bluuebunny/crossref_metadata_2025 huggingface dataset:
Cardiopulmonary Resuscitation Pattern Evaluation Based on Ensemble Empirical Mode Decomposition Filter via Nonlinear Approaches
Authors: Sadrawi Muammar , Sun Wei-Zen , Ma Matthew Huei-Ming , Dai Chun-Yi , Abbod Maysam F. , Shieh Jiann-Shing
Published: July 2016

Good quality cardiopulmonary resuscitation (CPR) is the mainstay of treatment for managing patients with out-of-hospital cardiac arrest (OHCA). Assessment of the quality of the CPR delivered is now possible through the electrocardiography (ECG) signal that can be collected by an automated external defibrillator (AED). This study evaluates a nonlinear approximation of the CPR given to the asystole patients. The raw ECG signal is filtered using ensemble empirical mode decomposition (EEMD), and the CPR-related intrinsic mode functions (IMF) are chosen to be evaluated. In addition, sample entropy (SE), complexity index (CI), and detrended fluctuation algorithm (DFA) are collated and statistical analysis is performed using ANOVA. The primary outcome measure assessed is the patient survival rate after two hours. CPR pattern of 951 asystole patients was analyzed for quality of CPR delivered. There was no significant difference observed in the CPR-related IMFs peak-to-peak interval analysis for patients who are younger or older than 60 years of age, similarly to the amplitude difference evaluation for SE and DFA. However, there is a difference noted for the CI (p<0.05). The results show that patients group younger than 60 years have higher survival rate with high complexity of the CPR-IMFs amplitude differences.
Melioidosis: A Rare Cause of Liver Abscess
Authors: Martin Peter Franz M. San , Teh Catherine S. C. , Casupang Ma. Amornetta J.
Published: July 2016

Case Presentation. This is a case of a 44-year-old male, farmer, known to be diabetic, presenting with two-week history of vague abdominal pain associated with high grade fever. Abdominal CT scan showed localized liver abscess at segment 8 measuring 7.5 × 6.8 × 6.1 cm. Patient subsequently underwent laparoscopic ultrasound guided pigtail insertion for drainage of abscess. Culture studies showed moderate growth ofBurkholderia pseudomalleiin which the patient completed seven days of IV Meropenem. On follow-up after 12 weeks of oral Sulfamethoxazole/Trimethoprim, taken twice a day, the patient remained asymptomatic with no residual findings based on the abdominal ultrasound.Discussion. Diagnosis of melioidosis, a known “great masquerader,” relies heavily on culture studies. Consensus with regard to the management of liver abscess caused byBurkholderia pseudomalleihas not yet been established due to the rarity of cases. Surgical intervention through either a percutaneous or open drainage has shown good outcomes compared to IV antibiotics alone. In Philippines, the possibility of underreporting is highly plausible. This write-up serves not only to report a rare presentation of melioidosis but also to add to the number of cases reported in the country, possibly indicative of disease emergence.
Evaluation of the Efficiency of Two Different Freezing Media and Two Different Protocols to Preserve Human Spermatozoa from Cryoinjury
Authors: Fabozzi Gemma , Starita Maria Flavia , Rega Emilia , Alteri Alessandra , Colicchia Antonio , Piscitelli Claudio , Giannini Pierluigi
Published: July 2016

It is universally recognized that cryopreservation impairs sperm quality. In order to improve postthawing sperm survival and motility, media of different composition and different protocols have been proposed. However, no clear evidence is available to understand which are the most efficient protocol and medium for sperm cryopreservation. The present study evaluates the efficiency of two different cryopreservation protocols and two common freezing media (FM) containing different cryoprotectants (CPs), TEST Yolk Buffer (TYB) and Sperm Freeze (SF), to preserve human sperm quality. Our data suggest that TYB is better than SF both in terms of postthaw viability and in terms of progressive motility, while the direct addition of FM to the sperm sample resulted in the most efficient protocol in terms of postthaw viability but not in terms of progressive motility.
Factors Associated with Gastrointestinal Parasitic Infections among Young Population in Northeast Brazil
Authors: Silva Juliana Vasconcelos Lyra da , Fontes Gilberto , Santos Célia Dias dos , Santos Rafael Vital dos , da Rocha Eliana Maria Mauricio
Published: July 2016

Background. Intestinal parasitic infections constitute a major public health problem that is frequently associated with poverty, inadequate sanitation, and the nutritional status of the population.Objective. The aim of the present study is to investigate the possible association of parasitic infections, sanitary conditions, hygiene practices, and the nutritional and socioeconomic status of a poor youth population.Methods. A cross-sectional study was conducted with 367 children and adolescents inhabiting a substandard settlement in the urban area of Maceió (Alagoas State, Brazil). Data collection included socioeconomic status, anthropometric measurements, fecal sample examinations, and laboratory blood analysis. The identification of factors associated with gastrointestinal parasitic infections was undertaken through bi- and multivariate analyses.Results. Stool sample analysis obtained from 300 individuals revealed that 204 (68%) were infected with at least one parasite species and of these 130 (63.7%) were polyparasitized. No significant associations were identified between low height for age (stunted), parasitic infections, and polyparasitism. There was also no association between family income and parasitosis. However, low socioeconomic status proved to be a potential risk factor for parasitic infections.Conclusion. Actions must be taken to improve sanitation, housing, and environmental conditions in order to eliminate the risk factors for parasitic infections, and thereby guarantee a better quality of life for this population.
Caspian Sea Tidal Modelling Using Coastal Tide Gauge Data
Authors: Pirooznia Mahmoud , Rouhollah Emadi Seyyed , Najafi Alamdari Mehdi
Published: July 2016

The purpose of this paper is to model tidal conditions in the Caspian Sea using data from coastal tide gauges of Anzali, Noshahr, and Neka Ports. Harmonic analysis method was used to identify and examine 40 tidal components. The results illustrate that the annual (Sa) and semiannual solar (Ssa) components on all of the ports listed have the highest range in comparison with the other components which are, respectively, 16 cm, 18 cm, and 15 cm for annual components and 2.8 cm, 5.4 cm, and 3.7 cm for semiannual components.
The Role of Dynamic Contrast-Enhanced MRI in a Child with Sport-Induced Avascular Necrosis of the Scaphoid: A Case Report and Literature Review
Authors: Koc Baris Beytullah , Schotanus Martijn , Jong Bob , Tilman Pieter
Published: July 2016

Avascular necrosis (AVN) of the scaphoid in children is very rare and there is currently no consensus when conservative or operative treatment is indicated. A 10-year-old boy, practicing karate, presented with acute pain in his left wrist after falling on the outstretched hand. Imaging showed a scaphoid waist fracture with signs of an ongoing AVN. The diagnosis of AVN was confirmed with signal loss of the scaphoid on MRI T1. A dynamic contrast-enhanced MRI was performed for further assessment of the proximal pole vascularity and treatment planning. As dynamic contrast-enhanced MRI showed fair perfusion of the proximal pole, an adequate healing potential with conservative treatment was estimated. We achieved union and good function with cast immobilization for fourteen weeks. This case study showed dynamic contrast-enhanced MRI to be a valuable tool in assessing whether conservative or operative treatment is indicated to achieve union and good functional outcome.
Tyrosine Kinase Receptor Landscape in Lung Cancer: Therapeutical Implications
Authors: Quintanal-Villalonga A. , Paz-Ares Luis , Ferrer Irene , Molina-Pinelo S.
Published: July 2016

Lung cancer is a heterogeneous disease responsible for the most cases of cancer-related deaths. The majority of patients are clinically diagnosed at advanced stages, with a poor survival rate. For this reason, the identification of oncodrivers and novel biomarkers is decisive for the future clinical management of this pathology. The rise of high throughput technologies popularly referred to as “omics” has accelerated the discovery of new biomarkers and drivers for this pathology. Within them, tyrosine kinase receptors (TKRs) have proven to be of importance as diagnostic, prognostic, and predictive tools and, due to their molecular nature, as therapeutic targets. Along this review, the role of TKRs in the different lung cancer histologies, research on improvement of anti-TKR therapy, and the current approaches to manage anti-TKR resistance will be discussed.
A Phase I Trial to Evaluate Antibody-Dependent Cellular Cytotoxicity of Cetuximab and Lenalidomide in Advanced Colorectal and Head and Neck Cancer
Authors: Bertino Erin M. , McMichael Elizabeth L. , Mo Xiaokui , Trikha Prashant , Davis Melanie , Paul Bonnie , Grever Michael , Carson William E. , Otterson Gregory A.
Published: July 2016

mAbs can induce antibody-dependent cellular cytotoxicity (ADCC) via the innate immune system's ability to recognize mAb-coated cancer cells and activate immune effector cells. Lenalidomide is an immunomodulatory agent with the capacity to stimulate immune cell cytokine production and ADCC activity. This phase I trial evaluated the combination of cetuximab with lenalidomide for the treatment of advanced colorectal and head and neck squamous cell cancers (HNSCC). This trial included patients with advanced colorectal cancer or HNSCC. Treatment consisted of cetuximab 500 mg/m2 i.v. every two weeks with lenalidomide given orally days 1–21 on a 28-day cycle. Three dose levels of lenalidomide were evaluated (15, 20, 25 mg). Correlative studies included measurement of ADCC, FcγRIIIA polymorphism genotyping, measurement of serum cytokine levels, and flow cytometric analysis of immune cell subtypes. Twenty-two patients were enrolled (19 colorectal cancer, 3 HNSCC). Fatigue was the only dose-limiting toxicity. One partial response was observed and 8 patients had stable disease at least 12 weeks. The recommended phase II dose is cetuximab 500 mg/m2 with lenalidomide 25 mg daily, days 1–21. Correlative studies demonstrated a dose-dependent increase in natural killer cytotoxic activity with increasing doses of lenalidomide. Cetuximab and lenalidomide were well tolerated. There was a lenalidomide dose-dependent increase in ADCC with higher activity in patients enrolled in cohort 3 than those enrolled in cohorts 1/2. Although response was not a primary endpoint, there was evidence of antitumor activity for the combination therapy. Further investigation of lenalidomide as an immunomodulator in solid tumors is warranted. Mol Cancer Ther; 15(9); 2244–50. ©2016 AACR.
Abstract 1091: miR-125 family of miRNAs mediates prostate cancer cell proliferation and migration
Authors: Aherne Sinead T. , O’Neill Fiona , Madden Stephen F. , Clynes Martin , Lynch Conor C.
Published: July 2016

Background: Prostate cancer is the second most common malignancy in American men and causes ∼27540 deaths per annum. microRNAs (miRNAs) are a class of short non-coding RNAs that regulate the expression of protein-coding genes via either mRNA degradation or translational repression. These small molecules are known to be aberrantly expressed in prostate cancer (PCa) and possess both tumor suppressive and oncogenic properties. miR-125 is a highly conserved miRNA family comprised of three homolog members hsa-miR-125a, hsa-miR-125b-1, and hsa-miR-125b-2. Members of this family have been attributed both disease suppressing and promoting functions in certain contexts however their role in PCa has yet to be fully elucidated.

Methods: miRNA expression was evaluated in a publicly available dataset (GSE21032) comprised of 28 normal, 98 primary PCa and 13 metastasis samples. miR-125 expression was assessed in a panel of 8 prostate cell lines comprising PrEC prostate epithelial cells, RWPE-1, BPH1, DU145, LNCaP, C4-2B, PC3, and PC3-2M cell lines using the Qiagen miScript system. MirWalk was used to identify predicted targets of the miRNAs. miR-125 expression was transiently inhibited in LNCaP, C4-2B, PC3, and PC3-2M cell lines using miScript miRNA inhibitors and miRNA and target mRNA expression, cell proliferation and migration were measured post-transfection.

Results: Analysis of human data revealed miR-125a-3p was significantly up-regulated in metastasis samples compared to both primary PCa and normal tissues (log fold change (FC) 1.743, p-value 9.01E-08 logFC = 1.536 p-value = 8.80E-08 respectively). Next, we validated miR-125 family member expression in prostate cancer cell lines compared to PrEC and RWPE-1 prostate epithelial cells. Most notably, we observed that miR-125b-2 expression was significantly higher in all prostate cancer cell lines. Inhibition of miR-125b-2, significantly reduced the proliferation of the LNCaP, C4-2B, and PC3-2M cell lines. Moreover, inhibition of this miRNA significantly impaired prostate cancer cell migration by 43.8% (p-value = 0.003). Using MirWalk, we examined potential targets for regulation by miR-125b-2 and identified the potent tumor suppressor Quaking family as top hit. Our results show that inhibition of miR-125b-2 significantly enhanced the expression of Quaking family members (1.79 fold).

Conclusions: Our results demonstrate that the miR-125 family of miRNAs plays a role in regulating prostate cancer cell proliferation and migration, potentially by mitigating Quaking activity.

Citation Format: Sinead T. Aherne, Fiona O’Neill, Stephen F. Madden, Martin Clynes, Conor C. Lynch. miR-125 family of miRNAs mediates prostate cancer cell proliferation and migration. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1091.
Abstract 1133: Prevalence of activated NOTCH receptor in solid tumors and chronic lymphocytic leukemia
Authors: Oakley Gerard , Riegle Lisa , Benhadji Karim , Schade Andrew
Published: July 2016

Recent reports identified high expression of Notch 1 receptor in various tumors including adenoid cystic carcinomas (ACC). Notch 1 receptor is activated by gamma secretase cleavage to exert transcriptional control of processes including cellular differentiation and proliferation. Notch is also known to be activated in approximately 60% of T-cell acute lymphoblastic leukemias. We investigated the prevalence of Notch 1, 2 and 3 activation in different tumors by immunohistochemistry (IHC) designed separately for Notch 1, 2 and 3. Our proprietary antibodies recognize the post-gamma secretase cleavage activated Notch intracellular domain (NICD) fragments of Notch 1, Notch 2 and Notch 3, allowing direct measurement of activation of these receptors, in addition to their expression level by the tumor cells.

In total we tested 48 ACCs of the salivary glands, 19 breast cancer, 35 chronic lymphocytic leukemias (CLL), 7 glioblastomas, and 35 non-small cell lung carcinomas (NSCLC) We found that at least 65% (31 out of 48) of ACCs were positive for Notch 1 activation using our assay. Of other tumor types, 4 out of 7 glioblastomas (57%), 14 out of 35 CLLs (40%), 7 out of 35 NSCLC (20%) and 2 out of 19 breast cancer cases (11%) were positive for Notch 1. Within NSCLC, 5 of the 16 tested squamous cell carcinomas were positive for Notch 1 activation (31%). Notch 2 and Notch 3 showed only rare activation in these tumors. We conclude that high levels of Notch 1 activation are identifiable in different tumor types including high prevalence in ACC, CLL and glioma. These tumors may benefit from therapeutic Notch pathway inhibition.

Citation Format: Gerard Oakley, Lisa Riegle, Karim Benhadji, Andrew Schade. Prevalence of activated NOTCH receptor in solid tumors and chronic lymphocytic leukemia. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1133.
Abstract 1308: Enhanced antitumor activity of rapamycin and genipin, a UCP-2 inhibitor, in lung cancer
Authors: Su Wen-Pin , Chang Jang-Yang , Kuo Ching-Chuan , Su Wu-Chou
Published: July 2016

mTOR is constitutively activated in lung cancer. Deletion of TOR1 increases expression of OXPHOS proteins, and mitochondrial uncoupling proteins 2 (UCP-2) expression promotes chemoresistance. Therefore, we evaluated whether adding genipin, a UCP-2 inhibitor, to rapamycin has better antitumor effect in lung cancer. Combined with genipin and rapamycin have a synergistic effect with more cellular apoptosis in A549, H460 and CL1-0 cells. The similar phenomenon was found in the combination treatment with genipin and everolimus or BGT226 (a dual PI3K/mTOR inhibitor) in this three lung cancer cells. It is said that NrF2 may affect UCP-2 expression, and we found NrF2 was activated in lung cancer A549, H460 and CL1-0 cells treated with rapamycin. Everolimus and dual PI3K/mTOR inhibitor, BGT226, also induced NrF2 nuclear translocation in these lung cancer cells. It is said that under oxidant stress, p62 activates NrF2 through P62 Ser351-phosphorylation. We found that rapamycin induced ROS generation, p62 serine phosphorylation and NrF2 downstream mRNA and protein expression, including UCP-2. ROS inhibitor, NAC, and down-regulation of p62 by siRNA suppressed rapamycin-induced UCP-2 expression. Overexpression of p62 by cDNA transfection enhanced NrF2 activation and then UCP-2 expression in lung cancer cells. Moreover, rapamycin-induced increased UCP-2 expression, leading decreased mitochondrial membrane potential and mild increased oxygen consumption rate. Finally in A549 subcutaneous tumor mode in SCID mice, combined genipin and everolimus also demonstrated the greatest antitumor activity. Therefore, inhibition UCP-2 by genipin enhances anticancer effect of rapamycin in lung cancer.

Citation Format: Wen-Pin Su, Jang-Yang Chang, Ching-Chuan Kuo, Wu-Chou Su. Enhanced antitumor activity of rapamycin and genipin, a UCP-2 inhibitor, in lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1308.
Abstract 145: Comprehensive sequencing analyses of uterine and ovarian carcinosarcoma
Authors: Mori Seiichi , Gotoh Osamu , Sugiyama Yuko , Kato Kazuyoshi , Takazawa Yutaka , Noda Tetsuo
Published: July 2016

Carcinosarcoma (CS) of the uterus or the ovary, known as malignant mixed Muellerian tumor, is a biphasic neoplasm composed of malignant epithelial (carcinoma) and mesenchymal (sarcoma) elements. CS is a rare disease (two per 100,000 women) and exhibits poor prognosis (5-year survival rates: uterus and ovary; 31 and 7.5%, respectively). Treatment regimen for CS follows to that for endometrioid adenocarcinoma (EC) since CS has been considered as an aggressive form of high-grade EC. However, CS has a significantly worse prognosis than EC. Identification of a molecular target and/or a molecular mechanism in tumorigenesis from a large cohort study may improve unfavorable outcomes. Toward this aim, we perform targeted resequencing analysis with a panel of 596 genes, which were previously implicated as critical not only for uterine, ovarian or sarcomatus oncogenesis but also for molecular targets in therapeutics. Through analyses of 84 cases of uterine and ovarian CS, consensus clustering with deregulated pattern of CS genome identified four genomic subtypes that correlate with genetic or epigenetic alterations, histopathology and patient outcomes. Moreover, potentially actionable alterations were found in a substantial number of genes (including PIK3CA, MTOR, KRAS and CCNE1). This study would provide deep insights into the development of novel diagnostic and personalized treatment for CS patients.

Citation Format: Seiichi Mori, Osamu Gotoh, Yuko Sugiyama, Kazuyoshi Kato, Yutaka Takazawa, Tetsuo Noda. Comprehensive sequencing analyses of uterine and ovarian carcinosarcoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 145.
Abstract 1485: Sunitinib enhances the antitumor responses of agonistic CD40-antibody by reducing MDSCs and synergistically improving endothelial activation and T-cell recruitment
Authors: van Hooren Luuk , Georganaki Maria , Huang Hua , Mangsbo Sara , Dimberg Anna
Published: July 2016

CD40-activating immunotherapy has potent anti-tumor effects due to its ability to activate dendritic cells and induce cytotoxic T-cell responses. However, its efficacy is limited by accumulation of immunosuppressive cells in the tumor and by endothelial anergy induced by pro-angiogenic growth factors. Here, we show that combining agonistic CD40 monoclonal antibody (mAb) therapy with vascular targeting using the tyrosine kinase inhibitor sunitinib significantly decrease tumor growth and improve survival in B16.F10 melanoma and T241 fibrosarcoma. Treatment of tumor-bearing mice with anti-CD40 mAb lead to increased activation of CD11c+ dendritic cells in the tumor draining lymph node, as evidenced by enhanced CD86 expression, while sunitinib treatment reduced vessel density. CD11b+Gr1+ myeloid derived suppressor cells accumulated in the tumor draining lymph nodes after anti-CD40 mAb treatment. This effect was reversed by co-treatment with sunitinib. Tumor endothelial expression of ICAM1 and VCAM1 adhesion molecules were increased only when anti-CD40 mAb treatment was combined with sunitinib. This was associated with enhanced tumoral infiltration of CD8+ T-lymphocytes. Our results show that combining CD40-stimulating immunotherapy with sunitinib treatment exert potent complementary antitumor effects mediated by dendritic cell activation, reduction in myeloid derived suppressor cells and increased endothelial activation, resulting in enhanced recruitment of cytotoxic T cells.

Citation Format: Luuk van Hooren, Maria Georganaki, Hua Huang, Sara Mangsbo, Anna Dimberg. Sunitinib enhances the antitumor responses of agonistic CD40-antibody by reducing MDSCs and synergistically improving endothelial activation and T-cell recruitment. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1485.
Abstract 1638: Hypoxia-induced WSB1 promotes the metastatic potential of osteosarcoma cells
Authors: Wang Yijie , Cao Ji , Dong Rong , Lin Guanyu , Chen Zibo , Ding Ling , Ying Meidan , He Qiaojun , Yang Bo
Published: July 2016

Intratumoral hypoxia occurs in many solid tumors, where it is associated with the development of metastatic character. However, the connections between these phenomena are not fully understood. In this study, we define an integrative role for the E3 ubiquitin ligase subunit WSB1. In primary osteosarcomas, increased levels of WSB1 correlated with pulmonary metastatic potential. RNAi-mediated attenuation of WSB1 or disruption of its E3 ligase activity potently suppressed tumor metastasis. Quantitative proteomic and functional analyses revealed that WSB1 ubiquitylates the Rho-binding protein RhoGDI2 and promotes its proteasomal degradation, thereby activating Rac1 to stimulate tumor cell motility and invasion. Our findings show how WSB1 regulates key steps of the metastatic cascade in hypoxia-driven osteosarcoma, and highlight a candidate therapeutic target to potentially improve the survival of patients with metastatic disease.

Citation Format: Yijie Wang, Ji Cao, Rong Dong, Guanyu Lin, Zibo Chen, Ling Ding, Meidan Ying, Qiaojun He, Bo Yang. Hypoxia-induced WSB1 promotes the metastatic potential of osteosarcoma cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1638.
Abstract 1642: Calponin-3, acidic (CNN3) mediates EMT-related genes in human colorectal cancer metastasis
Authors: Kim Soon-Chan , Hong Chang-Won , Jang Sang-Geun , Kim Yeh-Ah , Jung Seung-Yong , Park Jea-Gab , Ku Ja-Lok
Published: July 2016

(a) An introductory sentence indicating the purposes of the study Distant metastasis is the major cause of cancer-related death in colorectal cancer (CRC) patients. Although the Calponin family has emerged as a distinguishing feature in epithelial-to-mesenchymal transition (EMT) in human cancer, the role of CNN3 member in the metastatic CRC has not been investigated.

(b) A brief description of pertinent experimental procedures. Three pairs of primary CRC and corresponding matched metastasis cell lines were analyzed for gene expression by microarray and western blotting. CNN3 was transduced to CRC cell line with lower CNN3 expression level (SW-480), and CNN3 with higher CNN3 expression CRC cell lines (SW-620, HCT-116) was suppressed by lentiviral-induced shRNA. Functional analysis of CNN3 overexpression and suppression was investigated in these CRC cell lines for proliferation and invasion. Expression of several potential CNN3 target genes (ERK1/2) and EMT markers (E-cadherin and vimentin) in primary CRC and matching metastasis CRC cell lines was validated.

(c) A summary of the new, unpublished data. Global CNN3 expression levels were significantly higher in metastasis cell lines compared with the matched primary cell lines in both mRNA (FC = 2.3, p = 0.017) and protein levels. CRC cell lines with suppressed CNN3 showed decreased cell proliferation and reduced invasive behavior in CRC cell lines. Suppression of CNN3 in CRC cell lines with higher CNN3 expression resulted in increased E-cadherin and decreased vimentin. pERK1/2 were decreased in accordance with suppression of CNN3 in both SW-620 and HCT-116. Overexpression of CNN3 in CRC cell lines caused reduced E-cadherin in SW-480 and HT-29.

(d) A statement of the conclusions CNN3 is involved in mediating EMT and metastatic behavior in the colon. Its expression is accordantly related to EMT markers, and CNN3 may serve as a possible diagnostic marker and therapeutic target for patients with CRC.

Citation Format: Soon-Chan Kim, Chang-Won Hong, Sang-Geun Jang, Yeh-Ah Kim, Seung-Yong Jung, Jea-Gab Park, Ja-Lok Ku. Calponin-3, acidic (CNN3) mediates EMT-related genes in human colorectal cancer metastasis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1642.
Abstract 1741: The association between menopause symptoms and risk of postmenopausal breast cancer among non-users of hormone therapy
Authors: Li Tricia , Han Jiali , He Chunyan
Published: July 2016

Background: Hot flashes are a common symptom among women during their menopause transition, likely due to the changes of endogenous hormone levels. Previous research suggested that hot flashes symptoms were associated with a reduced risk of cardiovascular diseases. Few studies have investigated the association of hot flashes symptoms with the risk of breast cancer. Furthermore, the confounding effect by hormone use has not been sufficiently addressed in previous studies.

Methods: We investigated the association between hot flashes symptoms and risk of postmenopausal breast cancer in the Nurses’ Health Study (NHS). In order to eliminate the confounding by hormone use, we investigated the association only among women who never used hormone therapy. We calculated relative risk of breast cancer using a multivariate Cox model, adjusting for potential confounders, including age, race, family history of breast cancer, age at menarche, age at menopause, parity, age at first birth, body mass index, smoking, alcohol use, and education. We further investigated the severity of hot flashes symptoms in association with risk of postmenopausal breast cancer.

Results: We documented 1079 incidence of breast cancer during 396,166 person-years of follow-up. Hot flashes symptoms were not associated with risk of postmenopausal breast cancer after adjustment for potential confounders (RR = 1.02, 95% CI, 0.90-1.15) among non-users of hormone therapy. Relative risk of breast cancer across symptom severity categories (mild, moderate, severe) were 1.01 (95% CI, 0.87-1.17), 1.04 (95% CI, 0.88-1.22), and 1.05 (95% CI, 0.75-1.46), respectively.

Conclusion: These data suggested that menopausal symptoms were not associated with risk of postmenopausal breast cancer.

Citation Format: Tricia Li, Jiali Han, Chunyan He. The association between menopause symptoms and risk of postmenopausal breast cancer among non-users of hormone therapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1741.
Abstract 188: Isoform-specific AKT functions in thyroid carcinoma development
Authors: Ranieri Michela , Di Cristofano Antonio
Published: July 2016

Mammalian cells contain three genes that encode three AKT isoforms (AKT1-3). The three genes have >85% sequence identity, and possess conserved threonine and serine residues that are critical for full activation. All AKT isoforms seem to have identical or similar substrate specificity, and it is not clear whether they possess different functional specificities in vivo.

Using conditional, organ-specific isoform deletion in a model of PI3K-dependent neoplastic transformation and progression, we are dissecting the molecular and biological roles of AKT1 and AKT2.

Thyroid-specific deletion of Pten (Ptenthy-/− mice) results in thyroid hyperplasia at birth which then slowly progresses to solid nodular lesions and finally to invasive carcinomas.

Simultaneous thyroid-specific deletion of Akt1 and/or Akt2 demonstrates that both AKT1 and AKT2 are necessary to confer proliferative ability to Pten−/- thyrocytes in an additive manner. However, with age, (Pten,Akt1)thy-/− mice have a significantly higher incidence of thyroid lesions compared to (Pten,Akt2)thy-/− mice. In addition, deletion of Akt2 is more effective than that of Akt1 to extend survival and reduce lesion aggressiveness in Ptenthy-/−/KrasG12D mice.

Transcriptomic and proteomic analyses have revealed that AKT2 loss rescues most of the expression and activation changes induced, in vivo, by Pten deletion, while AKT1 loss has more restricted effects.

Futhermore we have identified and are characterizing subsets of genes that are differentially regulated by AKT isoforms.

These data point to the existence of dramatically different molecular pathways associated with each AKT isoform in the thyroid, and represent a strong rationale for the development of less toxic, more effective treatments based on isoform-specific inhibitors.

Citation Format: Michela Ranieri, Antonio Di Cristofano. Isoform-specific AKT functions in thyroid carcinoma development. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 188.
Abstract 2082: Fc-FcγR interaction impacts the clearance and antitumor activity of antibody-drug conjugates in NSG mice
Authors: Li Fu , Ulrich Michelle , Hunter Joshua , Westendorf Lori , Olson Devra , Lee Cassie Baker , Benjamin Dennis , Law Che-Leung
Published: July 2016

Majority of preclinical efficacy studies for cancer therapeutics are performed on tumor models implanted in immune-deficient mouse strains, including athymic nude, severe combined immunodeficiency (SCID), or NOD-SCID/IL2Rgamma null (NSG) mice. To better understand the correlation between drug exposure and preclinical antitumor activity of biologics, we evaluated the pharmacokinetic (PK) profiles of antibodies and antibody-drug conjugates (ADC) in four strains of mice commonly used in cancer research, including BALB/c, athymic nude, SCID, and NSG. To our surprise, antibodies and ADCs had an abnormally short serum half-life in NSG mice, compared to other strains of mice.

We hypothesized that the fast clearance from plasma in NSG mice may reduce the overall exposure of xenografts to the ADCs, resulting in underestimation of antitumor activity. This was confirmed by comparing cAC10-vcMMAE in CD30+ Karpas-299 tumors implanted in either SCID mice or NSG mice. A single dose of 1 mg/kg cAC10-vcMMAE ADC resulted in complete remission of Karpas-299 tumors implanted in SCID mice (5/5). In contrast, the same treatment regimen had no effect on Karpas-299 tumors implanted in NSG mice (0/5). This observation suggests indeed the shortened exposure penalizes ADC efficacy in NSG models.

We then hypothesized that the fast clearance of ADCs from plasma in NSG mice could be mediated by Fc-FcγR interaction; compromising this interaction may improve the serum half-lives of antibodies and ADCs and thus enhance ADC-mediated antitumor effects in NSG mice. To test this, we utilized a mutant antibody with the amino acid substitutions of E233P:L234V:L235 in its Fc domain that result in significantly reduced FcγR binding (G1V1). These mutations restored the serum exposure in NSG mice to that level comparable to other strains of mice. As a result, treatment of 1 mg/kg cAC10G1V1-vcMMAE led to tumor remission in Karpas-299 tumors grown in NSG mice, correlating with extended exposure. In parallel, pretreatment with human IV immunoglobulin also enhanced ADC exposure and antitumor activity in the NSG mice.

In summary, these results suggest preclinical PK/PD assessment should be performed in the same strains of mice, especially when NSG xenograft models are used to evaluate activities. Moreover, reduced FcγR binding may enhance ADC exposure and allow a better assessment of therapeutic potential of biologics in NSG-based xenograft models.

Citation Format: Fu Li, Michelle Ulrich, Joshua Hunter, Lori Westendorf, Devra Olson, Cassie Baker Lee, Dennis Benjamin, Che-Leung Law. Fc-FcγR interaction impacts the clearance and antitumor activity of antibody-drug conjugates in NSG mice. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2082.
Abstract 2358: KHK2805, a novel ADCC- and CDC-enhanced anti-FOLR1 antibody with AccretaMab® technology, shows a potent anti-tumor activity in combination with pemetrexed
Authors: Ando Munetoshi , Nagata Keiko , Ishii Toshihiko , Nakai Ryuichiro , Takahashi Takeshi
Published: July 2016

Introduction: Folate receptor 1 (FOLR1, FR alpha) is a folate transporter which is expressed in many cancers including ovarian cancer (OvC) and non-small cell lung cancer (NSCLC), and which is an attractive target for cancer therapy, is currently the subject of ongoing studies. We established KHK2805, a novel anti-FOLR1 monoclonal antibody with AccretaMab® technology to enhance both the antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) activities. It was demonstrated that KHK2805 exhibits markedly high ADCC and CDC activity levels in clinical samples from ovarian cancer patients, while a tolerable safety profile has been observed in preclinical models using cynomolgus monkeys (100 mg/kg weekly for 4 weeks, intravenously). A greater understanding of the biology of FOLR1 is important for providing a novel therapeutic option for KHK2805 for patients with cancer. Pemetrexed (PEM), a second-generation anti-folate which inhibits thymidylate synthase, glycinamide ribonucleotide transformylase and dihydrofolate reductase, is used as a standard therapy for patients with cancers such as NSCLC. It is not fully understood how PEM treatment affects the expression of FOLR1 on cancer cells. We therefore examined the level of FOLR1 expression in cancer cells after PEM treatment, and the anti-tumor activity of KHK2805 in combination with PEM.

Materials and Methods: The FOLR1 expression levels after PEM treatment were examined in OvC, NSCLC, and endometrial cancer cells by flow cytometry. The ADCC activity of KHK2805 against PEM-treated cells was evaluated. The anti-tumor activity of KHK2805 in combination with PEM was investigated in SCID mice.

Results: Flow cytometry showed that PEM treatment increased the FOLR1 expression of various cancers such as OvC (IGROV1, SKOV3, MCAS), NSCLC (NCI-H1437, NCI-H2228), and endometrial cancer (MESSA, HEC1A, HEC1B) cells. In addition, PEM treatment enhanced the ADCC activity of KHK2805 against MCAS, NCI-H1437, and NCI-H2228, in comparison to cells that did not receive PEM treatment. Furthermore, the anti-tumor activity of KHK2805 in SCID mice bearing subcutaneous MCAS tumors was enhanced by PEM treatment.

Conclusions: PEM induced further FOLR1 expression in OvC, NSCLC, and endometrial cancer, resulting in the enhancement of the ADCC activity of KHK2805. The use of KHK2805 with PEM might therefore be a potent therapeutic option.

Citation Format: Munetoshi Ando, Keiko Nagata, Toshihiko Ishii, Ryuichiro Nakai, Takeshi Takahashi. KHK2805, a novel ADCC- and CDC-enhanced anti-FOLR1 antibody with AccretaMab® technology, shows a potent anti-tumor activity in combination with pemetrexed. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2358.
Abstract 2359: A-337, a potent bispecific antibody targeting EpCAM×CD3, as a potential immunotherapeutic agent for human solid tumors
Authors: Cui Yumin , Huang Zhihua , Chen Hanyang , Zhang Xinfeng , Qi Bo , Liu Heng , Yan Xiaoqiang
Published: July 2016

T cell bi-specific antibodies are potent molecules by binding and re-directing T cells to tumor cells through tumor associate antigens (TAAs), and triggering strong T cell activation and resulting in the killing of tumor cells. Bi-specific T cell engager antibody (BITE®) has demonstrated a remarkable clinical efficacy in liquid tumor. It remains to be a tremendous challenge for the development these types of bi-specific antibodies due to the induction of cytokine storm, thus limiting the dose-escalation in patients. A new immunotherapy antibody platform, (ITab TM), was developed to generate bi-specific antibody-like molecules. The molecules target TAA and activate human CD3 T cells simultaneously. A-337 is a bi-specific antibody targeting on EpCAM (Epithelial cell adhesion molecule) on tumor cells and CD3 in human T cells. In vitro, A-337 demonstrated high potency on activating human T cell measured by CD69 expression, and killing of EpCAM highly expressing tumor cell lines with EC50 values at < 100 pM in the presence of human peripheral blood mononuclear cells (PBMC). The human PBMC dependent cell killing activity of A-337 was correlated to the expression levels of EpCAM on the surface of tumor cell lines. In the mouse model, A-337 showed dose-dependent growth inhibition of SW480 xenograft. In cynomolgus monkey studies, A-337 by single IV infusion induced peripheral T cell re-distribution or activation and cytokine release and showed favorable pharmacokinetics features. The current studies reveal that A-337 has potent anti-tumor activities in vitro and in vivo. These results demonstrate that A-337 is a promising and potent immunotherapeutic agent to treat EpCAM-expressing solid tumors and warrants further development.

Citation Format: Yumin Cui, Zhihua Huang, Hanyang Chen, Xinfeng Zhang, Bo Qi, Heng Liu, Xiaoqiang Yan. A-337, a potent bispecific antibody targeting EpCAM×CD3, as a potential immunotherapeutic agent for human solid tumors. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2359.
Abstract 2376: Clonal evolution of skin carcinomas during tumor progression and metastasis
Authors: McCreery Melissa Q. , Balmain Allan
Published: July 2016

Tumor heterogeneity and the ongoing evolution of tumor subpopulations have presented considerable challenges to the success of cancer therapies, highlighting a critical need for greater understanding of these phenomena. We used a K5CreER-Confetti reporter mouse in combination with the DMBA (dimethylbenzanthracene)/TPA (12-O-tetradecanoylphorbol-13-acetate) skin carcinogenesis model to study the genetics, clonality, and evolutionary behavior of tumor subpopulations from tumor initiation to metastasis. Treating the skin of K5CreER-Confetti mice with tamoxifen results in widespread labeling of the epidermis and hair follicles, which persists for at least 18 months. Papillomas that develop from Confetti-activated skin are of clonal origin, evidenced by each tumor being distinctly dominated by a single Confetti color (RFP, YFP, GFP, or CFP; or uncolored). Twelve weeks after initiation, the majority of tumors remain single-colored, with only 22% (36 of 133) showing any evidence of a secondary color population present. By 20 weeks after initiation, however, nearly all tumors (78%, 42 of 54) have incorporated one or more secondary populations, readily identifiable as streaks of a distinct color. FACS separation and sequencing of these different colored populations reveals these populations are genetically distinct: in preliminary analysis, the primary color population typically carries the expected driver Hras Q61L mutation while the secondary populations do not. The tumor thus appears to be incorporating pseudo-normal tissue from its microenvironment during early growth. Activation of the Confetti reporter in tumors post-initiation in papillomas results in labeling of single cells with all 4 Confetti colors, allowing subpopulations and their evolution to be studied at later stages. These highly multi-color papillomas subsequently give rise to single-color carcinomas and metastases, suggesting that the conversion from a benign to malignant tumor is clonally driven. The Confetti mouse is a valuable tool for studying the clonality and genetics of subpopulations present in tumors, which need to be better understood in order to improve the treatment of heterogeneous human tumors.

Citation Format: Melissa Q. McCreery, Allan Balmain. Clonal evolution of skin carcinomas during tumor progression and metastasis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2376.
Abstract 2743: Finding determinants of PARP inhibitor sensitivity using genome-wide and focused CRISPR screens
Authors: Pettitt Stephen , Krastev Dragomir B. , Song Feifei , Ashworth Alan , Lord Christopher J.
Published: July 2016

We are using genomic approaches to study the mechanism of action of targeted cancer drugs, particularly poly(ADP-ribose) polymerase (PARP) inhibitors. One of these drugs, olaparib was recently approved for use in ovarian cancers with germline BRCA1/2 mutations and many other clinical trials with this and other PARP inhibitors are underway. Experimental study of potential resistance mechanisms can inform the ongoing clinical development of these drugs.

We previously carried out a loss of function mutagenesis screen in haploid cells, which showed that PARP1 was required for cellular toxicity of olaparib and other PARP inhibitors. This supported the hypothesis that PARP inhibitor toxicity occurs via a poisoning mechanism where inhibited PARP is tightly bound to DNA, forming a toxic lesion.

Recently we carried out a similar screen using genome-wide lentiviral CRISPR-Cas9 mutagenesis in diploid cells. Many PARP inhibitor resistant clones had loss of function Parp1 mutations as expected. However, we also isolated a point mutation affecting a single amino acid in the Parp1 DNA binding domain. This mutant encodes a stable Parp1 protein that cannot bind DNA and does not become trapped in the presence of inhibitors. Thus the CRISPR screen implicated PARP1 DNA binding directly in determination of PARP inhibitor toxicity and was extremely informative about the mechanism of action compared to conventional loss-of-function mutagenesis.

We extended this approach by synthesising a high-density focused sgRNA library targeting only PARP1. We developed a reporter cell line that allows us to selectively isolate in-frame mutations that preserve PARP1 protein expression. By deep sequencing mutagenised and appropriately selected cells we identified a series of subtle mutations in PARP1 that result in PARP inhibitor resistance, giving us a detailed insight into structure-function relationships in PARP1. The approach is applicable to other drug targets, providing a framework for understanding drug action and predicting clinical resistance.

Citation Format: Stephen Pettitt, Dragomir B. Krastev, Feifei Song, Alan Ashworth, Christopher J. Lord. Finding determinants of PARP inhibitor sensitivity using genome-wide and focused CRISPR screens. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2743.
Abstract 290: NF1 loss modulates cellular fitness in KIT-mutant gastrointestinal stromal tumor (GIST)
Authors: Marino-Enriquez Adrian , Li Bin , Bahri Nacef , Lauria Alexandra , Wang Yuexiang , Fletcher Jonathan A.
Published: July 2016

Patients with inoperable GIST obtain substantial clinical benefit from imatinib and other KIT/PDGFRA inhibitors. However, secondary resistance develops in 90% of patients within 18-24 months. Reactivation of oncogenic signaling pathways at time of progression is most often due to secondary mutation in KIT or PDGFRA, but downstream RAS or BRAF mutations are occasionally alternative mechanisms of KIT/PDGFRA-inhibitor resistance.

We identified deleterious NF1 mutations in 3 KIT-mutant GIST samples from patients progressing on imatinib. In genomic scale loss-of-function shRNA genomic screens, designed to identify genes involved in imatinib resistance, a shRNA targeting NF1 was the most enriched shRNA in imatinib-treated GIST cells out of 54,000 shRNAs (NF1 gene score 29/11,238). To evaluate the contribution of NF1 loss to KIT-mutant GIST resistance, NF1-deficient GIST models were generated from 2 patient-derived, KIT-mutant GIST cell lines, GIST882 and GIST-T1, by sequential cotransfection with Cas9 and 2 independent NF1-targeting CRISPR-sgRNAs.

Complete loss of NF1 protein was observed after 5-10 passages due to on-target heterogeneous genomic deletion at the NF1 locus. Orthogonal validations were obtained using stable shRNA-mediated NF1 knockdown with similar results in the same GIST cell lines. NF1-deleted cells retained expression of mutant KIT but were resistant to imatinib (IC50 196 and 500nM, significantly increased from baseline 34 and 110nM, for GIST-T1 and GIST882, respectively). Notably, after interruption of imatinib treatment, NF1-deficient GIST cells showed reduced proliferation (69% and 77% growth reduction at day 7 off imatinib, for GIST-T1 and GIST882, t test p<0.001) despite continuous expression of activated KIT and downstream signaling effectors.

In conclusion, acquired NF1 loss induces TKI resistance in KIT-mutant GIST cells but results in reduced cellular fitness in the absence of KIT-inhibitor treatment. These findings suggest that NF1-deficient GIST subclones that expand on imatinib treatment can be destabilized by intermittently discontinuing KIT-inhibitor therapy.

Citation Format: Adrian Marino-Enriquez, Bin Li, Nacef Bahri, Alexandra Lauria, Yuexiang Wang, Jonathan A. Fletcher. NF1 loss modulates cellular fitness in KIT-mutant gastrointestinal stromal tumor (GIST). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 290.