bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4805: Carcinogenesis by stem cell misplacement–a novel cancer theory and its implications
Authors: Wang Rui-An
Published: May 2015

For rather a long period of time, it has been widely accepted that cancer was caused by gene mutations which is termed somatic mutation theory (SMT). But now the SMT theory has met with increasing doubts, disbelieves, and been facing challenges from other alternative theories. I here from a pathological perspective, and with the evidences from molecular pathology, hisopathology, and epidemiology, show that the 80 year old paradigm of carcinogenesis process from atypia to in situ carcinoma to invasive cancer is wrong. Since 90% of the human cancers are epithelially derived “carcinomas”, the turnover of this paradigm naturally negates the SMT theory for SMT sits on this model. The invasive cancer grows out in the stroma de novo from misplaced stem cells rather than deriving from the transformed in situ carcinoma cells. Also, I will show evidences that cancer cells are not so-believed “apoptosis-resistant”, but rather “short-lived sick cells”. In fact, it is the increased cell death which makes the cancer cells proliferate ceaselessly to compensate for the cell loss, and move to new locations for better chance of survival. I design this novel cancer theory as SCMT, stem cell misplacement theory. This theory is backed by huge amount of solid evidences and will bring revolutionary conceptual changes to our understanding and treatment of cancer, e.g., in situ carcinomas are not genuine cancer biologically, thus should not be treated the same as cancer.

Citation Format: Rui-An Wang. Carcinogenesis by stem cell misplacement–a novel cancer theory and its implications. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4805. doi:10.1158/1538-7445.AM2014-4805
Abstract 4806: Angiogenesis is independently influenced by interstitial flow and concentration gradients of tumor secreted morphogens
Authors: Shirure Venktesh S. , George Steven C.
Published: May 2015

Angiogenesis facilitates growth and entry of tumors into the systemic circulation leading to metastasis. Morphogens, such as vascular endothelial growth factor (VEGF), secreted by the tumors are believed to guide nearby vasculature towards the tumor. The distribution of morphogens in tissues is affected by interstitial fluid flow, which also exerts forces in the tissue microenvironment. Numerous cells in the tumor microenvironment can sense and respond to force, yet our understanding of the independent effect of interstitial flow in tumor angiogenesis is limited. This is due, in part, to inadequate experimental models that cannot decouple spatial morphogen concentrations and interstitial flow. Notably, control of these parameters is not possible in animal models, and 2D cell systems lack the complexity of in vivo tissue. We addressed these limitations by designing a microfluidic platform of polydimethlsiloxane (PDMS) polymer consisting of three parallel microporous tissue compartments (< 0.1 mm3 total volume) that are media-fed with microfluidic lines. Endothelial colony forming cell-derived endothelial cells (ECFC-EC) and stromal fibroblasts in extracellular matrix proteins (e.g., fibrin and collagen) seeded in the central compartment formed interconnected microvascular networks with hollow lumens in 7-14 days. A mathematical model of mass and momentum transport in the microfluidic device was used to determine experimental conditions that either minimize the concentration gradients or the interstitial flow across the tissue chamber. The conditions were validated experimentally using various molecular size fluorescent dextrans. In the absence of a concentration gradient, microvessels demonstrated a striking bias for angiogenesis against the direction of the interstitial flow. Furthermore, the extent of migration against the flow increased with increasing interstitial flow. In separate experiments with minimal/no interstitial flow, angiogenisis is biased towards a positive concentration gradient of the morphogens such as VEGF. In conclusion, we have developed a novel microfluidic platform that can decouple the effects of concentration gradients and interstitial flow. Using this platform we show that angiogenesis is independently influenced by interstitial flow and concentration gradients of tumor secreted morphogens.

Citation Format: Venktesh S. Shirure, Steven C. George. Angiogenesis is independently influenced by interstitial flow and concentration gradients of tumor secreted morphogens. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4806. doi:10.1158/1538-7445.AM2014-4806
Abstract 4808: Aflibercept is highly active in clinically relevant, patient derived xenografts of colorectal cancer
Authors: Bagley Rebecca G. , Amalou Zakia , Chiron Marielle , Vuaroqueaux Vincent , Caron Anne , Baltes Nina , Fiebig Heinz H.
Published: May 2015

Aflibercept (ziv-aflibercept in USA), a new fusion protein, targets vascular endothelial growth factor (VEGF)-A, VEGF-B, and placental growth factor (PlGF). Aflibercept + FOLFIRI is approved for treating metastatic colorectal cancer (mCRC) that is resistant to or has progressed following an oxaliplatin containing regimen. We investigated the pharmacological activity of aflibercept in CRC patient derived xenografts (PDX), most metastases-derived, and characterized PDX molecular and phenotypic properties. In 48 CRC PDX, Whole Exome Sequencing revealed: 85% APC, 79% TP53, 51% KRAS, 3% NRAS, 25% PIK3CA, 11% PTEN and 8% BRAF mutants. There was no significant association between angiogenic factors, tumor mutation profiles, and response to aflibercept.

In tumors, protein levels of human (h) VEGF-A were significantly higher than mouse (m) VEGF-A, hPlGF, and mPlGF (Chiron et al, ESMO, 2013). Affymetrix probes detected VEGF-A and PIGF but not VEGF-B mRNA in most PDX. There was no significant association between expression of angiogenic factors and response to aflibercept. Histological sections of CRC tumors from PDX, xenografts created from cell lines, and patients’ tumor specimens indicated that stromal involvement in human tumors was recapitulated in PDX but not in tumors from cell lines. PlGF expression was analyzed by immunohistochemistry on colon cancer tissue arrays. A significant number of colon cancer specimens were positive for PlGF, expressed in tumor cells and stroma. Aflibercept resulted in anti-tumor activity in 47/48 CRC PDX models and complete tumor stasis in 31/48 models. Efficacy data from 5 PDX models responsive to aflibercept and CRC standard of care (SOC) drugs, all administered as monotherapies, are below. The robust activity of aflibercept in CRC PDX models that are sensitive to SOC drugs suggest that CRC patients would benefit from aflibercept treatment, independently of mutational status or expression of angiogenic factors.

In Vivo Sensitivity Profile (optimal Tumor/Control (%))CRC PDX ModelPrior TreatmentsAflibercept 25 mg/kg, SC, 2x/wk5-FU100 mg/kg, IP, 1x/wkIrinotecan 50 mg/kg, IV, 1x/wkOxaliplatin 10 mg/kg, IP, Q2WCetuximab30 mg/kg, IV,1x/wkKRAS StatusCFX 1991Not available-2.531.51.856.342.1MutantCFX 2032Not available3.147.76.760.535.7MutantCFX 2048Not available-10.326.221.350.12.6Wild-typeCFX 20655-FU, leucovorin-11.920.614.471.82.2Wild-typeCFX 2067None6.253.045.158.914.2Wild-type

Citation Format: Rebecca G. Bagley, Zakia Amalou, Marielle Chiron, Vincent Vuaroqueaux, Anne Caron, Nina Baltes, Heinz H. Fiebig. Aflibercept is highly active in clinically relevant, patient derived xenografts of colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4808. doi:10.1158/1538-7445.AM2014-4808
Abstract 4849: Implications of cancer induced blood coagulation in cancer diagnosis and therapy
Authors: Hisada Yohei , Yasunaga Masahiro , Hanaoka Shingo , Saijou Shinji , Sugino Takashi , Tsuji Atsushi , Saga Tsuneo , Tsumoto Kouhei , Manabe Shino , Kuroda Jun-ichiro , Kuratsu Jun-ichi , Matsumura Yasuhiro
Published: May 2015

A large body of clinical evidence indicates that abnormal coagulation occurs in a variety of cancers, especially invasive cancers. If cancer clusters erode adjacent normal or tumor vessels, hemorrhage may occur, and fibrin clots immediately form in situ to stop the bleeding. The fibrin clots are subsequently replaced by collagen, similar to the case in the normal wound healing process. Although there are many points in common between cancer-induced stroma and normal wound healing, the critical difference between the two is that the pathophysiological condition in cancer lasts for as long as cancer cells survive in the body (Adv Drug Deliv Rev 2012).

First, we successfully developed a mAb that reacts only with human insoluble fibrin, not human fibrinogen (Cancer Sci 2011). In contrast to previously reported anti-fibrin monoclonal antibodies (mAbs), our anti-fibrin clot mAb (clone 102-10) recognizes an unexplored hole that is uncovered only when a fibrin clot forms. The epitope of the 102-10 mAb was mapped to a hydrophobic region on the Bβ-chain that interacted closely with a counterpart region on the γ-chain in the soluble state. New anti-Bβ and anti-γ mAbs specific for peptides lining the discovered hole appeared to bind exclusively to fibrin clots. Analysis of the kinetics of fibrin clot deposition using 102-10 in several disease models indicated that fibrin clot formation occurs only in the acute phase of the disease process, thrombosis, inflammation or trauma, and that these clots virtually disappear within a few weeks to be substituted by collagen. Then, we developed a new PET probe using 102-10. The radiolabelled mAb was injected into mice bearing chemically induced spontaneous tumors, and the probe showed clear and specific accumulation in the tumors. We named this detection strategy cancer stromal targeting (CAST) diagnois. Immunohistochemistry using this mAb revealed higher fibrin deposition in WHO grade 4 gliomas than in lower-grade gliomas. Because erosive tumors are highly likely to show micro-hemorrhages, even early asymptomatic tumors detected with radiolabeled 102-10 mAb may be aggressively malignant (Sci. Rep. 2013).

In conclusion, CAST diagnosis and therapy based on anti-insoluble fibrin mAb may be a useful new strategy in the field of oncology.

Citation Format: Yohei Hisada, Masahiro Yasunaga, Shingo Hanaoka, Shinji Saijou, Takashi Sugino, Atsushi Tsuji, Tsuneo Saga, Kouhei Tsumoto, Shino Manabe, Jun-ichiro Kuroda, Jun-ichi Kuratsu, Yasuhiro Matsumura. Implications of cancer induced blood coagulation in cancer diagnosis and therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4849. doi:10.1158/1538-7445.AM2014-4849
Abstract 4863: Quantification of cell fusion events between breast cancer cells and breast epithelial cells
Authors: Mohr Marieke , Zänker Kurt , Dittmar Thomas , Edenhofer Frank
Published: May 2015

Recent data suggest that cell-cell fusion contributes to the development of hybrid cell clones showing genetic and behavioural alterations. But less is known about forces triggering cell fusion processes between breast cancer cells and tumor surrounding cells. The influence of chronic inflammation and tumor microenvironment is increasingly being recognized as an etiology for cancer progression and cell fusion. However, information on how and which components particularly affect cell-cell fusion and thereby disease outcome remains elusive.

Available facilities to quantitate these cell-cell fusion processes in vitro could provide a novel method to identify cancer promoting components of the stroma. In the present study cell-cell fusion was measured by using a cre/ loxP reporter system. Therefore, two breast cancer cell lines stable transfected with a double fluorescence reporter vector and a breast epithelial cell line containing cre recombinase were generated and cell fusion was promoted by co-culturing cells under appropriate conditions. Regarding the complex nature of tumor milieu the potential influence of several inflammatory and anti-inflammatory cytokines were tested and revealed alterations in fusogenic behaviour of cell lines. Increased fusion activity was observed while adding the proinflammatory cytokine TNFα or epithelial growth factor (EGF). It could also be assumed that hypoxia is a stimulus for cell-cell fusion. In contrast the chemoattractant SDF-1α which is involved in many cancer metastatic mechanisms hadn't any influence. Interestingly, both breast cancer cell lines showed markedly different fusion activity after activation of the transcription factor NfκB.

The present study establishes an efficient tool for quantification of cell-cell fusion processes in vitro by cre mediated recombination of a loxP reporter system. This highlights a new opportunity to identify a potential link between components priming and triggering cell-cell fusion and therefore carcinogenesis.

Citation Format: Marieke Mohr, Kurt Zänker, Thomas Dittmar, Frank Edenhofer. Quantification of cell fusion events between breast cancer cells and breast epithelial cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4863. doi:10.1158/1538-7445.AM2014-4863
Abstract 4870: Proteomics of phyllodes tumor revealed that decorin increase in the extracellular matrix by periostin deficiency decreased cancer cell motility and invasion
Authors: Ishiba Toshiyuki , Nakanishi Akira , Sato Takanobu , Nakagawa Tsuyoshi , Nagahara Makoto , Oono Ryo , Uetake Hiroyuki , Sugihara Kenichi , Miki Yoshio
Published: May 2015

The ability of cancer cells to metastasize is dependent on the interactions of their cell surface molecules with microenvironment. However the tumor microenvironment, especially cancer-associated stroma, is poorly understood. To search for proteins which are present in the stroma, we investigated the phyllodes tumor, which contains breast stromal tissue, specific expression proteins compared with normal tissue by using iTRAQ-based quantitative proteomic approach. Periostin and versican core protein were up-regulated in phyllodes tumor. Decorin, mimecan, hemoglobin subunit alpha, hemoglobin subunit beta and ketatin, type1 cytoskeletal 19 were up-regulated in normal tissue. Periostin and decorin are one of component of the extracellular matrix. Periostin upregulation has been reported in many cancer types and is consequently defined as a tumor-enhancing factor. On the other hand, decorin upregulation inhibits tumor growth by antagonizing tumor angiogenesis. Periostin upregulation in phyllodes tumor and decorin upregulation in normal tissue was validated by immunohistochemical analysis from phyllodes tumor thirty five patients. We determined the interaction between decorin and periostin in phyllodes tumor tissues and breast cancer cell line BT-20 cells, by using immunoprecipitation and mass spectrometry analysis. Furthermore, we uncovered the plasma membrane translocation of decorin from cytoplasm in BT-20 or T-47D cells by siRNA-mediated knockdown of periostin, and detected the secreted decorin to the extracellular medium using Multiple Reaction Monitoring (MRM) method. The periostin siRNA-treated BT-20 cells motility and invasion were prevented by the secreted decorin into the extracellular medium. The overexpression of decorin suppressed the metastatic breast cancer cell line, MDA-MB-231 cells motality and invasion. These results demonstrated the secreted decorin by periostin deficiency decreased cancer cells motality and invasion. This molecular mechanism may become the new target for anti-cancer therapy.

Citation Format: Toshiyuki Ishiba, Akira Nakanishi, Takanobu Sato, Tsuyoshi Nakagawa, Makoto Nagahara, Ryo Oono, Hiroyuki Uetake, Kenichi Sugihara, Yoshio Miki. Proteomics of phyllodes tumor revealed that decorin increase in the extracellular matrix by periostin deficiency decreased cancer cell motility and invasion. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4870. doi:10.1158/1538-7445.AM2014-4870
Abstract 4882: Expression profiles of proinflammatory chemokines in prostate cancer cell supernatants
Authors: Reyes Niradiz , Correa Oscar , Bettin Alfonso , Tiwari Raj K. , Geliebter Jan
Published: May 2015

Background: Prostate cancer progression is a complex process in which chemokines and their receptors play an important role related to tumor survival, growth, and metastasis.

Objectives: 1) To determine the expression profiles of proinflammatory chemokines in prostate cancer cell line supernatants and their association with the metastatic phenotype, and 2) To determine whether these chemokines are present in the exosomes released from prostate cancer cell lines examined.

Methods: Prostate cancer lines investigated included PWR-1E (non-tumorigenic), LNCaP (low metastatic potential) and PC-3 (high metastatic potential). The Human Common Chemokines Multi-Analyte ELISArray Kit (Qiagen) was used to evaluate the expression profiles of several proinflammatory chemokines in the supernatant and in the exosomal fraction of prostate cancer cells in culture. Chemokine profiles were correlated with the metastatic abilities of the cell lines analyzed. Chemokines evaluated were CCL2, CCL3, CCL4, CCL5, CCL11, CCL17, CCL22, CXCL1, CXCL8, CXCL9, CXCL10 and CXCL11.

Results: Differential chemokine profiles in the cell lines were observed in the supernatant and in the exosomal fractions.

Conclusions: Prostate cancer cell lines with different phenotypes have different chemokine profiles, both in supernatant and in the exosomal fraction of these cells in culture. These results may be useful for the design of diagnostic, prognostic and/ or therapeutic strategies.

Financial support: Universidad de Cartagena Project Code: Res 04670 - 2012

Citation Format: Niradiz Reyes, Oscar Correa, Alfonso Bettin, Raj K. Tiwari, Jan Geliebter. Expression profiles of proinflammatory chemokines in prostate cancer cell supernatants. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4882. doi:10.1158/1538-7445.AM2014-4882
Abstract 4889: Oncolytic adenovirus coated with stimuli responsive block copolymer mPEG-b-poly(His) induces tumor microenvironment-specific antitumor efficacy
Authors: Yun Chae-Ok
Published: May 2015

Adenovirus (Ad) has shown great promise in cancer gene therapy. Ad coated with cationic polymers have already been shown to enhance transduction, but it showed nonspecific binding with cells, resulted in evoked side effect. To overcome these hurdles, in this study adenovirus coated with pH sensitive poly(His) polymer through ionic interaction for passive tumor targeting. We have characterized the effect of polymer concentration on particle size distribution, zeta potential, transduction efficiency, and structural properties. Remarkable enhanced transduction efficiency was observed in cells treated with Ad/poly(His) nanocomplex compared to naked Ad at low pH. In addition, cellular trafficking using time lapse total internal reflection fluorescence showed that the Ad/poly(His) nanocomplex was significantly enhanced cellular uptake at lower pH, in contrast to neutral pH. The biodistribution study confirmed that the Ad/poly(His) nanocomplex can effectively accumulate at the tumor site by the EPR effect and exhibited 3.3-fold higher luciferase expressions in tumor than the naked Ad. Additionally, MTT assay and cancer cell xenografts studies showed that polymer coated nanocomplex exhibited high cancer cell killing effect and also inhibited the tumor growth. Furthermore, poly(His)-coated Ad complex was not significantly inhibited by serum, in contrast to naked Ad. Moreover, Ad/poly(His) nanocomplex significantly reduced the innate immune response in contrast to naked Ad, as assessed by interleukin-6 cytokine release from macrophage cells in vivo. These results suggest that the stimuli-responsive Ad/poly(His) nanocomplex has potential to targeting tumors.

Note: This abstract was not presented at the meeting.

Citation Format: Chae-Ok Yun. Oncolytic adenovirus coated with stimuli responsive block copolymer mPEG-b-poly(His) induces tumor microenvironment-specific antitumor efficacy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4889. doi:10.1158/1538-7445.AM2014-4889
Abstract 4893: USP7 modulates UV-induced PCNA monoubiquitylation by regulating DNA polymerase eta stability
Authors: Qian Jiang , Zhu Qianzhen , Pentz Kyle , He Jinshan , Wang Qien , Wani Altaf A.
Published: May 2015

DNA polymerase eta (Polη), a product of the Xeroderma pigmentosum variant (XPV) gene, plays a unique role in translesion DNA synthesis (TLS). It is a short-lived proteasomally degraded protein and its steady-state level is tightly controlled by multiple pathways. In this study, we have identified the deubiquitylating enzyme ubiquitin-specific protease 7 (USP7) as a novel regulator of Polη stability. Polη and USP7 interact in vitro and in vivo. Overexpression of wild-type USP7, but not its interaction-defective and catalytic mutants, deubiquitylate Polη and increase its steady-state level. The data demonstrate that both physical interaction and catalytic activity of USP7 are necessary for Polη deubiquitylation and stability regulation. Interestingly, knockout of USP7 also increased the steady-state levels and slowed the turnover of both Polη and p53, which is likely caused by destabilizing MDM2, a targeting E3 ligase for ubiquitylation of both Polη and p53. Furthermore, ectopic expression of USP7 upregulated the UV-induced PCNA monoubiquitylation. In contrast, UV-induced PCNA monoubiquitylation was not observed in XPV cells ectopically expressing USP7. Taken together, our results demonstrated that USP7 facilitates UV-induced PCNA monoubiquitylation by directly and indirectly regulating Polη stability. (The work was supported by grants from NIH.)

Note: This abstract was not presented at the meeting.

Citation Format: Jiang Qian, Qianzhen Zhu, Kyle Pentz, Jinshan He, Qien Wang, Altaf A. Wani. USP7 modulates UV-induced PCNA monoubiquitylation by regulating DNA polymerase eta stability. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4893. doi:10.1158/1538-7445.AM2014-4893
Abstract 4899: Small proline rich protein 3 (SPRR3) is a potential mediator of radiation resistance in HPV negative head and neck squamous cell carcinoma
Authors: Martin Joshua A. , Gunderson Dana , Kimple Randall J.
Published: May 2015

Purpose: Head and neck cancers (HNCs) are caused by two major etiologies that have significant prognostic significance: tobacco and human papillomavirus (HPV). Patients with HPV-positive HNC have significantly better outcomes than those with HPV-negative HNC. We have previously shown that this difference may be due, in part, to increased sensitivity to radiation in HPV-positive HNCs. In order to identify additional factors that may underlie differences in radiation sensitivity, we sought to understand radiation-induced changes in global gene expression between HPV-positive and HPV-negative HNCs.

Methods: A panel of eight cell lines, four HPV positive (UD-SCC2, UM-SCC47, UPCI-SCC90, 93-VU-147T) and four HPV negative (SCC1, SCC6, SCC22B, SCC1483 were treated with radiation (4 Gy) or mock and total RNA isolated 4, 24, and 48 hrs after treatment. Global gene expression was analyzed using Affymetrix Gene 1.1 ST array chips. Comparisons between untreated, baseline control, and post-radiation expression were made using ArrayStar. Alterations in differentially expressed genes were validated by Real-Time quantitative PCR (RT-qPCR).

Results: Out of the top 15 hits, several members of the small proline rich repeat (SPRR) family were identified. SPRR1A, SPRR1B, and SPRR3 were significantly upregulated in HPV negative as compared to HPV-positive cell lines. Differences in expression were validated using RT-qPCR in a panel of HNC cell lines.

Conclusions: We identified changes in expression of SPRR3 in HPV negative cell lines. This gene is a member of a protein family known to scavenge reactive oxygen species. Its increased expression in HPV negative HNC may mediate resistance to radiation and contribute to the poor outcomes of patients in this cohort. SPRR3 is a potential novel molecular target that could improve the outcomes of patients with HPV negative HNC.

Citation Format: Joshua A. Martin, Dana Gunderson, Randall J. Kimple. Small proline rich protein 3 (SPRR3) is a potential mediator of radiation resistance in HPV negative head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4899. doi:10.1158/1538-7445.AM2014-4899
Abstract 4900: The role of Interleukin (IL)-22 on the UVB-induced proliferation and inflammation of human keratinocyte cell line, HaCaT
Authors: Kim Yejin , Kim Hyemin , Choi Jiwon , Jang Mirim , Choi Jiyea , Kang Jae Seung , Lee Wang Jae
Published: May 2015

Interleukin 22 (IL-22), a member of IL-10 family, is a potent mediator of inflammatory responses. It is produced by activated CD4+ T cells and natural killer (NK) cells, but IL-22Rα expression is restricted to nonhematopoietic cells in the skin, pancreas, intestine, liver, lung and kidney. IL-22 suppresses IL-4 production from Th2 cells and induces acute phase proteins in liver and pancreas. It is recently found that IL-22 plays a critical role in the maintenance of epidermal homeostasis by controlling cell cycle of keratinocytes. Therefore, we investigated that role of IL-22 on the proliferation and inflammation of ultraviolet B (UVB)-irradiated human keratinocytes, HaCaT. First, we found the expression of IL-22Rα mRNA and protein level was increased in HaCaT by UVB (100 J/m2) irradiation. Interestingly, the translocation of IL-22Rα from cytosol to membrane was increased by UVB irradiation. When UVB-irradiated HaCaT was cultured in the presence of recombinant IL-22 (rIL-22), the suppressed proliferation of HaCaT by UVB was rescued. In addition, we found that the production of UVB-induced pro-inflammatory cytokines, such as IL-1α, IL-6 and IL-18, was increased by the treatment of rIL-22. Taken together, IL-22 plays a critical role in inflammation by UVB irradiation. And it suggests that UVB-induced skin inflammation could effectively be controlled by the regulation of IL-22 production and its action on keratinocytes.

Note: This abstract was not presented at the meeting.

Citation Format: Yejin Kim, Hyemin Kim, Jiwon Choi, Mirim Jang, Jiyea Choi, Jae Seung Kang, Wang Jae Lee. The role of Interleukin (IL)-22 on the UVB-induced proliferation and inflammation of human keratinocyte cell line, HaCaT. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4900. doi:10.1158/1538-7445.AM2014-4900
Abstract 4901: Clostridium cluster XIVa species an early biomarker in radiation injury
Authors: Ahmed Ishfaq , McLauglin William , Anant Shrikant , Umar Shahid , Papineni Rao V.
Published: May 2015

Radiation therapy is being used in the treatment of nearly 60% of the current cancer patients. In spite of the best efforts to provide special care in minimizing radiation exposure, significant radiation injury remains a common side-effect particularly to the gastro intestinal tract. Severe injury to GI tract may lead to multiple organ dysfunction syndrome. The status of fecal microbiota has been proposed as a rapid diagnostic platform in radiation injury. Gut microbes play a role in immunity, health, and disease. Imbalances in gut microbiota are related to numerous disorders, such as inflammatory bowel disease, colorectal cancer (CRC), and atopy. In patients with immune suppression due to high-dose chemotherapy or that undergoing radiation therapy, disruption of the microbiota through antibiotics and impairment of host immunity gives rise to perturbations favoring intestinal domination by pathogenic species, resulting in increased bacterial translocation and susceptibility to systemic infection. Here, using real-time PCR assays, we assessed the immediate changes and quantitative alterations to fecal microbiota in response to radiation injury. Swiss-albino Mice were subjected to 10 Gy X-ray whole body irradiation doses using commercially available X-RAD 320 irradiator (1Gy/min; F2 beam hardening filter 1.5mm Al, 0.25mm Cu, 0.75mm Sn; Precision X-ray inc, USA). Fecal samples collected immediately after the radiation exposure were analyzed. We show that Clostridium cluster XIVa that accounts for almost 60% of the mucin-adhered microbiota is significantly increased (11.71 +/- 0.29 fold n=6) in the fecal samples, compared to the total bacteria and other bacteria that tend to enrich in the luminal region. Along with the Bacteroidetes and Proteobacteria from the luminal region, minimal changes in the levels of Firmicutes that colonize the mucin layer were observed. Further, these changes varied considerably between the subjects and were inconsistent compared to the Clostridium cluster species. Clostridium cluster XIVa has been proposed to be the mucosal butyrate producers producing butyrate close to the epithelium. This enhances butyrate bioavailability required in treating diseases such as inflammatory bowel disease. These findings provide opportunities to evaluate the potential use of intestinal microbiota as biomarkers in radiation injury. Also, provides insights into the microbial changes tantamount to the radiation induced GI injury.

Citation Format: Ishfaq Ahmed, William McLauglin, Shrikant Anant, Shahid Umar, Rao V. Papineni. Clostridium cluster XIVa species an early biomarker in radiation injury. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4901. doi:10.1158/1538-7445.AM2014-4901
Abstract 4906: mTOR kinase inhibition enhances radiosensitivity in glioblastoma
Authors: Tandle Anita T. , Tofilon Philip , Camphausen Kevin
Published: May 2015

Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor and predicts a poor prognosis despite current treatment modalities. Aberrant activation of the phosphatidyl-inositol-3-kinase (PI3K) pathway is observed in nearly 90% of GBMs making the downstream effector, mammalian target of rapamycin (mTOR), a potential drug target. mTOR is a serine/threonine kinase that regulates cell growth, proliferation, survival, protein translation and metabolism by integrating signals received from the PI3 K/Akt signaling pathway. The allosteric mTOR inhibitor rapamycin has failed in the clinic as a treatment for GBM patients. Therefore, in the current study, we examined the potential benefit of MLN0128, a novel potent mTOR ATP competitive inhibitor, as a therapeutic strategy for GBM. We assessed in vitro antiproliferative activity of MLN0128 in a panel of GBM tumor and stem cell lines. MLN0128 significantly inhibited cell proliferation of both GBM tumor and stem cells. It decreased the phosphorylation of mTOR, Akt, S6, 4E-BP1, and NDRG1, inhibiting both mTORC1 and mTORC2 signaling in GBM. Thus, MLN0128 appears to be superior in blocking mTORC1/2 signaling in contrast to rapamycin. It inhibited migration and invasion of GBM cells in a matrigel coated chamber assay. On clonogenic survival assay MLN0128 decreased GBM cell survival and increased radiosensitivity. To investigate the mechanism of radiosensitization, the induction and repair of DNA double-strand breaks were evaluated by γH2AX foci formation. MLN0128 exposure significantly delayed the dispersal of radiation induced γH2AX foci formation in both tumor and stem cells. These results indicate that MLN0128 enhances tumor cell radiosensitivity by inhibiting DNA repair. Next we combined MLN0128 with temozolomide (TMZ), a FDA approved radiosensitizer for GBM therapy. Although, we did not observe any synergy in inhibition, addition of TMZ did not adversely affect MLN0128 inhibitory effects. These findings support the rationale for clinical testing of MLN0128 in patients with GBM to provide insight towards optimizing therapeutic efficacy of mTOR kinase inhibitors with standard care.

Note: This abstract was not presented at the meeting.

Citation Format: Anita T. Tandle, Philip Tofilon, Kevin Camphausen. mTOR kinase inhibition enhances radiosensitivity in glioblastoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4906. doi:10.1158/1538-7445.AM2014-4906
Abstract 4912: Hyperthermia induced NO repressed NFκB regulates IR-induced EMT, CSC self-renewal and pluripotency
Authors: Dinges Corinne , Natarajan Mohan , Aravindan Sheeja , Herman Terence S. , Aravindan Natarajan
Published: May 2015

Based on clinical evidence, hyperthermia treatment (HT) in combination with radiation (IR) achieves superior local control for recurrent chest wall breast cancer (BCa). The effect of HT combined with IR is due partly to its independent cytotoxic effects, and partly to its radiosensitizing effect. Recently, we have shown that apoptosis upon hyperthermia of BCa cells is regulated by NO-mediated suppression of NFκB. Also, we have demonstrated that tumor cells that survive a course of IR are instrumental in tumor relapse and progression. Accordingly, we investigated whether HT exerts NO-NFκB axis dependent regulation of IR-induced alterations in factors that drive the EMT, CSCs self-renewal capacity and pluripotency maintenance in surviving BCa Cells. Human breast adenocarcinoma (MCF-7) cells either mock-irradiated or exposed IR (2Gy) with or without HT (43°C for 1h) were analyzed for transcriptional regulation of 93 stem cell related molecules using QPCR profiling. NFκB (p50/p65) overexpression, muting by ΔIκBα (with IR) and NO induction by SNP (with HT) approach were used to delineate the role of HT-induced NO and the selective targeting of IR-induced NFκB. Nanog, Sox-2, ABCG-2, N-Cadherin and Myc expressions were examined with immunoblotting. IR profoundly increased the transactivation of 80 stem-cell related molecules. Interestingly, HT reverted the activation of 79 IR-induced genes. Attenuation of all 80 IR-induced genes observed after blocking NFκB with ΔIκBα and by increasing intracellular NO with SNP demonstrated the significant role of NO dependent NFκB suppression in HT-induced inhibition of IR modulated stem-cell related transcriptome. Expression levels of Nanog, Sox-2, ABCG-2, N-Cadherin and Myc validates the potential of HT in mitigating IR-induced stemness. Together this data demonstrates that HT attenuates IR-induced stemness in BCa cells and further imply that HT induced increase in NO may alleviate stemness by selectively targeting IR-induced NFκB.

Citation Format: Corinne Dinges, Mohan Natarajan, Sheeja Aravindan, Terence S. Herman, Natarajan Aravindan. Hyperthermia induced NO repressed NFκB regulates IR-induced EMT, CSC self-renewal and pluripotency. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4912. doi:10.1158/1538-7445.AM2014-4912
Abstract 4924: Mucolytic drug N-Acetylcystein improves 89Zr-trastuzumab HER2-immunoPET imaging by restoring accessibility to HER2 hampered by membrane mucin overexpression in a HER2 positive MUC4 overexpressing xenograft mouse model
Authors: Wimana Zena , Gebhart Geraldine , Morandini Renato , Flamen Patrick , Ghanem Ghanem
Published: May 2015

89Zr-trastuzumab, a 89-Zirconium radiolabeled anti-HER2 monoclonal antibody, has been shown appropriate to enable positron emission tomography (PET) imaging of HER2 positive lesions in breast cancer (BC) patients. Binding of 89Zr-trastuzumab to the HER2 receptor, a prerequisite could be undermined by one of the postulated resistance mechanisms to trastuzumab, i.e. the interference of cell membrane glycoproteins mucins. This study aimed to assess whether mucolytic drug N-Acetylcystein (NAC) could enhance the uptake of 89Zr-trastuzumab in a HER2 positive and MUC4 overexpressing xenograft mouse model.

Materials & Methods: Trastuzumab was first coupled with the chelate N-succinyl-desferal ester, subsequently N-SucDF-trastuzumab was radiolabelled with 89Zr-oxalate in a HEPES buffered solution and purified by size exclusion chromatography. The impact of NAC on 89Zr-trastuzumab PET was studied in a dual model xenograft generated by s.c. inoculation of HER2+/MUC4- SKBr3 and JIMT1 HER2+/MUC4- BC cell lines, with the latter overexpressing MUC4. Mice were treated with NAC p.o. during 6 days. Body distribution and uptake of the tracer were measured 1) by in vivo PET/CT imaging and by 2)ex vivo tissue analysis.

Results: In mice treated with NAC, there was a significant increase of the Standardized Uptake Value (SUV) mean, SUVmax and SUV Volume Histogram (SVH) in the MUC4 overexpressing tumors compared to the control group. This was not the case for MUC4 negative SKBR3 tumors. Furthermore, ex vivo body distribution data supported these results with a significant increase of radioactivity uptake in the MUC4 positive tumors and no change in non-tumoral tissues

Conclusion: N-Acetylcysteine may be an interesting drug to improve the accessibility of trastuzumab to its receptors, HER2, by affecting mucin masking.

Note: This abstract was not presented at the meeting.

Citation Format: Zena Wimana, Geraldine Gebhart, Renato Morandini, Patrick Flamen, Ghanem Ghanem. Mucolytic drug N-Acetylcystein improves 89Zr-trastuzumab HER2-immunoPET imaging by restoring accessibility to HER2 hampered by membrane mucin overexpression in a HER2 positive MUC4 overexpressing xenograft mouse model. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4924. doi:10.1158/1538-7445.AM2014-4924
Abstract 493: NRF2 modulates sensitivity to thymidylate synthase inhibitors in colon cancer cells
Authors: Clinton Sarah A. , Barbour Karen W. , Ozer Ufuk , Berger Franklin G.
Published: May 2015

Thymidylate synthase (TYMS) catalyzes the reductive methylation of dUMP, and is the sole de novo source of thymidine for DNA replication and repair. As such, it is an important target of chemotherapeutic drugs, particularly the fluoropyrimidines 5-fluorouracil (FUra) and 5-fluoro-2’-deoxyuridine (FdUrd), as well as the folate analog raltitrexed (RTX). In cells, these drugs are metabolized to derivatives that bind to and inhibit TYMS, leading to depletion of thymidine levels, dysregulation of redox metabolism, generation of oxidative stress, and, eventually, apoptotic cell death. Gene expression profiles of FUra-treated HCT116 cells (a human colon cancer cell line) showed that a number of genes involved in redox metabolism are up-regulated following drug exposure. Several of these were recognized as known targets of nuclear factor-erythroid-2-related factor 2 (NRF2), a member of the cap ‘n’ collar family of bZIP transcription factors that confers protection against oxidative and electrophilic stress. Quantitative PCR assays verified that FUra, FdUrd, and RTX do indeed induce several NRF2-regulated genes, including AKR1B10, ALDH3A1, NQO1, and SERPINE1, among others. Overproduction of NRF2 via DNA transfection increased expression of some, but not all, of these genes, a phenomenon that was reversed by Kelch-like ECH-associated protein-1 (KEAP1), a known negative regulator of NRF2. The impact of TYMS inhibitors on expression of NRF2 target genes was maintained in the presence of exogenous thymidine and the antioxidant N-acetylcysteine, even though these agents completely ameliorated drug-induced apoptosis. This suggests that the response of NRF2 target genes to TYMS-directed drugs occurs independently of TYMS inhibition and subsequent oxidative stress. Utilizing siRNA technology, we examined the impact of NRF2 knockdown on drug-induced apoptosis, and found that increases in the apoptotic index following exposure to TYMS inhibitors were greater in NRF2-knockdown cells as compared to control cells. This was also observed in TYMS-overproducing cells that are profoundly drug resistant, and indicates that reduced NRF2 expression sensitizes cells to these TYMS inhibitors. Overall, we conclude that the induction of NRF2 target genes is a constraining factor in the cytotoxic response to TYMS inhibitors, and that inhibition of the transcription factor may be an effective strategy to enhance sensitivity to these agents.

Citation Format: Sarah A. Clinton, Karen W. Barbour, Ufuk Ozer, Franklin G. Berger. NRF2 modulates sensitivity to thymidylate synthase inhibitors in colon cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 493. doi:10.1158/1538-7445.AM2014-493
Abstract 4933: In vivo visualization of heterogeneous cancer cell populations by fluorescence molecular tomography
Authors: Zanca Ciro , Inda Maria del Mar , Bonavia Rudy , Wykosky Jill , Verkhusha Vladislav , Cavenee Webster , Furnari Frank
Published: May 2015

Glioblastoma is the most common and aggressive tumor type that affects the central nervous system in adults. Despite advances in treatment strategies, its prognosis remains poor. EGFR and its mutant form EGFRvIII are amplified in a subset of clinical cases and interaction between cell populations expressing each receptor has been shown to enhance overall tumor growth. For instance, EGFRvIII induces expression of several secreted oncogenic factors, such as IL6 and LIF, which in turn stimulate the growth of EGFR expressing cells through an IL6 receptor (gp130)/EGFR cross-talk interaction. Therapy directed towards EGFR or EGFRvIII inhibition has been ineffective because of mechanisms that inhibit cell death. We hypothesize that these factors secreted by EGFRvIII expressing cells, in addition to promoting tumor growth, play a role in the failure of EGFR-directed therapies reported for glioblastoma. Preliminary data indicate that EGFRvIII-expressing cells prompt an IL6/LIF-mediated paracrine and autocrine antiapoptotic signaling cascade, leading to upregulation of the pro-survival protein, Survivin. RNAi-mediated downregulation of Survivin reverts this phenotype. To determine the effect of identified signaling effectors and cytokines in promoting EGFR tyrosine kinase inhibitor resistance, an in vivo model system of heterogeneous tumor growth was established and imaged using fluorescence molecular tomography (FMT) technology. Survivin, upstream pathways and cytokines were ablated in EGFR or EGFRvIII-expressing glioma cell lines tagged with different near infrared fluorescent proteins (turboFP635 or iRFP713). Orthotopically engrafted tumors of mixed EGFR/EGFRvIII populations were then evaluated for their tumor growth and cellular composition when challenged with EGFR tyrosine kinase inhibitors. In conclusion, this system allows for the validation of molecular mechanisms that promote cancer cell survival and establishes the basis for alternative treatment strategies.

Citation Format: Ciro Zanca, Maria del Mar Inda, Rudy Bonavia, Jill Wykosky, Vladislav Verkhusha, Webster Cavenee, Frank Furnari. In vivo visualization of heterogeneous cancer cell populations by fluorescence molecular tomography. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4933. doi:10.1158/1538-7445.AM2014-4933
Abstract 4942: Metastatic tumor migration in lymphatic vessels revealed by real-time intravital imaging
Authors: Fink Darci M. , Connor Alicia L. , Kelley Philip M. , Tempero Richard M. , Hollingsworth Michael A.
Published: May 2015

Distant tumor metastases and their confounding symptoms are often responsible for reduced patient quality-of-life and decreased overall survival. The main route of metastasis for many solid tumors is the lymphatic vasculature. The mechanisms of metastatic spread of tumor cells through the lymphatic vessels remain poorly understood. We have developed a novel transgenic mouse model and customized real-time intravital microscopy platforms to study the migration of tumor cells into lymphatic vessels in live animals over the course of several days. TdTomato fluorescent protein was expressed in lymphatic endothelial cells using an inductive CRE recombinase approach. This transgenic system coupled with the use of fluorescent stereomicroscopy techniques allowed us to visualize the cutaneous lymphatic vessel network with remarkable anatomic detail in living mice. We tracked the fate of CFSE-labeled B16 tumor cells administered cutaneously using two-color intravital fluorescence microscopy. This strategy allowed us to simultaneously visualize the lymphatic vessel network (red) and the B16 tumor cells (green) at single-cell resolution. We clearly visualized and documented several key behaviors of metastasizing tumor cells including single-cell intravasation, movement of tumor cells within vessels, clearing from vessels, and the accumulation of tumor cells in regional lymph nodes. These events occurred at a low frequency over the course of days in conditions of tissue homeostasis. Current studies are designed to define the anatomic site of tumor intravasation into lymphatic vessels and investigate how the inflamed tumor microenvironment affects tumor migration into the lymphatic vasculature.

Citation Format: Darci M. Fink, Alicia L. Connor, Philip M. Kelley, Richard M. Tempero, Michael A. Hollingsworth. Metastatic tumor migration in lymphatic vessels revealed by real-time intravital imaging. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4942. doi:10.1158/1538-7445.AM2014-4942