bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3704: Gemtuzumab ozogamicin conjugated gold nanoparticles induce cell death in D1.1 leukemia cells when exposed to a radiofrequency field
Authors: Willborn Katherine A. , Glazer Evan S. , Briggs Katrina L. , Curley Steven A.
Published: February 2011

Introduction: Acute T-cell leukemia cells may express the marker CD33. Gemtuzumab is a monoclonal antibody against CD33, used clinically in combination with ozogamicin, a cytotoxic antibiotic, for treatment of acute leukemias. It has been previously demonstrated that gold nanoparticles delivered intracellularly and exposed to nonionizing radiofrequency (RF) fields induce hyperthermic cytotoxicity. RF fields are not cytotoxic without the presence of nanoparticles. The purpose of this experiment is to investigate cell death in cells treated with gemtuzumab ozogamicin conjugated to gold nanoparticles after RF field exposure.

Methods: D1.1 (acute T-cell leukemia) cells were plated in 60mm cell culture dishes. Two groups (n=3 in each group) of plated cells were treated for two hours with gemtuzumab ozogamicin conjugated to 10nm solid gold nanoparticles in media. One group of cells was then exposed to a radiofrequency field for a total generator energy of 1500 W-minutes. The control cells were treated with the same gold conjugate, but were not exposed to an RF field. Two days later, cell viability, apoptosis, and death was determined by flow cytometry. The groups were compared with Student's t-test.

Results: RF fields induced apoptosis in 58.7% ± 3.95% of treated cells, compared with 8.66% ± 0.56% of the controls (p=0.002). The viability after RF exposure was 13.9% ± 6.11% while gemtuzumab ozogamicin gold conjugate alone decreased viability to 74.2% ± 0.85% (p=0.003). The temperature of the media was kept below 40°C to avoid nonspecific cell injury.

Conclusions: Multimodality therapy (chemotherapy and nanoparticle-mediated hyperthermia) induced significantly more death in acute T-cell leukemia cells than antibody-chemotherapy alone. Antibodies specific to molecular targets on cells permit targeting of the heat mediator (nanoparticles) while preventing non-specific delivery of both nanoparticles and chemotherapeutic agent. The use of non-invasive RF fields to specifically induce cell death by hyperthermia with nanoparticles may present new options for the treatment of leukemias.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3704.
Abstract 3710: Facile Synthesis and Characterization of Pyropheophorbide-a -Taxol Conjugate
Authors: Kim Hyo Jun , Gantumur Battogtokh , Kim Sook Hee , Bae Sumi , Ahn Woong Shick
Published: February 2011

Chlorin-based photosensitizers are considered the ideal photosensitizer (second generation) in photodynamic therapy (PDT). They have improved efficacy and have lesser side effects compared to porphyrin-based photosensitizers (first generation). Paclitaxel (PTX) is one of the most effective and widely used anticancer a drug. Although chemotherapy is a powerful and important modality in cancer treatment, the effectiveness of the chemotherapy is limited due to systemic toxicity and side effects.

For the improvement of photosensitizers (chlorin; apoptosis, necrosis, local immune response) and increased anticancer drug (PTX; apoptosis), we combined photosensitizer and PTX.

For the synthesis, naturally occurring chlorin derivative was used as a starting material. It was prepared with the acidic treatment of methyl pyropheophorbide-a from Spirulina maxima algae according to the previously reported methods [1-2]. PTX should contain three hydroxyl groups at 21-position and 1-position and 7-position, respectively. A hydroxyl group at 21-position has more reactivity than others. To the synthesize target compound, we carried out an esterification reaction between the hydroxyl group at 21-position for the PTX moiety and carboxyl group at 173-position for the pyropheophorbide-a. The conjugate was performed as reaction the pyropheophorbide-a and the commercial PTX by using DMAP/DCC coupling agent.

The structure of chlorin- based PTX conjugate was fully characterized by 1D and 2D NMR techniques and also by MALDI-MS and UV-Vis. As well as, the conjugate was determined the purity of 99.45% by analytical reversed phase HPLC at 660 nm.

This conjugate can be applied as anticancer agent and photosensitizer for PDT. This compound is promising to be able to afford some synergistic effects in tumor cell.

References:

1. Smith, K. M.; Goff, D. A.; Simpson, D. J.; J. Am. Chem. Soc. 1985, 107, 4946-4954.

2. Galindev, O.; Badraa, N.; Dalantai, M.; Sengee, G.; Dorjnamjin, D.; Shim, Y. K. Photochem. Photobiol. Sci. 2008, 7, 1273-1281.

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3710.
Abstract 3722: Ovarian suppression with gonadotropin-releasing hormone analogues (Gn-RH) in high risk premenopausal breast cancer patients (PM-HR-BC): decrease of T regulatory cells (T-regs) and of vascular endothelial growth factor (VEGF)
Authors: Recchia Francesco , Candeloro Giampiero , Necozione Stefano , Desideri Giovambattista , D'Andrea Maurizio S. , Recchia Cornelia O.C. , Rea Silvio
Published: February 2011

Background: In a previous phase II study we have shown an improvement of disease-free survival and overall survival of 180 premenopauseal high-risk early breast cancer patients (PM-HR-BC) treated with ovarian suppression with Gn-RH and a chemotherapy tailored to the peculiar clinical situation. Aim of this study was the evaluation of VEGF and T-regs in a group of PM-HR-BC patients treated with the above mentioned therapy.

Methods: From 01/2006 to 12/2008 54 patients with PM-HR-BC were entered into this study. At baseline, one week before chemotherapy, patients received a Gn-RH analogue which was continued for five years. Tailored adjuvant chemotherapy, included anthracycline and taxane based regimens. 11 patients with triple negative tumors, received a platinum-based consolidation high-dose chemotherapy with (n. 7) or without (n. 4) autologous peripheral blood progenitor cell transplantation, respectively. Radiation therapy was administered to all patients that had received breast conserving surgery. Patients received a strong psychological support, biphosphonates, and after the end of chemotherapy, estrogen receptor positive (ER+) patients received an aromatase inhibitor.

Results: Characteristics of patients. Median age was 41 years (range, 26-44 yrs). 79% of patients were ER+, while 21% were estrogen receptor negative. Total number of positive nodes was 145, with a mean of 2,7 for each patient (range 0-25). G3 tumors were observed in 33% of patients, C-erB-2 status (FISH) was positive in 17% of patients, while mean Ki-67 value was 27%. Serum estradiol was suppressed to values < 40 pg/mL in all patients. After six months and one year, a statistically significant decrease was observed in T-regs number (p< 0.005) and VEGF (p< 0.005) with respect to baseline values. No impairement of cognitive function was observed with respect to baseline conditions. After a median follow-up of 26 months, 4 years progression-free survival and overall survival rate were 97% and 100%, respectively. Conclusions: These data show that ovarian suppression with a Gn-RH analogue decreases T-regs and VEGF, and seems to improve the expected outcome of PM-HR-BC patients through immunologic mechanisms.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3722.
Abstract 3725: Bortezomib-induced neuropathic pain: Evaluation of anti nociceptive effect of a new analgesic compound
Authors: Canta Annalisa , Meregalli Cristina , Carozzi Valentina Alda , Oggioni Norberto , Chiorazzi Alessia , Tredici Giovanni , Lanza Marco , Letari Ornella , Caselli Gianfranco
Published: February 2011

Bortezomib (Btz) is a highly effective and widely used antineoplastic drug for the treatment of many tumors, primarily multiple myeloma. Despite its effectiveness, the use of this proteasome inhibitor is limited by its toxicity. Among Btz side effects, peripheral neurotoxicity is a major and clinically relevant problem. In fact, since the earliest Phase I and Phase II studies, sensory peripheral neuropathy emerged as a dose-limiting side effect. Moreover, a significant proportion of Btz-treated patients developed severe neuropathic pain. The treatment of Btz-induced neuropathic pain is extremely difficult with the currently available drugs and still remains a major challenge for oncologists and neurologists. Aim of this study was to demonstrate if the new analgesic compound CR4056 could reverse the neuropathic pain induced by a chronic treatment with Btz in the rat. The anti-nociceptive and dose-dependent effect of CR4056 was assessed in a well-established experimental rat model in which Btz was administered to Wistar rats (0.20 mg/kg three times/week for 8 weeks) followed by treatment with CR4056 (6, 20 or 60 mg/kg daily p.o.) or buprenorphine administered 28,8 µg/kg/day p.o. for 2 weeks. Body weight change, hematological and histopathological parameters, nerve conduction velocity (NCV) and sensory behavioral tests were evaluated during the experiment. No remarkable effect of Btz administration or of any dose of CR4056 was observed in the hematological parameters and blood chemistry. After 8 weeks of treatment, Btz induced a severe reduction in NCV; this impairment was not reversed by CR4056 or by buprenorphine. The behavioral assessment showed a significant mechanical allodynia in rats treated with Btz for 8 weeks (delta vs. controls −6 g). Buprenorphine significantly reduced Btz-induced allodynia during the fist day by about 63%. However, this effect was lost during the following days of treatment (i.e 20% reduction on the 3rd day). The mid and high dose of CR4056 (20 and 60 mg/kg) showed a significant (i.e > 80 %) reduction of allodynia till the 3rd day of administration, afterwards a slight reduction in efficacy was recorded. On the contrary, the oral daily administration of 6 mg/kg CR4056 persistently counteracted Btz-induced allodynia, showing an even complete reversion after 2 weeks of treatment. The present study gives evidence for a relevant and persistent anti-allodynic effect of CR4056, suggesting a role for CR4056 in the management of neuropathic pain in patients treated with Btz.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3725.
Abstract 3729: Prognostication using Hif-1 alpha may require simultaneous nicotine assessment
Authors: Warren Graham W. , Singh Anurag , Randall Marcus , McGarry Ronald , Kudrimoti Mahesh , Rangnekar Vivek
Published: February 2011

Background: Current cancer diagnosis and treatment increasingly incorporates biomarker expression in the prognostication and therapeutic stratification of cancer patients; however, there is no accurate assessment of exposure to environmental agents such as nicotine that may significantly impact biomarker expression patterns.

Methods: Athymic nude mice with human H460 lung cancer cell xenografts were stratified into three treatment groups (n=5 per group) beginning when tumors reached 5 mm in maximal dimension: control (CON), short term nicotine (STN, 60 ug subcutaneously every other day x 6 days), or long term nicotine (LTN, 60 ug SC every other day until tumor explant). All tumors were explanted at endpoint defined when tumors reached 15 mm in maximal dimension. Tumors were analyzed immunohistochemically for hypoxia (using CA-IX expression) and hypoxia-inducible factor 1-alpha (Hif-1) expression. This protocol was approved by the Institutional Animal Care and Use Committee at the University of Kentucky.

Results: Nicotine administration had no effect on tumor growth rate and all tumors reached endpoint within a median of 9 days following treatment initiation (range 8-11 days). Areas of hypoxia were defined using CA-IX expression. Tumor hypoxia was not significantly affected by nicotine administration. Moreover, Hif-1 expression was only minimally increased in areas of hypoxia in CON animals. In STN animals, Hif-1 expression was very similar to CON animals. In contrast, LTN animals demonstrated widespread marked increases in Hif-1 expression at the viable: necrotic tumor interface. Separately, xenografts removed at day 6 demonstrated similar widespread Hif-1 expression confirming that Hif-1 expression is significantly increased after 6 days of nicotine administration.

Conclusions: This study demonstrates that widespread Hif-1 expression may be dominated by the presence of nicotine rather than hypoxia. Furthermore, acute removal of nicotine results in the restoration of basal low-level Hif-1 expression within 3 days of nicotine removal. This data may significantly limit the prognostic utility of clinical Hif-1 expression in the absence of simultaneous nicotine assessment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3729.
Abstract 3734: TLE3 expression is predictive of response to chemotherapy in NSCLC
Authors: Ross Douglas T. , Seitz Robert S. , Ring Brian Z. , Wang Yanling , Beck Rodney A. , Soltermann Alex
Published: February 2011

Purpose: Previously we have shown that the wnt pathway marker TLE3 is associated with outcome in taxane-treated breast cancer but not in patients treated with anthracycline-only supporting TLE3 as a taxane predictive marker. The purpose of this study was to assess the association between TLE3 expression and outcome in NSCLC patients treated with chemotherapy as opposed to those treated with a surgical resection and or radiation alone.

Experimental Design: We performed TLE3 immunohistochemistry staining on NSCLC cancer specimens from the University of Zurich, Switzerland. Most advanced stage chemotherapy treated patients at this institute receive a taxane containing regimen. Local and distant recurrences within the first five years of follow-up were analyzed using Kaplan Meier, Cox proportional hazard, and multivariate analysis to assess an interaction between TLE3 expression and response to chemotherapy.

Results: TLE3 staining with a monoclonal antibody was associated with outcome only when patients received chemotherapy (HR=0.42 p=0.0047 n=68) while no association was identified in untreated patients (p=0.83 n=300). Multivariate analysis showed that TLE3 was independent of stage, grade and age. An interaction test between TLE3 expression and chemotherapy response was significant (p=0.036).

Conclusion: High TLE3 expression is associated with response to chemotherapy in NSCLC. This is consistent with data in breast cancer where TLE3 is associated with response to taxane therapy. Further study in randomized cohorts is warranted to confirm TLE3 as a predictive marker for response the taxane therapy in NSCLC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3734.
Abstract 3739: Clinicopathologic significance of ERCC1, Thymidylate Synthase and Glutathione S-Transferase P1 expression for advanced gastric cancer patients receiving adjuvant 5-FU and cisplatin chemotherapy
Authors: OH SUNG YONG , Kim Ki Han , Kim Sung Hyun , Lee Suee , Kwon Kyung-A , Jang Jin Seok , Kim Min Chan , Kim Su-Jin , Kim Hyo-Jin , Kwon Hyuk-Chan
Published: February 2011

Background: The aim of this study was to determine whether expressions of the excision cross-complementing (ERCC1), thymidylate synthase (TS) and glutathione S-transferase P1 (GSTP1) predict clinical outcome in patients with advanced gastric cancer treated with adjuvant 5-fluorouracil (5-FU) and cisplatin (FP) chemotherapy.

Patients and methods: The study population consisted of 149 advanced gastric cancer patients. After curative resection, if the patients had stage II or higher disease, we administered adjuvant FP combination chemotherapy. The FP regimen was as follows. Cisplatin 60 mg/m2 was administered intravenously on day 1, and 5-FU 1,000 mg/m2 was administered intravenously on days 1 to 5. This regimen was repeated every three weeks. The expressions of ERCC1, TS and GSTP1 of primary tumors were examined by immunohistochemistry. Clinicopathologic information, including tumor size, histological grade, Lauren's classification, Borrmann type, vascular invasion, ratio of involved lymph nodes to dissected nodes, stage, carcinoembryonic antigen (CEA), and disease-free survival (DFS) and overall survival (OS) were evaluated with respect to the expressions of ERCC1, TS and GSTP1.

Results: Median follow-up duration was 65.2 months, and median patient age was 51 ± 11 years (range 24-75). The positive rates of ERCC1, TS, and GSTP1 were 63.1%, 76.5%, and 40.3%, respectively. ERCC1 and GSTP1 expression was significantly correlated with size (p = 0.040, p = 0.018, respectively). Positive relationships were found between the expression of TS and Borrmann type (p = 0.030) and stage (p = 0.041). Univariate analysis showed that Lauren classification, stage, positive lymph node ratio, and type of operation were associated with both DFS and OS. ERCC1 and GTP1 were significant variables affecting OS (p = 0.005, p = 0.024, respectively). Patients who were both ERCC1 and GSTP1 positive had longer survival (p = 0.003). Multivariate analysis revealed that stage and positive lymph node ratio were independently associated with DFS and OS. Expression of both ERCC1 and GSTP1significantly impacted on OS (hazard ratio = 0.069, p = 0.021).

Conclusion: Immunohistochemical studies for ERCC1 and GSTP1 were useful in prediction of the clinical outcome in advanced gastric cancer patients treated with adjuvant FP.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3739.
Abstract 3742: FOXC1 is a potential prognostic marker with functional significance in basal-like breast cancer
Authors: Wang Jinhua , Ray Partha , Qu Ying , Sim Myung-Shin , Shamonki Jaime , Bagaria Sanjay , Ye Xing , Liu Bingya , Walter Michael , Hoon Dave , Giuliano Armando , Cui Xiaojiang
Published: February 2011

Although basal-like breast cancer (BLBC) is a distinct molecular subtype, it has no defining gene or protein marker that can be used as the basis for therapy. Estrogen receptor (ER) and HER2 receptor guide treatment of luminal and HER2 breast cancer subtypes, respectively, but chemotherapy is still the only systemic treatment available for BLBC. In view of its poor clinical outcomes, high proliferation rates and preferential metastasis to the brain, there is an urgent need for effective targeted therapy of BLBC.

In a previous study using cDNA microarray analyses and immunohistochemistry of archival breast cancer tissue specimens, we showed consistent and exclusive overexpression of FOXC1 in BLBC versus other molecular subtypes. FOXC1 overexpression was significantly associated with poor overall survival and brain metastasis. In the present study, we show that overexpression of FOXC1 in breast cancer cells with low endogenous FOXC1 levels increased cellular proliferation, migration, and invasion, and induced epithelial-mesenchymal transition. Knockdown of FOXC1 by shRNA in breast cancer cells with high endogenous FOXC1 expression had the opposite effect. Cell signaling studies showed that FOXC1 activated the NF-κB pathway by upregulating the peptidyl-prolyl isomerase Pin1 and downregulating the ubiquitin ligase SOCS-1. Pin1 binds to p65, inhibits the p65 interaction with IκBα, and thus enhances p65 nuclear localization and protein stability. SOCS-1 directly interacts with p65 and promotes its degradation. FOXC1 knockdown reduced p65 protein levels and NF-κB activity. The importance of the NF-κB pathway for FOXC1 function was confirmed by the increased sensitivity to NF-κB inhibitors in cells that overexpressed FOXC1. Because BLBC under-expresses ER, FOXC1 was inversely associated with ER, as expected. Interestingly, FOXC1 repressed ER transcription and activity via NF-κB signaling, and FOXC1 overexpression switched MCF-7 breast cancer cell growth from estrogen-dependent to estrogen-independent. This may explain why ER is not normally detected in BLBC.

These results suggest that FOXC1 may be a specific diagnostic and prognostic biomarker for BLBC and may play an important role in regulating aggressive traits associated with BLBC. It might also serve as a potential molecular therapeutic target for BLBC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3742.
Abstract 375: Targeting pericytes in ovarian carcinoma
Authors: Lu Chunhua , Shahzad Mian M.K , Moreno-Smith Myrthala , Lin Yvonne G. , Jennings Nicholas B. , Allen Julie K. , Hu Wei , stone Rebecca L. , Matsuo Koji , Landen Charles N. , Coleman Robert L. , Sood Anil K.
Published: February 2011

Purpose: Pericytes interact with and closely regulate endothelial cell function. Platelet-derived growth factor (PDGF)-BB/PDGF receptor (PDGFR) β plays a critical role in pericyte regulation, therefore highly specific inhibitors of this target are needed. We hypothesize that dual targeting of endothelial cells and pericytes provide a more efficacious antiangiogenic approach for ovarian cancer therapy. Experimental Design: The therapeutic efficacy of targeting endothelial cells (bevacizumab) and/or pericytes (PDGF-aptamer, AX102) was examined using SKOV3ip1 and HeyA8 orthotopic ovarian cancer models. The biomarkers, such as microvessel density (MVD), vascular maturation (pericyte coverage), and proliferative index (proliferating cell nuclear antigen, PCNA) were examined. Results: Bevacizumab inhibited tumor growth by 48% and 45% in the SKOV3ip1 and HeyA8 models, respectively. AX102 had minimal effect on the HeyA8 model, but increased tumor growth in the SKOV3ip1 model. However, bevacizumab plus AX102 was more effective than bevacizumab alone, and reduced 76 - 88% of tumor growth in both models. A longitudinal study using bioluminescence imaging in the HeyA8 model showed that a combination of bevacizumab, AX102 and paclitaxel caused tumor reduction by 65% (based on bioluminescence imaging). MVD and proliferative index counts were significantly reduced in the bevacizumab treatment groups, and pericyte coverage was significantly decreased in the AX102 treatment groups. Conclusions: Dual targeting of endothelial cells and pericytes demonstrates therapeutic potential as a unique anti-vascular approach in treating ovarian carcinoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 375.
Abstract 3751: Clinical correlation between Cyclin D1, pAKT, and FOXO1A in African American and Latino Breast Cancer Patients
Authors: Lee Jane , Wu Yanyuan , Sarkissyan Marianna , Elshimali Yayha , Vadgama Jaydutt V.
Published: February 2011

Breast cancer is the most common cause of cancer amongst women and is the seventh highest cause of death in this population in the United States. Statistics indicate that minority and underserved populations have worse overall survival from breast cancer. Clinical data from our laboratory have shown that African American and Latino breast cancer patients with HER2/neu overexpression and high pAKT have poor outcomes. In vitro data on breast cancer cells have shown that Cyclin D1 is up-regulated by the PI3K/AKT pathway, while the FOXO1A transcription factor is down-regulated. Hence, the purpose of our current study was to evaluate clinical correlation between pAKT, Cyclin D1, and FOXO1A in tissues from the same patient. We used the same cohort of patients from our previous study. Immunohistochemistry was performed on paraffin tissues from African American and Latino women with breast cancer. Results indicate that patients with HER2/neu overexpression were likely to be Cyclin D1 positive. In addition, those with high pAKT were also Cyclin D1 positive, thus indicating that HER2/neu overexpression might be associated with increase in Cyclin D1 and pAKT. Interestingly, breast tumors with high pAKT and low FOXO1A were also Cyclin D1 positive. In summary, immunohistochemical analysis and subsequent correlation of biomarkers associated with the HER2-PI3/AKT pathway, that include pAKT, Cyclin D1 and FOXO1 may provide better assessment of clinical outcome of breast cancer patients and therefore better target therapies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3751.
Abstract 3752: Folate receptor expression prevalence across multiple tumor types and concordance between matched primary and metastatic tumor pairs
Authors: Li Hewei , Wu Qiuyan , Xing Guan , Teslenko Vic , Tornout Jan Van , Ahlers Christoph M. , Huang Fei
Published: February 2011

Background: The human folate receptor (FR) is selectively overexpressed in a number of human epithelial malignancies, including ovarian, renal, and prostate cancers. Folate-conjugated therapies provide a strategy for targeting patients with FR expressing tumors. Identification of patients with FR expressing tumors is important to increase the opportunity for clinical benefit from folate conjugated therapies. The goals of this study are to determine the prevalence of FRα expression in various solid tumors, and to determine the concordance of FRα expression between primary and matched metastatic tumors. This concordance could help to determine if primary tumor samples would be suitable for assessing FR expression in metastatic tumors. Little information on this is currently available.

Methods: FRα expression was analyzed by immunohistochemistry (IHC) in 339 primary tumors covering a wide range of human tumor types. Concordance of FRα status in 86 archived primary and matched metastatic breast, renal, endometrial and ovarian tumors was assessed by IHC.

Results: FRα expression prevalence was moderate to high in primary tumors of lung (13/29; 45%), renal (66/115; 57%), colon (17/27; 63%), prostate (8/12; 67%) and ovarian (22/29; 76%). FRα expression frequency was found relatively low in breast tumor (26/127; 20%) which is consistent with previous reports. Furthermore, for the first time, we showed a good concordance (81.4%) of FRα expression between paired primary and metastatic ovarian, endometrial, renal and breast malignancies.

Conclusions: FRα is frequently overexpressed in ovarian, prostate, colon and renal tumors, suggesting these tumor patients might benefit more from folate conjugated therapies. New findings from this study show that FRα expression is maintained in metastatic tumors, indicating the utility of archived primary tumor samples for determining the FR positivity of metastatic tumors in patients receiving a folate conjugated agent.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3752.
Abstract 3755: Phosphorylated STAT3 in polymorphonucleur cells (PMNs) as a potential surrogate marker to evaluate the efficiency of sorafenib therapy
Authors: Wu Jun , Wang Yan , Yen Yun , Reckamp Karen L.
Published: February 2011

Sorafenib tosylate (BAY43-9006) is an oral multi-kinase inhibitor which targets c-Raf and b-Raf, and thus interrupts this critical oncogenic pathway. Sorafenib has been successfully applied in the clinic to treat patients with certain types of renal cell carcinoma and hepatocellular carcinoma. Signal transducer and activator of transcription 3 (STAT3) plays an important role in the pathogenesis and progression of cancer. Furthermore, the inhibition of STAT3 by sorafenib has been previously described. In order to evaluate STAT3 within polymorphonucleur cells (PMNs) as a potential surrogate marker of phosphorylated STAT3 (p-STAT3) in tumor tissue, we proposed a preclinical animal model to examine the relationship of p-STAT3 as in tumors and PMNs during treatment with sorafenib.

Both normal balb/c mice (female and male, 10 weeks old) and immunodeficient NOD/SCID/IL2Rgamma null (NSG) mice (female at 10 weeks old) were used in the animal studies. Human ovarian cancer cells OV-90 were inoculated subcutaneously into the dorsal area of NSG mice to create a xenograft model. Sorafenib was given at 50 or 100 mg/kg in balb/c mice and 60 mg/kg in NSG mice via oral gavage for three doses before tissue sampling. Two hours after the last drug treatment, the mice were euthanized and the blood and tumor tissues were harvested for pSTAT3 level assays. Phosphorylated STAT3 expression levels were analyzed by flow cytometry and western blot.

Suppression of pSTAT3 levels in PMNs from balb/c mice was observed in 1/3 of mice at 50 mg/kg and 2/3 of mice at 100 mg/kg by both western blot and flow cytometry, however the difference between the vehicle control and sorafenib group was not significant. Suppression of pSTAT3 levels in OV-90 tumor tissue from sorafenib-treated NSG mice was significant by western blot analysis. However the suppression of pSTAT3 on the PMNs from the corresponding NSG mice was minor by flow cytometry analysis. In conclusion, in this murine model, pSTAT3 in PMNs was not a consistent surrogate marker for the evaluation of sorafenib activity. Based on the emerging data linking escalating doses of sorafenib to STAT3 inhibition in cancer cells, and the tumor responses seen with increasing doses, evaluation of surrogate biomarkers to predict clinical efficacy will be useful.

Supported by NCI K12 CA 01727 and the Phase One Foundation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3755.
Abstract 3758: Global expression profiling of highly degraded RNA by DASL assay from formalin-fixed paraffin-embedded lung tissue
Authors: Su Li
Published: February 2011

Microarray-based genome-wide gene expression analysis on fresh frozen tissue is widely used in cancer studies. Compared to fresh frozen tissues, formalin-fixed paraffin-embedded (FFPE) tissues are more accessible in clinical setting for genomic analysis, but its use has been hampered by technical difficulty. Recently, cDNA-medi¬ated annealing, selection, extension, and ligation (DASL) assay was developed by Illumina (San Diego, CA) to investigate the global gene expression profiling of highly degraded RNA from FFPE samples. In this study, we tested the potential of the Whole-Genome DASL Assay for Expression Profiling in FFPE Samples (covering up to 24,000 annotated probes) in biomarker discovery for lung disease research. We obtained biopsy tissue from two lung cancer patients and six acute respiratory distress syndrome (ARDS) patients. Five 5 μm-thick slides from each specimen were evaluated by a pathologist for tumor cellularity (lung cancer) or diffuse alveolar damage (ARDS), and RNA was extracted using commercially available (Roche High Pure RNA Paraffin) kits. The DASL expression profiles on 2 ARDS FFPE-extracted RNA samples had poor quality as determined by low detection rate (<25%) and low array-wide average signals. The other 6 (75%) of the RNA samples had a median detection of 8,442 genes with a range between 7,306 and 10,616 genes. These array data also had strong array-wide average signals and low noise. Our study demonstrated that DASL expression profiling technology, genome-wide expression profiles can be successfully generated from FFPE lung tissues.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3758.
Abstract 3761: Tissue processing time affects the expression of phosphoproteins after breast cancer resection
Authors: Gao Ming , Huang Ta-Chen , Chen Chi-Long , Tu Hui-Ching , Feng Wen-Chi , Lu Yen-Shen , Yeh Pei-Yen , Kuo Wun-Hon , Chang King-Jeng , Hsu Chih-Hung , Cheng Ann-Lii
Published: February 2011

Background: Many phosphoproteins are among the targeted pathways of cancer signal transduction and are therefore candidates for cancer biomarkers. However, the collection procedures of biospecimens might affect the expression of the phosphoproteins significantly because the time interval between surgical removal of tissues and fixation, i.e. warm ischemia time, are variable. This study aimed to evaluate the stability of phosphoprotein expression under the differential tissue processing time.

Materials and Methods: Specimens were obtained from patients who received mastectomy at National Taiwan University Hospital. Each specimen was cut into 5 pieces, stored at 4oC for 10 minutes, 2, 4, 6, and 24 hours before further processing. At each assigned time point, a small section of the specimen was fixed immediately in 10% buffered formalin, then paraffin embedded for immunohistochemical stains (IHC) of phospho-Akt (p-Akt) and phospho-Erk (p-Erk). The remaining specimens were snap frozen in liquid nitrogen, followed by homogenization in RIPA buffer. Western blot analysis was performed for phospho-Akt (p-Akt), total Akt, phospho-Erk (p-Erk), total Erk, phospho-STAT3 (p-STAT3), and total STAT3 staining.

Results: Western blot analysis showed that the expression of p-Erk declined significantly as early as 2 hours. The expression of p-Akt was relatively stable and decreased slightly after 24 hours. The expression of p-STAT3 gave variable results. The expression of total Akt, Erk, and STAT3 remain unchanged at each assigned time point. The immunohistochemical expression of p-ERK declined in a time-dependent manner whereas no obvious p-Akt decrease was detected.

Conclusions: Our results indicate that the expression of some phosphoproteins by IHC may be affected by warm ischemia time. Caution should be given to the interpretation of the results of these biomarkers.

(The study was supported by the grant NSC 98-3112-B-002-038)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3761.
Abstract 3768: Comparison of breast density measured by dual energy x-ray Absorptiometry (DXA) with mammographic density among adult women
Authors: Maskarinec Gertraud , Morimoto Yukiko , Daida Yihe , Laidevant Aurelie , Shepherd John A. , Novotny Rachel
Published: February 2011

Mammographic density, the distribution of fat, connective, and epithelial tissues in the female breast, is strongly associated with breast cancer and has been used as a biomarker for breast cancer risk among adult women. While use of mammography is limited due to concerns about radiation exposure, Dual Energy X-ray Absorptiometry (DXA), commonly available in medical care settings, has very low radiation. In this analysis, we compared breast density measured by DXA with mammographic density among adult women. We recruited 101 participants aged 30 years and older with a normal mammogram (defined as BIRADS categories 1 through 3) through Kaiser Permanente Hawaii. After completing a questionnaire asking for demographic, reproductive, anthropometric, and medical information, all women received DXA scans of both breasts, as well as of the whole body, using a GE Lunar Prodigy Bone Densitometer. On the DXA scans, the breast image was manually outlined and the following measures were computed: percent fibroglandular volume density (% FGV), absolute fibroglandular volume, total breast area, and volume. After digitizing the most recent craniocaudal (CC) and mediolateral (ML) mammograms, total breast area, dense area, and percent density were estimated using a computer-assisted method (Cumulus). For the overall population, mean % FGV was 38.8±14.3%, and mean percent density was 31.9±18.2% for CC views and 28.3±16.2% for ML views. Both DXA and mammographic measures showed high correlations between left and right breasts ranging from 0.85 to 0.98 (p <0.0001). When comparing DXA and mammographic measures, the correlation coefficients between % FGV and percent density were 0.76 for both CC and ML views (p <0.0001). Associations with common risk factors showed similar patterns for DXA and mammographic densities; in particular, the inverse associations with BMI and age at menarche were evident for both methods. Multilinear regression with stepwise selection indicated an explained variance of 0.57 for % FGV alone and 0.58 for % FGV plus parity. The current comparison suggests that DXA has the potential to provide a low-radiation option to evaluate breast density for women who do not undergo mammography. This may allow improved individualized risk assessment to identify women for targeted prevention strategies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3768.
Abstract 3769: Use of combined levels of dietary intake from FFQ and plasma biomarkers to assess association between antioxidant exposures and prostate cancer risk
Authors: Kong So Yeon J. , Goodman Michael , Bostick Roberd M.
Published: February 2011

Epidemiologic studies evaluating the relation between specific nutrients and risk of disease typically rely either on questionnaire-based or on biomarker-based measures of exposure; however both of these approaches may be affected by systematic error. In this study, we aim to improve assessment of antioxidant exposure through a combined measure reflecting both plasma measurements and food frequency questionnaire (FFQ)-derived levels of intake. This combined measure of exposure is examined in relation to risk of prostate cancer. In this community-based case-control study, dietary intake was assessed through the validated Block FFQ that allowed estimating daily intakes of various antioxidants. Archived plasma samples were analyzed using high performance liquid chromatography to determine circulating levels of carotenoids (lycopene, α-carotene, β-carotene, lutine, and β-cryptoxanthin), retinol (vitamin A), and α-tocopherol (vitamin E). Cases and controls were categorized as having “high” or “low” for each antioxidant's dietary intake and plasma level based on the median value among controls. Each participant was then included into one of the following four categories: (1) low dietary intake and low plasma level; (2) low dietary intake and high plasma level; (3) high dietary intake and low plasma level; and (4) high dietary intake and high plasma level. The combined measurements of high dietary intake and high plasma levels of lycopene, β-cryptoxanthin, and α-tocopherol demonstrated a statistically significant association with prostate cancer risk with adjusted odds ratios of 0.46 (0.23-0.91), 0.43 (0.22-0.84), and 0.32 (0.14-0.72), respectively. The combined measurements of high dietary intake and low plasma levels of β-carotene also demonstrated a statistically significant association (aOR = 0.43; 95% CI=0.20-0.91) with prostate cancer risk. These results tended to be stronger than the corresponding results for each measure of exposure considered individually. These findings provide evidence that the use of combined levels of dietary intake from FFQ and plasma biomarkers can improve the assessment of antioxidant exposure compared to use of either FFQ or plasma biomarkers separately.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3769.
Abstract 3770: Long-term variation in serum 25-hydroxyvitamin D concentration among participants in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial
Authors: Hofmann Jonathan N. , Yu Kai , Horst Ronald L. , Hayes Richard B. , Purdue Mark P.
Published: February 2011

Molecular epidemiologic studies of vitamin D and risk of cancer and other health outcomes usually involve a single measurement of the biomarker 25-hydroxyvitamin D [25(OH)D] in serum or plasma. However, the extent to which 25(OH)D concentration at a single time point is representative of an individual's long-term vitamin D status is unclear. To address this question, we evaluated within-person variability in 25(OH)D concentration across serum samples collected at three time points over a five year period among 29 participants in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial. Serial blood samples in the PLCO Cancer Screening Trial were obtained from each subject at the same time of year for each of the three collection events. This sample collection protocol provided a unique opportunity to evaluate within-person temporal variability beyond seasonal effects, which have been well-characterized in previous studies. The within-person coefficient of variation (CV) and intraclass correlation coefficient (ICC) were calculated using variance components estimated from random effects models. Spearman rank correlation coefficients were calculated to evaluate agreement between measurements at different collection times (baseline, +1 year, +5 years). The within-subject CV was 14.9% [95% confidence interval (CI) 12.4%-18.1%] and the ICC was 0.71 (95% CI 0.63-0.88). Spearman rank correlation coefficients comparing baseline to +1 year, +1 year to +5 years, baseline to +5 years, and the average of baseline and +1 year to +5 years were 0.65 (0.37-0.82), 0.61 (0.29-0.81), 0.53 (0.17-0.77), and 0.65 (0.34-0.83) respectively. After restricting to 23 non-Hispanic Caucasian subjects, the ICC was 0.75 (0.64-0.90) and slightly stronger pairwise correlations were observed: the correlation coefficients were 0.72 (0.42-0.87) for baseline vs. +1 year, 0.69 (0.37-0.87) for +1 year vs. +5 years, 0.58 (0.18-0.81) for baseline vs. +5 years, and 0.70 (0.37-0.87) for the average of baseline and +1 year vs. +5 years. These findings suggest that serum 25(OH)D concentration at a single time point may be a useful biomarker of long-term vitamin D status in population-based studies of various diseases. When possible, investigators might also consider taking multiple measurements from each subject at different times to reduce misclassification of vitamin D status, thereby improving the validity of risk estimates.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3770.
Abstract 3776: Third follow-up of a Dutch cohort occupationally exposed to chlorophenoxy herbicides, chlorophenols and dioxins
Authors: Boers Daisy , Portengen Lutzen , Bueno-de-Mesquita Bas , Heederik Dick , Vermeulen Roel
Published: February 2011

A retrospective cohort study of workers exposed to phenoxy herbicides, chlorophenols, and contaminants (2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and other polychlorinated dioxins and furans) has been conducted in two chemical factories in the Netherlands. TCDD levels at time of presumed last exposure were estimated based on back-extrapolation of predictive models derived from a cross-sectional survey on a sub-population of the full cohort. Analyses were done using Cox proportional hazard models, using attained age as the time-scale.

In analyses relating exposure to phenoxy herbicides to cancer mortality the previously reported increased risks for respiratory cancer, and non-Hodgkin's lymphoma in factory A could not be confirmed. However, increased risks were observed for all cancers in both factory A (HR=1.31; 95% CI 0.86-2.01) and factory B (HR=1.54; 95% CI 1.00-2.37). Increased risks for urinary cancers (HR=4.2; 95% CI 0.99-17.89) and genital cancers (HR=2.93; 95% CI 0.61-14.15) were observed in factory A, consistent with earlier reported results in this population. More detailed analyses showed that this increased risk for urinary and genital cancers in exposed workers was not due to selection of healthy controls and could not be attributed to specific products or departments.

The results of this study showed only slight increases in cancer mortality risk among phenoxy herbicide exposed workers. Analyses using estimated (back-extrapolated) TCDD are currently in progress. Preliminary analyses suggest an increased risk for non-Hodgkin lymphoma in TCDD exposed factory A compared to B.

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

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3776.