bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract LB-158: A human xenograft model for developing personalized targeted therapy for pancreatic cancer
Authors: Walters Dustin M. , Adair Sara J. , Stokes Jayme B. , Borgman Cheryl , Lowrey Bryce , Adams Reid B. , Fox Jay W. , Papin Jason A. , Parsons J. Thomas , Bauer Todd W.
Published: February 2011

Background: Effective treatment regimens for pancreatic ductal adenocarcinoma (PDAC) are currently lacking. Molecular-targeted therapies, while promising, have been ineffective likely due to the heterogeneity and redundancy of molecular signaling pathways. Identification of predictors of response to defined targeted therapies is critical for the development of effective and personalized therapies.

Methods: We have established a human PDAC tumor bank and have orthotopically implanted patient tumors into the pancreas of immunocompromised mice. Clinicopathologic data has been collected for each patient's tumor. Tumors successfully propagated in mice are routinely screened for six genetic mutations commonly found in pancreatic cancer. Gene expression profiling and phospho-receptor tyrosine kinase (pRTK) arrays, cytokine arrays, and reverse phase protein arrays (RPPAs) are used to profile the activated signaling pathways in individual tumors. A unique genetic and molecular profile is then constructed for each human tumor. Ingenuity pathway analysis software (Ingenuity Systems, Inc., Redwood, CA) is being implemented to map the unique signaling network for each tumor using the genetic and molecular data.

Results: To date 7 tumors have been analyzed, including 2 metastatic tumors and 5 primary adenocarcinomas. All tumors contained TP53 gene mutation, whereas 5 tumors exhibited mutated KRAS and 3 contained mutations in the SMAD4 gene. Analysis using pRTK arrays revealed universal activation of EGFR and variable activation of ErbB2/Her2, RON, Tie-2, FGF-R1 and 3, and Axl. The profile of pRTKs appeared stable upon tumor passage both in vivo and in culture. RPPAs revealed significant variation among tumors in activation for MAPK, AKT, SAPK, and p38. Additional cytokine/growth factor array analysis and gene expression profiling is currently underway.

Conclusion: The establishment of an orthotopic human pancreatic cancer xenograft model allows for the systematic analysis of genetic and molecular characteristics of human tumors propagated within a relevant tumor microenvironment. This model affords the ability to assess tumor growth responses to targeted therapies currently in use for multiple malignancies and to correlate tumor response with genetic perturbations, changes in gene expression and activation of cell signaling networks. This model will be a significant step toward developing personalized targeted therapy for pancreatic cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-158.
Abstract LB-162: The onset of MYCN overexpression-induced neuroblastoma is accelerated by ALK mutation in zebrafish
Authors: Lee Jeong-Soo , Zhu Shizhen , Shin Jimann , Perez-Atayde Antonio R. , Feng Hui , Guo Feng , Helman Daniel , Wang Wenchao , Stewart Rodney A. , George Rani , Kanki John , Look A. Thomas
Published: February 2011

Neuroblastoma (NB) is the most common extracranial solid tumor in infancy and originates in the peripheral sympathetic nervous system (PSNS). High-risk NB is fatal in the majority of patients, despite intensive myeloablative chemotherapy. The MYCN oncogene is amplified in over 20% of NB, particularly in those with highest risk treatment failure. We have developed a zebrafish NB model by overexpressing human MYCN under the control of the dopamine-beta-hydroxylase (D) promoter specific for noradrenergic cells in the PSNS. Fish from this stable transgenic line developed tumors as early as 4 months of age with approximately 20% penetrance at 8 months of age. The tumors resemble human NB histologically, immunohistochemically, and ultrastructurally. The expression of MYCN suppressed the normal development of PSNS neurons and chromaffin cells in the head kidney during embryogenesis and in young adult fish. In some fish, tumor cells began to repopulate the interrenal gland of the head kidney by 2 months of age. Germline and somatic activating mutations have been identified in the ALK gene, which encodes a receptor tyrosine kinase, in human NB, including those with MYCN amplification. To assess cooperativity between amplified MYCN and mutant ALK genes in transformation, we generated a zebrafish stable transgenic line in which the activated mutant form of ALK (F1174L) was expressed under the control of the DßH promoter. These transgenic animals did not display an abnormal phenotype nor did they develop tumors during the first 6 months of life. In contrast, mutant ALK expression accelerated the onset of MYCN-induced neuroblastoma when the 2 genes were co-expressed in double transgenic fish, indicating that MYCN over-expression and activating ALK mutations can cooperate in tumorigenesis. Although co-expression of mutant ALK accelerated the onset of tumors, it did not rescue the MYCN-induced suppression of PSNS development, suggesting that an as-yet-unidentified tumor suppressor gene, possibly located on distal 1p in the human genome, may be lost to fulfill this role in NB tumorigenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-162.
Abstract LB-168: Genistein sensitizes radio-resistant cancer cells through regulation of NF-kappaB
Authors: Kim Su-Hyeon , Song Yong-Sang
Published: February 2011

Background: NF-kappaB is a key transcription factor involved in the control of normal cellular and organismal processes, such as immune and inflammatory responses, developmental processes, cellular growth, and apoptosis. However, NF-kappaB is persistently active in a number of disease states, including cancer. It has been reported that chemo- or radio- resistant cancer cells had elevated NF-kappaB activity. Recent studies including ours have showed that genistein, a soy-derived isoflavonoid, has antitumor activity in various cancer cells.

Objective: In this study, we evaluated whether genistein could sensitize ionizing radiation resistance of cervical cancer cells through regulation of NF-kappaB.

Methods: Six cervical cancer cell lines (HeLa, CaSki, C33A, ME180, SiHa, HeLa229) were treated by 20 Gy radiation with 50 M genistein or not. Cell growth rate was measured by MTT assay and protein expression level was analyzed by western blotting. Protein localization was observed by confocal microscopic analysis. NF-kappaB transcriptional activity was measured by luciferase reporter gene assay. NF-kappaB DNA binding activity was evaluated by electophoretic mobility shift assay.

Results: Among six cervical cancer cells, HeLa and HeLa229 cells showed radio-resistance accompanied by elevated NF-kappaB activity. Ionizing radiation induced NF-kappaB nuclear transclocation and stimulated NF-kappaB DNA binding and transcriptional activities. However, pretreatment with genistein inhibited radiation induced transcriptional activity coincident to decreased radio-resistancy. Genistein also inhibited radiation-induced NF-kappaB regulated anti-apoptotic gene expression including c-myc, Survivin, XIAP, Bcl-2, IAP-1, TRAF, A1/Bfl-1 and FLIP.

Conclusion: Taken together, genistein can sensitize radio-resistant cancer cells through regulation of NF-kappaB and could be used as a radio-sensitizer for the treatment of cervical cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-168.
Abstract LB-172: Transgene presence in patients’ tumors following tumor-targeted nanodelivery
Authors: Pirollo Kathleen F. , Nemunaitis John J. , Senzer Neil N. , Sleer Leanne , Chang Esther H.
Published: February 2011

Discussant: William F. Benedict, UT M. D. Anderson Center, Houston, TX.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-172.
Abstract LB-18: The Cullin 3 substrate adaptor DIP-2 mediates DAPK ubiquitination to control interferon responses
Authors: Shen Che-Hung , Lee Yu-Ru , Ho Hsuan-Chung , Yuan Wei-Chien , Chen Chun-Hau , Shih Hsiu-Ming , Chen Ruey-Hwa
Published: February 2011

Death-associated protein kinase (DAPK) is a pro-apoptotic, calmodulin-regulated serine/threonine kinase and was originally identified based on its involvement in interferon- -induced cell death. Whether and how interferon-γ-signaling regulates the activity or expression of DAPK remains largely unexplored. In this study, we identify the BTB-Kelch protein DIP2 functions as a negative regulator of DAPK by potentiating the proteosomal degradation of DAPK. DIP2 binds DAPK and Cul3 via its Kelch repeats and BTB domain, respectively. The DIP2-Cul3-ROC1 E3 ligase complex promotes DAPK ubiquitination both in vivo and in vitro, and downregulation of endogenous DIP2 diminishes DAPK ubiquitination and biological functions. The DIP2-dependent DAPK ubiquitination is suppressed in cells receiving either interferon-α or interferon-γ, which induces an enrichment/sequestration of DIP2 in the PML nuclear bodies, thereby separating DIP2 from its substrate DAPK. Through disrupting the interaction of DAPK with its E3 ligase, interferon triggers the stabilization of pro-apoptotic DAPK, which contributes to the cell death-promoting function of interferon. This study not only identifies the DIP2-Cul3-ROC1 complex as an E3 ligase for DAPK ubiquitination, but also reveals the underlying mechanism of DAPK regulation by interferon.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-18.
Abstract LB-182: Serum amyloid A is elevated in serum of lung cancer patients with poor prognosis
Authors: Cho William Chi Shing , Cheng Wai-wai , Yip Tai-tong , Au Joseph SK
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-182.
Abstract LB-184: Identification of breast cancer susceptibility genes using whole exome sequencing
Authors: Thompson Ella , Ellul Jason , Li Jason , Doyle Maria , Campbell Ian
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-184.
Abstract LB-195: Anti-CD40 and CpG-ODN enhance the antimelanoma response of hu14.18K322A in vivo.
Authors: Alderson Kory L. , Luangrath Mitchell , Gillies Stephen D. , Navid Fariba , Rakhmilevich Alexander L. , Sondel Paul M.
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-195.
Abstract LB-198: Collagen remodeling and tissue mechanics at different stages of mammary tumor development
Authors: Yu Hongmei , Weaver Valerie M.
Published: February 2011

Discussion

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-198.
Abstract LB-207: IFN-Γ directly regulates ifi202B expression to maintain CTL proliferation potential and tumor rejection efficacy
Authors: Zimmerman Mary A. , Liu Kebin
Published: February 2011

A common procedure in human cytotoxic T lymphocyte (CTL) adoptive transfer cancer immunotherapy is to propagate tumor-specific CTLs in vitro using anti-CD3 mAb prior to transfer. One of the major impediments of CTL adoptive cancer immunotherapy is lack of CTL persistence in the tumor-bearing host after adoptive transfer. Therefore, maintaining CTL proliferation and persistence in the tumor microenvironment is of great significance in cancer immunotherapy. In this study, we observed that stimulation of tumor-specific CTLs with anti-CD3 mAb results in loss of proliferative potential and poor tumor-rejection efficacy in vivo, whereas, stimulation with tumor-specific antigen maintains CTL proliferative potential and persistence, leading to potent tumor rejection in an experimental lung metastasis mouse model. Analysis of genome-wide gene expression profile revealed that ifi202B, a gene with known function in regulating T cell proliferation and apoptosis, was dramatically up-regulated in antigen-stimulated but not in anti-CD3 mAb-stimulated CTLs. Furthermore, although both antigen- and anti-CD3 mAb-stimulated CTLs express similar level of IFN-Γ receptor, antigen stimulation led to rapid activation of STAT1, a key mediator of the IFN-Γ signaling pathway, whereas anti-CD3 mAb stimulation failed to activate STAT1. We further demonstrated that activated STAT1 binds to the promoter of ifi202b gene in T cells. Taken together, these observations suggest that IFN-Γ produced by the activated CTLs and the IFN-Γ signaling pathway plays a critical role in maintaining CTL proliferation and persistence in the tumor microenvironment through regulating ifi202B.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-207.
Abstract LB-230: Automating sample preparation for single-cell protein phosphorylation analysis by flow cytometry
Authors: Liu Li , Eluwa Kelechi , Grom Patricia , Yoder Amy , Kowalski Michael , Song Yong , Shankey T. Vincent
Published: February 2011

Protein phosphorylation plays essential roles in regulating cellproliferation, differentiation and other cellular processes. Aberrant regulation of protein kinases, can drive malignancy through inappropriate cell growth and survival signals. Understanding the phosphorylation status of specific cellular proteins in cancer has proven valuable in developing targeted therapies and assays to identify specific drug responses in individual cancer patients.

The ability to analyze cells at the single cell level allows flow cytometry to measure protein phosphorylation in rare subsets of cells in complex populations such as blood or bone marrow specimens. However, sample preparation for protein phosphorylation analysis by flow cytometry can be technically challenging, time-consuming, and subject to error when performed manually. Furthermore, the procedure demands strict control of experimental conditions such as time and temperature due to the dynamic nature of protein phosphorylation. In this report, we describe an automated sample preparation method using a @Biomek® NXP Span-8 work-station to monitor the phosphorylation status of ERK and p38 MAPK in human peripheral blood monocytes. The method allows automation of sample preparation steps, including 1) ERK/p38 MAPK stimulation, 2) fixation, 3) erythrocyte lysis, 4) cell permeabilization, and, 5) staining with phospho-specific and cell surface marker antibodies. Using this approach, we were able to observe p38 MAPK and ERK activation in CD14+ monocytes following lipopolysaccharide (LPS) or phorbol-12-myristate-13 acetate (PMA) treatment. The automated sample preparation workstation is capable of processing 48 samples in approximately 2.5 hours.

Note: *Method cited Not for Clinical purposes. @For Laboratory Use Only; not for use in diagnostic procedures.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-230.
Abstract LB-245: Nuclear translocation of HO-1 in prostate cancer. Role beyond heme degradation
Authors: Gueron Geraldine , Ferrando Mercedes , Elguero Belen , Salles Angeles , Meiss Roberto , Colombo Lucas , Curino Alejandro , Facchinetti Maria Marta , Navone Nora , De Siervi Adriana , Vazquez Elba
Published: February 2011

Prostate cancer (PCa) is the second leading cause of cancer-associated death in men. The metastatic spread of PCa cells and the ability to survive when reaching the metastatic nitch is affected by growth factors, chemokines, angiogenic factors and hormones. Heme oxygenase-1 (HO-1), the inducible isoform of the rate-limiting enzyme in heme degradation, counteracts oxidative and inflammatory damage. Here we investigated its nuclear translocation in androgen sensitive (MDAPCa2b and LNCaP) and insensitive (PC3) PCa cell lines. Our results show that endogenous levels of HO-1 are markedly lower in PC3 compared to MDAPCa2b and LNCaP. Hemin treatment, a potent and specific inducer of HO-1, increased protein levels with a resulting nuclear translocation in PCa cell lines. Stable transfection of HO-1 in PC3 (PC3HO-1) also showed a significant nuclear translocation. Furthermore, human prostate cancer PC3HO-1 cells growing subcutaneously in athymic nude mice showed significant HO-1 nuclear staining (detected by immunohistochemsitry and immunofluorescence) compared to PC3pcDNA3 xenografts. Using RT-qPCR-generated gene array we observed a set of inflammatory and angiogenic genes up- or down-regulated in response to HO-1 overexpression, identifying VEGFA as a novel downstream target of HO-1. HO-1 modulation limited the metastatic potential of neoplastic cells by down-regulating VEGFA production. An in vivo angiogenic assay also demonstrated that PC3HO-1 tumors presented less neovascularization than tumors derived from control cells, giving further evidence to support HO-1 anti-angiogenic function. These results implicate for the first time HO-1 in the regulation of key proinflammatory factors, involved in PCa progression. Taken together our results suggest that HO-1 may be a potential target for therapeutic interventions.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-245.
Abstract LB-25: Regulation of Np63α by IκB kinase-β
Authors: Chatterjee Aditi , Chang Xiaofei , Sen Tanusree , Ravi Rajani , Bedi Atul , Sidransky David
Published: February 2011

The p53 family (p53, p73, p63) plays an instrumental role in the cellular stress response, including induction of cell cycle arrest and apoptosis in response to DNA damage. In addition to p63 and p73 isotypes capable of transactivating downstream target gene expression (TA isotypes), both genes also express dominant negative inhibitory isoforms, such as ΔNp63α. ΔNp63 is degraded in response to DNA damaging agents, thereby enabling an effective cellular response to genotoxic agents. Here, we identify a key molecular mechanism underlying the regulation of ΔNp63α expression in response to extrinsic stimuli, such as chemotherapeutic agents or TNF-α. We show that ΔNp63α interacts with IκB kinase (IKK), a multisubunit protein kinase that consists of two catalytic subunits, IKKα and IKKβ, and a regulatory subunit, IKKγ (NEMO-NF-κB essential modifier). We find that IKK kinase promotes ubiquitin-mediated proteasomal degradation of ΔNp63α, whereas a kinase-deficient mutant IKK -K44A fails to do so. Cytokine- or chemotherapy-induced stimulation of IKKβ leads to degradation of ΔNp63α and augments trans-activation of p53 family-induced genes involved in the cellular response to DNA damage. Conversely, inhibition of IKKβ with a NEMO-binding peptide or siRNA-mediated silencing IKKβ expression attenuates cytokine- or chemotherapy induced degradation of ΔNp63α. These data demonstrate that IKKβ plays an essential role in regulating ΔNp63α in response to extrinsic stimuli. Our findings suggest that the activation of IKK may be a mechanism by which levels of ΔNp63α are reduced, thereby rendering the cells susceptible to cell death in the face of cellular stress or DNA damage.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-25.
Abstract LB-260: Sensitivity of breast cancer stem cells to TRA-8 anti-DR5 monoclonal antibody
Authors: Joshi Angelina I. Londoño , Oliver Patsy G. , Forero Andres , LoBuglio Albert F. , Buchsbaum Donald J.
Published: February 2011

Breast cancer stem cells (BrCSC) are reported to be responsible for self-replication, treatment resistance, and metastases. Doubly enriched populations of BrCSC have been isolated using combined stem cell markers for CD44+/CD24- and increased levels of ALDEFLUOR® positivity based on aldehyde dehydrogenase enzyme activity. We have examined the sensitivity of doubly enriched populations of BrCSC isolated from basal-like breast cancer cell lines as regards anti-DR5 agonistic monoclonal antibody (TRA-8) mediated cytotoxicity. Methods: BrCSC were isolated and characterized from basal breast cancer cell lines (SUM159, SUM149, HCC38 and 2LMP) based on the intracellular accumulation of ALDEFLUOR® reagent and CD44+/CD24- surface expression by flow cytometry. DR5 expression on the BrCSC subpopulation, in vitro cytotoxicity after 24 h exposure to TRA-8, and caspase activation after TRA-8 exposure were evaluated. The effects of TRA-8 on secondary tumorosphere formation were also investigated. Results: Unsorted basal-like cell lines were highly sensitive to TRA-8 mediated cytotoxicity with IC50 values of 0.7-3.0 ng/ml. Doubly enriched CD44+/CD24−/ALDEFLUOR+ BrCSC from these cell lines exhibited comparable DR5 expression and were similarly highly sensitive to TRA-8 induced cytotoxicity with rapid induction of caspase 3 and 8 cleavage. Secondary tumorosphere formation was greater than 90% inhibited by TRA-8 exposure. An agonistic anti-DR4 monoclonal antibody (2E12) had no effect, and TRAIL was only partially inhibitory to secondary tumorosphere formation at equimolar concentrations. TRA-8 effect on BrCSC tumorigenicity will be presented. These studies suggest that TRA-8 may be an effective targeted therapy directed at BrCSC in basal-like breast cancer patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-260.
Abstract LB-27: Systematic identification of genes contained the translational regulation element IRES in the cancer development
Authors: Tseng Joseph Ta-Chien
Published: February 2011

Internal Ribosome Entry Site (IRES) is a cis-acting element located at the 5′ untranslated region to regulate the protein translation. Translation by internal ribosome entry was first identified in picornaviruses, but a number of cellular mRNAs containing IRESs have subsequently been found, including basic fibroblast growth factor, c-myc, p53, and HIF-1. Till now, around 80 cellular IRES elements have been identified, almost half found in genes encoding oncogenes, growth factors, and proteins involved in the regulation of apoptosis. This suggests that IRES-dependent translation is an important regulatory mechanism for the maintenance of cellular homeostasis, especially when cap-dependent initiation is inhibited. Therefore, it would be interested to understand how many genes bearing the IRES to elucidate the functional role of IRES. In this study, the pattern search algorithm was used to screen the whole human transcripts to identify the genes bearing the IRES elements. A special phenomenon which multiple IRES structures existed in specific region was found, and this region was named as internal ribosome entry zone (IREZone). The total human transcripts were identified to contain the IRES or IREZone element about 24.8% and 45.6%, respectively. By using bicistronic assay, the activity of IRES zone is higher then the single IRES existed in the 5′UTR of c-jun. Via Ingenuity software, we found that the cancer-related genes carry multiple IRES or IREZone in their 5′UTR. This result indicated these cancer-related genes are more tightly regulated at translational level to express the protein in the right time. And the mis-regulation of these proteins expression may lead to the cancer development.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-27.
Abstract LB-274: Clinical significance of MUC1, MUC2 and CK17 expression patterns in the diagnosis of pancreatic carcinoma
Authors: Yang Haisu , Tamayo Rosalba , Horten Bruce , DaSilva Moacyr , Gangi Maryann , Vazquez Evelyn , Joseph Daisy , Okamoto Patricia , Scholl Thomas
Published: February 2011

Pancreatic cancer is characterized by aggressive growth and treatment resistance. Identification of unique biomarkers for diagnosis and prognosis is important in the treatment of this disease. In this study, we investigated the expression patterns of MUC1, MUC2 and CK17 in normal tissues as well as metastatic adenocarcinomas by immunohistochemical analysis. Our results clearly show that MUC1 (pan-epithelial membrane mucin), MUC2 (intestinal-type secretory mucin) and CK17 can be utilized as a panel of markers to collectively distinguish pancreatobiliary carcinoma from other primary site carcinomas. That is, tumors originating in the pancreatobiliary system showed an expression pattern of MUC1 (+), MUC2 (-) and CK17 (+). In contrast, tumors that arose from the colorectal region were MUC1 (-), MUC2 (+) and CK17 (-), while those originating from the lung and kidney had a MUC1 (+), MUC2 (-) and CK17 (-) expression profile. More importantly, the MUC1 (+), MUC2 (-) and CK17 (+) result displayed greater sensitivity and specificity than CA19-9, which is the currently accepted and widely used pancreatic tumor marker for the diagnosis of pancreatic cancer. Taken together, our data support an antibody panel that can enhance the current methods for pancreatic cancer diagnosis by specifically identifying the primary tissue of origin from which pancreatic carcinomas are derived.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-274.
Abstract LB-318: Development of Listeria monocytogenes-based vaccine that concomitantly targets both the tumor cells as well as tumor vasculature
Authors: Wallecha Anu , Rivera Sandra
Published: February 2011

Recently we described the development of Listeria monocytogenes (Lm) based chimeric human Her2/neu gene (ChHer2) harboring most of the known MHC class I epitopes of the HER2/neu oncogene (ChHer2) when expressed as a fusion protein to a non-hemolytic fragment of listeriolysin O. This construct when tested for immunogenicity and anti-tumor effects in mice proved to be efficacious in breaking immune tolerance toward the HER2/neu self-antigen and elicited strong T cell immune responses in experimental animals. These results support the further clinical development of this vaccine for treatment of HER2/neu overexpressing malignancies such as breast, colorectal and pancreatic cancers. Preclinical studies with three different Lm based vaccines targeting angiogenesis antigens have demonstrated potential in mediating the regression of tumors arising from distinct origin such as breast or skin cancer. Therefore, we hypothesized that the delivery of an angiogenic antigen simultaneously with a tumor-associated antigen will likely have a synergistic effect by impacting on tumor growth via targeting blood vessels while at the same time targeting a tumor specific antigen to enhance effector T cell infiltration in the tumor and improve the vaccine therapeutic efficacy. If true, this will result in a flexible platform by using different tumor antigens. We have created two dual recombinants Lm vaccines that express and secrete both the tumor specific antigen such as ChHer2 and tumor vasculature antigen such as High molecular weight melanoma associated antigen (HMW-MAA) successfully. Currently we are evaluating the anti-tumor effects and immune responses generated by these constructs. If successful, this study will have a great potential for human use in cancer immunotherapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-318.
Abstract LB-324: A toll-like receptor 4 agonist enhances the efficacy of trastuzumab therapy and promotes adaptive immunity and long-term protection against a human ErbB-2 (HER2)-transfected syngeneic tumor in a human HER2 transgenic mouse model
Authors: Wang Shangzi , Xu Wei , Tang Yong , Surana Rishi , Bingham Catherine , Astsaturov Igor A. , McCarthy Kenneth , Alpaugh R. Katherine , Littlefield Bruce A. , Hawkins Lynn D. , Ishizaka Sally T. , Weiner Louis M.
Published: February 2011

Antibodies against cancer-relevant targets can recruit Fc receptor-bearing myeloid and natural killer cells to mediate antibody dependent cellular cytotoxicity (ADCC). Toll-like receptors are potent activators of the innate immune system and generate signals leading to the initiation of an adaptive immune response that can be utilized for therapeutic purposes. We tested the efficacy of E6020, a TLR4 agonist, administered in combination with the clinically approved anti-HER2 antibody trastuzumab. A four-week course of therapy with trastuzumab and E6020 protected against the outgrowth of syngeneic D5 melanoma cells transfected with the human tumor antigen HER2 in immunodeficient C57BL/6 SCID mice, in immunocompetent wild-type C57BL/6 mice as well as in human HER2-tolerant C57BL/6 hmHER2 transgenic mice. In each tested strain of mice, the addition of E6020 to trastuzumab significantly increased the proportion of animals that survived initial tumor challenge compared to trastuzumab treatment alone (83% vs. 33% in C57BL/6 hmHER2 transgenic mice; p < 0.05). To determine if apparently cured mice had been effectively immunized against human HER2, the transgenic mice were inoculated subcutaneously with 103 D5-HER2 cells and then received a 4-week course of trastuzumab+E6020, or trastuzumab alone. Animals that survived the primary tumor challenge were followed for at least 120 days and then rechallenged with 5 × 103 D5-HER2 or unmodified D5 cells and followed for a minimum of 60 additional days. Mice that had been initially treated with the combination of trastuzumab and E6020 demonstrated superior protection (22/30; 73%) to those treated with trastuzumab alone (16/36; 44%; p < 0.05) against rechallenge with D5-HER2, but no protection was seen against D5 challenge. Both CD4 and CD8 T-cells were required for full protection against rechallenge. These findings suggest that trastuzumab therapy can induce a host-protective adaptive immune response. Moreover, combined treatment with trastuzumab and a TLR4 agonist enhances both the direct anti-tumor effects of trastuzumab and a host-protective human HER2-directed adaptive immune response.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-324.
Abstract LB-330: Endogenous T cell responses to lung adenocarcinoma neoantigens delay malignant tumor progression
Authors: DuPage Michel , Cheung Ann F. , Mazumdar Claire , Winslow Monte M. , Bronson Roderick T. , Crowley Denise , Jacks Tyler
Published: February 2011

Neoantigens derived from somatic mutations in tumors may provide a critical link between the adaptive immune system and cancer. However, it has been difficult to properly recapitulate immune responses against tumor neoantigens in models of the human disease. Here we describe a novel system to induce the expression of potent neoantigens in a genetically engineered mouse model of lung cancer. We show that endogenous T cells respond to and infiltrate tumors, resulting in a significant delay in malignant progression. However, despite continued neoantigen expression, T cell infiltration does not persist and tumors ultimately escape immune attack. In contrast, transplantation of cell lines derived from lung tumors that express neoantigens, even those that have escaped the immune system in their autochthonous setting, or prophylactic vaccination against autochthonous tumors, evoke robust T cell responses that either lead to rapid tumor eradication or selection of tumors that lose neoantigen expression. These results support clinical data that suggest a role for the immune system in cancer suppression rather than prevention. This model provides a valuable system for the optimization of immune therapies that can eradicate human cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-330.
Abstract LB-334: IFN-γ and TRAIL cooperate to enhance colon carcinoma apoptosis and immune cell-mediated anti-tumor cytotoxicity
Authors: Liu Feiyan , Liu Kebin
Published: February 2011

Development of cancer-selective therapy with little or no toxicity is one of the goals of modern translation cancer research. TRAIL selectively induces apoptosis in various types of tumor cells but not in normal cells, and thereby is a promising selective anti-cancer agent. Currently, TRAIL is in early stage clinical trials for treatment of almost all types of human cancers. However, tumor cell resistance to TRAIL is a major impediment of TRAIL therapy. Most current approach to overcome TRAIL resistance is by pharmacologic means, which often involve use of toxic therapeutic agents. In this study, we demonstrated here that IFN-γ, a cytokine secreted by activated immune cells, modulate the mitochondrion apoptosis pathway to sensitize human colon carcinoma cells to TRAIL-induced apoptosis. IFN-γR is expressed in human colon carcinoma, and moderate to high positive correlations were found between cell surface IFN-γR level and TRAIL-induced apoptosis. Furthermore, activated CD8+ T cells express high levels of IFN-γ and TRAIL, and TRAIL is also expressed in tumor-infiltrating CD4+, CD8+ T cells, macrophage and NK cells. Consequently, blocking TRAIL function significantly increased colon carcinoma metastasis in vivo. Our data suggest that cooperation between IFN-γ and TRAIL play critical role in anti-tumor immunity.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-334.
Abstract LB-34: Targeted delivery of siRNAs to diverse solid cancers by a novel immunoconjugate comprising tetrameric human protamine linked to an internalizing anti-TROP-2 humanized antibody
Authors: Gupta Pankaj , Goldenberg David M. , Chang Chien-Hsing
Published: February 2011

Despite the potential of RNA interference (RNAi) to silence specific genes, the full therapeutic potential of RNAi remains to be realized due to the lack of an effective delivery system to target cells in vivo. To address this critical need and prompted by the promising results obtained with fusion proteins comprising the nucleic-acid-binding human protamine and a cell-targeting Fab or scFv, we have developed novel immunoconjugates having multiple copies of human protamine tethered to a tumor-targeting, internalizing antibody for targeted delivery of siRNAs in vivo. The Dock-and-Lock (DNL) method developed by us provides the design and strategy for building such multifunctional agents. DNL enables site-specific, self-assembly of two modular components with each other, resulting in a stable covalent structure of defined composition with retained bioactivity. The basic strategy of DNL involves the generation of two types of modules, one containing the dimerization and docking domain (DDD) of cAMP-dependent protein kinase (PKA) and the other containing the anchoring domain (AD) of a related A-kinase anchoring protein (AKAP). Docking a DDD-containing module with an AD-containing module occurs spontaneously, and the resulting complex is locked with disulfide bonds to enhance in vivo stability. We generated DDD2-L-thP1, a DDD-module of truncated human protamine (thP1, residues 8 to 29 of full human protamine 1), in which the sequences of DDD2 and thP1 are fused to the light chain of a humanized antibody at the N- and C-termini, respectively. The reaction of DDD2-L-thP1 under mild redox conditions with hRS7-IgG-AD2, an AD-module derived from an anti-TROP-2, internalizing, humanized antibody, resulted in the facile formation of the desirable DNL complex, designated E1-L-thP1, which contains four copies of thP1 appended to the carboxyl termini of the heavy chains. The purity and molecular integrity of E1-L-thP1 following Protein A purification was shown by size-exclusion HPLC and SDS-PAGE. In addition, the ability of E1-L-thP1 to bind plasmid DNA or siRNAs was demonstrated by the gel shift assay. E1-L-thP1 was effective in protecting the bound DNA from digestion by nucleases added to the sample or present in serum, and its function to carry siRNAs into TROP-2 expressing cancer cells was confirmed by fluorescence microscopy using FITC-conjugated siRNAs and the human lung cancer cell line, Calu-3. Further evaluation of the in vitro and in vivo efficacy of E1-L-thP1 for delivery of CD74-specific siRNAs to treat TROP-2 expressing pancreatic cancer is ongoing.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-34.
Abstract LB-354: Dysregulation of microRNA-34a expression in head and neck squamous cell carcinoma promotes tumor growth and tumor angiogenesis
Authors: Kumar Bhavna , Yadav Arti , Teknos Theodoros N. , Kumar Pawan
Published: February 2011

Objectives of the study: Head and Neck squamous cell carcinoma (HNSCC) results from deregulation of multiple signaling pathways leading to tumor cells acquiring proliferative, invasive, angiogenic and metastatic properties. Interactions between neoplastic cells and the surrounding microenvironment also play a major role in tumor progression. Recent studies have demonstrated that microRNA's (miRs) play a critical role in cancer development where they can act as oncogenes or as onco-suppressors. A microRNA array analysis of a panel often HNSCC cell lines reveled that miR-34a is significantly downregulated in all these cell lines as compared to normal human oral keratinocyes (HOK). In addition, miR-34a expression was also downregulated in highly angiogenic endothelial cells (endothelial cells overexpressing Bcl-2; EC-Bcl-2) as compared to normal human endothelial cells. However, little is known about the role of miR-34a in HNSCC tumor growth and tumor angiogenesis. In this study, we examined if dysregulation of miR-34a in HNSCC promotes tumor growth and tumor angiogenesis.

Methodology: The study was performed using a panel of established human head & neck cancer cell lines. miRs expression in tumor and endothelial cells was examined by miR-arrays and further validated by real time PCR. Lipofectamine-2000 was used to transfect miR-34a in 2 HNSCC cell lines (UM-SCC-74A and UM-SCC-47) and human endothelial cells. Cell-proliferation assay was performed by MTT and colony formation assay. Tumor cell and endothelial cell motility was assessed by scratch assay. Expression of miR-34a downstream mediators was measured by Western blot.

Results: Ectopic expression of miR34a significantly inhibited cell proliferation of UM-SCC-74A cells (52% and 58% on days 2 and 4) and UM-SCC-47 cells (43% and 47% on days 2 and 4). In addition, enhanced expression of miR34a in UM-SCC-74A and UM-SCC-47 cells significantly inhibited tumor cell colony formation and tumor cell migration. Similar to tumor cells, miR-34a significantly inhibited EC-Bcl-2 proliferation and migration by down regulating c-met, Bcl-2 expression and upregulating phospho p38 MAPK expression.

Conclusion: These findings indicate that dysregulation of miR-34a expression is common in HNSCC and modulation of miR34a activity might represent a novel therapeutic strategy for the treatment of head and neck cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-354.
Abstract LB-355: miR-29a suppresses tristetraprolin, which is a regulator of epithelial polarity and metastasis
Authors: Gebeshuber Christoph
Published: February 2011

Several microRNAs (miRNAs) have recently been described as crucial regulators of epithelial-to-mesenchymal transition (EMT) and metastasis. By comparing the expression profiles of miRNAs, we found up regulation of miR-29a in mesenchymal, metastatic RasXT cells relative to epithelial EpRas cells. Overexpression of miR-29a suppressed the expression of tristetraprolin (TTP), a protein involved in the degradation of messenger RNAs with AU-rich 3′-untranslated regions, and led to EMT and metastasis in cooperation with oncogenic Ras signalling. We also observed enhanced miR-29a and reduced TTP levels in breast cancer patient samples, indicating relevance for human disease. Previously, miR-29 family members were shown to have tumor-suppressive effects in haematopoietic, cholangiocytic and lung tumors. Therefore, miRNAs can act as either oncogenes or tumor suppressors, depending on the context.

In addition to miR-29a we found several other miRNAs in our miRNA profiling analyses of cell lines and murine tumors and could confirm them and regulated targets in human breast cancer patient samples.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-355.
Abstract LB-388: Macrophage migration inhibitory factor (MIF) plays a role in proliferation, differentiation, and survival of Ewing tumor cells through the activation of several kinases
Authors: Kang Hyung-Gyoo , Liu Xian Fang , Sorensen Poul HB , Triche Timothy J.
Published: February 2011

Cytokines and chemokines are involved in various mechanisms of cell signaling networks, which control cell growth, proliferation, differentiation, and survival. In normal cells, expression of cytokines and their downstream cell signaling pathways are tightly and accurately regulated. However, it has been shown that cytokine signaling pathways are deregulated in many tumors. Thus, identification and understanding of deregulated cytokines and their related pathways may provide unique and promising opportunities for better-targeted therapies of cancer. In the previous experiments we showed that Macrophage migration Inhibitory Factor (MIF) is highly expressed in Ewing tumor cells and it is involved in the proliferation and survival of Ewing tumors. Our present data implies that MIF might functionally contribute to Ewing tumor cell growth, differentiation and survival through the activation of several kinases such as AMPK (AMP activated protein kinase), Paxillin and PYK2 (Protein Tyrosine Kinase2). In conclusion, coupled with earlier and present studies about MIF-dependent effects on Ewing tumors our data suggested that MIF might be involved in various pathways of Ewing tumor biology and might be a promising target of Ewing tumor cells therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-388.
Abstract LB-391: The modified effect of obesity on the association between height and cancer incidence
Authors: Lee Sang-Ah , Kim Jong-Hoon , Sung Joohon
Published: February 2011

Height has been regarded as anthropometric marker of early-life environments and reported the association with the risk of cancer incidence and mortality although the results were not consistent for some site-specific cancers. This study was aimed to evaluate the association between height and cancer incidence among Korean and the modification effects at BMI on the association. In addition, the association between height and the mortality of 5 major cancer sites was analyzed.

The study participants of 1,067,923 Koreans (609,560 men and 458,363 women) aged 30-65 years was included in the follow-up or cancer incidence and mortality between 1993 and 2005. Study participants were divided into quartile according to their height. The lower quartile served as the reference group. Hazards ratio (HR) and 95% confidence intervals (CIs) associated with height included in the model were age, weight, cigarette smoking habits, alcohol drinking habits, physical activity, economic status, age at menarche, and menopause status.

After adjustment for all the covariates, each 5 cm increment in height was associated with 5% and 7% higher risk of overall cancer incidence in men and women, respectively. In case of site-specific cancers, a positive association was BMI (under vs. over 25 kg/m2), the effect of height on the risk of lung and prostate cancer incidence was predominantly observed among obese women only.

These results suggest the association between height and cancer incidence might be modified by BMI status.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-391.