bluuebunny/crossref_metadata_2025 huggingface dataset:
Optimization of Aeroengine Shop Visit Decisions Based on Remaining Useful Life and Stochastic Repair Time
Authors: Cai Jing , Li Xin , Chen Xi
Published: December 2016

Considering the wide application of condition-based maintenance in aeroengine maintenance practice, it becomes possible for aeroengines to carry out their preventive maintenance in just-in-time (JIT) manner by reasonably planning their shop visits (SVs). In this study, an approach is proposed to make aeroengine SV decisions following the concept of JIT. Firstly, a state space model (SSM) for aeroengine based on exhaust gas temperature margin is developed to predict the remaining useful life (RUL) of aeroengine. Secondly, the effect of SV decisions on risk and service level (SL) is analyzed, and an optimization of the aeroengine SV decisions based on RUL and stochastic repair time is performed to carry out JIT manner with the requirement of safety and SL. Finally, a case study considering two CFM-56 aeroengines is presented to demonstrate the proposed approach. The results show that predictive accuracy of RUL with SSM is higher than with linear regression, and the process of SV decisions is simple and feasible for airlines to improve the inventory management level of their aeroengines.
Analysis of the Damage Mechanism Related to CO2Laser Cochleostomy on Guinea Pig Cochlea
Authors: Liu Xiang , Qian Xiao-qing , Ma Rui , Chi Fang-Lu , Ren Dong-Dong
Published: December 2016

Different types of lasers have been used in inner ear surgery. Therefore, it is of the utmost importance to avoid damage to the inner ear (e.g., hyperthermia and acoustic effects) caused by the use of such lasers. The aim of this study was to use a high powered fibre-enabled CO2laser (10 W, 606 J/cm2) to perform cochleostomies on guinea pig cochlea and to investigate the possible laser-induced damage mechanisms. The temperature changes in the round window membrane, auditory evoked brainstem response, and morphological of the hair cells were measured and recorded before and after laser application. All of the outcomes differed in comparison with the control group. A rise in temperature and subsequent increased hearing loss were observed in animals that underwent surgery with a 10 W CO2laser. These findings correlated with increased injury to the cochlear ultrastructure and a higher positive expression of E-cadherin andβ-catenin in the damaged organ of Corti. We assume that enhanced cell-cell adhesion and the activatedβ-catenin-related canonical Wnt-signalling pathway may play a role in the protection of the cochlea to prevent further damage.
Abstract A04: Identification of novel pancreatic cancer-specific biomarkers with organoid models
Authors: Engle Dannielle , Tiriac Herve , Keith Rivera , Ludwig Michael , Hwang Chang-il , Yu Kenneth , Pappin Darryl , Tuveson David
Published: December 2016

Pancreatic ductal adenocarcinoma (PDA) is almost uniformly lethal and surgical intervention is the only cure. Unfortunately, the majority of patients are ineligible for resection because of the advanced stage of disease by the time of diagnosis. This is due in part to that lack of diagnostic tools, especially for families with elevated risk. Although considered an intractable and aggressive malignancy, progression from the initiating events to overt cancer requires approximately 12 years, and advancement to metastatic disease takes another 5 – 7 years. This latency indicates that there is a window for early detection and intervention. The PDA biomarker, CA19-9, is measured in the blood to follow tumor burden longitudinally, but is neither sensitive nor specific enough to be used for diagnosis. In particular, the use of CA19-9 in PDA diagnosis is problematic given the elevation of CA19-9 in benign conditions, such as pancreatitis, that have a higher incidence than PDA. To improve the diagnostic performance of CA19-9, we identified tissue- and cancer-specific protein carriers of the CA19-9 carbohydrate modification using murine and human organoid model systems of pancreatic cancer disease progression. To induce CA19-9 expression in murine organoids, two human gylcosyl transferases that were deficient in mice were ectopically expressed. CA19-9 protein carriers specific to malignancy were identified relative to normal, proliferating ductal cells. Some CA19-9 carriers that were shared between normal and malignant pancreatic ductal organoids were also detected in the sera of patients with benign pancreatic disease, highlighting their inability to distinguish malignancy from pancreatitis. Importantly, several of the tumor-specific novel CA19-9 carriers identified in organoids also discriminated between benign and malignant disease in human patient sera, and are nominated for clinical validation. Overall, organoid model systems allow identification of biomarkers that discriminate between normal proliferation and malignant conditions of multiple gastrointestinal tissues, providing a robust platform for biomarker discovery.

Citation Format: Dannielle Engle, Herve Tiriac, Rivera Keith, Michael Ludwig, Chang-il Hwang, Kenneth Yu, Darryl Pappin, David Tuveson.{Authors}. Identification of novel pancreatic cancer-specific biomarkers with organoid models. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A04.
Abstract A07: Identification of endocrine/metabolic biomarkers associated to early PDC in a transgenic mouse model
Authors: Pasquale Valentina , Dugnani Erica , Liberati Daniela , Marra Paolo , Canu Tamara , Esposito Antonio , Piemonti1 Lorenzo
Published: December 2016

Background: Pancreatic ductal adenocarcinoma (PDC) is a deadly disease, even surgery contributes little to survival if PDC is not diagnosed at an initial stage. Glucose homeostasis alteration associated to PDC was described to identify an high risk population for screening. Moreover the weight loss often manifests several months before cachexia. The crosstalk between metabolism and cancer could be a key to early diagnosis. Our aim is to find endocrine/metabolic biomarkers of early PDC development in a gold standard murine model.

Materials and Methods: Candidate markers were investigated using LSL-KrasG12D/+;LSL-Trp53R172H/+;Pdx-1-Cre (KPC) mouse, a genetically-engineered model that develops PDC with 100% penetrance. KPC has been fed with standard diet (SD) or high fat diet (HFD) introducing a further risk factor for diabetes and metabolic alterations. To overcome the problem of asynchronous development of PDC in KPC, 7T-MRI was used to classify mice in actual pathological stage: normal (S1); cystic degeneration (S2); small tumor (S3) and advanced pancreatic cancer (S4). We performed metabolic test (OGTT) and serum endocrine/metabolic profile (xMAP technology and clinical biochemistry) in n=27 KPC (and control mice PDX-1 Cre) fed with HFD vs n=26 KPC (and control mice PDX-1 Cre) fed with SD. To prioritize markers we computed the ROC curve analysis.

Results: i) Glucose metabolism: KPC were normoglycemic regardless of the diet; the AUC in OGTT increases in KPC fed with HFD (HFD vs SD: S1 p<0,001, S2 p<0,001, S3 p=0,002, Cre p<0,001) ii) Serum levels of 29 endocrine/metabolic biomarkers were assessed during disease progression. The more relevant results are: increase of PYY (p<0,003) and decrease of leptin (p<0,02), GIP (p<0.005) albumin (p<0.001) both in HFD and SD fed KPC (Cre vs S2); increase of amylase in stage 1 HFD-fed KPC (p=0,037). Moreover PYY was able to discriminate control from stage 2 KPC with an AUC=0,929 in the ROC curve analysis (p<0,001).

Conclusions: Unlike human, in the mouse model the development of PDC is not associated with an hyperglycemic status, regardless of the diet. However KPC fed with HFD experience a mild impairment of glucose metabolism (as well as in control mice) and a mild pancreatic damage. Our results identify Leptin, PYY, GIP and albumin as markers of early stage PDC in a preclinical model with a diagnostic potential to be validated in human patients.

Citation Format: Valentina Pasquale, Erica Dugnani, Daniela Liberati, Paolo Marra, Tamara Canu, Antonio Esposito, Lorenzo Piemonti1.{Authors}. Identification of endocrine/metabolic biomarkers associated to early PDC in a transgenic mouse model. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A07.
Abstract A34: Oncogenic KRAS regulates pancreatic cancer cell signaling via stromal reciprocation
Authors: Tape Christopher , Ling Stephanie , Dimitriadi Maria , McMahon Kelly , Worboys Jonathan , Leong Hui S. , Norrie Ida , Miller Crispin , Poulogiannis George , Lauffenburger Douglas , Jorgensen Claus
Published: December 2016

Pancreatic Ductal Adenocarcinoma is characterized by a reactive stroma, which modifies tumor progression and response to therapy. Oncogenic mutations regulate signaling both within tumor cells and adjacent stromal cells. However, defining whether oncogenes can regulate tumor cell signaling and phenotypic behavior via stromal cells is of importance to understand mechanisms of disease progression and response to therapy. Here we show that oncogenic KRAS (KRASG12D) regulates pancreatic cancer cell signaling via stromal stellate cells. By combining cell-specific proteome labeling with multivariate phosphoproteomics, we analyzed heterocellular KRASG12D signaling in Pancreatic Ductal Adenocarcinoma (PDA) cells. Tumor cell KRASG12D engages heterotypic stellate cells, which subsequently instigate reciprocal signaling in the tumor cells. Reciprocal signaling employs additional kinases and doubles the number of regulated signaling nodes from cell-autonomous KRASG12D. Consequently, reciprocal KRASG12D produces a tumor cell phosphoproteome and total proteome that is distinct from cell-autonomous KRASG12D alone. Reciprocal signaling regulates tumor cell proliferation, apoptosis, and increases mitochondrial capacity via an IGF1R/AXL-AKT axis. These results demonstrate that oncogene signaling should be viewed as a heterocellular process and that our existing cell-autonomous perspective underrepresents the extent of oncogene signaling in cancer.

Citation Format: Christopher Tape, Stephanie Ling, Maria Dimitriadi, Kelly McMahon, Jonathan Worboys, Hui S. Leong, Ida Norrie, Crispin Miller, George Poulogiannis, Douglas Lauffenburger, Claus Jorgensen.{Authors}. Oncogenic KRAS regulates pancreatic cancer cell signaling via stromal reciprocation. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A34.
Abstract A37: Targeting RAGE in pancreatic cancer using monoclonal antibodies
Authors: Swami Priyanka , Indurthi Venkata , Vetter Stefan W. , Leclerc Estelle
Published: December 2016

Recent studies suggest that the Receptor for Advanced Glycation End Products (RAGE) and its ligands (S100P or HMGB1) are important contributors to the progression of pancreatic cancer. Being a cell surface receptor, RAGE possesses a large extracellular domain that binds its activating ligands. We reasoned that monoclonal antibodies that block the interaction between RAGE and its ligands could impact the progression of pancreatic cancer. We have generated a panel of antibodies recognizing different domains of the receptor and are currently testing their inhibitory effects, in pancreatic cancer cells and in small animal models.

Citation Format: Priyanka Swami, Venkata Indurthi, Stefan W. Vetter, Estelle Leclerc.{Authors}. Targeting RAGE in pancreatic cancer using monoclonal antibodies. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A37.
Abstract A51: Notch4 acts as an oncogenic signal in pancreatic tumorigenesis
Authors: Qiu Wanglong , Suh David D. , Tsay Natasha M. , Chadi Simona , Chambers Adam R. , Sims Peter A. , Shawber Carrie J. , Kitajewski Jan , Remotti Helen E. , Su Gloria H.
Published: December 2016

The Notch signaling network is an evolutionarily conserved intercellular signaling pathway which regulates interactions between adjacent cells. The Notch family receptor and its signaling pathways are involved in the development of numerous tissues in multicellular organisms. Recent studies from our Acvr1bflox/flox;LSL-KrasG12D;Pdx1-Cre mice suggest that Notch4 plays an oncogenic role in dictating cell fate and contributes to pancreatic tumorigenesis, particularly in the development of pancreatic intraductal papillary mucious neoplasms (IPMNs). The perinuclear localization of the activated Notch4 (but not other Notch members) intracellular domain (ICD) in the apical cytoplasm of neoplastic cells appeared to be associated with the expansion of the IPMN lesions. The unique role of Notch4 signaling pathway in IPMN development was also confirmed by unbiased RNA-Seq analyses. The deletion of Notch4 in the p16flox/flox;LSL-KrasG12D;Pdx1-Cre mice prolonged survival, further supporting its oncogenic role in pancreatic tumorigenesis.

Citation Format: Wanglong Qiu, David D. Suh, Natasha M. Tsay, Simona Chadi, Adam R. Chambers, Peter A. Sims, Carrie J. Shawber, Jan Kitajewski, Helen E. Remotti, Gloria H. Su.{Authors}. Notch4 acts as an oncogenic signal in pancreatic tumorigenesis. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A51.
Abstract A72: Phase 1 study of nivolumab (nivo) + nab-paclitaxel (nab-P) ± gemcitabine (G) in pancreatic cancer (PC): safety evaluation of patients treated with nivo + nab-P in arm A
Authors: Hochster Howard S. , Wainberg Zev A. , Gutierrez Martin , Waterhouse David , Chiorean E Gabriela , George Ben , Ko Amy , Manax Victoria , Stergiopoulos Sotirios , O'Dwyer Peter
Published: December 2016

Background: Cytotoxic chemotherapy activates the immune system through multiple mechanisms, including stress signaling, antigen release by dying tumor cells, and modulation of immune cell populations. Nivo, an anti–PD-1 antibody, has demonstrated activity in other solid tumors but has not yet demonstrated activity in PC. The combination of nab-P + G is approved by the US Food and Drug Administration as first-line therapy for metastatic PC (MPC). nab-P does not require corticosteroid pretreatment, which makes it a suitable combination partner with nivo to test for safety and efficacy in PC. Here we report interim results from the first of 2 PC cohorts of a Phase 1 study of nivo + the standard dose and schedule of nab-P.

Methods: The 2 PC cohorts (arms A and B) were to be initiated sequentially. In arm A, patients with locally advanced PC (LAPC) or MPC who had received 1 prior chemotherapy regimen received nab-P 125 mg/m2 on days 1, 8, and 15 of a 28-day cycle (qw 3/4) in combination with nivo 3 mg/kg on days 1 and 15 of the 28-day cycle starting with cycle 1. The primary objective of part 1 was to identify dose-limiting toxicities (DLTs). Patients treated with ≥ 2 cycles of nivo + nab-P or who discontinued due to a DLT prior to completing 2 cycles of nivo + nab-P were considered DLT evaluable. If arm A is deemed safe per DLT evaluation, arm B will enroll chemotherapy-naive patients, who will receive G 1000 mg/m2 qw 3/4 in addition to nivo + nab-P.

Results: Between February 24 and December 3, 2015, 11 patients were enrolled in arm A. Most patients were < 75 years of age (82%) and male (55%). No DLTs were observed, and no grade 3/4 AEs occurred in > 1 patient. Among 8 response-evaluable patients, 1 had a partial response and 4 achieved stable disease. Eight patients discontinued treatment due to disease progression.

Conclusions: In arm A of the PC cohort, the addition of nivo to nab-P did not appear to increase safety/tolerability concerns. Based on safety observed in arm A, arm B will enroll patients with MPC for first-line treatment with nivo + nab-P + G. ClinicalTrials.gov: NCT02309177.

Citation Format: Howard S. Hochster, Zev A. Wainberg, Martin Gutierrez, David Waterhouse, E Gabriela Chiorean, Ben George, Amy Ko, Victoria Manax, Sotirios Stergiopoulos, Peter O’Dwyer.{Authors}. Phase 1 study of nivolumab (nivo) + nab-paclitaxel (nab-P) ± gemcitabine (G) in pancreatic cancer (PC): safety evaluation of patients treated with nivo + nab-P in arm A. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A72.
Abstract A77: Screening of conditionally reprogrammed patient-derived carcinoma cells identifies ERCC3-MYC interactions as a target in pancreatic cancer
Authors: Astsaturov Igor , Jablonski Sandra A. , Zhou Yan , Serebriiskii Ilya , Keren Paz , Weiner Louis M. , Golemis Erica , Khazak Vladimir
Published: December 2016

Methods: Pancreatic ductal adenocarcinoma (PDAC) is a devastating malignancy that affects 44,000 individuals yearly in the US. The list of agents active against PDAC is limited. We explored in an unbiased fashion the clinically available drugs to identify novel cytotoxics using new physiological models for PDAC including cells rapidly expanded in vitro from surgical or xenografted tumors (PDX).

Results: Not unexpectedly, only a small minority of agents showed activity against pancreatic carcinoma cells in vitro. Among these, transcriptional repressors and drugs interfering with protein folding and biosynthesis ranked as most cytotoxic. Triptolide, a covalent inhibitor of the ERCC3, a bifunctional regulator of transcription and DNA repair, was most consistently effective in vitro and highly effective in vivo, causing prolonged complete regression in multiple PDX models. Importantly, triptolide showed superior activity in MYC-amplified PDX models, suggesting a critical role for ERCC3 in this subset. Triptolide elicited rapid and profound depletion of MYC oncoprotein, a transcriptional co-factor for ERCC3. Expression of ERCC3 was MYC-dependent, while resistance to triptolide was associated with elevated ERCC3 and MYC expression. Furthermore, high ERCC3 mRNA level was associated with decreased survival of PDAC patients.

Conclusions: These findings provide preclinical evidences for transcriptional vulnerability in PDAC via ERCC3 targeting and a new mechanistic approach for disruption of MYC-dependent pancreatic cancers.

Citation Format: Igor Astsaturov, Sandra A. Jablonski, Yan Zhou, Ilya Serebriiskii, Paz Keren, Louis M. Weiner, Erica Golemis, Vladimir Khazak.{Authors}. Screening of conditionally reprogrammed patient-derived carcinoma cells identifies ERCC3-MYC interactions as a target in pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A77.
Abstract B19: Kras mutation imparts neoplastic potential on duct cells but not acinar cells in a mouse model of obstructive chronic pancreatitis
Authors: Pan Fong C. , Kim Jessica N. , Shi Chanjuan , Washington Mary K. , Sander Maike , Gannon Maureen , Beauchamp Robert D. , Wright Christopher V. , Means Anna L.
Published: December 2016

Understanding the progression of human pancreatic cancer is difficult due to the late stage of diagnosis of this deadly disease. We must therefore rely upon robust model systems to understand how pancreatic cancer arises, how its acute and chronic phases of progression are regulated, and how best to identify and treat it, hopefully still in the early stage. Models of pancreatic ductal adenocarcinoma (PDAC) that accurately reflect human disease are still being developed. To mimic human disease, PDAC must arise from mutations occurring in adult animals and with etiologies relevant to humans. We developed a new mouse model that incorporates obstructive chronic pancreatitis with tissue-specific, adult-onset expression of mutant Kras. We found that KrasG12D expression in duct cells but not in acinar cells led to progression of metaplastic ducts to eventual dysplasia and cancer. In early-stage disease, Kras mutation in acinar cells led to increased acinar-to-ductal metaplasia but did not downregulate p53, thus leading to reduced cell survival. Ducts naturally express much lower p53 levels, however, and there was increased survival of KrasG12D-mediated, duct-derived metaplastic cells. Furthermore, acinar cells upregulated Pdx1 during acinar-to-ductal metaplasia while duct cells did not do so during their metaplastic transition. Without chronic pancreatitis, Kras mutation in acinar cells causes abundant Pancreatic Intraepithelial Neoplasm (PanIN)-like lesions. However, overexpression of Pdx1 in acinar cells in this context permitted acinar-to-ductal metaplasia but prevented PanIN-like lesion formation, suggesting that Pdx1 can repress neoplastic progression. In summary, in the setting of obstructive chronic pancreatitis, ducts are the principal source of cancer development via reduced Pdx1 and p53 levels and increased cell survival.

Citation Format: Fong C. Pan, Jessica N. Kim, Chanjuan Shi, Mary K. Washington, Maike Sander, Maureen Gannon, Robert D. Beauchamp, Christopher V. Wright, Anna L. Means.{Authors}. Kras mutation imparts neoplastic potential on duct cells but not acinar cells in a mouse model of obstructive chronic pancreatitis. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B19.
Abstract B20: The necrosome promotes pancreatic Oncogenesis via CXCL1 and mincle-induced immune Suppression
Authors: Seifert Lena , Werba Gregor , Tiwari Shaun , Giao Ly Nancy Ngoc , Alqunaibit Dalia , Alothman Sara , Daley Donnele , Hundeyin Mautin , Mani Vishnu R. , Barilla Rocky , Miller George
Published: December 2016

Neoplastic pancreatic epithelial cells are widely believed to die via Caspase 8-dependantapoptotic cell death and chemotherapy is thought to further promote tumor apoptosis. Conversely, disruption of apoptosis is a basic modality cancer cells exploit for survival. However, the role of necroptosis, or programmed necrosis, in pancreatic ductal adenocarcinoma (PDA) is uncertain. There are a multitude of potential inducers of necroptosis in PDA including ligation of TNFR1, CD95, TRAIL receptors, Toll-like receptors, ROS, and Chemotherapeutics. We doscovered that the principal components of the necrosome, RIP1 and RIP3, are highly expressed in PDA and are further upregulated by chemotherapy. Blockade of the necrosome in vitro promoted cancer cell proliferation and induced an aggressive oncogenic phenotype. However, belying these findings, in vivo RIP3 deletion or RIP1 inhibition was protective against oncogenic progression and was associated with the development of a highly immunogenic myeloid and T cell infiltrate. The immune-suppressive tumor microenvironment (TME) associated with intact RIP1/RIP3 signaling was in-part contingent on necroptosis-induced CXCL1 expression whereas CXCL1 blockade was protective against PDA. Moreover, we found that cytoplasmic SAP130 was expressed in PDA in a RIP1/RIP3-dependent manner, and Mincle – its cognate receptor – was upregulated in tumor-infiltrating myeloid cells. Mincle ligation by SAP130 promoted oncogenesis whereas Mincle deletion was protective and phenocopied the immunogenic reprogramming of the TME characteristic of RIP3 deletion. Cellular depletion experiments suggested that whereas inhibitory macrophages promote tumorigenesis in PDA, they lose their immune-suppressive effects in the context of RIP3 or Mincle deletion. As such, T cells which are dispensable to PDA progression in hosts with intact RIP3 or Mincle signaling become reprogrammed into indispensable mediators of anti-tumor immunity in absence of RIP3 or Mincle. Our work describes parallel networks of necroptosis-induced CXCL1 and Mincle signaling which critically promote macrophage-induced adaptive immune suppression enabling PDA progression.

Citation Format: Lena Seifert, Gregor Werba, Shaun Tiwari, Ly Nancy Ngoc Giao, Dalia Alqunaibit, Sara Alothman, Donnele Daley, Mautin Hundeyin, Vishnu R. Mani, Rocky Barilla, George Miller.{Authors}. The necrosome promotes pancreatic Oncogenesis via CXCL1 and mincle-induced immune Suppression. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B20.
Abstract B25: Activation of Wnt/β-catenin signaling pathway enhances pancreatic cancer development and the malignant potential via up-regulation of Cyr61/CCN1
Authors: Sano Makoto , Driscoll David R. , DeJesus-Monge Wilfredo E. , Quattrochi Brian , Appleman Victoria A. , Ou Jianhong , Zhu Lihua Julie , Yoshida Nao , Yamazaki Shintaro , Takayama Tadatoshi , Sugitani Masahiko , Nemoto Norimichi , Klimstra David S. , Lewis Brian C.
Published: December 2016

Pancreatic ductal adenocarcinoma (PDAC), a poor prognostic cancer, commonly develops following activating mutations in the KRAS oncogene. Activation of the Wnt signaling pathway is also commonly observed in PDAC. To ascertain the impact of postnatal activation of the Wnt signaling pathway in PDAC development, we combined the elastase-tva-based RCAS-TVA pancreatic cancer model with the established LSL-KrasG12D, Ptf1a-cre model. Delivery of RCAS viruses encoding β-cateninS37A and Wnt1 stimulated the progression of premalignant PanIN and PDAC development. Indeed, mice injected with RCAS-β-cateninS37A and Wnt1 had reduced survival relative to RCAS-GFP controls (P < 0.05). Active β-catenin or its DNA-binding partner TCF4 enhanced PDAC cell proliferation, soft-agar colony formation, as well as migration and invasion activity. In contrast, these phenotypes were significantly blocked by the introduction of Icat, an inhibitor of the β-catenin/TCF4 interaction. Meanwhile, we identified Cyr61/CCN1 as a target molecule for Wnt/β-catenin signaling pathway in PDAC. Nuclear β-catenin and Cyr61/CCN1 expression were predominantly detected in moderately to poorly differentiated murine and human PDAC. Indeed, nuclear β-catenin and Cyr61/CCN1-positivity independently correlated with poor prognosis (P < 0.01). Knockdown of Cyr61/CCN1 in a β-catenin-activated PDAC cell line reduced soft agar, migration and invasion activity. Together, these data suggest that the Wnt/β-catenin signaling pathway enhances pancreatic cancer development and malignancy in part via up-regulation of Cyr61/CCN1.

Citation Format: Makoto Sano, David R. Driscoll, Wilfredo E. DeJesus-Monge, Brian Quattrochi, Victoria A. Appleman, Jianhong Ou, Lihua Julie Zhu, Nao Yoshida, Shintaro Yamazaki, Tadatoshi Takayama, Masahiko Sugitani, Norimichi Nemoto, David S. Klimstra, Brian C. Lewis.{Authors}. Activation of Wnt/β-catenin signaling pathway enhances pancreatic cancer development and the malignant potential via up-regulation of Cyr61/CCN1. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B25.
Abstract B28: Dynamic interplay between type I collagen signaling and invasion in pancreatic cancer
Authors: Shields Mario A. , Maguin Pascal , Dallas Sarah L. , Egeblad Mikala
Published: December 2016

The stroma of pancreatic cancer can restrict as well as promote tumor progression. Why the stroma exhibits these paradoxical functions remains unclear. One of the major components of the pancreatic cancer stroma is fibrillar type I collagen. High levels of collagen in primary tumor specimens have been associated with poor survival and resistance to therapy in pancreatic cancer patients.

To determine the effects of type I collagen on cancer cell behavior, we performed intravital confocal microscopic imaging in mice. We used mice that expressed GFPtopaz fused to the α2 chain of type I collagen under control of a 3.6kb fragment of the COL1A1 promoter [α2(I)-col-GFP mice]. Live imaging of pancreatic tumors, formed after orthotopic injection of cells derived from the classical KPC mouse models, showed that type I collagen architecture in the tumors was heterogeneous. Strikingly, we observed that invasive cancer cells migrated out of the tumors on linear type I collagen. Linearization of collagen fibers through cross-linking, catalyzed by lysyl oxidases (LOXs), is inversely correlated with metastasis-free survival in breast cancer. However, in our pancreatic cancer model, inhibiting LOX promoted collagen linearization, tumor growth and metastatic spread. Elevated collagen-mediated signaling, through activation of focal adhesion kinase (FAK) and discoidin domain receptor (DDR1), was also observed in the tumors treated with lysyl oxidase inhibitor. Lysyl oxidase inhibition also promoted cell contractility as shown by elevated phosphorylated mysosin light chain 2 (p-MLC2) in vivo and by in collagen contractility assays in vitro. TGF-β signaling is a potent stimulator of collagen secretion and of invasive behavior, and it is aberrantly regulated in pancreatic cancer. We observed that lysyl oxidase inhibitor increased phosphorylated Smad3, a downstream effector of TGF-β signaling. Together, our data suggest that the influence of collagen on tumor behavior depends on its architecture. In areas of linear collagen, we find high cancer cell motility out of the tumors and this correlates with increased FAK and TGF-β signaling activity. The architecture-specific function of collagen could potentially explain the paradoxical role for the stroma in pancreatic cancer progression.

Citation Format: Mario A. Shields, Pascal Maguin, Sarah L. Dallas, Mikala Egeblad.{Authors}. Dynamic interplay between type I collagen signaling and invasion in pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B28.
Abstract B38: Clinical translation of nuclear export inhibitor in metastatic pancreatic cancer
Authors: Azmi Asfar S. , Muqbil Irfana , Aboukameel Amro , Kauffman Michael , Shacham Sharon , Mohammad Ramzi M. , Philip Philip A.
Published: December 2016

Pancreatic ductal adenocarcinoma (PDAC) remains a deadly disease in urgent need of newer therapeutic modalities. PDAC tumors are very heterogeneous and carry alterations in many critical pathways rendering the design of therapy against a single pathway unrealistic. The disease requires a broad form of therapy that can target the activation of multiple tumor suppressor proteins (TSPs) simultaneously. We have shown that over-expression of the nuclear exporter protein CRM1 in PDAC leads to functional inactivation of TSPs (such as FOXO3a, p27, Par-4 and p53) through their mislocalization to the cytoplasmic compartment. We clearly demonstrated that targeted inhibition of CRM1 by CRISPR/Cas9 validated Specific Inhibitor of Nuclear Export (SINE) KPT-330/Selinexor can restore the function of multiple TSPs leading to PDAC cell death and tumor inhibition in orthotopic models. More significantly, our findings show that SINE synergize with gemcitabine (GEM) and nab-paclitaxel leading to enhanced PDAC growth inhibition, apoptosis, and spheroid disintegration of PDAC derived cancer stem cells (CSCs) that undergo epithelial-to-mesenchymal transition (EMT). The observed synergy was due in part to enhanced nuclear localization of TSPs and suppression of CSC markers alongside reversal of EMT markers. RNA from quadruplet combination treatment samples were subjected to Agilent HT12 microarrays and differentially expressed genes (p<0.01) were subjected to pathway analysis. Our computational analysis demonstrated reversal of immune suppressive networks and suppression of CSC sustaining networks. Most importantly we observed activation of fibroblast specific cell death pathways highlighting an as of yet unexplored role of CRM1 in sustaining fibroblast growth that is recognized to support high desmoplastic reaction (DR) in PDAC. Our multi-model pre-clinical work has led to the approval of a Phase Ib/II clinical study involving GEM-nab-paclitaxel-Selinexor for metastatic PDAC (NCT02178436).

Citation Format: Asfar S. Azmi, Irfana Muqbil, Amro Aboukameel, Michael Kauffman, Sharon Shacham, Ramzi M. Mohammad, Philip A. Philip.{Authors}. Clinical translation of nuclear export inhibitor in metastatic pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B38.
Abstract B59: Black raspberries inhibit pancreatic carcinogenesis by suppressing Raf/MEK/ERK/STAT3 signaling pathways and promoting apoptosis
Authors: Pan Pan , Skaer Chad W. , Wang Hsin-Tzu , Tsai Susan , Oshima Kiyoko , Huang Yi-Wen , Wang Li-Shu
Published: December 2016

Background: Despite improved understanding of the pathophysiology of pancreatic ductal adenocarcinoma (PDAC) in the past two decades, PDAC has an extremely low 5-year survival rate (7%). However, diets rich in fruits and vegetables have been reported to reduce the risk of cancer development. We previously demonstrated that black raspberries (BRBs) have chemopreventive effects in human colorectal cancer patients and mouse models of colorectal cancer. Our current study aimed to investigate the potential effects of BRBs against PDAC in mice.

Methods: Mice that carry a constitutively activated KrasLSL.G12D/+ oncogene and loss-of-function Trp53LSL.R172H/+ tumor-suppressor gene in the pancreas (KrasLSL.G12D/+-Trp53LSL.R172H/+-Pdx-1-Cre) spontaneously develop PDAC that recapitulates the human PDAC. Four-week-old KrasLSL.G12D/+-Trp53LSL.R172H/+-Pdx-1-Cre mice were given a control diet or a control diet supplemented with 5% BRBs. In addition, luciferase-transfected human Panc-1 (Panc-1-Luc) cells were orthotopically injected into the pancreas of nonobese, diabetic, severe combined immunodeficient (NOD.SCID) mice. Four weeks after the inoculation of Panc-1-Luc cells, the NOD.SCID mice were given a control diet or a control diet supplemented with 5% BRBs for 6 weeks. Gross tumor volume of orthotopic pancreatic tumors was measured.

Results: The 4-week-old KrasLSL.G12D/+-Trp53LSL.R172H/+-Pdx-1-Cre mice developed pancreatic intraepithelial neoplasia (PanIN) lesions, which are the precancerous lesions of PDAC, and they were fed control or 5% BRB diet. Kaplan-Meier survival analysis showed that 5% BRBs significantly prolonged the survival of those PDAC mice with a median survival of 189 days versus 154 days for the mice fed the control diet. Molecular studies suggested that 5% BRBs suppress the Raf/MEK/ERK/STAT3 pathway, which is the downstream of Kras activation, and inhibit cancer cell proliferation in pancreatic tumor tissues. In addition, 5% BRBs significantly decreased the gross tumor volume of orthotopic tumors produced by injecting Panc-1-Luc cells into the pancreas. Orthotopic pancreatic tumors in BRB-treated NOD.SCID mice had a higher rate of apoptosis than tumors from mice fed the control diet.

Conclusions: These results support the hypothesis that BRBs have the clinical potential to delay pancreatic cancer progression by suppressing cancer cell proliferation and/or promoting apoptosis.

Citation Format: Pan Pan, Chad W. Skaer, Hsin-Tzu Wang, Susan Tsai, Kiyoko Oshima, Yi-Wen Huang, Li-Shu Wang.{Authors}. Black raspberries inhibit pancreatic carcinogenesis by suppressing Raf/MEK/ERK/STAT3 signaling pathways and promoting apoptosis. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B59.
Abstract B64: Using pancreatic organoids to infer therapeutic resistance and sensitivity
Authors: Tiriac Herve* , Ponz-Sarvise Mariano* , Corbo Vincenzo* , Frese Kristopher K.* , Engle Dannielle D.* , Ohlund Daniel , Oni Tobiloba , Hwang Chang-il , Handly-Santana Abram , Alagesan Brinda , Filippini Dea , Wright Kevin , Yu Kenneth H. , Park Youngkyu , Tuveson David A.
Published: December 2016

Pancreatic ductal adenocarcinoma (PDA) is a lethal disease with a 5-year survival rate less than 6%. The late diagnosis of the disease and poor efficacy of systemic treatments are major factors for this poor prognosis, and highlight the need for novel PDA models better able to predict resistance/sensitivity to treatment. To fulfill this need, our laboratory developed a three-dimensional culture system that enables the rapid growth of both mouse and human pancreatic ductal organoids. Organoids can be established from healthy or neoplastic tissues using both large (surgical material) and small (fine needle biopsy) samples, thereby providing a novel platform for therapeutic testing for all stages of disease. In this study, we use organoids to evaluate the therapeutic efficacy of the simultaneous inhibition of MEK and AKT kinases, which are activated in PDA by KRAS mutations. Tumor-derived organoids were resistant to both single agents and combination treatment. Genetically engineered mouse models of pancreatic cancer (KPC) also only exhibited a brief response to dual inhibition of MEK and AKT. To assess the potential mechanism of resistance, we evaluated the activation of a number of receptor tyrosine kinases by interrogating both mouse and organoid treated tissues. Members of the ERBB family were activated in KPC animals and tumor-derived organoids treated with both MEK and AKT inhibitors. Accordingly, the addition of an irreversible pan-ERBB inhibitor to the combination of MEK and AKT inhibitors, or MEK inhibitor alone, prevented ERBB-mediated resistance and improved tumor sensitivity to therapeutic intervention. This new therapeutic strategy is efficacious both ex-vivo and in vivo using human tumor-derived organoids and their associated transplant model. Finally, we show that human PDA organoids can be used in high throughput therapeutic screening for the identification of patient-specific therapeutic sensitivities. In summary, pancreatic organoids represent a system that can be used to predict drug sensitivity as well as to identify mechanism of resistance.

*Equal contribution

Citation Format: Herve Tiriac*, Mariano Ponz-Sarvise*, Vincenzo Corbo*, Kristopher K. Frese*, Dannielle D. Engle*, Daniel Ohlund, Tobiloba Oni, Chang-il Hwang, Abram Handly-Santana, Brinda Alagesan, Dea Filippini, Kevin Wright, Kenneth H. Yu, Youngkyu Park, David A. Tuveson.{Authors}. Using pancreatic organoids to infer therapeutic resistance and sensitivity. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B64.
Abstract B69: Tuft cell signaling can induce pancreatic inflammation and promote pancreatic ductal adenocarcinoma
Authors: Hoffman Megan T. , Gao Shan , Crawford Howard
Published: December 2016

The tuft cell is a chemosensory cell type that can interpret signals from luminal surfaces throughout the body in order to activate downstream effectors. One clear effector pathway of the tuft cell promotes a type-two immune response in the intestine after parasitic infection, suggesting that this cell population has the ability to communicate with the immune system. Furthermore, tuft cells have also been shown to activate sensory nerve fibers to lead to the production of a neuroinflammatory signal in the nasal epithelium. Therefore, tuft cells can promote effector inflammatory responses through direct immune cell signaling and through sensory nerve fibers. Pancreatic ductal adenocarcinoma (PDA) is characterized by a robust stromal reaction, a large component of which is composed of an immune response. It has been previously studied that this inflammatory response can function to promote both the initiation and progression of PDA. One mechanism by which inflammatory cells can be recruited to the pancreas is through neuroinflammatory pathways. Work with collaborators has shown that pancreatic sensory neurons are required for the progression of PDA, as ablation of this cell type significantly prolongs survival. Interestingly, tuft cells in the metaplastic pancreas have been shown to appear during transdifferentiation and progression to PDA. These tuft cells also possess key signaling elements that have the potential to mediate the response to extracellular signals, such as α-gustducin and TRPM5. Metaplastic tuft cells also produce the proteins necessary to make or release prostaglandins, acetylcholine and ATP, all of which can stimulate either a direct immunomodulatory signal or activate sensory nerve fibers. This leads us to hypothesize that tuft cells are a key mediator of signals to the stromal immune compartment and to sensory neurons in order to promote a robust immune response. In order to answer this question we are using chemosensory knockout animals to ablate critical signaling components of the chemosensory cascade to study the progression of PDA.

Citation Format: Megan T. Hoffman, Shan Gao, Howard Crawford.{Authors}. Tuft cell signaling can induce pancreatic inflammation and promote pancreatic ductal adenocarcinoma. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B69.
Abstract B70: RSPO2 Enhances Canonical Wnt Signaling to Confer Stemness-Associated Traits to Susceptible Pancreatic Cancer Cells
Authors: Ilmer Matthias , Wang Jing , Recio Alejandro , Werner Jens , Bazhin Alexandr , Alt Eckhard , Vykoukal Jody
Published: December 2016

Cancer stem cells (CSC) in pancreatic ductal adenocarcinomas (PDAC) present formidable challenges in the treatment of patients by escaping therapeutic intervention and seeding tumors through processes that remain incompletely understood. Here, we describe subpopulations of pancreatic cancer cells with high susceptibility to the recently described group of Wnt enhancer molecules, in particular R-Spondin 2 (RSPO2), which seem to drive primary and secondary pancreatic neoplasms.

PDAC cells responsive to RSPO2 stimuli upregulate canonical Wnt signaling activity as evidenced by Super-TOP/FOP luciferase assays and GFP-activity in cells transduced with a lentiviral Super-TOP-GFP reporter construct (7TGC). RSPO2-responsive cells (RSPO2pos) also possess properties indicative of CSCs, including tumor-initiating capacity and, more importantly, drug resistance in vivo. Cell populations with negligible Wnt activity preferentially express markers of differentiation. Furthermore, spontaneous response to extrinsic Wnt signals induced signaling networks comprising ERK1/2 and epithelial–mesenchymal transition (EMT) as shown by Western blot and RPPA analysis and led to a redistribution of Wnt enhancer receptors. In vivo, animals with RSPO2pos-derived tumors were bigger than RSPO2neg-derived tumors, harbored significantly more circulating tumor cells (CTCs), and higher number of exosomes in murine serum.

In this context, the latter findings suggest that RSPO2pos-cells could prepare secondary sites via exosomal priming and subsequently seed CSC-enriched CTCs for metastatic outgrowth at distant organs. RSPO2 gradients in the CSC niche might induce EMT for the developing CTCs. Hence, RSPO2 seems to play a prominent upstream role in regulating these interplays in the tumor and metastatic microenvironment.

In conclusion, our studies reveal adaptive aspects of pancreatic cancer stemness arising from populations of CSCs that misappropriate functional and responsive elements of archetypical self-renewal pathways. Blocking RSPO2-induced signaling pathways in conjunction with established chemotherapy could provide means to disrupt dynamic CSC processes and present novel therapeutic strategies for patients with PDAC.

Citation Format: Matthias Ilmer, Jing Wang, Alejandro Recio, Jens Werner, Alexandr Bazhin, Eckhard Alt, Jody Vykoukal.{Authors}. RSPO2 Enhances Canonical Wnt Signaling to Confer Stemness-Associated Traits to Susceptible Pancreatic Cancer Cells. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B70.
Abstract B73: Pancreatic stellate cell heterogeneity in cancer
Authors: Öhlund Daniel , Handly-Santana Abram , Elyada Ela , Biffi Giulia , Tuveson David A.
Published: December 2016

Paracrine interactions between pancreatic stellate cells (PSCs) and tumor cells play key roles in pancreatic cancer tumorigenesis. However, attempts to therapeutically target the PSCs have implied that PSCs may also restrain tumor growth, raising the possibility of diverse and dynamic pathophysiological roles of these cells in cancer. In order to gain a deeper understanding of the functional heterogeneity found among PSCs, we have developed an organotypic co-culture system that allows the study of functional interactions between pancreatic tumor-derived organoids and PSCs in a three-dimensional basement membrane matrix (Matrigel). PSCs embedded in Matrigel acquire a quiescent phenotype and restore their vitamin A droplets similar to PSCs found in healthy pancreata in vivo. In co-culture the PSCs proliferate, form lattices around the organoids, and secrete extracellular matrix proteins and inflammatory cytokines. In turn, the organoids proliferate more rapidly in co-culture with PSCs compared to their mono-cultured counterparts, as well as survive passaging in reduced media conditions.

Furthermore, we show that PSCs can differentiate into two distinct subtypes with different phenotypic features depending on the spatial proximity to the co-cultured organoids. PSCs found in direct contact with tumor cells express higher levels of smooth muscle actin (SMA), while PSCs that are more distant from the tumor cells show low SMA expression and gain a secretory phenotype. The secreted factors from this subtype of PSCs activate signaling pathways important for survival and proliferation in tumor organoids. We demonstrate that these two subtypes are mutually exclusive, and can both be found in vivo with similar spatial distribution as seen in the co-culture system.

Taken together, we have identified two subtypes of PSCs present in pancreatic cancer with potentially different pathophysiological functions. Preferentially targeting one of these subtypes in future clinical trials may positively affect patient outcomes.

Citation Format: Daniel Öhlund, Abram Handly-Santana, Ela Elyada, Giulia Biffi, David A. Tuveson.{Authors}. Pancreatic stellate cell heterogeneity in cancer. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr B73.