bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P4-02-10: Pfkfb3 inhibition significantly decreases endocrine-resistant breast cancer growth and induces necroptotic cell death
Authors: Jones Brandon C , Sengupta Surojeet , Sevigny Catherine M , Jin Lu , Pohlmann Paula R , Shajahan-Haq Ayesha , Clarke Robert
Published: March 2022

Estrogen receptor-positive breast cancers constitute the majority of newly diagnosed breast cancers in patients. While commonly treated with various approved endocrine therapy strategies, these cancers can unfortunately also develop resistance to such therapies, indicating a need to explore other avenues of treatment in endocrine-resistant breast cancer. Rapidly dividing cancer cells are known to have an increased energy demand and altered reprogramming of glucose metabolism, leading to a subsequent heavy reliance on glycolysis. A critical rate-limiting step in the glycolysis pathway, the conversion of fructose-6-phosphate to fructose-1,6-bisphosphate, is greatly influenced by an allosteric activator produced by the enzyme PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3), thus making PFKFB3 an essential regulator of glycolytic flux. Furthermore, PFKFB3 has been shown to be overexpressed in many human cancers, including breast cancer, as well as be upregulated by estrogen. Through the following work, we show that endocrine-resistant BC cell growth is strongly reduced by PFKFB3 targeting using different commercial PFKFB3 inhibitors, which can be further exacerbated by combination treatment with well-established endocrine therapies like fulvestrant in some cell lines. Moreover, we relatedly show that PFKFB3 inhibition reduced tumor size in mouse xenografts of endocrine-resistant BC cells. Endocrine-resistant cells also presented higher basal glucose uptake and greater reduction of glucose uptake in response to PFKFB3 inhibition than endocrine-sensitive cells, indicating that endocrine-resistant breast cancer cells may have a more significant reliance on glycolysis and PFKFB3 activity for their metabolic needs than sensitive cells. Additionally, PFKFB3 inhibition produced an increase in phosphorylation of the necroptotic markers RIPK1 (receptor-interacting kinase 1), and MLKL (Mixed Lineage Kinase Domain Like Pseudokinase), and pre-treatment with the necroptosis inhibitor necrostatin-1 was able to partially rescue cell death caused by PFKFB3 targeting, suggesting that necroptosis is a primary method of cell death induced by PFKFB3 inhibition. In summation, this study showcases the glycolytic enzyme PFKFB3 as an encouraging therapeutic target in endocrine-resistant breast cancer, and warrants further review as a monotherapy or in combination with current endocrine therapies.

Citation Format: Brandon C Jones, Surojeet Sengupta, Catherine M Sevigny, Lu Jin, Paula R Pohlmann, Ayesha Shajahan-Haq, Robert Clarke. Pfkfb3 inhibition significantly decreases endocrine-resistant breast cancer growth and induces necroptotic cell death [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-02-10.
Abstract P4-05-08: 18F-FDG uptake of visceral adipose tissue on preoperative PET/CT as a predictive marker for breast cancer recurrence
Authors: Paik Pill Sun , Yoon Chang Ik , Kim Hyun Jeong , Cha Yoon Jin , Kim Dooreh , Bae Soong June , Ahn Sung Gwe , Jeong Joon , Park Woo-Chan , Ryu Young Hoon , Jeon Tae Joo , Kim Chai Won
Published: March 2022

Background: Glucose utilisation of visceral adipose tissue (VAT) reflects inflammatory activities, which promote tumour cell growth and carcinogenesis. The impact of the VAT-inflammatory response on survival outcomes in breast cancer is not known. We investigated survival outcomes in breast cancer patients based on the standardised uptake value (SUV) in VAT using 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET/CT). Methods: We retrospectively enrolled breast cancer patients (stages I-III) treated at the Gangnam Severance Hospital between 2007 and 2010. Metabolic activities of VAT and tumour were calculated based on FDG uptake, and the clinical characteristics and survival outcomes were comparatively analysed in the high-SUV and low-SUV groups. Results: The SUV of VAT (SUV-VAT) was obtained for 148 patients. With no significant intergroup differences in clinical characteristics, high SUV-VAT was associated with poor recurrence-free survival (RFS; hazard ratio [HR] 2.754, 95% confidence interval [CI] 1.090-6.958, P=0.032) and distant metastasis-free survival (DMFS; HR 3.500, 95% CI 1.224-10.01, P=0.019). Multivariate analysis showed that high SUV-VAT was a significant clinical factor for both poor RFS and DMFS (P=0.023 and P=0.039, respectively). SUV-tumour was significantly associated with RFS (P=0.0388), but not DMFS (P=0.0718). Conclusion: The glucose metabolism of VAT, assessed by 18F-FDG PET/CT, is a prognostic biomarker of adverse outcomes and of primary tumour metabolism in breast cancer. The glucose utilisation of VAT may reflect the degree of inflammation associated with breast cancer recurrence and metastasis. Keywords: breast cancer, visceral adipose tissue, inflammation, 18F-FDG, PET/CT, glycolysis, metastasis, recurrence

Citation Format: Pill Sun Paik, Chang Ik Yoon, Hyun Jeong Kim, Yoon Jin Cha, Dooreh Kim, Soong June Bae, Sung Gwe Ahn, Joon Jeong, Woo-Chan Park, Young Hoon Ryu, Tae Joo Jeon, Chai Won Kim. 18F-FDG uptake of visceral adipose tissue on preoperative PET/CT as a predictive marker for breast cancer recurrence [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-05-08.
Abstract P4-05-11: The clinical importance of the new category, ER low-positive in the ER expression HER2 negative early breast cancer
Authors: Wakako Tajiri
Published: March 2022

In the latest updated ASCO/CAP guideline, a new category, ER low-positive was introduced, which are determined by the 1% to10 % of positive staining of the tumor nuclei for ER. However clinical meaning of ER-low positive was unclear compared with ER- positive or ER-negative in HER2 negative breast cancer.  In order to clarify the clinicopathological features of ER-low positive breast cancer, we compared the clinicopathological factors and prognosis among three groups, ER negative, ER-low positive, and ER-positive groups in the patients with HER2 negative breast cancer.  A total of 1,882 patients with HER2 negative breast cancer who underwent surgery between 2001 and 2018 were included in this study. We divided them into three groups, ER negative (<1%), ER-low positive (1-10%), and ER positive (10%≦). The relationships between ER status and clinicopathological characteristics and prognosis were evaluated. According to ER status, 553(29.4%), 183(9.7%), 1146(60.9%) patients were divided into ER negative, low-positive and positive groups. Compared with ER-positive group, patients in the ER negative and low-positive groups were of higher age (p<.0001), and those tumors were significantly associated with the larger tumor size (p<.0001), higher histological grade (p<.0001) and more administration of adjuvant chemotherapy (p<0.0001). Most of the patients with ER low-positive and ER-positive tumors had received the adjuvant endocrine therapy. The prognosis of the patients with ER low-positive tumors were poorer than that of those with ER positive tumors in terms of relapse-free survival (RFS p<.0001), distant metastasis-free survival (DMFS p<.0001) and overall survival (OS p<.0001).In the node negative patients, the prognosis of the ER-negative and ER low-positive groups were equivalent, and poorer than that of the ER-positive group (RFS; p=0.0068, DMFS; p=0.0013, OS: p=0.0032). In addition, in pStage III patients, prognosis of the ER-negative and ER low-positive groups were also equivalent, and poorer than that of the ER-positive group (RFS; p=0.008, DMFS; p=0.0021, OS: p=0.0010).  This study showed that the prognosis of the patients with ER negative and ER low-positive were similar, and poorer than that of those with ER positive tumors. Therefore, the new category of ER-low positive is clinically especially important in order to determine the appropriate adjuvant therapy.

Citation Format: Tajiri Wakako. The clinical importance of the new category, ER low-positive in the ER expression HER2 negative early breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-05-11.
Abstract P4-07-20: Survival analysis using the entire transcriptome to pinpoint biomarkers with the highest prognostic power
Authors: Gyorffy Balazs
Published: March 2022

Introduction. Extensive research is directed to uncover new biomarkers capable to stratify breast cancer patients into clinically relevant cohorts. However, the overall performance ranking of such marker candidates compared to other genes is virtually absent. Here, we present the ranking of all survival related genes in chemotherapy treated basal and estrogen positive/HER2 negative breast cancer. Methods. We searched the GEO repository to uncover transcriptomic datasets with available follow-up and clinical data. After quality control and normalization, samples entered an integrated database. Molecular subtypes were designated using gene expression data. Survival analysis was performed using Cox proportional hazards regression. False discovery rate was computed to combat multiple hypothesis testing. Kaplan-Meier plots were drawn to visualize the best performing genes. Results. The entire database includes 7,830 unique samples from 55 independent datasets. Of those with available relapse-free survival time, 3,382 samples were estrogen receptor-positive and 696 were basal. In chemotherapy treated ER positive/ERBB2 negative patients the significant prognostic biomarker genes achieved hazard rates between 1.76 and 3.33 with a p value below 5.8E-04. The significant prognostic genes in adjuvant chemotherapy treated basal breast cancer samples reached hazard rates between 1.88 and 3.61 with a p value below 7.2E-04. Finally, an online accessible platform utilizing the entire database was set up enabling the validation of future biomarker candidates. Conclusions. A reference ranking for all genes in two chemotherapy treated breast cancer cohorts is presented. The results help to neglect those with unlikely clinical significance and to focus future research on the most promising candidates.

Citation Format: Balazs Gyorffy. Survival analysis using the entire transcriptome to pinpoint biomarkers with the highest prognostic power [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-07-20.
Abstract P4-07-21: High Tinagl1 expression is a marker of good prognosis in breast cancer
Authors: Kato Akiko , Kondo Naoto , Wanifuchi-Endo Yumi , Fujita Takashi , Asano Tomoko , Hisada Tomoka , Uemoto Yasuaki , Terada Mitsuo , Toyama Tatsuya
Published: March 2022

Background: Tubulointerstitial nephritis antigen-like 1 (Tinagl1) is reported to inhibit integrin and epidermal growth factor receptor (EGFR) and suppress tumor metastasis and growth in breast cancer. We investigated the association between Tinagl1 mRNA expression and clinicopathological factors and prognosis of breast cancer patients with long-term follow-up. The median follow-up period was 12.0 years. Materials and Methods: A total of 599 consecutive primary invasive breast cancer patients with available tissue from surgery in our hospital from 1992 to 2008 were included in this study. We examined the association between Tinagl1 mRNA expression and prognostic and clinicopathological factors using the TaqMan real-time PCR system. Statistical analysis was performed using the Kaplan-Meier method and the Cox proportional hazards model. Results: High expression of Tinagl1 mRNA was associated with significantly better disease-free survival (DFS) and overall survival (OS) than low expression (P=0.003 and P=0.01, respectively). In estrogen receptor (ER)-positive patients, DFS and OS were also better in the high Tinagl1 mRNA expression group (P=0.01 and P=0.03, respectively). However, there was no significant difference in DFS and OS in triple-negative breast cancer. Low Tinagl1 mRNA expression was positively associated with higher tumor grade. Multivariate analysis showed that low Tinagl1 mRNA expression, together with lymph node positivity, was an independent poor prognostic factor in DFS (HR 1.48; 95% CI 1.06-2.05, P=0.02). Conclusions: Our findings suggest that Tinagl1 has a role in the metastasis of breast cancer, and that its expression could be a promising candidate biomarker of prognosis in breast cancer patients.

Variablesn(%)DSF Univariate P valueDFS Multivariate P valueHR (95% CI)Nodal status590Negative349 (59.2)1Positive241 (40.8)<0.0001<0.00013.16(2.25-4.45)Tinagl1 mRNA expression599High300 (50.1)1Low299 (49.9)0.0030.021.48(1.06-2.05)

Citation Format: Akiko Kato, Naoto Kondo, Yumi Wanifuchi-Endo, Takashi Fujita, Tomoko Asano, Tomoka Hisada, Yasuaki Uemoto, Mitsuo Terada, Tatsuya Toyama. High Tinagl1 expression is a marker of good prognosis in breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-07-21.
Abstract P5-01-12: Physical and visual (social) isolation increases tumor aggressiveness in thermoneutral housing temperatures in murine mammary cancer models
Authors: Gaymon Darius O
Published: March 2022

Correct interpretation of tumor progression data , including the influence of host biology, in mouse models of breast cancer requires models and conditions that faithfully recapitulate human disease and human host status. Our previous attempts to investigate the effects of social isolation have proven inconclusive due to premature mortality in tumor-bearing animals. Those studies were completed in standard temperature (ST), which commonly is 70-72°F (21-22°C) for in vivo murine research based on laboratory animal care and use guidelines. Previous work from the Repasky lab (Kokulus, 2013) , which we have validated (Gaymon, 2020), demonstrates that ST housing results in chronic cold stress and immune suppression mediated by an increase in norepinrephrine (NE) levels, leading to increased tumor aggressiveness. Based on these findings, we investigated the effects of social isolation on BALB/cJ-4T1-luc and C57BL/6J/E0771 tumor progression and metastasis in thermoneutral housing conditions (84-85°F). Mice were first acclimatized to thermoneutral temperature and/or isolation for two weeks in cages that were unilaterally draped to provide physical and visual isolation. In BALB/cJ mice, 4T1-luc tumors were significantly larger in isolated mice compared to group-housed (GH) mice at day 18 (p<.0001). Statistically larger tumors were observed in isolated mice compared to GH mice through day 24 and final tumor masses were also significantly different (p=.004). Spleen masses were not statistically different. In C57BL/6J mice, E0771 tumors were significantly larger in isolates at Day 25 (p=.002). Final tumor masses were statistically (p=.002) different while no difference in spleen sizes were observed. Data on metastasis will be presented at the meeting. We hypothesized that social isolation may perturb immune function and next investigated the growth of 4T1-luc xenograft tumors in NSG mice. 4T1-luc/NSG tumor progression and metastasis data will also be presented at the meeting. We conclude that syngeneic breast tumor growth in immunocompetent BALB/cJ and C57BL/6J mice demonstrates that social isolation is a bona fide stress with sufficient influence to exacerbate breast cancer growth. These data are potentially clinically important due the known relationship of social support to survivorship outcomes in patients and the high-risk of depression and isolation in patients following breast cancer diagnosis. The data may provide additional insight into possible effects of COVID-19 isolation on breast cancer progression.

Citation Format: Darius O Gaymon. Physical and visual (social) isolation increases tumor aggressiveness in thermoneutral housing temperatures in murine mammary cancer models [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-01-12.
Abstract P5-06-01: Spatiotemporal gene expression analysis to identify novel tumor-neural crosstalk in TNBC brain metastases
Authors: Zhao Hong , Wang Xin , Sheng Jianting , Xu Shan , Gao Dongbing , Wong Stephen
Published: March 2022

Brain metastasis (BrM) occurs in 25-40% of TNBC metastatic patients while one-year survival remains a single digit. In contrast to other primary subtypes, TNBC usually spreads to the brain rapidly at early stages or even before the diagnosis of primary cancer. The shorter development time of BrM in TNBC and shorter survival time after BrM diagnosis may indicate a certain innate ability of TNBC cells in adapting to the brain. Recent seminal findings and our studies identified enriched and diverse expressions of neurotransmitters receptors, transporters, and key synthesis enzymes in brain metastatic TNBC cells. Since neurotransmitters, gliotransmitters, neurotrophic factors, and neural cytokines, called neuroactive substances (NSs), are the most abundant and key intercellular communication signals in the brain, we hypothesize that tumor cells leverage NSs to assimilate into the neural circuitry via communicating with various types of neural cells, including astrocytes, microglia/macrophages, oligodendrocytes and so on, to reinforce tumor-neural crosstalk in the development of BrM. By performing spatial transcriptomics experiments on mouse brain tissue sections bearing TNBC metastatic tumors, we found that brain regions, tumor lesions at different locations, and even tumor-surrounding areas were well separated into different clusters based on the spatial gene expressions and further predicted several novel paracrine and autocrine signalings between/within tumors and tumor-surrounding neural niches under different conditions, including tumor locations, tumor sizes, tumor center vs. edges, and stages of tumor development using the Cell-Cell Communication Explorer (CCCExplorer), a computational modeling tool for multi-cellular crosstalk and intracellular signaling developed by our lab. We conducted functional experiments to demonstrate that cell type-specific depletion of the secreted NSs deactivated the activities of corresponding receptor and downstream signaling using a cellular co-culture BrM system. The presented study indicates that the characterization and identification of spatiotemporal tumor-neural crosstalk in the BrM metastatic niche would open up a new vista for discovering novel therapeutics targets for devastating metastatic brain tumor.

Citation Format: Hong Zhao, Xin Wang, Jianting Sheng, Shan Xu, Dongbing Gao, Stephen Wong. Spatiotemporal gene expression analysis to identify novel tumor-neural crosstalk in TNBC brain metastases [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-06-01.
Abstract P5-06-15: Enabling therapeutic decisions for a breast cancer patient cohort using matched diseased and normal tissue in tumor organoids
Authors: Deems Liam , Shvartsman Dmitry , Ivanova Maria , Murphy Cheryl , Deems David
Published: March 2022

Pre-clinical cancer studies have been limited by the availability of patient cell models that accurately represent the diversity of patient populations. The utility of these studies is further impacted by the lack of normal tissue from the same patient. Furthermore, clinical evaluation of targeted therapeutics, like tarceva (erlotinib), would benefit from patient models expressing the target for robust characterization of drug response. Moreover, individual patient responses to the same anti-cancer compound cannot be assessed with high precision unless healthy controls are used from the same tumor donor. Here, we present six breast cancer patient-derived tumor models, accompanied by cancer (tumor)-associated fibroblasts (CAFs) and mesenchymal stem cells (MSCs). These paired and matched sample cohorts were studied in the context of spheroid models of the metastatic breast cancer niche. We show that these models allow an increased understanding of chemotherapy modulation by non-cancerous cells, which are present within the tumor but not targeted by the current chemotherapies or other anti-tumor approaches. We used a three-drug set to create a viability profile for each patient-associated tumor sample, which in turn can be used to aid clinical decision-making and increase the personalization of each treatment for an individual. Further, our approach shows different genomic and proteomic profiles for the co-culture models when compared between the tumor 3D and healthy controls.

Citation Format: Liam Deems, Dmitry Shvartsman, Maria Ivanova, Cheryl Murphy, David Deems. Enabling therapeutic decisions for a breast cancer patient cohort using matched diseased and normal tissue in tumor organoids [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-06-15.
Abstract P5-08-17: Small G protein RALA is a driver and potential therapeutic target in triple negative breast cancer
Authors: Richardson Dillon S. , Cole Matthew W. , Schafer Rachel E. , Spehar Jonathan M. , Steck Sarah A. , Das Manjusri , Lian Arthur W. , Ray Alo , Shakya Reena , Knoblaugh Sue E. , Timmers Cynthia D. , Sizemore Gina M. , Sizemore Steven T.
Published: March 2022

Triple Negative Breast Cancer (TNBC) is the leading cause of cancer mortality in women, mostly due to the lack of targeted treatment for this subtype of breast cancer (BC). RALA and RALB are small GTPases implicated in tumor proliferation, survival, and metastasis in a variety of cancers. However, little is known of their roles in breast cancer. Utilizing 3D spheroid invasion assays, we identified that knockout (KO) of RALA greatly reduced the invasion of MDA-MB-231 spheroids in basement membrane extract (BME). Conversely, RALB-KO significantly increased 3D invasion of MDA-MB-231 cells. We further investigated roles for RALA and RALB in TNBC with cell viability assays, transwell assays, and 3D growth assays. Results indicate that KO or depletion of RALA in TNBC cell lines MDA-MB-231 and MDA-MB-468 reduces cell viability and cell migration capabilities in vitro. On the contrary, loss of RALB increased cell migration and viability. Treating TNBC cells with a small molecule inhibitor of both RAL isoforms (BQU57) reduced cell growth in vitro as well as tumor growth and metastasis in vivo. Furthermore, RALA expression, but not RALB expression, was predictive of response to chemotherapy in TNBC patients and RAL inhibitor sensitized TNBC cells to paclitaxel. Combined, these results highlight the importance of the RALs, particularly RALA, as a therapeutic targets in TNBC.

Citation Format: Dillon S. Richardson, Matthew W. Cole, Rachel E. Schafer, Jonathan M. Spehar, Sarah A. Steck, Manjusri Das, Arthur W. Lian, Alo Ray, Reena Shakya, Sue E. Knoblaugh, Cynthia D. Timmers, Gina M. Sizemore, Steven T. Sizemore. Small G protein RALA is a driver and potential therapeutic target in triple negative breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-08-17.
Abstract P5-09-04: Regulation of mTOR signaling pathway by ING4 in breast cancer: A new target in cancer therapy
Authors: Shatnawi Aymen , Salisbury Travis
Published: March 2022

Background: Breast cancer (BC) is one of the most diagnosed tumors in the United States. The mechanism of the disease development and progression is complex and still not fully understood. Dysregulation or change in the expression of transcription factors has been proposed as a possible cause of the disease severity and poor prognosis. The INhibitor of Growth family member 4 (ING4) is a tumor suppressor that has been connected to various cancer-related processes. Therefore, this study aims to assess the global gene regulation of BC cells by ING4 and identify the associated cellular processes and molecular pathways related to BC following ING4 loss. Methods: siRNA Knockdown and CRISPR Knockout technologies were used to deplete ING4 expression. Real-time PCR, RNA-Seq and Chip-Seq were used to identify gene expression alteration and ING4 enrichment on target promoters. Ingenuity Pathways Analysis (IPA®) was used to highlight candidates, networks and cellular processes. Results: Protein and RNA levels of ING4 were significantly reduced in MDA-231 knockdown and MCF7 knockout cells using siRNA and CRISPR technologies. RNA-Seq of BC cells revealed a large set of differentially expressed genes were regulated following ING4 loss. Key genes and biological networks related to cellular processes, including amino acid metabolism, cancer, cellular development, cellular growth and proliferation, were identified using IPA®. Chip-seq also revealed that ING4 was recruited to genes important in BC development and progression. Several genes were validated using the RT-qPCR method, and results were consistent with RNA-seq and Chip-seq data. Importantly, MDA-231 cells grown in stripped serum have shown an upregulation of the ribosomal protein S6 phosphorylation (PS6), an mTOR target protein, following ING4 knockdown. Treatment of MDA-231 cells with rapamycin blocked the phosphorylation of S6 induced by ING4 loss. An increase in S6 phosphorylation was also observed in MCF7 knockout cells grown either in regular or serum-free growth medium. The phosphorylation was reversed by amino acid transporter inhibitors JPH and BCH. Conclusion: The present study shows that ING4 regulates many genes involved in biological pathways and networks related to BC development and progression. Importantly, ING4 is a novel regulator of the mTOR pathway. Collectively, regulation of ING4 gene expression and its downstream molecules provides a novel approach in cancer treatment.

Citation Format: Aymen Shatnawi, Travis Salisbury. Regulation of mTOR signaling pathway by ING4 in breast cancer: A new target in cancer therapy [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-09-04.
Abstract P5-10-08: Inflammation drives NOS2-Akt2 signaling in triple negative breast cancer
Authors: Chung Su , Montfort William
Published: March 2022

Triple negative breast cancer (TNBC) is aggressive and represents nearly 20% of all breast cancer cases. It is associated with poor prognosis due to its high metastatic capabilities and lack of targeted therapeutic treatments. To improve TNBC treatment, our goal is to better understand the molecular mechanisms that drive TNBC. Recently, inducible nitric oxide synthase (NOS2), a nitric oxide (NO) producing enzyme, has emerged as a key player in TNBC progression. High NOS2 expression in TNBC patient correlates with poor prognosis marked with increased metastasis and disease recurrence. NOS2 catalyzes L-arginine and dioxygen (O2) to produce citrulline and high, sustained levels of NO which has been shown to alter the activity of numerous signaling pathways. Depending on NO concentration and temporal and spatial levels, it has been reported to have both tumor-promoting and tumor-suppressing functions. However, the mechanism by which NOS2 drives TNBC is unclear. Akt is major oncogenic signaling pathway that regulates various aspects of tumor progression, and it has been shown that NO can activate Akt activity. The Akt family has three members, Akt1, Akt2, and Akt3, that seem to have similar and non-redundant functions in cancer. We have discovered for the first time that NOS2 specifically activates Akt2 in TNBC cells. Thus, we hypothesize that Akt2 plays a major role in NOS2 driven TNBC. Inflammatory cytokines (IFN-γ, IL-1β, and TNF-α) highly induce NOS2 in mouse 4T1 TNBC cells. We developed a real-time NO detection assay to measure NO levels in 4T1 cells and confirmed that these cells produce high levels of NO. By pharmacologically and genetically inhibiting NOS2 in the 4T1 cells, we found that NOS2 suppresses Akt2 phosphorylation but not the total levels. Furthermore, NOS2 had no effect on Akt1. The decrease in Akt2 phosphorylation was accompanied by inhibition of its downstream targets, GSK3β and FOXO3a, suggesting that NOS2 is regulating Akt2 activity. Conversely, treatment of 4T1 and human MDA-MD-231 TNBC cells with DETA/NO, a slow releasing NO donor, resulted in increased Akt2 phosphorylation with no effect on Akt1. Akt2 has been shown to promote migration, invasion, and metastasis. Using 4T1 cells, we are presently studying the function of NOS2-Akt2 signaling in TNBC. We have orthotopically injected 4T1 cells into immunocompetent mice and have found that 4T1 tumors express high levels of NOS2 as well as phosphorylated Akt2. We have knocked out NOS2 in the 4T1 cells to examine the effects of NOS2 inhibition in the mouse tumor model. Understanding the link between NOS2 and Akt2 will give further understanding on how NOS2 drives TNBC.

Citation Format: Su Chung, William Montfort. Inflammation drives NOS2-Akt2 signaling in triple negative breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-10-08.
Abstract P5-12-11: NSDHL knockdown decreases tightly cohesive tumorsphere formation and breast cancer stem cell population
Authors: Yoon So-Hyun , Kim Hoe Suk , Song Junhyuk , Kwon Sunkyu , Ryu Seungyeon , Baek Moonjou , Lee Sangeun , Lee Han-Byoel , Moon Hyeong-Gon , Noh Dong-Young , Jon Sangyong , Han Wonshik
Published: March 2022

Background: Cholesterol biosynthesis pathway is commonly elevated or dysregulated in cancer cells and high cholesterol levels are associated with cancer progression. NAD(P)-dependent steroid dehydrogenase-like (NSDHL), an enzyme involved in cholesterol biosynthesis, has been implicated in maintaining cancer stem cell, but its role in regulating breast cancer stem cells (BCSC) remains unclear. This study aimed to uncover the molecular mechanisms by which NSDHL regulates BCSC properties.Methods: The knockdown of NSDHL was induced by an introduction of siRNA/shRNA into MCF-7 cells. A non-adherent 3D culture method based on polymer-X coating was applied for growing tumorspheres under serum-free condition. 3’ quant mRNA-sequencing, qRT-PCR, western blot, immunofluorescent staining, ELISA, flow cytometry and ALDEFLUOR assay were carried out. Immunodeficient NSG mice were used for orthotropic xenograft tumor models by injecting tumorspheres.Result: NSDHL knockdown reduced a tightly cohesive sphere formation. RNA-seq data revealed a significant upregulation of 253 DEGs and downregulation of 364 DEGs with >2-fold change in NSDHL knockdown tumorsphere. The KEGG pathway analysis showed that DEGs were mainly enriched in TGF-β signaling pathway and cell cycle. NSDHL knockdown reduced the population of BCSCs with CD44+/CD24- phenotype and high ALDH activity as well as luminal progenitors with CD49f+/EpCAM+ phenotype along with decrease in TGF-β 1 and 3 and GLI1 in tumorsphere. In orthotropic xenograft tumor models, NSDHL knockdown strongly suppressed the tumor initiation and growth.Conclusions: Our findings reveal that NSDHL has an important role in maintaining BCSC population and tumor initiating capacity, suggesting NSDHL as an attractive therapeutic target to eliminate BCSCs, and thus preventing breast cancer initiation and progression.

Citation Format: So-Hyun Yoon, Hoe Suk Kim, Junhyuk Song, Sunkyu Kwon, Seungyeon Ryu, Moonjou Baek, Sangeun Lee, Han-Byoel Lee, Hyeong-Gon Moon, Dong-Young Noh, Sangyong Jon, Wonshik Han. NSDHL knockdown decreases tightly cohesive tumorsphere formation and breast cancer stem cell population [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-12-11.
Abstract P5-14-12: Health literacy among long term breast cancer survivors; exploring associated factors in a nation-wide sample
Authors: Vandraas Kathrine Flørenes , Reinertsen Kristin V , Kiserud Cecilie E , Bøhn Synne K , Lie Hanne C
Published: March 2022

Objectives: Low health literacy (HL) levels likely hamper health management and outcomes in cancer survivorship. Knowledge of associated factors is needed to identify at-risk sub-groups of survivors, but is currently scant. Here, we explore HL-levels and associated factors in a nation-wide sample of long-term BCSs. Methods: All BCSs aged 20-65 years, diagnosed with early-stage BC in 2011-2012, were identified through the Norwegian Cancer Registry and invited to participate in the Survivorship, Work and Sex (SWEET)-study. HL was measured using The European Health Literacy Survey Questionnaire-12 (HLS-EU-Q12) as a continuous and categorical variable. Associations between HL and sociodemographic-, physical- and mental health variables, including the most common LEs, were explored in uni- and multivariable linear regression models. Results: The final sample consisted of 1355 survivors (48%). Median age was 60 years and the majority of survivors had high socioeconomic status (SES). Median HL sum-score corresponded to an intermediate HL level (36.2, SD 5.4), while 19.3% reported marginal or inadequate HL. Education, income, age at diagnosis, the personality trait neuroticism and fear of cancer recurrence (FCR) remained significantly associated with HL in the multivariate model, explaining 12% of the variance in HL scores. Conclusion: Low levels of HL were prevalent, potentially affecting the provision of effective survivorship care and BCS’s self-management abilities. Identifying individuals at risk of low HL based on knowledge readily available to clinicians remains challenging and warrants further investigation. Implications for cancer survivors: Addressing survivors’ HL could be a key target for improving survivorship care.

Citation Format: Kathrine Flørenes Vandraas, Kristin V Reinertsen, Cecilie E Kiserud, Synne K Bøhn, Hanne C Lie. Health literacy among long term breast cancer survivors; exploring associated factors in a nation-wide sample [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-14-12.
Abstract P5-15-05: Influence of shared decision making with decision aids on decisional conflict and regret in breast cancer surgery: A randomized controlled trial
Authors: Tam Ka-Wai , Lin Shi-Qian
Published: March 2022

Background: Patients with breast cancer usually encounter difficulties in making treatment decisions. Shared decision making (SDM) is a way to minimize decisional conflicts of patients, and patients’ decision aids (PDAs) have been developed to support the implementation of SDM. However, the effect of PDAs in Asia is inconclusive. Therefore, we developed a randomized controlled trial to evaluate the effect of SDM assisted by PDAs on decisional conflict among patients with breast cancer in Taiwan. Methods: This is a two-group, outcome assessor-blind, randomized, controlled trial. A total of 151 patients diagnosed with breast cancer were assigned to the PDAs group or the classic group. Demographic and clinical variables were analyzed to identify the variables influencing the treatment choice. The patients’ decision-making difficulties were evaluated using the 4-item SURE scale during the pre-operative hospitalization, and the decisional conflicts were assessed by the 5-item Decision Regret scale and Hospital Anxiety and Depression Scale (HADS) at one month postoperatively.Results: The choice of breast conservation therapy and mastectomy was not significantly different between the PDA and classic groups. Compared with the classic group, patients in PDA group had more depression after making decision (P = 0.029). No significant difference was noted in terms of the total scores in the SURE scale and Decision Regret scale between the groups.Conclusions: Our study revealed that PDA did not assist breast cancer patients in decision making. Clinicians should focus on SDM based on evidence-based medicine with care, and help patients taking their individual preference into consideration.

Citation Format: Ka-Wai Tam, Shi-Qian Lin. Influence of shared decision making with decision aids on decisional conflict and regret in breast cancer surgery: A randomized controlled trial [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-15-05.
Abstract PD2-02: Combination CDK4/6 inhibition and AR agonism suppresses the growth of CDK4/6 inhibitor resistant breast cancers
Authors: Freelander Allegra , Laven-Law Geraldine , Eshraghi Leila , Chia Kee Ming , Pickering Marie , Yong Aliza , Wilkinson Ashleigh , Alexandrou Sarah , Caldon C. Elizabeth , Hickey Theresa E , Tilley Wayne D , Lim Elgene
Published: March 2022

Resistance to standard-of care-therapies is a significant clinical challenge in estrogen receptor positive (ER+) breast cancer. Cyclin dependent kinase 4/6 inhibitors (CDK4/6i) in combination with endocrine therapies (ET) is the current standard-of-care for advanced metastatic ER+ breast cancer; however, resistance to this combination is considered inevitable, leading to disease progression. The androgen receptor (AR) is expressed in up to 90% of all ER+ breast cancers, and has been associated with better patient outcome. Compelling recent pre-clinical data demonstrates that selective androgen receptor AR modulators (SARMs) act to suppress ER-driven tumour growth of endocrine-sensitive and -resistant models of ER+ breast cancer. Furthermore, a recent clinical trial evaluating the efficacy of SARMs has shown clinical benefit in patients with ER+/AR+ metastatic breast cancer (NCT02463032). We hypothesise that the SARMS, either alone or in combination with a CDK4/6i, would be an effective treatment for tumours that are sensitive and resistant to ET and CDK4/6i. We report therapeutic efficacy of combination AR agonism + CDK4/6i in CDK4/6-naïve and -resistant pre-clinical models, including cell lines and patient derived xenograft (PDX) models. We demonstrate that combination SARM + CDK4/6i potently and durably inhibited in vitro and in vivo tumour growth. Additionally, we provide evidence that in vivo treatment with the CDK4/6i Palbociclib increased AR expression and signalling, highlighting an interaction of the two signalling pathways not previously described. In conclusion, our data provides a pre-clinical rationale for combination SARM + CDK4/6i in CDK4/6i resistant ER+ breast cancer.

Citation Format: Allegra Freelander, Geraldine Laven-Law, Leila Eshraghi, Kee Ming Chia, Marie Pickering, Aliza Yong, Ashleigh Wilkinson, Sarah Alexandrou, C. Elizabeth Caldon, Theresa E Hickey, Wayne D Tilley, Elgene Lim. Combination CDK4/6 inhibition and AR agonism suppresses the growth of CDK4/6 inhibitor resistant breast cancers [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr PD2-02.
Abstract PD7-07: Surgical management and contralateral breast cancer risk in women with a history of radiation therapy for Hodgkin lymphoma: Results from a population-based cohort
Authors: Eisenberg Elisheva R , Weiss Anna , Prakash Ipshita , Skamene Sonia , Basik Mark , Boileau Jean Francois , Ajjamada Lissa , Pollak Michael , Wong Stephanie M
Published: March 2022

Background: Women with a history of chest radiation for Hodgkin lymphoma are at an increased risk of developing breast cancer. Although mastectomy has historically been recommended for surgical treatment of breast cancers in women with prior radiation exposure, surgical management trends and contralateral breast cancer risk in this population remain undefined. Methods: We performed a population-based retrospective study using the Surveillance, Epidemiology, and End Results (SEER) database from 1990-2016. Our cohort included women who received radiation for Hodgkin lymphoma prior to 30 years old and were diagnosed with a subsequent breast cancer. We evaluated trends in local therapy including rates of breast conserving surgery (BCS) and mastectomy. In those undergoing unilateral surgery, the Kaplan-Meier method was used to estimate the 5- and 10-year cumulative incidence of contralateral breast cancer. Results: Our final cohort included 295 women with a median age of 22 years (range, 8-30 years) at Hodgkin lymphoma diagnosis, and 42 years (range, 22-65 years) at breast cancer diagnosis. Of these patients, 263 (89.2%) presented with unilateral breast cancer, while 32 (10.8%) presented with synchronous bilateral breast cancer. Overall, BCS was performed in 17.3% of patients and mastectomy was performed in 82.7%. In the 263 patients presenting with unilateral breast cancer, 50 (19.0%) underwent BCS and 213 (81.0%) underwent mastectomy. Subgroup analysis of mastectomy patients with surgical laterality information available demonstrated a 40.5% bilateral mastectomy rate. In the entire cohort, the 5-year incidence of contralateral breast cancer in women who underwent unilateral surgery was 9.4% (95% CI, 5.6-15.4), increasing to 20.2% (95% CI, 13.7-29.2) at 10-years of follow up. Hormone receptor status of the index breast cancer was not associated with significant differences in the incidence of contralateral breast cancer (p=0.13). Conclusions: Women with a history of prior chest radiation for Hodgkin Lymphoma with a diagnosis with breast cancer have a 10-year contralateral breast cancer risk of 20%. These findings support consideration of contralateral prophylactic mastectomy during surgical decision-making in this high-risk patient population.

Citation Format: Elisheva R Eisenberg, Anna Weiss, Ipshita Prakash, Sonia Skamene, Mark Basik, Jean Francois Boileau, Lissa Ajjamada, Michael Pollak, Stephanie M Wong. Surgical management and contralateral breast cancer risk in women with a history of radiation therapy for Hodgkin lymphoma: Results from a population-based cohort [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr PD7-07.
The Regulation of Germline Sex Determination in Drosophila by Sex lethal
Authors: Grmai Lydia , Pozmanter Caitlin , Van Doren Mark
Published: March 2022

Background: The establishment of male or female identity (sex determination) is essential for creating the anatomical, physiological, and behavioral differences between 2 sexes of the same species (sexual dimorphism). In many organisms, including mammals and Drosophila, sex is determined by inheritance of sex chromosomes, while in other animals, sex is determined by environmental factors. Arguably the most important consequence of sex determination is the production of healthy gametes necessary for reproduction: female oocytes and male spermatids. Summary: The generation of sperm and oocytes requires cooperation between 2 different cell types within the gonad: germ cells and somatic cells. Defects in sex determination in either the somatic gonad or germline lead to disorders of sexual development and infertility. In Drosophila, the gene Sex lethal (Sxl) is the key determinant of sex in both the soma and the germline. However, how Sxl controls sex determination is much more well understood in the soma than the germline. Key Mesage: This review will focus on Sxl in the germline, how it is activated specifically in female germ cells, and how it regulates germline sex determination and sexual development.
Is There Any Link between Cognitive Impairment and Gut Microbiota? A Systematic Review
Authors: Angoorani Pooneh , Ejtahed Hanieh-Sadat , Siadat Seyed Davar , Sharifi Farshad , Larijani Bagher
Published: March 2022

Background: Symptoms of cognitive impairments vary from mild without clinical manifestation to severe with advanced signs of dementia or Alzheimer’s disease (AD). Growing evidence in recent years has indicated the association between the brain and gut microbiota, which has been described as the “gut-brain axis.” This systematic review seeks to summarize the primary results from recent human and animal studies regarding the alteration of gut microbiota composition in cognitive disorders. Methods: A systematic literature search was conducted on PubMed, Scopus, and Web of Science databases up to August 2020. The full texts of the papers were analyzed to retrieve the relevant information. Results: Totally, 24 observational studies (14 animal and 13 human studies) were included. Most of the animal studies were performed on mouse models of AD. Human studies were conducted on patients with Parkinson’s disease (3 studies), AD (4 studies), poststroke cognitive impairment patients (1 study), and patients with mild to severe cognitive impairment without mention to the cause of disease (5 studies). More recent evidence suggests that throughout aging Firmicutes and Bifidobacteria decrease but Proteobacteria increases. Conclusions: The gut microbiota may alter brain function or trigger various psychiatric conditions through the gut-brain axis. Prospective studies are needed in order to explore the role of the gut microbiota in the etiology of dementia and to achieve clinical recommendations.
Early Life Stress, Hormones, and Neurodevelopmental Disorders
Authors: Makris Gerasimos , Eleftheriades Anna , Pervanidou Panagiota
Published: March 2022

Background: Early life stress (ELS) describes a broad spectrum of adverse and stressful prenatal events, namely, prenatal maternal stress (PMS), or early postnatal events, which can have detrimental long-term influences on the physiology, cognition, and behavior of an individual. There is abundant evidence indicating that ELS exerts its lasting effects on the physical and mental health of the individual, likely acting through a number of mediating mechanisms, including the disruption of developmental programming of the fetus. Neurodevelopmental disorders (NDDs), for example, attention deficit hyperactivity disorder (ADHD) and autism spectrum disorder (ASD), are a group of conditions that typically manifest during infancy, childhood, or adolescence and are characterized by developmental deficits in various domains. Summary: The scope of the current mini-review is to provide an up-to-date summary of the findings regarding the association of ELS and NDDs and the possible hormonal mechanisms through which PMS exerts its impact on neurodevelopment. We focus on the available evidence regarding children and adolescents diagnosed with ADHD or ASD. ELS exposure during developmental vulnerability windows may increase the risk for either subclinical neuropsychological alterations or clinical conditions, such as NDDs. In fact, a large body of evidence underlies the association of ELS exposure and increased risk for NDDs in the offspring. Key Messages: The majority of data suggest that ELS, including PMS, may be associated with ADHD and ASD in the offspring, although there is no consensus regarding the critical developmental periods. Carefully controlled prospective studies are needed to determine the possible causal processes and mechanisms underlying the association of ELS and NDDs.