bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2253: Lymphangiogenesis augments the anti-tumor efficacy of radiotherapy in mouse melanoma
Authors: Mitrousis Nikolaos , Sasso Maria Stella , Weichselbaum Ralph R. , Hubbell Jeffrey A. , Swartz Melody A.
Published: September 2020

Radiotherapy (RT) constitutes a significant component of cancer therapy in the clinic, with approximately 50% of all cancer patients receiving treatment over the course of their disease. In addition to acting as a DNA-damaging agent, RT leads to the release of tumor antigens, damage-associated molecular patterns, and cytokines, a process termed immunogenic cell death. This phenomenon triggers the initiation of anti-tumor immune responses, which have been shown to be essential for the complete therapeutic response to RT. Despite its diverse mechanisms of action and broad applicability, RT does not typically lead to curative responses and combinations with other treatment modalities such as immunotherapy are being explored to improve efficacy. Our laboratory and others have previously established that tumor-associated lymphatics can actively modulate anti-tumor immune responses and potentiate immunotherapy. Here, we asked whether lymphangiogenesis influences the immune response and anti-tumor efficacy elicited by RT. We employed two variants of the aggressive B16F10 melanoma model: a variant that was transduced to overexpress the lymphangiogenic factor VEGF-C (B16-VEGF-C), and a mock-transduced (B16-ctrl) variant. Tumor-bearing animals were treated by ablative doses of localized RT and monitored for tumor growth and survival, while anti-tumor immune responses were assessed by flow cytometry and ELISA. We observed that the therapeutic efficacy of RT was greater in B16-VEGF-C melanomas compared to B16-ctrl, as evidenced by a significant inhibition of tumor growth and extended animal survival. In line with this finding, B16-VEGF-C melanomas exhibited a significantly higher CD8+ T cell infiltration and activation upon ex vivo restimulation after RT. These responses were partially inhibited by a function-blocking antibody of the main VEGF-C receptor, VEGFR3. To gain further insight, we assessed the tumor infiltration and activation status of antigen-presenting cells. Consistent with the observed T cell response, we found that both dendritic cells and macrophages exhibited a greater expression of co-stimulatory molecules in B16-VEGF-C melanomas after RT. Together, our findings demonstrate that lymhangiogenesis may be a key contributing factor underlying tumor responsiveness to RT.

Citation Format: Nikolaos Mitrousis, Maria Stella Sasso, Ralph R. Weichselbaum, Jeffrey A. Hubbell, Melody A. Swartz. Lymphangiogenesis augments the anti-tumor efficacy of radiotherapy in mouse melanoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2253.
Abstract 2276: Preclinical development and mechanism of action studies of NM21-1480, a PD-L1/4-1BB/HSA trispecific MATCH3 therapeutic clinical candidate
Authors: Snell Daniel , Gunde Tea , Warmuth Stefan , Lichtlen Peter , Tietz Julia , Brock Matthias , Simonin Alexandre , Hess Christian , Christopher Weinert , Heiz Robin , Flueckiger Naomi , Zeberer Julia , Diem Dania , Mahler Dana , Morenzoni Diego , Wickihalder Belinda , Muntwiler Simone , Poelderl Antonia , Kuettner Benjamin , Wagen Sandro , Meyer Sebastian , Egan Timothy , Urech David
Published: September 2020

Antagonistic molecules targeting the PD-1/PD-L1 axis have shown excellent activity in the clinic. However the majority of patients do not respond to the therapy due to multifaceted reasons implicating a non-effective activation of the immune system in those patients. The co-stimulatory molecule 4-1BB has been shown to be a key signalling component of T cells and the combination of 4-1BB activation and PD-1/PD-L1 antagonism has been shown to be highly active in preclinical models. Systemic application of first generation anti-4-1BB antibodies however have resulted in dose limiting hepatic toxicities. We have generated a novel 4-1BB/PD-L1/HSA trispecific MATCH3 immunomodulatory drug candidate (NM21-1480) that agonizes 4-1BB conditionally upon PD-L1 binding / blockade. Here we show the preclinical development data package for NM21-1480 demonstrating exquisite tumour-specific T cell activation associated with both 4-1BB agonism as well as PD-L1/PD-1 antagonism. We also demonstrate tumour specific localisation and accumulation of NM21-1480 in a mouse xenograft model. We report on the toxicity profile and pharmacokinetic properties of the molecule in non-human primates. We plan to initiate a first-in-human clinical study in the second half of 2020 to determine the safety, tolerability and first signs of clinical activity of the molecule.

Citation Format: Daniel Snell, Tea Gunde, Stefan Warmuth, Peter Lichtlen, Julia Tietz, Matthias Brock, Alexandre Simonin, Christian Hess, Weinert Christopher, Robin Heiz, Naomi Flueckiger, Julia Zeberer, Dania Diem, Dana Mahler, Diego Morenzoni, Belinda Wickihalder, Simone Muntwiler, Antonia Poelderl, Benjamin Kuettner, Sandro Wagen, Sebastian Meyer, Timothy Egan, David Urech. Preclinical development and mechanism of action studies of NM21-1480, a PD-L1/4-1BB/HSA trispecific MATCH3 therapeutic clinical candidate [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2276.
Abstract 2322: A polygenic risk score-based genetic stratification for endometrial cancer
Authors: Bafligil Cemsel , Thompson Deborah J. , Lophatananon Artitaya , Smith Miriam J. , Dennis Joe , Evans Gareth D. , Crosbie Emma J.
Published: September 2020

Endometrial cancer (EC) is the most common of gynecological cancers in the UK. There is evidence for genetic predisposition to EC, however, single nucleotide polymorphisms have not been considered in risk prediction models so far. We hypothesized that a polygenic risk score (PRS) based on a panel of robust SNPs may be useful to predict women at high risk of developing EC. We conducted a systematic review to identify the most robust SNPs which may be associated with EC risk. The resulting panel of SNPs was validated in an independent cohort comprising EC cases, and control women with intact uterus. In the present study, we used 560 cases and 1202 controls of broad-European origin to construct a PRS. Our systematic review suggested twenty four SNPs, nineteen of which were reported to reach genome-wide significance in large GWAS. Preliminary results using 560 cases and 1202 controls produced effect sizes, directions and dosages similar to published data. The mean PRS for EC cases was 0.64 (SD=0.43, 95%CI 0.60-0.68) while mean PRS for controls was 0.48 (SD=0.42, 95% CI 0.46-0.51). The AUC of the PRS achieved was 0.61. Women in the highest tertile of the PRS score had a 2.4 times increased risk of developing EC compared to women in the lowest tertile (OR=2.40, 95%CI 1.85-3.09, p = 1.47E-11). Women in the mid tertile also had a modestly increased risk of EC compared to the lowest tertile (OR=1.40, 95%CI 1.06-1.84, p = 0.017). There was a clear trend of increasing risk with category (p = 6.25E-12). There is mounting evidence for polygenic input from common variants in EC susceptibility. Our results, for the first time, validate the potential use of a PRS for risk stratification of women at high risk of EC. When considered alongside epidemiological risk factors, the PRS could provide a good discrimination for early detection and prevention of EC in a clinical setting.

Citation Format: Cemsel Bafligil, Deborah J. Thompson, Artitaya Lophatananon, Miriam J. Smith, Joe Dennis, Gareth D. Evans, Emma J. Crosbie. A polygenic risk score-based genetic stratification for endometrial cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2322.
Abstract 233: Tumor-associated carbonic anhydrase IX maintains cellular proliferation by regulating tumor metabolism: a novel link revealed by proteomics
Authors: Benej Martin , Svastova Eliska , Banova Radivojka , Kopacek Juraj , Gibadulinova Adriana , Kery Martin , Arena Simona , Scaloni Andrea , Vitale Monica , Zambrano Nicola , Papandreou Ioanna , Denko Nicholas C. , Pastorekova Silvia
Published: September 2020

Tumor hypoxia is a critical microenvironmental stress occurring in virtually all types of solid tumors. Survival of cancer cells is dependent upon their ability to adapt to these limiting conditions. Hypoxia-induced carbonic anhydrase IX (CA IX) plays a crucial adaptive role in response to hypoxic and acidic microenvironments by catalytically hydrating extracellular CO2 to produce bicarbonate ions that maintain intracellular pH (pHi). In order to determine how CAIX function impacts other cellular processes, we used proteomic profiling to identify changes in response to transient CA IX knockdown in hypoxia. We discovered a significant reduction of key glycolytic enzyme levels with a most prominent decrease in lactate dehydrogenase (LDH). Our data shows that in addition to participating in cellular regulation of pHi, CA IX significantly stimulates glycolytic flux, lactate production and rate of proliferation in hypoxia. Because LDH activity contributes to pHi regulation by consuming cytoplasmic protons in the process of NADH regeneration, CA IX knockdown-mediated LDH suppression may further compromise the pHi regulating ability in hypoxia. Interestingly, addition of the alternative LDH substrate alpha-ketobutyrate compensates for reduced lactate production and restores pHi and proliferation in CA IX-deficient cells. These findings suggest that CA IX confers adaptive plasticity and supports proliferation of cancer cells both directly and indirectly - through its effect as a pHi regulator and by maintaining glycolysis-permissive intracellular environment, respectively. Because CA IX expression is limited almost exclusively to tumors, our findings highlight the role as well as provide mechanistic insight for the utility of CA IX as an anti-cancer target and suggests that highly glycolytic tumors might be particularly vulnerable to CA IX-targeted therapy.

Citation Format: Martin Benej, Eliska Svastova, Radivojka Banova, Juraj Kopacek, Adriana Gibadulinova, Martin Kery, Simona Arena, Andrea Scaloni, Monica Vitale, Nicola Zambrano, Ioanna Papandreou, Nicholas C. Denko, Silvia Pastorekova. Tumor-associated carbonic anhydrase IX maintains cellular proliferation by regulating tumor metabolism: a novel link revealed by proteomics [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 233.
Abstract 2332: Correlation of lifestyle factors with epigenetic aging unveiled in the practice of a DNA methylation-based age test
Authors: Guo Wei , Chew Yap Ching , Yang Xiaojing , Suwoto Michiko , Yamada Taikun , Jia Xi Yu
Published: September 2020

Background: DNA methylation clocks have been developed recently to estimate epigenetic age which is associated with chronological age in mouse and human. Changes in epigenetic age were reported in patients with health conditions including cancer. Different from genetic variations, epigenetic properties can be modified by environmental factors. Epigenetic age tests using blood samples have been offered directly to customers recently. We sought to analyze relationships between epigenetic aging and lifestyle factors, medication and aging interventions in the hope of assisting healthier living.

Methods: Epigenetic age, called DNAge, is estimated using linear regression of DNA methylation levels at 206 loci containing 1344 CpGs and chronological age. Delta age is defined as the difference of epigenetic age minus chronological age. Correlations between lifestyle factors or medication and delta age of hundreds of whole blood samples were evaluated using biweight midcorrelation. Statistical significance is set at 0.05.

Results: Our results showed positive correlations between accelerated epigenetic aging and BMI (bicor=0.148, p=0.009, n=311), current smoker (bicor=0.143, p=0.011, n=315), or stress (bicor=0.088, p=0.128, n=302). Factors with positive correlation but not reaching statistical significance included total cholesterol, alcohol consumption and Keto diet. Factors with negative correlation but not statistically significant included exercise, balanced diet, mostly meat, mostly vegetables, and education.

Conclusions: The epigenetic age analysis of real customer data confirmed beneficial and adversarial health effects of certain lifestyle factors. The DNAge test offered proves to be useful as a biomarker for biological age and as a practical tool for monitoring and evaluation of health status which may be used in cancer epidemiology study and help cancer prevention.

Citation Format: Wei Guo, Yap Ching Chew, Xiaojing Yang, Michiko Suwoto, Taikun Yamada, Xi Yu Jia. Correlation of lifestyle factors with epigenetic aging unveiled in the practice of a DNA methylation-based age test [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2332.
Abstract 2335: Heterozygous PALB2 mutations cause replication stress and DNA repair defects in carrier derived lymphoblastoid cell lines
Authors: Laurila Niina , Borgmann Kerstin , Bose Muthiah , Meyer Felix , Pylkäs Katri , Xia Bing , Foulkes William , Southey Melissa , Pospiech Helmut , Winqvist Robert
Published: September 2020

During the last decade, PALB2 has been defined as a high-risk breast cancer susceptibility gene alongside BRCA1 and BRCA2. Heterozygous mutation in PALB2 increases the lifetime breast cancer risk of female carriers to an average of 53%, but the risk estimate is affected by family history, studied population, and the specific pathogenic variant.

We have previously shown that lymphoblastoid cell lines (LCLs) of persons heterozygous for PALB2 c.1592delT, a Finnish founder mutation, show defects in DNA replication and damage response. Now we examine how various heterozygous, protein-truncating PALB2 founder mutations from different populations affect cell function, and whether the observed phenotype correlates with disease risk. The study cohorts come from Canada, Australia, and Finland and consist of 80 LCLs of PALB2 mutation carriers (c.1592delT, c.2323C>T, c.3113G>A or c.3323delA) and suitable population controls. There is some evidence that these variants are associated with different magnitudes of breast cancer risk, and PALB2 c.3113G>A has been associated with an exceptionally high cancer risk of 91% (95% CI 44-100) to age 70.

Utilizing these LCLs, we have studied DNA damage response, cell cycle progression, and DNA replication with various methods. DNA fiber assay results show that PALB2 mutation carriers elongate their DNA more slowly during replication, likely due to increased fork stalling, but compensate this by increased dormant origin firing. The strongest replication phenotype was observed for PALB2 c.3113G>A carriers. High content analysis shows that most of the mutation carriers had compromised RAD51 foci formation after DNA damage, whereas 53BP1 foci levels were elevated. These results indicate that during DNA double-strand break repair, heterozygous PALB2 mutation carriers may rely more on alternative, error-prone pathways that are independent of RAD51, thus contributing to genomic instability. Additionally, we are currently performing RNA-Seq to determine if the LCLs show adaptation of their gene expression in response to pathogenic PALB2 variants.

Citation Format: Niina Laurila, Kerstin Borgmann, Muthiah Bose, Felix Meyer, Katri Pylkäs, Bing Xia, William Foulkes, Melissa Southey, Helmut Pospiech, Robert Winqvist. Heterozygous PALB2 mutations cause replication stress and DNA repair defects in carrier derived lymphoblastoid cell lines [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2335.
Abstract 2345: Statin use associated with reduced risk of hepatocellular carcinoma recurrence following liver resection: A systematic review and meta-analysis
Authors: Yeung Chun Philip , Cheung Siu Tim , Ng Kwok Chai Kelvin , Lai Bo San Paul , Chong Ching Ning Charing
Published: September 2020

The effect of statins in preventing hepatocellular carcinoma (HCC) recurrence after curative liver resection has been controversial. Some retrospective studies identified perioperative use of statins was associated with significantly lower risk of HCC recurrence and improve recurrence-free survival (RFS) after liver resection, among patients with or without chronic hepatitis viral infection in multivariate survival analysis. However, a recent single-center study enrolled 430 cases in Taiwan revealed that the protective effect of statins was only significant in univariate analysis but not significant in multivariate analyses. To evaluate the effect of statins on the risk of HCC recurrence after curative liver resections, we performed a systematic review and meta-analysis on this topic. A systematic search of Medline, Embase, and Web of Science was conducted through December 2019. Studies were included if they evaluated perioperative exposure to statins, reported the recurrence of HCC after curative resection, and reported hazard ratios (HR) of multivariable analysis by Cox proportional hazards model. Summary HR estimates with 95% confidence intervals (CI) were calculated using the random-effects model. The analysis included 5 studies reporting the recurrence of HCC among 8,586 patients after liver resection, in which 411 of them received perioperative statins treatments. A meta-analysis of the studies showed a significant (39%) reduction in the risk of HCC recurrence among patients who took statins (adjusted HR, 0.61; 95% CI, 0.49-0.76), with no or little heterogeneity among studies (χ2=5.58, df=4, I2=28%). In conclusion, perioperative statins use may improve recurrence-free survival and decrease the risk of HCC recurrence after curative liver resection. Further prospective studies and larger randomized controlled trials (RCTs) are warranted to validate the conclusion.

Statin usersNon-statin usersAdjusted Hazard RatioStudyTotalTotalWeightIV, Random, 95%CIYear of publicationWu 2012175439437.6%0.68 (0.53-0.87)2012Lee 2016132207831.2%0.66 (0.49-0.90)2016Kawaguchi 2017317037.0%0.34 (0.12-0.75)2017Nishio 20184360014.5%0.42 (0.25-0.71)2018Young 2019304009.8%0.79 (0.41-1.52)2019Total (95% CI)4118175100.0%0.61 (0.49-0.76)Heterogeneity: Tau2=0.02; Chi2=5.58, df=4 (p=0.23); I2=28%Test for overall effect: Z=4.43 (p<0.00001)

Citation Format: Chun Philip Yeung, Siu Tim Cheung, Kwok Chai Kelvin Ng, Bo San Paul Lai, Ching Ning Charing Chong. Statin use associated with reduced risk of hepatocellular carcinoma recurrence following liver resection: A systematic review and meta-analysis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2345.
Abstract 2347: Incidence and mortality rate of liver and intrahepatic bile duct cancer in the United States, 2000-2016
Authors: Koko Michael Rutalira , Mutamba Glodi
Published: September 2020

Introduction: Liver and intrahepatic bile duct cancer is highly deadly. Incidence and mortality rates of Liver and intrahepatic bile duct cancer have increased significantly from 2000-2016 in the United States. The purpose of this study is to investigate the incidence and mortality rates of this cancer according to sex and race.

Methods: We used the Surveillance, Epidemiology, and End Results (SEER) 18 data to calculate age-adjusted incidence and mortality rates from 2000-2016. Annual Percent Change (APC) was obtained for incidence and mortality rates by sex and race.

Results: Between 2000 and 2016, the incidence rate increased significantly for both male and female from 8.9 to 13.0 and 3.3 to 4.7 per 100,000 persons, respectively. Additionally, the highest incidence rates have been reported among Hispanic, non-Hispanic American Indian/Alaska Native and non-Hispanic Asian or pacific Islander with 13.5, 14.6 and 11.6 per 100,000 persons, respectively. Overall mortality rate has increased specifically among male from 5.1 to 10.9 per 100,000 persons during the same period. Hispanic, non-Hispanic American Indian/Alaska Native and non-Hispanic Asian or Pacific Islander still have the highest mortality rate compared to other race/ethnicity with 11.3, 12.9 and 9.4 per 100,000 persons), respectively. From 2015 to 2016, incidence and mortality rates have decreased among all races and gender.

Conclusions: Hispanic, non-Hispanic American-Indian/Alaska native and non-Hispanic Asian or pacific Islander are at greater risk of developing and dying from this cancer. Additionally, males are more likely to acquire and die from this cancer. Prevention programs should focus on these populations in order to reduce the burden of this cancer. Further studies are necessary to examine the alarming increase in overall incidence and mortality rates of this cancer.

Citation Format: Michael Rutalira Koko, Glodi Mutamba. Incidence and mortality rate of liver and intrahepatic bile duct cancer in the United States, 2000-2016 [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2347.
Abstract 240: Inhibition of glycogen degradation sensitizes glioblastoma cells to ionizing radiation
Authors: Hendriks Anne M. , Zois Christos E. , Haider Syed B. , Pires Elisabete , Kalamida Dimitra , Lagerholm Christoffer , Morris John , McCullagh James S. , Fehrmann Rudolf S. , den Dunnen Wilfred F. , Harris Adrian L. , Jalving Mathilde
Published: September 2020

Glycogen storage has been demonstrated in multiple cancer cell lines and is upregulated in response to hypoxia both in vitro and in vivo. Evidence supports a potential role of the “glycogen shunt”, that describes the cycling of glucose through glycogen to subsequently fuel glycolysis, in brain tumors. Glioblastoma (GBM) is the most common and most deadly central nervous system tumor, and is characterized by hypoxia and metabolic reprogramming. We found that inhibition of glycogen degradation by downregulation of glycogen phosphorylase liver isoform (PYGL) in GBM cells led to a decrease in clonogenic growth and sensitization to ionizing radiation (IR) doses of 10-12 Gy. Control cells also showed growth impairment but resumed growth 10 days after IR exposure, while PYGL knockdown cells did not. To understand these different responses to high dose IR, we studied the intra-cellular effects of IR in control and PYGL knockdown GBM cells. Five days after IR exposure of PYGL knockdown cells, mitotic catastrophe and a senescence-like morphology were seen. Dysregulation of autophagy, accumulation of elongated and fragmented mitochondria, decrease in oxygen consumption rate and extracellular acidification rate, and increased pAMPK and pACC expression were shown in irradiated PYGL knockdown cells compared to control cells, suggesting metabolic stress. In human GBMs, PYGL mRNA expression was higher than in normal brain tissue and low PYGL mRNA expression was correlated with improved survival rates (hazard ratio 1.32, 95% confidence interval 1.08 - 1.61, P = 0.006). Immunohistochemical PYGL expression was also increased in human GBM versus normal brain tissue. In conclusion, inhibition of glycogen degradation sensitizes GBM cells to high dose IR, the cornerstone of GBM treatment, and may provide a way forward to improve treatment of primary GBMs.

Citation Format: Anne M. Hendriks, Christos E. Zois, Syed B. Haider, Elisabete Pires, Dimitra Kalamida, Christoffer Lagerholm, John Morris, James S. McCullagh, Rudolf S. Fehrmann, Wilfred F. den Dunnen, Adrian L. Harris, Mathilde Jalving. Inhibition of glycogen degradation sensitizes glioblastoma cells to ionizing radiation [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 240.
Abstract 2402: Apoptosis induced by non-thermal atmospheric pressure bio-compatible soft jet plasma on brain tumor through p38/MAPK signaling pathways
Authors: Han Ihn
Published: September 2020

Brain tumor is one of the most lethal forms of cancer to human beings, among which Glioblastoma astrocytoma is the most familiar and quickly progressing type of astrocytic brain tumor in adults with poor prognosis are still a therapeutic challenge. Current therapy options at diagnosis are multimodal and include surgical resection, radiation, and chemotherapy but difficulty to failure of drug delivery through the blood-brain barrier, tumor response to chemotherapy and intrinsic resistance of tumor cells. Therefore new therapy is needed that can afford selective targeting of proliferating cells to promote current therapies. Non-thermal plasma can be considered as a potential cancer therapy for the alteration of biological and clinical applications by generating reactive oxygen species (ROS) and reactive nitrogen species (RNS). In our study, we designate the effects of a NBP-J device to U87MG cell line. Previous research work established that jet plasma could induce apoptosis of various cancer cells, in particular indicative the selective cytotoxicity of cancer cells over normal cells. In this study, we assessed the effects of NBP-J on the U87MG brain cancer cell line including different effects and the physicochemical and biological correlation between the RONS cascade and p38/MAPK signaling pathway on the dose (time) dependent manner. The results of this study shows that the soft jet plasma significantly inhibits cell proliferation, inhibit survival pathway and induce apoptosis by p38/MAPK pathway in U87MG cells. The current results reveal that soft jet plasma has a powerful cytotoxic effect against brain cancer cells. Taken together, our results showed that NBP-J can become potential applications for effective and safe clinical trials for cancer therapy.

Keywords: Non-thermal atmospheric-pressure plasma, soft jet plasma, Human glioblastoma, Cancer therapy.

Acknowledgement

This work was supported by the Leading Foreign Research Institute Recruitment Program through the National Research Foundation of Korea (NRF- 2016K1A4A3914113) funded by the Ministry of Science, ICT, and Future Planning (MSIP) of the Korean Government for EH. Choi, I. Han. Also this work was supported by the Basic Science Research Program through the Ministry of Science ICT of Korea (2019R1H1A2101686) for I. Han.

Citation Format: Ihn Han. Apoptosis induced by non-thermal atmospheric pressure bio-compatible soft jet plasma on brain tumor through p38/MAPK signaling pathways [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2402.
Abstract 2410: Supraphysiological androgens induce ferroptotic cell death in prostate cancer cells
Authors: Kumar Rajendra , Boyapati Kavya , Thomas Naiju , Topiwala Deven , Kanacharoen Suthicha , Owoyemi Olutosin , Coffey Max , Carducci Michael , Markowski Mark C. , Antonarakis Emmanuel S. , Pardoll Drew , Denmeade Samuel , Kachhap Sushant K.
Published: September 2020

Castration resistant prostate cancer (CRPC) first manifests as a sustained rise in the androgen-responsive gene, PSA, consistent with reactivation of a functioning androgen receptor (AR) axis. This observation led to the development of “second-line” therapies aimed at further blocking androgen/AR signaling. Unfortunately, resistance to these agents can also develop quickly. Paradoxically, several studies have suggested that the growth of AR-positive human CRPC cell lines may be inhibited by supraphysiologic levels of testosterone (SupT). These studies suggested that the adaptive reliance on AR signaling by CRPC cells becomes a therapeutic liability that can be exploited through the administration of SupT, which we termed as bipolar androgen therapy (BAT). Understanding how BAT works at the molecular and cellular levels might help in rationally combining BAT with other agents to achieve increased efficacy and tumor responses. Our data indicates that SupT induces autophagy mediated degradation of ferritin in prostate cancer (PCa) cells. Degradation of ferritin results in increase in labile iron pool increasing lipid peroxidation. Our data further indicates that SupT distinctly induces ferroptosis, a nonapoptotic regulated form of cell death induced by the accumulation of labile iron. Ferroptosis is thought to have tumor suppressing capabilities by clearing tumor cells via immune system activation. BAT has been discussed as a potential therapy for prostate cancer, but further research is needed to understand its full potential. Future combination of BAT with existing immunotherapeutics including immune checkpoint blockade may prove beneficial for treatment of CRPC.

Citation Format: Rajendra Kumar, Kavya Boyapati, Naiju Thomas, Deven Topiwala, Suthicha Kanacharoen, Olutosin Owoyemi, Max Coffey, Michael Carducci, Mark C. Markowski, Emmanuel S. Antonarakis, Drew Pardoll, Samuel Denmeade, Sushant K. Kachhap. Supraphysiological androgens induce ferroptotic cell death in prostate cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2410.
Abstract 2461: Targeting eIF4A3 as a multifunctional translation initiation factor for breast cancer therapy
Authors: Ling Jun , Huang Allen , Ohno Ai , Chen Joe , Cavataio Maria
Published: September 2020

eIF4A3 is a unique eukaryotic translation initiation factor (eIF) with broad functions in the whole process of RNA metabolism, including mRNA splicing via exon junction complex (EJC), mRNA nuclear export, translation, and RNA surveillance through nonsense-mediated RNA decay (NMD). These post-transcriptional processes are highly involved in cancer development. However, the roles of eIF4A3 in cancers have not addressed. In this study, by analyzing a breast cancer patient database, we identified that the up-regulation of eIF4A3 is correlated with poor survival of triple negative breast cancer (TNBC) patients. Overexpression of eIF4A3, along with other EJC components (Magoh, Y14 and CASC3), were observed in multiple TNBC cell lines as compared to human normal cells and mouse normal tissues. Knockdown of eIF4A3 by RNAi was found to strongly inhibit TNBC cell proliferation in vitro; down-regulation of endogenous eIF4A3 also suppressed tumor growth in a mouse model. Dominant negative eIF4A3 and an ATP-binding defective mutants inhibited TNBC cell growth in the clonogenic assay. Furthermore, TNBC cells were found to be more sensitive to eIF4A3-specific inhibitors than luminal A and B subtypes of breast cancer cells. Thus, our study elucidates a strong cancer-promoting activity of eIF4A3 in breast cancer growth at cellular and animal model levels, warranting a new strategy to target eIF4A3 for breast cancer therapy, especially for TNBC as the most difficult subtype to treat.

Citation Format: Jun Ling, Allen Huang, Ai Ohno, Joe Chen, Maria Cavataio. Targeting eIF4A3 as a multifunctional translation initiation factor for breast cancer therapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2461.
Abstract 2467: Interaction of CYP3A5 with AR-HSP90 complex regulating AR activation in prostate cancer cells
Authors: Gorjala Priyatham , Goodman Oscar , Mitra Ranjana
Published: September 2020

Background: Previously we have demonstrated that CYP3A5 stimulates prostate cancer cell growth by promoting AR activation and downstream signaling. We further investigated the mechanism of CYP3A5's interaction with the AR-HSP90 complex known to regulate AR activation process. AR activation is tightly regulated and it involves several chaperons (HSP90, HSP70, HSP40, and HSP27), multiple cofactors and protein kinases. We hypothesize that stoichiometric CYP3A5 binding to the HSP90-AR complex can induce conformational changes in the chaperone complex, promoting AR nuclear migration and activation.

Method: We used co-immunoprecipitation to identify the interaction of CYP3A5 with the components of the AR-HSP90 complex. Immunofluorescence studies were performed to identify the components of the AR-HSP90 complex that co-localize with CYP3A5. FRET analysis was used to confirm the interactions of CYP3A5 with the AR-HSP90 complex.

Results: We performed co-immunoprecipitation (co-IP) experiments with extracts from flag-tagged CYP3A5 transfected LNCaP cells. Our data demonstrates that CYP3A5 co-immunoprecipitates with HSP90, AR, HSP70 and HSP40, all known components of the HSP90-AR complex. We further confirmed using reverse immunoprecipitation with AR/ HSP90 antibodies and agarose A/G beads that CYP3A5 is a part of the HSP90-AR mature complex. Empty vector transfected cell extract and IgG were used as controls for pull down and reverse IP respectively. Immunofluorescence studies demonstrated that HSP70 and HSP40 co-localized with CYP3A5 in the cytoplasm. Interestingly, the MDAPCa2b and LNCaP cells show co-localization of CYP3A5 and AR in the nucleus, in contrast to non-transformed RWPE1 cells where CYP3A5 remains in the cytoplasm. The nuclear presence of CYP3A5 is more pronounced in MDAPCa2b cells carrying the wild type CYP3A5. The FRET analysis further confirmed direct interaction between CYP3A5 and HSP40 a chaperon in the AR-HSP90 complex.

Conclusions: Our data confirms that CYP3A5 directly interacts with the AR-HSP90 complex regulating AR activation process. Further understanding of this interaction can help establish CYP3A5-AR axis as a novel therapeutic target.

Citation Format: Priyatham Gorjala, Oscar Goodman Jr., Ranjana Mitra. Interaction of CYP3A5 with AR-HSP90 complex regulating AR activation in prostate cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2467.
Abstract 2474: Potent small molecule TEAD inhibitors targeting the Hippo pathway exhibit antiproliferation in vitro and anti-tumor effect in vivo
Authors: Amidon Ben , Syed Sakeena , Cavanaugh Jill , Frosch Hyejin , Natarajan Prabitha , Ecsedy Jeffrey , McGovern Karen , Castro Alfredo C.
Published: September 2020

The Hippo signaling cascade is an important pathway for cancer biogenesis and tumor maintenance. The Hippo pathway is heavily mutated across many cancer indications through loss of function mutations in genes such as NF2. These pro-tumor mutations lead to the constitutive activation of the downstream transcriptional coactivators YAP and TAZ that drive the expression of many pro-survival and proliferation genes through the essential interaction with a TEAD protein family member. In addition, this unrestrained transcriptional program drives enhanced immune suppression in the tumor microenvironment. To target this oncogenic pathway we identified novel small molecule inhibitors that selectively bind to TEAD and disrupt their interaction with YAP and TAZ thereby downregulating YAP- and TAZ-dependent transcription. These inhibitors prevent TEAD palmitoylation which is essential for the interaction between YAP and TEAD. Furthermore, they inhibit in vitro proliferation of YAP-dependent (i.e. Hippo pathway-deficient cancer cell lines), but not Hippo pathway wild type cancer cell lines. An attraction of targeting this pathway is that many differentiated cells and tissues do not rely on the Hippo pathway, but notable exceptions have been demonstrated with tissue specific mouse knockouts in the bile duct and kidney podocytes. Importantly, our compounds did not affect survival of a differentiated mouse podocyte cell line or compromise mouse kidney histology. Subsequent experiments in vivo demonstrate these inhibitors downregulate YAP-dependent genes in human tumor xenografts after oral dosing. In addition, these inhibitors exhibit single agent tumor growth inhibition of human tumor xenografts in mice at well tolerated oral doses. These data demonstrate the potential for targeting this critical pathway in cancers with small molecules. Additional studies to identify tumor types that are Hippo pathway-driven and dependent on TEAD function are in progress.

Citation Format: Ben Amidon, Sakeena Syed, Jill Cavanaugh, Hyejin Frosch, Prabitha Natarajan, Jeffrey Ecsedy, Karen McGovern, Alfredo C. Castro. Potent small molecule TEAD inhibitors targeting the Hippo pathway exhibit antiproliferation in vitro and anti-tumor effect in vivo [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2474.
Abstract 2497: Topological data analysis revealed complexity behind ER positive breast cancer subtypes in TCGA transcriptome data
Authors: Koyama Takahiko , Saenz Aldo G. , Parida Laxmi
Published: September 2020

Introduction:

Breast cancer subtyping is still not fully agreed among oncologists. Today, many guidelines separate patients into four subtypes, luminal A, luminal B, Her2 positive, and triple negative breast cancers. However, distinction between luminal A and luminal B cases is not clear cut although many use Ki67 index. As more treatment options including immune checkpoint blockade are available, it is imperative to define strata and to find optimum treatments for each stratum.

Method:

TCGA BRCA transcriptome data (BRCA-UQ) is used, which has a sample size of 1089. We employed topological data analysis (TDA) which can handle high dimensional nature of transcriptome data in elegant manner. For characterization of the clusters, we used clinical data such as metastasis status, tumor sizes, age of diagnosis, survival, and pathological data and molecular data including PIK3CA, TP53, GATA3 mutations, MYC amplification, 8p23.2 deletion, 16q24.3 deletion, and tumor mutation burden (TMB). In addition, state of tumor microenvironment is assessed using CD45 expression for tumor infiltrating leukocytes (TIL) and HOXD12 expression for cancer associated fibroblast (CAF).

Results:

TDA cleanly clustered breast cancer transcriptome data separating ER+, Her2+ and triple negative cases. However, among ER+ cases, boundary between Luminal A and Luminal B is vague except for ER+/Her2+ cases. In ER+ cases, there are four clusters with poor prognosis. One has PIK3CA mutation, high MYC, and low TIL. Another cluster is characterized by GATA3 mutations. The other has low TIL and PIK3CA mutations. Also, high CAF cluster coincides with poor prognosis ones.

Conclusions:

The study revealed complexity behind ER+ breast cancer stratification from molecular and clinical perspectives. A TDA based method will likely to classify a patient into a right stratum better than conventional methods. Identified clusters with high TIL and high TMB which are good candidates for immune checkpoint blockade. Further study on identification of optimal treatments for each stratum will be warranted.

Citation Format: Takahiko Koyama, Aldo G. Saenz, Laxmi Parida. Topological data analysis revealed complexity behind ER positive breast cancer subtypes in TCGA transcriptome data [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2497.
Abstract 252: How the gatekeeper helix of the thioesterase domain of fatty acid synthase controls substrate recognition and initiation of cleavage of the proper substrate
Authors: Fromm Joseph , Teriete Peter , Cieplak Piotr , Misson Laetitia , Burkart Michael , Cosford Nicholas
Published: September 2020

Since the initiation of multiple clinical trials of two chemically distinct small molecule fatty acid synthase (FASN) inhibitors for the treatment of Non-Alcoholic SteatoHepatitis (NASH), breast cancer, colon cancer, and non-small cell lung cancer, FASN has garnered a resurgent interest as a therapeutic target. However, little effort has been made to better understand this target to improve upon previous drug discovery efforts. Using both computational modelling and enzymatic analysis, we have identified specific residues from the thioesterase (TE) domain of FASN that are crucial for the enzymatic function of FASN-TE while independent of the catalytic triad. Introduction of point mutations to these residues hinder the protein's ability to cleave the proper substrate of FASN, effectively disrupting the de novo synthesis of fatty acids. We propose a mechanism-of-action by which FASN-TE measures and initiates the cleavage of the proper substrate of FASN and have shown that interfering with the residues involved in this mechanism cripples the activity of the protein. Building upon these findings regarding the substrate selection mechanism of FASN-TE will allow for the enhanced design and development of the next generation of potent and selective inhibitors of FASN for the treatment of a variety of cancer types and other metabolic diseases.

Citation Format: Joseph Fromm, Peter Teriete, Piotr Cieplak, Laetitia Misson, Michael Burkart, Nicholas Cosford. How the gatekeeper helix of the thioesterase domain of fatty acid synthase controls substrate recognition and initiation of cleavage of the proper substrate [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 252.
Abstract 2539: Mutant TP53 action in lung dictated by the cell microenvironment
Authors: Yin Christopher , Gao Li , Villalona-Calero Miguel A. , Duan Wenrui
Published: September 2020

TP53 is a well-known tumor suppressor protein involved in many anti-tumor processes such as apoptosis, DNA repair, and cell cycle arrest. Codon 273 is one of the most frequently mutated sites among all p53 mutant types in human lung cancer. Recently, it has been discovered that TP53 and its mutants influence the expression of cancer-relevant microRNAs and vice-versa. Although many experiments have been performed on cell lines, far fewer experiments were geared toward how TP53 mutations affect miRNA expression in complex, in vivo settings. In addition to this, there are competing viewpoints on how TP53-273H could interact with wild type TP53. To investigate the mechanism involved in the oncogenic potential of mutant TP53 protein in vivo, TP53+/+/TG273H transgenic mice were bred into p53 null background. At the age of 5-6 weeks, TP53−/−; TP53−/-/TG273H; TP53+/+; and TP53+/+/TG273H mice were used to evaluate the expression of micro RNAs (miRNAs) as well as TP53 downstream genes. Based on the nanoString miRNA analysis, TP53−/- 273H mice exhibits decreased expression of miR-205, 206, and 1274a when compared against TP53−/− mice. However, TP53+/+ 273H mice resulted in increased expression of these miRNAs when compared against wild-type mice without mutant TP53 (TP53+/+). When comparing p53−/− knockout mice with wild-type mice, miRNAs 205, 206, and 1274a all increased at least 3-fold compared to wild-type. TP53−/− 273H mice exhibited widespread decrease of miRNA expression levels when compared against TP53+/+ 273H mice. The results suggest that the biological function of the TP53-273H is dictated by the cell microenvironment, especially the presence/absence of the wild type TP53 through dominant-negative effects in lung tissues. Mutant TP53-273H exerts oncogenic/tumor suppressor effects through specific miRNAs and their target gene networks.

Citation Format: Christopher Yin, Li Gao, Miguel A. Villalona-Calero, Wenrui Duan. Mutant TP53 action in lung dictated by the cell microenvironment [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2539.
Abstract 2569: Uncovering metabolic bottlenecks in KEAP1 mutant lung cancer
Authors: LeBoeuf Sarah E. , Wu Warren , Karakousi Triantafyllia , Wild Robert , Papagiannakopoulos Thales
Published: September 2020

During tumorigenesis cancer cells continuously encountermetabolic bottlenecks as a result of accelerated growth, overall increased metabolic demand and increased oxidative stress due to the formation of reactive oxygen species. We use a combination of genetically-engineered mouse models, an accelerated CRISPR/Cas9-based experimentalplatformand biochemical approaches to identify metabolic liabilities that can be exploited using novel targeted therapies.Lung cancer, the leading cause of cancer-related deaths worldwide,is the most common cancer type to acquire mutually exclusive gain-of-function mutations in the anti-oxidant transcription factor NRF2or loss-of-function mutations in its negative regulator KEAP1. Loss of Keap1activates Nrf2, increases antioxidant production and dramatically accelerates KRAS-driven lung cancer. Our data are in line with mounting evidence demonstrating that antioxidants promote cancer progression. Our studies focus on elucidating the mechanisms underlying novel metabolic dependencies in KEAP1mutant tumors and explore the therapeutic potential of targeting metabolism in highly relevant pre-clinical mouse and human lung cancer models. We observe that the ability of KEAP1mutant tumors to divert their metabolism towards antioxidant production comes with a cost, generating multiple metabolic vulnerabilities, including a dependency on glutamine metabolism that can be therapeutically exploited using DRP-104, a novel broad acting glutamine antagonist which inhibits all glutamine-consuming reactions. In addition, we have extended our findings to demonstrate that cancers with high antioxidant capacity exhibit a general dependency on exogenous non-essential amino acids (NEAAs) that is driven by the Nrf2-dependent secretion of glutamate through system xc-(XCT), which limits intracellular glutamate pools that are required for NEAA synthesis. This dependency can be therapeutically targeted by dietary restriction or enzymatic depletion of individual NEAAs. Importantly, limiting endogenous glutamate levels by glutaminase inhibition can sensitize tumors without alterations in Keap1/Nrf2 pathway to dietary restriction of NEAAs. Our findings identify a metabolic strategy to therapeutically target cancers with genetic or pharmacologic activation of the Nrf2 antioxidant response pathway by restricting exogenous sources of NEAAs.

Citation Format: Sarah E. LeBoeuf, Warren Wu, Triantafyllia Karakousi, Robert Wild, Thales Papagiannakopoulos. Uncovering metabolic bottlenecks in KEAP1 mutant lung cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2569.