bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3911: High-throughput screening of patient-derived HCC organoids for drug discovery
Authors: Wong Alissa Michelle Go , Cheung Winnie , Wong Maggie , Wong Aikha , Lam Helena , Zhang Zhe , Man Matthew , Chen Hui , Close David , Johnston Paul , Chan Anthony , Lai Paul , Kwong Joseph , Wong Nathalie
Published: September 2020

Current in vitro models of primary liver cancer insufficiently recapitulates the tumor heterogeneity and pathophysiology of the original patient's tumor. This significantly hinders translational studies and often leads to the high rate of clinical trial attrition and poor patient response. Recent technological advances in organoid culture methods have been reported to overcome these limitations. Our group has successfully established 3 patient-derived HCC tumor organoids that closely recapitulate the patient's pathophysiology. Conventional short-term monolayer culture of the same 3 tumors as well as 3 non-tumoral adjacent liver and 3 normal liver organoids were established and compared with 5 HCC cell lines from ATCC for their drug response profiles. Subsequent drug screening was performed using a panel of 100 drugs, of which 70 are FDA-approved for either HCC or other cancers, as well as 30 pipeline compounds targeting frequently mutated pathways in HCC. We found that our screening pipeline could achieve robust z' scores and reproducible data points with technical and biological replicates. Principal component analysis of Total Area under curve (AUC) values show distinct and progressive changes between Normal Liver, Non-tumoral adjacent liver, and Tumor organoids. Upon comparison with HCC cell lines Hep3B, HepG2, Huh7, SNU398 and PLC5, we found HCC patient-derived short term cultures, particularly organoids have more diverse drug sensitivity profiles and may be a better indicator of patient heterogeneity. Interestingly, FDA-approved drugs for HCC such as Regorafenib show high sensitivity and tumor specificity to ATCC cell lines (ATCC vs Normal liver P=0.00017***), but this specificity was reduced and lost in patient-derived 2D cultures (P=0.045*) and organoids (P=0.895), respectively. This may explain the observed hepatotoxicity observed in HCC patients and warrant further investigation of more targeted ways of treating HCC.

Citation Format: Alissa Michelle Go Wong, Winnie Cheung, Maggie Wong, Aikha Wong, Helena Lam, Zhe Zhang, Matthew Man, Hui Chen, David Close, Paul Johnston, Anthony Chan, Paul Lai, Joseph Kwong, Nathalie Wong. High-throughput screening of patient-derived HCC organoids for drug discovery [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 3911.
Abstract 394: LXR agonist N,N-dimethyl-3-β-hydroxy-cholenamide, DMHCA, reduces ErbB2-dependent tumorigenesis and immune tolerance in a conditional model of mammary fibrosis
Authors: Glazer Robert I. , Sheng Gao , Yuan Hongyan , Ranjit Suman , Lu Jin , Lu Xun , Levi Moshe
Published: September 2020

Background: One of the central challenges for cancer therapy is the identification of factors in the tumor microenvironment that increase tumor progression and prevent the immune system from eradicating the tumor. One such factor associated with breast cancer is stromal fibrosis, a histopathologic criterion for invasive cancer and poor survival. Fibrosis is caused by inflammatory factors and remodeling of the extracellular matrix, which results in excessive collagen deposition and an immune tolerant environment. Experimental Procedures: We developed a novel transgenic model representative of HER2/ErbB2+ breast cancer, termed NeuT/ATTAC, in which partial ablation of mammary fat resulted in its replacement with fibrotic tissue. NeuT/ATTAC mice were administered a diet supplemented with 0.04% DMHCA, a desmosterol-like pan agonist of the liver X receptor (LXR). Collagen deposition resulting from fibrosis was assessed by second harmonic generation (SHG) fluorescence microscopy and histological staining, immune cell subsets were analyzed by fluorescence-activated cell sorting and biomarkers of fibrosis, proliferation and immune tolerance were determined by IHC. Findings: Administration of DMHCA to NeuT/ATTAC mice beginning at two months of age reduced tumorigenesis, tumor multiplicity and suppressed fibrosis as shown by loss of collagen, fibroblast activation protein (FAP) and α-smooth muscle actin (SMA). These changes were accompanied by a marked reduction in the percentage of MDSC, and an increase in CD8+ effector T cells and PD1+/Foxp3+ CD4 T cells.

Conclusion: Reduction of fibrosis, tumorigenesis and immune tolerance by DMHCA suggests that this LXR agonist may be useful for reducing tumor-associated fibrosis and in combination with immune checkpoint inhibitors that target PD1+ Treg cells for moderating immune tolerance. Unlike non-steroidal LXR agonists, DMHCA does not increase serum triglycerides or produce hepatic steatosis, and may therefore be a good candidate for clinical studies. Supported by a Sher Award, 1P30CA051008 from the NCI, NIH, to the Lombardi Comprehensive Cancer Center, and R01 HL133545 and R01 DK116567 from the NIH to M.L.

Citation Format: Robert I. Glazer, Gao Sheng, Hongyan Yuan, Suman Ranjit, Jin Lu, Xun Lu, Moshe Levi. LXR agonist N,N-dimethyl-3-β-hydroxy-cholenamide, DMHCA, reduces ErbB2-dependent tumorigenesis and immune tolerance in a conditional model of mammary fibrosis [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 394.
Abstract 3965: Novel biomimetic, interfacing 3D hydrogels to investigate roles of glioblastoma (GBM) niche on GBM progression
Authors: Sohrabi Alireza , Liang Jesse , Safarians Gevick , Guivatchian Elnaz , Solomon Itay , Seidlits Stephanie
Published: September 2020

Glioblastoma (GBM) is the most common and lethal form of brain tumors with survival rate of 5% after 5 years. In the tumor microenvironment, invading GBM cells experience various ECM transitions at the tumor margin, such as alterations in matrix stiffness. Although conventional three-dimensional (3D) hydrogels have improved upon existing in vitro tumor models, these biomaterial platforms are not capable of separating the migratory GBM population from the bulk of the tumor. In addition, conventional hydrogels can only mimic a single microenvironment property (i.e. stiffness and protein presentation). In this study, we fabricated interfacing hyaluronic acid (HA)-based hydrogels using thiol-ene photochemistry to mimic the distinct stiffness conditions of tumor and surrounding tissue as well as the stiffness transition. The novel interfacing model enabled us to isolate invading GBM cells for further in vitro analysis. Atomic Force Microscopy (AFM) analysis of our xenograft GBM tumors in mice brain demonstrated that GBM tumor is stiffer than surrounding brain tissue. Therefore, mechanical properties of each hydrogel condition were characterized via AFM to confirm similarity of hydrogel stiffness to GBM xenografts and patient-derived, primary GBM cell migration in the HA-hydrogels was investigated. Immunohistochemistry staining showed that invading GBM cells, passing the interface have higher expression of RHAMM and CD44. Our findings suggest that invading GBM cells might have different expression profile and therefore require more thorough analyses using single cell RNA-seq.

Citation Format: Alireza Sohrabi, Jesse Liang, Gevick Safarians, Elnaz Guivatchian, Itay Solomon, Stephanie Seidlits. Novel biomimetic, interfacing 3D hydrogels to investigate roles of glioblastoma (GBM) niche on GBM 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 3965.
Abstract 3977: Stromal BMP signaling imbalance mediated by GREM1 and ISLR regulates colorectal cancer progression
Authors: Kobayashi Hiroki , Gieniec Krystyna A. , Wang Tongtong , Wright Josephine A. , Suzuki Nobumi , Lannagan Tamsin RM , Hayakawa Yoku , Leedham Simon J. , Arpaia Nicholas , Mukherjee Siddhartha , Wang Timothy C. , Enomoto Atsushi , Takahashi Masahide , Worthley Daniel L. , Woods Susan L.
Published: September 2020

Introduction

Cancer-associated fibroblasts (CAFs), a heterogeneous component of the tumor microenvironment, substantially influence tumor progression. Bone morphogenetic proteins (BMP) play a critical part in defining the intestinal epithelial niche and either promote or retard cancer progression in a context-dependent manner. However, the role of BMP signaling in the colorectal cancer (CRC) stroma remains to be fully elucidated. This study investigated the significance of mesenchymal BMP signaling as a potential therapeutic target in CRC progression.

Design

Using CRC expression array data, we identified two CAF-specific factors involved in BMP signaling, then verified their upregulation in the human CRC stroma by in-situ hybridization (ISH). We took advantage of a preclinical mouse model of CRC hepatic metastasis to test approaches targeting the BMP signaling pathway.

Results

CRC microarray data identified GREM1 and ISLR as CAF-specific genes involved in BMP signaling. In colonic myofibroblasts, Grem1-overexpression inhibited BMP signaling whereas BMP7 signaling was augmented by Islr overexpression, suggesting opposing roles for GREM1 and ISLR in the regulation of BMP signaling. ISH using human rectal cancer samples revealed that GREM1 and ISLR were expressed in distinct CAF subpopulations and that GREM1 and ISLR expression predicted poor and favorable survival, respectively. Notably, Grem1 and Islr expression was differentially regulated by Foxl1, an intestinal mesenchyme-lineage transcription factor, and TGF-b, indicating a mechanism for generating fibroblast heterogeneity. Finally, adeno-associated virus 8-mediated in-vivo overexpression of Islr in hepatocytes retarded growth and generated more differentiated histology in CRC hepatic metastases.

Conclusion

These data suggest that increased stromal BMP signaling may ameliorate CRC progression and provide a rationale for targeting stromal BMP signaling to inhibit CRC progression and metastasis.

Citation Format: Hiroki Kobayashi, Krystyna A. Gieniec, Tongtong Wang, Josephine A. Wright, Nobumi Suzuki, Tamsin RM Lannagan, Yoku Hayakawa, Simon J. Leedham, Nicholas Arpaia, Siddhartha Mukherjee, Timothy C. Wang, Atsushi Enomoto, Masahide Takahashi, Daniel L. Worthley, Susan L. Woods. Stromal BMP signaling imbalance mediated by GREM1 and ISLR regulates colorectal 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 3977.
Abstract 4002: Adipose tissue supernatant of overweight ER positive breast cancer patients promotes migration of MCF-7 breast cancer cells
Authors: Ma Jie , Timmer-Bosscha Hetty , Werker Paul M. , Schröder Carolina P. , Arjaans Marlous
Published: September 2020

Introduction: Adipose tissue is a major component of the breast cancer microenvironment, and preclinical studies have suggested that adipose tissue might play a pivotal role in breast cancer progression. In this study we investigated the effect of adipose tissue from patients with ER positive breast cancer on migration of ER positive MCF-7 breast cancer cells.

Materials and Methods: Adipose tissue samples were collected from patients with ER positive breast cancer undergoing a mastectomy. In total, adipose tissue samples from 3 normal weight (BMI: 18.5-24.9) and 7 overweight breast cancer patients (BMI: 25-29.9) were collected. Samples from the mastectomy specimens were minced in small pieces, incubated in RPMI-1640 and after 48 hours (h) the supernatant was harvested. The effect of adipose tissue supernatant on the migration of MCF-7 cells was monitored in real-time for 48 h with the xCELLigence cell invasion and migration plates expressed in delta cell index (DCI). In addition, effect of adipose tissue supernatant on migration was assessed by quantification of F-actin positive lamellipodia of MCF-7 cells in a scratch assay after 24 h. In all experiments, RPMI-1640 with 1% or 20% FCS was used as a negative or positive control respectively.

Results: Supernatant of overweight patients significantly induced MCF-7 migration compared to supernatant of normal weight patients after 24 h (xCELLigence DCI: 1.22 ± 0.15 vs. 0.95 ± 0.19 (P=0.038)), 36 h (DCI 1.40 ± 0.21 vs. 0.91 ± 0.31 (P=0.019)) and 48 h (DCI 1.72 ± 0.23 vs. 0.98 ± 0.43 (P=0.007)). In addition, there was a significant difference between the overweight group and negative control at 36 h, and 48 h (DCI 1.40 ± 0.21 vs. 1.06 ± 0.12, P=0.038 and DCI 1.72 ± 0.23 vs. 1.11 ± 0.21, P=0.004 respectively). No significant difference between normal weight group and negative control was found at any of the time points. After 24 h MCF-7 cells incubated with supernatant of overweight patients developed significantly more F-actin positive lamellipodia compared to MCF-7 cells incubated with supernatant of normal weight patients (44.26 ± 3.26% vs. 25.47 ± 1.88% P<0.001).

Conclusion: In this study, we showed that adipose tissue supernatant of patients with ER positive breast cancer can promote migration of MCF-7 breast cancer cells in a weight dependent manner. Whether this is the case in other breast cancer subtypes as well, and the potential clinical implication, is of interest for future studies.

Citation Format: Jie Ma, Hetty Timmer-Bosscha, Paul M. Werker, Carolina P. Schröder, Marlous Arjaans. Adipose tissue supernatant of overweight ER positive breast cancer patients promotes migration of MCF-7 breast cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4002.
Abstract 4014: Measuring direct binding to epigenetic targets using SPR
Authors: Eells Rebecca , Baker Lynn K. , Bray Christopher , Ma Haiching
Published: September 2020

Epigenetic modifications are dynamic and reversible processes that regulate gene expression without altering the underlying sequence of the DNA. Gene expression is instead controlled by chromatin modifications, such a DNA methylation, histone modification, and chromatin remodeling. Proteins involved in epigenetic modifications are categorized as writers, which modify the chromatin; readers, that recognize the modifications; and erasers which remove the modifications. While essential for cellular function, abnormal expression or alteration of the proteins involved in epigenetic regulation leads to diseases, such as cancer. Due to their role in disease, epigenetic modifiers are attractive targets for the discovery and development of novel therapeutics. Research in the field has shown many epigenetic proteins are druggable and several FDA-approved drugs have already been developed (Ref. 1). At Reaction Biology we offer a suite of services for epigenetic drug discovery, including the largest panel for epigenetic screening and profiling in the industry with over 30 Methyltransferases, 100+ Reader Domain Proteins, HATs, HDACs, Demethylases, DUBs, PARPs, and more. In order to directly probe the strength of the binding interaction between the epigenetic target and potential therapeutics we offer biophysical assays, such as surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), and microscale thermophoresis (MST). Here we present SPR binding data for three epigenetic targets: NSD2 (writer), PRMT5 (writer), and Bromodomain-4 (reader). Through the use of commercially available controls, we developed SPR assays for these targets that can be used to screen compound or fragment libraries, determine kinetics/affinity, and profile binding specificity. (Ref 1.) V. Prachayasittikul et al (2017) Exploring the epigenetic drug discovery landscape. Expert Opin. Drug Discov. 12(4): 345-362.

Citation Format: Rebecca Eells, Lynn K. Baker, Christopher Bray, Haiching Ma. Measuring direct binding to epigenetic targets using SPR [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 4014.
Abstract 4037: In vitro cytotoxicity of CPX-351 plus venetoclax compared with hypomethylating agents or cytarabine plus venetoclax
Authors: Gupta Aparna , Humphreys Robin , Hanson Derek , Wang Qi
Published: September 2020

Introduction: Current treatment options for AML patients who are older or unfit for intensive chemotherapy are limited and associated with less than ideal outcomes, even with newly approved regimens of venetoclax plus low-dose cytarabine or DNA hypomethylating agents. CPX-351, a dual-drug liposomal encapsulation of cytarabine and daunorubicin at a synergistic ratio, was approved by the US FDA in 2017 and EMA in 2018 for treatment of adults with newly diagnosed therapy-related AML and AML with myelodysplasia-related changes. The objective of this study was to assess the cytotoxic effects of CPX-351 plus venetoclax versus combinations of venetoclax plus cytarabine, decitabine, or azacitidine.

Methods: Three AML and three ALL cell lines were treated with single-agent CPX-351, venetoclax, cytarabine, decitabine, or azacitidine, or with combinations of CPX-351 plus venetoclax, venetoclax plus cytarabine, venetoclax plus decitabine, or venetoclax plus azacitidine. The cells were plated 24 hours prior to treatment and incubated with the test agents for 72 hours. Cytotoxicity was measured with a luminescent Cell Viability Assay.

Results: As a single agent, all tested drugs demonstrated cytotoxicity to all cell lines, but with different potencies. CPX-351 plus venetoclax showed synergistic or additive effects in all AML and ALL cell lines tested (Table). Combinations of venetoclax with cytarabine, decitabine, or azacitidine showed variable responses (synergistic, additive, and antagonistic effects). In addition, data suggest the highest combination doses with CPX-351 may attain greater cell killing versus the other drug combinations tested.

TableCICell lineCell originCPX-351 + venetoclaxCytarabine + venetoclaxAzacitidine + venetoclaxDecitabine + venetoclaxHL-60AML0.701.010.711.66SKM-1AML0.911.931.291.14KG-1AML0.530.950.590.77CCRF-CEMALL0.700.990.861.99MOLT-3ALL0.740.680.630.73Jurkat E6-1ALL0.791.331.141.40CI was calculated using the Chou and Talalay method, where CI of <1 = synergistic, 1 = additive, and >1 = antagonistic.

Conclusions: These data demonstrated that the combination of CPX-351 and venetoclax is synergistic or additive in all cell lines tested. Future studies confirming these findings in in vivo hematologic malignancy models can help to design a novel dose-regimen, which may provide the best benefit with good tolerability.

Citation Format: Aparna Gupta, Robin Humphreys, Derek Hanson, Qi Wang. In vitro cytotoxicity of CPX-351 plus venetoclax compared with hypomethylating agents or cytarabine plus venetoclax [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 4037.
Abstract 4051: Anlotinib inhibits the progress of colorectal cancer cells by antagonizing glycolysis & JAK2-STAT3 signaling
Authors: Jia Zhenxian , Zhang Zhi , Li Ze , Li Ang , Xie Yuning , Wu Hongjiao , Yang Zhenbang , Zhang Hongmei , Zhang Xuemei
Published: September 2020

Background: Anlotinib is a novel multi-target tyrosine kinase receptor inhibitor (TKI) that has been proved to treat advanced NSCLC as a third-line regime in China. This study designed to investigate the effectiveness & possible mechanism of anlotinib colorectal cancer (CRC) therapy.

Methods: CCK-8 assay & CyQUANT NF assay were used to detect the sensitivity of CRC cells to DDP & anlotinib. Real time quantitative polymerase chain reaction was used to detect the mRNA level of hub genes. The protein expression of pVEGFR2, cleaved PARP, cleaved caspase 3, JAK2, pJAK2, STAT3 & PSTAT3 were determined by western blot. The roles of DDP & anlotinib in the progression, invasion & metastasis of CRC cells were evaluated by colony formation, wound healing & trans-well assay. Lactate release & glucose consumption were measured to investigate glucose metabolism. Cell apoptosis was analyzed by Hoechst33342 fluorescence staining & flow cytometer.

Results: We found that anlotinib may improve the treatment of CRC by blocking the effects of VEGFA. Subsequently, in vitro assays demonstrated that anlotinib &/or DDP inhibited cell proliferation, migration & invasion & also activated apoptosis in human colorectal cancer cells. Additionally, glucose metabolism in CRC HCT-116 & LOVO cells was suppressed by anlotinib &/or DDP. In other hand, anlotinib &/or DDP may improve the treatment of CRC by inhibiting JAK2-STAT3 signaling pathway.

Conclusions: Our preclinical studies showed that anlotinib inhibited cell proliferation, invasion, migration & activated apoptosis by antagonizing glycolysis & JAK2-STAT3 signaling. Combination chemotherapy of anlotinib with cisplatin is more sensitive to CRC.

Citation Format: Zhenxian Jia, Zhi Zhang, Ze Li, Ang Li, Yuning Xie, Hongjiao Wu, Zhenbang Yang, Hongmei Zhang, Xuemei Zhang. Anlotinib inhibits the progress of colorectal cancer cells by antagonizing glycolysis & JAK2-STAT3 signaling [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 4051.
Abstract 4053: Mucin 5ac as a potential therapeutic target for KRAS-driven lung adenocarcinoma
Authors: Velasco Torrez Walter V. , Deng Shanshan , Ramos-Castaneda Marco A. , Del Aguila-Soto Segundo W. , Moghaddam Seyed
Published: September 2020

Lung cancer continues to be the leading cause of cancer death in the United States, with more than 140,000 deaths per year. One of the main types of lung cancer is non-small cell lung cancer with lung adenocarcinoma (LUAD) being the most prominent subtype and is mainly due to oncogenic mutations like KRAS, which offers limited and unsuccessful treatments. The mucus barrier produced by mucins, particularly Mucin 5ac (Muc5ac) which is mainly expressed by lung epithelial cells, is essential for airway homeostasis. However, its overproduction can promote respiratory diseases such as chronic obstructive pulmonary disease (COPD), which is known to increase the risk of lung cancer by 3-10 fold. We have previously shown that Muc5ac is commonly expressed in patients with KRAS-driven LUAD and is associated with poor prognosis. Additionally, we found that lack of Muc5ac (genetic deletion) in mice with LUAD either induced chemically by urethane or genetically engineered with mutant KRAS presented with significantly reduced tumor burden. Therefore, we sought a translational approach by inhibiting Muc5ac in our KRAS-driven lung cancer mutant mouse model using an antisense oligonucleotide (ASO) against Muc5ac and hypothesized that the administration of this drug would result in decreased Muc5ac mRNA expression and reduced tumor burden. This was tested by intrapharyngeal administration of either ASO (5mg/kg) or vehicle (saline) twice a week to a cohort of six-week-old KRASG12D mutant mice for a total duration of eight weeks. Our results showed a 5-fold decrease in Muc5ac mRNA expression and a 42% reduction in overall tumor burden when compared to age and sex matched control mice. Additionally, analysis of bronchoalveolar lavage fluid showed a predominance of macrophages with significant downregulation of the protumorigenic M2 type macrophage polarization markers Fizz1 and Arg1 as well as significant upregulation of TNF-a, which suggests a mechanism mediated by direct tumor cytotoxic activity or apoptosis associated with a reduction in macrophage mediated immunosuppressive tumor microenvironment. In conclusion, our study suggests that administration of Muc5ac ASO effectively inhibits mucin production and ameliorates tumor progression, which ultimately could provide insight in both treatments and prevention against KRAS-driven LUAD.

Citation Format: Walter V. Velasco Torrez, Shanshan Deng, Marco A. Ramos-Castaneda, Segundo W. Del Aguila-Soto, Seyed Moghaddam. Mucin 5ac as a potential therapeutic target for KRAS-driven lung 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 4053.
Abstract 4087: ALDH1A1 inhibitor induces cell cycle arrest and reduces cell proliferation of cisplatin-resistant ovarian cancer cells
Authors: Ramachandran Ilangovan , Ramadoss Sivakumar , Karuppaiyah Selvendiran , Nathan Lauren , Kumaran R. Ileng , Chaudhuri Gautam
Published: September 2020

Introduction: Ovarian cancer is one of the most common malignancies among women that causes severe mortality in USA and globally. Despite advances in cancer treatments in the last few decades, there is no substantial improvement in the survival rates of patients with advanced ovarian cancers. This is mainly attributed to drug resistance and tumor recurrence. Most patients develop resistance to platinum compounds, which are often used as the first-line therapy. Several studies have shown that aldehyde dehydrogenase (ALDH1A1) plays an important role in cancer stemness and confers resistance to cell death induced by platinum compounds used to treat ovarian cancers. Therefore, in this study, we determined the effect of a small-molecule inhibitor of ALDH1A1 (A37) in cisplatin-resistant ovarian cancer cells and assessed it's molecular mechanism of action.

Experimental methods: The cisplatin-resistant ovarian cancer cells (A2780-CR) were exposed to various concentrations of ALDH1A1 inhibitor (A37) and then the cell proliferation by water-soluble tetrazolium salt (WST-1) assay, cell cycle analysis by flow cytometry and oxygen consumption rate (OCR) by Seahorse extracellular flux analysis (Seahorse XFe24 Analyzer) were performed. Furthermore, the protein expression of critical signaling molecules that modulate the proliferation of ovarian cancer cells was assessed by western blot analysis.

Results: A37 treatment drastically decreased the proliferation of A2780-CR cells at 24 and 48 h. In addition, cell cycle analysis revealed a significant increase in G1 phase and dramatic decrease in S phase of the cell cycle upon A37 treatment. Interestingly, A37 suppressed the basal OCR, an indicator of mitochondrial respiration. Furthermore, A37 reduced the protein expression of mammalian target of rapamycin (mTOR), which plays a critical role in cancer cell cycle and proliferation.

Conclusions: Our findings indicate that the ALDH1A1 inhibitor (A37) induces cell cycle arrest and suppresses the cell proliferation of cisplatin-resistant ovarian cancer cells, possibly through downregulation of mTOR signaling. Most importantly, A37 affects the cancer cell metabolism by reducing the mitochondrial respiration. Taken together, our study highlights the importance of potential therapeutic implication of ALDH1A1 inhibitor towards the treatment of chemoresistant-ovarian cancers.

Citation Format: Ilangovan Ramachandran, Sivakumar Ramadoss, Selvendiran Karuppaiyah, Lauren Nathan, R. Ileng Kumaran, Gautam Chaudhuri. ALDH1A1 inhibitor induces cell cycle arrest and reduces cell proliferation of cisplatin-resistant ovarian cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4087.
Abstract 4107: The impact of NF-κB inhibition on the sensitivity of breast cancer cells to chemotherapy-induced apoptosis
Authors: Abdin Shifaa M. , Tolba Mai F. , Zaher Dana M. , Omar Hany A.
Published: September 2020

Aim: The emergence of breast cancer resistance to chemotherapy represents a major obstacle in cancer therapy. Multiple cellular signalling pathways augment such resistance including the nuclear factor (NF)-κB. In the presented work, we investigated the impact of NF-κB inhibition on the sensitivity of breast cancer cells to chemotherapy-induced apoptotic cell death via Annexin V/PI flow cytometry and other molecular biology-based assays.

Methods: Doxorubicin (DOX)-resistant MCF-7 and MDA-MB-231 breast cancer cell lines were developed and utilized for the current study. Cytotoxicity assays were performed to evaluate the median inhibitory concentration (IC50) of different NF-κB pharmacological inhibitors. Chemotherapy-induced apoptosis in parental and resistant breast cancer cells was assessed in the presence or absence of different NF-κB pharmacologic inhibitors such as pentoxifylline (PTX) and bortezomib (BTZ).

Results: Treatment of MDA-MB-231 cell line with a sub-cytotoxic concentration of PTX significantly enhanced DOX-induced apoptosis by 6-fold (p<0.001) in wild type cells, and by 8-fold (p<0.01) in the resistant subclones. Similarly, BTZ significantly boosted the percentage of apoptotic cells by 6-fold (p<0.001) in parental cells co-treated with DOX. However, the same treatment boosted the percentage of apoptotic cells by 11-fold (p<0.01) in DOX-resistant MDA-MB-231 cells. It is noteworthy that the increased sensitivity to DOX-induced apoptosis that was mediated via NF-κB inhibition was more prominent in the resistant clones.

Conclusion: The data generated from this study support the potential merit of NF-κB inhibitors as adjuvants to improve breast cancer sensitivity to chemotherapy. Further mechanistic investigations are currently in progress in our laboratories to characterize the relationship between NF-κB inhibition and key signaling pathways for breast cancer chemoresistance.

Funding: This work was supported by grant # 1801110326-P from the Research Funding Department, University of Sharjah, UAE, and grant #AJF2018050 from Al Jalila Foundation, UAE.

Citation Format: Shifaa M. Abdin, Mai F. Tolba, Dana M. Zaher, Hany A. Omar. The impact of NF-κB inhibition on the sensitivity of breast cancer cells to chemotherapy-induced 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 4107.
Abstract 4121: ORIC-101 overcomes resistance to diverse chemotherapeutics across cancer types
Authors: Zhou Haiying , Barkund Shravani , Pankov Aleksandr , Hegde Ganapati , Kong Wayne , Narayanan Padmini , Sun Jessica D. , Kabbarah Omar , Friedman Lori S. , Daemen Anneleen
Published: September 2020

Preclinical studies have shown that GR activation leads to decreased response to antimetabolites, taxanes, and platinum agents, while GR inhibition enhances therapeutic efficacy. The novel, selective GR antagonist ORIC-101 is under clinical evaluation in combination with anticancer therapies. In this study we set out to assess which cancer types and chemotherapeutics are responsive to combination with ORIC-101. Specifically, we employed caspase assays, in vitro and in vivo tumor growth inhibition studies, and transcriptional profiling in a panel of preclinical models spanning triple-negative breast, ovarian, non-small cell lung cancers, pancreatic ductal adenocarcinoma, hepatocellular carcinoma, sarcoma, and renal cell carcinoma. We found that ORIC-101 reverses GR-mediated antiapoptosis effects towards diverse chemotherapeutics, including paclitaxel, gemcitabine, and cisplatin, across the seven cancer types. Furthermore, ORIC-101 completely overcomes GR-driven chemoprotection and tumor growth in colony formation assays in vitro and in xenograft studies in vivo. At the molecular level, transcriptional profiling and pathway enrichment analysis showed that ORIC-101 fully reverses GR-activated pathways that are directly involved in drug resistance, such as epithelial-to-mesenchymal transition and antiapoptosis, supporting a pan-cancer role of GR as a mediator of therapy resistance. Altogether, ORIC-101, a selective and potent GR antagonist, overcomes resistance to common chemotherapeutics across multiple cancer models. Clinical evaluation of ORIC-101 in combination with nab-paclitaxel is currently ongoing in advanced solid tumors.

Citation Format: Haiying Zhou, Shravani Barkund, Aleksandr Pankov, Ganapati Hegde, Wayne Kong, Padmini Narayanan, Jessica D. Sun, Omar Kabbarah, Lori S. Friedman, Anneleen Daemen. ORIC-101 overcomes resistance to diverse chemotherapeutics across cancer types [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 4121.
Abstract 4174: Anticancer activity of a novel small molecule krct-1 through inhibition of Trk-A signaling in breast cancer cells
Authors: Kim Ah-Young , Kim Jae-Sung
Published: September 2020

The small molecule krct-1 was synthesized as a structurally novel compound. Krct-1 inhibited cell proliferation and tumor growth of breast cancer. In addition, Krct-1 had a capacity to induce activation of several apoptotic markers and downregulate anti-apoptotic markers in various breast cancer cell lines. Furthermore, Krct-1 increased the radiosensitivity of breast cancer cells by promoting DNA damage responses. By performing the kinase profile analysis, we found that Krct-1 inhibited tropomyosin receptor kinase A (Trk-A) and further demonstrated inhibition of its downstream signaling such as ERK and STAT3 in breast cancer cells. Thus, our results suggest that a novel compound Krct-1 have an antitumor activity through Trk-A regulation in breast cancer.

Citation Format: Ah-Young Kim, Jae-Sung Kim. Anticancer activity of a novel small molecule krct-1 through inhibition of Trk-A signaling in breast cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4174.
Abstract 4191: Characterization of a novel anti-glucose transporter inhibitor as a pan-anticancer therapeutic in vitro and in vivo
Authors: Shriwas Pratik , Bergmeier Stephen , Chen Xiaozhuo
Published: September 2020

Cancer has surpassed cardiovascular diseases as the top killer in many US counties. Opportunistic uptake of extracellular nutrients has recently been named as an emerging hallmark of cancer metabolism. Glucose is a key nutrient taken in by cancer cells for satisfying their high energy and biosynthesis needs. Drastic upregulation of glucose uptake and metabolism, a phenomenon called the Warburg effect, is prevalent among cancers. Cancer cells overexpress specific glucose transporters (GLUTS), particularly GLUT1, on their cell surface for uptaking glucose via facilitated diffusion. Considering the addiction of cancer cells to glucose and their sensitivity and vulnerability to changes in glucose supply, Gluts, have been recognized as an attractive anticancer target. Previously, our first-generation GLUT-inhibitor, WZB117, had been characterized in vitro and in vivo as an anticancer therapeutic. In this study, DRB18, a novel and proprietary lead inhibitor of glucose transporters, is characterized in vitro and in vivo by bioassays. DRB18 inhibits growth and proliferation in about 90% of the 60 different cancer cell lines in 9 major cancer types. It shows IC50 values in high nM range in some cell lines in several different cancer types. DRB18 treatment leads to significant changes in glucose-based metabolism in A549 lung cancer cells, including cell death via ER/oxidative stress and necrosis. The docking and Seahorse studies reveal the surprising contributions of macropinocytosis in DRB18 internalization, ultimately leading to better GLUT1 binding and increase in potency in killing cancer cells. DRB18 (10mg/kg) treatment thrice a week significantly reduces growth in Xenograft A549 tumors. Comprehensive in vitro and in vivo metabolomics analysis would be presented to better describe anti-glucose metabolism of activity of DRB18. Further, GLUT1-KO A549 (A549glut1ko) cell lines, in which Glut1 gene has been knocked out, were generated using CRISPR-Cas9 system for further studying GLUT1's roles in A549 cells. Immunoblotting analysis revealed the upregulation of other GLUTs after GLUT1 is knocked out, strongly suggesting the need for a pan-GLUT inhibitor for better inhibition of tumor growth.

Citation Format: Pratik Shriwas, Stephen Bergmeier, Xiaozhuo Chen. Characterization of a novel anti-glucose transporter inhibitor as a pan-anticancer therapeutic in vitro and in vivo [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4191.
Abstract 4198: Role of receptor tyrosine kinase AXL in mediating DNA damage response
Authors: Yeo Xun Hui , Tam Wai Leong , Huang Ruby Yun-Ju
Published: September 2020

AXL is a receptor tyrosine kinase that is often overexpressed in many cancers. It contributes to tumor progression, metastasis and drug resistance through activating downstream signaling cascades, making it an emerging therapeutic target. Moreover, the first-in-class AXL inhibitor R428 (BGB321) was lately approved by the FDA for the treatment of relapsed or refractory acute myeloid leukemia. Previously, R428 (BGB321) was also reported to show selective sensitivity towards ovarian cancers with a Mesenchymal molecular subtype. Recently, a novel role of AXL in the regulation of DNA damage responses has been brought to light. Here, we further explored AXL's role in mediating DNA damage responses by using ovarian cancer as a disease model. AXL inhibition using R428 resulted in the increase DNA damage foci in ovarian cancer cells SKOV3 and HeyA8. This occurred concurrently with the upregulation of classic DNA damage response signaling molecules such as γH2Ax, ATM and ATR. RNA sequencing results further validated this observation, that R428-treated cells showed downregulation of genes involved in cell cycle control and DNA replication, suggesting the decrease in proliferation. Through SILAC co-immunoprecipitation mass spectrometry, we uncovered a novel binding partner of AXL that could potentially regulate the DNA damage repair upon the inhibition of AXL, giving us insights to how AXL inhibition might contribute to DNA damage. Furthermore, our results showed that AXL inhibition rendered cells more sensitive to the inhibition of ATR, a crucial mediator for replication stress, paving ways to the rationale for potential combinatory use of AXL and DNA damage repair inhibitors.

Citation Format: Xun Hui Yeo, Wai Leong Tam, Ruby Yun-Ju Huang. Role of receptor tyrosine kinase AXL in mediating DNA damage response [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 4198.
Abstract 4223: Combination therapy of BCR-ABL-positive B cell acute lymphoblastic leukemia by tyrosine kinase inhibitor dasatinib and c-Jun N-terminal kinase inhibition
Authors: Xiao Xinhua , Jiao Bo , Ren Ruibao
Published: September 2020

The Philadelphia chromosome (Ph), which leads to creation and expression of the fusion gene product BCR-ABL, underlines the pathogenesis of chronic myelogenous leukemia (CML) and a fraction of adult and pediatric acute B-lymphoblastic leukemia (B-ALL). The BCR-ABL tyrosine kinase inhibitors (TKIs) has shown a remarkable clinical activity in patients with CML, but their efficacy in treating Ph+ B-ALL is limited. Identifying additional therapeutic targets is important for the effective treatment of Ph+ B-ALL. In this study, we found that the c-Jun N-terminal kinase (JNK) signaling pathway is abnormally activated in both human and mouse BCR-ABL+ B-ALL cells, but the BCR-ABL TKI does not inhibit JNK activation in these cells. Inhibition of JNK, either by RNAi-mediated downregulation or by JNK inhibitors, could significantly reduce viability of Ph+ B-ALL cells. JNK inhibition by RNAi-mediated downregulation or JNK inhibitors also showed a synergistic effect with the BCR-ABL TKI, dasatinib, in killing Ph+ B-ALL cells in vitro. Furthermore, a potent JNK inhibitor, JNK-IN-8, in combination with dasatinib markedly improved the survival of mice with BCR-ABL induced B-ALL, as compared to the treatment with dasatinib alone. Our findings indicate that simultaneously targeting both BCR-ABL and JNK kinase might serve as a promising therapeutic strategy for Ph+ B-ALL.

Citation Format: Xinhua Xiao, Bo Jiao, Ruibao Ren. Combination therapy of BCR-ABL-positive B cell acute lymphoblastic leukemia by tyrosine kinase inhibitor dasatinib and c-Jun N-terminal kinase inhibition [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 4223.
Abstract 4239: Immunomodulatory effects of topotecan augment the effectiveness of chemotherapy-immunotherapy combination
Authors: Lee Jeong-Mi , Shin Kwang-Soo , Koh Choong-Hyun , Jeon Insu , Kang Tae-Seung , Song Boyeong , Kang Chang-Yuil
Published: September 2020

Combination therapy is a cornerstone of anticancer therapy, suggesting potential therapeutic benefit to nonresponders to single-agent treatment. BVAC, a B-cell-and-monocyte-based vaccine loaded with alpha-galactosylceramide(α-GC) and tumor antigen as reported in our previous studies, can be successfully combined with other treatments. Here we found out a novel synergistic regimen in TC-1 tumor model which combines BVAC with topotecan, a chemotherapeutic agent as a topoisomerase I inhibitor. We demonstrated some significant alteration in the immunoprofiles of topotecan-treated mice. We observed increased tumor recruitment of monocytes and CD8+ T cells, and decreased frequencies in regulatory T cells in tumor. Accordingly, we are currently investigating how topotecan-induced changes in immune cell composition could lead to improvement of antitumor effect. These data indicate that topotecan not only has cytotoxicity to tumor but can contribute to modulating tumor immune microenvironment. We anticipate that this study could provide a rationale for combination of cancer vaccine and chemotherapy and define future optimal treatment regimens in cancer patients.

Citation Format: Jeong-Mi Lee, Kwang-Soo Shin, Choong-Hyun Koh, Insu Jeon, Tae-Seung Kang, Boyeong Song, Chang-Yuil Kang. Immunomodulatory effects of topotecan augment the effectiveness of chemotherapy-immunotherapy combination [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 4239.
Abstract 4241: Regorafenib and nivolumab or pembrolizumab combination and ctDNA response assessment in refractory colorectal cancer
Authors: Fakih Marwan , Chevalier Dawnyel , Saluja Janelle , Lau Cecilia , Wang Chongkai
Published: September 2020

Introduction: Patients with microsatellite stable (MSS) metastatic colorectal cancer (MCRC) are considered resistant to immunotherapy with PD-1 and PD-L1 inhibitors. However, a recent study of regorafenib and nivolumab (REGONIVO) reported high response rates and a median progression free survival (PFS) of more than 6 months (mo) in a cohort of 24 Japanese patients (pts) with MSS chemo-resistant MCRC.

Methods: We evaluated the outcome of advanced MSS MCRC pts who were treated on a compassionate basis with PD-1 inhibitors in combination with regorafenib 80 mg PO QD x 21 days every 28 days in a single US center. We report their response rate (RR), PFS, and overall survival (OS).

Results: 16 pts (15 males) were treated with a combination of regorafenib + PD-1 inhibitors and were evaluable for response. 14 pts received regorafenib + nivolumab (5 with prior anti-PD-1 treatment) and 2 pts received regorafenib + pembrolizumab (both following pembrolizumab treatment). No treatment-related G3 or above toxicities were attributed to the combination therapy. No objective responses were noted on treatment. 11 pts had progressive disease (PD) and 5 pts experienced stable disease (SD) as best response at 2 mo. The duration of SD was 4 mo in 2 pts (1 pt with prior anti-PD1 exposure), 6 mo in 1 pt (prior anti-PD1 exposure), 5 mo+ in 2 pt (1 with prior anti-PD1 exposure). When limiting the analysis to the 9 regorafenib and checkpoint-inhibitor naïve patients, 2 pts were noted to have SD (4 months and 5 mo+). ctDNA assays were evaluated at baseline and at 4 weeks of treatment in 11 patients. All 9 patients with a rise in ctDNA at 4 weeks had PD at the 2-mo imaging point. 2 patients with declining ctDNA at 4 weeks experienced SD for 4 mo 5 mo+, each. OS analysis remains immature as the median follow-up is only 5 mo.

Conclusions: Regorafenib and nivolumab combination was associated with modest clinical activity in patients with MSS chemotherapy-resistant MCRC. This US-based small retrospective trial is at odds with the Japanese REGONIVO trial. ctDNA may represent a powerful tool for predicting progression prior to radiographic staging.

Citation Format: Marwan Fakih, Dawnyel Chevalier, Janelle Saluja, Cecilia Lau, Chongkai Wang. Regorafenib and nivolumab or pembrolizumab combination and ctDNA response assessment in refractory colorectal 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 4241.
Abstract 4254: Quality assessment platform for molecular pathology from non small cell lung cancer histopathology images using deep learning: A nationwide study of central laboratory in South Korea
Authors: Kim Tae-Jung , Kang Ho Chul , Lee Yonggeun , Seo Seokman , Kim Donghwan
Published: September 2020

Molecular testing became the most important part of lung cancer diagnostics. Integration of various molecular testing such as real-time PCR, fluorescent in situ hybridization (FISH) and next-generation sequencing into routine clinical practice and their systemic quality control is challenging to a pathologist and even more in the central laboratory. In this study, we trained a variant of DenseNet, a convolutional neural network model with the highest performance in image classification and suitable for quality assessment. Train dataset is composed of the arbitrarily classified dataset made and assessed by a professional pathologist during routine molecular test using whole-slide tissue images and molecular profiles including DNA density and the signal intensity of FISH obtained from 15,000 tests from lung cancer biopsies and resections, which were sent to the central laboratory for molecular studies from 45 hospitals nationwide. The performance of our platform is tested and compared to the quality assessment dataset made from each a general pathologist and a technician. The performance of our method for classifying DNA concentration and the signal intensity of FISH is comparable to that of a general pathologist and we are under improving its average area under the curve (AUC) over 0.95. This result suggests that deep learning model based on a convolutional neural network can assist pathologists for rigorous assessing quality of tissue morphology and signal of FISH for tissue samples with various molecular quality before or during the molecular test and, furthermore, can be applied to quality control of lung cancer molecular testing in a central laboratory.

Citation Format: Tae-Jung Kim, Ho Chul Kang, Yonggeun Lee, Seokman Seo, Donghwan Kim. Quality assessment platform for molecular pathology from non small cell lung cancer histopathology images using deep learning: A nationwide study of central laboratory in South Korea [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 4254.