bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 980: Maligant ascites microenvironment skews M2/M1 macrophage polarization in ovarian cancer during peritoneal metastasis
Authors: Wang Huogang , Yung Mingo MH , Chen Arvin FS , Ngan Hextan YS , Chan David W.
Published: September 2020

Advanced ovarian cancers with peritoneal metastases are usually accompanied with metastatic recurrence and high mortality. Macrophages, as a primary cell population in the tumor microenvironment, can be active and polarized into heterogeneous tumor-associated macrophages (TAMs). Clinical evidence indicates that a high M2/M1 ratio of TAMs are commonly found in malignant ascites microenvironment (MAM), and is attributed to a poor prognosis in advanced ovarian cancers. However, the molecular mechanisms of macrophage polarization in MAM remain elusive. Here, we report that MAM could modulate Hippo and/or Wnt signaling cascades in facilitating M2 macrophage polarization. In this study, we firstly evidenced the heterogeneity macrophage populations in cancerous omental tissues and ascites cancer cell spheroids of ovarian cancers by multiplex immunohistochemical, flow cytometry and immunofluorescent analyses. Consistently, in vitro polarization model demonstrated that co-cultured monocyte (THP-1 and U937) with omentum conditioned medium (OCM) or ascites significantly led to more CD206+ M2 macrophage than CD86+ M1 macrophage or M2/M1 ratio. Quantitative RT-PCR (qPCR) analysis showed that TAM-related genes: CCL2, IL6, and IL10 genes were significantly upregulated in OCM treated macrophage. Intriguingly, OCM treated macrophage could significantly promotes ovarian cancer cell migratory capacities. Importantly, RNA-seq analysis identified Hippo and Wnt signaling cascades may play important roles in macrophage polarization in malignant OCM and ascites. Further investigations of these pathways in M2/M1 macrophage polarization in MAM are warranted.

Citation Format: Huogang Wang, Mingo MH Yung, Arvin FS Chen, Hextan YS Ngan, David W. Chan. Maligant ascites microenvironment skews M2/M1 macrophage polarization in ovarian cancer during peritoneal metastasis [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 980.
Abstract 99: Phenotypic characterization of knockdown of CDKs in the intestinal epithelial cells
Authors: Lu Shuyan , Hu Wenyue , Sung Tae , Hirakawa Brad , Amaro Marina , Jessen Bart
Published: September 2020

Cyclin-dependent kinases (CDKs) are serine/threonine kinases that bind to cyclins and together act as regulators of cell cycle progression. CDK inhibitors have various therapeutic potentials including cancer (e.g. CDK4/6 inhibitors). Intestinal epithelial cells are highly proliferative; however, the impact of individual CDK inhibition on intestinal cell proliferation has not been well studied. The IEC6 cell line, a non-transformed rat small intestinal epithelial cell line with characteristics of crypt epithelial cells, was utilized in the current study to understand the role of CDKs in the proliferation and survival of intestinal cells. In the initial experiment CDK4/6 inhibitor palbociclib treatment demonstrated a lack of potent toxicity in the IEC6 in comparison with MCF7 (breast cancer cell line), indicating the absence of intestinal cell reliance on CDK4 or CDK6 for cell cycle progression. To further illustrate the role of CDKs in intestinal cells, targeted gene knockdown experiments using CDK1, 2, 4, and 6 siRNA and lipofectamine-mediated transfection were conducted. Surprisingly, only CDK1 knockdown causes profound cell death. CDK2, 4, or 6 knockdowns, whether single, double or triple combinations, did not have significant impact on IEC6 cells, as measured by multiple endpoints including impedance, ATP viability, and caspase activity. To further understand the role of CDK1 in cell cycle, various seeding densities of IEC6 cells were used with CDK1 siRNA. Although the impact of CDK1 knockdown on IEC6 cells decreased at the initial proliferation phase with the higher seeding density, there was still a decrease of viability observed at the later time point (when cells reached proliferation plateau), indicating a non-canonical role of CDK1 that is not related to cell cycle. This research supports the concept that CDK1, but not CDKs 2, 4, or 6, is essential for intestinal cell cycle progression and explains the lack of GI toxicity observed with palbociclib.

Citation Format: Shuyan Lu, Wenyue Hu, Tae Sung, Brad Hirakawa, Marina Amaro, Bart Jessen. Phenotypic characterization of knockdown of CDKs in the intestinal epithelial 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 99.
Abstract 992: IL-6/STAT3-mediated inflammatory signaling results in dysregulated fibroblast growth factor receptor activation and tumor cell signaling in human pancreatic adenocarcinoma
Authors: Nawalaniec Karol , Wang Yuxiang , Moench Romana , Cao Minghua , Luo Yueming , Hsiao Li-Li , Waaga-Gasser Ana Maria
Published: September 2020

Pancreatic ductal adenocarcinoma (PDAC) is a devastating malignancy characterized by delayed diagnosis and the lowest five-year survival amongst all cancer sites. PDAC development and progression are attributed to chronic inflammation and associated tissue repair mechanisms which contribute to high density tumor formation and chemoresistance. PDAC-associated inflammation is, in part, owed to high levels of interleukin 6 (IL-6) and its activated pathways including JAK/STAT3. In Addition, Fibroblast growth factor (FGF) signaling pathways are known to be dysregulated in cancer and have been implicated in various malignancies including PDAC. As such, we sought to explore the association between IL-6-induced inflammatory signaling and FGF signaling activation. To observe this effect as it pertains to carcinogenesis and cancer progression we used four human cell lines ranging from healthy pancreatic ductal epithelial (HPDE) cells to various grade (G) PDAC cells. Cells were treated with IL-6 at a concentration of 100 ng/mL for 24 hours. The results showed increased phosphorylation of STAT3 in BxPC-3 (G2) and PANC-1 (G3) cells with no change noted in HPDE and SW1990 (G2, metastatic) cells. IL-6 stimulation likewise resulted in markedly increased phosphorylation of the fibroblast growth factor receptor substrate 2-alpha (FRS2-α) and its downstream component Erk1/2 in BxPC3 cells, indicating activation of FGF signaling through the MAPK/Erk pathway in our low grade PDAC cell model. In conclusion, the results demonstrate a potential link between IL-6 induced inflammatory signaling and dysregulation of FGF signaling in human PDAC. Activated STAT3 signaling seems to be involved in this process. Existing cancer therapies aimed at controlling growth factor signaling broadly target vascular endothelial or epithelial growth factor receptors (VEGF/EGFR) with marginal efficacy and adverse side effects in PDAC. Identifying the pivotal link between inflammatory signaling and dysregulation of fibroblast growth factor signaling will allow for the development of more precisely targeted and effective cancer therapy in pancreatic cancer. This may minimize clinically observed exaggerated fibrotic events within the tumor and may hence result in higher vulnerability for established chemotoxic therapies in the tumor cells.

Citation Format: Karol Nawalaniec, Yuxiang Wang, Romana Moench, Minghua Cao, Yueming Luo, Li-Li Hsiao, Ana Maria Waaga-Gasser. IL-6/STAT3-mediated inflammatory signaling results in dysregulated fibroblast growth factor receptor activation and tumor cell signaling in human pancreatic adenocarcinoma [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 992.
Abstract CT074: Pre-existing ESR1 mutations in early-stage primary breast cancer predict failure of endocrine therapy and poor survival
Authors: Dahlgren Malin , George Anthony M. , Brueffer Christian , Gladchuk Sergii , Chen Yilun , Vallon-Christersson Johan , Hegardt Cecilia , Häkkinen Jari , Rydén Lisa , Malmberg Martin , Larsson Christer , Gruvberger-Saal Sofia K. , Ehinger Anna , Loman Niklas , Borg Åke , Saal Lao H.
Published: September 2020

Background: More than three-quarters of primary breast cancers are positive for estrogen receptor alpha (ER; encoded by the gene ESR1), the most important factor for directing anti-estrogenic endocrine therapy. Although mutation in ESR1 is known as an acquired mechanism of resistance to endocrine therapy (ET), found in 12-55% of metastatic breast cancers treated previously with ET, the impact of ESR1 mutation on therapy response in primary breast cancer is unclear. Patients and methods: In this study we analyzed 3217 real-world and population-based early-stage primary breast cancers (within the SCAN-B study, ClinicalTrials.gov NCT02306096). Tissues were sampled from initial diagnosis prior to any treatment and analyzed for the presence of ESR1 mutations using RNA sequencing. Mutations were verified by SAGAsafe droplet digital PCR. Results: We identified ESR1 resistance mutations in 30 cases (0.9%), of which 29 were ER-positive (1.1%). In ER-positive disease, presence of ESR1 mutation was significantly associated to poor relapse-free survival (RFS) and overall survival (OS) (p=0.011 and p=0.019, respectively), and moreover predicted poor RFS and OS within the patient group that received ET (p=0.007 and p=0.010, respectively). Conclusions: These results indicate that ESR1 mutations at diagnosis of untreated primary breast cancer are rare, however we confirm for the first time that such early mutations predict eventual resistance to standard hormone therapy in the adjuvant setting. If replicated, tumor ESR1 screening may be considered in ER-positive primary breast cancer and, in mutated cases, ER-degraders such as fulvestrant or other therapeutic options may be considered as more appropriate.

Citation Format: Malin Dahlgren, Anthony M. George, Christian Brueffer, Sergii Gladchuk, Yilun Chen, Johan Vallon-Christersson, Cecilia Hegardt, Jari Häkkinen, Lisa Rydén, Martin Malmberg, Christer Larsson, Sofia K. Gruvberger-Saal, Anna Ehinger, Niklas Loman, Åke Borg, Lao H. Saal. Pre-existing ESR1 mutations in early-stage primary breast cancer predict failure of endocrine therapy and poor survival [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 CT074.
Abstract CT152: First in human study with EOS100850, a novel potent A2A antagonist, shows excellent tolerance and clinical benefit in immune resistant advanced cancers
Authors: Buisseret Laurence , Rottey Sylvie , de Bono Johann , Mossakowski Manon , Delafontaine Brant , Manickavasagar Thubeena , Kotecki Nuria , Martinoli Chiara , Schneider Manfred , De Henau Olivier , Chevron Patricia , Lager Joanne , Machiels Jean-Pascal
Published: September 2020

Adenosine is a strong immunosuppressive mediator often found at high extracellular levels in the microenvironment of most tumors. Blocking A2A receptors, predominantly expressed on tumor-infiltrating immune cells, is aimed at reversing the immunosuppressive effect of adenosine. EOS100850 is a potent and highly selective small molecule antagonist of the A2A receptor that remains active even at the high adenosine concentration found in tumors and that does not cross the blood-brain barrier. This multicenter FIH Phase I clinical trial (NCT02740985) assessed the safety, pharmacokinetic (PK), pharmacodynamic (PD), and preliminary clinical activity of EOS100850 monotherapy in subjects with refractory solid tumors. Pharmacodynamic assessments were performed in peripheral blood and tumor biopsies with simultaneous determination of plasma PK. Twenty-one adult participants with advanced malignancies were treated with EOS100850 monotherapy. Cohorts of 3 participants each were evaluated at 20 and 40 mg QD or 40 mg BID administration. Six participants were subsequently tested at doses of 80 mg BID and at the maximum administered dose of 160 mg BID. All subjects had at least 1 prior regimen (median 3). PK analysis indicated dose-proportional increase of EOS100850 plasma exposures up to 80 mg BID while PD evaluations revealed prolonged and maintained A2AR inhibition over the dosing interval. The most common adverse events (>15%) attributed to the investigational treatment included Grade 1-2 nausea, fatigue, vomiting, and liver enzyme abnormalities. No drug-related Grade 3-4 AEs and no dose-limiting toxicities were observed. As of the data cut off (15JAN20), stable disease was observed in 6 participants who remain on therapy, including head & neck cancers, endometrial cancers, and checkpoint inhibitor-refractory melanoma. Further evaluation is ongoing as monotherapy and in combination with pembrolizumab or chemotherapy in both immune checkpoint-naïve and -refractory patients.

Citation Format: Laurence Buisseret, Sylvie Rottey, Johann de Bono, Manon Mossakowski, Brant Delafontaine, Thubeena Manickavasagar, Nuria Kotecki, Chiara Martinoli, Manfred Schneider, Olivier De Henau, Patricia Chevron, Joanne Lager, Jean-Pascal Machiels. First in human study with EOS100850, a novel potent A2A antagonist, shows excellent tolerance and clinical benefit in immune resistant advanced cancers [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 CT152.
Abstract CT186: HEPANOVA: Interim safety analysis from a phase 2 study of Tumor Treating Fields (TTFields, 150 kHz) concomitant with sorafenib in advanced hepatocellular carcinoma (HCC)
Authors: Gkika Eleni , Brunner Thomas , Thimme Robert , Grosu Anca-Ligia
Published: September 2020

Background: Tumor Treating Fields (TTFields) are an anti-mitotic, regional treatment modality, utilizing low intensity alternating electric fields delivered non-invasively to the tumor using a portable medical device. TTFields are approved by the FDA for the treatment of glioblastoma and malignant pleural mesothelioma. In vitro and invivoTTFields at 150 kHz reduced the proliferative potential of hepatocelluar carcinoma (HCC) cell lines. Most HCC patients are not amenable to curative therapies and are treated with systemic chemotherapy following failure of local treatment options. The HEPANOVA Phase II trial (NCT03606590) is a prospective, single-arm study investigating the safety and efficacy of TTFields as a treatment option for advanced HCC concomitant with sorafenib in 25 patients. We present the interim saftey data of the HEPANOVA trial.

Methods: Incidence and severity of adverse events of the first nine patients who have been enrolled in the HEPANOVA Phase II trial were analyzed based on the MedDRA body system (system organ class) and preferred terms.

Results: The median age was 70 (range 63-85), all the patients were male, 3 (33.3%) patients had an ECOG score of 0. The median number of prior treatments was 2 (0-6) and median time from diagnosis was 9 (0-56) months. Five (55.6%) patients had Barcelona Clinic Liver Cancer (BCLC) stage B disease. The median treatment duration was 2 months for sorafenib and 3 months for TTFields. The total TTFields treatment duration for all nine patients was 29.1 months. Six (66.7%) patients had severe AEs (grade 3-4) and 3 (33.3%) had serious adverse events (SAE), none of which were related to TTFields. Mild-moderate skin toxicity was the only TTFields-related AE, reported in seven (78%) patients.

Conclusions: No unanticipated serious adverse device effects have been reported to date in the HEPANOVA study. The majority of AEs reported to date were expected and associated with the disease or chemotherapy. The most common adverse event reported was the expected TTFields-related dermatitis. In summary, these results emphasize the tolerably and safety of TTFields (150 kHz) concomitant with sorafenib in advanced hepatocellular carcinoma.

Citation Format: Eleni Gkika, Thomas Brunner, Robert Thimme, Anca-Ligia Grosu. HEPANOVA: Interim safety analysis from a phase 2 study of Tumor Treating Fields (TTFields, 150 kHz) concomitant with sorafenib in advanced hepatocellular carcinoma (HCC) [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 CT186.
Abstract CT187: Pembrolizumab and nanoparticle albumin bound paclitaxel (nab-paclitaxel) as salvage therapy for metastatic urothelial carcinoma (UC)
Authors: Giannatempo Patrizia , Raggi Daniele , Farè Elena , Marandino Laura , Bandini Marco , Colecchia Maurizio , Pederzoli Filippo , Calareso Giuseppina , Madison Russell , Monopoli Francesco , Russo Giusy , Ferrara Roberto , De Braud Filippo , Montorsi Francesco , Necchi Andrea
Published: September 2020

Methods: In an open-label, single-arm, phase 2 trial, pts receive pembro 200 mg intravenously (IV) on D1 and 125 mg/m2 IV nPtx on D1 and D8, every 3 weeks, until disease progression (PD) or onset of unacceptable toxicity. Key inclusion criteria are: predominant UC histology, failure of ≤2 platinum-based CT for metastatic disease. Response is evaluated by RECIST v.1.1 criteria every 2 cycles. Biomarkers include PD-L1 expression with the combined positive score (CPS) and comprehensive genomic profiling on tumor and blood samples (FoundationONE and FoundationACT assay). The primary endpoint of the study is the progression-free survival (PFS). The target is to detect an improvement in the median PFS from ≤3.0 months (H0) to ≥5.0 months (H1). Results: Between 01 and 12/2019, PEANUT study enrolled 63 pts evaluable for the study endpoint: 24% female, median age 69 y (IQR 61-73); 75% had failed 1 prior systemic therapy vs 25% 2 prior therapies; 35% had ECOG-performance status 1; 33% had liver metastases. The median TMB was 6.9 mut/Mb. After median follow-up of 5 months, 33 pts have relapsed (52%). The median PFS was 5 months (95%CI: 4-not achieved). The confirmed objective response-rate (ORR) was 44.4% (95%CI: 0.34 to 0.55): 19 partial responses and 9 complete responses (14.4%). Grade 3-4 treatment-related adverse events (TRAE) were seen in 14 pts (22%). Most common any-grade TRAE included alopecia (76%), neutropenia (33.3%), and asthenia (33%). Permanent interruption of treatment (nPtx only) was seen in 9 pts (14.2%) due to toxicity. Neither TMB nor CPS were significantly associated with PFS on univariable analyses.

Conclusions: Preliminary results from PEANUT study demonstrated a promising PFS and a clinically meaningful ORR in II-III line setting of advanced UC. Updated data, including mature OS, duration of response and biomarkers will be presented.

Citation Format: Patrizia Giannatempo, Daniele Raggi, Elena Farè, Laura Marandino, Marco Bandini, Maurizio Colecchia, Filippo Pederzoli, Giuseppina Calareso, Russell Madison, Francesco Monopoli, Giusy Russo, Roberto Ferrara, Filippo De Braud, Francesco Montorsi, Andrea Necchi. Pembrolizumab and nanoparticle albumin bound paclitaxel (nab-paclitaxel) as salvage therapy for metastatic urothelial carcinoma (UC) [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 CT187.
Abstract CT219: Efficacy of TTFields in elderly patients with newly diagnosed glioblastoma (GBM): Sub-group analysis of the EF-14 trial
Authors: Ram Zvi , Zhu Jay-Jiguang
Published: September 2020

Background: Tumor Treating Fields (TTFields) are an anti-mitotic, regional treatment modality, utilizing low intensity alternating electric fields delivered non-invasively to the tumor using a portable medical device. In the EF-14 phase 3 study leading to FDA approval, TTFields significantly extended survival in newly diagnosed GBM when added to maintenance temozolomide (TMZ). Elderly GBM patients usually have worse prognosis and often receive only partial treatment for the disease. The aim of the following post-hoc analysis was to examine the effects of TTFields in the elderly population (≥65 years of age) enrolled in the EF-14 study. Method: All 134 elderly patients (≥65 years of age) from the EF-14's intent-to-treat population were included in the analysis, Overall survival (OS) and progression-free survival (PFS), as well as adverse event frequency and severity were compared between the TMZ + TTFields arm and the TMZ alone arm. Result: The median age was 69 (range: 65-83), 69% were male. Median PFS from randomization was 6.5 months versus 3.9 months in the TTFields + TMZ Vs. TMZ alone arms, respectively (hazard ratio [HR], 0.47 [95%CI 0.30, 0.74] P<0.0236). Median OS was 17.4 months versus 13.7 months in the TTFields + TMZ versus TMZ alone arm, respectively (HR 0.51 [CI 0.33, 0.77] P<0.020). Serious adverse events (SAEs) were reported in 39% of patients treated with TTFields + TMZ and in 33% of patients treated with TMZ alone. None of the SAEs were considered related to TTFields. SAEs were considered related to TMZ or to the underlying disease. Grades 1-2 skin AEs were observed in 51% of patients. Conclusion: Consistent with the overall outcome of the EF-14 study, elderly patients treated with TMZ + TTFields showed significantly better OS compared to patients on TMZ alone, and without increase in grade 3-4 toxicity.

Citation Format: Zvi Ram, Jay-Jiguang Zhu. Efficacy of TTFields in elderly patients with newly diagnosed glioblastoma (GBM): Sub-group analysis of the EF-14 trial [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 CT219.
Abstract CT222: Colon cancer in elderly patients: Retrospective analysis of targeted therapy era
Authors: KIM JIN YOUNG , Shin Sojin , Heo Mi Hwa , Lee Kang Kook
Published: September 2020

Purpose: This study was conducted in order to evaluate the response rate, progression free survival (PFS), safety and prognostic factors of targeted combination chemotherapy in colon cancer patients older than 65 years. Methods: This retrospective study included 69 elderly patients (≥ 65 years) with metastatic Colon cancer who have not received palliative chemotherapy before. All patients were histologically confirmed colon cancer at Keimyung university Dongsan medical center from Jan 2007 to July 2017. All recruited patients received either cetuximab or bevacizumab every 2 weeks. Results: The median age was 73.35 (65-87) and 33 were right side, 32 were left, 2 were transverse and rest of 2 patients were multiple primary sites. Bevecizumab was administerd to 42 patients (60.9%) and cetuximab was administered to 27 patients (39.1%). Complete response was 9 patients (13%), partial response was 23 (33.3%) and progression disease was 9 (13.0%). Overall response rate was 31 (46.4%) and disease control rate was 42 (62.3%). Progression free survival was 12.038 ±1.565 months (95% CI: 8.97~12.12). The most common hematologic side effects was anemia with grade 3/4 in 14 (20.3%). Systemic chemotherapy with targeted agents were moderately tolerated with grade 3/4 neutropenia 40 (57.9%). Conclusion: In elderly patients with colon cancer, clinical outcomes showed similar results compared with prior clinical trials. Preventing such complications may improve survival.

Citation Format: JIN YOUNG KIM, Sojin Shin, Mi Hwa Heo, Kang Kook Lee. Colon cancer in elderly patients: Retrospective analysis of targeted therapy era [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 CT222.
Abstract LB-007: The emergence of tumor organoid engineering as a powerful tool for drug efficacy and mechanism of action profiling
Authors: Wang Fei , Zhou Rongyun , Dong Xinlong , Ma Yuqing , Zhou Jun , Xu Xiaoxi , Huang Yujun , Li Henry , Wang Phillip S-Z
Published: September 2020

Introduction: Patient-derived models such as organoids are a powerful translational tool for preclinical research with several advantages over current conventional models: 1) they are derived from stem cells with self-organizing and renewal properties reflecting the 3D structure and morphology of original tissue; 2) they preserve the original genomic, physiological, and multicellular identity thereby representing patient diversity and heterogeneity in a dish; 3) they provide in vitro convenience for greater throughput for screening large numbers of compounds and/or patients; 4) they are amenable to genetic engineering. Methods: We have used Hubrecht Organoid Technology (HUB) protocols to systematically create a biobank of organoids derived from a well-annotated patient-derived xenograft (PDX) library (the world's largest commercially available PDX library with over 2,500 models, covering a variety of carcinomas, with extensive pathology, genomic, and treatment information), referred to as PDX-derived organoids or PDXOs. Results: We have currently developed more than 200 PDXOs covering many cancer types, including colorectal, gastric, liver, lung, bladder, breast, ovarian, pancreatic cancer, etc. Genomic analyses have demonstrated that PDXOs largely preserve the histo- and molecular pathology of the original PDX. In vitro maintenance of PDX models as organoids enables genetic manipulation to be performed, which is not possible for in vivo PDX models, thereby creating new models to facilitate drug development. We have employed both lentivirus-based and CRISPR/Cas9-based approaches to modify various tumor organoid models with luciferase, tumor associated antigens (TAA), and some EGFR mutants unavailable in current preclinical model panels. These engineered organoid models, alongside corresponding in vivo matched models, have been incorporated into a new drug efficacy platform. Preliminary results show that these engineered organoid models greatly expand the capacity and application of our current organoid bank. Conclusions: In summary, engineered cancer organoids are a great tool to enrich and diversify current tumor organoid biobanks, particularly for unavailable and clinically relevant models to be used for drug testing, as well as for studies on mechanisms of action and drug resistance.

Citation Format: Fei Wang, Rongyun Zhou, Xinlong Dong, Yuqing Ma, Jun Zhou, Xiaoxi Xu, Yujun Huang, Henry Li, Phillip S-Z Wang. The emergence of tumor organoid engineering as a powerful tool for drug efficacy and mechanism of action profiling [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 LB-007.
Abstract LB-018: Chemotherapeutic stress influences epithelial-mesenchymal transition and stemness in cancer stem cells of triple-negative breast cancer
Authors: Li Xiao , Strietz Juliane , Bleilevens Andreas , Stickeler Elmar , Maurer Jochen
Published: September 2020

Triple-negative breast cancer (TNBC) is a subtype of breast cancer characterized by the absence of estrogen and progesterone receptors (ER, PR) and lacking an overexpression of human epidermal growth factor receptor 2 (HER2). Apart from this lack of therapeutic targets, TNBC also shows an increased capacity for early metastasis and therapy resistance. Currently, many TNBC patients receive neoadjuvant chemotherapy (NACT) upon detection of the disease. With TNBC likely being driven at least in part by a cancer stem-like cell type, we wanted to evaluate the response of primary cancer stem cells (CSCs) to standard chemotherapeutics. Therefore, we set up a survival model using primary CSCs to mimic tumor cells in patients under chemotherapy. Breast cancer stem cells (BCSCs) were exposed to chemotherapeutics with a sublethal dose for six days. Surviving cells were allowed to recover in culture medium without chemotherapeutics. Surviving and recovered cells were examined in regard to proliferation, migratory capacity, sphere forming capacity, epithelial-mesenchymal transition (EMT) factor expression at the mRNA level, and cancer-related microRNA (miRNA) profile. Our results indicate that chemotherapeutic stress enhanced sphere forming capacity of BCSCs, and changed cell morphology and EMT-related gene expression at the mRNA level, whereas the migratory capacity was unaffected. Six miRNAs were identified as potential regulators in this process.

Citation Format: Xiao Li, Juliane Strietz, Andreas Bleilevens, Elmar Stickeler, Jochen Maurer. Chemotherapeutic stress influences epithelial-mesenchymal transition and stemness in cancer stem cells of triple-negative breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr LB-018.
Abstract LB-020: The replication factor MCM10 maintains breast cancer stem-like cells that constitutively experience c-Myc-driven replication stress
Authors: Gotoh Noriko
Published: September 2020

Cancer stem-like cells (CSCs) are thought to be responsible for recurrence of tumors and poor prognosis due to their higher tumor-initiating capacity and resistance against chemotherapeutic agents, although the underlying mechanisms remain unclear. Here, we uncover a critical role of the replication factor mini-chromosome maintenance protein 10 (MCM10) in enabling breast cancer-derived CSCs to deal with constitutively accumulating DNA replication stress. Transcriptomic analysis of patient-derived breast cancer cells revealed that c-Myc downstream target genes and replication stress response genes were highly upregulated in CSC-enriched spheroid cells. Upregulated c-Myc enhanced the transcriptional activity especially at nucleoli, leading to DNA replication stress caused by collision of transcription and replication machineries. MCM10, one of the most upregulated factors in CSCs, showed important roles for enabling cells to survive under such condition through activating dormant origins. MCM10 depletion decreased tumorigenesis, tumor sphere-forming capacity, expression levels of stemness markers, and tumor-initiating capacity, suggesting that elevated MCM10 levels are vital for CSC survival and proliferation. On the other hand, overexpression of MCM10 helped cells to proliferate in medium containing hydroxyurea (HU), a chemotherapeutic agent that induces replication stress. We therefore conclude MCM10 plays critical roles for cancer cell proliferation under DNA replication stress, which is more pronounced in CSCs with upregulated c-Myc activity and in cells treated with chemotherapeutic agents, and propose to target MCM10 for therapy of both CSCs and other cancer cells.

Citation Format: Noriko Gotoh. The replication factor MCM10 maintains breast cancer stem-like cells that constitutively experience c-Myc-driven replication stress [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 LB-020.
Abstract LB-055: Novel therapeutic option targeting the tumor cell lysosomes
Authors: Joncour Vadim Le , Filppu Pauliina , Holopainen Minna , Hyvönen Maija , Turunen S. Pauliina , Sihto Harri , Burghardt Isabel , Jääskeläinen Juha , Weller Michael , Lehti Kaisa , Käkela Reijo , Laakkonen Pirjo
Published: September 2020

Malignant brain tumors represent a surgical and therapeutic challenge. Despite major breakthrough in the precision neurosurgery and the personalized medicine, it remains impossible to circumvent the tumor relapse. This is due to the highly infiltrative nature of the glioma cells, with single tumor-initiating cells invading far distances in the brain. However, we recently identified a major flaw in the invasive tumor cell biology: a high sensitivity to the lysosomal membrane permeabilization (LMP). We first identified the fatty acid binding-protein 3 (FABP3) specifically expressed by invasive gliomas cells. Expression of FABP3 is indispensable to the transportation of poly-unsaturated fatty acids (PUFAs), which cannot be otherwise synthetized by the cells, from the extracellular space to biological membranes. Silencing of FABP3 stopped the supply of PUFAs that are essential to the lysosomal membrane maintenance. Induction of the LMP activated a cascade of events including the release of the lysosomal cathepsins in the cytosol, leading to the tumor cell death. Interestingly, the specific killing of invasive glioma cells through the LMP can be achieved by using lysosomotropic drugs, such as anti-histamines. These cationic amphiphilic molecules accumulate in the lysosome and induce the phospholipidosis of the lysosomal membrane. We therefore tested the ability of the blood-brain-barrier permeable clemastine, an anti-histamine, to reduce the tumorigenicity several patient-derived gliomas implanted in nude mice. Although clemastine treatment did not modify the primary tumor mass growth, it completely eradicated the invasive population. These promising results obtained by therapeutic activation of the LMP could also be beneficial for other aggressive cancers models, such as brain metastases of breast cancer or metastatic pancreatic cancer.

Citation Format: Vadim Le Joncour, Pauliina Filppu, Minna Holopainen, Maija Hyvönen, S. Pauliina Turunen, Harri Sihto, Isabel Burghardt, Juha Jääskeläinen, Michael Weller, Kaisa Lehti, Reijo Käkela, Pirjo Laakkonen. Novel therapeutic option targeting the tumor cell lysosomes [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 LB-055.
Abstract LB-130: Suppression of tumor immune activity in adrenocortical carcinoma with excess glucocorticoid
Authors: Greenstein Andrew E. , Habra Mouhammed Amir , Mura Marzena , Biernat Paweł , Piatek Marek J. , Grauer Andreas , Shepherd Stacie Peacock
Published: September 2020

Introduction: Elevated glucocorticoid (GC) activity, while difficult to accurately quantify, has been implicated in the pathophysiology of multiple cancer types. Approximately half of adrenocortical carcinoma (ACC) patients exhibit excess GC (GC+), which provides a unique test case to assess correlates of GC activity. The broad immunosuppressive effects of GC may limit tumor immune response and immune checkpoint inhibitor (ICI) efficacy. ACC multi-omics were analyzed to identify the molecular consequences of GC activity and assess the rationale for combining relacorilant, a glucocorticoid receptor (GR) antagonist, with an ICI in GC+ ACC.

Methods: GC status, mRNA expression, DNA mutation, and DNA methylation data from distinct adrenal resections (n=71) were accessed via The Cancer Genome Atlas (TCGA). To deconvolute immune cell type abundance, xCell was applied to the mRNA data. Random forest was used to derive gene signatures.

Results: The expression of 858 genes differed significantly between GC- and GC+ ACC cases. KEGG pathway analysis showed higher gene expression of 7 pathways involved in steroid synthesis and secretion in GC+ cases. Nineteen pathways showed lower expression, most of which were related to natural killer cells, T-cells, and immune activity. Hypomethylation was primarily observed in the steroid synthesis pathways. Tumor-infiltrating CD4+ memory (P=.003), CD8+ memory (P<.001), and NKT-cells (P=.014) were depleted in GC+ cases, while tumor-associated neutrophils were enriched (P<.001). Higher tumor mutation burden (TMB) was observed in the GC+ cases (P=.029). A gene signature predictive of GC status in ACC was derived and applied to other cancer types to identify tumors with GC+-like transcriptional profiles.

Conclusions: Reduced abundance of immune cells and immune-related transcripts in GC+ ACC provides insight into the mechanisms by which GC may limit response to ICI therapy. GR antagonism may increase immune related transcripts, thus promoting tumor immune response in GC+ ACC and other malignancies with elevated GC activity. This hypothesis will be tested in a Phase 1 trial of relacorilant + ICI.

Citation Format: Andrew E. Greenstein, Mouhammed Amir Habra, Marzena Mura, Paweł Biernat, Marek J. Piatek, Andreas Grauer, Stacie Peacock Shepherd. Suppression of tumor immune activity in adrenocortical carcinoma with excess glucocorticoid [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 LB-130.
Abstract LB-131: Profile of glucose metabolism in bladder cancer patients and the inhibitory effect of sulforaphane on bladder cancer glycolysis by regulating microRNA expression
Authors: Shan Yujuan , Huang Lei
Published: September 2020

Object: We are committed to studying the characteristics of energy metabolism in bladder cancer patients, and to investigate the regulation of sulforaphane (SFN) on glucose metabolism in bladder cancer cells. Methods: Real-time PCR was used to detect the gene level of glycolysis key rate-limiting enzymes in bladder tumors, and para-carcinoma tissues as control. We also measured the activity of glycolysis enzymes in patient serum and bladder cancer cells by ELISA. Then we measured microRNAs level by microarray after SFN treatment. GO function and KEGG pathway enrichment analyses were used to gain insights into the biological roles and the potential pathway of different expression microRNAs. Results: Gene expression results showed that the level of PFK-1 was highly increased in tumor tissues. However, there is no significant differential in serum activity of PFK-1 as compared with control group. SFN obviously elevated microRNA expression related to glucose metabolism, including miR-200c, miR-34a and let-7. The activity of HK2, which can be blocked by miR-34a, was strongly inhibited in bladder cancer cells by incubating with 20 μmol/L SFN for 24 h. Furthermore, functional annotation analysis results suggest that miR-200c target genes were mainly enriched in 82 GO terms and 17 pathways (P < 0.05), including mTOR signaling (p = 0.0154) and AMPK signaling (p = 0.0158), which also involved in the regulation of energy metabolism. Conclusion: Aberrant gene expression of PFK-1 is one of most important causes of glucose metabolic disorders in bladder cancer patients. SFN can significantly down-regulate glucose metabolism in bladder cancer cells by regulating the expression of glycolysis related-microRNAs.

Citation Format: Yujuan Shan, Lei Huang. Profile of glucose metabolism in bladder cancer patients and the inhibitory effect of sulforaphane on bladder cancer glycolysis by regulating microRNA 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 LB-131.
Abstract LB-163: Cost-effectiveness analysis of HBV-related hepatocellular carcinoma screening in cirrhosis patients with antiviral treatment in China
Authors: Zhang Meng , Li Yaoguang , Zhang Shunxiang , Zhang Jianying
Published: September 2020

Background/Aims: The majority of hepatocellular carcinoma (HCC) patients (>80%)are attributable to hepatitis B virus (HBV) in China. Antiviral treatment is a public health strategy to slow down or change the progression of the disease so as to prevent HCC. In recent years, the Chinese government has been reducing the price of antiviral drugs responding to WHO program of global hepatitis B elimination. Therefore, we modelled the consequences of different screening strategies with or without antiviral treatment to evaluate the cost-effectiveness for HBV-related cirrhosis patients in China. Methods: We designed a markov model to compare no screening with 8 screening strategies in HBV-related cirrhosis patients aged 40 and older in China: annual/semiannual US, annual/semiannual US with AFP, annual/semiannual CT and annual/semiannual MRI. Meanwhile, to evaluate the cost-effectiveness of the strategies in the combination of screening and the antiviral treatment, we also assessed the other 8 screening strategies with antiviral treatment compared to the 8 non-antiviral strategies accordingly. Costs, quality-adjusted life-years (QALYs), life-years (LY) and incremental cost-effectiveness ratios (ICERs) were collected and used to assess the cost-effectiveness of screening strategies and antiviral therapeutics. Results: Semiannual CT strategy was less cost and more effective than no screening with 0.81 QALYs gained and $1092 saved. The ICER ($/QALY) for semiannual MRI strategy compared with semiannual CT strategy was $220,318 that was higher than the WTP ($29,331/QALY). As to antiviral treatment strategies, Semiannual CT strategy was also less cost and more effective than no screening with 0.48 QALYs gained and $535 saved. Not only was Semiannual CT the most efficacious strategy, but also the ICERs ($/QALY) were $4,681/QALY, $6,814/QALY and $11,560/QALY as the results by comparing annual US with AFP strategy, annual CT strategy and semiannual US with AFP strategy to semiannual CT strategy respectively. The QALY of semiannual CT with antiviral treatment increased by 5.37 compared to semiannual CT without antiviral treatment. The ICER was $3,944/QALY. Conclusions: Antiviral treatment appears to be a cost-effective strategy in HBV-related cirrhosis populations in China and semiannual CT with antiviral treatment strategy may be more cost-effective for HBV-related hepatocellular carcinoma Screening and prevention.

Citation Format: Meng Zhang, Yaoguang Li, Shunxiang Zhang, Jianying Zhang. Cost-effectiveness analysis of HBV-related hepatocellular carcinoma screening in cirrhosis patients with antiviral treatment in China [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 LB-163.
Abstract LB-183: Sensitivity of cancer cells to oncolytic viruses is defined by IWS1 phosphorylation dependent epigenetic regulation of U2AF2 splicing and nucleocytoplasmic export of type I IFN transcripts
Authors: Laliotis Georgios I. , Kenney Adam D. , Orlacchio Arturo , Coppola Vincenzo , Yount Jacob S. , Tsichlis Philip N.
Published: September 2020

We have previously shown that phosphorylation of IWS1 by AKT (primarily AKT3) at Ser720/Thr721 promotes the binding of SETD2 to an Spt6/IWS1/Aly-Ref transcription elongation complex. This complex is bound via Spt6 to the Ser2-phosphorylated CTD of RNA Pol II. We have also shown that this results in the AKT-dependent trimethylation of Histone H3 at K36, a histone modification that contributes to the regulation of alternative splicing, and that this splicing pathway plays an important role in lung cancer. Here, we show that one of the genes targeted for alternative splicing via this mechanism is U2AF2, which encodes the splicing factor U2AF65. The predominant alternatively spliced U2AF2 transcript in cells which have lost IWS1, and in cells in which IWS1 is not phosphorylated, lacks exon 2, which encodes the SR domain of U2AF65. U2AF65 lacking the SR domain does not bind Prp19, a member of a seven-protein complex, which also contributes to RNA processing. U2AF65 and Prp19, along with the TREX complex, bind Cytoplasmic Accumulation Response Elements (CAR-E) in naturally intronless mRNAs and promotes their nucleocytoplasmic export. Examples of mRNAs whose transport is regulated by this mechanism include IFNα1, IFNβ1, HSPBP3 and c-Jun. Based on these data, we predicted that the production of type I IFNs would be dependent on IWS1 phosphorylation by AKT and that inhibition of this pathway would lead to increased sensitivity of cells to virus infection. Indeed, type I IFN production was decreased in such cells and infection with Vesicular Stomatitis Virus (VSV) was accordingly increased. In addition, whereas U2AF65 binds CAR-E elements in the RNA of type I interferons independent of IWS1 phosphorylation, the binding of Prp19 to the same RNA elements occured only in cells expressing AKT-phosphorylated IWS1. These data identify a mechanism by which AKT inhibits viral infection and suggest that inhibition of the AKT/IWS1 axis will increase the sensitivity of cancer cells to infection by oncolytic viruses.

Citation Format: Georgios I. Laliotis, Adam D. Kenney, Arturo Orlacchio, Vincenzo Coppola, Jacob S. Yount, Philip N. Tsichlis. Sensitivity of cancer cells to oncolytic viruses is defined by IWS1 phosphorylation dependent epigenetic regulation of U2AF2 splicing and nucleocytoplasmic export of type I IFN transcripts [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 LB-183.
Abstract LB-184: A lncRNA-histone acetyltransferase complex induces colorectal oncogenesis through regulation of a metabolic kinase network
Authors: Polytarchou Christos , Hatziapostolou Maria , Koutsioumpa Marina , Christodoulou Niki , Yau Tung On , Birli Eleni , Pomenya Odette , Montiel-Duarte Cristina , Joshi Swapna Mahurkar , Hommes Daniel W. , Verspaget Hein W. , Yu Jun , Iliopoulos Dimitrios
Published: September 2020

Long non-coding RNAs (LncRNAs) through interactions with other cellular macromolecules drive cell processes contributing to important cancer phenotypes. Here we show that HOTAIR, a highly up-regulated lncRNA in human colon cancers, induces transformation of human non-transformed colon epithelial cells. An exon 6 HOTAIR region directly binds the histone acetyltransferase PCAF (KAT2B). The HOTAIR/PCAF complex regulates H3K27 acetylation and transcriptional activation preferentially of genes encoding components of the PI3K/AKT/MEK pathways and induces cell metabolic reprogramming. The transcriptional signature of this HOTAIR/PCAF metabolic circuit is active in colon cancer and correlates with overall patient survival. Topological and mechanistic analyses were employed to characterize the HOTAIR/PCAF interface and design a reversible specific inhibitor (CPDI-1). Selective blocking of the HOTAIR-PCAF interaction, suppresses AKT activation and tumor growth. In conclusion, we demonstrate that disruption of a lncRNA-histone modifier complex interferes with the cancer epigenome and represents a novel approach for cancer therapeutics.

Citation Format: Christos Polytarchou, Maria Hatziapostolou, Marina Koutsioumpa, Niki Christodoulou, Tung On Yau, Eleni Birli, Odette Pomenya, Cristina Montiel-Duarte, Swapna Mahurkar Joshi, Daniel W. Hommes, Hein W. Verspaget, Jun Yu, Dimitrios Iliopoulos. A lncRNA-histone acetyltransferase complex induces colorectal oncogenesis through regulation of a metabolic kinase network [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 LB-184.