bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3456: Polychlorinated biphenyls and relative telomere length in the Anniston Community Health Survey: A cross-sectional study
Authors: Callahan Catherine , Pavuk Marian , Ren Xuefeng , Olson James R. , Bonner Matthew R.
Published: July 2016

Background: Polychlorinated biphenyls (PCBs) exposure is associated with increased risk of melanoma and non-Hodgkin lymphoma in some, but not all, epidemiologic studies. Although the carcinogenic mechanisms of specific PCB congeners remain to be fully explained, PCBs have been demonstrated to induce oxidative stress and inflammation and therefore may have deleterious impacts on telomere length. We investigated the association between serum concentrations of PCBs and telomere length in a highly exposed population.

Methods: The Anniston Community Health Survey (ACHS) is a cross-sectional study of 766 residents of Anniston, Alabama conducted between 2005 and 2007. We measured relative telomere length (RTL) with telomere/single copy gene ratio, in DNA extracted from peripheral blood samples in a subset of 57 African-American and 46 white participants, using monochrome multiplex quantitative polymerase chain reaction. We used multiple linear regression to estimate the association between 27 individual PCB congeners, and sum PCBs, adjusting for age, plate, and total lipids. All analyses were stratified by race. PCBs and RTL were log-transformed.

Results: ACHS participants had high serum concentrations of PCBs (mean sum PCBs among African-Americans = 14,687.69 ppt whole weight, SD = 23,515.46; mean sum PCBs white participants 6,088.32 ppt whole weight, SD = 8,750.57). RTL did not differ by race (mean RTL African-Americans = 0.94 SD = 0.08; mean RTL whites = 0.94, SD = 0.09). Among African-American participants, sum serum PCBs was not associated with RTL (0.70% per 100 ppt; 95% confidence interval = -9.43, 19.70). However, PCB 167 was weakly associated with higher RTL (2.35%, 95%CI = -8.56, 14.55), while PCB 157 was associated with shorter RTL (-4.36%; 95%CI = -15.63, 8.40). Among white participants, a 100 ppt increase in sum serum PCBs was associated with a 10.20% increase in RTL (95% CI = -7.10, 30.72). PCB 138, in particular, was associated with increased RTL among white participants (16.29%; 95% CI = 1.34, 33.46). Conversely, PCB 189 was associated with shorter RTL among white participants (-10.48%; 95%CI = -29.82, 14.19).

Conclusions: We did not observe consistent associations between serum concentrations of PCBs and RTL, suggesting that the association may vary by race and be congener specific. Our results are necessarily preliminary and are limited by the small number of participants. Additional measurements of RTL among ACHS participants are on-going.

Citation Format: Catherine Callahan, Marian Pavuk, Xuefeng Ren, James R. Olson, Matthew R. Bonner. Polychlorinated biphenyls and relative telomere length in the Anniston Community Health Survey: A cross-sectional study. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3456.
Abstract 3464: Low concordance with CEA tumor marker monitoring in colorectal cancer survivors
Authors: Palos Guadalupe , Gilmore Katherine R. , Chapman Patricia , Lewis-Patterson Paula , Bi Weiqi , Rodriguez Maria Alma
Published: July 2016

Purpose: To assess providers’ concordance with surveillance and risk reduction recommendations for colorectal cancer (CRC) survivors after completion of their curative treatment.

Patients and Methods:

This was a longitudinal study of survivors who met the following eligibility criteria: diagnosed with a primary colon or rectal cancer before their first visit (V) to the CRC Survivorship Clinic, adult survivor ≥ 18 years old, no evidence of disease, alive at the time of data abstraction, and 1-3 clinic visits between 9/01/2011 and 8/31/2014. Data were collected at V1 scheduled between 9/1/2011 and 8/31/2012. V2 was scheduled 9-15 months after the first visit. V3 was also scheduled 9-15 months after V2. Data sources were survivorship care plans, electronic medical records, and CRC survivorship algorithms. For an annual visit, CRC algorithms recommended history/physical exams (H & PEs), carcinoembryonic antigen (CEA) testing when previously elevated, and colonoscopies for surveillance of cancer recurrence. Concordance rates (CR) were measured as the percent of yes/no responses to whether the providers followed minimum standards for the 3 procedures. Demographic and clinical characteristics were also collected. Descriptive statistics were used to summarize all data.

Results:

81 of 117 CRC survivors who met all eligibility criteria were included in this sub-analysis. The number of survivors visits varied across time, V1 = 81, V2 = 56, and V3 = 36. Most survivors were male (51.9%) and Caucasian (66.7%). 67.9% reported being 5-8 years post-treatment. 58% were diagnosed with colon cancer and of those 61.7% were Stage IIIA-IV compared to 55.8% of rectal cancer survivors with advanced disease. Table 1 summarizes the percentage of CR rates across the 3 visits.

Conclusion:

CRs for H & PEs and colonoscopies remained high across the 3 visits. CRs were lowest for CEA recommendations. These low rates suggest further work is needed to determine barriers in clinical practice that limit use of CEA tumor marker monitoring in CRC survivors. Percent of Providers ConcordanceAlgorithm RecommendationVisit 1Visit 2Visit 3H & PE100100100CEA53.338.322.2Colonoscopy97.598.2100

Citation Format: Guadalupe Palos, Katherine R. Gilmore, Patricia Chapman, Paula Lewis-Patterson, Weiqi Bi, Maria Alma Rodriguez. Low concordance with CEA tumor marker monitoring in colorectal cancer survivors. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3464.
Abstract 3470: Obesity and metformin alter progression of triple negative breast cancer
Authors: Camacho Laura , Xue Lei , Dobrolecki Lacey , Hilsenbeck Susan G. , Nagi Chandandeep , Putluri Nagireddy , Lewis Michael T. , Sreekumar Arun , Jiralerspong Sao
Published: July 2016

Background: Epidemiological studies from us and others have reported an association of obesity with more aggressive breast tumors and >30% higher risk of recurrence and mortality compared to normal weight patients. This includes poorer outcomes for triple negative breast cancer (TNBC), an aggressive subtype with early recurrence and shorter survival. Metformin is a leading frontline therapy for type 2 diabetes and its safety profile is well documented. Laboratory and clinical studies including ours suggest that metformin may have anti-tumor effects in breast cancer. However, the nature and molecular mechanisms of these correlations are poorly understood.

Experimental design and methods: We used a PDX (patient-derived xenograft) breast cancer model in diet-induced obese mice to study how obesity promotes breast cancer growth and how metformin antagonizes these effects. After feeding the mice with a high-fat diet (HFD) vs. a control diet (CD), we transplanted a TNBC PDX into the mammary fat pads. When tumors reached about 50 mm3, mice in HFD and CD groups were randomly assigned to metformin treatment or no treatment. Tumor volumes were measured bi-weekly, and blood, tumors, and organs were collected after 4 weeks of treatment. Histological and immunohistochemical staining, serum adipokines array and metabolomics and lipidomics analyses were performed to examine potential mechanisms.

Results: HFD (obese) group compared to CD (lean) group was associated with larger tumors, whereas metformin compared to no treatment was associated with smaller tumors. A panel of adipokines and metabolites was identified to be differentially expressed between the groups.

Conclusion: Our results suggest a growth promoting effect of obesity in TNBC, while metformin appears to have the opposite effect. The identification of adipokines and metabolites involved in these effects will help us understand the mechanisms by which obesity and metformin alter the progression of TNBC.

Citation Format: Laura Camacho, Lei Xue, Lacey Dobrolecki, Susan G. Hilsenbeck, Chandandeep Nagi, Nagireddy Putluri, Michael T. Lewis, Arun Sreekumar, Sao Jiralerspong. Obesity and metformin alter progression of triple negative breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3470.
Abstract 3472: Does being married independently predict survival in patients with head and neck cancer? Results from a single institution
Authors: Osazuwa-Peters Nosayaba , Christopher Kara M. , Massa Sean T. , Cass Lauren , Walker Ronald J. , Varvares Mark A.
Published: July 2016

Purpose/Objectives

An important factor in cancer survivorship is social support, often coming from a spouse. There is an emerging literature on the role of spousal support in head and neck cancer (HNC) outcomes; however most are based on results from national databases. While more generalizable, these studies may not always capture the unique variation in different patient populations. This single institution study aimed to describe the association between marital status and outcomes of HNC; and to determine if marital status independently predicts survival in a local patient population.

Materials/Methods

We identified 460 patients aged 20 to 91 (59.31 ± 11.42) years diagnosed with squamous cell carcinoma of the head and neck at an academic tertiary referral center between 1997 and 2012 in this retrospective cohort study. Cox proportional hazards model assessed the effect of marital status on survival. Results are based on the final model constructed after accounting for covariates in the data.

Results

Our study population was made up of 73% males, and 82.2% Whites. We found an association between marital status and HNC survival. Unmarried HNC patients had a 66% increase in the hazard of death compared to married HNC patients (HR = 1.66, 95% CI = 1.23 - 2.23). This was after controlling for covariates, which included sociodemographic variables (age, race, sex, and health insurance status), social habits (tobacco and alcohol), primary anatomical subsite (oral cavity, oropharyngeal, laryngeal, and others), stage at presentation (early vs. late stage), and treatment modality (surgery, surgery with adjuvant therapies, other single modality therapy, and palliative care). Other factors found to be associated with an increased hazard of death were age (≥ 50 years), current tobacco use, late stage of presentation, palliative care, and laryngeal subsite.

Conclusion

Marital status is associated with head and neck cancer outcomes, and being married is an independent predictor of survival among patients. This result, found in previous national studies, held true in our local patient population. This underscores the need for the multidisciplinary HNC team to recognize this aspect of survivorship and to emphasize the need for social support among unmarried HNC patients. It could be that it is necessary to add social support to the clinical practical guidelines for managing head and neck cancer beyond palliative care.

Citation Format: Nosayaba Osazuwa-Peters, Kara M. Christopher, Sean T. Massa, Lauren Cass, Ronald J. Walker, Mark A. Varvares. Does being married independently predict survival in patients with head and neck cancer? Results from a single institution. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3472.
Abstract 3484: Analysis of in situ expression of hormone receptors and proliferation marker at a single cell level in breast cancer tissues
Authors: Ueno Takayuki , Isaka Hirotsugu , Itoh Hiroki , Miyamoto Kaisuke , Kitamura Manami , Imoto Shigeru
Published: July 2016

Background: Hormone receptors and proliferation markers are critical parameters for treatment selection of breast cancer patients. The expression of different parameters are currently assessed separately in different tissue sections but it is unclear how different parameters are co-expressed in a single cell in breast cancer tissues.

Samples and Methods: Breast cancer tissues from fifty-one patients with ER-positive HER2-negative breast cancer were analyzed. Expressions of ER and PgR were assessed in association with Ki67 using dual fluorescence immunohistochemistry with specific antibodies: SP-1 (abcam, Tokyo), 1E2 (Roche Diagnostics GmBH, Germany), and MIB1 (Dako Japan, Tokyo), respectively. More than 500 cancer cells were assessed in each tissue. Expression levels of each marker in a single cell were semi-quantitatively assessed by MetaMorph image analyzer (Molecular Devices Japan, Tokyo). All statistical analyses were performed using JMP ver.8.01 (SAS Institute Japan, Tokyo).

Results: To validate the system, Ki67 LI in breast cancer tissues were compared between this system and the regular DAB system. The two systems showed a good concordance (p < 0.0001). All cancer tissues expressed ER. There were two distinct populations among Ki67-positive proliferating cells according to PgR expression status: PgR-positive proliferating cells and PgR-negative proliferating cells. Since cell proliferation is regulated in cancer cells by at least two different drivers including hormone receptor and growth factor, it is conceivable that these two populations depend on different driving systems. Indeed, tissues with dominantly PgR-positive proliferating cells showed mostly histological grade 1 (15/20, 75%), while most of tissues with PgR-negative proliferating cells showed grade 2 and 3 (22/31, 71%) (p = 0.0025 by chi square test). Moreover, the multivariate analysis using the ordered logistic regression analysis showed that PgR status in proliferating cells is an independent factor associated with histological grade (p = 0.003) while PgR expression rate and Ki67 LI were not (p = 0.3 and 0.25, respectively).

Conclusion: PgR status in Ki67-positive proliferating cells is associated with histological grade in ER positive HER2-negative breast cancers. Our results suggest that different driving systems give different expression patterns of PgR and Ki67 at a single cell level, which may distinguish between luminal A and luminal B cancers.

Citation Format: Takayuki Ueno, Hirotsugu Isaka, Hiroki Itoh, Kaisuke Miyamoto, Manami Kitamura, Shigeru Imoto. Analysis of in situ expression of hormone receptors and proliferation marker at a single cell level in breast cancer tissues. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3484.
Abstract 3491: Rational combination of targeted agents and BH3 mimetics to improve cancer treatment
Authors: Montero Joan , Sadowicz Dorota E. , Ryan Jeremy , Yoon Charles H. , Haq Rizwan , Letai Anthony
Published: July 2016

Background: In the clinic is often observed that melanoma cancer patients first respond to targeted therapeutic agents like dabrafenib showing tumor burden shrinkage, but subsequently relapse with tumors that are more aggressive, no longer responsive to therapy and showing acquired metastatic capacity. Most chemotherapeutic agents kill via the mitochondrial pathway of apoptosis, but unfortunately tumors frequently are able to survive, often adapting their antiapoptotic strategy. We used a variant of the functional predictive test dynamic BH3 profiling (DBP) to determine the tumor's antiapoptotic defense adaptation over time, and rationally identify combination of therapies to restore programmed cell death.

Hypothesis: By using different BH3 peptides in DBP we can determine the adaptive tumor's antiapoptotic defense and adjust therapy to improve cancer cell death.

Results: In BRAF mutant melanoma, cKIT mutant GIST and HER2 overexpressing breast cancer cell lines, we performed DBP over time using different BH3 peptides: BIM, BAD, PUMA, MS1 and NOXA. We observed that when they are treated with specific targeted agents against their oncogenic drivers, there is an increase in MCL-1 dependence over time. This observation points to a possible tumor defense mechanism conferring resistance against anticancer drugs. Based on these observations, we exploited these altered dependencies by pretreating the cancer cells with these targeted agents and combining them with novel BH3 mimetics targeting MCL-1 to synergistically enhance cell death.

Exploiting DBP's capacity to measure changes in priming without the requirement for prolonged ex vivo culture, we assessed if this antiapoptotic evolution was also present in primary melanoma samples. Using FACS-based DBP to select for specific tumor populations, we tested the possible use of BH3 mimetics in combination with targeted agents to improve cancer treatment in the clinic.

Conclusions: Our preliminary results reveal the use of dynamic BH3 profiling to be used not only as a powerful real-time tool to predict chemotherapy response but also to evaluate the rational combination of targeted agents with BH3 mimetics in vitro and in the clinic.

[R.H. and A.L. contributed equally to this work.]

Citation Format: Joan Montero, Dorota E. Sadowicz, Jeremy Ryan, Charles H. Yoon, Rizwan Haq, Anthony Letai. Rational combination of targeted agents and BH3 mimetics to improve cancer treatment. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3491.
Abstract 3496: TRAF2 protects mammary epithelial and cancer cells from endoplasmic reticulum stress-induced apoptosis
Authors: Habelhah Hasem , Zhang Laiqun , Xialikaer Ainiwaer , Blackwell Ken
Published: July 2016

TRAF2 plays a key role in the immune response and cell survival by regulating the NF-κB and JNK pathways in response to most members of the TNF superfamily. TRAF2 has also been shown in cell culture systems to mediate IRE1α-induced activation of the ASK1-JNK pathway to promote apoptosis under conditions of acute endoplasmic reticulum (ER) stress. Gene knockout studies revealed that mice deficient for RANKL or NF-κB display defects in osteoclastogenesis and lactating mammary-gland development. Here, we report that TNFα and TRAF2 double knockout (DKO) mice develop with normal bone morphology and density, but the female mice display a severe lactation defect due to significantly increased apoptosis of cells in the lobuloalveolar tree. Unexpectedly, RANKL-induced activation of the canonical NF-κB pathway is not only normal in DKO mammary epithelial cells (MECs), but the expression of the NF-κB target genes is significantly enhanced due to constitutive activation of the non-canonical NF-κB pathway in the absence of TRAF2. On the other hand, cytotoxicity assays revealed that DKO MECs exhibit significantly increased susceptibility to apoptosis induced by chronic ER stress, but not by death receptor or DNA damage. In addition, siRNA-mediated knockdown of TRAF2 in breast cancer cells significantly sensitized the cells to chronic ER stress-induced apoptosis in a RIP1-dependent manner. These data suggest that the physiological function of TRAF2 is to protect the cells from apoptosis under conditions of pathologically relevant chronic ER stress. Given that the lactating MECs and rapidly growing cancer cells have high demands for protein syntheses and ER stress responses, our findings suggest that inhibition of the IRE1α-TRAF2 pathway could be used as a novel adjuvant therapy to treat rapidly proliferating cancer cells.

Citation Format: Hasem Habelhah, Laiqun Zhang, Ainiwaer Xialikaer, Ken Blackwell. TRAF2 protects mammary epithelial and cancer cells from endoplasmic reticulum stress-induced apoptosis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3496.
Abstract 350: Visualizing alectinib distribution in mice brain by using MALDI mass spectrometry imaging technique
Authors: Aikawa Hiroaki , Hayashi Mitsuhiro , Ryu Shoraku , Yamashita Makiko , Ohtsuka Naoto , Nishidate Masanobu , Hamada Akinobu
Published: July 2016

[Introduction]

In the development of anticancer drug, measurement of drug concentration in target tissue has been thought to be crucial for predicting its efficacy and safety. However, measuring homogenized tissue by using quantitative liquid chromatography tandem mass spectrometry (LC-MS/MS) completely loss the spatial information of the compound in a target tissue. Mass spectrometry imaging (MSI) has been recently applied as innovative tools to detect pharmaceutical distributions in heterogeneous targets; however, it still needs to be improved in quantitative capability or reproducibility.

[Purpose]

We propose quantitative approach for MSI (qMSI) that administere matrix-assisted laser desorption ionization (MALDI)-MSI, concurrently with LC-MS/MS by using serial sections. We applied qMSI for visualizing spatial distribution of alectinib, novel ALK inhibitor, in mouse brain and examined the effect of MDR1 (p-glycoprotein) on the pharmacokinetic profiles in FVB and Mdr1a/b knockout mice.

[Method]

Pharmacokinetic study of alectinib was conducted in FVB and Mdr1a/b KO mice. Plasma, cerebrospinal fluid (CSF) and brain tissue were collected at 1, 1.5, 2, and 4 hours after a single oral administration dose of 4 and 20 mg/kg. The concentration of alectinib in plasma, CSF and brain tissue sections was measured by LC-MS/MS. The distribution of alectinib in mice brain tissue section was visualized by qMSI that we established.

[Results]

Developing qMSI showed the brain distribution of alectinib in the range from 1.2 to 952.6 pg/mm2 per each section, and the results made with qMSI were in excellent agreement with the complementary measurements by using laser-micro dissection to verify the exposure level. While plasma alectinib concentrations had no difference between both mice, the diffuse alectinib distributions in brain were observed in Mdr1a/b knockout compared with that in FVB mice. The dominant localization of alectinib in choroid plexus and brain blood vessels of FVB mice was observed by 20 μm resolution MSI analysis.

[Conclusion]

Our qMSI technique is expected to contribute to the development of anticancer drugs and translational research by visualizing drug distribution in target organs and tumors.

Citation Format: Hiroaki Aikawa, Mitsuhiro Hayashi, Shoraku Ryu, Makiko Yamashita, Naoto Ohtsuka, Masanobu Nishidate, Akinobu Hamada. Visualizing alectinib distribution in mice brain by using MALDI mass spectrometry imaging technique. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 350.
Abstract 3500: Mitochondrially targeted p53 domains as a stand alone or adjunct to paclitaxel for the treatment of ovarian cancer
Authors: Lu Phong , Lim Carol S.
Published: July 2016

Although the main function of p53 is a nuclear transcription factor that has important roles in cell cycle arrest, DNA repair, and apoptosis, p53 can directly trigger the intrinsic apoptotic pathway through the mitochondria. p53 has been known to bind to mitochondrial anti-apoptotic proteins (Mcl-1, Bcl-2 and Bcl-XL) and pro-apoptotic proteins (Bak and Bax), which will cause the oligomerization of Bak and Bax. The result is the formation of permeable pores on the mitochondrial outer membrane, which in turn cause the release of cytochrome c and activation of caspase-3. Targeting p53 to the mitochondria is an attractive approach because it can cause a rapid apoptotic response. We have introduced the mitochondrial targeting signal (MTS) from Bak or Bax to the C-terminus of p53 and have shown the superior efficacy of p53-BakMTS and p53-BaxMTS over wtp53 in many human cancer cell lines. We have identified that the DNA binding domain (DBD) of p53 may be the minimal domain of p53 required for apoptosis. Our preliminary data in SKOV-3 ovarian cancer cells also suggests that DBD-BakMTS may work just as well as full length p53-BakMTS. In addition to BakMTS and BaxMTS, attaching pro-apoptotic factors such as Noxa or Bid to p53 has created a chimeric gene construct that show superior cell death activity than wide type p53. Our goal is to use a novel version of p53 directed to the mitochondria as a direct apoptogen to treat ovarian cancer, the most lethal gynecological malignancy with 69% of patients succumbing to this disease. Our next step will be to test these constructs in vivo using the syngeneic ID8 mouse model. To determine the activity of these mitochondrially targeted p53 constructs, TMRE, 7-AAD, and caspase 3/7 assays were performed in many human ovarian cancer cell lines including SKOV-3, OVCAR-3, Kuramochi, and mouse ovarian ID8 cells that have been transfected with our constructs. In all of our cell death and apoptotic assays, p53-Bax-MTS, p53-BakMTS, and p53-Noxa are always superior to wide type p53, regardless of the p53 status of the cells. Furthermore, we have also tested the possibility of combining our p53 constructs with paclitaxel, the current standard of care for ovarian cancer. Our p53 constructs showed a synergistic effect with paclitaxel and was able to reduce its IC5, indicating the possibility of dose-lowering of this highly toxic drug. Our ultimate goal is to use mitochondrially targeted p53 constructs that directly induce apoptosis alone or in combination with other chemotherapy drugs for the treatment of ovarian cancer.

Acknowledgements: NIH CA151847

Citation Format: Phong Lu, Carol S. Lim. Mitochondrially targeted p53 domains as a stand alone or adjunct to paclitaxel for the treatment of ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3500.
Abstract 3513: Promoting TRAIL apoptosis signaling using 17-beta-hydroxywithanolides
Authors: Brooks Alan D. , Xu Ya-ming , Wijeratne E. M. Kithsiri , Tewary Poonam , Henrich Curtis J. , Thomas Cheryl L. , Gunatilaka A. A. Leslie , Sayers Thomas J.
Published: July 2016

We have previously reported that withanolide E (WE), a steroidal lactone from Physalis peruviana, was highly active in sensitizing various human carcinoma cell lines to TRAIL-mediated apoptosis. Treatment of cancer cells with WE induced a reduction in the levels of the antiapoptotic proteins cFLIPL and cFLIPS, resulting in an increased activation of caspase-8 on subsequent TRAIL binding to its death receptors DR4 or DR5. The reduction in cFLIPL and cFLIPS was due to their destabilization and increased degradation by the proteasome. Interestingly, WE (a 17-beta-hydoxywithanolide, 17-BHW) was a far superior TRAIL sensitizer than more widely studied withaferin A (WFA) and its analogues, which lack the 17-beta-hydroxy group and bear an opposite side chain orientation, exhibit more promiscuous reactivity and are much more directly toxic to cells. Therefore, over 30 natural and semi-synthetic 17-BHWs were evaluated for their ability to promote death ligand-mediated cancer cell death. The 17-BHWs used in this work were obtained by the application of an efficient method of plant biomass production involving our innovative and patented soil-less aeroponic cultivation of P. crassifolia and P. peruviana and by chemical modification of natural withanolides produced by these plants. Our studies identified several 17-BHWs that were 4-8 fold more potent than WE in sensitizing the renal carcinoma cells ACHN to TRAIL-mediated apoptosis. These more active 17-BHWs were also more efficient at reducing cellular levels of cFLIPL and cFLIPS and enhancing caspase-8 activation. Preliminary structure activity relationship (SAR) studies suggested that the enone moiety in ring A was essential for activity. In addition acetoxylation at C-18, an alpha orientation of the lactone group and the double bond at C-24(25) of the lactone ring played important roles in determining the activity of 17-BHWs as TRAIL sensitizers. This suggests that the 17-BHW scaffold is amenable to optimization by a medicinal chemistry approach, which could lead to the identification of highly active natural product-based sensitizers of cancer cells to TRAIL-mediated apoptosis. The cellular molecular target(s) of active 17-BHWs are currently under further investigation.

Funded by FNLCR Contract HHSN261200800001E

Citation Format: Alan D. Brooks, Ya-ming Xu, E. M. Kithsiri Wijeratne, Poonam Tewary, Curtis J. Henrich, Cheryl L. Thomas, A. A. Leslie Gunatilaka, Thomas J. Sayers. Promoting TRAIL apoptosis signaling using 17-beta-hydroxywithanolides. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3513.
Abstract 3515: MiR-320b-regulated TRIAP1 prevents mitochondria fragmentation and apoptosis by controlling cytochrome c release in nasopharyngeal carcinoma
Authors: Li Ying-Qin , Liu Na , Ma Jun
Published: July 2016

Mitochondria play critical roles in apoptosis. Imbalance of mitochondria fusion/fission leads to mitochondrial fragmentation and initiates intrinsic apoptosis, but the regulation of mitochondrial fragmentation in nasopharyngeal carcinoma (NPC) remains undefined. Here we show that TRIAP1 is aberrantly overexpressed and associated with poor survival in NPC patients. TRIAP1 overexpression promotes cell growth and suppresses NPC cell death in vitro and in vivo, whereas TRIAP1 knockdown inhibits cell tumorigenesis and enhances apoptosis through induction of mitochondrial fragmentation, membrane potential alteration and release of cytochrome c from mitochondria to cytosol. In intersection with our previous miRNA data and available bioinformatics algorithms, we identified miR-320b as a negative regulator of TRIAP1 expression. Further studies show that miR-320b suppresses NPC cell proliferation and enhances mitochondrial fragmentation and apoptosis by targeting TRIAP1. Moreover, loss of miRA-320b expression is conversely correlated with TRIAP1 overexpression and poor prognosis in NPC patients. This study reveals the regulation of miR-320b/TRIAP1 in mitochondrial fragmentation and apoptosis, and suggests potential therapeutic targets for NPC treatment.

Citation Format: Ying-Qin Li, Na Liu, Jun Ma. MiR-320b-regulated TRIAP1 prevents mitochondria fragmentation and apoptosis by controlling cytochrome c release in nasopharyngeal carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3515.
Abstract 3520: Solidago virga-aurea extract induced apoptosis of breast cancer cells through FOXO3 mediated bim expression
Authors: Kim Haesung , Kim Chea Ha , Kim Jeong-Hyeon , Lee Jeong-Min , Jeon Jeong-Ho , Lee Jae-Yong
Published: July 2016

Solidago virga-aurea (European goldenrod or woundwort) is an herbaceous perennial plant of the family Asteraceae. It is used for medical system with astringent, diuretic, antiseptic and homeopathic properties. And the studies about Solidago virga-aurea are in progress for the other functions.

Recently, we found that Solidago virga-aurea extract induces apoptosis of breast cancer cells. MTT assay showed that cellular viability was decreased in Solidago virga-aurea extract treated MDA-MB-231 and MCF-7 breast cancer cell lines by dose and time dependent manner. And apoptotic protein, especially bim, was increased in Solidago virga-aurea extract treated MCF-7 cell line. However, anti-apoptotic protein, Bcl-xL, does not changed by Solidago virga-aurea extract. In order to find out positive regulator of bim expression, we tested whether FOXO (forkhead-box transcription factor, group O) 3a and AMPK (5’ AMP-activated protein kinase) protein expression. In immunoblot analysis showed that FOXO3a expression level was dramatically increased in Solidago virga-aurea treated MCF-7 cells without changes of AMPK expression. Moreover, immunostaining showed that Solidago virga-aurea induces co-localization of FOXO3a and bim proteins in MCF-7 and MB-231 cells.

Taken together, Solidago virga-aurea induces apoptosis by FOXO3a mediated up-regulation of bim protein. The mechanisms of Solidago virga-aurea mediated FOXO3a regulation is not fully elucidated yet, but further studies about this molecular mechanism between two proteins will give some ideas for cancer therapies for certain breast cancer patients.

Citation Format: Haesung Kim, Chea Ha Kim, Jeong-Hyeon Kim, Jeong-Min Lee, Jeong-Ho Jeon, Jae-Yong Lee. Solidago virga-aurea extract induced apoptosis of breast cancer cells through FOXO3 mediated bim expression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3520.
Abstract 3522: Cyclins and CDKs regulation and caspase cascade activation by cucurbitacin D induced cell cycle arrest and apoptosis in pancreatic tumor
Authors: Kim Myeong-Sun , Kim Ji Hye , Ku Jin Mo , Hong Se Hyang , Lee Kangwook , Choi Hyeong Sim , Woo Sang Mi , Chang Jee Yun , Kim Tai Young , Ko. Seong Gyu Ko
Published: July 2016

Pancreatic cancer has a poor prognosis and very low survival rate over the world. Because pancreatic cancer probably is diagnosed at a late stage, aggressive local invasion, and poor response to chemotherapy. Gemcitabine was the standard treatment for advanced and metastatic pancreatic cancer patients, but it is associated with multiple adverse effects-fever, fatigue, nausea, and drug resistance. Whether cucurbitacin D has any efficacy against human pancreatic cancer was examined in cell culture system. In vitro, cell viability was measured by MTT assay to recognize of cell cytotoxicity. Consequently, cytotoxicity was observed at a low concentration of cucurbitacin D. Wound healing assay and clonogenic assay indicated that cucurubitacin D inhibited the growth of cell growth through cyclins and CDKs regulation, and decreased colony-forming ability. Also, this compound down-regulated expression level of anti-apoptotic protein, Bcl-2, up-regulated of pro-apoptotic molecule Bax, and activated caspase-8, caspase-3 cascade extrinsic pathway. Additionally, PARP, caspase-3 substrate, protein was cleaved by cucurbitacin D treatment. Overall, our study suggest that cucurbitacin D could be a clinical medicine for the treatment of pancreatic cancers.

Citation Format: Myeong-Sun Kim, Ji Hye Kim, Jin Mo Ku, Se Hyang Hong, Kangwook Lee, Hyeong Sim Choi, Sang Mi Woo, Jee Yun Chang, Tai Young Kim, Seong Gyu Ko Ko. Cyclins and CDKs regulation and caspase cascade activation by cucurbitacin D induced cell cycle arrest and apoptosis in pancreatic tumor. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3522.
Abstract 3533: Role of autophagy-related protein expression in patients with rectal cancer treated with neoadjuvant chemoradiotherapy
Authors: Ko Yoon Ho , Hong Soon Uk , Sun Der Sheng , Won Hye Sung , Lee Myung Ah , Shim Byoung Young
Published: July 2016

Background: Autophagy, a cellular degradation process, has complex roles in tumourigenesis and resistance to cancer treatment in humans. The aim of this study was to explore the expression levels of autophagy-related proteins in patients with rectal cancer and evaluate their clinical role in the neoadjuvant chemoradiotherapy setting.

Methods: All specimens evaluated were obtained from 101 patients with colorectal cancer who had undergone neoadjuvant chemoradiotherapy and curative surgery. The primary outcomes measured were the expression levels of two autophagy-related proteins (microtubule-associated protein 1 light chain 3 beta (LC3β) and beclin-1) by immunohistochemistry and their association with clinicopathological parameters and patient survival.

Results: Among the 101 patients, the frequency of high expression of beclin-1 was 31.7% (32/101) and that of LC3β was 46.5% (47/101). A pathologic complete response was inversely associated with LC3β expression (P = 0.003) and alterations in the expression of autophagy-related proteins (P = 0.046). In the multivariate analysis, however, autophagy-related protein expression did not show prognostic significance for relapse-free survival or overall survival.

Conclusions: High expression of autophagy-related proteins shows a strong negative association with the efficacy of neoadjuvant chemoradiotherapy in patients with rectal cancer. Autophagy has clear implications as a therapeutic target with which to improve the efficacy of neoadjuvant chemoradiotherapy.

Citation Format: Yoon Ho Ko, Soon Uk Hong, Der Sheng Sun, Hye Sung Won, Myung Ah Lee, Byoung Young Shim. Role of autophagy-related protein expression in patients with rectal cancer treated with neoadjuvant chemoradiotherapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3533.
Abstract 3538: Effects of oral chloroquine administration on a preclinical mouse model of PTEN/p53-deficient prostate cancer
Authors: De Velasco Marco A. , Sugimoto Koichi , Kura Yurie , Hatanaka Yuji , Yamamoto Yutaka , Oki Takashi , Yoshimura Kazuhiro , Nozawa Masahiro , Yoshikawa Kazuhiro , Nishio Kazuto , Uemura Hirotsugu
Published: July 2016

Autophagy is cellular process that can promote tumor survival during periods of metabolic and therapeutic stress. Inhibition of autophagy can promote senescence resulting in decreased tumor growth. Genetic alterations of PTEN and p53 are frequently present in advanced human prostate cancer and are associated with poor prognosis. In mouse models, the inactivation of PTEN leads to the development of prostate tumors that progress slowly, in part due to the induction of PTEN-(loss)-induced cellular senescence. Concomitant inactivation of both PTEN and P53 in prostate leads to the development of an aggressive cancer phenotype that can overcome PTEN-(loss)-induced cellular senescence and is characterized with accelerated growth, metastatic development and decreased survival. Published findings show that in some cancer models, the effects of autophagy inhibition are dependent on p53 status. In this study we assessed the effects of pharmacological inhibition of autophagy with chloroquine (CQ) on the growth and progression of mouse prostate tumors driven by the concomitant inactivation of PTEN and p53. CQ is a lysosomotropic agent that has widely used to impede autophagy and inhibit the growth of human tumor cells in vitro and in vivo. Genetically engineered mice with the conditional inactivation of PTEN and p53 were used to evaluate the antitumor effects and therapeutic benefit of CQ in a drug intervention model of prostate cancer. Twenty-seven-week-old PTEN/p53 double knockout mice bearing prostate tumors were randomized to control and oral CQ (100 mg/kg in drinking water) treatment groups. Progression-free survival was 145 days versus 161 days (P = 0.703) for control and CQ treated mice, respectively. Median overall survival was 170 days versus 147 days (P = 0.523) for control and CQ treated mice, respectively. Tumor burden assed by the median genitourinary tract weight was 2.42 grams versus 2.19 grams (P = 0.645) for control and CQ treated mice, respectively. Our molecular assays revealed that CQ failed to consistently inhibit autophagy in tumors at the present dose. Overall, our findings show that CQ alone did not provide treatment benefit in mouse prostate cancer with loss of p53.

Citation Format: Marco A. De Velasco, Koichi Sugimoto, Yurie Kura, Yuji Hatanaka, Yutaka Yamamoto, Takashi Oki, Kazuhiro Yoshimura, Masahiro Nozawa, Kazuhiro Yoshikawa, Kazuto Nishio, Hirotsugu Uemura. Effects of oral chloroquine administration on a preclinical mouse model of PTEN/p53-deficient prostate cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3538.
Abstract 3539: Vitamin D/EB 1089 mediated radio-sensitization of lung cancer cells and head and neck cancer cells
Authors: Thekkudan Theresa , Sharma Khushboo , Saleh Tareq , Gewirtz David
Published: July 2016

Lung cancer is a principal factor contributing to cancer-related mortality in the United States. Patients with lung cancer generally have a poor prognosis and resection is often not possible. Our previous work has demonstrated that Vitamin D and its analog, EB1089 (EB), enhance radio sensitivity of H460 and A549 non-small cell lung cancer (NSCLC) cells (Sharma et al; A novel cytostatic form of autophagy in sensitization of non-small cell lung cancer cells to radiation by vitamin D and the vitamin D analog, EB 1089. Autophagy; 2014;10(12):2346-61). In this study, we have extended these findings of Vitamin D/EB 1089 induced radio-sensitization in multiple tumor cell lines such as Lewis Lung mouse non-small cell lung cancer cells (LLC), head and neck cancer cells (HN30) and a primary cell line established from a lung tumor obtained from a cancer patient.

All cell lines were treated with multiple doses of radiation in the range of 2-6 Gy and EB1089 was used at concentration of 100nM. Sensitivity was determined based on viable cell number and clonogenic survival.

We observed enhanced radiation sensitivity of LLC and HN30 cells with the combination treatment of radiation and EB1089. However, as reported previously by our laboratory (Sharma et al., 2014; Autophagy), sensitization was dependent on the presence of functional p53 and was not evident in HN6 head and neck tumor cells that are mutant in p53. These data suggest that Vitamin D or a Vitamin D analog could be used to enhance radiation sensitivity of various malignancies but only where the tumors express wild type p53. Furthermore, we were able to demonstrate sensitization in a primary cell line established from lung tumor tissues utilizing conditioned medium from stromal cells treated with EB1089. These data suggest that stromal re-programming of cancer cells could influence radiation sensitivity.

In studies to define the signaling pathways associated with radiation sensitization, we also observed inactivation by EB 1089 of the NF-kB pathway that was activated by radiation alone in H460 and H1299 NSCLC cells. Silencing of AMPK attenuated radiation sensitization by EB 1089. Sensitization was accompanied by enhanced secretion of the autophagy/senescence related marker, HMGB1. Taken together, these studies indicate that Vitamin and/or its analogs enhance radiosensitivity by cellular pathways involving NF-kB and AMPK and that HMGB1 may be secreted to activate an immune response to the combination treatment.

Citation Format: Theresa Thekkudan, Khushboo Sharma, Tareq Saleh, David Gewirtz. Vitamin D/EB 1089 mediated radio-sensitization of lung cancer cells and head and neck cancer cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3539.
Abstract 3547: CEBPD induces apoptosis through autophagy in metformin-treated HCC
Authors: Lin. I P.
Published: July 2016

The patients with hepatocellular carcinoma (HCC) face the risk of cancer relapse or liver dysfunction. Metformin (Met), a first-line drug of type 2 diabetes, has been proved that it could induce G1 arrest and prevent or inhibit HCC. Previously, we demonstrated that the transcription factor CCAAT/enhancer-binding protein delta (CEBPD) plays a tumor suppressor and is responsive to many anticancer drugs in HCC. In this study, we found that Met could induce autophagy, growth arrest and further apoptosis in Huh7 cells. Met enhanced CEBPD expression via stabilizing its protein stability. The loss of CEBPD attenuated the Met-induced autophagy, growth arrest and apoptosis. Importantly, in addition to increasing P27 and down-regulating cyclin D1, CEBPD also contributed the increases of LC3B and ATG3 and LC3 puncta formation. We revealed a novel involvement of CEBPD in Met-induced growth arrest, apoptosis and autophagy of liver cancer cells through regulating p27, cyclinD1, LC3B and ATG3.

Citation Format: I P. Lin. CEBPD induces apoptosis through autophagy in metformin-treated HCC. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3547.
Abstract 3548: In vivo CRISPR reveals dual activation of apoptosis and necroptosis as means to eradicate drug resistant leukemia
Authors: Aguadé-Gorgorió Júlia , McComb Scott , Cario Gunnar , Stanulla Martin , Eckert Cornelia , Bourquin Jean-Pierre , Bornhauser Beat C.
Published: July 2016

Dysregulation of apoptosis constitutes a general mechanism for refractory acute lymphoblastic leukemia (ALL) to escape cell death during chemotherapy. We have assessed the potential of SMAC mimetic compounds (SM) to eradicate drug resistant ALL by blocking the pro-survival cellular inhibitor of apoptosis proteins (cIAPs). On the basis of a large xenograft bank of primary ALL samples, obtained by xenotransplantation in immunodeficient NSG mice, we detected exquisite sensitivity to SM in vitro in a third of the cases tested, including drug-resistant and relapsed samples. Potent in vitro activity was highly predictive of in vivo efficacy in the xenograft model. Treatment of transplanted mice with SM not only delayed leukemia progression, but also completely eliminated leukemia burden in established disease for several weeks following treatment. SM activity required the presence of RIP1, since lentiCRISPR mediated gene knockout of RIP1 in patient-derived ALL cells blocked SM-induced cell death in vivo and in vitro. In contrast, RIP1 loss had no significant impact on the response to standard antileukemic therapies, supporting a view that RIP1-dependent death pathways are not likely to be selected against by front line chemotherapy. Furthermore, using a multicolor lentiCRISPR approach for simultaneous disruption of multiple genes, we found that downstream of RIP1, the response to SM was dependent on simultaneous execution of apoptosis and necroptosis, since genetic targeting of both cell death pathways was required to block SM-induced death. Upstream of RIP1, expression levels of TNF receptor II (TNFR2) in primary ALL correlated with sensitivity to SM, and genetic disruption of TNFR2 conferred resistance to SM treatment in vivo. Thus, SM treatment circumvents drug resistance through parallel activation of RIP1-dependent apoptosis and necroptosis, providing a strong rationale for application in salvage treatment of refractory childhood ALL.

Citation Format: Júlia Aguadé-Gorgorió, Scott McComb, Gunnar Cario, Martin Stanulla, Cornelia Eckert, Jean-Pierre Bourquin, Beat C. Bornhauser. In vivo CRISPR reveals dual activation of apoptosis and necroptosis as means to eradicate drug resistant leukemia. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3548.