bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 894: A Self-Immobilizing NIR Probe for in vivoImaging of Chemotherapy Induced Senescence
Authors: Ma Xiaowei , Liu Jun , Wang Ying , Cui Chao , Deenik Philip
Published: August 2019

Post-treatment relapse and distant metastasis are significant causes of mortality by cancers. Previous research has demonstrated that cancer treatments, such as chemotherapy, can sometimes stimulate tumor spread. This phenomenon is associated with an essential process that some cells are resistant to apoptosis by entering premature senescence after therapy. Thus, chemotherapy-induced cellular senescence may play a critical role in cancer recurrence and metastasis. To monitor the cellular senescence induced by chemotherapy, we developed a self-immobilizing near infrared (NIR) probe (4-152) that can be activated by senescence-associated beta-galactosidase (β-gal) and get immobilized at site of activation, allowing the monitoring of chemotherapy-induced senescence in vivo. We first verified our approach in vitro, when cervical cancer cells (HeLa) were treated with camptothecin (CPT), followed by fluorescence imaging with 4-152. The results demonstrated that our designed probe exhibited a dramatic NIR fluorescent response in cells receiving CPT treatment. The fluorescent western blot showed that the probe could be activated by β-gal and attached to the enzyme. The confocal microscopy imaging and flow cytometry results further demonstrated the 4-152 could accumulate in the CPT-induced senescent cancer cells. We then investigated the in vivo imaging ability of the probe for senescence detection in tumor xenografts treated with CPT. After treatment with CPT or saline (as control), 4-152 was administrated to the mice by tail vein injection. NIR imaging was then performed on the IVIS Spectrum in vivo imaging system. The imaging showed that the 4-152 could quickly distribute to the tumor and accumulated for over 24 hours. The fluorescence intensity of the CPT treated tumors was significantly higher than the untreatment tumors. After imaging, tumors and major organs were resected for ex vivo imaging. The fluorescent intensity of tumors showed 6.27-fold higher than liver. Ex vivo immunofluorescence staining and X-gal staining further confirmed the correlation of probe 4-150 and senescence cells.In conclusion, we have developed an activatable self-immobilizing NIR probe for the monitoring of chemotherapy-induced cellular senescence in vivo. This probe may be used for the treatment effect monitoring and therapeutic regimen optimization in the future.

Reference1. Ebos JM. Prodding the beast: assessing the impact of treatment-induced metastasis. Cancer Res 2015; 75: 3427-3435.

Citation Format: Xiaowei Ma, Jun Liu, Ying Wang, Chao Cui, Philip Deenik. A Self-Immobilizing NIR Probe for in vivoImaging of Chemotherapy Induced Senescence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 894.
Abstract 897: Lipid metabolism is involved in mitotic slippage-induced SASP upon treatment with anti-mitotic drugs
Authors: Crasta Karen C. , Wong Alex , Lim Bryan
Published: August 2019

Aberrant lipid accumulation is a hallmark of cancer. However, there is little known regarding mechanisms linking lipid metabolism to chemotherapy-induced cell fates. Here, we describe lipid accumulation in cells following antimitotic drug-treatment. Cells arrested in mitosis, as well as cells that escaped mitotic arrest and underwent mitotic slippage, showed elevated cytoplasmic lipid droplets. Interestingly, we found that TOFA, an inhibitor that targets acetyl-CoA carboxylase (ACC) and blocks lipid accumulation enhanced survival of antimitotic-treated cells. Our work previously revealed that cells that survive slippage become senescent and confer pro-tumourigenic effects through non-cell-autonomous effects. Modulating lipid biosynthesis in cells post-slippage by TOFA amplified their inflammatory secretion profiles and accelerated development of tumourigenic behavior. In contrast to TOFA, inhibition of lipid accumulation by C75, a drug targeting fatty acid synthase, significantly reduced the production of pro-tumourigenic factors. This suggests that discrete lipid biosynthesis pathways could contribute differentially to the regulation of pro-tumourigenic inflammation.

Note: This abstract was not presented at the meeting.

Citation Format: Karen C. Crasta, Alex Wong, Bryan Lim. Lipid metabolism is involved in mitotic slippage-induced SASP upon treatment with anti-mitotic drugs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 897.
Abstract 911: UCSC Xena for cancer genomics visualization and interpretation
Authors: Goldman Mary , Craft Brian , Zhu Jingchun , Haussler David
Published: August 2019

UCSC Xena (http://xena.ucsc.edu/) is a web-based visual integration and exploration tool for multi-omic data and associated clinical and phenotypic annotations. Our unique Visual Spreadsheet shows multiple data types side-by-side enabling discovery of correlations across and within genes and genomic regions. We offer dynamic Kaplan-Meier survival analysis, powerful filtering and subgrouping, charts, statistical analyses, genomic signatures, comparative transcript views, and bookmarks. We link out to the UCSC Genome Browser, giving users additional genomic context for any gene or coordinate, as well as MuPIT/CRAVAT and TumorMap, to give users complementary views of the same data.

Xena showcases seminal cancer genomics datasets from TCGA, the Pan-Cancer Atlas, PCAWG, GDC, GTEx, ICGC, and more; a total of more than 1500 datasets across 50 cancer types. We support virtually any type of functional genomics data. In addition to the commonly available SNPs, INDELs, copy number variation, and gene expression datasets, we support DNA methylation, exon-, transcript-, miRNA-, lncRNA-expression and structural variants. We also support clinical data such as phenotypes, subtype classifications and biomarkers. A recompute of TCGA, TARGET and GTEx datasets through the same bioinformatics pipeline allows users to compare expression between tumor and normal tissues. All of our data is available for download via our python API or through AWS S3 buckets.

A researcher can host their own data securely via private hubs running on a laptop or behind a firewall, with visual and analytical integration occurring only within the Xena Browser. The lightweight Xena Data Hubs are straightforward to install on Windows, Mac and Linux. Loading data is easy using either our application or command line interface.

Our newest features include:

* a new, more intuitive wizard to load your data into a local Hub

* URL bookmarks to save interactive views for yourself or to share with collaborators

* genomic signatures: dynamically build as a weighted sum over a set of genes

* hierarchically cluster a list of genes, regulons or probes

* upper vs lower quartile in a KM plot

Citation Format: Mary Goldman, Brian Craft, Jingchun Zhu, David Haussler. UCSC Xena for cancer genomics visualization and interpretation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 911.
Abstract 937: Nr4a transcription factors limit CAR T-cell function in solid tumors
Authors: Chen Joyce , Lopez-Moyado Isaac F. , Seo Hyungseok , Lio Chan-Wang J. , Hempleman Laura J. , Sekiya Takashi , Yoshimura Akihiko , Scott-Browne James P. , Rao Anjana
Published: August 2019

In cancer immunotherapy, CD19-targeted CAR T cells have exhibited impressive clinical efficacy against B cell leukemias and lymphomas; however, they have been less effective against solid tumors. This is in part because CAR T cells enter a hyporesponsive (“exhausted” or “dysfunctional”) state that is triggered by chronic antigen stimulation and characterized by upregulation of several inhibitory surface receptors and loss of effector function. To identify transcriptional regulators and other candidates contributing to the diminished CAR T cell function in solid tumors, we developed a CAR T cell model in which recipient mice bearing murine melanoma tumors expressing human CD19 were adoptively transferred with CD19-targeted CAR T cells. We demonstrate that in both CD8+ CAR T tumor-infiltrating lymphocytes (TILs) and endogenous CD8+ TILs expressing high levels of PD-1 and TIM3, the Nr4a proteins Nr4a1 (Nur77), Nr4a2 (Nurr1), and Nr4a3 (Nor1) are prominent effectors of the transcriptional program downstream of NFAT: they promote the expression of inhibitory receptors and genes associated with early hyporesponsive state, and limit effector function. Most importantly, treatment of tumor-bearing mice with CD8+ CAR T cells lacking all three Nr4a factors (Nr4aTKO) resulted in tumor regression and prolonged survival. Nr4aTKO CD8+ CAR T TILs displayed a transcriptomic profile characteristic of effector function, including upregulation of granzymes and cytokines, and many of these gene expression changes were associated with altered regulatory element accessibility near effector genes. Our data identify Nr4a transcription factors as major players in the cell-intrinsic program of T cell hyporesponsiveness and point to Nr4a inhibition as a promising strategy for cancer immunotherapy.

Citation Format: Joyce Chen, Isaac F. Lopez-Moyado, Hyungseok Seo, Chan-Wang J. Lio, Laura J. Hempleman, Takashi Sekiya, Akihiko Yoshimura, James P. Scott-Browne, Anjana Rao. Nr4a transcription factors limit CAR T-cell function in solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 937.
Abstract 938: Disruption of TOX overcomes CAR T-cell dysfunction function in solid tumor
Authors: Seo Hyungseok , Chen Joyce , Das Arundhoti , Bhandoola Avinash , Rao Anjana
Published: August 2019

Adoptive cell therapy with chimeric antigen receptor expressing T (CAR T) cells has shown promising therapeutic efficacy against leukemia and lymphoma. However, CAR T cells in solid tumors fail to be as effective as in liquid tumors since they enter into a hyporesponsive (exhausted or dysfunctional) state that is induced by chronic antigen stimulation in cancer. Here, we show that CAR T cells in solid tumors exhibit low effector function and high expression of inhibitory receptors such as PD-1, Tim-3 and Lag 3. We also show that the nuclear factor, Thymocyte selection-associated HMG box protein, TOX - a target of the transcription factor NFAT - is highly expressed in CAR T cells in solid tumors with PD-1 expression. To determine how TOX transcription factors contribute to exhaustion/ dysfunction in CAR T cells, we developed CAR T cells targeting huCD19 and lacking TOX and TOX 2. Surprisingly, treatment of huCD19 expressing melanoma or colorectal tumor-bearing mice with CAR T cells doubly deficient in TOX and TOX2 resulted in the eradication of solid tumors and prolonged survival of the tumor-bearing mice, compared to mice treated with WT CAR T cells or CAR T cells lacking TOX or TOX2 alone. Tumor-infiltrating CAR T cells (CAR TILs) doubly deficient in TOX and TOX2 did not increase their expression of inhibitory receptors such as PD-1, Lag3 and CD160 and showed higher expression of TCF1 compared to WT CAR TILs. Our data identify TOX and TOX2 -high mobility group transcription factors belonging to the TOX family - as critical transcriptional factors for inducing hyporesponsiveness in CAR T cells in solid tumors. Our data suggest that disruption of TOX expression or activity could be a promising strategy for cancer immunotherapy.

Citation Format: Hyungseok Seo, Joyce Chen, Arundhoti Das, Avinash Bhandoola, Anjana Rao. Disruption of TOX overcomes CAR T-cell dysfunction function in solid tumor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 938.
Abstract 968: Targeting intratumoral heterogeneity in TNBC with novel combinations of FDA-approved drugs
Authors: Truchan Nathan A. , Burnett Joseph P. , Sun Duxin
Published: August 2019

Triple negative breast cancer, like so many other solid tumors, has intratumoral heterogeneity which makes it difficult to treat. It is paramount to understand the question, “where does the intratumoral heterogeneity come from?” To that end, we developed a dual color reporter cell line using CRISPR/Cas9 to precisely knock in fluorescent proteins, mCherry and EGFP, behind CSC marker ALDH1A3 and differentiation marker CD24 respectively, in the TNBC cell line SUM149. With fluorescent microscopy we are able to track individual cells over time, view symmetrical and asymmetrical self-renewal, differentiation, and lineage restriction. Through direct observation we determined that the intratumoral heterogeneity comes from CSCs. In short, CSCs give rise to all the different cell types found within TNBC. Once we established the reporter cell line we wanted to ensure that any response to therapeutics was the same as in the parental SUM149 cell line. In the past, we have treated parental SUM149 with docetaxel or sulforaphane. We therefore treated our reporter cells with either docetaxel or sulforaphane. The sulforaphane treatment resulted in a significant reduction of CSCs while the docetaxel resulted in significantly more CSCs, both results matched the results from parental SUM149. To our surprise the docetaxel treatment result was due to the significant de-differentiation of mCherrylow to mCherryhigh cells. The de-differentiation and incomplete elimination of the heterogeneous cell populations pose a significant problem for TNBC patients, one that must be addressed. In order to overcome this issue, we will use our reporter cells to test already approved FDA drugs to find and evaluate novel combinations for their ability to completely eliminate the heterogeneous cell populations found within TNBC.

Citation Format: Nathan A. Truchan, Joseph P. Burnett, Duxin Sun. Targeting intratumoral heterogeneity in TNBC with novel combinations of FDA-approved drugs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 968.
Abstract 985: Development and validation of cell-based TFEB translocation assay in a high-content and high-throughput screening format
Authors: Zhang Li , Huang Ruili , Ding Wen Xing , Xia Menghang
Published: August 2019

Transcription factor EB (TFEB) is a master regulator of lysosomal gene expression and mitochondrial biogenesis. Autophagy is known to act as a double edged sword in cancer. Autophagy acts as a tumor suppressor during the tumorigenesis process but it also helps to sustain the survival of already transformed cancer cells. Lysosomes sit at the end of the autophagy pathway, which is critical to meet the needs of autophagy. Therefore, small molecules that inhibit TFEB-mediated lysosomal biogenesis and autophagic degradation may be helpful for inhibiting cancer cell growth and limiting cancer progression. On the other hand, small molecules that enhance TFEB-mediated lysosomal degradation may be used to prevent tumorigenesis. In order to identify compounds that modulate TFEB translocation, an image-based TFEB translocation assay was developed in a 1536-well plate format with a GFP-TFEB stable AML12 cell line, which is an immortalized mouse hepatocyte cell line. Torin 1, a known TFEB translocation inducer, was used as positive control in the study. Torin 1 increased TFEB nuclear translocation in a concentration dependent manner with an EC50 of 150nM. To optimize the assay condition, various cell densities per well and time courses were tested. To validate this assay, a group of 384 known anticancer compounds were screened at 11 concentrations ranging from 0.8 nM to 46 μM. From this screening, we identified 25 compounds that enhanced TFEB nuclear translocation with EC50s ranging from 2 nM to 12 μM. Several known and novel TFEB inducers were among these compounds. These results demonstrate that this high-content and high-throughput assay can be used to screen large numbers of chemicals and provide a robust in vitro system to identify TFEB agonists and antagonists, which may be used to either prevent (agonists) or treat (antagonist) cancer.

Citation Format: Li Zhang, Ruili Huang, Wen Xing Ding, Menghang Xia. Development and validation of cell-based TFEB translocation assay in a high-content and high-throughput screening format [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 985.
Abstract 989: The relationship between TP53 gene status and carboxylesterase 2 expression in human gastric cancer
Authors: Kohira Yoshinori , Lee Hyeon-Cheol , Ishimine Momoko , Orita Hajime , Kobayashi Toshiyuki , Sato Koichi , Yokomizo Takehiko , Fukunaga Tetsu
Published: August 2019

Carboxylesterases are serine hydrolases that are involved in the metabolisms of various endogenous and exogenous compounds. They are also required for activation of many anti-cancer prodrugs. For example, irinotecan (CPT-11), an anti-cancer prodrug that has been approved for the treatment of many types of solid tumors including gastric cancer, is converted by the carboxylesterase CES2 to its active metabolite 7-ethyl-10-hydroxycamptothesin (SN-38), a very potent topoisomerase I inhibitor. Among carboxylesterase isozymes, CES2 is most highly expressed in the gastrointestinal tract. Thus, the expression of CES2 may play an important role in local (i.e., intratumoral) activation of anti-cancer prodrugs such as irinotecan in the gut. Recent studies with cultured cancer cell lines have shown that CES2 expression is regulated by the tumor suppressor protein p53. However, whether CES2 expression is affected by the presence of p53 mutation in clinical cancer samples still remains unclear. In this study, we focused on the regulatory mechanism of CES2 expression in gastric cancer. First, we examined the relationship between TP53 gene status and CES2 expression using gastric cancer cell lines. Several gastric cancer cell lines expressing wild-type p53 (AGS, NUGC4, MKN74, and HSC58) were treated with nutlin-3a, a drug that inhibits the interaction between p53 and the E3 ubiquitin ligase MDM2 and thereby directly activates p53 signaling without genotoxic side effects. The expression of p21, a downstream target of p53, was increased following nutlin-3a treatment in two p53 wild-type cell lines NUGC4 and HSC58. The expression of CES2 was also upregulated by nutlin-3a in three p53 wild-type cell lines AGS, NUGC4, and HSC58. As expected, the expression levels of p21 and CES2 were not largely affected by nutlin-3a in gastric cancer cell lines with TP53 mutations. These results indicate that CES2 expression is positively regulated by the p53 pathway in most gastric cancer cells. We also investigated the relationship between TP53 gene status and CES2 expression in human gastric cancer samples. Our results may provide useful information for predicting the efficacy of anti-cancer prodrugs that are activated by CES2 in gastric cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Yoshinori Kohira, Hyeon-Cheol Lee, Momoko Ishimine, Hajime Orita, Toshiyuki Kobayashi, Koichi Sato, Takehiko Yokomizo, Tetsu Fukunaga. The relationship between TP53 gene status and carboxylesterase 2 expression in human gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 989.
Abstract 994: Combining ruthenium metallo-intercalators and targeted radionuclide therapy for EGFR-overexpressing oesophageal cancer
Authors: Gill Martin R. , Menon Jyothi U. , Carlisle Robert , Vallis Katherine A.
Published: August 2019

Targeted radionuclide therapy (TRT) combines the specificity of molecular targeting with the potent cytotoxicity of ionising radiation. An emerging question is how to most effectively integrate TRT with radiosensitizing small molecules [1]. Auger electron-emitting radionuclides such as 111In are well-suited as the therapeutic radionuclide as their short-range reduces “cross fire” damage to neighbouring cells. As a novel chemical class of radiosensitizing agent, we have found ruthenium polypyridyl complexes (RPCs) that intercalate between DNA base pairs such as Ru(phen)2(tpphz)2+ (phen = 1,10 phenanthroline, tpphz = tetrapyridophenazine) act to stall replication forks and impede mitotic progression in human oesophageal cancer cells [2,3]. This dual mechanism of action results in preferential growth inhibition of rapidly-proliferating oesophageal cancer cells with elevated mitotic indices. In contrast to cisplatin and other DNA-damaging agents, relatively low DSB levels are generated in response to RPC metallo-intercalation and reduced cytotoxicity towards non-cancer cells is observed. In addition to this, treatment is able to enhance cancer cell sensitivity to IR by increasing IR-induced DNA double-strand break (DSB) damage, decreasing cell survival in a synergistic manner. Based on these discoveries, we explore how nanoparticle-mediated delivery of Ru(phen)2(tpphz)2+ and the Auger electron emitting radionuclide 111In can achieve localised cytotoxicity enhancement specifically in EGFR-overexpressing oesophageal cancer cells, thereby demonstrating both a combinational effect alongside molecular targeting for this novel combination [4].

References:[1] M. R. Gill, N. Falzone, Y. Du and K. A. Vallis, Lancet Oncol, 18, e414 (2017).[2] M. R. Gill, S. N. Harun, S. Halder, R. A. Boghozian, K. Ramadan, H. Ahmad and K. A. Vallis, Sci. Rep., 6, 31973 (2016).[3] M. R. Gill, P. J. Jarman, S. Halder, M. G. Walker, H. K. Saeed, J. A. Thomas, C. Smythe, K. Ramadan and K. A. Vallis, Chem. Sci., 9, 841 (2018).[4] M. R. Gill, J. U. Menon, P. J. Jarman, J. Owen, I. Skaripa-Koukelli, S. Able, J. A. Thomas, R. Carlisle, K. A. Vallis, Nanoscale, 10, 10596 (2018).

Citation Format: Martin R. Gill, Jyothi U. Menon, Robert Carlisle, Katherine A. Vallis. Combining ruthenium metallo-intercalators and targeted radionuclide therapy for EGFR-overexpressing oesophageal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 994.
Abstract 997: Three-dimensional high-throughput drug screening for bladder cancer
Authors: Lee Sang-Cheol , Kim Young Saing , Hwang In Gyu , Lee Su Jin , Park Se Hoon
Published: August 2019

Recent progress in understanding the molecular mechanisms that underlie bladder cancer carcinogenesis offers the prospect of specifically targeting signaling pathways to achieve more effective and rational treatment. In a novel, 3-dimensional high-throughput drug screening (3D HTS) platform to identify novel agents affecting bladder cancer cells, 24 molecularly-targeted agents were screened in seven human bladder cancer cell lines (UCUM3, T24, SW780, RT4, J82, 5637 and 253J-BV). The efficacy of 24 targeted agents on the bladder cell lines varied dramatically on their genomic alterations. BEZ235 (mTOR and PI3K inhibitor) showed antitumor effect in most of cell lines except UMUC3. Another mTOR inhibitor, AZD2014 (inhibitors of mTORC1 and mTORC2) also had IC50 <2 μM in 3 cell lines (5637, J82 and RT4). However, FGFR targeted agents (AZD4547 and BGJ398) did not show the efficacy in cell lines with FGFR3 alterations (J82, SW780 and RT4). In HER2 mutated/amplified cell line (5637), dacomitinib, lapatinib and neratinib showed anti-tumor effect. Immunoblot assay showed that lapatinib and neratinib suppressed phosphorylated HER2 and downstream molecules such as p-AKT or p-ERK. In conclusion, 3D HTS drug screening platform could be a useful preclinical tool to evaluate various kinds of drugs rapidly, and eventually provide a valuable insight into precision medicine along with comprehensive genomic analysis.

Note: This abstract was not presented at the meeting.

Citation Format: Sang-Cheol Lee, Young Saing Kim, In Gyu Hwang, Su Jin Lee, Se Hoon Park. Three-dimensional high-throughput drug screening for bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 997.
Abstract 998: HEx: A computational and synthetic biology platform applied to oncology drug discovery
Authors: Schlecht Ulrich , Cordero Pablo , Hillenmeyer Maureen , Jeon Ju Eun , Kvitek Dan , Naughton Brian , Wiemann Philipp , Harvey Colin
Published: August 2019

We present a computational and synthetic biology platform for producing novel compounds targeting proteins relevant to oncology. As eukaryotes, fungi share approximately one third of human genes, including many relevant to core metabolism, cell cycle, DNA repair, and protein degradation. We have developed a computational method for mining fungal genomes to identify genes likely to produce compounds that inhibit specific human target proteins, including undruggable targets such as protein-protein interactions. We engineer these genes using our synthetic biology pipeline to produce the novel compounds in a heterologous host. We present in vitro data for an Aurora Kinase (AURKA) inhibitor developed using this platform as proof of concept for Hexagon Bio's pipeline. These data demonstrate the potential of this system to target the more than 6000 genes shared between fungi and humans.

Citation Format: Ulrich Schlecht, Pablo Cordero, Maureen Hillenmeyer, Ju Eun Jeon, Dan Kvitek, Brian Naughton, Philipp Wiemann, Colin Harvey. HEx: A computational and synthetic biology platform applied to oncology drug discovery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 998.
Abstract CT008: Tumor Treating Fields combined with radiotherapy and temzolomide for newly diagnosed glioblastoma: Final results from a pilot study
Authors: Grossman Rachel , Limon Dror , Bokstein Felix , Ram Zvi
Published: August 2019

Background: Tumor Treating Fields (TTFields) are a non-invasive, loco-regional, anti-mitotic treatment comprising low intensity alternating electric fields. In the Phase III EF-14 study in newly diagnosed glioblastoma (ndGBM), TTFields significantly improved survival compared to temozolomide (TMZ). Preclinical data have demonstrated that TTFields increased glioma cell death following radiation therapy (RT), suggesting a radio-sensitizing effect of TTFields. We therefore initiated a pilot study to evaluate the safety and feasibility of administering TTFields concomitant to RT and TMZ in ndGBM patients.

Methods: Patients with histologically confirmed ndGBM were treated with TTFields/RT/TMZ followed by maintenance TTFields and TMZ for up to 24 months. TTFields (200kHz) were delivered for >18 hours/day with removal of the transducer arrays during delivery of RT. TMZ was administered at a dose of 75 mg/m2/daily for 6 weeks and RT at a total dose of 60 Gy. The primary endpoint was safety of the combined TTFields/RT/TMZ; secondary endpoints included progression-free survival (PFS), overall survival (OS) and toxicity. Adverse events (AEs) were graded according to CTCAE V4.0.

Results: Ten patients were enrolled at a single center in Israel between April and December 2017. All patients had recovered from maximal debulking surgery or biopsy. Five patients (50%) had undergone gross total resection while the rest had biopsy only. Median age was 59 and median KPS was 80. Median dose of RT was 60 Gy. Six patients (60%) reported at least one AE. The most common AE was TTFields-related skin toxicity, reported in 4 patients (40%), all of which were Grade 1-2 in severity. Two patients reported serious AEs (seizures and general deterioration) that were considered unrelated to TTFields. Median PFS with RT/TMZ/TTFields was 10.5 months. Median OS has not yet been reached.

Conclusions: The proportion of patients with TTFields-related skin toxicity was similar to that reported in ndGBM patients in the randomized Phase III study (52%). No other TTFields-related toxicities were reported, nor was there an increase in RT- or TMZ-related toxicities as a result of combining TTFields with RT in addition to TMZ. Based on the safety and preliminary efficacy results of this pilot study, a Phase II randomized study has been initiated to investigate the efficacy of concomitant RT/TMZ/TTFields in 60 ndGBM patients.

Citation Format: Rachel Grossman, Dror Limon, Felix Bokstein, Zvi Ram. Tumor Treating Fields combined with radiotherapy and temzolomide for newly diagnosed glioblastoma: Final results from a pilot study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT008.
Abstract CT054: Clinical and pharmacokinetic study with a lipidic prodrug of mitomycin-c entrapped in liposomes in metastatic colorectal cancer patients
Authors: Gabizon Alberto A. , Tahover Esther , Perets Ruth , Golan Talia , Geva Ravit , Amitay Yasmine , Shmeeda Hilary , Ohana Patricia
Published: August 2019

Background: A phase 1A/1B study was conducted to investigate the safety of Promitil, a liposomal formulation of a lipidic prodrug of mitomycin-c (MLP) within an open, dose-escalating, phase (1A, n=27) followed by an expanded phase (1B, n=61) restricted to metastatic colorectal cancer (mCRC) patients after failure to two or more lines of therapy. We report here on the correlation between pharmacokinetic (PK) parameters and clinical observations in patients with mCRC.

Methods: PK analysis of plasma MLP levels was obtained in 53 mCRC patients (F=26, M=27), who received Promitil every 4 weeks either as single agent (n=28), in combination with capecitabine (Cap) (n=12), or in combination with Cap and bevacizumab (Bev) (n=13). The MLP dose range was 0.5-3.5 mg/kg with most patients (N=29) receiving 2.0-2.5 mg/kg. Out of 53 patients analyzed, 39 patients completed the 3rd cycle (study week 12) and were efficacy-evaluable by CT scan and Recist criteria. PK of plasma MLP levels was analyzed by noncompartmental methods. For statistical analysis and correlations of T½ and Clearance (CL) with age, sex, BMI, baseline neutrophil/lymphocyte ratio (NLR), CEA, and survival, we used t test and Pearson or Spearman coefficients.

Results: The PK of MLP was stealth-like with long T½ (~1 day), slow CL, and small Volume of distribution. Stable Disease (SD) was reported in 14 patients (36%), and Progressive Disease (PD) in 25 patients. Median survival of all 39 patients (SD+PD) was 8.3 months. SD patients had significantly longer median survival than PD patients (14.3 vs 6.5 months, p=0.0002). SD patients had significantly longer T½ and slower CL than non-SD patients. A longer T½ and slower CL were significantly correlated with longer survival. A high NLR and high CEA were correlated with shorter T½ and/or faster CL. Dose of MLP, addition of Cap and/or Bev, age and BMI did not have any apparent effect on response or survival.

Conclusions: SD was indicative of extended survival. The longer circulation time of the liposomal prodrug in SD patients and its correlation with longer survival are consistent with the well-known correlation between liposome circulation time and localization in tumors. A high NLR may signal inflammatory macrophage activation resulting in shorter liposomal circulation half-life and reducing the chances of disease control.

Citation Format: Alberto A. Gabizon, Esther Tahover, Ruth Perets, Talia Golan, Ravit Geva, Yasmine Amitay, Hilary Shmeeda, Patricia Ohana. Clinical and pharmacokinetic study with a lipidic prodrug of mitomycin-c entrapped in liposomes in metastatic colorectal cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT054.
Abstract CT055: Microdose induced oxaliplatin-DNA adducts as a predictive biomarker of response in colorectal cancer
Authors: Chen Justin , Cho May , Zimmermann Maike , Semrad Thomas , Wu Chun-Yi , Yu Aiming , Pan Chong-Xian , Henderson Paul T. , Kim Edward J.
Published: August 2019

Background

Combination 5-FU and oxaliplatin (FOLFOX) is standard therapy for colorectal cancer (CRC) in neoadjuvant, adjuvant, and metastatic settings. However, oxaliplatin neuropathy is the main dose-limiting toxicity that often necessitates dose reduction or discontinuation before progression. Prediction of oxaliplatin sensitivity in tumor cells would be valuable for dose optimization to maximize anti-tumorigenic effect and limit sensory neuropathy. Oxaliplatin functions via the formation of covalent drug-DNA adducts which ultimately inhibit cell replication and promote apoptosis. Detection of these drug-DNA adducts could serve as a biomarker of oxaliplatin efficacy. We therefore hypothesized that drug-DNA adduct levels induced by sub-therapeutic "diagnostic microdoses" are proportional to those induced by therapeutic doses and are predictive of tumor responses.

Methods

The feasibility of this predictive diagnostic microdosing approach was assessed in CRC cell culture and a pilot clinical trial of six CRC patients. Four CRC cell lines were cultured with therapeutically relevant (100 µM) or diagnostic microdose (1 µM) concentrations of [14C]oxaliplatin. The C-14 label enabled quantification of oxaliplatin-DNA adduct levels with accelerator mass spectrometry, an ultrasensitive isotope analysis technique. Adduct formation was correlated with oxaliplatin cytotoxicity as measured by the MTT viability assay. In a pilot clinical trial (NCT02569723), six CRC patients received an intravenous diagnostic microdose containing [14C]oxaliplatin prior to treatment and a second [14C]oxaliplatin dose during FOLFOX chemotherapy. Adduct levels from peripheral blood mononuclear cells (PBMC), used as surrogates for tumor cells, were correlated to mean tumor volume change of evaluable target lesions.

Results

Drug-DNA adduct levels were linearly proportional between microdoses and therapeutically relevant concentrations in cell culture (R2 = 0.91, p < 0.0001) and patient samples (R2 = 0.63, p < 0.0001). Plasma pharmacokinetics of microdoses were consistent with those of therapeutic dosing. No microdose-associated toxicity was observed in this patient population. PBMC adduct levels significantly correlated with tumor shrinkage (microdose: R2 = 0.82, p = 0.033; therapeutic dose: R2=0.65, p = 0.099).

Conclusion

Diagnostic microdosing with oxaliplatin is feasible and is predictive of oxaliplatin sensitivity in CRC patients.

Citation Format: Justin Chen, May Cho, Maike Zimmermann, Thomas Semrad, Chun-Yi Wu, Aiming Yu, Chong-Xian Pan, Paul T. Henderson, Edward J. Kim. Microdose induced oxaliplatin-DNA adducts as a predictive biomarker of response in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT055.
Abstract CT062: A Phase I study of the safety and immunogenicity of personalized mutation-derived tumor vaccine and treatment fields in patients with newly diagnosed glioblastoma
Authors: Hormigo Adilia , Rubinsteyn Alex , Kodysh Julia , Blazquez Ana , Bhardwaj Nina
Published: August 2019

Glioblastoma (GBM) like other cancers generates mutation-derived tumor antigens (MTA) arising from somatic mutations such as insertions, deletions, nucleotide substitutions and gene fusions. These mutations can be recognized by the immune system, providing a specific target for antitumor therapy. The use of TTFields, a modality that utilizes low intensity electric fields that disrupt cell division, increases progression-free survival and overall survival in newly diagnosed GBM. Among several mechanisms of action, TTFields potentially induces an immunogenic cell death. Therefore, we combined this modality treatment with the standard of care of adjuvant Temozolomide chemotherapy and a personalized vaccine for newly diagnosed GBM in a single-arm, open label clinical trial. Phase Ia tests the safety and immunogenicity of MTA-vaccine and phase Ib tests the safety and immunogenicity of the vaccine in conjunction with the TTFields. RNA and DNA are extracted from tumor specimens and blood, and whole-exome sequencing and mRNASeq is performed. HLA typing is obtained for each patient. MTA are identified using a computational pipeline for prediction and ranking according to both HLA binding affinity and expression. A maximum of 10 peptides is selected for the synthesis of each patient vaccine. Patients receive 14 doses of the MTA-vaccine in combination with 14 doses of poly-ICLC, the vaccine adjuvant. We have found mutation-derived tumor antigens in all patients included in phase Ia despite a low mutational load in GBM patients. The patients that took part in phase Ia have received multiple vaccinations spanning over 6 months of treatment. The vaccine has been well tolerated without serious adverse events. Quantitative and qualitative changes in epitope-specific circulating T-cell response is currently being evaluated.

Citation Format: Adilia Hormigo, Alex Rubinsteyn, Julia Kodysh, Ana Blazquez, Nina Bhardwaj. A Phase I study of the safety and immunogenicity of personalized mutation-derived tumor vaccine and treatment fields in patients with newly diagnosed glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT062.
Abstract CT078: Cyfi: Results from a Phase Ib expansion cohort of anti-hepatocyte growth factor and cytarabine in relapsed and refractory AML
Authors: Wang Victoria E. , Andreadis Charalambos
Published: August 2019

Objective: Patients with AML who are refractory to induction therapy or relapse within 1 year have poor outcomes. Elevated serum hepatocyte growth factor (HGF) level is an adverse prognostic factor. Pre-clinical models have shown that myeloid blasts produce HGF in an autocrine fashion and pharmacologic blockade of the HGF/c-Met axis sensitizes blasts to cell death. We initiated a Phase Ib study with dose expansion cohort study to assess the safety and tolerability of the anti-HGF antibody ficlatuzumab with cytarabine in AML patients who are refractory to 7+3 or have relapsed within 1 year of induction.

Methodology: The 3 x 3 design was used for the Phase I with an expansion cohort of an additional 13 patients accrued and treated at the MTD. Ficlatuzumab was administered in escalated dosing of 10, 15, or 20 mg/Kg for 4 doses every 2 weeks starting on day 1. 20 mg/Kg dose was used for the expansion cohort. Cytarabine was administered at a fIxed dose of 2 g/m2 on days 2-7. PBMCs were collected at defined time points. Differential expression from multiplexed single cell RNA sequencing (scRNAseq) was used to assess biomarkers predictive of response.

Results: Of the total 18 patients accrued thus far, 16 are evaluable. 5 had progressive disease, and 11 responded, all complete responses. Most frequent grade 3/4 TEAEs were febrile neutropenia, LFT abnormalities, and electrolyte disturbance. There was 1 death from sepsis and multi-organ failure on day 23, following ANC recovery, from the disease, and 1 patient who withdrew from the study due to grade 4 gastrointestinal bleed, likely ficlatuzumab related, both prior to response assessment. scRNA sequencing identified a TNF alpha and IFN gamma inflammatory signature that correlates with response to ficlatuzumab at count recovery.

Conclusion: Cytarabine and ficlatuzumab is a safe combination with promising efficacy in high risk relapsed and refractory AML. This combination is warranted in further Phase II studies. scRNAseq may be used to identify biomarkers of response.

Citation Format: Victoria E. Wang, Charalambos Andreadis. Cyfi: Results from a Phase Ib expansion cohort of anti-hepatocyte growth factor and cytarabine in relapsed and refractory AML [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT078.
Abstract CT084: Long-term exposure (LTE) to Tislelizumab, an investigational anti-PD-1 antibody, in a first-in-human Phase I study
Authors: Desai Jayesh , Markman Benjamin , Friedlander Michael , Gan Hui , Horvath Lisa , Townsend Amanda , Millward Michael , Jameson Michael , Yen Chia-Jui , Hou Ming-Mo , Hou Jeannie , Wu John , Liang Liang , Deva Sanjeev
Published: August 2019

Background Tislelizumab (BGB-A317), an investigational monoclonal antibody with high affinity and specificity for PD-1, was engineered to minimize binding to FcγR on macrophages in order to abrogate antibody-dependent phagocytosis, a potential mechanism of resistance to anti-PD-1 therapy. Previous reports from early phase studies suggest tislelizumab was generally well tolerated and had antitumor activity in patients (pts) with advanced solid tumors. Clinical effects of tislelizumab LTE (>12 mo) in pts enrolled in the first-in-human study (NCT02407990) are presented here.

Methods Patients with advanced solid tumors received IV tislelizumab 0.5, 2, 5, or 10 mg/kg Q2W, 2 or 5 mg/kg administered Q2W or Q3W, or 200 mg IV Q3W. Antitumor activity was assessed by RECIST v1.1 criteria; PD-L1 expression was retrospectively assessed with the VENTANA PD-L1 (SP263) assay.

Results As of 31 Aug 2018, 63 of the 451 pts received tislelizumab for >12 mo. In these 63 pts, median age was 64 yr and 70% had received ≥1 prior systemic therapy. Tislelizumab LTE was most common in NSCLC (n=9), HCC (n=7), and bladder and ovarian (n=5 each) cancers. Four of the 5 pts who achieved CR during this study had LTE to tislelizumab (Table); all 4 pts were PD-L1+ (≥1% expression on tumor cells). Across the LTE cohort, ORR was 66.7%; PR and SD were observed in both PD-L1+ and PD-L1- tumors. The median time to CR/PR (3.7 mo) and duration of CR/PR (21.1 mo) were longer in pts with LTE than pts who responded but did not remain on treatment for >12 mo (2.1 and 6.3 mo, respectively). Rash was the only treatment-related AE (TRAE) reported in ≥15% of pts. Most TRAEs were of mild or moderate severity; arthritis, diarrhea, fatigue, granuloma, hyperglycemia, and lichenoid keratosis (n=1 each) were the only grade ≥3 TRAEs reported with tislelizumab LTE.

Conclusion Tislelizumab remained well tolerated for >12 mo and elicited durable responses in pts with a variety of tumor types regardless of PD-L1 status.

PD-L1+ (n=35)PD-L1- (n=22)Missing (n=6)Total (N=63)CR4004 (6.3%)PR2113438 (60.3%)SD99220 (31.7%)PD1001 (1.6%)Abbreviations: CR, complete response; PD, progressive disease; PR, partial response; SD stable disease.

Citation Format: Jayesh Desai, Benjamin Markman, Michael Friedlander, Hui Gan, Lisa Horvath, Amanda Townsend, Michael Millward, Michael Jameson, Chia-Jui Yen, Ming-Mo Hou, Jeannie Hou, John Wu, Liang Liang, Sanjeev Deva. Long-term exposure (LTE) to Tislelizumab, an investigational anti-PD-1 antibody, in a first-in-human Phase I study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT084.
Abstract CT111: Preclinical justification for the Phase II clinical trial ADI-PEG 20 in combination with gemcitabine and docetaxel for the treatment of soft tissue sarcoma
Authors: Prudner Bethany C. , Rathore Richa , Godec Abigail , Chang Samuel F. , Hirbe Angela , Tine Brian A. Van
Published: August 2019

The response to acute and long-term arginine starvation results in an adaptive metabolic reprogramming that can be harnessed for therapeutic opportunities in ASS1-negative tumors. In examining modulations within the pyrimidine pathway, we identified that the addition of docetaxel (DTX) to cells treated with arginine deiminase (ADI-PEG20) results in a stabilized and translocation c-Myc to the nucleus. This effect drives increased hENT1 cell surface, allowing for the uptake of gemcitabine (GEM) and related analogues. This can be attenuated via a c-Myc/Max heterodimerization inhibitor. Combination experiments showed that the addition of ADI-PEG20 to the combination of GEM and DTX was synergistic. In addition, this combination can be used to overcome intrinsic hENT related GEM resistance to increase GEM uptake. In vivo studies indicate that the triple combination was optimal for tumor growth inhibition providing the preclinical mechanisms and justification for the ongoing clinical trial NCT03449901 ADI-PEG 20 in Combination With Gemcitabine and Docetaxel for the Treatment of Soft Tissue Sarcoma.

Citation Format: Bethany C. Prudner, Richa Rathore, Abigail Godec, Samuel F. Chang, Angela Hirbe, Brian A. Van Tine. Preclinical justification for the Phase II clinical trial ADI-PEG 20 in combination with gemcitabine and docetaxel for the treatment of soft tissue sarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT111.