bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4915: Physiologically-based PK modeling of liposomal drug delivery points to a key role of tumor deposition in determining the relative efficacy of liposomal vs. free doxorubicin in breast cancer and Kaposi sarcoma
Authors: Hendriks Bart , Reynolds Joe , Klinz Stephan , Leonard Shannon , Geretti Elena , Lee Helen , Eckelhofer Isabelle , Gaddy Daniel , Wickham Thomas
Published: June 2012

For the chemotherapeutic, doxorubicin, liposomal encapsulation simultaneously (i) protects the heart from doxorubicin-induced cardiotoxicity, (ii) results in passive tumor accumulation via the enhanced permeability and retention effect and (iii) increases its circulation life-time. Clinically, pegylated-liposomal doxorubicin (PLD) shows a dramatic benefit to Kaposi sarcoma (KS) patients relative to free doxorubicin. In contrast, PLD does not show any survival benefit relative to free doxorubicin in patients with metastatic breast cancer. We hypothesized that the differential efficacy observed might be explained by differences in tumor cell exposure to drug. To better understand the effects of liposomal encapsulation on drug exposure, we developed a mechanism-based PK model of liposome biodistribution. The model consists of superimposed PK models for free and liposomal doxorubicin with a common central blood compartment which are then connected to a physiologically-based tumor tissue compartment. The tumor tissue compartment includes vascular, interstitial and cell space and captures rates of blood flow, tumor deposition into the interstitial space, doxorubicin release from the liposomes and the trafficking of free doxorubicin into and out of cells. The model parameters were extensively validated against literature data from in vitro studies, mouse xenografts and human clinical data. With our model we are able to simulate total tumor doxorubicin and DNA-bound doxorubicin in tumors, reproducing observed mouse xenograft data for free vs. liposomal doxorubicin. Analysis of the model revealed dramatically different dynamics in tumor exposure for free vs. liposomal doxorubicin delivery. Further, rates of tumor deposition were rate-limiting for overall tumor cell exposure for liposomally-delivered doxorubicin. Upon scaling the model to reflect human physiology, we demonstrate that the clinically observed liposome accumulation in KS tumors can quantitatively explain the dramatic increase in efficacy relative to free doxorubicin. Furthermore, the model indicates that the lower rates of liposome tumor deposition seen in breast cancer patients result in no survival benefit relative to free doxorubicin, consistent with clinical trial results.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4915. doi:10.1158/1538-7445.AM2011-4915
Abstract 4922: A single-cell TGFbetaR-JUND dichotomy and its role in basal-like breast cancer
Authors: Wang Chun-Chao , Jamal Leen , Atkins Kristen A. , Janes Kevin A.
Published: June 2012

Basal-like carcinoma is a subtype of breast cancer with poor prognosis that is characterized by pronounced cell-to-cell non-uniformities. Intratumor heterogeneity could involve sporadic single-cell activation of key pathways implicated in cancer progression, but it has not been possible to identify such pathways in an unbiased way. Our lab has developed a new approach for globally identifying coregulated, heterogeneously expressed genes among single cells (Nat Methods 7:311 [2010]). In a 3D in vitro culture model of a basal-like breast epithelial cell line (MCF10A), we stochastically profiled 4557 transcripts for heterogeneity and identified 547 that were predicted to be strongly dichotomous among extracellular matrix (ECM)-attached cells in the developing acinus. Hierarchical clustering of the heterogeneous transcripts revealed a group of ∼15 biosynthetic genes that contained the transcription factor, JUND, and the basal-cell cytokeratin, KRT5. In the same dataset, we identified a second group of genes-containing a transforming growth factor-beta (TGF-beta) family ligand (GDF11), a TGF-beta receptor (TGFBR3), and a TGF-beta marker protein (TGFBI)-with strong sampling fluctuations that were opposite to those of JUND cluster. Here, we show that perturbation of JUND or TGFBR3 expression profoundly disrupts 3D morphogenesis. Constitutive JUND expression causes stable cribiform-like bridges across the acinar lumen, whereas TGFBR3 knockdown induces a ductal-branching phenotype. These perturbations further revealed that JUND and TGFBR3 are reciprocally coupled by interlinked positive and negative feedback loops that could give rise to asynchronous oscillations during morphogenesis. Live-cell imaging with a fluorescently tagged Smad2 revealed sporadic pulses of TGF-betaR signaling in matrix-attached cells during morphogenesis. Last, in clinical specimens of breast cancer with dichotomous basal cytokeratin expression, we find that junD and Krt5 proteins are correlated in ECM-attached cells. Interestingly, junD and Krt5 are anticorrelated in cells not directly in contact with stromal ECM, and this regulation is mimicked in 3D culture of MCF10A cells. Together, our results raise the possibility that heterogeneous basal-like breast cancers switch between two states that depend critically on the ECM context.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4922. doi:10.1158/1538-7445.AM2011-4922
Abstract 4924: Gene network inference models reveal novel key mediators of mammary tumorigenesis
Authors: Brock Amy , Li Hu , Krause Silva , Collins James J. , Ingber Donald E.
Published: June 2012

Cancer is a developmental disease, requiring the concerted switching of multiple cellular gene programs. Here we ask whether we can use computational network inference models in a systems biology-driven approach to identify changes in connectivity within the underlying gene regulatory network during cancer progression, as well as novel key mediator genes that are central to altered disease state networks. To investigate this hypothesis, we reverse engineered the gene regulatory networks of mammary glands within normal mice and transgenic C3(1)-SV40Tag mice that undergo mammary gland transformation and tumor progression. RNA was isolated from the mammary glands of transgenic mice at 8, 12, 16 and 20 weeks of age; wild type mice were analyzed at 8 weeks. Whole genome microarray profiles were obtained for each stage. A network inference algorithm was trained on a set of 3000 genome-wide gene expression signatures from public databases. A Mode-of-action Network Inference (MNI) algorithm was used to identify key mediator genes at each stage of the gene regulatory network during different stages of cancer progression in the transgenic mouse mammary gland based on z-score significance. From this initial set of candidates, we focused on key mediator genes classified by gene ontology analysis as transcription factors. To experimentally assess the functional relevance of genes identified by this computational approach, key mediator genes were silenced in vitro in three-dimensional (3D) cultures of mouse mammary tumor epithelial cells derived from the transgenic animals. In contrast to normal mammary epithelial cells, tumor cells failed to form polarized structures containing a lumen in 3D spheroid culture. Importantly, siRNA knockdown of key mediator genes induced these cancer cells to revert to a normalized phenotype in 3D culture. These data demonstrate the feasibility of using genome reverse engineering approaches to identify genes that can potentially induce cancer reversal.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4924. doi:10.1158/1538-7445.AM2011-4924
Abstract 4946: Aberrant methylations of microRNA-9s in lung cancers
Authors: Muraoka Takayuki , Toyooka Shinichi , Sou Junichi , Maki Yuho , Ueno Tsuyoshi , Tanaka Norimitsu , Aokage Keiju , Asano Hiroaki , Tsukuda Kazunori , Yamane Masaomi , Takahiro Oto , Miyoshi Shinichiro
Published: June 2012

Background: microRNAs (miRNAs) represent a new class of small non-coding RNAs regulating gene expression by inducing messenger RNA degradation or interfering with translation. miRNAs regulate many cellular processes and have been shown to be associated with cancer development. Recent study showed that miR-9s consisting of mir-9-1,9-2, 9-3 regulates E-cadherin. On the other hand, it has been reported that that methylation of miR-9s are associated with metastasis or poor prognosis of some type of human cancers. In this study, we examined the methylation status of miR -9-1, 9-2, 9-3 in lung cancer to find that miR-9 methylation seems to be associated with lymph node metastasis in lung cancer(LC).

Material and methods: We collected 152 LC tumor DNAs which resected in our institute during 2007 to 2008. After bisulfate conversion, we determined the methylation status of miR-9-1, 9-2 and 9-3 by combined bisulfite restriction analysis method.

Results: Aberrant methylations of either miR-9 were shown in 52 out of 152 cases (34%). Methylation of either miR-9 was present in 39 %, 26 %, 34 %, 42 % of males, females, never-smokers, and ever-smokers, respectively. Methylation of either miR-9s were detected in 35 % of adenocarcinomas, 33 % of squamous cell carcinomas, and 63 % of other LCs. Of note, miR-9 methylation was significantly more frequent in pathological stage I cases (39 %, n = 119) than in the other stage (II to IV) cases (18 %, n = 33; p = 0.028). While 4 cases were methylated in lymph node metastasis cases (17%, n =24), 48 cases were methylated in cases without lymph nodal metastasis (38 %, n =128 : p=0.06).

Conclusion: Aberrant hypermetylation of miR-9s were associated with early pathological stage and /or non-lymph nodal metastasis, suggesting that miR-9s, which relate to aggressiveness and metastasis, might be sliced by abbarent methylation in early stage LCs.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4946. doi:10.1158/1538-7445.AM2011-4946
Abstract 4949: microRNA (miR) analysis as a tool for discovering platinum resistance mechanisms in non-small cell lung cancer (NSCLC)
Authors: Bar Jair , Perez-Iratxeta Carolina , Gorn-hundermann Ivan , Stefanie Reid , Goss Glenwood D. , Dimitroulakos Jim
Published: June 2012

Background: NSCLC is the most common cause of cancer-related death in the world. Platinum-based chemotherapy is the mainstay of treatment both in the adjuvant and the metastatic settings, but platinum resistance is common. A variety of mechanisms have been reported to underlie platinum-resistance. miRs are short non-coding RNA molecules, each regulating a large number of genes. We hypothesized that in platinum-resistant NSCLC cells, a small number of differentially expressed miRs modulating an abundance of gene products, activate in concert a large number of platinum-resistance mechanisms.

Aim: To uncover mechanisms of platinum-resistance in NSCLC, by indentifying miRs differentially expressed in platinum resistant cells.

Methods: Platinum-resistant cell lines were created by cisplatin selection of Calu6 and NCI-H23, NSCLC cell lines. RNA was hybridized to Affymetrix miR arrays, followed by TargetScan algorithm analysis. Levels of specific miRs were quantified by RT-PCR (ABI) and were modulated by miR inhibitors (Ambion).

Results: Two sets of sensitive (S) and platinum-resistant (R) cell lines were created, each set consisting of a parental S line and several R sub-lines. miR array analysis was performed on the S and R lines. miRs that were significantly more abundant in each of four R lines compared to the corresponding S lines were compiled. Genes predicted to be targeted by each of those miRs with a p value < .05 were tabulated. Surprisingly, only 16 genes were common to those four gene lists, including CDK6, an important G1-S cell cycle regulator. mRNA and protein levels of CDK6 were higher in S compared to R lines, and accordingly, growth rate of R lines was slower. miR145, one of the miRs upregulated in the R lines, is predicted to target CDK6. Transfection of a miR145 inhibitor resulted in a mild increase in mRNA levels of CDK6, supporting its role as a regulator of CDK6.

Conclusions: Utilizing a number of biological systems (sets of cells lines) that model the same phenotype (platinum resistance) allows for a robust analysis of the involved molecules. miR profiling of multiple systems can identify genes that are commonly regulated in different systems by different miRs. Using this approach we discovered upregulation of miRs, including miR145, resulting in downregulation of CDK6, as a potential mechanism of platinum resistance in NSCLC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4949. doi:10.1158/1538-7445.AM2011-4949
Abstract 4963: An integrated approach for functional analysis of 11q13 amplification target genes in HNSCC
Authors: Lepikhova Tatiana , Pehkonen Henna , Louhimo Riku , Chen Ping , Peitsaro Nina , Nieminen Anni , Imai Misa , Klefstrom Juha , Hautaniemi Sampsa , Monni Outi M.
Published: June 2012

Gene amplification is one of the most common mechanisms for oncogenic activation in human cancer. Some genomic regions are more frequently amplified than others. For example, amplification of 11q13 occurs at high frequencies in several human cancers, such as lung, bladder, breast and ovarian carcinomas, as well as head and neck squamous cell carcinoma (HNSCC). Approximately 40% of HNSCC are affected by this specific amplification, which is associated with the presence of lymph node metastasis and poor prognosis. Distinct core regions of amplified areas within the 11q13 locus have been identified, and these can be amplified independently or concurrently in various combinations. A number of oncogenes or potential cancer-related genes have been mapped to the 11q13 chromosomal region, however it is still not known whether these genes, that undergo co-amplification and over-expression, cooperate in promoting tumorigenesis. We have used a complementary approach to comprehensively characterize the functional basis for selection and maintenance of 11q13 amplification in HNSCC. To facilitate this approach, each of the 15 HNSCC cell lines used in this study has undergone molecular characterization for genomic and transcriptomic alterations. We performed a loss-of-function screen on 11q13 positive HNSCC cell lines to identify the genes essential for growth and survival. Next we applied gain-of-function analysis using ORFs for CTTN, FADD, PPFIA1, ORAOV1, CCND1 on 11q13 negative HNSCC cell lines. A single or multiple genes mapping to the amplicon were stably co-overexpressed in different combinations and related phenotypes were analysed in several in vitro and in vivo assays for growth, invasion and sensitivity/resistance to drugs. In addition, changes associated with overexpression of those genes were analysed by exon arrays in order to identify deregulated signaling pathways associated with 11q13 amplification. Our preliminary data suggests that this integrated approach is an efficient method at identifying genes that co-operate due to their location in the same amplicon. Such information is particularly valuable for elucidating oncogenic drivers and addressing the relevance of 11q13 amplification to drug sensitivity/resistance in 11q13 positive HNSCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4963. doi:10.1158/1538-7445.AM2011-4963
Abstract 4987: Identification of novel genes and pathways involved in resistance to HER2-targeting agents in breast cancer
Authors: Wetterskog Daniel , Shiu Kai-Keen , Chong Irene , Meijer Titia , Natrajan Rachael , Lord Christopher J. , Ashworth Alan , Reis-Filho Jorge S.
Published: June 2012

The HER2 gene ERBB2 is amplified and/ or overexpressed in up to 15% of all invasive breast cancers. These cancers are characterised by an aggressive phenotype and poor clinical outcome. HER2 positive tumours often display multiple high level amplifications of genes in addition to HER2. Although targeted therapies to HER2 have proven clinical benefit, a substantial number of patients have tumours that are either de novo resistant or acquire resistance over time to these agents. Reasons for this are still largely unknown. By using a high throughput siRNA screen we sought to determine whether genes recurrently amplified and overexpressed in HER2-amplified breast cancer mediate resistance to HER2-targeting agents. We profiled 45 HER2-amplified primary breast cancers and 13 HER2-amplified cell lines with high resolution microarray-based comparative genomic hybridisation (aCGH) and Illumina WG6 v2 arrays. Overlaying of aCGH and gene expression data led to the identification of 369 genes recurrently amplified and overexpressed when amplified. Lapatinib-resistant HER2-amplified breast cancer cell lines were treated with lapatinib, a HER2 small molecule inhibitor, and screened with an RNA interference (RNAi) library targeting these 369 genes and controls. This RNAi screen and subsequent validation screens identified eight genes that when silenced lead to a significant sensitization to lapatinib. The mechanisms of resistance conferred by the identified genes were further investigated and pointed to the involvement of NF-kappaB-signalling, Wnt-signalling and reactive oxygen species scavenging. Our results indicate that specific amplified and overexpressed genes found in HER2 positive breast cancers mediate resistance to anti-HER2 agents. These findings might be used to identify patients likely to be de novo resistant to lapatinib and provide new targets for combined targeted therapies to overcome resistance to anti-HER2 therapies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4987. doi:10.1158/1538-7445.AM2011-4987
Abstract 4995: Lung specific target gene inhibition following intravenous delivery of siRNA nanocomplexes
Authors: Fewell Jason , Sparks Jeff , Congo Richard , Slobodkin Gregory , Rea-Ramsey Angela , Pence Casey , Matar Majed , McClure Diane , Rice Jennifer , Brunhoeber Elaine , Wilkinson Leslie , Anwer Khursheed , Polach Kevin
Published: June 2012

RNA interference offers the promise of a new class of therapeutics that can specifically target many diseases that have aberrant gene expression as the root cause. This is potentially true even for diseases that are currently considered non-druggable by small molecules or other conventional therapies. The means to efficiently deliver RNAi based therapeutics into target cells still poses arguably the most significant barrier to overcome for this platform technology. Cationic systems (lipids and polymers) have been extensively used for siRNA delivery and a wide range of experimental approaches have indicated that siRNA specific knockdown in vivo can be achieved using these systems. However, there is considerable concern that the dose levels required to achieve therapeutically relevant knockdown will result in unacceptable toxicity. This is particularly relevant when applied to systemic delivery. To address this issue we have developed a novel core cationic lipopolyamine structure that is chemically flexible allowing for incorporation of active moieties that improve safety, stability, and efficiency, creating a family of functionalized RNAi delivery systems. Here we show that systemic (intravenous) administration of siRNA nanocomplexes formulated with one of these functionalized systems produces significant target gene knockdown specifically in the lung with duration of at least 7 days following a single injection. Administration was well tolerated with no indications of acute or long term toxicities. Detailed biodistribution analysis reveals that siRNA is initially distributed primarily to lungs, liver and spleen, with the majority of the siRNA accumulating in the liver. Lung specific knockdown is explained through the observed rates of siRNA clearance, which are considerably faster for the liver and spleen than those observed in the lung. This results in higher lung siRNA concentrations on the timescale of RNAi mediated transcript knockdown. In a model of metastatic lung cancer administration of siRNA targeting STAT3 resulted in significant transcript depletion in the lungs and a significant reduction in tumor burden. These results support the continued evaluation of this delivery system as a potential siRNA based therapeutic approach for a variety of diseases of the lung.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4995. doi:10.1158/1538-7445.AM2011-4995
Abstract 5002: Posttranslational modification of E2F1 during melanoma progression
Authors: Meng Peng , Ghosh Rita
Published: June 2012

Melanoma, the malignant tumor of melanocytes, accounts for approximately 78% of skin cancer related deaths. No current treatment significantly enhances disease-free survival once distant metastases have established. To facilitate the development of effective treatment strategies, it is imperative that we obtain a better understanding of the biology of melanoma cells as they progress through the various stages of disease. Previous studies have shown that E2F1, a transcription factor, is over-expressed in melanoma cells. Interaction between E2Fs and tumor suppressor, Rb is essential for regulation of cell cycle progression. E2F1 is known to be involved in other important biological processes, including DNA damage, DNA repair, differentiation, development, autophagy and apoptosis. However, the mechanism through which E2F1 is elevated in melanoma remains unknown. In normal cells in late S phase, E2F1 is recognized by the E3 ligase p45 SKP2, undergoes ubiquitination and is finally degraded in 26S proteasome. Since multiple post-translational modifications have been reported to affect the stability and functions of E2F1, we have hypothesized that during melanoma progression E2F1 fails to undergo degradation and this failure is due to post-translational modification of E2F1. In this study we have used melanoma cells of increasing disease progression. Our results show that primary melanoma cells have high level of E2F1 message and protein. We also found that E2F1 protein levels are not tightly regulated during cell cycle progression. Levels of E2F1 remain constantly high during cell cycle progression in the less metastatic melanoma cells. Consistent with these results, phosphorylation of E2F1 at Serine 337 is almost unchanged during cell cycle progression in the less metastatic melanoma cells. These results taken together suggest that this post-translational modification site may play an important role in stabilization of E2F1 in the primary melanoma cells that represent the radial and vertical growth phases of melanoma. Since modifications usually induce conformational changes, which may affect any following modifications, it is necessary to study the crosstalk among different modifications like acetylation and phosphorylation. The modifications towards the same direction may generate a specific array of modifications that can induce markedly stronger functional effects compared with those produced by the sum of single modifications. Therefore, it is intriguing to detect the coordination of different modifications and determine if there is such a modification array that regulates the stabilization and degradation of E2F1 resulting in the elevated E2F1 levels in melanoma cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5002. doi:10.1158/1538-7445.AM2011-5002
Abstract 5017: Comprehensive analyses of the T cell-mediated adaptive immune function in the elderly melanoma patients
Authors: Hegde Upendra P. , Yadav Meeta , Chakraborty Nitya G.
Published: June 2012

Introduction: The incidence of melanoma is rising at a rapid rate in elderly population. Melanoma is an immunogenic tumor but T lymphocyte in the elderly persons are believed to be hypo/dysfunctional. Methods: HLA-A2 positive eleven elderly and eight young patients (median age 82 Y and 48 Y respectively) participated in the IRB approved study following informed consent. Peripheral blood (PB) derived CD8+ cytotoxic T lymphocytes (CTL), CD4+ helper T lymphocytes (TL) and CD4+CD25+ natural regulatory (nTreg) were used. Mart-1 and flu peptide was presented through PB derived antigen presenting cells (APC) to respective autologous CD8+ T cells to generate antigen specific CTL. The respective CD8+ and CD4+ cells from six elderly and 5 young patients, were engineered to express MHC class I restricted Mart-1 specific TCR and tested in functional assays. nT reg cells were tested for suppressive effect upon anti-CD3 mediated proliferation of autologous CD4+CD25- cells. Results: We found no significant differences in transduction efficiencies of Mart-1 specific TCR between young and old patients. Both Mart-1 and Flu specific CTL could be generated from all the 11 elderly and eight young patients. In vitro expanded antigen specific T effector cell responses were also comparable with inherent and engineered TCR between the two age groups. The elderly group, however, exhibited a significantly lower suppressive nTreg activity (94.4 % in young vs.16.7% suppression in elderly of CD4+TL stimulated by anti CD3 antibody). Conclusion: Our studies show that the CTL function is comparable between the two groups of patients but the nTreg activity in elderly patients is significantly reduced. More critical studies will be needed in larger number of patients to develop a comprehensive understanding of the T cell-mediated function that is important for immune based treatment of elderly melanoma patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5017. doi:10.1158/1538-7445.AM2011-5017
Abstract 5021: Physical health scores and breast cancer outcomes in the ABC Pooling Project
Authors: Pierce John P. , Flatt Shirley W. , Natarajan Loki , Shu Xiao O. , Nechuta Sarah , Lu Wei , Caan Bette , Chen Wendy Y.
Published: June 2012

Background: Of the two health-related quality of life domains measured by the SF-36 (mental health and physical health), the physical health score (PHS) has been associated with all cause mortality, and recently with additional breast cancer events. This PHS includes subscales for self-perceived physical functioning, general health, bodily pain, and role limitations due to physical problems and is known to be a marker for a number of co-morbidities. Methods: Three of the four cohorts that form the After Breast Cancer (ABC) Pooling Project measured health-related quality of life using the SF-36 after cancer diagnosis. These cohorts were the Women's Healthy Eating and Living (WHEL) Study, the Nurses’ Health Study (NHS), and the Shanghai Breast Cancer Survival Study (SBCSS). Using Cox proportional hazards models controlled for cancer stage and grade, we examined the relationship between PHS with disease-free and overall survival in 9387 early stage (I-III) breast cancer survivors. The post-diagnosis SF-36 measurement took place between 1991 and 2006 and the median follow-up time was 7.8 years. Poor physical health was defined as a non-normalized score of 76.4 as previously published by the WHEL study. We also report the association between PH and co-morbidities. Results: The pooled HR (95% CI) for additional breast cancer events was 1.27(1.12, 1.44) with significant findings in each of the two US cohorts: Nurses’ Health Study 1.31(1.07-1.61) and WHEL study 1.28(1.08-1.51). A similar effect size was seen in SBCSS only, with the measurement taken during or immediately after cancer treatment. However, mostly likely due to the much shorter follow-up in this study, it did not reach statistical significance (HR=1.22 95% CI 0.80-1.86). In all cohorts, low PHS was strongly associated with higher BMI. In overweight/obese WHEL participants, low compared to adequate PHS scores were significantly associated with much lower levels of physical activity (p<.0001), 64% higher reported insomnia, a 50% higher prevalence of hypertension and diabetes, and a twofold higher prevalence of arthritis. Conclusion: The importance of the PHS as a predictor of future breast cancer events and overall survival appears to be confirmed in these observational studies. Interventions to improve the PHS and prognosis might target weight control, physical activity and sleep management.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5021. doi:10.1158/1538-7445.AM2011-5021
Abstract 5055: The role of estrogens and vascular endothelial growth factor (VEGF) in premenopausal breast cancer carginogenesis and disease progression
Authors: Recchia Francesco , Candeloro Giampiero , Desideri Giovambattista , Necozione Stefano , Rea Silvio
Published: June 2012

Background. Estrogen is important in breast cancer development and there is evidence in vitro, that it modulates VEGF expression in breast cancer cells through transcriptional activation. However few clinical studies have investigated the association between VEGF and estrogens in humans. Objective of this study was to evaluate weather estrogen suppression with an LH-RH analogue was able to downregulate VEGF expression, and thus decrease its plasma concentration, in premenopausal patients with high-risk early breast cancer.

Patients and methods: From April 2003 to October 2008 we conducted this study on 100 premenopausal early breast cancer patients. At baseline, after surgery, before any type of therapy, plasma VEGF and sex hormones (estradiol, progesterone, FSH, LH) were evaluated. Evaluation was repeated each year. Patients received an LH-RH analogue for 5 years, a chemotherapy that was tailored to the biological characteristics of each patient, radiation therapy, and hormonal therapy in estrogen receptor positive (ER+) tumors. Primary end point was the evaluation of VEGF. Secondary end points were progression-free survival (PFS) and overall survival (OS).

Results: Median age was 43 years (range 26-45); mean number of positive axillary nodes was 3.3. (Range 0-25). Fifty-three patients were ER+ and progesterone receptor positive (PGR+), 17 were ER negative (−) and PGR-, 30 were ER+ and PGR-, Median KI-67 was 33% (range 15%-100%). Twenty patients were Herb-2 positive by FISH. A statistically significant decrease of VEGF was observed after 1 year. At 5 years and during the study period, plasma VEGF continued to decrease (p<0.001) in 94% of patients that were disease-free, while in 6% of patients with disease relapse, VEGF increased with respect to baseline values. No unexpected toxicity of chemotherapy was observed, while hot flashes and G1 osteopenia occurred after LH-RH analogues administration. After a median follow-up of 50 months (range 24-90), 8-year disease free survival and overall survival rated were 94% and 100%, respectively.

Conclusion: Estrogen deprivation with an LH-RH analogue is able to decrease plasma VEGF levels in premenopausal high risk breast cancer patients. These data show how estrogens, through VEGF modulation, may be responsible for the worst prognosis that is observed in premenopausal breast cancer patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5055. doi:10.1158/1538-7445.AM2011-5055
Abstract 5097: Proteomic characterization of non-small cell lung cancer (NSCLC): Discovery and validation of NSCLC protein signatures and signaling networks
Authors: Wei Yuhong , Taylor Paul , Tong Jiefei , Strumpf Dan , Ignatchenko Vladimir , Pham Nhu An , Thyparambil Sheeno , Tsao Ming-Sound , Kislinger Thomas , Moran Michael F.
Published: June 2012

Lung cancer is the leading cause of cancer death worldwide. Non-small cell lung carcinoma (NSCLC) accounts for 80% of lung cancers. The most prevalent subtypes of NSCLC are adenocarcinoma (ADC) and squamous cell carcinoma (SCC), which combined account for approximately 90%. Ten resected NSCLC patient tumors (5 ADC and 5 SCC) were directly introduced into severely immune deficient (NOD-SCID) mice, and the resulting xenograft tumors (XT) were analyzed by standard histology and immunohistochemistry (IHC) and by proteomics profiling. Mass spectrometry (MS) methods involving 1- and 2-dimensional LC-MS/MS, and multiplexed selective reaction monitoring (mSRM, or MRM), were applied to identify and quantify the xenograft proteomes. Hierarchical clustering of protein profiles distinguished between the ADC and SCC subtypes. The differential expression of >175 proteins was found to constitute a distinctive proteomic signature associated with NSCLC subtype, and an mSRM method was developed that provided relative quantification of a subset of highly differentially expressed proteins (i.e. >10-fold; p <0.05) that distinguished ADC and SCC subtypes. The mSRM assay was further developed for the absolute quantification of tumor signature proteins, and also the epidermal growth factor receptor (EGFR) and phosphorylated EGFR, and was successfully applied with formalin-fixed paraffin-embedded tissue sections following laser micro dissection and Liquid Tissue® processing (Expression Pathology, Rockville, MD). To further validate the XT models and to define tumor-specific protein signatures, proteomic profiles encompassing ∼1500 proteins were compared between 5 (each) resected NSCLC patient tumors (PT), corresponding XT, and normal lung tissue (N). Subsets of proteins were identified that, based on their quantified differential expression patterns, distinguished between N and PT, and validated the fidelity of tumor signatures in the XT models. Protein quantifications by mSRM were associated with superior dynamic range and reproducibility but were otherwise consistent with orthogonal methods including IHC and Western immuno blotting. These findings illustrate the potential to develop a comprehensive MS-based platform in oncologic pathology for better classification and potentially treatment of NSCLC patients.

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Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5097. doi:10.1158/1538-7445.AM2011-5097
Abstract 510: Do adipocytes in the tumor microenvironment influence the growth of pancreatic cancer
Authors: White Patrick , Swartz-Basile Deborah , Ziegler Kathryn , Wang Sue , Pitt Henry , Zyromski Nicholas
Published: June 2012

Introduction: Pancreatic cancer develops more frequently and progresses more rapidly in obesity. The mechanisms influencing this association are incompletely understood. Using our murine model of pancreatic cancer in diet-induced obesity, we hypothesized that changes in the immune profile and tumor microenvironment may contribute to accelerated pancreatic cancer growth in obesity.

Methods: Thirty male C57BL/6J mice were studied. At 5 weeks of age, 20 mice were fed high fat diet (60% fat; HFD) and 10 were fed low fat (10% fat) diet. At 19 weeks of age all mice were inoculated in the flank with 2.5 x105 Pan02 murine pancreatic cancer cells. After 5 weeks of tumor growth, spleens and tumors were collected, splenic flow cytometry evaluated lymphocyte population, proliferation was determined by PCNA staining, and tumor infiltrating lymphocytes (TIL), both B and T, were scored by immunohistochemistry. Intratumoral adipocyte volume was assessed by H&E staining. Students's t-test and Pearson's correlation were applied where appropriate. P value <0.05 was accepted as statistically significant.

Results: Mice were segregated into overweight (OW – heavier than the mean body weight of the HFD mice – 35.5g, n=15) and lean (<35.5g, n=14). OW mice were significantly heavier (37.3±0.2g vs. 33.9±0.3g, p<0.001). Tumors were twice as large in OW mice as in lean mice (1.23±0.2g vs. 0.6±0.1g; p=0.001). The peripheral lymphocyte profile was similar in both OW and lean animals (T cells: 51.2±2.5% vs. 49.2±2.3%, p=0.59; B cells: 32.9±1.0% vs. 32.0±1.9%, p=0.73). TIL were observed in similar numbers in both OW and lean groups (T cell: 1.31±0.18 vs. 1.54±0.16, p=0.35; B cell: 0.63±0.08 vs. 0.91±0.15, p=0.33). Tumor proliferation as measured by PCNA was similar in both groups (139±18 OW vs. 143±14 lean, p=0.85). Interestingly, adipocyte volume was significantly greater in the OW tumor microenvironment than in the lean tumors (3.7%±0.7 vs. 2.2%±0.3, p<0.05).

Conclusions: These results demonstrate that: 1) tumor weight was significantly greater in OW mice; 2) peripheral and tumor infiltrating lymphocyte profile was similar in OW and lean animals; and 3) adipocyte volume was significantly greater in the tumor microenvironment of OW mice. We conclude that obesity accelerates the growth of pancreatic cancer, and adipocytes in the tumor microenvironment may directly influence tumor growth.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 510. doi:10.1158/1538-7445.AM2011-510
Abstract 5111: Integrative phospho-proteomic and genomic analyses identify AXL as a potential biomarker and therapeutic target for NRAS-mutated melanoma
Authors: Fedorenko Inna V. , Paraiso Kim H.T. , Joseph Beulah D. , Fang Bin , Koomen John , Eschrich Steven , Nathanson Katherine L. , Wubbenhorst Bradly , Mathew Rahel , Messina Jane , Smalley Keiran S.M.
Published: June 2012

Currently, few therapeutic options exist for the 15-20% of patients whose melanomas harbor activating mutations in NRAS. Here, we have used comprehensive phospho-proteomic and genomic analysis to characterize the patterns of intracellular signaling in a panel of NRAS and BRAF-mutated melanoma cell lines, with the goal of identifying new biomarkers and therapeutic targets for NRAS mutated melanomas. Our preliminary analyses showed BRAF mutated melanomas to have less diverse intracellular signaling than the NRAS group, which tended to have constitutive phosphorylation in a wide-range of receptor tyrosine kinases (RTKs), such as c-MET, EGFR, HER2 and c-Abl. Although these patterns of RTK activity were heterogeneous across the NRAS mutated melanoma cell line panel, all of the NRAS mutated cell lines examined showed constitutive activity in the receptor tyrosine kinase Axl. A role for Axl in the oncogenic behavior of NRAS mutated melanomas was suggested by array CGH studies showing genomic amplification of Axl in a significant fraction of the NRAS mutated melanoma cell lines and by Western Blot experiments demonstrating increased expression of Axl and its ligand Gas6. Mass spectrometry analysis and immunoprecipitation revealed Axl to be constitutively phosphorylated at the Tyr702 autophosphorylation site in NRAS mutated melanoma cell lines only. Functionally, siRNA knockdown and pharmacological inhibition of Axl impaired the survival and invasion of NRAS mutated melanoma cell lines grown under 3D organotypic cell culture conditions. Studies are ongoing to confirm the role of Axl in the initiation and progression of melanomas harboring NRAS mutations and determine whether Axl constitutes a therapeutic target for this melanoma sub-group.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5111. doi:10.1158/1538-7445.AM2011-5111
Abstract 5121: A targeted approach to identify activators of class IA phosphoinositide-3-kinase in cancers using tandem mass spectrometry
Authors: Asara John M. , Cantley Lewis C. , Engelman Jeffrey A.
Published: June 2012

A majority of human cancers exhibit aberrant activation of the phosphoinositide 3-Kinase (PI3K) pathway. In many cases this activation is due to mutations or amplifications in upstream activators of PI3K resulting in activation by direct binding of phosphotyrosine proteins (e.g., receptor tyrosine kinases (RTK) or adaptors such as IRS-1/2 or Gab1/2) to the p85 regulatory subunit of PI3K. To date, experiments to identify the mechanisms of PI3K activation in cancer have remained difficult and cumbersome. Since inhibitors of many of these RTKs are either already approved or in clinical trials, identification of the major upstream activators of PI3K in a given tumor could suggest the appropriate therapeutic intervention. Here we present a quantitative mass spectrometry approach (targeted ion MS/MS – TIMM) for identifying the upstream activators of PI3K in cancer cell lines grown on plastic as well as in tumor Xenografts. Importantly, we show that in a series of non small cell lung carcinoma cell lines, it is possible to predict the appropriate drug for treating the cells based on the upstream activator of PI3K as revealed by TIMM. Specifically, we identified how PI3K is activated in MET amplified, EGFR and EML4-ALK driven non-small cell lung cancers (NSCLCs) both in vitro and in vivo, and were successful in identifying the optimal RTK inhibitor treatment for the H1703 NSCLC cell line. Additionally, this technology revealed that TORC1 inhibition leads to activation of PI3K via IRS-2, not IRS-1 in KRAS mutant cell lines.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5121. doi:10.1158/1538-7445.AM2011-5121
Abstract 5126: Development of a high throughput quantitative phosphoproteomic method to study trastuzumab resistance
Authors: Boyer Alaina P. , Alvarez Sophie , Boyne Michael , Hicks Leslie , Bose Ron
Published: June 2012

Her2 is a receptor tyrosine kinase that is overexpressed in approximately 20-30% of human breast cancers and affects clinical prognosis and outcome of these cancers. Treatment of Her2-positive breast cancer with the monoclonal antibody, trastuzumab, has improved the outcomes and prognosis for patients with this breast cancer subtype. However, development of resistance to trastuzumab is a major clinical problem in metastatic Her2-positive breast cancer. Many of the known mechanisms of trastuzumab resistance cause changes in protein phosphorylation patterns and therefore we have employed quantitative phosphoproteomics to further study trastuzumab resistance. Quantitative phosphoproteomics is a powerful method to comparatively identify phosphorylated proteins between different samples. Using electrospray tandem-mass spectrometry coupled with immunoaffinity purification and titanium dioxide (TiO2) phosphoenrichment, we wanted to identify differentially phosphorylated proteins in a pair of trastuzumab sensitive and resistant cells. We used stable isotope labeling of amino acids in cell culture (SILAC) to obtain quantitative information on the proteins identified by mass spectrometry. We have obtained quantitative data on two trastuzumab sensitive and resistant pairs of Her2 overexpressing human breast cancer cell lines (BT474 and SKBR3). 275 proteins were identified in the phosphoenriched fraction from the SKBR3 pair with 83 proteins reporting a ratio. 769 proteins were identified in the phosphoenriched fraction from the BT474 pair with 683 proteins reporting a ratio. Treatment of the trastuzumab resistant cells with Her2 siRNA drastically resulted in cell death indicating the sustained dependence of these cells on Her2 despite the resistance to Her2 targeted therapy. RNAi experiments, in combination with trastuzumab exposure, of candidate proteins expressing an increase in phosphorylation will be tested further to determine if trastuzumab resistance can be reversed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5126. doi:10.1158/1538-7445.AM2011-5126