bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract C117: Development of autologous tumor: TIL coculture from patient-derived samples for in vitro preclinical immuno-oncology studies
Authors: Hinata Kaede , Cuff Patrice , Liang Wei , Broudy Thomas 
Published: March 2020

Abstract
For over two decades, tumor infiltrating lymphocytes (TILs) have been utilized as an adoptive cell transfer therapy to treat cancer in clinical trials. In combination with added immunotherapy treatments, such as IL-2 or anti-CTLA4, TIL therapy has further improved clinic response rate. Following isolation from a patient’s own tumor specimen, TILs that recognize tumor-specific known or unknown antigens can be expanded. These cells are then infused back into tumor-bearing patients to attack the autologous cancer cell population. Despite progress in clinical research, preclinical and translational models of autologous tumor:TIL coculture have been difficult to establish. Our present study established autologous tumor:TIL coculture methods directly from patient tumor specimens. Models were established from BioDuro’s viably frozen tumor bank, containing more than 100,000 patient biospecimens. Each patient specimen contains a heterogeneous population of cell types, including tumor cells, TILs and TAFs (tumor associated fibroblasts). To establish our patient-derived coculture system, these cell populations were expanded under specific growth conditions to then serve as components of the coculture system. In this study, TILs and monocytes from a cohort of patient tumor specimens were analyzed by FACS to evaluate surface receptor expression. Leukocytes, T cells, monocytes and B cells were marked with CD45, CD3, CD14, CD20, respectively. PD-1 expression levels were also evaluated, with the results showing significant individual variation of expression on cell subpopulations. Established methods were used for the expansion of TILS, supporting either CD4 or CD8 T cell phenotypes. Autologous tumor cells were also expanded, concurrently. Afterward, TILs and tumor cells were cocultured and tumor cell killing was monitored using high content fluorescent image cytometry. We found that autologous patient-matched TILs showed preferential tumor cell killing as compared to coculture with patient-unmatched TILs. This response was observed in a dose-dependent manner. In conclusion, BioDuro’s biobank of viably frozen tumor and TIL patient specimens serves as an expansive and powerful immunooncology resource to establish patient-derived autologous tumor:TIL coculture assays. These assays, along with the ability to readily study larger populations of cancer patients, provide a powerful in vitro platform to test immunooncology drug candidates.
Citation Format: Kaede Hinata, Patrice Cuff, Wei Liang, Thomas Broudy. Development of autologous tumor: TIL coculture from patient-derived samples for in vitro preclinical immuno-oncology studies [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 C117. doi:10.1158/1535-7163.TARG-19-C117
Abstract C118: Transcriptome comparison of orthotopic and subcutaneous patient-derived xenografts
Authors: jiang xiaoqian , li henry , guo sheng 
Published: March 2020

Abstract
Background: Patient-derived xenograft (PDX) models are used for in vivo evaluation of oncology drugs. Tumors from cancer patients are implanted into immunodeficient mice either subcutaneously or orthotopically. Subcutaneous PDX models are more prevalently used because of easy operation and monitoring, albeit increasing evidence showed that the immune microenvironment in subcutaneous tissue is different from the original tract and is therefore less patient-like. By contrast, orthotopic PDX models have the capability to metastasize and match that of the donor patient well, but resulting in extensive inflammation. Previous studies showed that tumor location may affect immune response or result in different behavior of tumor expansion in mouse models [1-4]. There is yet insufficient study on the transcriptomic difference between subcutaneous and orthotopic PDX models. Method: Tumors for 10 PDX models—including four breast, four colorectal, and two liver cancers—were used to establish pairs of subcutaneous and orthotopic models. Tumors were dissected when reaching about 1000mm3, and subjected to transcriptomic profiling by RNAseq for gene expression comparison. Results: Using traditional alignment-based RNAseq analysis, we found hundreds of genes with different expression levels between subcutaneous and orthotopic models for the three cancers. Gene ontology analysis revealed that these differentially expressed genes are highly enriched in protein translation related biological processes and pathways, such as maturation of SSU-rRNA, cap-dependent translation initiation, and peptide chain elongation. Conclusion: Subcutaneous and orthotopic PDX models differ in transcriptomes that may affect protein translation and warrant further proteomics investigations. Reference: 1, Zhang, Yong, et al. “Real–Time GFP Intravital Imaging of the Differences in Cellular and Angiogenic Behavior of Subcutaneous and Orthotopic Nude–Mouse Models of Human PC–3 Prostate Cancer.” Journal of cellular biochemistry 117.11 (2016): 2546-2551. 2, Zhao, Xianda, et al. “Tumor location impacts immune response in mouse models of colon cancer.” Oncotarget 8.33 (2017): 54775. 3, Zhang, Yi, et al. “Establishment of a murine breast tumor model by subcutaneous or orthotopic implantation.” Oncology letters 15.5 (2018): 6233-6240. 4, Nakano, Kiyotaka, et al. “Difference in morphology and interactome profiles between orthotopic and subcutaneous gastric cancer xenograft models.” Journal of toxicologic pathology 31.4 (2018): 293-300.
Citation Format: xiaoqian jiang, henry li, sheng guo. Transcriptome comparison of orthotopic and subcutaneous patient-derived xenografts [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 C118. doi:10.1158/1535-7163.TARG-19-C118
Abstract C123: Loss of tumor suppressor NDRG2 promotes cell proliferation and migration/invasion of oral squamous cell carcinoma cells
Authors: Yoon Hye-Jung , Ahn Chi-Hyun , Kim Ji-Hoon 
Published: March 2020

Abstract
More than 90% of oral cancers are diagnosed as oral squamous cell carcinoma (OSCC). Although there has been much understanding of the molecular pathogenesis of OSCC, the majority of patients still die from local recurrence and metastasis of tumors. Recently, N-myc downstream-regulated gene 2 (NDRG2) has been known as a tumor suppressor gene in various types of malignancies, but its function and molecular mechanism in OSCC are very little known. To verify the functional roles of NDRG2 in OSCC, we screened six OSCC cell lines first using western blotting. Among the 6 cell lines, HN22 cell line showing high expression of NDRG2 was selected and treated with anti-NDRG2 small interfering RNA (siRNA). The effect of NDRG2 on cell proliferation was evaluated at 24h, 48h, and 72h after treatment of 50nM of anti-NDRG2 siRNA. In addition, in vitro migration and invasion assays were performed after same treatment. Western blot analysis showed variable levels of NDRG2 in 6 OSCC cell lines. Normal human oral keratinocyte as a positive control expressed very high level of NDRG2. Among 6 OSCC cell lines, three cell lines had very low level or absence of NDRG2, but remaining three cell lines including HN22 cell had moderate to high level of NDRG2. After 24h treatment of anti-NDRG2 siRNA on the HN22 cells, NDRG2 expression was significantly downregulated. In cell proliferation assay after treatment of 50nM of anti-NDRG2 siRNA, there was a significant increase in cell growth of HN22 cells when compared with the control (p=.005). In wound healing assay, NDRG2-downregulated HN22 cell line showed much more closure of wound area when compared with the control (p=.007). From the invasion assay after treatment, there was a significant increase in invasion ability of HN22 cells (p&lt;.001). These date indicate that loss of function of NDRG2 as a tumor suppressor can enhance the cell growth, migration, and invasion of OSCC cells. Our results suggest that loss of NDRG2 expression plays a role in tumor progression of OSCC by enhancing tumor cell growth, motility, and invasiveness. Therefore, recovering loss of function of NDRG2 may be a new therapeutic strategy for OSCC treatment.
Citation Format: Hye-Jung Yoon, Chi-Hyun Ahn, Ji-Hoon Kim. Loss of tumor suppressor NDRG2 promotes cell proliferation and migration/invasion of oral squamous cell carcinoma 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 C123. doi:10.1158/1535-7163.TARG-19-C123
Abstract C132: Fusobacterium nucleatum in colorectal carcinoma and expression of multiple immune checkpoint molecules on T cells
Authors: Kim Han Sang , Kim Wonkyu , Kim Chang Gon , Yoo Jinseon , Paik Soonmyung , Shin Eui-Cheol , Kim Tae-Min , Kim Hoguen , Ahn Joong Bae 
Published: March 2020

Abstract
Background: Fusobacterium nucleatum (Fn) appears to play a role in colorectal cancer carcinogenesis through suppression of antitumor immune response. This study aims to characterize immune-related signatures, including the expression of multiple immune checkpoint molecules in tumor-infiltrating T cells. Methods: We measured Fn DNA by quantitative PCR in a total of 155 colorectal cancer tissues. Gene expression profile was studied using microarray technology to determine the consensus molecular subtypes (CMS) and immune-related signatures using CIBERSORT and Gene Set Enrichment Analysis (GSEA), respectively. Expression of immune checkpoint molecules (PD-1, TIM-3, and TIGIT) on CD8+ cytotoxic T cells and the ratio of FOXP3-CD4-/FOXP3+CD4 were examined using multicolor flow cytometric analysis. Results: All patients had stage III disease underwent curative surgery. Fn infection was observed in 43% of patients and was associated with recurrence (Fisher’ exact test, P=0.002) and decreased overall survival (log-rank test, P&lt;0.001). The distribution of CMS subtypes was CMS1/2/3/4/mixed (14%/24%/15%/27%/20%), which is comparable to previous data. Fn infection was associated with the platinum-resistant signature, hypoxia pathway, and stem cell pathways in the GSEA. Interestingly, Patients with Fn infection showed a trend toward enrichment in CMS4, mesenchymal subtype with the inflamed immune phenotype (9% vs. 17% with Fn-infection, P=0.24). The number of cytotoxic CD8+ T cells showed a trend toward decreased expression in patients with Fn infection. Furthermore, higher expression of inhibitory immune checkpoints (PD-1, P&lt;0.01; TIM-3, P&lt;0.05; TIGIT, P&lt;0.05) was observed on CD8+ cytotoxic T cells of patients with Fn infection, compared to that of Fn negative cases. Besides, the regulatory FOXP3+CD4+ cells were increased in patients with Fn infection, supporting the suppressive role of Fn infection in the tumor immune microenvironment (P&lt;0.05). Conclusions: The adverse prognostic impact of Fn infection depends on the suppression of T cell-mediated immune response by increased expression of inhibitory immune checkpoints on cytotoxic T cell and by increased regulatory T cells.
Citation Format: Han Sang Kim, Wonkyu Kim, Chang Gon Kim, Jinseon Yoo, Soonmyung Paik, Eui-Cheol Shin, Tae-Min Kim, Hoguen Kim, Joong Bae Ahn. Fusobacterium nucleatum in colorectal carcinoma and expression of multiple immune checkpoint molecules on T 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 C132. doi:10.1158/1535-7163.TARG-19-C132
Abstract C133: Effective targeting of KRAS-driven tumors by the first-in-class, orally bioavailable SOS1::KRAS inhibitor BI-3406
Authors: Hofmann Marco H , Gmachl Michael , Ramharter Jürgen , Savarese Fabio , Sanderson Michael P , Gerlach Daniel , Trapani Francesca , Botesteanu Dana-Adriana , Kessler Dirk , Ettmayer Peter , Arnhof Heribert , Gerstberger Thomas , Schnitzer Renate , Kofink Christiane , Stadtmueller Heinz , Wunberg Tobias , O’Connell Jonathan C. , Mendes Rachel L , Richard David , Saldanha Adrian S , Moll Juergen , Pearson Mark , McConnell Darryl B , Kraut Norbert 
Published: March 2020

Abstract
KRAS is the most frequently mutated oncogene with high prevalence in pancreatic, colorectal, and non-small cell lung cancers. KRAS signaling is tightly regulated and release of negative feedback pathways have limited the clinical efficacy of inhibitors of the downstream MAPK signaling in the KRAS mutant context. Here we report the discovery of BI-3406, a highly potent and selective, first-in-class, orally bioavailable SOS1::KRAS inhibitor that binds to the catalytic domain of the guanine exchange factor SOS1 thereby preventing the interaction with KRAS-GDP. In KRAS-dependent cancers, BI 3406 potently reduces the formation of GTP-loaded KRAS, and inhibits MAPK pathway signaling. Down-modulation of this signaling cascade by BI-3406 in KRAS G12 or G13 mutant cells results in a cytostatic effect. In monotherapy, the observed pathway modulation translates into marked anti-tumor efficacy in KRAS mutant xenografts. BI-3406 blocks the negative feedback induced by MAPK inhibition and consequently sensitizes KRAS-dependent cancers to MEK inhibitor treatment resulting in complete blockade of the pathway, which translates into tumor regression in vivo. Collectively, our data suggest that SOS1 inhibition is a promising therapeutic strategy to treat KRAS dependent cancers in combination with MEK inhibition, with the aim to deliver meaningful survival benefits for cancer patients.
Citation Format: Marco H Hofmann, Michael Gmachl, Jürgen Ramharter, Fabio Savarese, Michael P Sanderson, Daniel Gerlach, Francesca Trapani, Dana-Adriana Botesteanu, Dirk Kessler, Peter Ettmayer, Heribert Arnhof, Thomas Gerstberger, Renate Schnitzer, Christiane Kofink, Heinz Stadtmueller, Tobias Wunberg, Jonathan C. O’Connell, Rachel L Mendes, David Richard, Adrian S Saldanha, Juergen Moll, Mark Pearson, Darryl B McConnell, Norbert Kraut. Effective targeting of KRAS-driven tumors by the first-in-class, orally bioavailable SOS1::KRAS inhibitor BI-3406 [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 C133. doi:10.1158/1535-7163.TARG-19-C133
Abstract CN01-02: Maximizing the potential of bispecific antibodies for cancer drug development
Authors: Parren Paul W H I 
Published: March 2020

Abstract
Landmark advances in the engineering and development of bispecific antibodies (bsAbs) are enabling unprecedented versatility in therapeutic antibody concepts. Currently more than 20 different commercialized technology platforms are available for bsAb creation and development, two bsAbs are marketed, of which one in cancer, and over 85 are in clinical development. A defining bsAb feature is their potential for novel functionalities — that is, activities that do not exist in mixtures of the parental or reference antibodies. These so-called obligate bsAb have high potential for the development of novel and differentiating cancer drugs. Bispecific T cell engagers (bs-TCE) are a prominent class of obligate bsAb for targeted cancer immunotherapy. Classical bs-TCE designs consist of a tumor antigen-binding domain combined with a binding domain against the CD3 T-cell receptor-signaling complex. Irrespective of the bispecific antibody format used, CD3-based bs-TCEs have a number of disadvantages, which in part are explained by the fact that they activate all T-cells irrespective of lineage, which associates with serious adverse events as a result of exaggerated T cell activation and cytokine release in some patients, and limited efficacy due to T suppressor cell activation in others. The development of bs-TCEs with increased tumor selectivity to widen the therapeutic window has high potential. The engagement of γδ T cells, and Vγ9Vδ2-T cells in particular, is in this context of particular interest. This γδ T cell subset has been shown to display powerful innate anti-tumor immune effector activity with an ability to infiltrate human tumors in which its abundance has been shown to positively correlate with patient survival. The potential of bsTCEs designed to engage Vγ9Vδ2-T cells as an approach for the development of novel cancer immunotherapies will be discussed.
Citation Format: Paul W H I Parren. Maximizing the potential of bispecific antibodies for cancer 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 CN01-02. doi:10.1158/1535-7163.TARG-19-CN01-02
Abstract CN07-02: Immunotoxins targeting GPC3 for liver cancer
Authors: Ho Mitchell , Fleming Bryan D , Greten Tim F , Pastan Ira 
Published: March 2020

Abstract
Glypican-3 (GPC3) is a cell surface glypican that serves as a co-receptor for Wnt. Our laboratory has generated two high affinity antibodies targeting GPC3, HN3 and YP7. The HN3 human nanobody recognizes the N-lobe of GPC3, whereas the YP7 antibody recognizes the C-lobe of GPC3. Mutating residue F41 on the N-lobe of GPC3 inhibits binding of Wnt and the HN3 nanobody and inhibits activation of β-catenin. Both antibodies are fused to a fragment of Pseudomonas exotoxin A (PE38) to create recombinant immunotoxins. Interestingly, the HN3-derived immunotoxin (HN3-PE38) has superior antitumor activity as compared with the YP7-derived immunotoxin (YP7-PE38). Intravenous administration of HN3-PE38 causes regression of hepatocellular carcinoma (HCC) tumor xenografts in mice. Our study establishes GPC3 as a promising target for immunotoxin-based liver cancer therapy and demonstrates immunotoxin-induced tumor regression via dual mechanisms: inactivation of Wnt signaling via the HN3 nanobody and inhibition of protein synthesis via the PE bacteria toxin. However, immunogenicity and a short serum half-life may limit the ability of immunotoxins to transition to the clinic. To address these issues, we have recently engineered HN3-based immunotoxins to contain various deimmunized PE toxins. These new immunotoxins include HN3-T20, which is modified to remove domain II of the PE toxin and six T-cell epitopes. All of our immunotoxins display high affinity to human GPC3, with HN3-T20 having a KD value of 7.4 nM. A real-time cell growth inhibition assay demonstrates that a single dose of HN3-T20 at 1.6 nM is capable of inhibiting nearly all cell proliferation during the 10-day experiment. To enhance HN3-T20’s serum retention, we further engineer the HN3-T20 by adding a streptococcal albumin binding domain (ABD) and a llama single-domain antibody fragment (ALB1) specific for serum albumin. For the detection of immunotoxin in mouse serum, we develop a highly sensitive ELISA and find that HN3-ABD-T20 has a 45-fold higher serum half-life than HN3-T20 (326 min vs 7.3 minutes); consequently, HN3-ABD-T20 shows the best serum retention and remains detectible even after a 24-hour period in mice. Furthermore, addition of an albumin binding domain results in HN3-ABD-T20 mediated tumor regression in mice at a low dose (1 mg/kg). These data show that the albumin binding deimmunized immunotoxins are high potency therapeutics ready to be evaluated in clinical trials for the treatment of liver cancer.
Citation Format: Mitchell Ho, Bryan D Fleming, Tim F Greten, Ira Pastan. Immunotoxins targeting GPC3 for liver cancer [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 CN07-02. doi:10.1158/1535-7163.TARG-19-CN07-02
Abstract CN08-02: BMP signaling in glioblastoma
Authors: Miyazono Kohei , Tanabe Ryo 
Published: March 2020

Abstract
Glioma-initiating cells (GICs) play central roles in the initiation, progression, recurrence, tumor heterogeneity and drug resistance of glioblastoma. Bone morphogenetic proteins (BMPs) regulate normal embryonic neural development and exert anti-tumor activities in glioblastoma. However, its mechanisms of action remain to be elucidated. Through RNA-seq analyses, we have identified several BMP-target genes in GICs, including SMAD6, Noggin (NOG), Distal-less homeobox 2 (DLX2), and Paired-related homeobox 1 (PRRX1). DLX2 promotes apoptosis and neural differentiation of GICs. We also show that the expression of PRRX1 is induced by BMP stimulation, and that PRRX1 induces decrease in the CD133-positive GIC population and inhibits in vivotumorigenic activity of GICs. Of the two spliced isoforms of PRRX1, the longer form of PRRX1, but not the shorter form, is involved in the induction of differentiation of GICs through epigenetic regulation of the CD133/PROM1gene. In addition, we show that BMP-4 represses the expression of a transmembrane protein, which is preferentially expressed in mesenchymal subtype of glioblastoma. Potential use of these proteins for diagnosis and treatment of glioblastoma will be discussed.
Citation Format: Kohei Miyazono, Ryo Tanabe. BMP signaling in glioblastoma [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 CN08-02. doi:10.1158/1535-7163.TARG-19-CN08-02