bluuebunny/crossref_metadata_2025 huggingface dataset:
Isoflurane Regulates Proliferation, Apoptosis, and Inflammatory Response of Lipopolysaccharide-Induced Human Astrocyte through the miR-206/BDNF Axis
Authors: Wang Jianying , Liu Zhiyuan , Wang Xue , Liu Yu
Published: March 2020

Objective. To investigate the effect of isoflurane (ISO) on the proliferation, apoptosis, and inflammatory response of lipopolysaccharide- (LPS-) induced normal human astrocytes (NHAs) by regulating the miR-206/BDNF axis. Methods. NHA proliferation activity was measured by MTT; NHA apoptotic rates were measured by Annexin V-FITC/PI; western blotting was used to measure the BDNF expression; ELISA was used to measure the IL-6, IL-1β, and TNF-α expression in NHAs; qPCR was used to measure the expressions of miRNAs that are related to NHAs proliferation and apoptosis; dual-luciferase reporter was constructed to validate the targeting relationship between miR-206 and BDNF. Results. LPS increased the proliferation activity and decreased the apoptosis rate of NHAs which were effectively reversed by the ISO (p<0.05); LPS significantly inhibited the expression of miRNAs related to proliferation and apoptosis in NHAs (p<0.05, p<0.01), whereas ISO significantly increased the expression of miR-206 (p<0.01) by downregulating the expression of BDNF, thus inhibiting NHA proliferation and inflammatory response and enhancing apoptosis. Conclusion. ISO can inhibit the expression of BDNF by upregulating the expression of miR-206, thereby inhibiting the proliferation and inflammatory response of NHAs and promoting its apoptosis.
Reduction of an Unusual Salter-Harris Type IV Fracture of the Ulna
Authors: Marrannes Stephanie , Lambrecht Delphine , Decramer Arne
Published: March 2020

We report the case of a 14-year-old boy with an isolated Salter-Harris type IV physeal fracture of the distal ulna. Following failed closed reduction, transition to open reduction and pin fixation was required. Six-month follow-up showed a favourable clinical evolution. Evidence suggests that long-term follow-up is needed because of the increased risk of premature physeal closure and subsequent growth disturbances associated with this type of injury.
Omega, Sadhana, and PI Polynomials of Quasi-Hexagonal Benzenoid Chain
Authors: Idrees Nazeran , Saif Muhammad Jawwad , Nasir Sumiya , Farooq Fozia Bashir , Rauf Asia , Ashfaq Fareeha
Published: March 2020

Counting polynomials are important graph invariants whose coefficients and exponents are related to different properties of chemical graphs. Three closely related polynomials, i.e., Omega, Sadhana, and PI polynomials, dependent upon the equidistant edges and nonequidistant edges of graphs, are studied for quasi-hexagonal benzenoid chains. Analytical closed expressions for these polynomials are derived. Moreover, relation between Padmakar–Ivan (PI) index of quasi-hexagonal chain and that of corresponding linear chain is also established.
Acquired Resistance to Alectinib in ALK-Rearranged Lung Cancer due to ABCC11/MRP8 Overexpression in a Clinically Paired Resistance Model
Authors: Funazo Tomoko , Tsuji Takahiro , Ozasa Hiroaki , Furugaki Koh , Yoshimura Yasushi , Oguri Tetsuya , Ajimizu Hitomi , Yasuda Yuto , Nomizo Takashi , Sakamori Yuichi , Yoshida Hironori , Kim Young Hak , Hirai Toyohiro
Published: March 2020

Alectinib is used as a first-line treatment for anaplastic lymphoma kinase (ALK)-rearranged non–small cell lung cancer (NSCLC). Whereas other ALK inhibitors have been reported to be involved in resistance to ATP-binding cassette (ABC) transporters, no data are available regarding the association between resistance to alectinib and ABC-transporters. To investigate whether ABC-transporters contribute to alectinib resistance, ABC-transporter expression in alectinib-resistant cell lines derived from a patient with ALK-rearranged NSCLC and from H2228 lung cancer cells was evaluated and compared with that in each parent cell type. ATP-binding cassette subfamily C member 11 (ABCC11) expression was significantly increased in alectinib-resistant cell lines compared with that in alectinib-sensitive lines. ABCC11 inhibition increased sensitivity to alectinib in vitro. ABCC11-overexpressing cells were established by transfection of an ABCC11 expression vector into H2228 cells, while control cells were established by transfecting H2228 cells with an empty vector. ABCC11-overexpressing cells exhibited decreased sensitivity to alectinib compared with that of control cells in vitro. Moreover, the tumor growth rate following alectinib treatment was higher in ABCC11-overexpressing cells than that in control cells in vivo. In addition, the intracellular alectinib concentration following exposure to 100 nmol/L alectinib was significantly lower in the ABCC11-overexpressing cell line compared with that in control cells. This is the first preclinical evidence showing that ABCC11 expression may be involved in acquired resistance to alectinib.
Abstract A001: Colorectal cancer cell-derived exosomes containing miR-183-5p regulate angiogenesis via FOXO1
Authors: SHANG ANQUAN , Gu Chenzheng , Wu Junlu , Ji Ping , Sun Zujun , Li Dong
Published: March 2020

Objective: Exosomes acted as important role in tumor proliferation and tumor microenvironment modulation, including colorectal cancer (CRC). However, the inhibitory effect of CRC cells -derived exosomes in angiogenesis in vascular endothelial cell has not been fully discussed. In this study, the microRNA-183-5p (miR-183-5p) that promote the development of CRC, as well as the exosome secreted from the CRC cells were investigated. Methods: Microarray analysis was employed to determine the differentially expressed miRNAs. Exosomes derived from CRC cells were isolated and identified. Meanwhile, exosomes derived from CRC cells were isolated and then co-cultured with HMEC-1 cells to identify the role of exosomes in angiogenesis. Further, the effects of CRC cell-derived exosomal miR-183-5p on proliferation, invasion and tube formation abilities of HMEC-1 cells were assessed. The preventative effect of exosomal miR-183-5p in vivo was measured in nude mice. Results: It was found that FOXO1 was downregulated while miR-183-5p was upregulated in CRC. Additionally, the inhibition of miR-183-5p was suggested to suppress proliferation, invasion and tube formation abilities of HMEC-1 cells through upregulating FOXO1. Then, in vitro experiments demonstrated that CRC cell-derived exosomes overexpressing miR-183-5p contributed to an enhanced proliferation, invasion and tube formation abilities of HMEC-1 cells. Furthermore, in vivo experiments confirmed the promotive effects of CRC cell-derived exosomes overexpressing miR-183-5p. Conclusion: Our study demonstrates that the CRC cell-derived exosomes overexpressing miR-183-5p aggravates CRC through the regulation of FOXO1. Additionally, exosomes overexpressing miR-183-5p might be a potential treatment biomarker for CRC.

Citation Format: ANQUAN SHANG, Chenzheng Gu, Junlu Wu, Ping Ji, Zujun Sun, Dong Li. Colorectal cancer cell-derived exosomes containing miR-183-5p regulate angiogenesis via FOXO1 [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A001. doi:10.1158/1535-7163.TARG-19-A001
Abstract A015: A kinetic analysis of tumor gene expression and cellular changes following immune checkpoint inhibition in the MC38 colon carcinoma model
Authors: Fadden Patrick , Bui Thi , Davis Kelli , Hurtado David , Belle Ian , Krontz Katherine , Hauser Jacob , Sokolowski Cristina , Li Na , Takayama Kyle , Marval Paula Miliani de , Harris David , Wood Edgar
Published: March 2020

The clinical success of immune checkpoint therapies such as PD-1, PD-L1, and CTLA-4 inhibitory antibodies has stimulated interest in a wide range of approaches to cancer immunotherapy. Development of these new modalities requires robust, well characterized preclinical animal models to evaluate efficacy and potential toxicities. Preclinical efficacy assessments of novel immune-oncology therapies requires a functional immune system which limits the usefulness of traditional xenograft models and drive the use of humanized (human immune cell engrafted animals) and syngeneic model systems. Syngeneic mouse tumors are popular model systems in which to evaluate these novel therapies due the presence of fully intact mouse immune system and ease of accessibility. The MC38 mouse colon cancer model is popular for efficacy assessment studies due to its responsiveness to typical immune checkpoint inhibitors (ICI). We endeavored to characterize the kinetic immune response to checkpoint inhibitors (anti-PD-1 + anti-CTLA-4) in MC38 tumors using flow cytometry and gene expression analysis. Gene expression analysis provided a signature of gene changes that correlate to immune driven changes in tumor growth and may be used in preclinical pharmacodynamics studies as evidence of mode of action for other novel immune-oncology therapies. In addition, gene over-representation analysis highlighted the presence and involvement of B cell populations in the MC38 tumor environment which was supported by IHC data that show distinctive B cell staining in what resemble tumor associated tertiary lymphoid structures. These findings provide a greater understanding of the ICI response in the MC38 tumor model and may inform mode of action study designs for other novel immuno-oncology therapies.

Citation Format: Patrick Fadden, Thi Bui, Kelli Davis, David Hurtado, Ian Belle, Katherine Krontz, Jacob Hauser, Cristina Sokolowski, Na Li, Kyle Takayama, Paula Miliani de Marval, David Harris, Edgar Wood. A kinetic analysis of tumor gene expression and cellular changes following immune checkpoint inhibition in the MC38 colon carcinoma model [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A015. doi:10.1158/1535-7163.TARG-19-A015
Abstract A021: Genipin enhanced apoptosis of gastric cancer by mitochondrial dysfunction and downregulation of Stat3/JAK2/Mcl-1 signaling pathway
Authors: Jo Min Jee , Jeong Soyeon , Yun Hye Kyeong , Kim Dae Yeong , Kim Bo Ram , Kim Jung Lim , Na Yoo Jin , Park Seong Hye , Jeong Yoon A , Kim Bu Gyeom , Ashktorab Hassan , Smoot Duane T , Heo Jun Young , Han Jeongsu , Lee Dae-Hee , Oh Sang Cheul
Published: March 2020

Genipin is known as a compound derived from gardenia fruit extract. Although Genipin is known to have an anti-tumor effects in various cancers, its effect and mechanism are unknown in gastric cancer. In our studies, Genipin induced apoptosis in gastric cancer cells including AGS and MKN45 cells. Genipin reduced Mcl-1 mRNA levels and confirmed with transfection of Mcl-1 siRNA. Furthermore, We found that phospho Stat3 is regulated by Genipin through proteome profiler phosphor-kinase array. In addition, protein level of phosphor JAK2 decreased in Genipin treatment. The results elucidate that Stat3/JAK2/Mcl-1 signaling pathway was suppressed with Genipin treatment in gastric cancer cells. Mcl-1 is known to be closely related to the function of mitochondria. Genipin has been shown to result in the collapse of mitochondrial function such as mitochondrial membrane potential. In conclusion, Genipin induced apoptosis by suppressing Stat3/JAK2/Mcl-1 pathway and led to mitochondrial dysfunction. Our finding implies that a novel mechanism of anti-cancer effect of Genipin in gastric cancer can be identified.

Citation Format: Min Jee Jo, Soyeon Jeong, Hye Kyeong Yun, Dae Yeong Kim, Bo Ram Kim, Jung Lim Kim, Yoo Jin Na, Seong Hye Park, Yoon A Jeong, Bu Gyeom Kim, Hassan Ashktorab, Duane T Smoot, Jun Young Heo, Jeongsu Han, Dae-Hee Lee, Sang Cheul Oh. Genipin enhanced apoptosis of gastric cancer by mitochondrial dysfunction and downregulation of Stat3/JAK2/Mcl-1 signaling pathway [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A021. doi:10.1158/1535-7163.TARG-19-A021
Abstract A022: Gasdermin E suppresses tumor growth by activating antitumor immunity
Authors: Zhang Zhibin , Zhang Ying , Xia Shiyu , Kong Qing , Li Shunying , Liu Xing , Junqueira Caroline , Ansara James , Sengupta Satyaki , Yao Yandan , Wu Hao , Lieberman Judy
Published: March 2020

Caspase-3 cleaves gasdermin E (GSDME) and converts noninflammatory apoptosis to inflammatory cell death (pyroptosis) in GSDME-expressing cells. GSDME expression is suppressed in many cancer lines and mutated in some others, and reduced GSDME is associated with metastasis and decreased survival in breast cancer, suggesting GSDME might be a tumor suppressor. Here we show reduced GSDME function of 7 of 8 tested cancer-associated mutations. Gsdme knockout in GSDME-expressing tumors enhances, while ectopic expression in Gsdme-repressed tumors inhibits, tumor growth. The GSDME tumor-suppressive effect is cell-extrinsic, mediated by cytotoxic lymphocyte killing and abrogated in perforin-deficient mice or mice lacking lymphocytes. GSDME expression enhances tumor-associated macrophage phagocytosis and the number and functions of tumor-infiltrating NK and CD8+ T lymphocytes. Killer cells also activate GSDME-mediated pyroptosis independently of caspase activation when Granzyme B directly cleaves GSDME at the same site as caspase-3 (Asp270). Thus, GSDME suppresses tumor growth by activating anti-tumor immunity.

Citation Format: Zhibin Zhang, Ying Zhang, Shiyu Xia, Qing Kong, Shunying Li, Xing Liu, Caroline Junqueira, James Ansara, Satyaki Sengupta, Yandan Yao, Hao Wu, Judy Lieberman. Gasdermin E suppresses tumor growth by activating antitumor immunity [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A022. doi:10.1158/1535-7163.TARG-19-A022
Abstract A055: Knockdown of E-cadherin gene in cervical cancer cells: An in vitro model for cancer stem cell phenotype?
Authors: Bhattacharyya Shalmoli , Sharma Anuka , Pal Arnab , Srinivasan Radhika
Published: March 2020

Background: Cervical cancer is one of the leading causes of cancer related morbidity and mortality amongst women, especially in developing countries. Recent studies suggest that Cancer stem cells (CSC) are critically involved in regulating the chemo resistant behavior of cancer cells and a relapse of the disease. So targeting CSC-like cells along with the bulk tumor cells can be a promising approach for therapeutics. In this context, we demonstrated that knockdown of E-cadherin gene in cervical cancer cells can generate stem cell-like phenotype and induce epithelial to mesenchymal transition. Interestingly, knockdown of E-cadherin gene in normal keratinocytes did not induce CSC-like cells. Methods: In our study the CSC phenotype cells were isolated by side population (SP) assay from cervical cancer cells and immortalized keratinocytes. The expression of stem cell marker and EMT genes were confirmed by qRT-PCR and immunostainings. Further, we performed SiRNA-mediated knockdown of E-Cadherin (E-Cad) and studied the tumorsphere formation, metastasis and invasion ability of cervical cancer cells. Next, using RTPCR based gene expression analysis and tumorsphere formation we studied the stem–like properties of cancer cells. We also performed dose dependent cell viability assay using clinically relevant drugs and inhibitors to analyze the drug resistance behavior of the tumorspheres (cervical organoids). Results: We found that E-Cad knockdown induces EMT in cervical cancer cells with stem cell-like characteristics along with enhanced tumorsphere formation, metastasis and invasion ability. These cells not only possess CSC-like phenotype at transcriptional and translational level but also functionally as shown by drug resistance behavior studies. Conclusion: Thus, we believe that this can be a promising model to study CSC-like characteristics in-vitro in cervical cancer to target the molecular pathways for therapeutics.

Citation Format: Shalmoli Bhattacharyya, Anuka Sharma, Arnab Pal, Radhika Srinivasan. Knockdown of E-cadherin gene in cervical cancer cells: An in vitro model for cancer stem cell phenotype? [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A055. doi:10.1158/1535-7163.TARG-19-A055
Abstract A057: The MEK1/2 pathway as a therapeutic target in high-grade serous ovarian carcinoma
Authors: Chesnokov Mikhail , Yousif Abdelrahman , Chefetz ilana
Published: March 2020

Rationale: High-grade serous ovarian carcinoma (HGSOC) is the deadliest of gynecological cancers due to high rates of recurrence and acquired chemoresistance. Recurrent tumors are suggested to originate from cancer stem-like cells (CSCs) which have been shown to have high MEK1/2 activity signaling pathway to survive and proliferate. Present work investigates the potential of selective MEK1/2 inhibition as a CSC-targeting HGSOC therapy approach. Methods: MEK1/2 pathway activity was evaluated in clinical HGSOC tissue samples and ovarian cancer cell lines using tissue microarray-based immunohistochemistry, immunoblotting and RT-qPCR. OVCAR8 and PEO4 HGSOC cell lines were used to assess the effect of MEK1/2 inhibition on cell viability, proliferation rate and stem-like characteristics in vitro. Mouse xenografts were used to investigate MEK1/2 inhibition effect on tumor growth in vivo. Drug washout experimental model was used to study the lasting effects of MEK1/2 inhibition therapy. Results: MEK1/2 signaling is active in majority of HGSOC tissue samples and cell lines and is further stimulated by cisplatin treatment. MEK1/2 inhibitor trametinib drastically inhibits activity of MEK1/2 downstream signaling, causes prominent cell cycle arrest in G1/0-phase in vitro and reduces tumor growth rate in vivo, but does not induce cell death. Cells treated with trametinib display high CD133+ fraction and increased expression of stemness-associated genes. Transient trametinib treatment causes long-term increase in high ALDH1 activity cells that possess capability of surviving and growing in non-adherent conditions. Conclusions: MEK1/2 inhibition in HGSOC cells efficiently inhibits cell proliferation and tumor growth and therefore may be promising approach to suppress ovarian cancer cell growth. As MEK1/2 inhibition promotes stem-like properties, including ALDH activity, it should be used in combination with CSC-targeting drugs, especially ALDH1 inhibitors.

Citation Format: Mikhail Chesnokov, Abdelrahman Yousif, ilana Chefetz. The MEK1/2 pathway as a therapeutic target in high-grade serous ovarian carcinoma [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A057. doi:10.1158/1535-7163.TARG-19-A057
Abstract A060: Inhibition of Aurora B kinase activity triggers senescence that can be bypassed by blocking p53 and RB function, promoting replication stress
Authors: Andrews Ariel , Kumar Ramyashree Prasanna , Nazareth Deborah , Ehmann Anna , Hooper John , McMillan Nigel , Gabrielli Brian
Published: March 2020

Aurora B kinase has a major role in regulating progression through mitosis and partitioning the replicated genome at exit from mitosis and cytokinesis. We have previously reported that Aurora A and B inhibitors selectively targeted HPV-driven tumours, and that the HPV E7 oncogene is a major determinant of this selectivity. Another group has shown that sensitivity to Aurora B inhibitor AZD2811 is dependent on loss of the retinoblastoma protein (RB). Here we have investigated the outcomes of inhibition of Aurora B using selective and pan Aurora inhibitors. We show that inhibition of Aurora A and B promotes more cell death, although a subset of cells are protected from the cytotoxic effects as a consequence of Aurora A inhibition slowing the cell cycle. Aurora B inhibition promotes senescence, although this requires at least two cycles of failed cytokinesis in all cell types, including primary fibroblasts. Loss of RB function either by HPV E7 or CDK4 R24C mutant over-expression, and loss of p53 function have somewhat different effects on the outcomes of Aurora B inhibition, although both reduced the proportion of cells that entered senescence. Loss of either also promoted replication stress which was not observed in RB and p53 proficient cells, the level of replication stress was not enhanced by Aurora A co-inhibition. Selective Aurora B inhibitor was also less toxic to proliferating PBMC derive CD3+ T cells than the pan Aurora inhibitor. Together these data indicate that short term treatment with Aurora B selective inhibitor is sufficient to promote cell cycle arrest and senescence in RB and p53 proficient cells, although proliferating T cells appear to tolerate this. RB and p53 pathway defective cells are less sensitive to this senescence trigger, but undergo replication stress.

Citation Format: Ariel Andrews, Ramyashree Prasanna Kumar, Deborah Nazareth, Anna Ehmann, John Hooper, Nigel McMillan, Brian Gabrielli. Inhibition of Aurora B kinase activity triggers senescence that can be bypassed by blocking p53 and RB function, promoting replication stress [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A060. doi:10.1158/1535-7163.TARG-19-A060
Abstract A065: Differential sensitivity of normal and tumor organoids to targeted therapies
Authors: Madej Mariusz , Zuvirie Cinthya Del Angel , Mateos Jara García , Price Leo , Daszkiewicz Lidia , Yan Kuan , Herpers Bram
Published: March 2020

Background: Advances in the culture of cells from human tissues have enabled the generation of normal and tumor organoid pairs derived from the same patient. These organoid cultures increase the scope for predicting responses to novel cancer therapeutics in patients. Here we present a high throughput 3D human intestinal organoid culture platform combined with high content image-based analysis. This platform allows visualization and quantification of responses beyond conventional cell viability measurements, including those associated with tumor killing, cell cycle arrest, toxicity and epithelial permeability and discriminates therapeutic and adverse responses in healthy tissue and tumor. Methods: Intestinal organoids, obtained from HUB Organoid Technology, cultured in natural extracellular matrix scaffolds show gene expression patterns, differentiation and functional characteristics that closely resemble the in vivo biology. To investigate the effect of standard-of-care (SoC) treatments, novel targeting antibodies and small molecules on tumor growth suppression and killing as well as function, formation and integrity of the gut epithelium, we tested a panel of normal and colorectal cancer organoids with a broad heterogeneity of mutations. 3D imaging and analysis was performed and responses in normal vs. tumor were compared. Results: High content 3D image analysis of the organoid enabled sensitive detection of treatment-induced and compound-specific morphological changes such as (inhibition of) growth, development, lumen formation, epithelial integrity and cell death. This enabled distinction between mechanisms of action and distinct effects and sensitivities in normal epithelium and tumor tissue from the same patient. Conclusion: 3D image analysis of in vitro cultured organoids represents a rapid and biologically relevant approach to test various anti-cancer-therapeutic modalities, including antibodies, antibody-drug conjugates and small molecules. Evaluating effects on healthy and tumor tissue from the same patient can give insight into potential toxicities and therapeutic window.

Citation Format: Mariusz Madej, Cinthya Del Angel Zuvirie, Jara García Mateos, Leo Price, Lidia Daszkiewicz, Kuan Yan, Bram Herpers. Differential sensitivity of normal and tumor organoids to targeted therapies [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A065. doi:10.1158/1535-7163.TARG-19-A065
Abstract A066: Role of the NRF2 signaling pathway in sustaining chemoresistance in medulloblastoma
Authors: Rruga Fatlum , Mariotto Elena , Persano Luca , Rampazzo Elena , Luchini Alessandra , Liotta Lance A , Viola Giampietro , Bortolozzi Roberta
Published: March 2020

Nrf2, a key orchestrator of cell defense mechanisms against oxidative stress and xenobiotic insults, has been linked to chemotherapy resistance and recurrence in many human cancers. Through the establishment of an in vitro model of chemoresistance, we investigated the role of Nrf2 in sustaining resistance to chemotherapy in medulloblastoma (MB), a highly aggressive pediatric brain tumor intrinsically characterized by fast growth, high invasiveness and resistance to treatments. MB DAOY cells were weekly exposed to a cocktail of drugs commonly used in MB treatment, vincristine, etoposide, cisplatin and cyclophosphamide (VECC) inducing the emergence of a resistant phenotype. After 5 cycles of VECC treatment we obtained a culture of cells resistant to subsequent chemotherapy treatments. We observed increased Nrf2 levels and activity in therapy-exposed cells already from the first received dose of VECC. Nrf2 levels remained high throughout all the rest of chemotherapy cycles and interestingly, its transcriptional activity spiked up when resistance to chemotherapy emerged, as demonstrated by the increase of antioxidant response element (ARE)-driven luciferase activity and the expression of target genes. The knock down of Nrf2 in resistant cells, by the use of specific siRNA, was able to restore chemosensitivity, suggesting a role of Nrf2 signaling pathway in the onset of chemotherapy resistance in MB cells. Resistant cells showed a higher ROS (reactive oxygen species) scavenging capacity after stimulation with hydrogen peroxide. Nrf2 silencing partially reverts the ability of resistant cells to manage oxidative stress, supporting the hypothesis of Nrf2 involvement in the ROS scavenging potential of VECC resistant MB cells. In addition to directly regulate ROS-scavenging enzymes, Nrf2 has been described to control cell metabolism that maintain redox homeostasis in cancer stem cells. Among the Nrf2 target genes upregulated in DAOY resistant cells we found the upregulation of PFKFB-3, G6PD and TKT. The increased expression of this enzymes suggests a metabolic reprogramming in resistant cells. Our data support the involvement of Nrf2 pathway in MB response to VECC treatment and show that the upregulation of this detoxifying system sustains MB resistance to chemotherapy.

Citation Format: Fatlum Rruga, Elena Mariotto, Luca Persano, Elena Rampazzo, Alessandra Luchini, Lance A Liotta, Giampietro Viola, Roberta Bortolozzi. Role of the NRF2 signaling pathway in sustaining chemoresistance in medulloblastoma [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A066. doi:10.1158/1535-7163.TARG-19-A066
Abstract A100: Tumor-targeted and pH-shiftable nanoparticles to encapsulate oxaliplatin and microRNA for colorectal cancer therapy
Authors: Lo Yu-Li , Chen Y.C. , Chang C.H. , Tseng W.H. , Wang C.S.
Published: March 2020

Colorectal cancer is a popular cancer type all over the world. Among various treatment regimen, oxaliplatin (Oxa) is approved in combination with 5-fluorouracil (5-FU) and leucovorin (FOLFOX) as the first-line therapy for colorectal cancer of metastatic or advanced stage. MicroRNAs (miRs) may target genes critical for regulating tumor proliferation and invasion. Among different miRs, miR-c has been proved to induce apoptosis, inhibit proliferation and epithelial-mesenchymal transition of cancer cells. Peptide-modified nanoparticles, which are capable of increasing cellular uptake and targeting to specific receptors, are well suitable for delivering Oxa and miR. Moreover, coating pH-sensitive polymers to the outer layer of peptide-modified nanoparticles may enhance specific targeting to the acidic tumor sites. Physicochemical characterization, cellular uptake, cytotoxicity, migration assay, and western blotting studies of these nanoparticle formulations have been performed. Our results indicated that these pH-sensitive and peptide-modified nanoparticles displayed a narrow size distribution. The cellular uptake of peptide-modified nanoparticles was found to be greater than that of naked nanoparticles. Besides, the cellular uptake of pH-sensitive nanoparticles was significantly increased at pH 6.0 compared to that at pH 7.4, suggesting the successful pH-responsive property of these delivery systems. Furthermore, the combined treatment of miR-c and Oxa by peptide-modified nanoparticles increased the cytotoxicity of Oxa to colorectal cancer cells. We demonstrated that the upregulation of apoptosis pathway and the downregulation of β-catenin pathway were accounted for the efficient antitumor effect. In conclusion, the delivery of miR-c and Oxa in pH-sensitive and peptide-modified nanoparticles may provide a novel and potential strategy for colorectal cancer treatment.

Citation Format: Yu-Li Lo, Y.C. Chen, C.H. Chang, W.H. Tseng, C.S. Wang. Tumor-targeted and pH-shiftable nanoparticles to encapsulate oxaliplatin and microRNA for colorectal cancer therapy [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A100. doi:10.1158/1535-7163.TARG-19-A100
Abstract A102: pHLIP technology for targeting acidity at surface of tumor cells and extracellular and intracellular delivery of imaging and therapeutic agents
Authors: Reshetnyak Yana K , Andreev Oleg A , Engelman Donald
Published: March 2020

pH-Low Insertion Peptides (pHLIP® peptides) are a platform technology for targeting of acidity at the surface of individual cells within tumors. Most proteins that are located across membranes have one or more helical segments that cross the lipid bilayer, and studies of these membrane proteins revealed pHLIPs, which could fold from disorganized (unstructured) states outside of a membrane to form a helix across the membrane in response to local acidity. Because tumor cell surfaces are acidic as a result of their active metabolism, versions of pHLIP® peptides were extensively investigated and tested as possible tumor targeting agents. Our studies showed that an extensive variety of cargoes can be targeted to cell surfaces and that a surprising range of cargoes can be delivered across membranes into cells if their surfaces are acidic, creating opportunities to detect and treat tumors. pHLIP® peptides can be used to target and tether cargo molecules to the surfaces of cells in low pH environments of acidic diseased tissues. The cargo can be an optical marker, a PET, a SPECT, or a MR imaging agent, or an antigen or a protein delivered to induce certain cellular or immune responses. We are translating pHLIP®-based imaging agents to the clinic for diagnostic PET imaging, fluorescence-guided surgical procedures and photo-acoustic pre-operative imaging. pHLIP® peptides can also be used for the intracellular delivery of payloads, facilitating the translocation of therapeutic cargoes across the membranes of cells with low extracellular pH, such as those cells found in acidic diseased tissue. These payloads are conjugated to the membrane-inserting end of pHLIP® peptide, typically via a link that is unstable inside the cell. Thus, pHLIP® peptide targets a payload to acidic diseased cells and flips the cargo across the plasma membrane, releasing it in the cytoplasm. Therapeutic cargoes may include toxins, metabolic activators/inhibitors, chemotherapeutic agents, or agents to alter gene expression. *YKR, OAA, and DME are founders of pHLIP, Inc. and have financial interests in the company.

Citation Format: Yana K Reshetnyak, Oleg A Andreev, Donald Engelman. pHLIP technology for targeting acidity at surface of tumor cells and extracellular and intracellular delivery of imaging and therapeutic agents [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A102. doi:10.1158/1535-7163.TARG-19-A102
Abstract A107: Small Molecule Degraders of the Estrogen Receptor (SERDs): Optimization of the tricyclic indole scaffold beyond AZD9496
Authors: Barlaam Bernard , Breed Jason , Carbajo Rodrigo J , Gangl Eric , Hughes Samantha , Morrow Christopher J , Moss Thomas A , Polanski Radoslaw , Nissink Willem M , O'Donovan Daniel , Varnes Jeffrey , Yang Bin , Scott James S
Published: March 2020

The estrogen receptor alpha (ERα) is expressed in >70% of breast cancers and is a clinically validated target in oncology.1 Anti-hormonal therapies that directly block ER function (e.g., tamoxifen) or therapies that block the production of estrogen itself (e.g., aromatase inhibitors) have proven to be effective treatments of the disease. Further advances have been made with the development of SERDs (Selective Estrogen Receptor Degraders) such as fulvestrant which both antagonise ERα-driven tumor cell growth and cause degradation of the ERα receptor.2 We previously disclosed the identification of a tricyclic indole scaffold that led to the orally active clinical candidate AZD9496.3 Additional work to identify novel chemotypes including phenol 14 and indazole 25 was also disclosed together. The work previously disclosed relied on the presence of the acrylic acid for efficient degradation of the estrogen receptor. In this poster we will discuss the replacement of the acrylic acid on the tricyclic indole scaffold with amines (e.g. compound 3). Compound 3 is a degrader of the estrogen receptor in the MCF-7 cell line (pIC50 8.4). Further optimisation of lead structure 3 led to more potent selective degraders of the estrogen receptor in multiple cell lines (e.g. pIC50 >10 in MCF-7) with suitable properties for oral absorption (e.g. oral AUC 44 μM.h in mice at 20 mg/kg). We will discuss some of the medicinal chemistry challenges that were faced along the way and the optimisation strategy (use of NMR derived solution phase conformations, understanding of the binding mode in the estrogen receptor by X-ray crystallography). ol>

Citation Format: Bernard Barlaam, Jason Breed, Rodrigo J Carbajo, Eric Gangl, Samantha Hughes, Christopher J Morrow, Thomas A Moss, Radoslaw Polanski, Willem M Nissink, Daniel O'Donovan, Jeffrey Varnes, Bin Yang, James S Scott. Small Molecule Degraders of the Estrogen Receptor (SERDs): Optimization of the tricyclic indole scaffold beyond AZD9496 [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A107. doi:10.1158/1535-7163.TARG-19-A107
Abstract A111: AF8c, a pan-ErbB family inhibitor induces apoptotic cell death by stimulating DR5/Nrf2 via ROS in colorectal cancer cells
Authors: Jeong Soyeon , Farag Ahmed Karam , Yun Hye Kyeong , Jeong Yoon A , Kim Dae Yeong , Jo Min Jee , Park Seong Hye , Kim Bo Ram , Kim Jung Lim , Lee Dae-Hee , Roh Eun Joo , Oh Sang Cheul
Published: March 2020

Multifunctional, antiproliferative small molecules are deemed to offer pharmacodynamic and pharmacokinetic benefits over combination therapy, in addition to reducing toxicity, cancer resistance, and therapy costs. In this study, we conducted an in vitro cellular screen of our recently reported series of EGFR/HER2 inhibitors. Due to its preferentially apoptosis triggering in colorectal cancer (CRC) cells compared to normal colon cells, the compound AF8c was selected for further evaluation. Analysis of the AF8c mode of action in CRC cells revealed that it mediates apoptosis via the generation of endoplasmic reticulum (ER) stress and reactive oxygen species (ROS), as well as selective activation of nuclear respiratory factor 2 alpha subunit (Nrf2) and death receptor 5 (DR5), but not DR4. Silencing of DR5 attenuated the expression levels of Nrf2 and partially inhibited AF8c-induced apoptosis. Additionally, upregulation of Nrf2 by AF8c evoked apoptosis through a decrease in antioxidant levels. Treatment of mice with AF8c also resulted in the upregulation of DR5, Nrf2, and CHOP proteins, subsequently leading to a significant decrease in tumor burden. Overall, our results suggest that AF8c-induced apoptosis may be associated with DR5/Nrf2 activation through ER stress and ROS generation in CRC. These findings indicate that AF8c represents a promising polypharmacological molecule for the treatment of human CRC. Moreover, it could also be a potential starting point for the understanding of the structural features of quinazoline-based ErbB family inhibitors targeting the TRAIL cascade.

Citation Format: Soyeon Jeong, Ahmed Karam Farag, Hye Kyeong Yun, Yoon A Jeong, Dae Yeong Kim, Min Jee Jo, Seong Hye Park, Bo Ram Kim, Jung Lim Kim, Dae-Hee Lee, Eun Joo Roh, Sang Cheul Oh. AF8c, a pan-ErbB family inhibitor induces apoptotic cell death by stimulating DR5/Nrf2 via ROS in colorectal cancer cells [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A111. doi:10.1158/1535-7163.TARG-19-A111
Abstract B007: OKI-179 is a novel, oral, class I specific histone deacetylase inhibitor in phase 1 clinical trials
Authors: Diamond Jennifer R , Kagihara Jodi A , Liu Xuedong , Gordon Gilad , Heim Amy M , Winkler James , DeMattei John A , Piscopio Anthony D , Eckhardt S. Gail
Published: March 2020

Multiple histone deacetylase (HDAC) inhibitors are approved for use as single agents in hematological cancers, including T-cell lymphomas and multiple myeloma. However, these compounds have failed to address unmet needs in other hematological malignancies and solid tumors, due in part to the promiscuous inhibitory properties of pan inhibitors, a lack of suitable therapeutic indices for combination therapy, and, in some cases, a non-oral route of administration. Of the approved agents, romidepsin is the most potent and selective, but is limited by its need for IV infusion and therapeutic index. With these challenges in mind, our team designed OKI-179, a novel, oral pro-drug analog of largazole, a compound reminiscent of the romidepsin-depsipeptide class of natural products. OKI-179 demonstrates potent inhibition of the Class 1 HDACs, HDAC 1,2,3 (IC50 = 1.2, 2.4, 2.0 nM, respectively), with no significant inhibition of Class IIa HDACs. It shows potent antiproliferative and apoptotic effects against a wide range of cell lines, activity in xenograft cancer models, and good oral PK properties in mouse, rat and dog models. OKI-179 inhibits the growth in mice of colorectal (HCT-116) and triple negative breast (MDA-MB-231) xenograft models potently and dose-dependently as a single agent. OKI-179 potently and dose-dependently causes the upregulation of MHC Class 1 in B16F10 murine melanoma cells; patients with low MHC presentation respond poorly to checkpoint inhibitors. Importantly, the combination of OKI-179 and a murine anti-PD1 agent demonstrates significant synergistic tumor growth inhibition in the syngeneic CT26 murine CRC model and in a genetic model of DLBCL. OKI-179 is currently being investigated in a first-in-human phase 1 dose-escalation trial, with the goals of determining safety, MTD, RP2D, PK, PD, and preliminary efficacy signals. In the first dosing cohorts, the drug was well-tolerated, achieved good exposure following oral administration, and preliminary signs of on-target PD effects were observed. An update of the clinical experience will be presented and discussed, along with translational data that will guide subsequent clinical development.

Citation Format: Jennifer R Diamond, Jodi A Kagihara, Xuedong Liu, Gilad Gordon, Amy M Heim, James Winkler, John A DeMattei, Anthony D Piscopio, S. Gail Eckhardt. OKI-179 is a novel, oral, class I specific histone deacetylase inhibitor in phase 1 clinical trials [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B007. doi:10.1158/1535-7163.TARG-19-B007
Abstract B031: OneThree Biotech’s artificial intelligence platform for optimizing drug development
Authors: Gilvary Coryandar , Madhukar Neel , Elemento Olivier
Published: March 2020

Recently, it has been shown how artificial intelligence (AI) has the possibility to dramatically shorten the drug development pipeline by identifying insights that may have otherwise been missed. However, one major critique of these methods has been their black-box nature and the lack of mechanistic biology. To address these issues, OneThree Biotech has developed an extensive platform of biology-driven AI approaches that accelerate early stage drug development. Beginning with novel target identification, we present ECLIPSE, an AI approach that combines genomic, cell line and experimental design features to identify essential genes within cancer cell lines, based upon CRISPR and shRNA loss-of-function screenings. We demonstrated that ECLIPSE could accurately identify known and potential cancer targets, as well as be used to determine drug efficacy in the clinic. In cases where a compound’s target is unknown, we introduce BANDIT, a Bayesian model that combines divergent data sources to predict the targets and mechanisms for small molecules with unprecedented accuracy and versatility. Using BANDIT, we successfully identified a novel class of microtubule inhibitors and a previously unknown mechanism of ONC201, an anti-cancer small molecule, which led to a successful Phase 2 trial in a rare glioblastoma. Building on these approaches, we also introduce our suite of drug synergy prediction models. These models can not only predict the level of expected drug synergy in specific cancer types, but can also be used to pinpoint the specific mechanisms of actions that contribute to synergy. Altogether, OneThree’s platform is a comprehensive AI approach that combines high quality AI with mechanistic biology to optimize early-stage drug development.

Citation Format: Coryandar Gilvary, Neel Madhukar, Olivier Elemento. OneThree Biotech’s artificial intelligence platform for optimizing drug development [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B031. doi:10.1158/1535-7163.TARG-19-B031
Abstract B037: Potential next-generation androgen receptor-targeted therapeutic for enzalutamide-resistant prostate cancer; In vivo characterization in immune-compromised SRG rats
Authors: Ponnusamy Suriyan Udhayasuriyan , Noto Fallon K , Adedeji Bisoye Towobola , He Yali , Hwang Dong-Jin , Moody Sam , Thiyagarajan Thirumagal , Jamling Tseten Yeshi , Miller Duane D , Narayanan Ramesh
Published: March 2020

Current treatment options for advanced castration-resistant prostate cancers (CRPC) are effective for a brief period before becoming refractory. It is important to use relevant in vivo models for candidate selection in the development of next-generation mechanistically-distinct drugs to treat castration- and drug- resistant prostate cancers. Here, we describe a series of AR pan-antagonists (selective AR degraders (SARDs)) that degrade the AR and AR splice variants. SARDs inhibit the wild-type and LBD mutant ARs comparably and inhibit the in vitro proliferation and in vivo growth of enzalutamide-sensitive and resistant prostate cancer xenografts. Xenograft studies conducted in immune-compromised SRG rats (Sprague Dawley Rag2-/- Il2rg -/-; Hera Biolabs) with three lead SARDs demonstrated regression of enzalutamide-sensitive and -resistant VCaP tumors both in castrated and in intact rats. SRG rats provide the benefit of abundant blood samples for weekly evaluation of rising PSA, which also indicated that the SARDs inhibit the rising PSA, while enzalutamide was inactive in enzalutamide-resistant model. Drug metabolism and pharmacokinetic (DMPK) studies, also conducted with SRG and wild-type rats and in combination with efficacy, indicate that the molecules possess all the necessary drug-like properties. The molecules exhibit a broad safety margin with no cross-reactivity with G-Protein Coupled Receptor, kinase, and nuclear receptor family members. Collectively, the SARDs exhibit the properties necessary for a next-generation prostate cancer drug and that use of SRG rats facilitated thorough characterization of their in vivo properties.

Citation Format: Suriyan Udhayasuriyan Ponnusamy, Fallon K Noto, Bisoye Towobola Adedeji, Yali He, Dong-Jin Hwang, Sam Moody, Thirumagal Thiyagarajan, Tseten Yeshi Jamling, Duane D Miller, Ramesh Narayanan. Potential next-generation androgen receptor-targeted therapeutic for enzalutamide-resistant prostate cancer; In vivo characterization in immune-compromised SRG rats [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B037. doi:10.1158/1535-7163.TARG-19-B037