps<0.05). Among biomarkers, MDA levels were positively correlated to urinary levels of 1-OHP, PhIP and MeIQx (ps<0.05). These relations support the fact that PAHs or HCAs indirectly induce oxidative stress. Finally, fruits and milk intake showed protective effects, however, gender (man), CRP, and lipid intake were risk factors to CRC, based on multivariable regression analyses. Therefore, total lipid intake, oxidative stress, or inflammation may increase risks of CRC in Korean population rather than red or processed meat intake.

Citation Format: Mihi Yang, Myungah Lee, Jeongseon Kim Kim, Jong Park, Aeson Om, Jennifer Damonte. Biological monitoring to clarify potential causes of colorectal cancer in Korean diet [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 602.
Abstract 605: Association of blood lipid profile and lung cancer risk: Results from the Southern Community Cohort Study
Authors: Sun Yan , Wu Jie , Yoon Hyung-Suk , Zheng Wei , Blot William J. , Cai Qiuyin
Published: August 2019

Lung cancer is the second most common cancer and the leading cause of cancer death in the U.S. African Americans (AAs) have a higher incidence rate than any other racial group. Lipid metabolism and homeostasis play important roles in the growth of cancer cells and the regulation of membrane proteins. Some previous epidemiological studies have evaluated the association between blood lipid levels and lung cancer risk; however, the results were inconsistent, and few explored patterns among AAs. To investigate the association of blood lipid levels and lung cancer risk, particularly in a low socioeconomic population and AAs, we conducted a nested case-control study using resources from the Southern Community Cohort Study, a well-characterized prospective cohort study of approximately 86,000 adult men and women, two-thirds of whom are AAs. Blood lipid profiles, including total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides (TRIG), and lipoprotein-a (Lp-a), were measured in 372 lung cancer cases and 721 matched controls. AAs had lower median levels of TRIG, and higher levels of HDL and Lp-a than European Americans (EAs). Conditional logistic regression analysis revealed an inverse association between TC and lung cancer risk, with an adjusted OR for the highest vs. lowest tertiles of 0.67 (95% CI: 0.45-0.99). This association was observed in both AAs and EAs, but was only statistically significant in EAs. Further stratified analysis found that this association was only significant in cases that were diagnosed within 2 years of blood collection. No significant associations of other lipid markers with lung cancer risk were observed. The findings from our study suggest that the inverse associations between TC and lung cancer risk more likely resulted from cancerous occurrence and progression and associated lifestyle changes rather than being indicative of causal markers.

Citation Format: Yan Sun, Jie Wu, Hyung-Suk Yoon, Wei Zheng, William J. Blot, Qiuyin Cai. Association of blood lipid profile and lung cancer risk: Results from the Southern Community Cohort Study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 605.
Abstract 608: Examining H. pylori core genes for epidemiological typing
Authors: Yu Guoqin
Published: August 2019

H. pylori is the major causative risk factor of a serious of diseases including gastric cancer. It is highly diverse. Conventional method of typing H. pylori has been based on seven MLST loci, which however are not sufficient to distinguish closely-related strains. High diverse core genes that are present in all H. pylori strains are potential good candidates for investigating genomic variation within closely-related strains, therefore improving our understanding of H. pylori -related disease epidemiology. In this study, we investigated the variation of the H. pylori genome, especially its core genes. We downloaded 80 H. pylori whole genome from National Center for Biotechnology Information (NCBI), and identified core genes (99% <= strains <= 100%: 840 genes), Soft core genes (95% <= strains < 99%: 185), Shell genes (15% <= strains < 95%: 933), Cloud genes (0% <= strains < 15%: 2313). Compared with other gene groups, we found that core genes have higher proportion of polymorphic sites, similar level of nucleotide diversity per site, higher recombination events per site, and lower number of gaps per site. In addition, core genes evolve slower than other gene groups with lower Tajima’s D value and nonsynonymous to synonymous ratio. Among the core and soft-core genes, we found that majority have low genetic diversity while a few showed higher diversity. We evaluated these high diverse core genes further in terms of their evolution, function and potential to serve as candidates to distinguish closely related H. pylori strains.

Note: This abstract was not presented at the meeting.

Citation Format: Guoqin Yu. Examining H. pylori core genes for epidemiological typing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 608.
Abstract 610: A longitudinal investigation of hematologic perturbations among permethrin-exposed pesticide applicators in the Biomarkers of Exposure and Effect in Agriculture Study
Authors: Shearer Joe , Freeman Laura Beane , Liu Danping , Andreotti Gabriella , Hamilton Jennifer , Happel Julie , Lynch Charles , Alavanja Michael , Hofmann Jonathan
Published: August 2019

Background: In the Agricultural Health Study, a large prospective cohort of pesticide applicators, high lifetime use of the insecticide permethrin was associated with an increased risk of multiple myeloma. However, the biological mechanisms through which this association occurs remain to be fully understood.

Methods: Hematologic perturbations were assessed among 33 permethrin-exposed male pesticide applicators enrolled in the Biomarkers of Exposure and Effect in Agriculture study, a molecular substudy within the Agricultural Health Study. Serial blood samples were collected from each participant coinciding with permethrin use including visits during the offseason, an exposure visit the day after use, and for 27 participants a follow-up visit approximately three weeks after use. Complete blood count with white blood cell differential and lymphocyte subsets were collected at each visit. Linear mixed models were used to assess the relationship between permethrin exposure and hematologic parameters, controlling for potential confounders including season of blood draw, age, body mass index, recent cold/infection, and use of other pesticides.

Results: A statistically significant increase in the adjusted mean immature granulocyte count (37%, 95% CI=6%, 76%) was observed the day after permethrin use compared to an offseason visit. Statistically significant (P<0.05) alterations in several red blood cell (RBC) parameters were also observed on the day after permethrin use, including decreased RBC count and hemoglobin and increased mean corpuscular volume and RBC distribution width-SD. Decreases in RBC count and hemoglobin, and increased levels of RBC distribution width-SD persisted approximately three weeks after permethrin use.

Conclusions: Altered hematologic parameters could be indicative of disrupted hematopoiesis, providing insights into the potential biologic plausibility of the observed association between permethrin use and multiple myeloma risk among pesticide applicators.

Impact/Significance: Given the widespread use of permethrin in both residential and commercial settings, our findings may have public health implications beyond occupationally exposed pesticide applicators.

Citation Format: Joe Shearer, Laura Beane Freeman, Danping Liu, Gabriella Andreotti, Jennifer Hamilton, Julie Happel, Charles Lynch, Michael Alavanja, Jonathan Hofmann. A longitudinal investigation of hematologic perturbations among permethrin-exposed pesticide applicators in the Biomarkers of Exposure and Effect in Agriculture Study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 610.
Abstract 612: Overexpression of PCDHB9 is correlated with tumor progression in esophageal cancer
Authors: Oue Naohide , Yamamoto Yuji , Asai Ryuichi , Sakamoto Naoya , Sentani Kazuhiro , Yasui Wataru
Published: August 2019

Genes encoding transmembrane proteins expressed specifically in cancer cell may be ideal biomarkers for diagnosis. Previously, we showed that overexpression of PCDHB9, which was identified as a transmembrane protein, promotes peritoneal metastasis and correlates with poor prognosis in patients with gastric cancer. However, expression of PCDHB9 in esophageal cancer remains unclear. In the present study, we investigated the expression and function of PCDHB9 in esophageal squamous cell carcinoma (ESCC). Immunohistochemical analysis demonstrated that 51 (40%) of 128 GC cases were positive for PCDHB9. PCDHB9 expression was correlated with T classification, N classification and tumor stage. There was no significant correlation between PCDHB9 expression and patient survival. Cell adhesion to fibronectin and invasion activity were significantly reduced by PCDHB9 knockdown. These results suggest that PCDHB9 may be a novel biomarker and therapeutic target for ESCC.

Citation Format: Naohide Oue, Yuji Yamamoto, Ryuichi Asai, Naoya Sakamoto, Kazuhiro Sentani, Wataru Yasui. Overexpression of PCDHB9 is correlated with tumor progression in esophageal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 612.
Abstract 616: Literature review and meta-analyses of the prevalence of microsatellite instability high (MSI-H) and deficient mismatch repair (dMMR) for colorectal (CRC), gastric (GC), endometrial (EC) and ovarian cancers (OC) in Chinese population
Authors: Liu Xiaoqiao , Fan Jiaqi , Liaw Kai-Li , Xu Mo , Zhou Yu , Amonkar Mayur , Zeng Gefei
Published: August 2019

Purpose Given the high prevalence of MSI-H/dMMR in gastrointestinal and gynecologic cancers we systematically collected published studies and investigated its’ prevalence for Chinese CRC, GC, EC, OC subjects to better understand the size of potential target population available for pembrolizumab treatment in China.

Experimental Design Both Chinese and English literature were searched in WanFang and PubMed database respectively. Studies were included if they assessed prevalence of MSI-H by PCR based method investigating 5 markers from NCI or Promega panel, and/or dMMR by immunohistochemistry (IHC) for all 4 MMR proteins. Studies focusing only on hereditary or sporadic cancer were filtered out. Among eligible studies, meta-analyses (DerSimonian Laird random effect model) were performed on the proportion of MSI-H/dMMR for each tumor type by R metafor package. Prevalence for advanced stage (III/IV) were also extracted if the data were available.

Results For CRC, the prevalence of MSI-H estimated from 11 studies (13.7% [10.2-17.6%]) was similar as that of dMMR from 22 studies (14.6% [12.3-17.1%]). Pooled prevalence of MSI-H/dMMR from 32 studies (12,341 subjects) was estimated at 14.1% (12.2-16.1%), and 9.7% (7.2-12.5%) from 17 studies (3,933 subjects) for advanced stage CRC. The prevalence estimation for GC, EC and OC were 18.5%, 26% and 12.9% respectively. Further, the prevalence in advanced stage cancer, regardless of the cancer type, tended to be numerically lower than the all-stage combined prevalence.

Conclusions This was the first study comprehensively integrating published studies to estimate prevalence of MSI-H/dMMR across 4 main tumor types for Chinese subjects. The meta-analysis indicated the prevalence of MSI-H/dMMR for all or advanced stage Chinese cancer patients might be similar as that for western population.

Citation Format: Xiaoqiao Liu, Jiaqi Fan, Kai-Li Liaw, Mo Xu, Yu Zhou, Mayur Amonkar, Gefei Zeng. Literature review and meta-analyses of the prevalence of microsatellite instability high (MSI-H) and deficient mismatch repair (dMMR) for colorectal (CRC), gastric (GC), endometrial (EC) and ovarian cancers (OC) in Chinese population [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 616.