bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5713: Reprogramming of serine metabolism during breast cancer progression
Authors: Li Albert Mao , Ducker Gregory S. , Li Yang , Seoane Jose A. , Xiao Yiren , Melemenidis Stavros , Zhou Yiren , Liu Ling , Vanharanta Sakari , Graves Edward E. , Rankin Erinn B. , Curtis Christina , Massague Joan , Rabinowitz Joshua D. , Thompson Craig B. , Ye Jiangbin
Published: September 2020

The causes and consequences of metabolic reprogramming during human breast cancer metastasis remain largely unknown. We performed metabolomics in the human triple-negative breast cancer cell line MDA-MB-231 (231-Parental) and its metastatic subclones that display tissue-tropism towards the brain (831-BrM), the bone (1833-BoM), and the lung (4175-LM). Using molecular and metabolic flux analysis, we uncovered that the mitochondrial serine and one-carbon (1C) unit pathway is upregulated in the metastatic subclones. Inhibition of the first rate-limiting enzyme of the pathway, serine hydroxymethyltransferase (SHMT2), potently suppresses proliferation of metastatic subclones in culture and impairs growth of lung metastatic subclones at both primary and metastatic sites in mice. In addition to breast cancer, a few other cancer types, such as adrenocortical carcinoma (ACC) and kidney chromophobe cell carcinoma (KICH), also display increased SHMT2 expression during disease progression. Moreover, we found that metastatic cells are exquisitely adapted to low serine conditions by synthesizing serine de novo from glucose to generate 1C units, and blocking serine catabolism abrogates metastatic growth under serine deprivation.

How do metastatic breast cancer cells acquire new metabolic properties such as de novo serine synthesis? We observed that 231-Parental cells grown in media lacking serine for up to ten passages exhibited metastatic cell-like metabolic properties. Subsequent RNA-sequencing revealed dramatic global gene expression changes during serine starvation in more primary tumor-like cells. Several pathways involved in metastasis such as TNFα signaling, mTOR1 signaling, and Ras signaling were modulated upon serine starvation. These data suggest that the availability of the nonessential amino acid serine may play a key regulatory role in breast cancer progression and metastasis, and highlight the potential for metabolite supplementation to block or halt these transformations.

Citation Format: Albert Mao Li, Gregory S. Ducker, Yang Li, Jose A. Seoane, Yiren Xiao, Stavros Melemenidis, Yiren Zhou, Ling Liu, Sakari Vanharanta, Edward E. Graves, Erinn B. Rankin, Christina Curtis, Joan Massague, Joshua D. Rabinowitz, Craig B. Thompson, Jiangbin Ye. Reprogramming of serine metabolism during breast cancer progression [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 5713.
Abstract 5725: Epigenetic and transcriptional profiling reveal endosomal protein sorting as a glioblastoma cancer stem cell dependency
Authors: Gimple Ryan C. , Zhang Guoxin , Rich Jeremy N.
Published: September 2020

Glioblastoma is a highly lethal primary intrinsic brain tumor for which current standard-of-care therapies are merely palliative. Functionally defined glioblastoma stem cells (GSCs) contribute to this poor prognosis by driving therapeutic resistance, maintenance of cellular heterogeneity, and disease recurrence. A deeper appreciation for the specific features that distinguish normal from neoplastic stem cells may inform selective therapeutic targeting. To further understand the molecular processes essential for maintaining cancer stem cell populations, we interrogated the active enhancer landscapes and transcriptional output of GSCs and normal neural stem cells (NSCs) through H3K27ac ChIP-seq and RNA-seq respectively. GSC-specific targets that held prognostic significance in patient datasets and which were essential in whole genome CRISPR screening data implicated endosomal protein sorting as a key feature in GSC biology. Targeting endosomal sorting impaired GSC viability and proliferation and contributed to apoptosis, while NSCs displayed a reduced cell proliferation deficit. Impaired endosomal protein sorting led to a reduction in the mRNA and protein levels of the stem and neuro-developmental transcription factors SOX2 and OLIG2 and reduced self-renewal capacity of GSCs, suggesting a role in the maintenance of stemness. Mechanistically, regulation of endosomal sorting may impact localization and stability of receptor tyrosine kinases and WNT signaling elements to control GSC maintenance. High expression of this endosomal sorting factor is associated with poor glioblastoma patient prognosis, suggesting its clinical importance. Thus, targeting the endosomal sorting pathway may represent a novel therapeutic approach for glioblastoma treatment.

Citation Format: Ryan C. Gimple, Guoxin Zhang, Jeremy N. Rich. Epigenetic and transcriptional profiling reveal endosomal protein sorting as a glioblastoma cancer stem cell dependency [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 5725.
Abstract 5763: Incidence and mortality predictors of ill-defined illness (non-diagnosed conditions) as a cause of death in cancer patients: A population-based study
Authors: Elfaituri Muhammed Khaled , Morsy Sara , Huy Nguyen Tien
Published: September 2020

Introduction:

This observational study characterised ill-defined illness (non-diagnosed conditions) mortality risk for multiple cancer sites, with respect to other factors related to the patient's condition and demographic information.

Method: Using the Surveillance, Epidemiology and End Results (SEER) database (1975-2016), complete data on 24378 patients were identified; their death was due to non-diagnosed conditions were identified. Overall survival (OS) was estimated using the Kaplan-Meier method. Cox, the proportional hazards model, were used to produce adjusted hazard ratios (HR). Trend analysis was conducted using joinpoint regression analysis with four join points, and annual percentage change (APC) was calculated.

Results: We found that 24378 cancer patients died due to unknown conditions. The mean age of the patients was 71.22, and most of them were female (50.6%). Most tumours were malignant 92.2% and grade II (n = 6650 (27.3%)). Most patients had surgery, only. White and married patients constituted most of the patients. The most common site associated with the ill-defined condition was GIT (n = 4578) followed by breast (n = 4003) followed by the male genital system (n = 3630). The most common pathology associated with ill-defined conditions was adenoma and adenocarcinoma (n = 9575) followed by ductal and lobular carcinoma (n = 3501) followed by squamous cell carcinoma (n = 2181). There were five trends observed for the mortality due to undiagnosed conditions. The rate of cases increased significantly from 1975 to 2008; then there was a decrease in cases from 2008 till 2015, but it was small and insignificant followed by a significant decrease in the number of cases [-59.4, (95% CI = -71.4; -42.5), p-value <0.001]. The overall average annual percent change was significant increase was 4.8% (95% CI = 4.8; 9.9, p-value < 0.001). Based on the logistic regression analysis, older males had a higher risk for the disease.

Conclusion: Our study finding might serve as a reference for clinicians when evaluating patients with cancer to search for any underlying condition that was not diagnosed during the disease course, as this will help to reduce the mortality associated with these conditions.

Citation Format: Muhammed Khaled Elfaituri, Sara Morsy, Nguyen Tien Huy. Incidence and mortality predictors of ill-defined illness (non-diagnosed conditions) as a cause of death in cancer patients: A population-based study [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 5763.
Abstract 5764: Descriptive epidemiology and clinicopathology of latent versus active JAK2-myeloproliferative neoplasms (MPN)
Authors: Haddad Philip A. , Gallagher Kevin , Hammoud Dalia
Published: September 2020

Splanchnic vein thromboses (SVT) encompass portal, mesenteric, splenic, and hepatic veins thromboses, and often cause Budd-Chiari syndrome (BCS). JAK2-MPNs were found to be the most common cause of SVT and BCS, accounting for up to 50% of all cases. Moreover, endothelial cells harboring JAK2 mutations have been demonstrated in patients with BCS which may contribute to the overall prothrombotic propensity. The aim of this study is to describe the epidemiology and clinicopathology of latent JAK2-MPN (LMPN), where JAK2 mutation is detected without clinical evidence of MPN, versus active JAK2-MPN (AMPN) where JAK2+ MPN is present, in a pooled cohort with SVT and/or BCS. In order to study the demographic characteristics, survival, and prognostic factors, we compiled a pooled database of 90 cases. Kaplan-Meier survival curves were constructed. Cox proportional-hazards model and Log-rank tests were used to assess the influence of demographic and clinical factors on survival. The cohort consisted of 65% LMPN and 35% AMPN. The median age for patients with SVT/BCS with LMPN and AMPN was 39(15-85) years and 42(24-80) years respectively. The peak incidence of SVT/BCS seems to occur between ages 36 and 48 for LMPN and 24-36 for AMPN. In both groups, females were twice more predominant than males. The AMPN group consisted of 41% Polycythemia Vera (PV), 22% Essential Thrombocythemia (ET), 28% Myelofibrosis (MF), and 3% others. There was no difference in the frequency of particular splanchnic veins' involvement or the number of coexisting acquired or inherited conditions between the two groups. Survival of patients with BCS did not differ by their MPN status or sex. Twenty-four percent of LMPN progressed to AMPN (57% ET, 21.5% PV, and 21.5% MF). The median time to progression was 55 (12-96) months among those that progressed. While the median time to progression was longer for males (63 vs 39 months) this difference did not reach statistical significance. This study describes for the first time the epidemiologic and clinical characteristics of patients with SVT/BSC and LMPN and compares them to those of AMPN. It also reveals that while some SVT/BCS with LMPN progress to active MPN diseases, a sizable proportion of these patients remain latent indicating that its prothrombotic propensity is independent of AMPN phenotype expression.

Citation Format: Philip A. Haddad, Kevin Gallagher, Dalia Hammoud. Descriptive epidemiology and clinicopathology of latent versus active JAK2-myeloproliferative neoplasms (MPN) [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 5764.
Abstract 5768: Peripartum blood transfusions are associated with increased risk of cancer: A national retrospective cohort study
Authors: Chae Young Kwang , Oh Michael S. , Lee Misuk , Cho Sukjoo , Park Jonghanne , Cho Geum Joon
Published: September 2020

Background: The effect of blood transfusions on the risk of developing primary cancer remains unclear, especially when administered in the peripartum period.

Methods: We conducted a retrospective cohort study of 270,529 pregnant women who delivered between January 1, 2007 and December 31, 2009, with data obtained from three national databases in South Korea. From this cohort, we identified 4,569 patients who received peripartum blood transfusions. We calculated hazard ratios (HRs) for new diagnoses of cancer, and adjusted them for relevant clinical factors using a Cox proportional hazards model.

Results: During follow-up, patients who received peripartum transfusions had an increased risk of developing cancer, with an adjusted HR of 1.16 (95% confidence interval [CI], 1.01-1.34). This risk was significant only among patients who received 3 or more units of blood, with an adjusted HR of 1.40 (95% CI, 1.10-1.79). Increased risk after transfusions were seen with brain, lung, ovarian, and gallbladder cancers. The difference in cancer risk between the transfusion and no-transfusion groups remained significant during both the first (1.34% vs 1.10%, p < 0.01) and second year (0.78% vs 0.58%, p < 0.01) after delivery.

Conclusion: Receipt of 3 or more blood transfusions in the peripartum period was associated with a significantly increased risk of developing cancer.

Citation Format: Young Kwang Chae, Michael S. Oh, Misuk Lee, Sukjoo Cho, Jonghanne Park, Geum Joon Cho. Peripartum blood transfusions are associated with increased risk of cancer: A national retrospective cohort study [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 5768.
Abstract 5790: An epigenome-wide gene-age interaction analysis reveals opposite effects of PRODH DNA methylation on survival between young and elderly early-stage NSCLC patients
Authors: Su Li
Published: September 2020

Introduction: Aging has become a global public health concern. The number of elderly lung cancer patients increased and elderly comprise the majority (81%) of non-small cell lung cancer (NSCLC) patients. DNA methylation is a potential prognostic biomarker for NSCLC, while it is also associated with aging. However, whether the association between DNA methylation and NSCLC survival is modified by aging is not known.

Methods: After quality control procedures, we used a two-stage study design to identify DNA methylation-age interaction that associated with overall survival (OS) for early-stage adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) patients from four international collaborating study centers. In the discovery phase, we investigated the interaction effect in LUAD and LUSC using Cox proportional hazards model, adjusted for covariates. We confirmed the significant interactions with Bonferroni adjusted P ≤ 0.05 were further confirmed in the validation phase using The Cancer Genome Atlas (TCGA) database.

Results: A total of 1,230 LUAD and LUSC patients were eligible for the study and 311,981 DNA methylation loci were evaluated. There were 613 (NLUAD = 492 and NLUSC = 121) patients in the discovery phase, and 617 (NLUAD = 332 and NLUSC = 285) patients in the validation phase, respectively. We identified one LUAD-specific CpG probe cg14326354PRODH, the effect on OS was significantly modified by age (HRinteraction = 0.989; 95% CI: 0.986-0.994; P = 9.18 × 10−7). The effect of DNA methylation on OS was different in young and elderly patients (young: HR = 2.44; 95% CI: 1.26-4.72; P = 8.34 × 10−3; elderly: HR = 0.58; 95% CI: 0.42-0.82; P = 1.67 × 10−3). Low methylation level benefited elderly LUAD patients. Moreover, there was an antagonistic interaction between low methylation of cg14326354PRODH and elder age (HRinteraction = 0.21; 95% CI: 0.11-0.40; P = 2.20 × 10−6).

Conclusion: Low methylation of cg14326354PRODH may benefit the survival of elderly LUAD patients, providing new insight into only age-specific prediction, but also potential methylation-specific drug targeting.

Citation Format: Li Su. An epigenome-wide gene-age interaction analysis reveals opposite effects of PRODH DNA methylation on survival between young and elderly early-stage NSCLC patients [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 5790.
Abstract 5796: Therapeutic effects of statins against lung adenocarcinoma via mutant p53 mediated apoptosis
Authors: Chou Cheng-wei , Lin Ching-Heng , Hsiao Tzu-Hung , Lo Chia-Chien , Hsieh Chih-Ying , Huang Cheng-Chung , Sher Yuh-Pyng
Published: September 2020

Introduction The p53 gene is well known as an important tumor suppressor gene. Mutant p53 genes account for about half of all lung cancer cases. There is increasing evidence for the anti-tumor effects of statins via inhibition of the mevalonate pathway. However, the cancer prevention and protection effects of statinsin population-based studies are controversial. Previous population-based studies have shown protective effects of statins in advanced lung cancer patients; however, the effects in early lung cancer patients are unclear.

Methods We investigated the correlation between statin use and lung cancer prognosis using the Taiwanese National Health Insurance Research Database retrospectively, mainly focusing on early-stage lung cancer. Cell viability, lipid raft changes, and motile activity from simvastatin-treated cell lines were examined to determine the effect of simvastatin under different p53 mutations. Apoptotic pathways and autophagy in lung cancer cells treated with simvastatin were analyzed using a western blot assay.

Results Statin treatments reduced the 5-year mortality (odds ratio, 0.43; 95% confidence interval [CI], 0.37-0.49;P< 0.001) in this population-based study. Simvastatin inhibited cell growth, especially in mutant p53 lung cancer cell lines. Significantly higher levels of cellular apoptosis and decreased lipid rafts were shown in mutant p53 lung cancer cells treated with simvastatin. Further, simvastatin increased activity of the caspase-dependent apoptotic pathway in lung cancer cells containing mutant p53. In addition, simvastatin reduced migration in cells with a mutated p53 gene.

Conclusion This study reports the protective effects of statin use in early-stage lung cancer patients regardless of chemotherapy. Simvastatin induced cellular apoptosis and regulated lipid raft content; further, motile activity was reduced in lung cancer cells with p53 missense mutations. These data suggest that statin use in selected lung cancer patients may have clinical benefits.

Citation Format: Cheng-wei Chou, Ching-Heng Lin, Tzu-Hung Hsiao, Chia-Chien Lo, Chih-Ying Hsieh, Cheng-Chung Huang, Yuh-Pyng Sher. Therapeutic effects of statins against lung adenocarcinoma via mutant p53 mediated apoptosis [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 5796.
Abstract 5837: Exploring integrated stress response pathway to target MYC-amplified medulloblastoma
Authors: Langman Sofya , Sorensen Poul H.
Published: September 2020

Medulloblastoma (MB) accounts for 20% of diagnosed brain tumors in children. The Group 3 (G3) MB subtype is found to be especially malignant. Current G3 MB treatment regimens provide for 50% 5-year survival rates and are associated with severe side effects, including hearing loss and neurocognitive deficits. G3 MB is characterized by MYC overexpression, which drives elevated protein translation in tumor cells. High expression levels of PERK correlate with poor survival outcome in G3 MB, and knockdown of PERK combined with hypoxic stress induces apoptosis in MB cells. PERK is an eIF2α kinase that responds to ER stress and is one of the key mediators of unfolded protein response. When ER stress is detected, PERK phosphorylates eIF2α to shut down global translation and to initiate integrated stress response. However, there are 3 other eIF2α kinases that work alongside PERK to help cells respond to various stresses. ISRIB is in an integrated stress response inhibitor that blocks P-eIF2α attenuation by eIF2B, thus maintaining high translation levels under stress. Combining ISRIB with stress such as hypoxia induces apoptosis in MB cells. In addition, combination of ISRIB and hypoxia is correlated with increased levels of oxidative stress. Apart from restoring global translation levels, ISRIB can prevent stress granules (SGs) from being formed. High expression of SG proteins such as G3BP1 and YB1 also correlates with poor survival outcome in G3 MB patients. When SG formation is inhibited by knocking down expression of G3BP1 and G3BP2, MB cells undergo higher levels of apoptosis under vincristine induced stress. We show that preventing G3 MB cells from undergoing integrated stress response and inhibiting formation of stress granules induces higher levels of cancer cell death. Overall, combining ISRIB with conventional chemotherapy may enhance the effects of the latter, which is increasingly relevant for patients suffering from G3 MB.

Citation Format: Sofya Langman, Poul H. Sorensen. Exploring integrated stress response pathway to target MYC-amplified medulloblastoma [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 5837.
Abstract 5849: Association between annexin-A6 expression and metabolic vulnerabilities of TNBC cells and their response to EGFR-tyrosine kinase inhibitors
Authors: Williams Stephen D. , Sakwe Amos M.
Published: September 2020

PURPOSE: Triple-negative breast cancer (TNBC), and the basal-like TNBC subtype, accounts for ~20% of all breast cancer (BC) cases, and is particularly prevalent in younger patients of African-American and Hispanic descent. The importance of annexin A6 (AnxA6) as a tumor suppressor in various cancers has been widely investigated, but the mechanisms by which AnxA6-induced basal-like triple negative breast cancer progression remain largely unknown. Physiological changes such as hypoxia, metabolic acidosis, and oxidative stress in the tumor microenvironment (TME) during cancer progression endow cancer cells with malignant properties, ultimately leading to metastatic dissemination that remains the major cause of death in cancer patients. By linking the altered expression status of AnxA6, with changes in metabolic output of the tumor cells and the efficacy of the otherwise ineffective epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), these metabolic vulnerabilities can be targeted to attenuate the growth and/or motility of this hard-to-treat breast cancer subtype. DESIGN METHODS: AnxA6-high (BT-549) and AnxA6-low (MDA-468) TNBC cell lines were cultured in hypoxic conditions (94%N2; 1%O2; 5%CO2; 37°C) using a Bactrox Hypoxia Chamber (Terra Universal Inc., Fullerton, CA). TNBC cells were treated with EGFR-TKI Lapatinib (Biovision Inc., Milpitas, CA).

RESULTS: We demonstrate that exposure of AnxA6-high (BT-549) and AnxA6-low (MDA-468) TNBC cell lines to acute hypoxia (1% O2; ≤24 hrs.) is associated with down-regulation of AnxA6 while chronic hypoxia (1% O2; >24 hrs.) led to a consistent up-regulation of AnxA6. AnxA6, FABP4, and HIF-1α downstream genes GLUT1 and PGK-1, mRNA levels were upregulated in chronic hypoxic conditions. AnxA6 metabolic studies reveal that contrary to AnxA6-high, down regulation of AnxA6 in TNBC cells increased lactate production, glucose consumption, and ATP-synthesis and lipid metabolism under hypoxia. In addition, chronic hypoxia sensitized TNBC cells to EGFR-TKIs. CONCLUSION: These data suggest that the transition from AnxA6-high to AnxA6-low may be dependent on hypoxia in the TME and consequently, TNBC cell survival and resistance to EGFR-targeted therapies.

Supported by NIH/NIGMS 1SC1CA211030 (AMS)

Citation Format: Stephen D. Williams, Amos M. Sakwe. Association between annexin-A6 expression and metabolic vulnerabilities of TNBC cells and their response to EGFR-tyrosine kinase inhibitors [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 5849.
Abstract 5910: Single-cell multiomic analysis of SNV, CNV, and protein expression
Authors: Ooi Aik , Mendez Pedro , Dhingra Dalia , Beard Nigel , Ruff David
Published: September 2020

Recent advancements in precision medicine, while highly promising, presents a major technical challenge to researchers due to disease heterogeneity. The emergence of single-cell technologies has greatly refined the resolution in which sample diversity can be investigated, enhancing the efficiency of selecting appropriate molecular targets. Additionally, applying multiomic analysis on single cells would further improve the understanding of cell-to-cell heterogeneity by providing unique insights on cellular and genetic composition. Using a two-step droplet microfluidic technology, the Mission Bio Tapestri Platform enables multiplex-PCR based high-throughput targeted DNA sequencing in single cells to obtain single-nucleotide variation (SNV) and copy number variation (CNV) information. By leveraging this technology, a new workflow is developed to detect protein expression in addition to DNA genotype in the same single cells. In this approach, cells are labeled with a pool of oligonucleotide-conjugated antibodies prior to loading the cells into the Tapestri Instrument for targeted DNA analysis. Sequencing libraries are then prepared from both antibody oligonucleotides and the amplified DNA sequences, followed by identification of single-cell DNA genotypes and protein signatures from the sequencing readout. In a mixed population of four cell lines, single-cell SNV and CNV information from 127 targeted amplicons and the protein data from 10 antibodies independently classified the cells into appropriate clusters. This method has been successfully performed on clinical samples with myeloid malignancies. In an acute myeloid leukemia (AML) sample, combined single-cell SNV, CNV, and protein expression data illustrated the heterogeneity within the sample. The data clearly identified CD3+ T cells and CD19+ B cells without pathogenic SNVs and CNVs. CD34hiCD11blo and CD34loCD11bhi subpopulations were also identified within the cells carrying the same pathogenic SNVs and CNVs. We believe that this novel multiomic technology will enable new discoveries in the complex relationship between genotype and phenotype, leading to a better understanding of disease biology, and subsequently better design of diagnostics and therapies.

Citation Format: Aik Ooi, Pedro Mendez, Dalia Dhingra, Nigel Beard, David Ruff. Single-cell multiomic analysis of SNV, CNV, and protein expression [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 5910.
Abstract 5956: New molecular mechanisms of small cell lung cancer growth
Authors: Alam Sk. Kayum , Wang Li , Ren Yanan , Hernandez Christina E. , Kosari Farhad , Roden Anja C. , Yang Rendong , Hoeppner Luke H.
Published: September 2020

Lung cancer is the leading cause of cancer death among both men and women in the United States. Small cell lung cancer (SCLC) results in over 30,000 deaths of Americans annually and comprises 15-17% incidences of lung cancer. SCLC is the most aggressive form of lung cancer due to its rapid doubling time and early widespread metastasis. Most SCLC patients present with advanced disease, respond to initial systemic chemotherapy, and then treatment refractory progression usually occurs within one year due to acquired drug resistance. Consequently, the median survival time of SCLC patients is only 9 to 20 months and merely 7% of SCLC patients survive beyond five years. A better molecular understanding of SCLC is necessary to develop much needed new treatment strategies. For the first time, we demonstrate that dopamine and cAMP-regulated phosphoprotein, Mr 32000 (DARPP-32) and its splice variant t-DARPP stimulate tumors of neuroendocrine origin, and we show DARPP-32 proteins are aberrantly overexpressed in human SCLC. Upregulation of t-DARPP protein in a subset of SCLC patients positively correlates with overexpression of genes associated with Notch signaling, including achaete-scute homologue 1 (ASCL1). We reveal that DARPP-32 isoforms are transcriptionally activated by ASCL1 in human SCLC cells. To determine whether DARPP-32 drives SCLC growth in vivo, we utilized an orthotopic lung cancer xenograft mouse model. Mice challenged with DARPP-32-ablated DMS-53 cells show a substantial decrease in lung tumor growth relative to controls. Correspondingly, we demonstrate increased tumor growth in mice harboring lung tumors formed by administration of DMS-53 or H1048 cells overexpressing DARPP-32 and t-DARPP proteins. Mechanistically, we describe how DARPP-32 isoforms promote SCLC growth through increased proliferation, Akt/Erk-mediated survival, and anti-apoptotic signaling. Taken together, we define new regulatory mechanisms of SCLC oncogenesis that suggest DARPP-32 isoforms may represent a negative prognostic indicator for SCLC and serve as a potential target for the development of new therapies.

Citation Format: Sk. Kayum Alam, Li Wang, Yanan Ren, Christina E. Hernandez, Farhad Kosari, Anja C. Roden, Rendong Yang, Luke H. Hoeppner. New molecular mechanisms of small cell lung cancer growth [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 5956.
Abstract 606: Cisplatin disrupts MDM2-DAXX-HAUSP complex, & induces apoptosis in acute leukemia cells
Authors: Tchounwou Paul B. , Kumar Sanjay
Published: September 2020

Cisplatin (CDDP) is an antitumor drug successfully used in the treatment of various types of human malignancies. It enhances toxicity of arsenic trioxide (ATO) in head and neck squamous cell carcinoma cell, small cell lung cancer, ovarian cancer cells, and oral squamous cell carcinoma cells (, and chronic myelogenous leukemia cells. CDDP induces cytotoxicity, DNA damage, oxidative stress, and apoptosis in acute promyelocytes leukemia (APL) cells. However, the molecular mechanisms of CDDP-induced P53 activation, cell cycle arrest, and apoptosis in APL cells remain poorly understood. We hypothesized that CDDP inhibits proliferation of APL cells through MDM2-DAXX-HAUSP complex disruption, p53 activation leading to cell cycle arrest and apoptosis. To test hypothesis, we used three APL cell lines and a mice model of APL and investigated CDDP effects on cells growth, complex disruption, cell cycle progression, and apoptosis by applying western blotting, immunoprecipitation(IP), gene knockdown techniques, immune/histocytochemistry, and confocal imaging. We found that CDDP induced stress signal transmitted by protein kinase (ATM, ATR) and its downstream targets (CHK1 & CHK2) phosphorylation leading to down-regulation of complex molecules expression and accumulation of p53. CDDP-induced p53 expression modulated cell cycle progression and apoptosis in APL cells, and stimulated the formation of more promyelocytes with dense granules through reduction of MDM2 expression in bone marrow cells. CDDP also activated p53 in APL mice liver tissues. This novel mechanism of action of CDDP in APL cells may help in the design of new anti-leukemic drugs for treatment of APL patients.

Citation Format: Paul B. Tchounwou, Sanjay Kumar. Cisplatin disrupts MDM2-DAXX-HAUSP complex, & induces apoptosis in acute leukemia 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 606.
Abstract 6073: The lipogenic regulator SREBP induces Transferrin in circulating melanoma cells, suppressing their susceptibility to ferroptosis
Authors: Hong Xin , Roh Whijae , Sullivan Ryan J. , Wong Keith H. , Wittner Ben S. , Guo HongShan , Dubash Taronish D. , Sade-Feldman Moshe , Wesley Ben K. , Boland Genevieve M. , Marvin Dieuwke L. , Bonesteel Todd , Lu Chenyue , Horwitz Elad , Aguet François , Freeman Samuel S. , Calhoun Katherine , Jewett Michelle K. , Nieman Linda T. , Hacohen Nir , Näär Anders M. , Ting David T. , Toner Mehmet , Stott Shannon L. , Getz Gad , Maheswaran Shyamala , Haber Daniel A.
Published: September 2020

Circulating tumor cells (CTCs) are shed by cancers into the bloodstream, where a viable subset overcomes oxidative stress to initiate metastatic outgrowth. Clonally derived cultured CTCs from patients with BRAF-mutant melanoma reveal upregulation of lipogenesis and iron homeostasis pathways, correlated with their baseline and acquired drug resistance. In CTCs, the lipogenesis regulator SREBP directly induces transcription of the iron carrier Transferrin (TF), thereby reducing intracellular reactive oxygen species (ROS) and lipid peroxidation, and conferring resistance to BRAF inhibitors and inducers of ferroptosis. Knockdown of endogenous TF impairs tumorigenesis by melanoma CTCs; their associated soft agar clonogenic defect is rescued by the lipophilic anti-oxidants Ferrostatin-1 or Vitamin E, and by cholesterol. Single cell RNA-seq of patient-derived melanoma CTCs identifies a subset with high lipogenic, iron metabolic and proliferative signatures, correlated with adverse clinical outcome. Together, the coordinated regulation of these SREBP-driven pathways contributes to cancer progression, drug resistance and metastasis.

Citation Format: Xin Hong, Whijae Roh, Ryan J. Sullivan, Keith H. Wong, Ben S. Wittner, HongShan Guo, Taronish D. Dubash, Moshe Sade-Feldman, Ben K. Wesley, Genevieve M. Boland, Dieuwke L. Marvin, Todd Bonesteel, Chenyue Lu, Elad Horwitz, François Aguet, Samuel S. Freeman, Katherine Calhoun, Michelle K. Jewett, Linda T. Nieman, Nir Hacohen, Anders M. Näär, David T. Ting, Mehmet Toner, Shannon L. Stott, Gad Getz, Shyamala Maheswaran, Daniel A. Haber. The lipogenic regulator SREBP induces Transferrin in circulating melanoma cells, suppressing their susceptibility to ferroptosis [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 6073.
Abstract 6098: Gut microbiome predicts response to immunotherapy in advanced NSCLC
Authors: Yang Lu , Yin Huihui , Wang Yan , Jiao Yuchen
Published: September 2020

Introduction: It has demonstrated that gut microbiome could affect the response to anti-programmed death 1 (PD-1) immunotherapy in the Western population. And one study has suggested that the diversity of gut microbiome was associated with responses to immunotherapy in China. However, there were some conflicts in these previous studies. This study is aimed to further explore the relationship between gut microbiome and clinical response in advanced non-small cell lung cancer (NSCLC) patients.

Methods: From 30th/January/2018, advanced NSCLC patients with anti-PD-1 immunotherapy were enrolled. Fecal samples were collected prior to starting therapy and serially after immunotherapy. Total DNA was extracted and 16S ribosome RNA gene sequencing was applied to assess gut microbiome composition.

Results: Clinical response rate was determined by radiography according to Response Evaluation Criteria in Solid Tumor Version 1.1. There were sixteen responders (referred to as R, thirteen with stable disease and three with partial response), and thirteen non-responders (NR), yielding a response rate of 55.2%. Firmicutes, Bacteroidetes, Actinobacteria and Proteobacoteria were the main microbial composition in baseline stool samples. Gut microbiome diversity at baseline was not statistically different between responders and non-responders. However, the relative abundance of Akkermansia genus at baseline was significantly enriched in R cohort (p=0.0363). The count of OTUs at progression increased significantly during the treatment. (p=0.012). Of note, Alloprevotellal and Capnocytophaga were overrepresented significantly at progression, indicating that they were associated with unfavorable clinical outcome.

Conclusions: This study illustrated that the relationship between clinical outcomes and gut microbiome by dynamic monitoring. Akkermansia present at baseline feces was related with better clinical response to immunotherapy. The increase of some bacteria abundance, especially Alloprevotellal and Capnocytophaga, may predict disease progression during the immunotherapy. This study will continue to collect fecal samples and further verify these conclusions.

Citation Format: Lu Yang, Huihui Yin, Yan Wang, Yuchen Jiao. Gut microbiome predicts response to immunotherapy in advanced NSCLC [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 6098.
Abstract 6152: Characterizing cancer stem cells in diffuse intrinsic pontine glioma
Authors: Ferris Sarah F. , Surowiec Rachel K. , Espinoza Carlos , Galban Stefanie
Published: September 2020

Diffuse Intrinsic Pontine Glioma (DIPG) is a lethal pediatric brain cancer with a five-year survival rate of less than 1%. This is reflective of a 100% tumor reoccurrence emphasizing the need for efficacious therapies. Previous studies have identified Aldehyde dehydrogenase expression (ALDH) as a marker for stemness and an indicator of tumor aggression and therapeutic resistance in many malignancies including adult glioma. Understanding the role of cancer stem-cells in gliomagenesis of DIPG is critical for developing therapies targeting resistance and averting 200-400 pediatric deaths in the US per year. We identified a subpopulation of cells expressing the ALDH cancer stem-cell marker in patient-derived DIPG cell lines with high variability in ALDH expression among the cell lines likely indicating previous treatment and resistance. As expected, proliferation and neurosphere formation as well as neurosphere size were dramatically increased in ALDH+ CSC compared to ALDH- CSC supporting our hypothesis of a tumor initiating, aggressive subpopulation likely conferring therapeutic resistance. To evaluate tumor initiation properties of ALDH+ CSC subpopulation in vivo, implantation of SU-DIPG-7 luciferase expressing cells into the pons of immunocompromised mice (NSG) was performed after sorting for ALDH by FACS. Our results demonstrate tumor initiation and tumor growth in ALDH+ mice as assessed by Bioluminescence imaging (BLI) as well as histological analysis. Furthermore, survival of mice implanted with ALDH+ CSCs was significantly reduced compared to mice implanted with ALDH- cells indicating tumor initiating and ‘stem cell' properties of this subpopulation. In summary, we have identified and characterized ALDH+ cancer stem-cells in DIPG which are more aggressive than the non-cancer stem-cell population (ALDH-). These results highlight the need to develop therapies targeting this highly aggressive cancer stem-cell population. Future studies investigating the gene regulation response to treatment in these stem-cell subpopulations could provide valuable insight to co-targeted therapies to address treatment resistance in DIPG.

Citation Format: Sarah F. Ferris, Rachel K. Surowiec, Carlos Espinoza, Stefanie Galban. Characterizing cancer stem cells in diffuse intrinsic pontine glioma [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 6152.
Abstract 620: The response of human glioblastoma cells to the novel drug 1A-116 is dependent on the circadian clock
Authors: Trebucq Laura Lucia , Cardama Goergina A. , Menna Pablo Lorenzano , Golombek Diego A. , Chiesa Juan J. , Marpegan Luciano
Published: September 2020

Glioblastomas (GBM) account for almost half of the malignant central nervous system tumors and have the highest mortality rates with only about 5% of the adult patients surviving 5 years after diagnosis. Current therapy for GBM is generally very aggressive, combining chemotherapy, radiotherapy, and surgical removal and increased the life expectancy only from 11 to 14 months, but, almost no significant advances were made in the last decades. Taking this into account, more effective compounds and new strategies, such as chronotherapeutic ones, need to be developed.

The novel drug 1A-116 specifically blocks the interaction of the Rho GTPase Rac1 with some of its activators (GEFs proteins, such as Tiam1) inhibiting cell motility and proliferation of GBM. To optimize the effectiveness of 1A-116 a chronotherapeutic approach was implemented in vitro by applying the drug at different circadian times to synchronized LN-229 human GBM cells. Cell cultures displayed a functional circadian clock measured by Bmal1-luc activity and BMAL1 protein expression. Expression of Tiam1 was also assessed, finding circadian expression rhythms. The effects of 1A-116 on either cell proliferation, motility and toxicity, were dependent on the circadian cellular clock. The preliminary in vivo studies consisted on treating nude mice with 1A or control at ZT3 or 12. We found that the median survival of the mice treated at ZT3 was 82 days and at ZT12 was 78 days. These results suggest that a chronopharmacological approach based on the drug 1A-116 is a feasible strategy to improve GBM treatment outcomes.

Citation Format: Laura Lucia Trebucq, Goergina A. Cardama, Pablo Lorenzano Menna, Diego A. Golombek, Juan J. Chiesa, Luciano Marpegan. The response of human glioblastoma cells to the novel drug 1A-116 is dependent on the circadian clock [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 620.
Abstract 6239: A novel topical delivery system of carvedilol for skin cancer prevention
Authors: Chen Mengbing , Yeung Steven , Shamim Md Abdullah , Coronel Kaitlyn , Shahid Ayaz , Andresen Bradley , Wang Jeffrey , Huang Ying
Published: September 2020

Solar ultraviolet (UV) light-induced non-melanoma skin cancer is the most common type of cancer diagnosed in the United States and remains one of the major public health concerns. There is an urgent need to develop effective and safe skin cancer chemopreventive agents. We have previously demonstrated that carvedilol, an FDA approved β-blocker for cardiovascular diseases, could prevent UV-induced skin cancer in vitro and in vivo. However, the β-adrenergic blockade effects of carvedilol might be a barrier for its repurposing as a cancer chemopreventive agent. In this study, we designed a novel topical formulation, namely carvedilol-loaded transfersomes (T-CAR), to maintain cancer preventive properties of carvedilol while avoiding systemic cardiovascular effects. T-CAR formulations were prepared using different phospholipids and surfactants at various ratios by the thin-film hydration method. The formulations were characterized in terms of particle size, zeta potential, and encapsulation efficiency. An optimal formulation was identified that composes carvedilol, soy phosphatidylcholine (SPC), and Tween 80 at a ratio of 1:3:0.5 with the particle size of 95.84 ± 3.8 nm, the zeta potential of +11.34 ± 0.7 mV, and the encapsulation efficiency of 93.7 ± 5.1%. The in vitro drug release and ex vivo skin permeation studies were performed using Pur-A-Lyzer mini dialysis kit and Franz diffusion cells, respectively. The optimal formulation showed a relative slow in vitro release profile and it could permeate through the porcine ear skin to the same degree as free carvedilol dissolved in acetone. 200 uL of 10 uM carvedilol dissolved in acetone, T-CAR containing the same amount of carvedilol as in acetone group, or the plain transfersome were topically applied to SKH-1 hairless mice 30 minutes before and immediately after single-dose UV irradiation. Compared to carvedilol solution, T-CAR demonstrated the same inhibitory effects against UV-induced epidermis thickening but exhibited increased inhibitory effects against UV-induced cyclobutane pyrimidine dimer (CPD) formation and mRNA expression of pro-inflammatory cytokines IL-1β and IL-6. In addition, carvedilol was not detectable by LC-MS in plasma after topical administration. These findings suggest that T-CAR could be a promising topical carvedilol formulation to prevent UV-associated skin cancer, with enhanced photoprotective effects but neglectable systemic cardiovascular effects.

Citation Format: Mengbing Chen, Steven Yeung, Md Abdullah Shamim, Kaitlyn Coronel, Ayaz Shahid, Bradley Andresen, Jeffrey Wang, Ying Huang. A novel topical delivery system of carvedilol for skin cancer prevention [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 6239.
Abstract 6244: Turning poorly vascularized tumors into highly vascularized tumors with nanoparticles: Proof of concept and pharmacometric analysis
Authors: Sicard Guillaume , Rodallec Anne , Correard Florian , Vaghi Cristina , Poignard Clair , Ciccolini Joseph , Benzekry Sébastien , Sergé Arnauld , Fanciullino Raphaelle
Published: September 2020

Combinatorial regimen are a mainstay in oncology. Taxanes and trastuzumab are both gold standards for HER2 positive breast cancer. To what extent they could reshape tumor micro-environment remains to be elucidated. The aim of this study was to compare in breast cancer mice models the antiproliferative efficacy and impact on tumor neo-angiogenesis of trastuzumab stealth immunoliposomes encapsulating docetaxel and stealth liposomal docetaxel associated with free trastuzumab. Nude mice bearing human MDA-MB-231 breast cancer tumors were treated by saline (control), docetaxel liposome (5mg/kg) associated with free trastuzumab (1.9 mg/kg) or trastuzumab immunoliposome (eq. 5 mg/kg docetaxel and 1.9 mg/kg trastuzumab) QW for 4 consecutive weeks. Tumor growth was monitored twice a week by volumetric measurement using a caliper. Blodd vessels density was evaluated in vivo by fluorescence imaging using Angiosense as a vascular probe plus ex-vivo using anti-CD31 antibodies and electronic microscopy at 2, 4 and 6 weeks. A specific algorithm was developed on MatLab for imaging analysis. No treatment-induced toxicities was observed. After 4 weeks of dosing, no difference in efficacy was seen between treatments (p=0.798) and Angiosense imaging did not show differences either (p=0.628). However electronic microscopy showed that both immunoliposomes and liposomes increased tumor vascular density as compared with control. In addition, a difference between treatments was evidenced because a marked increase in vascularization of central tumors VS. peripheral tumors was observed with trastuzumab immunoliposome as compared with standard docetaxel liposome (p=0.00023). A numerical model based on porous medium theory combined with multi-scale approaches has been derived to describe the spatial distribution of the drugs within tissues. Overall our data suggest that trastuzumab stealth immunoliposomes induce a normalization of tumor vessels with an increase in blood vessels density in central tumors. This could pave the way for sequencing combinatorial strategies so as to improve drug delivery into tumors of the associated treatments.

Citation Format: Guillaume Sicard, Anne Rodallec, Florian Correard, Cristina Vaghi, Clair Poignard, Joseph Ciccolini, Sébastien Benzekry, Arnauld Sergé, Raphaelle Fanciullino. Turning poorly vascularized tumors into highly vascularized tumors with nanoparticles: Proof of concept and pharmacometric 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 6244.