bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5526: Deciphering the basis of CendR-mediated penetration into tumors
Authors: Pang Hongbo , Braun Gary B. , Friman Tomas , Ruoslahti Erkki
Published: August 2015

Cell-penetrating peptides (CPPs) have proven their usefulness in overcoming barriers (such as the cell membrane) of drug delivery into tumor cells. We recently discovered a novel class of cell-penetrating peptides that also have tumor-penetrating properties. These so-called, CendR peptides,bind to neuropilin-1 (NRP1) and initiate an active endocytic process into tumor cells. Subsequent studies showed that CendR-initiated endocytosis is a receptor-dependent macropinocytosis process, and that its activity is stimulated by mTOR inhibition and nutrient deprivation. In vivo, CendR peptides penetrate across tumor vessels deeply into the extravascular tumor tissue, and show ability to improve the tumor accumulation and antitumor efficacy of coupled, and even co-administrated, drugs. Further understanding of the CendR-mediated cell and tissue penetration is of value to cell biology and cancer therapy.

Here, we report ultrastructural studies to unveil the subcellular transport route for CendR peptide. After synchronized cell entry, transmission electron microscopy (TEM) imaging shows that CendR cargo is transported from early endosomes to late endosomes and lysosomes. We have also identified some of the genes important for this transport. Moreover, magnetic isolation of intracellular vesicles containing CendR cargo and proteomics analysis identified signature molecules associated with these vesicles. Studies with TAT peptide comprised of D-amino acids (D-TAT), included in these studies as a non-CendR CPP, indicate that D-TAT also penetrates into tumor cells through a variation of macropinocytosis independent of NRP1. The uniqueness of CendR pathway lies in the linkage of CendR endocytosis to mTOR and cellular nutrient status. We provide evidence that a transcription factor, Sp1, serves as a link between mTOR and CendR endocytosis by regulating the NRP1 level on the cell surface. Additionally, we have shown the intercellular transport of CendR cargo that affords tissue penetration and is also stimulated by nutrient deprivation. Here we identify genes critical to the export and re-entry of CendR cargo during intercellular transport. These results shed light into the mechanism of action of CPPs, and endocytosis in general. They also have implications in cancer metabolism and drug delivery.

Citation Format: Hongbo Pang, Gary B. Braun, Tomas Friman, Erkki Ruoslahti. Deciphering the basis of CendR-mediated penetration into tumors. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5526. doi:10.1158/1538-7445.AM2015-5526
Abstract 5538: A pH-responsive doxorubicin-lipid conjugate loaded nanomedicine for breast cancer treatment
Authors: Li Feng , Saulsbury Marilyn , Heyliger Simone , Bondarev Mikhail , Du Chengan , Ramaley Corinne C. , Lemieux KiTani P.
Published: August 2015

Doxorubicin (DOX) is a chemotherapeutic agent widely used in the treatment of various cancers. However, its clinical use is restricted due to serious cardiotoxicity and the development of drug resistance. Many delivery strategies, including the use of liposomes, micelles, and polymer drug conjugates, have been used to improve the therapeutic efficacy and reduce the toxicity of DOX. Recently, lipid-drug conjugates have also been investigated for the delivery of anticancer drugs. The conjugation of lipids with DOX can increase its lipophilicity, thus enhance its loading in lipid-based nanoparticles, facilitate interaction with cells, and improve pharmacokinetic properties. In this study, we developed a micelle formulation with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-amino polyethylene glycol-2000 (DSPE-PEG) to deliver a DOX-lipid conjugate to treat breast cancers. DOX was conjugated with palmitic acid (PA) through a pH sensitive hydrazone bond to yield a lipophilic pro-drug of DOX (PA-DOX). PA-DOX (Log P = 2) showed significantly increased lipophilicity than DOX (Log P = - 0.5). The micelles were prepared using a thin-film dispersion method. Due to the lipophilic nature of PA-DOX, it can be efficiently loaded into the hydrophobic core of DSPE-PEG micelles. In addition, the micelle formulation demonstrated superior stability, showing no leakage or precipitation of PA-DOX for at least two weeks during storage at 4°C. In vitro drug release was tested in sodium phosphate buffer (pH 7.5) and sodium acetate buffer (pH 5.0), respectively. DOX-PA was stable at pH 7.4 and showed minimal DOX release (less than 10% at 24 hours). The release of DOX was significantly increased at pH 5 (approximately 75% at 24 hours). Anticancer activities were evaluated with MCF-7 breast cancer cells and MCF-7/MDR drug-resistant breast cancer cells. Cell viability was determined by a MTT assay. PA-DOX showed similar anticancer activity on MCF-7 cells as DOX itself. Anticancer activity of PA-DOX was significantly reduced in MCF-7/MDR cells but restored when used in combination with lapatinib.

Citation Format: Feng Li, Marilyn Saulsbury, Simone Heyliger, Mikhail Bondarev, Chengan Du, Corinne C. Ramaley, KiTani P. Lemieux. A pH-responsive doxorubicin-lipid conjugate loaded nanomedicine for breast cancer treatment. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5538. doi:10.1158/1538-7445.AM2015-5538
Abstract 5539: Antiproliferative and apoptotic effects of some African natural products in solid and hematopoietic malignant cell lines
Authors: Akanni Olufemi E. , Berger Martin R.
Published: August 2015

Background: Natural products have played a very important role in health care from time immemorial in curation and prevention of diseases. Antiproliferative and apoptotic effects of extracts of some African natural products like Hymelocardia acida, Moringa oleifera leaves as well as Pleurotus pulmonarius mushroom metabolites were studied in a panel of solid and hematopoietic malignant derived cell lines such as SUIT2-007, MDA-MB 231, SW480, K562 and HL60 cells.

Methods: MTT assay was used to determine the growth inhibition of respective cells in vitro. Apoptosis of the cells was demonstrated by fluorescence microscopy and Hoechst 33342 staining, DNA ladder formation assay and flow cytometry with propidium iodide staining.

Results: MTT results showed that extracts of Hymelocardia acida stem, Moringa oleifera leaves as well as Pleurotus pulmonarius metabolites could inhibit the proliferation of SUIT2-007, MDA-MB 231, SW480, K562 and HL60 cells in a time and dose dependent manner. Our results show that the ethanol extract of Hymelocardia acida stem had a positive effect and induced apoptosis as shown by fluorescence microscopy and flow cytometry assay. Apoptotic effects were demonstrated by nuclear staining with Hoechst showing clear signs for nuclear fragmentation in cells treated with Hymelocardia acida stem and Moringa oleifera leaves extracts. FACS analysis with propidium iodide showed that the sub G1 fraction increased in treated cells. DNA ladder formation, however, showed no clear DNA fragmentation, which does not exclude a clearer picture with isolated compounds.

Conclusion: The present study indicates that the extracts studied can effectively inhibit proliferation and induce apoptosis in cells of panel of tumor cell lines investigated.

Citation Format: Olufemi E. Akanni, Martin R. Berger. Antiproliferative and apoptotic effects of some African natural products in solid and hematopoietic malignant cell lines. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5539. doi:10.1158/1538-7445.AM2015-5539
Abstract 554: Detection and separation of live circulating tumor cells using lipid dye
Authors: Mitra Ranjana , Goodman Jr Oscar , Le Thuc Timothy
Published: August 2015

Purpose: Separation of live circulating tumor cells (CTC) for culturing and monitoring treatment response is highly desirable. We have shown previously that CTC could be distinguished from peripheral blood mononuclear cells (PBMC) based on the intracellular lipid content. This study utilizes intracellular lipid as a biomarker to separate live CTC from PBMC for culturing and assessing treatment response.

Methods: We used Bodipy dye, a fluorescent fatty acid analog that undergoes native transport and metabolism in living cells, to label lipid-rich CTC. LNCaP cells incubated with 50% plasma for 16 hours were used as mock CTC. Test samples contained PBMC (106 cells) spiked with mock CTC (500 cells), which were stained with Bodipy and antibodies against CD45, a biomarker for leukocytes (PBMC). Bodipy+/CD45- CTC were detected and then sorted using a flow cytometer. The sorted Bodipy+/CD45- cells were evaluated for cell viability and their suitability for culturing with trypan blue staining.

Results: The Bodipy/CD45 staining permitted detection and sorting of 375 mock CTC from total 500 spiked LNCAPs (75% efficiency). These cells remained live as confirmed by trypan blue staining indicating suitability for culturing. All sorted cells were further confirmed to be epithelial LNCaP cells with pan-cytokeratin (CK) staining, a biomarker for epithelial cells. The recovery efficiency for live CTC sorting using Bodipy/CD45 staining is comparable with existing methods for isolation of fixed (non-live) CTC using CK/CD45 staining (85%).

Conclusion: Bodipy-positive stain together with CD45 negative stain provided an inexpensive and reliable means to isolate live CTC.

Citation Format: Ranjana Mitra, Oscar Goodman Jr, Thuc Timothy Le. Detection and separation of live circulating tumor cells using lipid dye. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 554. doi:10.1158/1538-7445.AM2015-554
Abstract 5551: Anti-tumorigenic function of polyphenolic compounds on gefitinib resistant lung cancer
Authors: Phan Ai N. H. , Jeong Yangsik
Published: August 2015

EGFR-mutant non-small cell lung cancer (NSCLC) is initially very sensitive to EGFR tyrosine kinase inhibitors (TKIs). Even with the remarkable effect of EGFR TKIs, patients eventually acquire resistance to gefitinib or elortinib, leading to a clinical challenge for overcoming TKI resistance. Some polyphenolic compounds are known as potential anti-tumorigenic compounds, such as resveratrol, curcumin or EGCG. However the function of these natural compounds in resistant NSCLC remains to be elucidated.

Here, we identified a polyphenolic compound (named as compound A) from screening a set of natural compounds for anti-cancer effects on NSCLC. We found that compound A is a potential anticancer agent both in vivo and in vitro. Moreover, the growth inhibition of compound A turns out to be selective for cancer cells. Next, we further evaluated the growth inhibitory effect of compound A in two sets of NSCLC panels which are either sensitive or resistant to gefitinib treatment. Interestingly, the cell lines which are resistant to gefitinib are highly sensitive to compound A treatment, whereas the gefitinib-sensitive (GS) cell lines are not responsive to the compound A treatment. In gefitinib resistant (GR) cells, compound A increases expression of autophagy markers, p62 and LC3, which might result in the growth inhibition of the GR panel of cells but not in the GS cell panel.

Moreover, we profiled the metabolic genes in the GR and GS cell lines. Surprisingly, compound A treatment significantly reduces the expression of a subset of enzymes associated with Warburg effect, which includes GLUT1, LDHA, PDK1, HK1, and HK2. As a result, compound A decreases Warburg effect, subsequently suppressing cell proliferation, especially of the GR cell lines.

Taken together, compound A decreases Warburg effect and up-regulates autophagy process that may contribute to tumor suppression function of compound A in gefitinib-resistant cancer.

Citation Format: Ai N. H. Phan, Yangsik Jeong. Anti-tumorigenic function of polyphenolic compounds on gefitinib resistant lung cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5551. doi:10.1158/1538-7445.AM2015-5551
Abstract 5559: Novel synthetic pyridyl analogues of CDDO-Im with improved stability and their potential use in cancer prevention
Authors: Cao Martine , Onyango Evans , Williams Charlotte , Royce Darlene , Gribble Gordon , Sporn Michael , Liby Karen
Published: August 2015

Synthetic oleanane triterpenoids are non-cytotoxic, multifunctional drugs with a broad spectrum of applications for prevention and treatment of cancer and for many other chronic diseases. CDDO-Im, 1[2-Cyano-3,12-dioxooleana-1,9(11-dien-28-oyl] imidazole, synthesized more than a decade ago, is one of the most potent triterpenoids known to date with marked anti-inflammatory, cytoprotective, antiproliferative, differentiative and proapoptotic activity on various human and murine tumor cell lines. However, pharmacokinetics of CDDO-Im are not optimal. Therefore, three new pyridyl analogues of CDDO-Im, namely CDDO-3P-Im, CDDO-2P-Im and CDDO-4P-Im, have been synthesized and screened for their possible use as chemopreventive or chemotherapeutic drugs. At low nanomolar concentrations, they were equivalent to CDDO-Im for induction of differentiation in U937 leukemia cells and at higher doses they induced apoptosis. As inflammation and oxidative stress contribute to carcinogenesis, we also assessed their cytoprotective potential. The new compounds suppressed inducible nitric oxide synthase expression in RAW264.7 macrophage-like cells and significantly induced heme oxygenase-1 and NADPH quinone reductase mRNA and protein levels in lung cancer cells as well as in various mouse tissues. Most importantly, pharmacokinetic studies performed in vitro in human plasma and in vivo revealed superior stability for each new analogue. While CDDO-Im was almost completely degraded after 30 min (< 12% of starting material remaining) in human plasma, the new compounds were more stable with > 50% still detectable. Six hours after gavage, much higher concentrations of the new derivatives were found in mouse liver, lung, pancreas and kidney in contrast to CDDO-Im. Thus, the new pyridyl analogues have better bioavailability, and because of their potent anti-inflammatory activity and improved stability, they will be tested in vivo in relevant carcinogenesis models.

Citation Format: Martine Cao, Evans Onyango, Charlotte Williams, Darlene Royce, Gordon Gribble, Michael Sporn, Karen Liby. Novel synthetic pyridyl analogues of CDDO-Im with improved stability and their potential use in cancer prevention. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5559. doi:10.1158/1538-7445.AM2015-5559
Abstract 5561: Violacein inhibits matrix metalloproteinase mediated CXCR4 expression: Potential anti-tumor effect in cancer invasion and metastasis
Authors: Tiriveedhi Venkataswarup
Published: August 2015

Matrix metalloproteinases (MMP-2 and -9) play an important role in the tumor metastasis through cleavage of proinflammatory chemokines. Violacein a small molecule naturally produced by Chromobacterium violaceum has been implicated with anti-tumor effects. In this study we investigated the molecular basis of violacein mediated downregulation of CXCL12-CXCR4, chemokine-receptor expression. Gelatin zymography analysis demonstrated that violacein significantly inhibited the cytokine (TNFα and TGFβ) mediated MMP-2 activation in MCF-7 breast cancer cell line. MMP-2 plays a critical role in the secretion of inflammatory CXCL12 a cell migration chemokine. ELISA analysis demonstrated that violacein induced significantly decreased secretion of CXCL12 from cytokine induced MCF-7 cells. Further, we show that MMP-2 and MMP-9 act synergistically at two distinct steps towards the membrane expression of the tumor metastasis chemokine receptor, CXCR4. Violacein efficiently downregulated the CXCR4 membrane expression through MMP-9 inhibition. Taken together, these studies demonstrate the unique mechanism of action of violacein towards potential anti-tumor effect through reduction of CXCL12-CXCR4 expression by inhibition of proteolytic MMP activity. Based on these studies violacein could offer a novel therapeutic venue in cancer therapy.

Citation Format: Venkataswarup Tiriveedhi. Violacein inhibits matrix metalloproteinase mediated CXCR4 expression: Potential anti-tumor effect in cancer invasion and metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5561. doi:10.1158/1538-7445.AM2015-5561
Abstract 557: A novel integrated i-FISH technology to detect and characterize non-hematopoietic heteroploid CTC subtypes with or without expressing vimentin or other tumor biomarkers
Authors: Lin Peter
Published: August 2015

Most of the current circulating tumor cell (CTC) detection technologies rely on expression of EpCAM and cytokeratins (CK) as well as the size of tumor cells shed into peripheral blood of cancer patients or tumor animal models. However, it has been demonstrated that expression of EpCAM and/or cytokeratins in CTCs might be down-regulated or absent during epithelial-mesenchymal transition (EMT), potentially resulting in significant false negative detection of CTCs. Whereas expression of vimentin in EMT tumor cells has been reported. In this study, a new strategy integrating phenotypic immunofluorescent staining of vimentin and karyotypic FISH has been well developed to detect and characterize CTC and its subtypes isolated by subtraction enrichment in various biofluid samples including blood, bone marrow, malignant pleural effusion and ascites as well as cerebrospinal fluid of cancer patients or blood samples of mice bearing human tumors. Both large and small and clusters of vimentin positive or negative non-hematopoietic CTCs with heteroploid chromosome(s) were identified. The similar procedure has also been successfully extended to other desired single or combined tumor biomarkers in addition to vimentin as needed. The established SET-iFISH® platform will help identify and characterize diversified CTC subtypes which may possess distinct clinical significance in terms of drug sensitivity, resistance, and tumor metastasis as well as recurrence, respectively.

Citation Format: Peter Lin. A novel integrated i-FISH technology to detect and characterize non-hematopoietic heteroploid CTC subtypes with or without expressing vimentin or other tumor biomarkers. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 557. doi:10.1158/1538-7445.AM2015-557
Abstract 5571: Technologies for molecular epidemiology in cancer
Authors: Divi Rao L. , Verma Mukesh , Dickherber Anthony
Published: August 2015

Future success in molecular epidemiology depends on how quickly epidemiology adopts, optimizes and validates emerging technologies for molecular-scale analysis. In order to advance molecular epidemiology in cancer, the National Cancer Institute (NCI) supports the development of novel, next-generation, and cutting-edge molecular and cellular analysis technologies. Here we present past and present technology development efforts and their impact on cancer epidemiology. Some of the technologies, whose early-stage development was supported by the NCI, such as resequencing arrays, gene expression assay system, technologies which enable next generation sequencing, RNA preservative, rolling circle amplification of DNA, digital PCR system, functionalization of quantum dots, chromatin immunoprecipitation with next gen Sequencing, and multidimensional protein identification technology are now mainstay in basic and clinical research laboratories for genomic, epigenomic, transcriptomic and proteomic analysis. Additionally, these technologies have significantly contributed in identifying molecular determinants of cancer, and in assessing risk, disease progression and prognosis in large scale cancer epidemiology. Current awards support developing technologies for single cell analysis which can capture heterogeneity, simultaneous detection of multiple oncogenes, sensitive sequencing of geneomic/somatic mutations, targeted sequencing, rare variant detection, sequencing of ultralow-frequency mutations, epigenetic changes, kinome, multiplexed detection of microRNA, proteins, cytokines and metabolites, circulating tumor cells, and highthroughput protein-DNA interactions assessment. In addition, several novel biospecimen collection, preservation and processing technologies are also being supported with the potential to define the future of molecular epidemiology. Collectively, these technology development efforts supported by NCI provide a tremendous opportunity for molecular epidemiologists wishing to study archived samples collected in large-scale epidemiology efforts, and optimize and validate them for new molecular analyses.

Citation Format: Rao L. Divi, Mukesh Verma, Anthony Dickherber. Technologies for molecular epidemiology in cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5571. doi:10.1158/1538-7445.AM2015-5571
Abstract 5580: Characterization of HPV integration sites in the human genome
Authors: Bodelon Clara , Untereiner Micahel , Vinokurova Svetlana , Machiela Mitchell J. , Wentzensen Nicolas
Published: August 2015

Background. Persistent infection with carcinogenic human papillomaviruses (HPV) is known to be cause of the majority of anogenital cancers as well as some head and neck cancers. With the progression of the disease, the HPV genome is commonly integrated within the host DNA, although the mechanism by which integration may promote the progression of the disease is not well-understood. We performed a systematic review of integration events in HPV-related cancers and analyzed the genomic regions where these events occur.

Methods. A PubMed search was performed to collect all published data on integration loci. A total of 29 articles were eligible for the analysis. Determination of genomic integration sites for papers that provided exact nucleotide sequences of integration sites or viral fusion regions was done by using the Basic Local Alignment Search Tool (BLAST, http://blast.ncbi.nlm.nih.gov/Blast.cgi) for the GRCh37/hg19 build. We used the Encyclopedia of DNA Elements (ENCODE) data to investigate if certain genetic features clustered with integrations sites. The Wilcoxon rank sum test was computed to compare the observed and expected distributions of minimal distances from the integration sites with known nucleotide positions to the different genetic features.

Results. As of November 2014, approximately 1,200 integration events were reported in the literature from both tumors and cell lines. The majority of integrations sites (53%) were reported in cervical tumors, 9% in head and neck tumors, 4% of oropharyngeal tumors and 23% in cell lines,. Six percent and 2% of the integration sites were found in high-grade and low-grade cervical lesions, respectively. The majority of reported integration events (77%) were related to HPV16. We found that integration sites from tumors were closer to known genes (p <10−14) and CpG islands (p <10−9) than expected by chance. Among repetitive elements, long terminal repeats (LTR) were enriched in integration sites (P = 0.001). Open chromatin based on the FAIRE-seq peaks showed significant enrichment in integration sites (p <10−11).

Conclusions. In summary, integration sites occur in genomic regions with enrichment for genetic elements, such as gene-rich regions, CpG sites, repetitive elements and open chromatin regions. This corroborates that accessible regions of the genome are preferentially targeted by integration and suggests that host genes may be functionally altered in development of HPV-related cancers.

Citation Format: Clara Bodelon, Micahel Untereiner, Svetlana Vinokurova, Mitchell J. Machiela, Nicolas Wentzensen. Characterization of HPV integration sites in the human genome. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5580. doi:10.1158/1538-7445.AM2015-5580
Abstract 5584: Exhaled breath as diagnostic tool for malignant pleural mesothelioma
Authors: Lamote Kevin , Lardon Filip , Van Cleemput Joris , Nackaerts Kristiaan , Thas Olivier , van Meerbeeck Jan P.
Published: August 2015

Background: Malignant Pleural Mesothelioma (MPM) is an asbestos-related disease with a dismal prognosis due to its late detection at an advanced stage. Hence, a diagnostic test is requisite that allows to screen asbestos-exposed persons for earlier MPM detection. As blood biomarkers have not shown to be clinically useful biomarkers for early non-invasive diagnosis, breath is currently explored. Breath is easy to retrieve by physicians in a clinical environment and contains volatile organic compounds (VOCs) that arise from (patho)physiological processes. Since asbestos causes oxidative stress and cancers are known to up regulate their metabolism, we hypothesize that VOCs and, hence, the exhaled breath of MPM patients will differ from healthy controls.

Methods: We compared the breath of 23 MPM patients, 10 asbestos-exposed and 12 non-exposed healthy individuals using a multicapillary column/ion mobility spectrometer (MCC/IMS, B&S Analytik, Dortmund, Germany). After subjects refrained from eating, drinking and smoking for at least 2 hours, 10 ml alveolar air was sampled via a CO2-controlled ultrasonic sensor and subsequently analyzed. Per subject a background sample was taken. Eighty-nine VOC peaks were visually selected via on-board VisualNow 3.7 software. Their intensity was compared between background and breath samples. After calculating the VOCs’ alveolar gradient, we performed a logistic LASSO regression in R (R-Foundation for Statistical Computing, Vienna, Austria). We selected the optimal MPM diagnostic logistic LASSO model by 10-fold cross-validation using age, gender and the alveolar gradient of the peaks and estimated this models’ sensitivity, specificity, AUCROC and positive (PPV) and negative predictive value (NPV).

Results: We were able to discriminate MPM patients from the asbestos-exposed and non-exposed controls with 96% sensitivity and 91% specificity. The AUCROC was 0,94 and the PPV and NPV were resp. 92% and 95%. The VOCs P5, P3, and P1 were the significant discriminators.

Conclusions: Breath analysis in a clinical setting by MCC/IMS allows to discriminate MPM patients from asbestos-exposed and non-exposed healthy controls. It can help to screen asbestos-exposed persons at risk, switching diagnosis to an early phase and hence, better patient survival. Identification of the underlying VOCs and a further validation in different patient cohorts is being undertaken.

Citation Format: Kevin Lamote, Filip Lardon, Joris Van Cleemput, Kristiaan Nackaerts, Olivier Thas, Jan P. van Meerbeeck. Exhaled breath as diagnostic tool for malignant pleural mesothelioma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5584. doi:10.1158/1538-7445.AM2015-5584
Abstract 5586: Breast cancer risk prediction: An update to the Rosner-Colditz breast cancer incidence model
Authors: Rice Megan , Tworoger Shelley , Hankinson Susan , Tamimi Rulla , Eliassen A Heather , Colditz Graham , Rosner Bernard
Published: August 2015

Purpose: To update and build on the Rosner-Colditz breast cancer incidence model by evaluating the contributions to breast cancer risk of childhood and adolescent somatotype, fruit and vegetable intake, breastfeeding, and physical activity. Secondarily, we evaluated the contributions of these variables to breast cancer incidence models defined by ER and PR status of the tumor.

Methods: Using data from the Nurses’ Health Study (NHS) and Nurses’ Health Study II (NHSII), we added childhood somatotype (average of somatotype at ages 5 and 10; 9 unit scale), adolescent somatotype (average of somatotype at ages 10 and 20; 9 unit scale), fruit and vegetable intake (servings/day), breastfeeding (months), and physical activity (METs/week) to the Rosner-Colditz breast cancer incidence model to determine whether these variables improved the discriminatory power of the model. We evaluated the contribution of these variables to a model for all invasive breast cancer as well as to models for ER+/PR+, ER+/PR-, and ER-/PR- breast cancer.

Results: In the NHS/NHSII, we accrued over 3000 cases of invasive breast cancer with complete data on the selected breast cancer risk factors. Compared to the original Rosner-Colditz model, the models that included childhood or adolescent somatotype were significantly improved (p<0.001). The relative risk (RR) of invasive breast cancer for a 4 unit change in somatotype was 0.69 for childhood somatotype and 0.59 for adolescent somatotype. Including childhood somatotype increased the age-adjusted concordance statistic from 0.613 to 0.616 whereas including adolescent somatotype increased the age-adjusted concordance statistic from 0.613 to 0.618. Childhood and adolescent somatotypes also significantly improved ER+/PR+ and ER-/PR- breast cancer incidence models (p<0.02). Breastfeeding and vegetable intake suggestively improved the ER-/PR- model. The RR for five servings of vegetables per day was 0.82, while the RR for 12 months of breastfeeding was 0.87. While these variables were significantly associated with ER-/PR- breast cancer risk in the updated models, the differences in concordance statistics comparing models with to models without these variables were not statistically significant.

Conclusion: Childhood and adolescent somatotype significantly improved breast cancer risk classification in the NHS/NHSII.

Citation Format: Megan Rice, Shelley Tworoger, Susan Hankinson, Rulla Tamimi, A Heather Eliassen, Graham Colditz, Bernard Rosner. Breast cancer risk prediction: An update to the Rosner-Colditz breast cancer incidence model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5586. doi:10.1158/1538-7445.AM2015-5586
Abstract 5588: Nevus count and dermoscopic pattern associated with MC1R RHC-variant alleles in a case-control study of melanoma
Authors: McClenahan Phil , Jagirdar Kasturee , Lee Katie , McEniery Elizabeth , Beh Sam , Burke Brian , Duffy David L. , Soyer H. Peter , Sturm Richard A.
Published: August 2015

The total number of acquired melanocytic nevi (TNC) is the strongest known risk factor for cutaneous malignant melanoma (CMM). An increased risk of CMM is also seen with individuals with red hair, fair skin and poor tanning ability (RHC phenotype); and who carry variant alleles of the MC1R gene designated as R. High freckling scores but reduced nevus counts have been reported in individuals with the RHC phenotype. The relationship between these reciprocal phenotypes of lower vs higher nevus counts, both with increased risk of CMM, is complex and supports hypotheses of different pathways to melanoma formation. We have reported on a case-control study of nevi in volunteers from the South-east Queensland area combining an assessment of pigmentation characteristics, nevus phenotype including dermoscopic nevus subtypes with genotypic comparisons and melanoma risk for 573 individuals. This sample set has been further examined to delineate the nevus characteristics of 46 individuals who are homozygous MC1R RHC allele carriers R/R. We wished to determine if there is any commonality in the pigmentation or melanocytic nevus phenotype including number, size, distribution, profile and colour that may distinguish these patients. Body mass index (BMI), body surface area (BSA), TNC and MC1R genotype were analysed and found that participants with personal melanoma history had higher TNC (P<0.005). We found no association between MC1R genotype and BMI but there exists a weak association between BSA and higher TNC. Within the MC1R R/R cohort, melanoma patients are more likely to have high TNC than controls (P<0.01). The signature dermoscopic pattern for the individuals in the MC1R R/R group is at increased odds of being a non-specific pattern (P<0.01). A significant difference in MC1R R/R TNC dependent on melanoma history exists and there is a predisposition towards a non-specific signature nevus pattern.

Citation Format: Phil McClenahan, Kasturee Jagirdar, Katie Lee, Elizabeth McEniery, Sam Beh, Brian Burke, David L. Duffy, H. Peter Soyer, Richard A. Sturm. Nevus count and dermoscopic pattern associated with MC1R RHC-variant alleles in a case-control study of melanoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5588. doi:10.1158/1538-7445.AM2015-5588
Abstract 5589: Age-stratified risk of unexpected uterine sarcoma following surgery for presumed benign leiomyoma
Authors: Brohl Andrew S. , Li Li , Andikyan Vaagn , Cioffi Angela , Dudley Joel T. , Kasarskis Andrew , Maki Robert G.
Published: August 2015

Background: Estimates of unexpected uterine sarcoma following surgery for presumed benign leiomyoma that utilize age-stratification are lacking.

Patients and Methods: A retrospective cohort of 2075 patients that had undergone myomectomy was evaluated to determine the case incidence of unexpected uterine sarcoma. An aggregate risk estimate was generated using a meta-analysis of similar studies plus our cohort. Database-derived age distributions of the incidence rates of uterine sarcoma and uterine leiomyoma surgery were used to stratify risk by age.

Results: Of 2075 patients in our cohort, six were diagnosed with uterine sarcoma. Our meta-analysis revealed 8 studies from 1980-2014. Combined with our cohort, there are 18 cases of leiomyosarcoma reported in 10120 patients, for an aggregate risk of 1.78 per 1000 [95% CI: 1.1-2.8], or 1 in 562. There are 8 cases of other uterine sarcomas reported in 6889 patients, for an aggregate risk of 1.16 per 1000 [95% CI: 0.5-4.9], or 1 in 861. The summation of these risks gives an overall risk of uterine sarcoma of 2.94 per 1000 [95% CI: 1.8-4.1], or 1 in 340. After stratification by age, we predict the risk of uterine sarcoma to range from a peak of 10.1 cases per 1000, or 1 in 98, for age 75-79 to less than 1 case per 500 for age <30.

Conclusion: The risk of unexpected uterine sarcoma varies significantly across age groups. Our age-stratified predictive model should be incorporated to more accurately counsel patients and to assist in providing guidelines for surgical technique for leiomyoma.

Age-stratified risk of unexpected uterine sarcoma at the time of surgery for presumed leiomyomaAgeEstimated(years)Risk of Uterine Sarcoma (per 1000 procedures)25-291.74(1 in 574)30-342.01(1 in 496)35-392.65(1 in 377)40-442.46(1 in 405)45-493.28(1 in 304)50-544.62(1 in 216)55-596.35(1 in 158)60-646.36(1 in 157)65-697.53(1 in 133)70-748.55(1 in 117)75-7910.1(1 in 98)80-845.57(1 in 179)

Citation Format: Andrew S. Brohl, Li Li, Vaagn Andikyan, Angela Cioffi, Joel T. Dudley, Andrew Kasarskis, Robert G. Maki. Age-stratified risk of unexpected uterine sarcoma following surgery for presumed benign leiomyoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5589. doi:10.1158/1538-7445.AM2015-5589
Abstract 5590: Reliability of medical records in diagnosing inflammatory breast cancer in Egypt
Authors: Le Lynne , Schairer Catherine , Hablas Ahmed , Ramadan Mohamed , Merajver Sofia , Seifeldein Ibrahim , Soliman Amr
Published: August 2015

Inflammatory breast cancer (IBC) is a rare and aggressive form of breast cancer characterized by diffuse erythema and edema (peau d’orange) of the skin of the breast. It does not include neglected locally advanced cancer of the breast presenting late in the course of the disease. IBC is primarily a clinical diagnosis, but also requires pathologic confirmation of breast cancer. The purpose of this study was to examine the reliability of medical records for diagnosing IBC in patients at two centers in Tanta, Egypt (Gharbiah Cancer Society and Tanta Cancer Center) which participated in an epidemiologic case-case study of IBC and non-IBC in 2009-2010. IBC patients were recruited to the case-case study at diagnosis based on clinical examination; detailed information on the extent and duration of signs used for the diagnosis of IBC was recorded. Information on signs of IBC similar to those collected in the case-case study was abstracted from the medical records of the hospitals where case-case study patients were recruited during the same time period, then linked and compared to the signs observed from clinical examination for the case-case study.

A total of 40 IBC cases were recruited into the case-case study during 2009-2010. Medical record information was available for 31 of these cases. Twenty-eight of these 31 cases were reported to have erythema and 28 edema/peau d’orange based on the clinical examination for the case-case study. Of those with erythema, 16 (57.1%) also had erythema/inflammation listed in the medical record; of those with edema/peau d’orange, 23 (82.1%) had this mentioned in the medical record. Erythema was noted on the medical record but not recorded in the case-case study for 2 cases; there were no cases in which edema/peau d’orange was present in the medical record and not in the case-case study. Our analysis indicates that medical records at these cancer centers often have missing information regarding the presence of signs that are necessary to the diagnosis of IBC. Overall, this study suggests that medical record review is not sufficient to identify IBC cases for an epidemiologic study.

Citation Format: Lynne Le, Catherine Schairer, Ahmed Hablas, Mohamed Ramadan, Sofia Merajver, Ibrahim Seifeldein, Amr Soliman. Reliability of medical records in diagnosing inflammatory breast cancer in Egypt. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5590. doi:10.1158/1538-7445.AM2015-5590
Abstract 56: Receptor-Interacting protein kinase 1 functions as an oncogenic regulator in human melanoma
Authors: Jin Lei , Liu Xiao Ying , Lai Fritz , Yan Xu Guang , Jiang Chen Chen , Guo Su Tang , Wang Chun Yan , Croft Amanda , Tseng Hsin-Yi , Wilmott James S. , Scolyer Richard A. , Zhang Xu Dong
Published: August 2015

Although many studies in recent years have uncovered an important role of receptor-interacting protein kinase 1 (RIP1) in mediating cell survival and death signaling, its potential part in the pathogenesis of cancer remains less understood. Here we report that RIP1 functions as an oncogenic regulator in human melanoma. While the expression of RIP1 was commonly upregulated in melanoma, knockdown of RIP1 inhibited melanoma cell proliferation in vitro, and retarded melanoma growth in a xenograft model. Conversely, despite induction of apoptosis in a small proportion of melanoma cells, overexpression of RIP1 enhanced proliferation in the remaining cells. The promoting effect of RIP1 on melanoma cell proliferation was mediated by activation of NF-κB, as blockade of NF-κB activation eliminated RIP1 overexpression-triggered increase in cell proliferation, whereas hyperactivation of NF-κB abolished inhibition of cell proliferation caused by RIP1 knockdown. In support, RIP1 knockdown led to reduction, whereas its overexpression caused an increase, in NF-κB activation. Strikingly, ectopic expression of RIP1 enhanced melanocyte proliferation and triggered anchorage-independent growth of the cells similarly in a NF-κB-dependent manner. While upregulation of RIP1 was associated with DNA copy number gain in a subset of melanomas, constitutive ubiquitination and subsequent stabilization of the RIP1 protein driven by TNFα autocrine appeared to be another mechanism commonly responsible for upregulation of RIP1 in melanoma cells. Collectively, these results identify RIP1 as an oncogenic regulator in melanoma, and points to the possibility of targeting the NF-κB activating mechanism of RIP1 as a novel approach in the treatment of the disease.

Citation Format: Lei Jin, Xiao Ying Liu, Fritz Lai, Xu Guang Yan, Chen Chen Jiang, Su Tang Guo, Chun Yan Wang, Amanda Croft, Hsin-Yi Tseng, James S. Wilmott, Richard A. Scolyer, Xu Dong Zhang. Receptor-Interacting protein kinase 1 functions as an oncogenic regulator in human melanoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 56. doi:10.1158/1538-7445.AM2015-56
Abstract 560: Retrospective evaluation, of the randomized Phase 3 MARQUEE trial of tivantinib (T) + erlotinib (E) versus placebo (P) + erlotinib (E) using VeriStrat in patients with previously treated nonsquamous NSCLC
Authors: Scagliotti Giorgio , Akerley Wallace , von Pawel Joachim , Roder Joanna , Shuster Dale , Schwartz Brian , Spinella Dominic
Published: August 2015

Background: MET, the high-affinity receptor for hepatocyte growth factor, is frequently deregulated in human cancer and is thought to be involved in resistance to EGFR TKIs. Tivantinib is an orally administered MET inhibitor currently under development as a cancer therapy. The Phase 3 MARQUEE trial enrolled 1048 subjects with locally-advanced or metastatic non-squamous NSCLC who had received one or two lines of prior systemic therapy. Patients were randomized 1:1 to T+E (N = 526) or P+E (N = 522) until disease progression. In the ITT population, addition of tivantinib to erlotinib significantly improved PFS and ORR but did not improve OS, the primary endpoint. VeriStrat® (VS) is a multivariate serum protein test that utilizes MALDI-TOF mass spectrometry to assign a “GOOD” (VS-G) or “POOR” (VS-P) classification and has demonstrated broad prognostic and predictive utility for EGFR TKI treatment regimens.

Methods: Pre-treatment serum samples from the MARQUEE study were blinded to clinical outcome and analyzed by VeriStrat. Clinical benefit of T+E vs. P+E was analyzed as a function of VS status, in the ITT population as well as in pre-defined subgroups.

Results: A total of 996 subjects were classified: 718 as VS-G and 278 as VS-P. Subjects testing VS-G had superior outcomes to those testing VS-P in both the T+E arm (OS HR = 0.32, CI: 0.26-0.41, p<0.0001; PFS HR = 0.55, CI: 0.44-0.69, p<0.0001) and P+E arm (OS HR = 0.43, CI: 0.34-0.55, p<0.0001; PFS HR = 0.70, CI: 0.55-0.88, p<0.028). VeriStrat status was a significant, independent predictor of OS, in these erlotinib treated patients, after adjustment for EGFR mutation, KRAS mutation, or MET IHC status. However, VeriStrat did not predict OS or PFS benefit for T+E vs. P+E (treatment * VS interaction p-value for OS of p = 0.09 and PFS of p = 0.19). VS was neither predictive nor prognostic for tivantinib effect in any molecular subgroup.

Conclusion: These data support the previously demonstrated ability of VS to be both prognostic and predictive for survival in erlotinib treated patients, but not for tivantinib treated patients.

Citation Format: Giorgio Scagliotti, Wallace Akerley, Joachim von Pawel, Joanna Roder, Dale Shuster, Brian Schwartz, Dominic Spinella. Retrospective evaluation, of the randomized Phase 3 MARQUEE trial of tivantinib (T) + erlotinib (E) versus placebo (P) + erlotinib (E) using VeriStrat in patients with previously treated nonsquamous NSCLC. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 560. doi:10.1158/1538-7445.AM2015-560
Abstract 561: Development of a highly specific Met antibody for screening Met overexpression in NSCLC
Authors: Chen Caiwei , Lu Hsiangmin Emily , Wei Haitao , Yuan Kehu , Ma Donghui , He Wei-Wu
Published: August 2015

Approximately 5%-22% of non-small cell lung cancer (NSCLC) patients with secondary resistance to EGFR TKIs had evidence of amplification of the Met oncogene. This dysregulation of Met signaling was implicated in the proliferation and eventual metastasis of NSCLC, and therefore represents an attractive target for anticancer drug development.

Currently, there are a number of clinical trials for Met targeted small molecule and monoclonal antibody therapies. Immunohistochemistry (IHC) analysis of the Met protein will be very important for these Met targeted therapies and requires a highly specific antibody to prevent false positives. To develop a highly specific monoclonal antibody against Met for IHC analysis in NSCLC, we generated 97 monoclonal antibodies. IHC evaluation showed that 8 of the antibodies had suitable performance in Met positive NSCLC patient tissues. The 8 Met monoclonal antibodies were further tested on a high density protein microarray chip containing 17,000 human proteins. The microarray analysis showed that clone 3G5 was highly specific as evidenced with it only reacting with Met on the microarray. This new 3G5 Met monoclonal antibody will be a promising candidate as a diagnostic reagent for detecting Met protein in NSCLC with minimal false positives.

Citation Format: Caiwei Chen, Hsiangmin Emily Lu, Haitao Wei, Kehu Yuan, Donghui Ma, Wei-Wu He. Development of a highly specific Met antibody for screening Met overexpression in NSCLC. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 561. doi:10.1158/1538-7445.AM2015-561