tbuckley/cabot-search huggingface dataset:
Abstract 1057: Targeting MTHFD2 using first-in-class inhibitors kills haematological and solid cancer through thymineless-induced replication stress
Authors: Thomas Helleday
Journal: Cancer Research (2020)

Abstract Summary Thymidine synthesis pathways are upregulated pathways in cancer. Since the 1940s, targeting nucleotide and folate metabolism to induce thymineless death has remained first-line anti-cancer treatment. Recent discoveries that showing cancer cells have rewired networks and exploit unique enzymes for proliferation, have renewed interest in metabolic pathways. The cancer-specific expression of MTHFD2 has gained wide-spread attention and here we describe an emerging role for MTHFD2 in the DNA damage response (DDR). The folate metabolism enzyme MTHFD2 is one of the most consistently overexpressed metabolic enzymes in cancer and an emerging anticancer target. We show a novel role for MTHFD2 being essential for DNA replication and genomic stability in cancer cells. We describe first-in-class nanomolar MTHFD2 inhibitors (MTHFD2i), with protein co-crystal structures demonstrating binding in the active site of MTHFD2 and engaging with the target in cells and tumours. We show MTHFD2i reduce replication fork speed and induce replication stress, followed by S phase arrest, apoptosis and killing of a range of haematological and solid cancer cells in vitro and in vivo, with a therapeutic window spanning up to four orders of magnitude compared to non-transformed cells. Mechanistically, MTHFD2i prevent thymidine production leading to mis-incorporation of uracil into DNA and replication stress. As MTHFD2 expression is cancer specific there is a potential of MTHFD2i to synergize with other treatments. Here, we show MTHFD2i synergize with dUTPase inhibitors as well as other DDR inhibitors and demonstrate the mechanism of action. These results demonstrate a new link between MTHFD2-dependent cancer metabolism and replication stress that can be exploited therapeutically. Keywords MTHFD2, one-carbon metabolism, folate metabolism, DNA replication, replication stress, synthetic lethal, thymineless death, small-molecule inhibitor, DNA damage response Citation Format: Thomas Helleday. Targeting MTHFD2 using first-in-class inhibitors kills haematological and solid cancer through thymineless-induced replication stress [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1057.
Abstract 5879: Spatially-segmented single-cell transcriptomics by diffusional accessibility to a small-molecule dye
Authors: David B. Morse, Michele Ceribelli, Joachim De Jonghe, Aleksandra Michalowski, Christoph Muus, Maria Vias, Samantha Boyle, David A. Weitz, James Brenton, Jason D. Buenrostro, Craig Thomas, Tuomas P. Knowles
Journal: Cancer Research (2020)

Abstract High-throughput single-cell RNA-seq (scRNA-seq) is used to describe complex tissues by characterizing transcriptional states of individual cells. Defining a cell's position, both in regard to tissue margins and its social context, is essential for understanding the intrinsic and extrinsic variables that effect the transcriptional identity of individual cells. Conventional high-throughput scRNA-seq assays, however, decouple cells from their original locations within tissues. In situ hybridization readouts of gene expression and in situ sequencing preserve spatial information in tissues, but currently have a lower total read threshold than NextGen sequencing – imposing a restriction on either cell throughput or transcriptional breadth. Combining the above methods or using regional barcodes to define 2D positions have spatially-reconstructed tissue regions but are seldom employed in unspecialized laboratories. We describe SEgmentation by Exogenous Perfusion (SEEP), a rapid, integrated, method for providing 3D spatial-segmentation to scRNA-seq data. Tissues are divided into layers based on accessibility of a fluorescent dye allowing sorted cells to be characterized by transcriptomic and regional identity. We use SEEP to explore how the transcriptional states of cells in high-grade serous ovarian cancer vary with respect to intratumoral position. We describe an integrated method for correlating 3D radial-spatial cell positions with scRNA-seq data. By employing a basic stain-and-sort method using an off-the-shelf live-dead stain, we are able to correlate transcriptional profiles with radial-spatial positions in both radially symmetric tissues (e.g., spheroids, organoids, spherical tumor masses) and linear tissue samples (e.g., punch biopsies). SEEP uses an imaging-based calibration step to inform the parameters of a FACS and sequencing based measurement step. Across spheroid, organoid, and PDX models of high-grade serous ovarian cancer, we find common positional transcriptomic profiles, ranging from single-gene proclivities to multi-gene-set enrichments across tissue layers. Citation Format: David B. Morse, Michele Ceribelli, Joachim De Jonghe, Aleksandra Michalowski, Christoph Muus, Maria Vias, Samantha Boyle, David A. Weitz, James Brenton, Jason D. Buenrostro, Craig Thomas, Tuomas P. Knowles. Spatially-segmented single-cell transcriptomics by diffusional accessibility to a small-molecule dye [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 5879.
A Cullin3-KLHL20 Ubiquitin Ligase-Dependent Pathway Targets PML to Amplify HIF-1 Signaling and Tumor Hypoxia responses.
Authors: WC Yuan, YR Lee, SF Huang, YM Lin, TY Chen, HC Chung, CH Tsai, HY Chen, CT Chiang, CK Lai, LT Lu, CH Chen, DL Gu, YS Pu, YS Jou, KP Lu, PW Hsiao, HM Shih, RH Chen
Journal: Cancer Cell (2011)
Abstract 5251: Reproducible, MoA-reflecting reporter-based bioassays to enable drug development of biosimilars and biobetters
Authors: Steven Edenson, Richard Moravec, Dun Li, Jennifer Wilkinson, Frank Fan, Mei Cong
Journal: Cancer Research (2020)

Abstract Cytokines and growth factors are small immunomodulatory proteins secreted by a wide variety of cells including fibroblasts, endothelial and stromal cells, whose role is to regulate surrounding cells in an autocrine, paracrine or endocrine fashion. Immunocytokines represent a promising class of activators of the immune system, with the potential to be used alone or in combination with other therapeutic modalities. Many are currently FDA approved therapy agents (e.g. IFN, IL-2 and Epo), while others are targets for approved biological blocking therapies to treat a variety of diseases. Examples of cytokine blocking agents include basiliximab (IL-2R), tocilizumab and sarilumabad (IL-6R), siltuximab (IL-6), ustekinumab and its biosimilars (IL-12/IL-23 p40), secukinumab (IL-17A), bevasizumab (VEGF), and denosumab (RANKL). Many more are in development and trials as biosimilars and biobetters. IL-2 and IL-15 are still clinically important cytokines as researchers look to improve potency, patient tolerance and response by developing new molecules with sustained and targeted activities. We have developed luciferase reporter bioassays which individually can be used for the quantitation of a variety of cytokines and growth factors including IL-2, IL-6, IL-10, IL-12, IL-15, IL-17, IL-23, BCMA, TGF-b, VEGF and RANKL using respective mechanism of action pathways. The bioassay format is based on thaw-and use cells, eliminating the need to establish and pre-culture cytokine responsive cell lines which provides the benefits of convenience, reproducibility, and transferability. We demonstrate these assays measure cytokine response and inhibition with blocking drugs or potency changes from stressed samples. In summary, these reporter-based bioassays provides valuable tools for the development, stability testing, and potency determination in the manufacture of cytokine biosimilars and biobetters. Citation Format: Steven Edenson, Richard Moravec, Dun Li, Jennifer Wilkinson, Frank Fan, Mei Cong. Reproducible, MoA-reflecting reporter-based bioassays to enable drug development of biosimilars and biobetters [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 5251.
Abstract 464: Allicin, a potent new ornithine decarboxylase inhibitor in neuroblastoma
Authors: Chad R. Schultz, Martin C. Gruhlke, Alan J. Slusarenko, Andre S. Bachmann
Journal: Cancer Research (2020)

Abstract The purpose of this study was to test if allicin inhibits ornithine decarboxylase (ODC) in pediatric neuroblastoma (NB). The natural product allicin is a reactive sulfur species (RSS) from garlic (Allium sativum L.). NB is an early childhood cancer arising from the developing peripheral nervous system. In up to 25% of cases MYCN gene amplification is correlated to high tumor stage and poor patient prognosis. High stage NB relapses frequently despite multimodal therapy and relapsed tumors are virtually untreatable. Therefore, there is a clear need for specific, novel therapeutics. MYCN transactivates the E-box gene ODC1, and the gene product ODC is a rate-limiting enzyme in polyamine biosynthesis. The increase of polyamines (putrescine, spermidine, spermine) triggers cell hyperproliferation in NB and other MYC-driven cancers through the activation of Rb-regulated cell cycle progression. ODC is a validated drug target and α-difluoromethylornithine (DFMO) is an ODC inhibitor under investigation in phase II NB trials. Although a safe drug with clinical promise, DFMO has various challenges including the need for exceptionally high treatment doses and rapid renal clearance in the urine. In an effort to identify pharmacologically superior ODC inhibitors, we identified allicin as a potent ODC inhibitor using a specific radioactive in vitro activity assay with purified ODC. Allicin also inhibited ODC activity in actively growing NB cells, reduced polyamine levels, and induced apoptosis in cell cultures. ODC is a homodimer with 12 cysteines per monomer and allicin S-thioallylates cysteines. Allicin reacts with, and is titrated-out by, low molecular weight thiols like dithiothreitol (DTT). Removal of DTT from the ODC activity assay reaction resulted in significantly higher allicin potency (IC50 at 11 nM). Although allicin has multiple cellular targets, our data reveal that one mode of action is ODC inhibition, which suppresses polyamine accumulation and induces apoptosis. The natural product allicin could be used in conjunction with other anticancer treatments, the latter at a lower than usual dosage, to achieve drug synergism with good prognosis and less side-effects. Citation Format: Chad R. Schultz, Martin C. Gruhlke, Alan J. Slusarenko, Andre S. Bachmann. Allicin, a potent new ornithine decarboxylase inhibitor in neuroblastoma [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 464.
Abstract 4931: Yap1 regulates the transcription of cofilin to facilitate the metastatic movement of colorectal cancer cells
Authors: S M Anisul Islam, Mildred Acevedo-Duncan
Journal: Cancer Research (2020)

Abstract Colorectal cancer (CRC) is one of the most leading death-causing cancers in the United States which accounts for 23,380 estimated death cases in 2019 as per American Cancer Society. Surgical removal of cancerous tissue coupled with chemotherapeutic intervention is the primary treatment of metastatic CRC and the only hope of enhanced survival. Dysregulation of kinases is pivotal in the various pathological process, including cancer. In addition, the metastatic events in a cancer cell are usually brought by the alteration of different key molecules. One such key molecule is called cofilin that acts on the actin cytoskeleton to induce metastatic phenotype in cancerous cells. In this study, we used two atypical Protein Kinase C (PKC) inhibitors, namely ICA-I (a PKC-ι inhibitor) and ζ-Stat (a PKC-ζ inhibitor), to investigate the role of atypical PKCs in regulating the expression of cofilin in metastatic colorectal cells. Our findings suggested that the atypical PKC inhibitors reduced the expression and translocation of Yap1 from cytosol to nucleus which ultimately leads to the reduced expression of cofilin in CRC cells. This mechanistic study provides an important insight into the metastatic behavior of the cancerous colorectal cells that can be used to target metastasis regulating molecules to improve the survival rate in patients with CRC. Citation Format: S M Anisul Islam, Mildred Acevedo-Duncan. Yap1 regulates the transcription of cofilin to facilitate the metastatic movement of colorectal 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 4931.
IL-10 production by ILC2s requires Blimp-1 and cMaf, modulates cellular metabolism, and ameliorates airway hyperreactivity
Authors: Emily Howard, Gavin Lewis, Lauriane Galle-Treger, Benjamin P. Hurrell, Doumet Georges Helou, Pedram Shafiei-Jahani, Jacob D. Painter, German Aleman Muench, Pejman Soroosh, Omid Akbari
Journal: Journal of Allergy and Clinical Immunology (2020)
Abstract 3958: Establishment of an <i>in vitro</i> model to study cancer associated fibroblasts and their direct effect on desmoplasia
Authors: Neel Ingemann Nissen, Morten Karsdal, Nicholas Willumsen
Journal: Cancer Research (2020)

Abstract Background: The extracellular matrix (ECM) and desmoplasia is emerging as key components in defining prognosis and response to treatment. Cancer associated fibroblasts (CAFs) are vital contributors to the desmoplastic reaction and novel ambitious therapies targeting tumor specific ECM and CAFs are being developed. However, it is still debated whether in fact desmoplasia acts as a barrier to- or promoter of - tumor growth. Therefore, there is a need to establish novel models of desmoplasia to improve the understanding of CAFs and associated cancer specific ECM alterations in order to be successful in drug development and prediction of treatment success. By applying the “Scar-in-a-jar” (SiaJ) model, we established an in vitro model including healthy fibroblasts and CAFs derived from the pancreas to study ECM changes and desmoplasia. Methods: Primary healthy fibroblasts and CAFs were cultured for up to 12 days in the presence or absence of TGF-β. Type I (PRO-C1), type III (PRO-C3) and type VI (PRO-C6) collagen formation were quantified in cell supernatant using enzyme-linked immunosorbent assays. Metabolic activity was evaluated by AlamarBlue. Results: In healthy fibroblasts no collagen formation could be detected without TGF-β. However, the presence of TGF-β induced PRO-C1, PRO-C3 and PRO-C6 formation with levels increasing up to 20-fold (p&amp;lt;0.0001). Furthermore, PRO-C1, PRO-C3 and PRO-C6 levels where significantly increased in CAFs compared to healthy fibroblasts independent of TGF-β stimulation. There was no difference in metabolic activity across cell line. Conclusions: TGF-β stimulation led to increased ECM protein expression in healthy fibroblasts but not in CAFs. However, CAFs expressed higher levels of collagens than healthy fibroblasts independent of TGF-β. These results suggest a distinct role of CAFs in their contribution to desmoplasia compared to healthy fibroblasts. The “Scar-in-a-Jar” model can provide a novel in vitro tool to study CAF activity and their direct effect on desmoplasia. Citation Format: Neel Ingemann Nissen, Morten Karsdal, Nicholas Willumsen. Establishment of an in vitro model to study cancer associated fibroblasts and their direct effect on desmoplasia [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 3958.
Abstract 4188: The ERK1/2 inhibitor, JSI-1187, demonstrates preclinical efficacy in tumor models with MAPK pathway mutations
Authors: Ao Li, Shanzhong Jian, Xia Yuan, Fengge Song, Shengwei Yang, Chengyi Du, Yue Tao, Liucheng Wang, Mingming Pan, Penghui Dong, jiajun Zhou, Zhengkang Ge, Qinghong Zhu, Wenshan Hao, Wen Xu, Jintao Zhang, Qun Li, Shaohui Wang
Journal: Cancer Research (2020)

Abstract The MAPK pathway, via RAS-RAF-MEK-ERK, is one of the most important pathways for relaying signals from cell surface receptors for cell survival, proliferation, and differentiation. Many mutations were identified in this pathway and intensely studied, resulting in a number of therapeutics for various cancers (for example, EGFR mutant inhibitors, BRAF V600 mutant inhibitors and MEK inhibitors). A common mechanism of resistance to these therapeutics has emerged to be the reactivation of ERK1/2, the terminal kinase in this pathway. Inhibition of ERK1/2 activity could therefore be instrumental in various mutations in the MAPK pathway, either as single therapy or in combination with other compounds. Through internal effort, we discovered a novel ERK1/2 small molecule inhibitor, JSI-1187, which reversibly inhibited ERK2 with a Ki of less than 1 nM. In anti-proliferation assays, JSI-1187 had IC50 of 37 NM in the BRAF V600E melanonma cell line A375 and 48 NM in the BRAF V600E CRC cell line COLO 205. For cell lines with KRAS mutations, JSI-1187 had anti-proliferation IC50 of 130 NM in the CRC cell line HCT116 and 150 NM in the NSCLC cell line Calu-6. Furthermore, JSI-1187 inhibited the phosphorylation of the ERK substrate RSK1 in these cell lines. We also tested JSI-1187 using mouse CDX models. JSI-1187 showed tumor growth inhibition in a dose dependent manner in the xenograft models of A375, COLO 205, HCT116, and Calu-6. In the COLO 205 model, JSI-1187 caused tumor regression at the dose of 20 mg/kg QD. In the Calu-6 model, BID and QD dosing schedules were compared, with the result that 10 mg/kg BID and 20 mg/kg QD inhibit xenograft tumor growth similarly. Moreover, in the HCT116 model, JSI-1187 concentrations in plasma and tumor were measured, as well as phopspho-RSK1 levels in tumor. A PK/PD correlation was observed. In addition to being tested alone, JSI-1187 was also tested in combination with the BRAF inhibitor dabrafenib in the COLO 205 xenograft model. JSI-1187 and dabrafenib demonstrated synergistic efficacy in tumor growth inhibition. In conclusion, JSI-1187 can be potentially used for treatment of cancer with MAPK pathway mutations. Citation Format: Ao Li, Shanzhong Jian, Xia Yuan, Fengge Song, Shengwei Yang, Chengyi Du, Yue Tao, Liucheng Wang, Mingming Pan, Penghui Dong, jiajun Zhou, Zhengkang Ge, Qinghong Zhu, Wenshan Hao, Wen Xu, Jintao Zhang, Qun Li, Shaohui Wang. The ERK1/2 inhibitor, JSI-1187, demonstrates preclinical efficacy in tumor models with MAPK pathway mutations [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 4188.
Abstract 1543: Development of a novel ECAD-EmGFP reporter line for pancreatic cancer MET study and drug discovery
Authors: Diana Douglas, Sangeeta Kumari, Jarkko Huuskonen, Weiguo Shu
Journal: Cancer Research (2020)

Abstract World-wide, metastasis continues to be the leading cause of death in cancer patients1. Although epithelial-to-mesenchymal transition (EMT) and mesenchymal-to-epithelial transition (MET) have been implicated in the progression of cancer metastasis and drug resistance, their mechanistic pathway and specific role in disease progression is not fully understood2. As such, the development of a reporter line that enables the real-time monitoring of the changing status of cells will not only aid in dissecting the EMT/MET pathway in the research field, but could also become a robust platform for new cancer drug development. E-cadherin, an adhesion protein expressed in epithelial cells, is upregulated in pancreatic cancer cells during MET and is associated with an increase in tight junctions and apico-basal polarity, as well as a change in morphology3,4. Here, we developed a novel PANC-1 ECAD-EmGFP reporter line using CRISPR/Cas9 genome-editing technology. In this cell line, the emerald green fluorescent protein (EmGFP) reporter was incorporated into the last exon of the endogenous E-cadherin gene, enabling real-time monitoring of MET status in live cells. The ECAD-EmGFP knock-in allele was confirmed at genomic, transcriptional, and translational levels. Functional data revealed that miRNA-200 treatment induced the increased expression of ECAD-EmGFP, and decreased expression of Snail, a transcription factor associated with mesenchymal traits. These changes in marker gene expression, as well as the decrease in invasive capacity upon induction, suggest that cells have undergone MET. This cell line is a valuable tool for dissecting the molecular mechanisms underlying EMT and MET and for evaluating or screening compounds targeting EMT and MET in pancreatic cancer. 1. Dizon DS, et al. Clinical Cancer Advances 2016: Annual Report on Progress Against Cancer from the American Society of Clinical Oncology. J Clin Oncol 34: 987-1011, 2016. 2. Nieto MA, et al. EMT: 2016. Cell 166: 21-45, 2016. 3. Polireddy K, et al, Targeting Epithelial-Mesenchymal Transition for Identification of Inhibitors for Pancreatic Cancer Cell Invasion and Tumor Spheres Formation. PLOS ONE 11(10): e0164811, 2016 4. Wong S, et al. E-cadherin: Its dysregulation in carcinogenesis and clinical implications. Critical Reviews in Oncology / Hematology 121: 11- 22, 2018 Citation Format: Diana Douglas, Sangeeta Kumari, Jarkko Huuskonen, Weiguo Shu. Development of a novel ECAD-EmGFP reporter line for pancreatic cancer MET study and 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 1543.