bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2938: Transmembrane chloride intracellular channel 1 (tmCLIC1) correlates with metastatic potential of colorectal cancer cells
Authors: Carlini Valentina , Cianci Francesca , Verduci Ivan , Mazzanti Michele
Published: August 2019

Colorectal cancer (CRC) is the third most common and lethal tumor worldwide. Although CRC can be efficiently eradicated when diagnosed in the early stage, no effective therapies are currently available for advanced metastatic disease. Therefore, targeting metastasis represents a critical challenge for the successful treatment of CRC. To achieve this purpose, our laboratory has found a promising candidate inthe transmembrane localized Chloride Intracellular Channel 1 (tmCLIC1) protein. Growing scientific evidence have suggested an involvement of tmCLIC1 in different human malignancies, including CRC, with an unknown mechanism. CLIC1 is a metamorphic protein able to shuttle between a soluble cytoplasmic and a membrane integrated form, the latter characterized by ionic channel function. In normal cells, it is mostly cytoplasmic and cooperates to the maintenance of several physiological processes. Otherwise, in cancer cells CLIC1 undergoes to chronic membrane insertion, where contributes to the abnormal cellular hyperproliferative rate.In the present investigation, using CRC human cell lines at different stage of tumor development, we demonstrate that tmCLIC1 functional expression correlates with CRC cells increasing aggressiveness, while it is almost absent in normal colon cells. We also show that CLIC1 enrichment in plasma membrane is involved in promotion of CRC metastasis. CLIC1 inhibition using different pharmacological treatments led to drastic reduction of cell proliferation, migration and invasiveness.tmCLIC1 function permits to discriminate between normal and malignant cells and its downregulation is able to arrest metastatic cells. This is not the case of other upregulated proteins not only involved in tumorigenesis but also in physiological cell functions. Since metastasis are considered the leading cause of treatment failure and tumor relapse, CLIC1 inhibition coupled to conventional therapies can represent a novel strategy to successfully treat CRC with reduced side effects. (This work is supported by a GRANT from AIRC to MM)

Citation Format: Valentina Carlini, Francesca Cianci, Ivan Verduci, Michele Mazzanti. Transmembrane chloride intracellular channel 1 (tmCLIC1) correlates with metastatic potential of colorectal cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2938.
Abstract 2949: The impact of drug perturbations on neuroblastoma disease pathways: A pharmaco-transcriptomics approach
Authors: Elgendy Ramy , Almstedt Elin , Vanlandewijck Michael , Nelander Sven
Published: August 2019

Neuroblastoma (NB) cells exhibit a complex spectrum of pathway changes associated with oncogene activation, chromosome events, tumor micro-environment and super-enhancer states. So far, elucidating which pharmaceutical compounds could modulate the activation level of each known pathway in NB cells has not been feasible. To solve this problem, we have combined transcriptome profiling of drug-perturbed NB cells with mathematical modeling of public data to create a first map of drug- and NB-specific transcriptional signatures for more than 7000 pharmaceutical compounds. We treated 2 patient-derived xenograft (PDX) NB cell lines with different chemical compounds, at 3 different doses (IC50, IC20, and IC10) and 2 time-points (6 and 24h). The whole-transcriptome of the treated cells was analyzed by a cost-effective hybrid ‘PLATE-Seq/SMART-Seq2’ method. Using factor analysis and supervised machine learning, the data was used to build a model that predicts the NB-specific outcome of 7035 drugs studied in other (non-NB) cell lines in the L1000/LINCS project. Analysis of 768 NB-specific transcriptome profiles, showed that our method accurately detects dose-dependent drug-induced pathway changes in the NB cells, including alteration of characteristic pathways and risk signatures. From the 768 profiles, our best-fitted machine learning model was then extended to 7035 compounds by in silico extrapolation, providing a first panel of estimated drug effects in NB cells. Our constructed NB pharmacological map provides a detailed view of both predicted and in vitro-validated drug-NB interactions. Effective treatments for high-risk NB are currently lacking. Using a new and cost-effective strategy, we could build an accurate map of drug-induced pathway alterations, for network pharmacology and drug repositioning for NB patients.

Citation Format: Ramy Elgendy, Elin Almstedt, Michael Vanlandewijck, Sven Nelander. The impact of drug perturbations on neuroblastoma disease pathways: A pharmaco-transcriptomics approach [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2949.
Abstract 2951: A genome-wide association study identifies five novel genetic markers for trastuzumab-induced cardiotoxicity
Authors: Udagawa Chihiro , Hara Mari , Shimo Arata , Kojima Yasuyuki , Yoshie Reiko , Zaha Hisamitsu , Abe Norie , Motonari Tokiwa , Unesoko Mikiko , Tamura Kenji , Shimoi Tatsunori , Yoshida Masayuki , Yoshida Teruhiko , Okamura Hiroshi , Mushiroda Taisei , Tsugawa Koichiro , Zembutsu Hitoshi
Published: August 2019

Purpose: Trastuzumab has been administered to HER2-positive breast or gastric cancer patients as an effective treatment, however, the cardiotoxicity is identified as one of the life-threatening side effects. To identify a novel genetic marker(s) determining the risk of trastuzumab-induced cardiotoxicity, we performed a genome-wide association study (GWAS).

Experimental Design: We carried out a GWAS using 11 cases (with trastuzumab-induced cardiotoxicity) and 257 controls (showing no sign of trastuzumab-induced cardiotoxicity). Top 100 single nucleotide polymorphisms (SNPs) which revealed smallest P value in GWAS (P = 1.61 x 10-7 -1.97 x 10-4) were further examined using replication samples consisted of 14 cases and 199 controls. Results: The combined analysis of the GWAS and replication study indicated possible association of five loci with trastuzumab-induced cardiotoxicity (chromosome 13q14.3, 4q25, 15q26.3, 17q25.3 and another 15q26.3, Pcombined = 6.00 x 10-6, 8.60 x 10-5, 1.01 x 10-4, 1.07 x 10-4, 1.60 x 10-4, respectively). Furthermore, we developed a risk prediction model for trastuzumab-induced cardiotoxicity, using five SNPs which revealed smallest P value at each locus. The incidence of trastuzumab-induced cardiotoxicity in patients with risk score ≥5 was 42.5% (P =8.69 x 10-15) . This scoring system using five candidate loci could be used to predict the risk of the adverse event before administration of trastuzumab.

Conclusion: We identified five novel loci, which could be associated with trastuzumab-induced cardiotoxicity. These findings provide new insights into personalized trastuzumab therapy for patients with HER2-positive cancer.

Citation Format: Chihiro Udagawa, Mari Hara, Arata Shimo, Yasuyuki Kojima, Reiko Yoshie, Hisamitsu Zaha, Norie Abe, Tokiwa Motonari, Mikiko Unesoko, Kenji Tamura, Tatsunori Shimoi, Masayuki Yoshida, Teruhiko Yoshida, Hiroshi Okamura, Taisei Mushiroda, Koichiro Tsugawa, Hitoshi Zembutsu. A genome-wide association study identifies five novel genetic markers for trastuzumab-induced cardiotoxicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2951.
Abstract 2956: Inhibition of CHK1 sensitizes Ewing sarcoma cells to CDK1-dependent cell death in S-phase
Authors: Gordon David , Goss Kelli , Koppenhafer Stacia
Published: August 2019

Ewing sarcoma is a highly aggressive bone and soft tissue cancer that is caused by the EWS-FLI1 fusion protein. The treatment of Ewing sarcoma has changed very little in the past two decades and novel treatment approaches are needed. We recently identified that Ewing sarcoma cells are uniquely vulnerable to inhibitors of ribonucleotide reductase (RNR), the rate limiting enzyme in the synthesis of deoxyribonucleotides. We subsequently found that the inhibition of checkpoint kinase 1 (CHK1) increases the sensitivity of Ewing sarcoma cells to inhibitors of RNR, such as gemcitabine. The ATR-CHK1 pathway, from a mechanistic standpoint, regulates multiple stages of the cell cycle, including S-phase, the G2/M transition, and M phase. In the current work, we used cell synchronization to identify that the inhibition of CHK1 in S-phase cells results in premature entry into mitosis and cell death. Similarly, the inhibition of ATR serine/threonine kinase (ATR), the upstream activator of CHK1, also causes Ewing sarcoma cells to inappropriately enter mitosis. Moreover, we have found in Ewing sarcoma cells that this aberrant entry into mitosis is mediated, in part, by cyclin dependent kinase 1 (CDK1). In addition, activation of CDK1 by inhibiting the WEE1 kinase with AZD1775 also results in entry into mitosis and cell death in Ewing sarcoma cells. Currently, ongoing work is focused on the in vivo testing of gemcitabine in combination with CHK1 and WEE1 inhibitors as a novel therapeutic approach for the treatment of Ewing sarcoma.

Citation Format: David Gordon, Kelli Goss, Stacia Koppenhafer. Inhibition of CHK1 sensitizes Ewing sarcoma cells to CDK1-dependent cell death in S-phase [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2956.
Abstract 2957: Nasopharyngeal cancer
Authors: Kam Ngar Woon , Chan See Wing , Tsao George Sai-Wah , Guan Xin-Yuan , Kwong Dora Lai Wan
Published: August 2019

Background:

Nasopharyngeal cancer (NPC) is associated with a poor survival rate. The ability of cancer cells to evade apoptosis and exhibit limitless replication potential allows for the progression of cancer. The aim of this study was to investigate in vitro the effect of the isoflavone phenoxodiol on human NPC cell line and T cells, which has previously been shown to inhibit cell proliferation and promote apoptosis in a range of cancer cell lines and in anti-CD3/anti-CD28-activated murine splenocytes.

Methods:

Four NPC cancer cell lines-C666, C17, and paired NPC43 EBV-positive and its respective EBV-negative cell lines were cultured in vitro, and then treated with phenoxodiol for 72h or 5 days. The migration and proliferation was investigated by transwell and XTT, respectively. Conditioned medium of treated NPC cell lines were collected and incubated with human PBMC for 5 days. CD4 and CD8 surface staining were analyzed by flow cytometry.

Result:

We demonstrated that phenoxodiol inhibited NPC migration and cell proliferation (IC50 4µM). Among all the NPC cell lines, the proliferation effect on EBV-negative cell line (NPC43-ve) is least responsive to phenoxodiol. A dose-dependent of apoptosis in NPC cell lines by phenoxodiol [early apoptotic cells (Annexin V+/PI- at 1µM); late apoptotic cells (Annexin V+/PI+ at 2µM), and necrotic cells (Annexin V-/PI+, at 4µM) were distinguished. Interestingly, phenoxodiol reduced frequency of CD4 T cells while promoting the number of CD8 T cells.

Conclusions:

Phenoxodiol demonstrates an ability in NPC cancer cells to induce its apoptosis while inhibit its migration and proliferation. Moreover, the ability of phenoxodiol to modulate T cells population indicates that phenoxodiol would be effective as a potential future treatment for NPC.

Note: This abstract was not presented at the meeting.

Citation Format: Ngar Woon Kam, See Wing Chan, George Sai-Wah Tsao, Xin-Yuan Guan, Dora Lai Wan Kwong. Nasopharyngeal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2957.
Abstract 2961: Ganoderma lucidum extracts inhibit mTORC1/2 by activating AMPK and inhibiting IGFR/PI3K/Rheb in tumor cells
Authors: Huang Shile , Sohretoglu Didem , Zhang Chao , Luo Jun
Published: August 2019

Ganoderma lucidum (G. lucidum) extracts, as dietary supplements, have been found to possess potent anticancer activity, which is attributed to the presence of polysaccharides and triterpenes. However, the molecular mechanism underlying the anticancer action of G. lucidum extracts remains to be elucidated. Here we show that ReishiMax GLp, containing both polysaccharides and triterpenes of G. lucidum (GLPT), inhibited cell proliferation and induced cell death in human lung cancer cells (A549 and A427), and concurrently suppressed the signaling pathways mediated by the mammalian target of rapamycin (mTOR), a central regulator of cell proliferation and survival. Interestingly, GLPT not only inhibited mTORC1-mediated phosphorylation of p70 S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E binding protein 1 (4E-BP1), but also repressed mTORC2-mediated phosphorylation of Akt. Mechanistically, GLPT downregulated the phosphorylation and protein levels of insulin-like growth factor 1 receptor (IGFR) and phosphoinositide 3-kinase (PI3K), as well as the protein level of RAS homolog enriched in brain (Rheb). Besides, GLPT also activated AMP-activated protein kinase (AMPK) network. This is supported by the findings that GLPT increased the phosphorylation of AMPKα (T172), as well as its substrates tuberous sclerosis complex 2 (TSC2, S1387) and regulatory-associated protein of mTOR (raptor, S792). Ectopic expression of dominant negative AMPKα partially attenuated the inhibitory effect of GLPT on mTORC1, indicating that GLPT inhibits mTORC1 partly via activating AMPK. The results suggest that G. lucidum extracts execute the anticancer action at least partly by inhibiting the mTORC1/2 signaling through activation of AMPK and inhibition of IGFR/PI3K/Rheb in tumor cells. Supported by the Feist-Weiller Cancer Center, LSU Health Sciences Center, Shreveport, LA, USA.

Citation Format: Shile Huang, Didem Sohretoglu, Chao Zhang, Jun Luo. Ganoderma lucidum extracts inhibit mTORC1/2 by activating AMPK and inhibiting IGFR/PI3K/Rheb in tumor cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2961.
Abstract 2962: Targeting BCL2 as a therapeutic strategy in neuroendocrine prostate cancer
Authors: Corella Alexandra N. , Lucas Jared M. , Kaipainen Arja , Coleman Ilsa M. , Morrissey Colm , Corey Eva , Nghiem Paul , MacPherson David , Nelson Peter S.
Published: August 2019

Small-cell neuroendocrine prostate carcinoma (SC/NEPC) is an aggressive subtype of prostate cancer (PC) with poor response to conventional therapies and no approved targeted therapies. The molecular mechanisms that drive the growth and survival of these tumors are poorly understood due to disease rarity and a lack of model systems for the subtype. SC/NEPC shares defining characteristics with other aggressive small-cell neuroendocrine tumors, such as “small-blue round cell” morphology and expression of chromogranin A. In this study, we compare whole transcriptome RNA-sequencing data from metastatic PCs and two additional SC/NE tumor types, small cell lung cancer (SCLC) and Merkel Cell carcinoma (MCC). By differential expression analysis of each neuroendocrine tumor type to Androgen Receptor-responsive Prostate Cancer (AR-PC), we identify 4,300 genes exhibiting shared expression patterns in multiple neuroendocrine tumor types. Among these “pan-neuroendocrine” genes are known and proposed drivers of the neuroendocrine transcriptional program including MYCN (p=8.7E-07, upregulated in SC/NEPC over AR-PC) and potential novel drivers of the neuroendocrine phenotype in PC such as MYCL (p=3.5E-05, upregulated in SC/NEPC over AR-PC). We also identify conserved expression patterns of druggable targets including BCL2 which has previously been reported as highly expressed in subsets of SCLC and MCC, but not SC/NEPC (p=1.24E-05, upregulated in SC/NEPC over AR-PC). We then analyzed a panel of cell lines and novel PDX models, which further confirm that the BCL2 protein is highly expressed specifically in the SC/NEPC phenotype. We determined that SC/NEPC cell lines and PDX models are sensitive to BCL2 family member inhibitors targeting BCL2, BCLXL, BCLW, including, ABT263, ABT199 and ABT737, at sub-micromolar concentrations, whereas models of ARPC were resistant. These inhibitors lead to apoptotic cell death in SC/NEPC. Collectively, our data suggests that BCL2 inhibition may present a novel targeted approach for SC/NEPC that warrants further evaluation.

Citation Format: Alexandra N. Corella, Jared M. Lucas, Arja Kaipainen, Ilsa M. Coleman, Colm Morrissey, Eva Corey, Paul Nghiem, David MacPherson, Peter S. Nelson. Targeting BCL2 as a therapeutic strategy in neuroendocrine prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2962.
Abstract 2969: The antiviral agent Cidofovir induces DNA damage and mitotic catastrophe in HPV-positive and -negative head and neck squamous cell carcinomas in vitro
Authors: Speel Ernst-Jan M. , Verhees Femke , Legemaate Dion , Jacobs Robin , Haakma Wisse E. , Rousch Mat , Kremer Bernd
Published: August 2019

Objectives: Cidofovir (CDV) is an antiviral agent with anti-proliferative properties and is suggested as treatment option in head and neck squamous cell carcinoma (HNSCC) patients to improve outcome and quality of life. The mechanisms underlying the effectivity of CDV are not completely understood. The aim of our study was to investigate the efficacy of CDV in HPV-positive and -negative HNSCC cell lines in vitro and whether it is caused by a difference in response to DNA damage.

Materials and methods: Upon CDV treatment of HPV-positive and -negative HNSCC-, uterine cervical carcinoma (UCC)- and immortalized NOK- cell lines, cell viability was assessed with a MTT assay. Accumulation of DNA double strand breaks (DSBs) and activation of the DNA repair pathway were analyzed using immunofluorescence (IF) and Western blotting (WB), respectively. Apoptosis was detected by cleaved-PARP (WB) and mitotic catastrophe by phospho-Aurora Kinase and Cyclin B1 (IF).

Results: All cell lines responded to CDV. Treatment resulted in γ-H2AX accumulation and upregulation of DNA repair proteins, especially in the HPV-positive cells. CDV did not induce PARP cleavage but induced Cyclin B1 expression. Phospho-Aurora Kinase immunostaining showed a decrease in number of mitoses but an increase in aberrant mitoses suggesting mitotic catastrophe upon CDV treatment.

Conclusion: CDV inhibits cell growth in HPV-positive and -negative HNSCC and UCC cell lines which was more profound in HPV-positive cell lines. CDV treated cells showed accumulation of DNA DSBs. Although the DNA repair was activated, apoptosis did not occur. Rather our data indicate the occurrence of mitotic catastrophe.

Citation Format: Ernst-Jan M. Speel, Femke Verhees, Dion Legemaate, Robin Jacobs, Wisse E. Haakma, Mat Rousch, Bernd Kremer. The antiviral agent Cidofovir induces DNA damage and mitotic catastrophe in HPV-positive and -negative head and neck squamous cell carcinomas in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2969.
Abstract 2999: Hybrid complex between bispecific peptide and antibody for targeted cancer therapy
Authors: Yu Byeongjun , Hwang Dobeen , Jon Sangyong , Chung Junho
Published: August 2019

Antibodies have been used for cancer therapy for two decades, but they suffer from poor localization and penetration into solid tumors because of their large size. Peptides can be an alternative for treatment of solid tumors due to their small size. However, therapeutic use of peptides is limited because of their short plasma half-life. We present a new molecular platform based on a hybrid complex between bispecific peptide and antibody, designated as HyPEP-body, composed of bispecific peptides labeled with a hapten as a pharmacophore and an anti-hapten antibody. Two different peptides are joined together with linker and then, labeled with the hapten. The bispecific pharmacophore are bound to the anti-hapten antibody through antigen-antibody interaction. The pharmacophores which can be chemically optimized or replaced with other peptides are in charge of therapeutic function, while the antibody imparts long plasma half-life. As a model study, we constructed HyPEP-body that simultaneously binds extra domain B (EDB) and vascular endothelial growth factor (VEGF) with EDB binding peptide, VEGF binding peptide, cotinine and anti-cotinine antibody. Complexation with the antibody extended the plasma half-life of the bispecific pharmacophores. Furthermore, bispecific pharmacophores released from the antibodies distributed more widely in the tumor tissue than the antibodies. Finally, treatment of the HyPEP-body inhibited tumor growth in U87MG, human glioblastoma, xenograft mouse model. This versatile hybrid platform will enhance the value of peptide therapeutics and suggest an innovative solution to the development of bispecific antibodies that has suffered from productivity issues.

Citation Format: Byeongjun Yu, Dobeen Hwang, Sangyong Jon, Junho Chung. Hybrid complex between bispecific peptide and antibody for targeted cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2999.
Abstract 30: The development of a 3D cell-culture model of drug-induced oral mucositis
Authors: Lambros Maria P. , Fei QinQin , Moreno Jonathan
Published: August 2019

Purpose: Mucositis is a debilitating disease affecting cancer patients. Mucositis is an adverse effect of radiation or chemotherapy. In the past, our lab has developed a model of radiation-caused mucositis using a validated human tissue substitute of oral mucosa and studied the post radiation cellular and molecular stresses and their prevention. In this project, we wished to develop an in-vitro model of chemical-, or drug-induced oral mucositis. For this purpose, we used a three dimensional (3-D) cell culture model of oral mucosa and as the drug to induce mucositis we used everolimus. Everolimus belongs to rapalogs which represent important therapies against cancer but one of its adverse-effects is mucositis. Mucositis caused by everolimus develops early in therapy and progresses quickly to grade 3 making patients to postpone life-saving therapies. Thus, it is very important to understand and prevent the development of everolimus-caused mucositis.

Methods: The 3-D cell cultures of primary human oral keratinocytes grown on top of primary human oral fibroblasts were purchased from MatTek Corporation. These 3-D cell cultures (tissues) are similar to human oral mucosal tissue and have been validated as its substitutes. Different 3-D tissues were treated for 24, 40 and 60 hours with 8, 16, 32, and 64 ng/ml of everolimus. Tissues which received no treatment were used as controls. We then evaluated the tissue histology using H and E staining, and gene expression using RNA-sequencing.

Results: Tissues which were treated with everolimus show signs of distress based on the concentration of everolimus and the duration of the treatment. Low and medium concentration of everolimus (8ng/ml) and (16 or 32 ng/ml), respectively, show no, or minimal tissue distress after 24 hours of treatment duration. Tissues which received a high concentration of everolimus (64 ng/ml) even at a short duration of treatment, show signs of distress, as evidenced by atypical proliferation at the keratinous epithelium, vacuoles, or shredded keratin layers, as well as, higher apoptosis, and expression of pro-inflammatory cytokine genes. In general, the higher the concentration of everolimus and the longer the duration of the treatment, the more the signs of the stress in tissues.

Conclusion: The cell and molecular stress that the 3-D tissue cultures of human oral mucosa experience depends on the drug concentration and the duration of treatment.

Acknowledgment We thank Novartis for their support.

Citation Format: Maria P. Lambros, QinQin Fei, Jonathan Moreno. The development of a 3D cell-culture model of drug-induced oral mucositis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 30.
Abstract 3008: PSMA glycosylation and aggressive prostate cancer progression
Authors: Burch Tanya C. , Mackay Stephen S. , Oduor Ian O. , Otto Joseph J. , Lance Raymond S. , Troyer Dean A. , Semmes Oliver John , Nyalwidhe Julius O.
Published: August 2019

Introduction: Prostate cancer has a heterogeneous spectrum of clinical outcomes with phenotypes ranging from indolent asymptomatic cases to very aggressive metastatic and lethal forms. Our long term objective is to identify molecules and characterize mechanisms for PCa progression to aggressive lethal disease. PSMA, a membrane bound glycoprotein is significantly over-expressed in more aggressive and metastatic prostate carcinomas with a negative correlation with patient prognosis. The role of its glycans in disease progression and in the development of biomarkers for the disease and potential therapeutics are currently unknown. Aberrant glycosylation of glycoproteins has been observed in many malignancies, and these changes influence disease progression. Our rationale is that differential glycosylation of PSMA correlates with PCa aggressive disease progression. In this study, we characterize N-linked glycosylation profiles in disease stratified PCa cells and prostate proximal fluids show differential expression of specific PSMA conjugated glycan structures that correlates with aggressive phenotypes.

Methods/Results: PSMA was isolation from PCa cell lysates and post-DRE urines by column based affinity chromatography followed by SDS-PAGE and Western Blot and Coomassie staining. The fractionated PSMA protein band was excised and processed for proteomic and glycomic analysis. Isolated PSMA was trypsin digested and treated with PNGaseF and other glycosidases to cleave of N-linked glycans from the peptides. Isolated glycans were subjected to permethylation before MALDI-TOF-MS and high resolution, high mass accuracy and high sensitivity LC/MS/MS on an Orbitrap Fusion Lumos Tribrid MS instrument with multiple fragmentation capabilities (CID, HCD, ETD and EThcD) for glycan identification and quantitation. Validation of differential glycan expression was further performed by targeted glycan based MS and lectin based biochemical technologies.

Conclusions: Our preliminary studies show that there is differential expression of PSMA N-linked glycans with disease severity in prostate cancer cells and post DRE urine samples. Further analyses are currently underway using an extensive cohort of post DRE urine samples and additional studies are focusing on uncovering the mechanisms that are modulated by PSMA glycosylation that are responsible for PCa disease progression. These could assist in the development of novel diagnostic and therapeutic strategies for the disease.

Citation Format: Tanya C. Burch, Stephen S. Mackay, Ian O. Oduor, Joseph J. Otto, Raymond S. Lance, Dean A. Troyer, Oliver John Semmes, Julius O. Nyalwidhe. PSMA glycosylation and aggressive prostate cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3008.
Abstract 3021: The role of survivin splice variants in pancreatic ductal adenocarcinoma Gemcitabine resistance
Authors: Kabagwira Janviere , Gonda Amber , Kwong Mei Li
Published: August 2019

Pancreatic Ductal Adenocarcinoma (PDAC) is a highly lethal disease with poor prognosis and an increased chemoresistance. Dysfunction in alternative splicing has been associated with chemoresistance in many cancers. Alternative splicing of survivin mRNA generates six splicing isoforms some of which are associated with cancer progression and chemoresistance. Survivin is a member of the inhibitors of apoptosis (IAPs) family and a cell cycle-associated oncoprotein overexpressed in almost all cancers. Interestingly, some of these survivin splice variants are proapoptotic (2α and 2β) while others are antiapoptotic (survivin, Δex3, 3α and 3β). It has been reported that increased levels of survivin 2β and 3β play a role in chemoresistance in ovarian cancers and other tumors, but this has not been studied in pancreatic cancer. Our recent RT-qPCR data also show an overexpression of all survivin splice variants in chemoresistant PDAC cell lines compared to the chemo-sensitive cell line. We, therefore, hypothesize that increased expression of survivin splice variants 2β and 3β contribute to chemo-resistance in PDAC. To test this hypothesis, we repeatedly selected the resistant cells from the normally chemosensitive MiaPaca2 cell line after exposure to gemcitabine. We then assessed the levels of survivin splice variants in the new cell line (gemcitabine-resistant MiaPaca2). Using the Alamar blue assay, we confirmed that Panc1 and gemcitabine-resistant MiaPaca2 are more resistant to gemcitabine compared to MiaPaca2 cells. We also observed a statistically significant overexpression of survivin 2β and 3β in gemcitabine-resistant cell lines compared to the more sensitive MiaPaca2 cells. To further test our hypothesis, we will knockdown survivin 2β and 3β in Panc1 and gemcitabine-resistant MiaPaca2, and/or overexpress these splice variants in MiaPaca2 cells then assess response to gemcitabine. The outcomes of this study might create new biomarkers for predicting the clinical outcomes of gemcitabine treatment and generate new therapeutic targets for pancreatic cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Janviere Kabagwira, Amber Gonda, Mei Li Kwong. The role of survivin splice variants in pancreatic ductal adenocarcinoma Gemcitabine resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3021.
Abstract 3041: CD72 as a potential new therapeutic target in diffuse large B-cell lymphoma (DLBCL) and mantle cell lymphoma (MCL)
Authors: Euw Erika Von , Taschereau Eileen , Ou Da Ming , Mead Monica , Larson Sarah , Slamon Dennis
Published: August 2019

Cluster of differentiation 72 (CD72) is a 45 kDa type II transmembrane glycoprotein expressed primarily on the surface of B cells. In a search for a novel lymphoma specific target, public datasets were leveraged to assess mRNA expression. We investigated CD72 mRNA expression in cell lines (data from CCLE), patients (data from TCGA) and normal tissues (data from GTEx). The absence of CD72 in normal tissue and the presence in B cells and non-Hodgkin lymphomas (B-NHL) made CD72 a potential target for the treatment of B-NHL, mainly DLBCL and MCL.

CD72 expression on the cell surface was tested by FACS in 15 DLBCL and 5 MCL cell lines. Of these, 12 DLBCL and 4 MCL were positive for CD72, while 3 DLBCL and 1 MCL showed no CD72 expression. In all cell lines tested, the cell surface expression of CD72 correlated with mRNA expression.

To evaluate the potential of CD72 as an ADC target, 4 cell lines were treated in vitro with three commercial monoclonal anti-CD72 primary antibodies (clones J4-117, 3F3 and BU40), and MMAF-conjugated secondary antibody. The cell lines used were RL (DLBCL, CD72+), SCC-3 (DLBCL, CD72-), Jeko-1 and Mino (both MCL, CD72+). The cell growth inhibition after 96 hrs of treatment for for all control conditions; anti-CD72 primary antibody, IgG isotype, MMAF secondary antibody alone or combined with IgG isotype; tested in all cell lines, CD72+ or CD72- ranged from -3 to +4%. Conversely, anti-CD72 primary antibodies plus MMAF secondary antibody had marked growth inhibition effect ranged from 24 to 80% on the three CD72+ cell lines and no effect on cell growth in CD72- cell line, suggesting internalization of CD72 and growth inhibition dependent on the presence of CD72 on the cell surface.

This is the first study demonstrating a potential role for CD72 as a novel target for DLBCL and MCL. These data suggest that CD72 may be a candidate for an ADC approach in DLBCL and MCL.

Cell line% growth inhibition anti-CD72 + MMAF 2°Anti-CD72 clone J4-117Anti-CD72 clone 3F3Anti-CD72 clone BU40SCC-3 (CD72-)-31-5RL (CD72+)503330Jeko-1 (CD72+)433024Mino (CD72+)778066

Citation Format: Erika Von Euw, Eileen Taschereau, Da Ming Ou, Monica Mead, Sarah Larson, Dennis Slamon. CD72 as a potential new therapeutic target in diffuse large B-cell lymphoma (DLBCL) and mantle cell lymphoma (MCL) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3041.
Abstract 3048: Targeting protein deglycosylation as a newly identified vulnerable point in melanoma
Authors: Lin Victor J.T. , Zolekar Ashwini , Mishra Nigam M. , Ho Yin Ying , Hayatshahi Hamed S. , Parab Abhishek , Sampat Rohit , Liao Xiaoyan , Hoffmann Peter , Liu Jin , Emmitte Kyle A. , Wang Yu-Chieh
Published: August 2019

As an enzyme that removes N-glycans from glycopeptides, N-glycanase 1 (NGLY1) deglycosylates denatured glycoproteins and allows proteasome-mediated protein degradation to efficiently occur. Although NGLY1 is known for this pivotal function, information regarding the responses of human cancer and normal cells to NGLY1 suppression is limited. Here, we examined how NGLY1 expression affects viability, tumor growth, and responses to therapeutic agents in melanoma cells and an animal model. Molecular mechanisms contributing to NGLY1 suppression-induced anticancer responses were revealed by systems biology and chemical biology studies. Using computational and medicinal chemistry-assisted approaches, we established novel NGLY1-inhibitory small molecules. Compared with normal cells, NGLY1 was upregulated in melanoma cell lines and patient tumors. NGLY1 knockdown caused melanoma cell death and tumor growth retardation. Targeting NGLY1 induced pleiotropic responses, predominantly stress signaling-associated apoptosis and cytokine surges, which synergize with the anti-melanoma activity of chemotherapy and targeted therapy agents. Pharmacological and molecular biology tools that inactivate NGLY1 elicited highly similar responses in melanoma cells. Unlike normal cells, melanoma cells presented distinct responses and high vulnerability to NGLY1 suppression. Our work demonstrated the significance of NGLY1 in melanoma cells, provided mechanistic insights into how NGLY1 inactivation leads to eradication of melanoma with limited impact on normal cells, and suggested that targeting NGLY1 represents a novel anti-melanoma strategy with the opportunity for a broad therapeutic window.

Citation Format: Victor J.T. Lin, Ashwini Zolekar, Nigam M. Mishra, Yin Ying Ho, Hamed S. Hayatshahi, Abhishek Parab, Rohit Sampat, Xiaoyan Liao, Peter Hoffmann, Jin Liu, Kyle A. Emmitte, Yu-Chieh Wang. Targeting protein deglycosylation as a newly identified vulnerable point in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3048.
Abstract 3090: In vivo characterization of AMG 510 - a potent and selective KRASG12Ccovalent small molecule inhibitor in preclinical KRASG12Ccancer models
Authors: Rex Karen , Saiki Anne Y. , Sun Ji-Rong , Holt Tyler , Koppada Neelima , Lanman Brian A. , Volak Laurie P. , Foti Robert S. , Cee Victor J. , Lipford J. Russell , Canon Jude
Published: August 2019

Somatic activating mutations of RAS family members are tumor driver mutations found in an estimated 21% of all cancers. Oncogenic KRAS mutations at residues G12, G13, and Q61 represent the most common RAS mutations found in solid malignancies. The prevalence of KRAS p.G12C tumors is ~13% of lung adenocarcinoma (including NSCLC), 3% of colorectal carcinoma (CRC), and 1% to 2% of numerous other solid tumors, representing an unmet medical need. We have developed AMG 510, an orally bioavailable, covalent inhibitor of KRASG12C with potent biochemical and cellular activity, and robust in vivo efficacy. AMG 510 inhibited SOS-catalyzed nucleotide exchange of recombinant mutant KRASG12C/C118A but had minimal effect on KRASC118A, which is wildtype at position 12. In cellular assays, AMG 510 covalently modified KRASG12C and inhibited KRASG12C signaling as measured by phosphorylation of ERK1/2 (p-ERK) in all KRAS p.G12C-mutant cell lines tested but did not inhibit p-ERK in cell lines with various other KRAS mutations. AMG 510 also selectively impaired viability of KRAS p.G12C mutant cell lines but did not affect cell lines with other KRAS mutations. In vivo pharmacodynamic assays demonstrated dose- and time-dependent inhibition of KRASG12Csignaling in human pancreatic and NSCLC tumor xenografts. Covalent modification of KRASG12C by AMG 510 was measured by mass spectrometry and correlated with p-ERK inhibition in tumors. AMG 510 significantly inhibited the growth of KRAS p.G12C xenografts and resulted in tumor regression. Combination treatment of AMG 510 with standard-of-care and targeted agents demonstrated enhanced tumor growth inhibition compared to either single agent. In a syngeneic model of KRAS p.G12C mutant cancer, AMG 510 treatment significantly inhibited tumor growth and caused regression. AMG 510 is currently in Phase 1, first-in-human clinical trials for patients with advanced solid tumors harboring a KRAS p.G12C mutation.

Citation Format: Karen Rex, Anne Y. Saiki, Ji-Rong Sun, Tyler Holt, Neelima Koppada, Brian A. Lanman, Laurie P. Volak, Robert S. Foti, Victor J. Cee, J. Russell Lipford, Jude Canon. In vivo characterization of AMG 510 - a potent and selective KRASG12Ccovalent small molecule inhibitor in preclinical KRASG12Ccancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3090.
Abstract 3121: Osteopontin-c is overexpressed and mediates cell adhesion and proliferation in leukemia cell line with KMT2A-AFF1
Authors: Bastos Ana Clara Santos da Fonseca , Santoro Júlio César , Blunck Caroline Barbieri , Ferreira Luciana Bueno , Pombo-de-Oliveira Maria do Soccro , Emerenciano Mariana , Gimba Etel R.
Published: August 2019

Osteopontin (OPN) primary transcript suffers alternative splicing generating at least five OPN splicing isoforms (OPN-SI) named OPNa, OPNb, OPNc, OPN4 and OPN5. In solid tumors, total OPN (tOPN) expression has been correlated to several cancer features. However, OPN-SI roles in hematological malignancies are still under investigation. A hallmark of B-cell acute lymphoblastic leukemia (B-ALL) is the recurrence of specific gene rearrangements associated with prognostic risk stratification. Patients harbouring the KMT2A-AFF1 fusion gene are included into poor prognostic subgroup, with an overall survival rate of 50% and increased risk of extramedullary sites involvement. In B-ALL, it has been reported that tOPN support tumor dormancy and cell survival in response to chemotherapeutic drugs. However, the specific roles of OPN-SI have not been addressed. The aim of this study was to analyze the expression of OPN-SI in B-ALL samples and then investigate the role of the most overexpressed OPN-SI on modulating cellular and molecular aspects of B-ALL presenting KMT2A-AFF1. We found that OPNc is overexpressed in relation to OPNa and OPNb in patients with KMT2A-AFF1. In addition, OPNc transcript levels were associated with poor prognostic features, such as central nervous system infiltration, white blood cell counting and the high-risk group at diagnosis. In response to OPNc silencing in B-ALL cell lines, we found increased proliferation rates, as opposed to decreased cell adhesion properties over matrigel matrix. Our data evidenced that overexpressed OPNc in B-ALL with KMT2A-AFF1 is correlated to poor prognostic features and may modulate the leukemic phenotype by the decrease of cell proliferation while promoting cell adhesion. Further work should be conducted in order to evaluate OPNc putative application as an additional risk-stratification and prognostic marker for B-ALL and how OPNc could control leukemia development and progression.

Note: This abstract was not presented at the meeting.

Citation Format: Ana Clara Santos da Fonseca Bastos, Júlio César Santoro, Caroline Barbieri Blunck, Luciana Bueno Ferreira, Maria do Soccro Pombo-de-Oliveira, Mariana Emerenciano, Etel R. Gimba. Osteopontin-c is overexpressed and mediates cell adhesion and proliferation in leukemia cell line with KMT2A-AFF1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3121.
Abstract 313: AXL confers intrinsic resistance to osimertinib and the emergence of tolerant cells
Authors: Taniguchi Hirokazu , Yamada Tadaaki , Wang Rong , Tanimura Keiko , Nishiyama Akihiro , Tanimoto Azusa , Adachi Yuta , Takeuchi Shinji , Yamaguchi Hiroyuki , Fukuda Minoru , Takayama Koichi , Mukae Hiroshi , Yano Seiji
Published: August 2019

A novel EGFR-tyrosine kinase inhibitor (TKI), osimertinib, has marked efficacy in patients with EGFR-mutated lung cancer. However, some patients show intrinsic resistance and an insufficient response to osimertinib. This study showed that osimertinib adversely stimulated AXL by inhibiting a negative feedback loop. Activated AXL was associated with EGFR and HER3 in maintaining cell survival and inducing the emergence of cells tolerant to osimertinib. AXL inhibition reduced the viability of EGFR-mutated lung cancer cells overexpressing AXL that were exposed to osimertinib. The addition of an AXL inhibitor during either the initial or tolerant phases reduced tumor size and delayed tumor re-growth compared to osimertinib alone. AXL was highly expressed in clinical specimens of EGFR-mutated lung cancers and its high expression was associated with a low response rate to EGFR-TKI. These results indicated pivotal roles for AXL and its inhibition in the intrinsic resistance to osimertinib and the emergence of osimertinib-tolerant cells.

Citation Format: Hirokazu Taniguchi, Tadaaki Yamada, Rong Wang, Keiko Tanimura, Akihiro Nishiyama, Azusa Tanimoto, Yuta Adachi, Shinji Takeuchi, Hiroyuki Yamaguchi, Minoru Fukuda, Koichi Takayama, Hiroshi Mukae, Seiji Yano. AXL confers intrinsic resistance to osimertinib and the emergence of tolerant cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 313.
Abstract 3136: Discovery of low abundance colorectal cancer related biomarkers by the ADAPT Biotargeting System
Authors: Hornung Tassilo , Zarkovic Jelena , Kassner Michelle , Rosenow Matthew , Miglarese Mark R. , Mayer Günter , Famulok Michael , Spetzler David B.
Published: August 2019

Disease biomarkers play an essential role in disease-diagnostics and in monitoring their responses to therapies. Identification of low to medium abundance biomarkers is one of the biggest challenges in the field. We have developed Adaptive Dynamic Artificial Poly-ligand Targeting (ADAPT), a method in which libraries of single-stranded oligodeoxynucleotides (ssODNs) are enriched for sequences that bind to targets existing in samples in various amounts. Pull-downs with enriched libraries followed by LC-MS/MS allows for the enrichment and identification of low abundance biomarkers that cannot be identified by conventional proteomics.

A highly diverse library of 1011 ssODNs was subjected to multiple rounds of positive selection against formalin-fixed paraffin-embedded (FFPE) colorectal cancer tissue with negative selection against adjacent non-cancer tissue of the same specimen. An enriched library of ~3x106 ssODNs that bound preferentially to cancer tissue was obtained and used in combination with LC-MS/MS to identify biomarkers related to colorectal cancer. Using this approach, a total of 14 proteins were identified in cancer but not in non-cancer tissue lysates. Their identification was only possible due to their enrichment by binding of the immobilized ssODN library to the cancer sample since the same proteins were undetectable by conventional proteomics. Immunohistochemical staining (IHC) of multiple colorectal cancer cases verified that several of the identified target proteins, including nucleophosmin (NPM1) and synaptotagmin-like protein 2 (SYTL2), showed higher expression in cancer tissue compared to adjacent non-cancer tissue.

Our data indicate the potential of the ADAPT platform to profile small differences between cancer affected and unaffected tissue and provide a novel approach for cancer biomarker discovery.

Citation Format: Tassilo Hornung, Jelena Zarkovic, Michelle Kassner, Matthew Rosenow, Mark R. Miglarese, Günter Mayer, Michael Famulok, David B. Spetzler. Discovery of low abundance colorectal cancer related biomarkers by the ADAPT Biotargeting System [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3136.