bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4536: Combination external beam and internal radiation via 131I-CLR1404 in the treatment of head and neck squamous cell carcinoma xenografts.
Authors: Saker Jarob , Armstrong Eric , Bednarz Bryan , Farhoud Mohammed , Besemer Abigail , Weichert Jamey , Harari Paul
Published: August 2014

Radiotherapy (RT) is a central treatment modality for head and neck cancer (HNC). However, optimal outcome is commonly limited by the radiation tolerance of adjacent normal tissue structures. Radiation is sometimes valuable in the setting of locoregional tumor recurrence, however the risk of significant normal tissue toxicity is substantial with repeat irradiation and can have a profound adverse impact on patient quality of life. Although technical advances including intensity modulated radiotherapy (IMRT) can reduce normal tissue effects, the problem of repeat exposure of normal tissues to radiation is frequently limiting. The radiolabeled drug 124/131I-CLR1404 is a phospholipid ether analogue (PLE) that accumulates preferentially within malignant tumors and enables internal delivery of radiation. This allows a combination of external and internal radiotherapy delivery with a goal of improving treatment outcome without enhanced normal tissue toxicities.

In this study, we characterized the ability of 131I-CLR1404 to augment RT response using xenografted mice harboring human squamous cell carcinomas (SCC). We determined tumor growth delay as well as tumor and normal tissue dose distribution of 131I-CLR-1404 via Monte Carlo simulation using PET data generated with 124I-CLR1404 imaging. Tumor growth delay was extended by 15.5 days using combined 3.7MBq 131I-CLR1404 and RT compared to RT or 131I-CLR1404 alone. In combination therapy, single dose administration of 7.4 MBq 131I-CLR1404 as well as split dosing with 2 x 3.7 MBq separated by one week showed the same additional growth delay of 27 days compared to single dose of 3.7 MBq. Monitoring drug distribution via PET/CT showed high tumor selectivity with a tumor-to-liver ratio of 6.2±0.65 and a tumor-to-muscle ratio of 6.7±0.21.

These data illustrate the tumor selectivity of the CLR1404 PLE delivery platform as well as augmentation of tumor response with combined RT and 131I-CLR1404 treatment. De-intensification of external beam radiation dose, with attendant reduction in normal tissue toxicities so common in HNC patients represents a central objective to be tested in controlled clinical trials with the ultimate goal of improving treatment outcome and quality of life for HNC patients.

Citation Format: Jarob Saker, Eric Armstrong, Bryan Bednarz, Mohammed Farhoud, Abigail Besemer, Jamey Weichert, Paul Harari. Combination external beam and internal radiation via 131I-CLR1404 in the treatment of head and neck squamous cell carcinoma xenografts. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4536. doi:10.1158/1538-7445.AM2013-4536
Abstract 4555: Design and discovery of novel small molecule redox modulators as therapies for pancreatic cancer.
Authors: Pathania Divya , Sechi Mario , Palomba Michele , Sanna Vanna , Berrettini Francesco , Sias Angela , Taheri Laleh , Neamati Nouri
Published: August 2014

Pancreatic cancer is a complex disease. Late stage detection, poor prognosis, and resistance to available chemotherapeutics highlight the need to discover new and potent anticancer agents for its effective treatment. Altered cellular bioenergetics and oxidative stress are emerging hallmarks of most forms of cancer including pancreatic cancer. Cancer cells are more prone to reactive oxygen species (ROS)-mediated cell death due to their inherent elevated basal oxidative stress as compared to normal cells. Increased basal ROS levels could represent the Achilles’ heel of cancer cells. Even though cancer cells have adapted to survive in an oxidative stress environment, exogenous oxidative insults can overwhelm the adaptive antioxidant capacity of cancer cells and trigger ROS-mediated cell death

We have identified a novel class of compounds that act as redox modulators and specifically induce cell death in cancer cells. Compound 3a was identified through a medium throughput screen of ∼1000 highly diverse in-house compounds and chemotherapeutic agents for their ability to alter cellular bioenergetics using XF 24 extracellular flux analyzer (Seahorse Bioscience, Billerica, MA). Further structural optimizations led to the discovery of a more potent analog, 3b that displayed anti-proliferative activities at low micromolar levels in both drug-sensitive and drug-resistant cancer cell lines. Treatment with compound 3b causes Akt activation resulting in increased cellular oxygen consumption, oxidative stress and depletion of cellular antioxidant pool in pancreatic cancer cells. Akt is a pro-survival kinase that is upregulated in several forms of human cancers. Hyperactive Akt inhibits apoptosis induced by numerous stimuli. However, Akt is unable to inhibit ROS-mediated cell death, and in fact, Akt aids in ROS-directed cell death by inducing cellular oxygen consumption, promoting ROS generation, and impairing ROS degeneration.

Moreover, oxidative stress induced by 3b promoted activation of stress kinases (p38/JNK) and resulted in cancer cell apoptosis. Treatment with antioxidants was able to reduce cell death confirming ROS-mediated cytotoxicity. In conclusion, our novel class of compounds are promising leads that by exacerbating oxygen consumption, ROS production, and reducing the antioxidant capacity of cancer cells tip the balance towards activation of stress kinases that ultimately leads to cell death.

Citation Format: Divya Pathania, Mario Sechi, Michele Palomba, Vanna Sanna, Francesco Berrettini, Angela Sias, Laleh Taheri, Nouri Neamati. Design and discovery of novel small molecule redox modulators as therapies for pancreatic cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4555. doi:10.1158/1538-7445.AM2013-4555
Abstract 4596: LIN28B drives neuroblastoma oncogenesis through let7-MYCN signaling.
Authors: Schulte Johannes H. , Molenaar Jan , Lindner Sven , Schramm Alexander , Mestdagh Pieter , Speleman Frank , Versteeg Rogier , Eggert Angelika
Published: August 2014

LIN28B regulates developmental processes by modulating miRNAs of the let-7 family. A role for LIN28B in cancer has been proposed, but has not been established in vivo. We report LIN28B amplification or overexpression in high risk neuroblastoma. LIN28B repressed let-7 family miRNAs and induced MYCN expression in neuroblastoma cells in vitro. LIN28B-let-7-MYCN signaling induced proliferation and blocked differentiation of normal neuroblasts and neuroblastoma cells. The effect of LIN28B knock-down was rescued by overexpression of let-7-resistant MYCN. To analyse the tumor-initiating capacity and oncogenicity of Lin28b in vivo, mice were generated by knock-in of the CAG-LSL-Lin28b-IRES-Luciferse vector (LSL-Lin28b) into the ROSA26 locus. Crossbreeding these mice with DBH-iCre mice targeted Lin28b expression to the neural crest. Abdominal tumors developed in 7 of 18 DBHiCre;LSL-Lin28b mice at 36-62 days of age. Autopsy revealed uni- or bilateral adrenal tumors in all mice, reflecting the most frequent localization of human NB. Tumors consisted of small round blue cells and expressed the NB markers, DBH, TH and Phox2b. The macroscopic tumor appearance, primary tumor sites, tumor histology and marker gene expression confirmed these tumors as NB. Successful serial transplantation supported that the primary tumors from this model system were fully transformed malignant tumors. Both the Lin28b and MYCN proteins were strongly expressed in all tumors and members of the let-7 miRNA family were significantly downregulated. We demonstrate that overexpressing Lin28b in the neural crest can drive NB tumor formation in mice, supporting LIN28B as an important oncogene for NB and potentially for other malignancies. Therapeutic approaches aimed at inhibiting Lin28b or let-7 family re-expression may be useful therapeutic approaches to circumvent MYCN addiction in NB.

Citation Format: Johannes H. Schulte, Jan Molenaar, Sven Lindner, Alexander Schramm, Pieter Mestdagh, Frank Speleman, Rogier Versteeg, Angelika Eggert. LIN28B drives neuroblastoma oncogenesis through let7-MYCN signaling. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4596. doi:10.1158/1538-7445.AM2013-4596
Abstract 4599: Sequential monitoring of single-cell copy number variation in metastatic prostate cancer.
Authors: Kuhn Peter , Dago Angel E. , Stepansky Asya , Carlsson Anders , Felch Natalie , Luttgen Madelyn , Kolatkar Anand , Hicks James , Gross Mitchell E.
Published: August 2014

The high-definition circulating tumor cell (HD-CTC) assay provides for an enrichment-free approach to CTC identification. Here, we utilized the HD-CTC approach to study androgen receptor (AR) expression combined with single-nucleus sequencing for genome-wide analysis of copy number variation (CNV) in sequential CTCs samples obtained from a patient with metastatic prostate cancer treated with abiraterone acetate (an androgen synthesis inhibitor). At baseline, before initiation of abiraterone treatment, we observed a balanced proportion of AR-negative and AR-positive CTCs. During a brief period of clinical response (marked with a decreased serum PSA and decreased pain) the proportion of AR-positive CTCs declined followed by a rapid increase associated with clinical progression (increased PSA and pain). CNV analysis of single CTCs revealed multiple genomic rearrangements, such as AR amplification along with the chromosomal gains and losses typical of prostate cancer, in multiple cells at baseline. During treatment response, the frequency of CNV alterations significantly declined, followed by a reemergence to a pattern of multiple, complex alterations associated with clinical progression. Detailed analysis of the CNV profiles revealed that many abnormalities were commonly shared between the CTC populations, but a number were unique to the AR+ resistant/hormone refractory CTC population including increased MYC amplification alteration and the AR amplicon that include additional adjacent genes. Remarkably, the reconstruction of tumor lineage history based on the CTC genomic profiles enables us to trace and identify the precise treatment time point where the putative therapy-resistant CTC clone emerges, under therapeutic pressure, until it eventually expanded to become the AR+ resistant CTC population at the point of therapeutic relapse. Overall, our results demonstrated that the integration of the HD-CTC enumeration technology with protein expression and single cell genomic analyses could successfully be applied to real time monitoring of ADT therapy emergent change in a prostate cancer patient, and may provide a direct roadmap for personalized cancer medicine in the near future.

Citation Format: Peter Kuhn, Angel E. Dago, Asya Stepansky, Anders Carlsson, Natalie Felch, Madelyn Luttgen, Anand Kolatkar, James Hicks, Mitchell E. Gross. Sequential monitoring of single-cell copy number variation in metastatic prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4599. doi:10.1158/1538-7445.AM2013-4599
Abstract 461: Inhibition of HMGB1 delays tumor progression, reduces MDSC-mediated immune suppression, and diminishes MDSC-macrophage cross-talk interaction.
Authors: Parker Katherine , Rosenberg Suzanne , Sinha Pratima , Yang Huan , Tracey Kevin , Li Jianhua
Published: August 2014

Cancer immunotherapy aims to harness the power of the immune system to fight advanced stages of cancer. A main hindrance to cancer immunotherapy is tumor-induced immune suppression. Within the tumor microenvironment are a plethora of immune suppressive cells including Myeloid-Derived Suppressor Cells (MDSC). MDSC are present in nearly all types of cancer blocking T cell activation and shifting immunity towards a type 2 tumor-promoting phenotype through cross-talk with macrophages that enhances the secretion of pro-tumor molecules such as IL-10, and simultaneously drops IL-12 production by macrophages. MDSC accumulation is induced by pro-inflammatory cytokines including IL-1β, IL-6 and VEGF as well as alarmins such as S100A8/9. Because the alarmin HMGB1 is increased in many cancers, we are determining if HMGB1 drives MDSC. Inhibition of HMGB1 in vitro by chemical inhibitors Glycyrrhizin and Ethyl Pyruvate decreased cross-talk-induced IL-10 production by MDSC and IL-6 production by macrophages. Ethyl Pyruvate also reduced MDSC suppressive capacity in transgenic T cell suppression assays, suggesting that HMGB1 regulates multiple aspects of MDSC function. Treatment of BALB/c mice with 4T1 mammary carcinoma with HMGB1 inhibitors Glycyrrhizin and Ethyl Pyruvate reduced lung metastases and did not impact primary tumor progression. Treatment of C57BL/6 mice with MC38 colon carcinoma with an HMGB1 neutralizing antibody (2G7) reduced MDSC levels in the blood, spleen, and tumor. HMGB1 consists of two domains, the pro-inflammatory B box domain and an anti-inflammatory A box domain. Treatment of C57BL/6 mice with MC38 tumor with A box significantly reduced primary tumor growth and decreased MDSC levels in the blood. These findings indicate that full-length HMGB1 drives inflammation and tumor progression, while the anti-inflammatory A box domain counteracts full-length HMGB1 and reduces tumor growth and MDSC. These results suggest that HMGB1 drives MDSC accumulation and tumor progression and that A box can be used to target MDSC-mediated tumor-induced immune suppression.

(Research supported by NIH RO1CA115880 and RO1CA84232)

Citation Format: Katherine Parker, Suzanne Rosenberg, Pratima Sinha, Huan Yang, Kevin Tracey, Jianhua Li. Inhibition of HMGB1 delays tumor progression, reduces MDSC-mediated immune suppression, and diminishes MDSC-macrophage cross-talk interaction. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 461. doi:10.1158/1538-7445.AM2013-461
Abstract 462: Soluble CD80 restores T-cell activation and overcomes tumor cell programmed death ligand-1-mediated suppression.
Authors: Haile Samuel , Dalal Sonia P. , Clements Virginia , Tamada Koji , Ostrand-Rosenberg Suzanne
Published: August 2014

Many tumor cells escape anti-tumor immunity through their expression of Programmed Death Ligand 1 (PDL1 or B7-H1), which interacts with T cell-expressed PD1 and results in T cell apoptosis. We previously reported that transfection of human tumor cells with a membrane-bound form of the costimulatory molecule CD80 prevented PD1 binding and restored T cell activation. We now report that membrane-bound CD80 similarly reduces PDL1-PD1-mediated suppression by mouse tumor cells, and that a fusion protein consisting of the extracellular domains of CD80 fused to an Fc domain of IgG1 overcomes PDL1-mediated suppression by human tumor cells. T cell activation experiments assessing costimulation indicate that the soluble CD80 fusion protein mediates its effects by binding to PDL1 and inhibiting PDL1-PD1 interactions. Comparison of the CD80 fusion protein to antibodies specific for PD1 or PDL1 demonstrate that soluble CD80 treatment is more effective in restoring T cell activation than treatment with mAb to either PD1 or PDL1. These studies identify soluble CD80 as an alternative and potentially more efficacious therapeutic agent for overcoming PDL1-induced immune suppression and facilitating tumor-specific immunity.

Citation Format: Samuel Haile, Sonia P. Dalal, Virginia Clements, Koji Tamada, Suzanne Ostrand-Rosenberg. Soluble CD80 restores T-cell activation and overcomes tumor cell programmed death ligand-1-mediated suppression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 462. doi:10.1158/1538-7445.AM2013-462
Abstract 4620: Unconventional genetic determinants of resistance to 5-aza-2’-deoxycytidine.
Authors: Ting Angela H. , Matkin Lars
Published: August 2014

5-aza-2’-deoxycytidine (DAC) is approved by the US Food and Drug Administration for the treatment of myelodysplastic syndrome. It is also being tested in Phase I and II clinical trials for efficacy against other malignancies, including brain, prostate, breast, and lung cancers. Identifying modifier genes that could confer resistance to DAC treatments is, therefore, essential in realizing the full potential of DAC as a chemotherapeutic agent. DAC is a cytosine analog that is incorporated into newly synthesized DNA, where it can prevent propagation of DNA methylation by irreversibly trapping DNA methyltransferases. The consequent reversal of epigenetic silencing of tumor suppressor genes contributes to tumor cell death in the therapeutic context. Previous studies have identified relatively few determinants of DAC resistance, including the equilibrative nucleoside transporters, deoxycytidine kinase, and cytosine deaminase. While these genes are logical candidates because of their functions in cellular pyrimidine uptake and metabolism, our large-scale, unbiased random insertional mutagenesis screen in human cancer cell lines have identified additional, unexpected loci involved in DAC resistance. Our initial screen produced 9 mutant clones resistant to DAC at concentrations of 0.1μM and 1μM. In particular, several of the insertional mutants targeted components of the DNA damage response pathway, including ubiquitin conjugating enzymes and Fanconi anemia complementation group genes, indicating that the primary mechanism of killing by DAC may not be DNA demethylation. This is consistent with the observation that DNA double-strand breaks are among the consequences of DAC treatments. Ongoing functional studies of the mutant clones also suggest that cancer cells with increased efficiency at repairing genomic DNA damage may survive DNA demethylation in the presence of DAC. These results not only inform us of the mechanism of action of DAC-mediated cytotoxicity, our findings also have important clinical implications as these genetic modifiers are also valuable biomarkers and therapeutic targets for achieving personalized cancer treatments involving DAC.

Citation Format: Angela H. Ting, Lars Matkin. Unconventional genetic determinants of resistance to 5-aza-2’-deoxycytidine. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4620. doi:10.1158/1538-7445.AM2013-4620
Abstract 4663: Acquired resistance to gefitinib is associated with cancer stem-like phenotype and EMT in EGFR mutant lung adenocarcinoma.
Authors: Lim Sun Min , Kim Hyun Jung , Ban Young Ho , Ha Sang Jun , Cho Byoung Chul
Published: August 2014

Background: Mutations in the epidermal growth factor receptor (EGFR) gene are associated with increased sensitivity of lung adenocarcinoma to tyrosine kinase inhibitor (TKI) such as gefitinib. During gefitinib treatment, however, the tumor becomes resistant by acquiring specific types of resistance via the somatic changes. Although the underlying mechanisms for acquired resistance include the amplification of c-Met gene and the secondary mutation in EGFR genes, there are still unknown mechanisms that cannot be explained by the two types of genetic changes. Here, we sought to find a novel mechanism to explain TKI-resistance of EGFR-dependent lung adenocarcinoma.

Methods: We developed resistant cells (HCC827-GR) from gefitinib-sensitive HCC827 cells harboring EGFR deletion mutation by chronic exposure to increasing concentrations of gefitinib. Acquired resistance mechanisms of HCC827-GR cells were analyzed.

Results: HCC827-GR showed much less phosphorylation of most receptor kinases including EGFR but higher phosphorylation of downstream signaling molecules, Erk and Akt, than its parental cell. By comparing the gene expression profiles between HCC827 and HCC827-GR, we found that cancer stem cell (CSC) markers, CD44 and ALDH1A1 genes, were highly induced in HCC827-GR compared with those in HCC827. Knockdown of CD44 and ALDH1A1 expression by RNA interference induced inactivation of both Akt and Erk phosphorylation in HCC827-GR. Furthermore, HCC827-GR displayed CSC-like functionality such as colony formation and the phenotype of epithelial-mesenchymal transition (EMT). Higher level of vimentin, ZEB1 and ZFHX4 and lower level of E-cadherin were observed in HCC827-GR together with morphological change. Interestingly, miR200c, a tumor suppressive microRNA, was dramatically decreased in HCC827-GR. Forced expression of miR200c in HCC827-GR abrogated the increased expression of ZEB1 and ALDH1A1, resulting in the restoration of E-cadherin.

Conclusions: These findings imply that cancer stemness induced via CD44 and ALDH1A1 expression and EMT contribute to the acquisition of gefitinib resistance in EGFR-TKI sensitive lung adenocarcinoma. Therefore, CSC markers including CD44 and ALDH1A1 and EMT regulators such as miR200c can be used as therapeutic targets for TKI-resistant EGFR-dependent lung adenocarcinoma.

Citation Format: Sun Min Lim, Hyun Jung Kim, Young Ho Ban, Sang Jun Ha, Byoung Chul Cho. Acquired resistance to gefitinib is associated with cancer stem-like phenotype and EMT in EGFR mutant lung adenocarcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4663. doi:10.1158/1538-7445.AM2013-4663

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 467: Bcl2A1 - an IFN-gamma dependent master switch for the function of CD11b+Gr-1high myeloid derived suppressor cells.
Authors: Haile Lydia , Echeverz Jose Medina , Zhao Fei , Métais Jean-Yves , Dunbar Cynthia , Korangy Firouzeh , Greten Tim
Published: August 2014

Myeloid derived suppressor cells (MDSC) play a crucial role in suppression of immune responses in cancer and inflammation. Here, we describe differential IFN-γ/STAT1 dependent regulation of the suppressor function of CD11b+Gr-1high granulocytic and CD11b+Gr-1dull/int monocytic MDSC subsets. IFN-γ induced STAT1 phosphorylation in both MDSC subsets, which led to NOS2 expression in CD11b+Gr-1dull/int. monocytic MDSC and enhanced suppression of T cell proliferation. In sharp contrast, the suppressor function of CD11b+Gr-1high granulocytic MDSC was enhanced in the absence of IFN-γ using IFN-γR-/- MDSC or IFN-γ-/- effector cells, which led to upregulation of Bcl2a1 and improved survival of the granulocytic subpopulation during MDSC-T cell interaction. Likewise Bcl2a1 overexpression in CD11b+Gr-1high granulocytic MDSC supported their suppressor activity. Thus, depending on the local IFN-γ concentration, STAT1 regulates the function of MDSC subsets antithetically so that these two cell subpopulations can substitute each other and suppress antigen-specific T cell immunity.

Citation Format: Lydia Haile, Jose Medina Echeverz, Fei Zhao, Jean-Yves Métais, Cynthia Dunbar, Firouzeh Korangy, Tim Greten. Bcl2A1 - an IFN-gamma dependent master switch for the function of CD11b+Gr-1high myeloid derived suppressor cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 467. doi:10.1158/1538-7445.AM2013-467
Abstract 4671: Radioguided detection of lymph node metastasis in non-small cell lung cancer.
Authors: Nwogu Chukwumere , Demmy Todd , Yendamuri Sai , Tan Wei , Bogner Paul , Dexter Elisabeth , Picone Anthony , Hennon Mark , Hutson Alan , Reid Mary , Cheney Richard , Adjei Alex
Published: August 2014

Background:

Lymph node staging provides the most important prognostic information in patients with loco-regional non-small cell lung cancer. We hypothesized that gamma radiation detection using an intra-operative hand-held gamma probe following intravenous 18F-fluorodeoxyglucose (FDG) injection would identify lymph nodes containing metastases in a much more sensitive manner than standard pathological practices. We compared the accuracy of detecting thoracic lymph node metastases using positron emission tomography- computed tomography (PET-CT) versus the gamma probe.

Methods:

One hundred patients with resectable lung cancers had pre-operative positron emission tomography-computed tomography (PET-CT) and mediastinoscopy. 10mCi of FDG was injected on the day of surgery. A handheld gamma probe detected increased FDG uptake within thoracic lymph nodes during pulmonary resection procedures. The lymph nodes that demonstrated increased FDG uptake, but were non-malignant by conventional H&E staining underwent further serial sectioning, immunohistochemistry (IHC) and RT-PCR (reverse transcriptase - polymerase chain reaction). Sensitivity and specificity for lymph node metastasis detection by PET-CT and the gamma probe were calculated.

Results:

Three patients had metastatic lymphadenopathy detected at mediastinoscopy, so their procedures were aborted, while the others proceeded to lung resection and complete lymphadenectomy. Fifteen additional patients had lymph node involvement on routine pathologic analysis. IHC and RT-PCR detected micrometastatic lymph node disease in 4 and 29 patients, respectively. Using RT-PCR as the gold standard, the sensitivity and specificity of PET-CT for detection of lymph node metastasis were 11% and 98% respectively, in contrast to 38% and 50% respectively for the gamma probe.

Conclusion:

The intra-operative hand held gamma probe is more sensitive but less specific than PET-CT in detecting lymph node metastasis from lung cancer. Its overall accuracy was low, resulting in limited up-staging of patients. RT-PCR analysis of FDG-avid lymph nodes for epithelial markers increased the clinical utility of this probe in detecting micrometastasis. Such up-staged patients could derive a survival benefit from adjuvant chemotherapy. More specific radioisotopes are being developed to improve the clinical utility of this technique.

Citation Format: Chukwumere Nwogu, Todd Demmy, Sai Yendamuri, Wei Tan, Paul Bogner, Elisabeth Dexter, Anthony Picone, Mark Hennon, Alan Hutson, Mary Reid, Richard Cheney, Alex Adjei. Radioguided detection of lymph node metastasis in non-small cell lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4671. doi:10.1158/1538-7445.AM2013-4671
Abstract 4681: Characterization of pulmonary hypertension in lung cancer.
Authors: Storn Samantha , Pullamsetti Soni Savai , Kojonazarov Baktybek , Schermuly Ralph Theo , Grimminger Friedrich , Rapp Ulf R. , Seeger Werner , Savai Rajkumar
Published: August 2014

Introduction: Approximately 60%-70% of patients with lung cancer have dyspnea at presentation, and 90% experience it just prior to death. Considering case reports describing the formation of pulmonary hypertension (PH) via pulmonary tumor microemboli from gastric and gallbladder cancer, we analyzed the involvement of PH in the development of dyspnea in lung cancer using different mouse models and human lung tumor samples.

Methods: Three murine lung cancer models have been used: C57BL/6N-mice that received an injection of Lewis-lung-carcinoma-cells (LLC1) via tail vein, K-rasLA2 knock-in-mice and SP-C-c-raf-1-BxB-23 transgene mice. They underwent echocardiographic analyses at different time points and hemodynamic measurements to assess the right ventricular systolic pressure (RVSP) and systemic arterial pressure. Histological evaluation was performed to determine the pulmonary vascular morphometry in the tumor-area as well as the total collagen of the right ventricle. Identical histological analyses have been done in human lung tumor samples. In addition, to understand the contribution of immune cells to the development of PH in lung cancer setting, pulmonary vascular walls were analyzed in human lung tumor tissue and quantified for macrophages (CD68+), T-lymphocytes (CD3+) and dendritic cells (CD209+).

Results: For each mouse model, echocardiography reveals an impaired systolic, diastolic and global function of the right ventricle, as well as a dilated right ventricle. Hemodynamics show an increased RVSP. Murine and human lung tumor samples demonstrate vascular remodeling, affecting both wall thickness and the degree of muscularization in the tumor-area. The quantification of total collagen enables to prove a right ventricular fibrosis. Higher incidences of T-lymphocytes and macrophages in pulmonary vascular walls were demonstrated in the tumor-area of human lung section.

Conclusion: This study provides clear evidence for the existence of PH in human as well as animal models affected by lung cancer. Repeated echocardiographic analyses show that the detected PH represents an end-stage phenomenon. The increased content of T-lymphocytes and macrophages in the vascular walls leads us to the hypothesis that these cell types might be involved in the remodeling of small pulmonary vessels in tumors.

Citation Format: Samantha Storn, Soni Savai Pullamsetti, Baktybek Kojonazarov, Ralph Theo Schermuly, Friedrich Grimminger, Ulf R. Rapp, Werner Seeger, Rajkumar Savai. Characterization of pulmonary hypertension in lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4681. doi:10.1158/1538-7445.AM2013-4681
Abstract 4683: Correlation of transcript levels of members of the urokinase-type plasminogen activator system with clinico-pathological parameters in prostate cancer .
Authors: Al-Janabi Omar , Taubert Helge , Wach Sven , Stöhr Robert , Burger Maximilian , Wieland Wolf Ferdinand , Lohse-Fischer Andrea , Fröhner Michael , Wirth Manfred P. , Wullich Bernd , Magdolen Viktor , Kotzsch Matthias , Fuessel Susanne
Published: August 2014

Background: Members of the urokinase-type plasminogen activator (uPA) system play an important role in tumor progression and angiogenesis in different human malignancies including breast, kidney, colon, stomach and prostate cancer (PCa). Aim of the study was to characterize mRNA expression levels of members of the uPA system in tissue specimens from primary PCa and their potential association with clinico-pathological parameters.

Methods: Transcript levels of members of the uPA system, i.e., uPA, uPA receptor (uPAR) and plasminogen activator inhibitor type 1 (PAI-1) were analyzed in a cohort of 132 corresponding pairs of malignant and adjacent non-malignant prostate tissue samples. Relative gene expression levels were determined by quantitative PCR (qPCR) and normalized to the reference gene TATA-box-binding protein (TBP). Potential correlations between the three markers themselves and the clinico-pathological parameters were assessed by Spearman's Rho test and by Fisher's exact test.

Results: There were no significant differences in expression levels of uPA system genes between the corresponding malignant and non-malignant tissue specimens. Transcript levels of all members of the uPA system displayed a significant correlation with each other in the tumor tissues. At stratifying uPA, uPAR and PAI-1 expression levels by the median a significant correlation between an elevated expression of uPA and PAI-1 was correlated with higher Gleason score (P = 0.001 and P = 0.007). In addition, a significant positive correlation between uPAR transcript levels and tumor stage was detected (P = 0.025).

Conclusion: Our results demonstrate that mRNA expression levels of members of the uPA system correlated with each other and with clinico-pathological parameters. We suggest that transcript levels of uPA system members can be helpful markers for the study of tumor behavior of prostate cancer.

Citation Format: Omar Al-Janabi, Helge Taubert, Sven Wach, Robert Stöhr, Maximilian Burger, Wolf Ferdinand Wieland, Andrea Lohse-Fischer, Michael Fröhner, Manfred P. Wirth, Bernd Wullich, Viktor Magdolen, Matthias Kotzsch, Susanne Fuessel. Correlation of transcript levels of members of the urokinase-type plasminogen activator system with clinico-pathological parameters in prostate cancer . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4683. doi:10.1158/1538-7445.AM2013-4683
Abstract 4685: SH2D4A is frequently downregulated in hepatocellular carcinoma.
Authors: Andreozzi Mariacarla , Quagliata Luca , Kovac Michal , Tornillo Luigi , Makowska Zuzanna , Heim Marcus , Heinimann Karl , Piscuoglio Salvatore , Terracciano Luigi Maria
Published: August 2014

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. The lack of effective therapeutic options combined with an increasing incidence rate in the past 20 years calls for the identification of early stage HCC molecular markers. SH2D4A maps to human chromosome 8p21.3 and encodes for SH(2)A. The chromosomal region containing SH2D4A has been shown to be frequently lost in colorectal, lung and HCC cancers. Using a combination of gene expression, protein level and clinical data, our study aimed to investigate the SH2D4A involvement in HCC pathogenesis. Transcriptome analysis (Affymetrix) performed on 37 HCC samples (matched with their corresponding non-neoplastic liver parenchyma) and 5 normal liver donors revealed that SH2D4A is downregulated in HCC. These results were validated by qPCR in fresh frozen tissues [25 out of 37 samples (67.6%) showed SH2D4A downregulation; p<0.026]. Furthermore, the combination of qPCR data and immunohistochemistry staining performed on corresponding formalin-fixed paraffin embedded (FFPE) needle biopsies demonstrated a direct correlation between the SH2D4A mRNA expression and SH(2)A protein levels. Additional data obtained from the analysis of a tissue microarray (TMA) containing 336 informative specimens confirmed that SH(2)A protein levels are frequently reduced in HCC and additionally in cirrhosis nodules compared to normal liver samples. To conclude, our study revealed that SH2D4A is frequently downregulated in HCC samples thus corroborating its putative role as tumor suppressor gene. In addition, we provide new evidence for SH2D4A involvement in HCC pathogenesis demonstrating for the first time its deregulation also in cirrhotic nodules.

Citation Format: Mariacarla Andreozzi, Luca Quagliata, Michal Kovac, Luigi Tornillo, Zuzanna Makowska, Marcus Heim, Karl Heinimann, Salvatore Piscuoglio, Luigi Maria Terracciano. SH2D4A is frequently downregulated in hepatocellular carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4685. doi:10.1158/1538-7445.AM2013-4685

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 4698: Gene expression profiles of triple negative breast cancer epithelial and stromal cells are predictive of treatment response.
Authors: Jovanovic Bojana , Chen Steven X. , Lehmann Brian D. , Johnson Kimberly N. , Sanchez Violeta , Kuba Maria G. , Sanders Melinda E. , Mayer Ingrid A. , Moses Harold L. , Pietenpol Jennifer A.
Published: August 2014

Lacking human epithelial growth factor receptor 2 (HER2) amplification as well as estrogen and progesterone receptors, TNBC is a heterogeneous collection of biologically diverse cancers that likely contributes to variable clinical outcomes for patients suffering from this disease. Since TNBC lacks well-defined molecular targets, we recently performed gene expression (GE) analyses and identified six distinct molecular TNBC subtypes with unique biologies. To further decipher the contribution of tumor epithelium and its adjacent stroma to the biology of TNBC and clinical response to treatments, we performed GE analysis on biopsy material from 20 TNBC patients enrolled in a phase II clinical trial [VICC-BRE0904; a neoadjuvant study of cisplatin and paclitaxel±RAD001 (mTOR inhibitor)]. We isolated RNA from both tumor epithelial and stromal cells using laser capture microdissection. Our goal is to determine if GE from tumor epithelial and/or stromal cells is predictive of response to treatment. Our preliminary results indicate that there is differential GE in both tumor epithelial as well as stromal cells in pretreatment biopsies of responders versus non-responders. Patients with higher expression of proliferation genes such as MKI67, KIFCI, AURKB, E2F3, and DNA damage response genes TP53BP2, CHEK1, RPA1, BLM were more responsive to treatment compared to non-responders. Upon completion of the phase II clinical trial and correlative molecular analyses, we anticipate identifying biomarkers of drug sensitivity that will provide insight to novel combination therapies for the different subtypes of TNBC.

Citation Format: Bojana Jovanovic, Steven X. Chen, Brian D. Lehmann, Kimberly N. Johnson, Violeta Sanchez, Maria G. Kuba, Melinda E. Sanders, Ingrid A. Mayer, Harold L. Moses, Jennifer A. Pietenpol. Gene expression profiles of triple negative breast cancer epithelial and stromal cells are predictive of treatment response. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4698. doi:10.1158/1538-7445.AM2013-4698
Abstract 4706: Wide surgical margins are of prognostic importance in GIST.
Authors: Ahlen Jan , Branstrom Robert , Nilsson Inga-Lena
Published: August 2014

Background: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumor of the gastrointestinal tract and even in the era with molecularly targeted agents is surgery the mainstay of treatment and the only treatment that could lead to cure. Microscopically negative margins are today accepted as sufficient margins for cure while tumor rupture is known to result in poor prognosis and therefore taken into account in prognostic models. In other types of soft tissue sarcoma has wide margins been known to be of importance for recurrence while this has not been shown in GIST.

Methods: 116 patient were included in the study, 55 male and 61 female, mean age 61 years (10-86) and mean follow-up 73 month (4,6-365) that has been evaluated for surgical margins at the time for surgery or at referral to our institution. The criteria used has been those used for sarcoma in other locations were “Wide margins” are defined as 3 cm marginal in the organ of origin combined with an completely intact peritoneal surface covering the tumor. In cases where the tumor is stucked to surrounding tissue or organ, that part has been resected “un bloc” with the tumor. “Marginal margins” are defined as R0 resection with less then 3 cm and “Intralesional margin” is when the tumor is shelled out or damage to surrounding peritoneal surface or judged as R1 resection. Local recurrence has been defined as recurrence at site of origin or intraabdominal recurrence of tumor in the peritoneum and metastatic disease when recurrence is distant metastasis.

Results: We found 48 cases with wide margins(3 recurrence), 39 with marginal margins (15 recurrence) and 29 with intralesional margins (17 recurrence). There was significant difference calculated as Kaplan-Meier for both local recurrence (p=0,00016), distant metastasis (p= 0,004) and survival (p= 0,0047) and in cox regression analysis hazard ratio were 7,4 for marginal margins and 8,08 for intralesional margins compared to wide margins and this was not correlated with size or site (p= 0,007).

Conclusion: Surgical margins is of great importance in GIST both for local recurrence, distant metastasis and survival and patient with GIST should be operated with wide margins, in accordance with principals of other sarcoma.

Citation Format: Jan Ahlen, Robert Branstrom, Inga-Lena Nilsson. Wide surgical margins are of prognostic importance in GIST. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4706. doi:10.1158/1538-7445.AM2013-4706
Abstract 4716: Chronic alcohol consumption activates the LiTAF pathway in melanoma-bearing mice.
Authors: Marron Andrew , Zhang Hui , Zhu Zhaohui , Meadows Gary G.
Published: August 2014

We previously reported that chronic alcohol consumption increases myeloid-derived suppressor cells (MDSC) and compromises melanoma-specific CD8+ T cells in B16BL6 melanoma-bearing mice; however, the underlying mechanism has not been elucidated. Activation of LPS-induced TNF-α factor (LiTAF) gene leads to up-regulation in the expression of various cytokines, chemokines and growth factors such as TNF-α, IL-1β, and VEGF, which are inducers of MDSC. Alcohol consumption is associated with the leakage of LPS into the blood. However, it is not known whether alcohol consumption modulates the expression of LiTAF. Herein, we examined the effects of alcohol and subcutaneous inoculation of melanoma in mice on the expression of LiTAF and its downstream cytokines, chemokines and growth factors. Female C57BL/6 mice were given 20% (w/v) alcohol for three to six months followed by subcutaneous inoculation of 2x105 B16BL6 melanoma cells. Two weeks after tumor inoculation, the expression of LiTAF and related cytokine genes in the spleen were examined by RT-PCR. Chronic alcohol consumption dramatically up regulated the expression of LiTAF in the spleen of melanoma-bearing mice; however, expression was almost undetectable in the spleen of corresponding water-drinking mice. The expression of IL-1β, IL-10, TNF-α, CCL5, and VEGF-β was also significantly up regulated by alcohol. These results indicate that chronic alcohol consumption activates the LiTAF pathway and up regulates the expression of the downstream cytokines, chemokines and growth factors. The up regulation of these cytokines could contribute to the induction of MDSC and inhibition on CD8+ T cells.

(Supported by funds received from the Summer Undergraduate Research Fellowship (SURF) Program, from NIH grants R01 AA07193 and K05AA017149, and from funds provided for medical and biological research by the State of Washington Initiative Measure No. 171).

Citation Format: Andrew Marron, Hui Zhang, Zhaohui Zhu, Gary G. Meadows. Chronic alcohol consumption activates the LiTAF pathway in melanoma-bearing mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4716. doi:10.1158/1538-7445.AM2013-4716
Abstract 4717: Lymphotoxins activate NF-κB-mediated inflammatory and survival pathways in head and neck squamous cell carcinomas.
Authors: Cheng Tsu-Fan , Yang Xinping , Coupar Jamie , Si Han , Lee Steven , Zhang Jialing , van Waes Carter , Chen Zhong
Published: August 2014

Head and neck squamous cell carcinomas (HNSCC) are highly inflammatory and preferentially metastasize to lymph nodes (LN). HNSCC exhibit constitutively activated canonical and alternative NF-κB pathways, but the upstream signals activating these NF-κB pathways have not been well characterized. We hypothesized that immune regulatory cytokines Lymphotoxin (LT) α and β produced by tumor and/or LN cells could be critical in upstream signal activation of NF-κB in HNSCC. Although LTs and their receptors (LTβR and TNFR) are known to be expressed and produced by lymphocytes, we found that LTα and β, and their receptors LTβR and TNFR, are also expressed and produced in a subset of HNSCC cell lines and tumor tissue specimens, when compared with primary human oral mucosal cells. LTα activated both canonical and alternative NF-κB pathways, as indicated by induction of nuclear translocation of NF-κB subunits, RELA, cREL, RELB, p52 in a time dependent manner. In contrast, LTβ mainly activated the alternative NF-κB pathway, enhancing nuclear translocation of RELB and p52. In addition, LTβ-mediated gene expression was examined by Illumina bead microarray platform, and the top pathways of upregulated genes included NF-κB, immunoregulatory, inflammatory, and survival signaling pathways. Our data suggested that HNSCC expressed lymphotoxins and their receptors could be critical factors that contribute to the constitutively activated NF-κB, inflammatory and survival signaling previously implicated in HNSCC pathogenesis. Understanding of lymphotoxin-mediated cross-talk with these inflammatory and survival signaling pathways, could provide a missing link between the inflammation and tumorigenesis in the tumor microenvironment.

Citation Format: Tsu-Fan Cheng, Xinping Yang, Jamie Coupar, Han Si, Steven Lee, Jialing Zhang, Carter van Waes, Zhong Chen. Lymphotoxins activate NF-κB-mediated inflammatory and survival pathways in head and neck squamous cell carcinomas. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4717. doi:10.1158/1538-7445.AM2013-4717