bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract CT103: Rationale for flat dosing of AZD9150 by population pharmacokinetic analysis
Authors: Tong Xiao , Mugundu Ganesh , Scott Martin , Cook Carl , Lyne Paul , Al-Huniti Nidal , Xu Hongmei
Published: August 2018

Objective: AZD9150 (ISIS 481464) is a 16-nucleotide oligonucleotide antisense molecule designed to reduce expression of human STAT3 protein by targeted down-regulation of STAT3 messenger RNA and is currently in clinical development in advanced solid tumors. The recommended Phase 2 dose (RP2D) for AZD9150 is 3 mg/kg, dosed based on ideal body weight (IBW) with three loading doses on day 1, 3 and 5 in week 1 followed by weekly dosing thereafter. The objective of the current analysis was to assess the effect of body size on AZD9150 pharmacokinetics (PK) to determine the appropriate dosing approach using population PK modeling.

Methods: PK and demographics data from 3 phase I/II studies (481464-CS1 [NCT01563302], D5660C00001 [NCT01839604], D5660C00004 [NCT02499328]) in total of 126 patients with diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma (HCC) and squamous-cell carcinoma in head and neck (SCCHN), receiving doses ranging from 1 to 4 mg/kg as monotherapy or in combination with durvalumab, were used in the population PK analysis. Both 2-compartment and 3-compartment structure PK models were tested in the analysis. Patient characteristics including age, ideal body weight (IBW), race and sex were explored as covariates on relevant AZD9150 PK parameters in an automated forward inclusion followed by backward elimination procedure through use of the stepwise covariate modelling. Once a final model was identified, simulations (n=1000, body weight distribution 30 to 100 kg) were conducted to compare AZD9150 exposures at the RP2D of 3 mg/kg versus a flat dose of 200 mg.

Results: A 2-compartment model with linear elimination well described the data. None of the covariates (inc. IBW) were found to significantly impact AZD9150 PK. The final PK model parameter estimate for clearance (CL), central volume of distribution (V1) and peripheral volume of distribution (V2) are 3.11 L/h, 5.56 L and 87.3 L, respectively. Model based simulations demonstrate that systemic exposure of AZD9150 was similar between 3 mg/kg versus 200 mg dose with slightly less overall between-patients variability with the flat dosing regimen.

Conclusion: Population PK modeling supports the potential switch of dosing approach from 3 mg/kg dose to a flat dose of 200 mg. The AZD9150 flat dosing rationale and schedule has been granted approval by FDA (for IND 137525) and also by MHRA (for BISCAY trial) as the first protocol / IMPD to be approved in the EU. Flat dose of 200 mg is currently being evaluated in ongoing clinical studies.

Citation Format: Xiao Tong, Ganesh Mugundu, Martin Scott, Carl Cook, Paul Lyne, Nidal Al-Huniti, Hongmei Xu. Rationale for flat dosing of AZD9150 by population pharmacokinetic analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT103.
Abstract LB-006: Intracellular protein therapy with SOCS3 utilizing aMTD-enabled macromolecule intracellular transduction technology (MITT)
Authors: Choi Youngsil , Shin Seulmee , Lee Kuysook , Jo Daewoong
Published: August 2018

Background Macromolecule intracellular transduction technology (MITT), a platform for the discovery/development of new medicinal drugs, is enabled with a new generation of hydrophobic cell-penetrating peptides (CPPs), termed advanced macromolecule transduction domains (aMTDs). Originally derived from the hydrophobic signal peptides of secreted proteins, earlier generations of MTDs were hampered by poor solubility and less efficient cell-permeability. The MITT platform addresses these limitations, first, by the design of synthetic aMTD sequences that incorporate 6 critical factors found to enhance the intracellular delivery of proteins into cells and tissues and second, by the use of humanized solubilization domains to enhance the solubility of aMTD-containing recombinant proteins. Object The goal of this study is to develop and evaluate an aMTD-SOCS3 (suppressor of cytokine signaling 3) protein (iCP-SOCS3) as cell-permeable biotherapeutic to selectively target tumors dependent on JAK/STAT signaling and to compare the cell-/tissue-penetration efficiencies of iCP-SOCS3 with a previous generation of cell-permeable SOCS3 (MTM-SOCS3: CP-SOCS3) enabled by a hydrophobic CPP, termed membrane translocating motif (MTM) derived from the signal peptide of fibroblast growth factor 4 (FGF4). Results Solubility (up to 10 mg/ml) and final yields of E. coli produced iCP-SOCS3 (60 mg/L) were sufficient for clinical development; whereas, CP-SOCS3 suffered from low solubility and yields (0.4 mg/ml and 0.2 mg/L, respectively). Cellular uptake of fluorescent iCP-SOCS3 was enhanced by 25-fold as compared to CP-SOCS3 as assessed by flow cytometry, and delivery and persistence of the protein in a variety of tissues (heart, lung, spleen, kidney, liver and brain) were similarly increased. In addition, aMTD promoted the cell-to-cell transfer of SOCS3 protein, and the transferred iCP-SOCS3 was biochemically active as assessed by inhibition of IFN-γ-induced phosphorylation of STAT1 and STAT3 with concomitant suppression of cancer-associated phenotypes in cultured cells and in mice bearing human tumor xenografts (90% inhibition of proliferation and 84% inhibition of migration in PANC-1 cells, and 85% suppression of pancreatic tumor growth, respectively). Conclusion These results provide further validation of MITT as a platform to develop protein-based biotherapeutics, and suggest the use of iCP-SOCS3 as a novel protein therapy against tumors dependent on JAK/STAT signaling.

Citation Format: Youngsil Choi, Seulmee Shin, Kuysook Lee, Daewoong Jo. Intracellular protein therapy with SOCS3 utilizing aMTD-enabled macromolecule intracellular transduction technology (MITT) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-006.
Abstract LB-007: Intracellular delivery of SOCS3 suppresses pancreatic cancer progression
Authors: Lee Kuysook , Shin Seulmee , Hwang Yunseo , Choi Youngsil , Jo Daewoong
Published: August 2018

Background JAK/STAT signaling, regulated by SOCS family proteins, plays a role in the progression of the inflammation, growth and metastasis of pancreatic adenocarcinoma. SOCS3 suppresses JAK kinase activity and degrades the activated cytokine receptor complex, and its loss contributes to pancreatic cancer development. We have developed advanced macromolecule transduction domains (aMTDs) to deliver recombinant proteins into to cells and tissues. In principle, aMTD enabled SOCS3 could provide an effective treatment for pancreatic tumors that depends on JAK/STAT signaling for growth or survival. Objective The present study utilized aMTD sequences to develop improved cell-permeable (iCP-) SOCS3 as a protein-based biotherapeutic against pancreatic cancer. Method Improved cell-permeable SOCS3 proteins, with and without a 12-amino acid aMTD sequence, were expressed at high yields in E. coli. The proteins also contained a solubilization domain (SD) to enhance the solubility of the recombinant proteins in physiological buffers. Results The iCP-SOCS3 exhibited high solubility, cell-/tissue-permeability and biological activity in pancreatic cancer cell lines (BxPC3, HPAC and PANC-1) with decreased endogenous SOCS3 levels, hyperactived JAK/STAT signaling and hypermethylated socs3 promoter region. The iCP-SOCS3 suppressed cancer-associated phenotypes (i.e., up to 90% inhibition of cell viability, 52% induction of apoptosis, 14.8% inhibition of cell cycle progression, 54% inhibition of migration and 61.5% inhibition of invasion) in pancreatic cancer cells dependent on elevated JAK/STAT signaling, whereas, these effects were not observed in non-cancer cells (H6c7, pancreatic duct epithelial cell) or with proteins lacking the aMTD sequence. In contrast, gemcitabine, a conventional pancreatic cancer therapeutic, showed undesirable cytotoxic effects including decreased cell viability and induced apoptosis in non-cancer cells. Furthermore, iCP-SOCS3 (30 mg/kg) induced tumor regression (-168% at day 18) in PANC-1 CDX model. In BxPC-3 orthotopic CDX model, iCP-SOCS3 suppressed tumor growth by 98% which was significantly higher compared to the suppressed tumor growth induced by gemcitabine (77%). Correlating to the tumor suppression, iCP-SOCS3 significantly decreased the secretion of TNF-α and the expression of p-STAT1, p-STAT3 and VEGF in tumor tissues. Conclusion These results provide further evidence that SOCS3 can function as a tumor suppressor and intracellular delivery of SOCS3 with iCP-SOCS3 may provide a novel protein therapy against pancreatic cancer.

Citation Format: Kuysook Lee, Seulmee Shin, Yunseo Hwang, Youngsil Choi, Daewoong Jo. Intracellular delivery of SOCS3 suppresses pancreatic cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-007.
Abstract LB-009: Intracellular delivery with SOCS3 suppresses angiogenesis in tumors
Authors: Jung Yukdong , Lee Kuysook , Choi Youngsil , Jo Daewoong
Published: August 2018

Introduction: Suppressor of cytokine signaling 3 (SOCS3) is a negative feedback regulator in the Janus kinase/signal transducer and activator of transduction (JAK/STAT) signaling pathway, in which it suppresses inflammation and cancer. The JAK/STAT pathway is activated in some tumors thus enhancing the expression of genes involved in proliferation and angiogenesis, notably, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiopoietin-1 and phospoinositide 3-kinase (PI3K)-protein kinase B (AKT).

Objective: The aim of this study is to develop an improved cell-permeable (iCP)-SOCS3 and determine its applicability as an anti-angiogenic agent to treat cancer.

Methods: The iCP-SOCS3 was expressed in E. coli as a 37 kDa recombinant protein fused to an advanced macromolecule transduction domain (aMTD) and humanized solubilization domain (SD). Uptake of a fluorescently tagged iCP-SOCS3 by cells and tissues was monitored by flow cytometry and confocal microscopy. Biological and biochemical activities of iCP-SOCS3 were determined by in vitro angiogenesis assays and changes in biomarker expression and anti-tumor activities were analyzed in mice bearing human tumor xenografts.

Results: The iCP-SOCS3 improved on an earlier generation of CP-SOCS3 fused to FGF4-derived membrane translocating motif (MTM) with regard to solubility, yield, delivery into cells and tissues and biological activity. The iCP-SOCS3 suppressed proliferation of human umbilical vein endothelial cells (HUVECs) by 60% and migration by 54% via inhibition of AKT/mTOR signaling pathway. Moreover, iCP-SOCS3 inhibited invasion of EC cells in PANC-1 and U87MG conditioned medium by 38% and 95%, respectively. In tube formation assay, iCP-SOCS3 reduced the number of branch points by 79% and tube length by 60% compared to the vehicle. In chick chorioallantoic membrane (CAM) assay, iCP-SOCS3 significantly inhibited angiogenesis by 83% in dose dependent manner compared to the reference drug, retinoic acid. The anti-angiogenic activity of iCP-SOCS3 was also shown in matrigel plug assay. The new vessel formation induced by U87-MG glioblastoma cells was significantly reduced by the treatment of iCP-SOCS3. In PANC-1, U87-MG and HepG2 cell-derived xenograft models, iCP-SOCS3 suppressed expression of pro-angiogenic factors including VEGF, ERK and angiopoietin 1.

Conclusion: These results validate iCP-SOCS3 targets in promoting angiogenesis and suggest ways to reduce angiogenesis in various types of tumors.

Citation Format: Yukdong Jung, Kuysook Lee, Youngsil Choi, Daewoong Jo. Intracellular delivery with SOCS3 suppresses angiogenesis in tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-009.
Abstract LB-036: Targeting the WNT-pathway in 3D-cultures of pancreatic cancer biopsies
Authors: Herpers Bram , Basten Sander , Yan Kuan , Dijkmans Thomas , Schueler Julia , Giesemann Torsten , Price Leo S.
Published: August 2018

Creating patient-derived xenograft (PDX) models from human cancers allows bio-banking, propagation, characterization and drug testing on a large panel of tumor variants. We developed 3D in vitro culture methods for PDX model-derived cell isolates. Each individual model was subjected to a wide panel of culture medium formulations, amongst which some media contain WNT-pathway stimulating components. We identified an absolute dependency of a pancreatic in vitro PDX tumor to the WNT-signaling potentiating factor R-Spondin, while other pancreas tumors showed little or no dependency. To further study the dependency of pancreatic PDX tumors on the functionality and signaling of the WNT-pathway, we exposed in vitro PDX cultures to increasing doses of anti-FZD7, anti-FZD8 and to the Porcupine inhibitor LGK974, which blocks WNT palmitoylation and therefore secretion. We also applied the same approach to establish cultures from patient-derived pancreatic cancer biopsies. Our findings are that the WNT-pathway is functional and contributing to pancreas cancer outgrowth and morphological differentiation to different extents in different patient-models. Our in vitro culture system in combination with 3D phenotypic analysis allows in-detail study of the WNT-pathway, as well as by supplementation or omission of secreted factors, addition of small molecule inhibitors and by exposure to antibodies. These in vitro models can be used to test novel compounds and antibodies targeting the WNT pathway and underscore the WNT pathway as a potential target in pancreatic cancer.

Citation Format: Bram Herpers, Sander Basten, Kuan Yan, Thomas Dijkmans, Julia Schueler, Torsten Giesemann, Leo S. Price. Targeting the WNT-pathway in 3D-cultures of pancreatic cancer biopsies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-036.
Abstract LB-051: An Astragalus-based Chinese herbal medicine extraction inhibits cancer stem cell growth and sensitizes of drug-resistant human non-small cell lung cancer cells for targeted therapy
Authors: Huang Tse-Hung , Hsu Hsuan-Min , Huang Chi-Ying F.
Published: August 2018

Chinese herbal medicine has been used extensively as a complementary intervention for many diseases including cancer, based on its relatively lower cytotoxicity and side-effects. The major criticism of its clinical use remains the scientific based evidence. Here, we described a precision-based translational approach to identify and validate an Astragalus-based herbal medicine (YQ1) as an antagonist of non-small cell lung cancer. Using various big data analysis approaches, we found that the L1000 profiles of YQ1was associated with anti-cancer stem cell signatures. It suggested that treatment of YQ1 might reduce cancer stem cell population. Experimentally, YQ1 treatment resulted in the significant reduction in lung cancer stem cell properties, including side population, sphere formation, stemness marker expression, and drug resistance. YQ1 may have anti-cancer stem cell ability via suppressing Wnt/β-catenin signaling to inhibit the self-renewal or/and proliferation of lung cancer stem cells. Notably, the combination of YQ1 and gefitinib significantly suppresses tumorigenesis in vivo, reflected by the down-regulation of EGFR signaling and β-catenin. Collectively, we provided preclinical evidence for using YQ1 as a cancer stem cell inhibitor to potentiate anti-tumor effects when combined with tyrosine-kinase inhibitors to significantly improve therapeutic efficacy in NSCLC patients.

Citation Format: Tse-Hung Huang, Hsuan-Min Hsu, Chi-Ying F. Huang. An Astragalus-based Chinese herbal medicine extraction inhibits cancer stem cell growth and sensitizes of drug-resistant human non-small cell lung cancer cells for targeted therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-051.
Abstract LB-055: Discoidin domain receptor 1 (DDR1) ablation promotes tissue fibrosis and hypoxia to induce aggressive, basal-like breast cancer
Authors: Drain Allison P. , Takai Ken , Lawson Devon A. , Littlepage Laurie E. , Karpuj Marcela , Kessenbrock Kai , Le Annie , Inoue Kenichi , Weaver Valerie M. , Werb Zena
Published: August 2018

Breast cancer studies implicate the discoidin domain receptor 1 (DDR1) in both pro- and anti-tumor roles. DDR1 is often overexpressed in breast tumors and its activation by collagen promotes tumor cell invasion and potentiates metastatic colonization by cell lines. However, DDR1 transfection into low DDR1 expressing tumor cells hinders cell migration. Moreover, low expression of DDR1 correlates with higher grade human tumors and associates with worse prognosis. To better understand the role of DDR1 in breast tumor progression and metastasis, we established a DDR1 knockout and crossed it into the MMTV-PyMT mouse. DDR1 loss compromised luminal cell adhesion and expanded the basal/myoepithelial population in adult mice. PyMT DDR1-/- tumors grew faster and had increased lung metastasis both in size and number of metastatic colonies. The DDR1-/- tumors broadly exhibited a more basal-like phenotype with enhanced fibrosis and were more hypoxic. Furthermore, these tumors contained more CD90+ CD24+ tumor cells and more K8+ K14+ cells surrounding necrotic regions. Taken together these data suggest DDR1 loss confers a growth advantage favoring basal cell expansion via impaired luminal adhesion and enhanced basal differentiation leading to more aggressive, basal-like disease.

Citation Format: Allison P. Drain, Ken Takai, Devon A. Lawson, Laurie E. Littlepage, Marcela Karpuj, Kai Kessenbrock, Annie Le, Kenichi Inoue, Valerie M. Weaver, Zena Werb. Discoidin domain receptor 1 (DDR1) ablation promotes tissue fibrosis and hypoxia to induce aggressive, basal-like breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-055.
Abstract LB-084: Dasatinib reduces tumor growth in xenograft models derived from human lung tumors with YES1 overexpression
Authors: Garmendia Irati , Bértolo Cristina , Ferrer Irene , Pajares María J , Ajona Daniel , Paz-Ares Luis , Pio Ruben , Montuenga Luis M , Agorreta Jackeline
Published: August 2018

Introduction Lung cancer accounts for 13.2% of all new cancer cases and is the leading cause of cancer-related death worldwide. For a reduction of mortality rates, new personalized therapies are needed. The great challenge is to identify the alterations responsible for the tumor malignant phenotype which should be targeted by specific therapeutic strategies. The aim of our study is to evaluate the use of YES1 kinase amplification and overexpression as a companion biomarker to predict sensitivity to dasatinib using patient-derived xenograft models. Previous results Our previous experiments showed that downregulation of YES1 expression by specific siRNAs impairs cell proliferation and survival in lung cancer cell lines with amplification and high expression of YES1 (YES1High). Inhibition of YES1 in cell lines with normal copy number and low expression (YES1Low) had no effect on cell proliferation. Moreover, YES1 downregulation induced apoptosis and inhibited invasion only in YES1High cells, and YES1 overexpression promoted proliferation in lung cancer cell lines.

Methods Eight patient-derived xenograft mouse models were used: two YES1High adenocarcinomas (ADC), two YES1Low ADC, two YES1High squamous cell carcinomas (SCC) and two YES1Low SCC. Small fragments of xenografted tumors (2-3 mm in diameter) were subcutaneously implanted into the right flank of 7-8-week-old BALB/c-Rag2−/−-IL2γc−/− immunodeficient mice. Mice were treated with dasatinib (60 mg/kg) or citric acid daily by oral gavage. Tumor volume was measured twice a week. Tumor volumes were calculated by the formula (LxW2)/2.

Results In those tumors harboring YES1 amplification (YES1High), dasatinib significantly reduced tumor growth. This treatment induced tumor response in both ADC and SCC PDX models. On the contrary, in tumors with low expression of YES1, dasatinib did not affect tumor growth in ADC or SCC PDX models.

Conclusions Dasatinib treatment significantly reduces tumor growth in YES1High PDX models but not in YES1Low models, providing support to our hypothesis that YES1 expression may be a useful companion biomarker for dasatinib as NSCLC treatment.

Citation Format: Irati Garmendia, Cristina Bértolo, Irene Ferrer, María J Pajares, Daniel Ajona, Luis Paz-Ares, Ruben Pio, Luis M Montuenga, Jackeline Agorreta. Dasatinib reduces tumor growth in xenograft models derived from human lung tumors with YES1 overexpression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-084.
Abstract LB-096: A gain-of-function screen identifies drivers of drug resistance in BRAFV600E melanoma cell lines
Authors: Feddersen Charlotte R. , Schillo Jacob L. , Vaughn Hayley R. , Voigt Andrew P. , Zhu Eliot Y. , Wadsworth Lexy S. , Stipp Christopher S. , Dupuy Adam J.
Published: August 2018

Melanoma is the deadliest form of skin cancer, and until recently patients had few treatment options upon diagnosis. Fortunately, this lack of treatment options changed with the discovery of vemurafenib, a highly selective kinase inhibitor that specifically targets the BRAFV600E mutant protein, found in ~50% of all melanoma cases. Initially, the kinase inhibitor provided complete or partial response to over 50% of patients and prolonged progression-free survival. However, the majority of patients relapse once tumors acquire resistance to vemurafenib. Some of the genetic resistance mechanisms are known, including BRAF amplification, expression of truncated BRAF, and RAS mutation. However, a significant portion of vemurafenib resistance cases is unexplained. While drug resistance can be delayed by combining vemurafenib with cobimetinib, a MEK inhibitor, mechanisms of resistance to this drug combination are not well characterized.

We performed a gain-of-function screen to identify drivers of drug resistance to vemurafenib or vemurafenib combined with cobimetinib. Briefly, A375 melanoma cells were engineered to express the hyperactive transposase SB100. These cells were then subsequently transfected with the mutagenic T2Onc3 transposon to produce a population of mutagenized A375 cells. Three weeks of vemurafenib treatment selected for cells with the acquired ability to grow in the presence of drug. Candidate driver mutations were then identified by profiling the sites of transposon insertion in drug-resistant cells to find genes that were recurrently over-expressed by transposon insertions. This led to the discovery of five genes that act as drivers of vemurafenib resistance when overexpressed in the human melanoma cell line A375. Candidate genes were then validated in multiple melanoma cell lines by assessing cell growth and viability in varying concentrations of vemurafenib.

An analogous screen was performed in A375 to identify drivers of resistance to the combination of vemurafenib and cobimetinib. In addition, we are assessing the diversity of drug resistance mechanisms by performing mutagenesis in three additional human melanoma cell lines.

The characterization of the novel drug resistance pathways identified here will improve treatment outcomes for patients with BRAFV600E mutant melanoma.

Citation Format: Charlotte R. Feddersen, Jacob L. Schillo, Hayley R. Vaughn, Andrew P. Voigt, Eliot Y. Zhu, Lexy S. Wadsworth, Christopher S. Stipp, Adam J. Dupuy. A gain-of-function screen identifies drivers of drug resistance in BRAFV600E melanoma cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-096.
Abstract LB-104: Targeting solid malignancies with TEGs: αβT cells

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ngineered to express a defined γδTCR in xenograft mice model
Authors: Johanna Inez , Straetemans Trudy , Heijhuurs Sabine , Jansen Koen , Rheenen Jacco van , Drost Jarno , Sebestyen Zsolt , Kuball Jurgen
Published: August 2018

γδT cells mediate cancer immune surveillance by sensing metabolic changes of malignant cells via their γδT cell receptor (TCR). Activation of γδTCR is independent of MHC molecules, making them a valuable addition to current treatment strategies. Moreover, γδTCR are able to differentiate between healthy and leukemic stem cells. This concept led to the development of next generation CAR T cells, so-called TEGs: αβT cells Engineered to express a defined γδTCR. A particular γ9δ2TCR, isolated from “clone 5”, has been selected as the candidate for clinical testing (TEG001). A purification strategy was introduced that combined maximal interference of γδTCR chains with endogenous TCR chains with a GMP-grade anti-αβTCR-bead-based depletion strategy to obtain a pure population of engineered immune cells (Straetemans et al. 2015). TEG001 cells showed a strong and broad recognition of hematological malignancies against both cell lines and primary AML (Marcu-Malina et al. 2011, Gründer et al. 2012). To further broaden the clinical implementation of this novel strategy we here investigated the in vivo efficacy profile against solid malignancies.

In this study, we evaluated clinical potency of TEG001 against human organoids engineered with most frequent mutations for colorectal cancer, including APC knockout (APCKO), constitutively active KRAS (KRASG12D), P53 knockout (P53KO), and SMAD4 knockout (SMAD4KO), also known as quadruple mutant organoids. These organoids are tumorigenic and can be successfully engrafted as solid tumor mass in immunodeficient mice (Fumagalli et al., 2017; Drost et al., 2015). Irradiated NSG mice received luciferase-positive quadruple mutant organoids subcutaneously and were subsequently infused with TEGs. Tumor growth was measured by bioluminescence imaging. TEG001 showed reduced tumor growth of quadruple mutant organoids compared to untreated and mock controls. Moreover, TEG001-treated mice showed increased overall survival relative to the control mice, followed up to 130 days after treatment.

Our current data shows that TEG001 mediates antitumor reactivity against quadruple mutant organoids as solid tumor model in vivo. This xenograft model allows further preclinical testing of next generation TEGs targeted to solid malignancies. Importantly, TEGs are a promising addition to the currently available immune therapeutic strategies as they target cancer as a metabolic disorder.

Citation Format: Inez Johanna, Trudy Straetemans, Sabine Heijhuurs, Koen Jansen, Jacco van Rheenen, Jarno Drost, Zsolt Sebestyen, Jurgen Kuball. Targeting solid malignancies with TEGs: αβT cells engineered to express a defined γδTCR in xenograft mice model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-104.
Abstract LB-120: Systematic association analysis of immune checkpoint blockade of PD-1 by RNA-seq data in advanced melanoma patients
Authors: Wang Shu
Published: August 2018

Immune checkpoint blockade with anti-programmed cell death protein-1 (PD-1) monoclonal antibodies (mAbs) provides long-term clinical benefits to 30-40% patients with advanced melanoma. In spite of this success, the majority of the melanoma patients (MPs) fail to respond to therapy. To further explore the mechanisms about how immunotherapy influences immune systems and cancers, we have developed a novel clinical trial with anti-PD-1 antibody (Pembrolizumab) and Peginterferon. In this trial, 43 melanoma patients were treated and had tumor biopsies pre- and post-treatment (week 12) that were used for transcriptomic studies using RNA-seq. Here, we report the data obtained from 21 MPs with lymph node metastases, including 10 responders and 10 non-responders. We found that lymph nodes and metastatic tumors show different gene expression pattern for response: besides immune signature which is expressed in lymph nodes also, metastatic tumors presents gene signature related to cell-ECM communication, vasculogenesis, etc. In addition, we found several pathways that favor response, such as IGF1/mTOR pathway. Our work gives a deeper insight into how anti-PD1 immunotherapy functions and how resistance against it occurs, which may predict clinical outcome in MPs treated with anti-PD-1 mAbs and Peginterferon.

Citation Format: Shu Wang. Systematic association analysis of immune checkpoint blockade of PD-1 by RNA-seq data in advanced melanoma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-120.
Abstract LB-132: MOSPD2: A novel target for Bi-specific Ab mediated killing of tumor cells
Authors: Mendel Itzhak , Salem Yaniv , Yacov Niva , Prhopheta-Meiran Osharat , Kafri Pinhas , Breitbart Eyal
Published: August 2018

Introduction: We have previously described motile sperm domain-containing protein 2 (MOSPD2) as essential for breast cancer metastasis in vitro and in vivo. In this study, we detected high expression of MOSPD2 in different organ tumors and cancer cells, and subsequently developed CD3xMOSPD2 BiTE, a firstly generated bi-specific antibody that directs killing of MOSPD2 expressing cancer cells.

Experimental procedure: The prevalence of MOSPD2 was evaluated by IHC in tissue microarrays layered with cores, representing multiple organ tumors and normal tissue. MOSPD2 abundance was scored according to the staining intensity on a scale from 0 to 3. Human monoclonal antibody against the extracellular region of human MOSPD2 was generated. To determine the presence of cell surface MOSPD2 expression, FACS analysis was performed on cell lines derived from different tumor origins. BiTE was generated to comprise the scFv region of anti-human CD3 and the scFv region of a human anti-human MOSPD2. Cancer cell lines were co-cultured with freshly isolated T-cells or effector CD8 T-cells in the presence of CD3xMOSPD2 or a control BiTE. Cancer cell survival rate and T cell activation was determined after 2-3 days by FACS.

Results: MOSPD2 expression was evident in the majority of organ tumors, with high prevalence in tumors such as colon, liver and breast. Normal tissue was either devoid of MOSPD2, or had low expression relative to its parallel tumor tissue. Within normal or normal adjacent tissue (NAT), staining could be detected primarily in lymphoid tissues where myeloid cells abundantly reside. FACS staining using a human anti-MOSPD2 mAb demonstrated that MOSPD2 is expressed on the surface of different cancer cell lines. Employing CD3xMOSPD2 BiTE on myeloid or solid tumor-derived cell lines induced T-cell activation and yielded profound killing of cancer cells.

Conclusions: IHC staining demonstrates MOSPD2 expression profile supportive of targeting cancer cells. CD3xMOSPD2 BiTE directs T-cells to MOSPD2 expressing cancer cells and induces their destruction in-vitro. Therefore, we propose CD3xMOSPD2 bi-specific antibodies as a potential therapy for the treatment of cancer via a unique target, directed for the destruction of hematological cancers and solid tumors expressing MOSPD2.

Citation Format: Itzhak Mendel, Yaniv Salem, Niva Yacov, Osharat Prhopheta-Meiran, Pinhas Kafri, Eyal Breitbart. MOSPD2: A novel target for Bi-specific Ab mediated killing of tumor cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-132.
Abstract LB-140: CAR T cells that target full-length MUC1 enrich for MUC1* and induce metastasis
Authors: Bamdad Cynthia C. , Stewart Andrew K. , Deary Luke T. , Smagghe Benoit J. , Huang Pengyu , Glennie Nelson D. , Swanson Tyler E.
Published: August 2018

Purpose: Investigate the rationale for CAR Ts that target full-length MUC1 (MUC1-FL) rather than the cleaved, growth factor receptor form, MUC1*. Over 75% of solid tumors aberrantly express MUC1. However, attempts to create an effective therapeutic by targeting MUC1-FL have all failed. No therapeutic that targets the cleavage product, MUC1*, has ever been tested in humans. On cancer cells, most of the MUC1-FL is enzymatically cleaved, releasing the bulk of the extra cellular domain and leaving a truncated transmembrane protein - MUC1*. There are currently two CAR T therapeutics (based on antibody 5E5 or SM3) being developed that target aberrant O-linked glycans. Importantly, MUC1* has no sites for O-linked glycans and is not recognized by 5E5 or SM3.

Experimental Procedures: Human ovarian cancer cells that express both MUC1-FL and MUC1* were implanted into the intraperitoneal (i.p.) space of female NSG mice. IVIS measurement verified engraftment. A cleavage enzyme that is overexpressed in most solid tumors and known to cleave MUC1-FL to MUC1* was injected into i.p. space to simulate the effect of a therapeutic that would eliminate cells expressing MUC1-FL. Human cancerous tissue specimens were probed with antibody VU4H5 that recognizes tandem repeats of MUC1-FL, 5E5 that recognizes aberrant O-linked glycans on tandem repeats of MUC1-FL, and anti-MUC1* antibody huMNC2.

Summary New Data: Elimination of cells expressing MUC1-FL, by cleaving MUC1-FL to MUC1*, caused tumors to grow dramatically (average 221-times in only 9 days) and metastasize. IHC studies showed that 30% of breast cancers do not express MUC1-FL but only express MUC1*. No tumor specimens were positive for MUC1-FL but negative for MUC1*. Our studies show that as tumor stage progresses, expression of MUC1* increases as MUC1-FL decreases. Over 95% of breast, 83% ovarian, 78% pancreatic and 71% lung cancers reacted with our anti-MUC1* antibody huMNC2-scFv (n>3,000). huMNC2-CAR44 T cells inhibited growth of MUC1* positive and MUC1-FL/MUC1* positive tumors, even after metastasis.

Conclusions: CAR T cells that target full-length MUC1 will enrich the tumor for MUC1*, increasing tumor growth and inducing metastasis.

Citation Format: Cynthia C. Bamdad, Andrew K. Stewart, Luke T. Deary, Benoit J. Smagghe, Pengyu Huang, Nelson D. Glennie, Tyler E. Swanson. CAR T cells that target full-length MUC1 enrich for MUC1* and induce metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-140.
Abstract LB-158: Longitudinal associations of lifetime adiposity with leukocyte telomere length and mitochondrial DNA copy number
Authors: Hang Dong , Nan Hongmei , Kværner Ane Sørlie , Vivo Immaculata De , Chan Andrew T. , Hu Zhibin , Shen Hongbing , Giovannucci Edward , Song Mingyang
Published: August 2018

Adiposity may cause adverse health outcomes by increasing oxidative stress and systemic inflammation, which can be reflected by altered telomere length (TL) and mitochondrial DNA copy number (mtCN) in peripheral blood leukocytes. However, little is known about the influence of lifetime adiposity on TL and mtCN in later life. This study was performed to investigate the associations of lifetime adiposity with leukocyte TL and mtCN in 9613 participants from the Nurses' Health Study. A group-based trajectory modelling approach was used to create trajectories of body shape from age 5 through 60 years, and a genetic risk score (GRS) was created based on 97 known adiposity susceptibility loci. Associations of body shape trajectories and GRS with dichotomized TL and mtCN were assessed by logistic regression models. After adjustment for lifestyle and dietary factors, compared with the lean-stable group, the lean-marked increase group had higher odds of having below-median TL (OR = 1.18, 95% CI: 1.04, 1.35; P = 0.01), and the medium-marked increase group had higher odds of having below-median mtCN (OR = 1.28, 95% CI:1.00,1.64; P = 0.047). There was a suggestive trend toward lower mtCN across the GRS quartiles (P for trend = 0.07). In conclusion, telomere attrition may be accelerated by marked weight gain in middle life, whereas mtCN is likely to be reduced persistently by adiposity over the life course. The findings indicate the importance of lifetime weight management to preserve functional telomeres and mitochondria.

Citation Format: Dong Hang, Hongmei Nan, Ane Sørlie Kværner, Immaculata De Vivo, Andrew T. Chan, Zhibin Hu, Hongbing Shen, Edward Giovannucci, Mingyang Song. Longitudinal associations of lifetime adiposity with leukocyte telomere length and mitochondrial DNA copy number [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-158.
Abstract LB-159: A multi-omic study reveals BTG2 as a reliable prognostic marker for early-stage non-small cell lung cancer
Authors: Shen Sipeng , Su Li , Zhang Ruyang , Loehrer Elizabeth , Lanuti Michael , Diao Nancy , Christiani David
Published: August 2018

Background B-cell translocation gene 2 (BTG2), which functions as a tumor suppress gene, has been reported to be involved in several cancers. However, no study has focused on its role in lung cancer progression or prognosis. We aimed to investigate the role of BTG2 in early-stage non-small cell lung cancer (NSCLC) survival.

Patients and methods This study included 1,230 early-stage (1-2) surgically treated NSCLC patients with methylation and expression data from five international cohorts. We built a prognostic model based on methylation. Then, we explored BTG2 expression and NSCLC survival in 3,038 cases, including the above-mentioned cohorts as well as 17 extended public datasets by meta-analysis. Further, we integrated the clinical information, expression, and methylation to build an integration model and evaluated its prediction ability for 5-year survival.

Results Three risk CpG probes (cg01798157, cg06373167, cg23371584) were associated with overall survival. The prognostic model could distinguish patient survival in the four cohorts [hazard ratio (HR) range, 1.51 to 2.21] and the independent validation set (HR = 1.85). In the expression analysis, BTG2 acted as a tumor-suppress gene in each cohort (HR range, 0.28 to 0.68). A meta-analysis showed high BTG2 expression was associated with better survival (HR = 0.61, 95%CI: 0.54-0.68). The three CpG probes were all negatively correlated with BTG2 expression. Additionally, the integration model showed a better prediction ability in the training set [area under the curve (AUC) = 0.78] and the validation set (AUC = 0.77). The integrated signature showed a better predictive ability among cases with adjuvant therapy (AUC = 0.85). In addition, 17 miRNAs including miR-21 and miR-23a were associated with BTG2 expression and survival.

Conclusions: methylation and integrated prognostic signatures of BTG2 are stable and reliable biomarkers for early-stage NSCLC. These markers may have new applications for appropriate clinical adjuvant trials and personalized treatments in the future.

Citation Format: Sipeng Shen, Li Su, Ruyang Zhang, Elizabeth Loehrer, Michael Lanuti, Nancy Diao, David Christiani. A multi-omic study reveals BTG2 as a reliable prognostic marker for early-stage non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-159.
Abstract LB-168: Inhibitory activities of Sesamum indicum L. leaf extract against the growth, invasion, migration, and adhesion of human colon cancer cells
Authors: Kim Seoyun , Ju Jihyeung
Published: August 2018

Colorectal cancer is one of the most common cancers in both incidence and mortality among men and women worldwide. Sesame (Sesamum indicum L.) leaves (SL) are used as vegetables and traditional medicines in Asian and African countries. In the current study, we aimed to investigate the effects of ethanol extract of sesame leaves (SLE) against key characteristics of cancer cells, including unrestricted growth, resisted apoptosis, and activated metastasis, and to evaluate whether thermal treatment of SL alter the inhibitory effects. Two human colon cancer cell lines were used in this study, HT29 colon adenocarcinoma cells and HCT116 colorectal carcinoma cells. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, treatment with SLE at the concentrations of 50-500 µg/mL for 24-72h resulted in dose-dependent and time-dependent growth inhibition in HT29 and HCT116 cells. In cell cycle analyses, SLE caused significantly increased cell population at sub-G1 (in HT29 and HCT116) and G2/M (in HCT116) phases, suggesting that the observed growth inhibition is associated with induction of apoptosis and cell cycle arrest. SLE at non-cytotoxic concentrations was effective in inhibiting critical events during metastasis in HT29 and HCT116, such as invasion (determined by matrigel-coated transwell assay), migration (determined by wound-healing assay), and adhesion (determined using fibronectin-coated plates). Thermal treatment of SL (at 150°C for 15 min) resulted in significantly enhanced inhibitory activities of SLE against the invasion, migration and adhesion in HCT116 cells, but did not change the growth-inhibitory and apoptosis-inducing activities. These results suggest that SLE possesses in vitro anti-cancer activities against colon cancers and that thermal treatment of SL enhances the anti-metastatic activities of SLE. Further studies are needed to verify whether similar effects are reproduced in vivo and to identify active ingredients of SLE responsible for the inhibitory activities.

Citation Format: Seoyun Kim, Jihyeung Ju. Inhibitory activities of Sesamum indicum L. leaf extract against the growth, invasion, migration, and adhesion of human colon cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-168.
Abstract LB-170: 9cis-retinoid UAB30 manifests potent chemopreventive effects in a murine model of UVB-induced basal cell and squamous cell carcinoma
Authors: Chaudhary Sandeep C. , Kashyap Mahendra P. , Nguyen Gloria , Elmets Craig A. , Athar Mohammad
Published: August 2018

Vitamin A derivatives such as retinoids are known to inhibit cell proliferation and induce cellular differentiation/apoptosis in various tumor cells. Although many retinoids have been limited due to their severe adverse effects, but 9cUAB30, a highly selective novel retinoid X receptors (RXR)-agonist has been shown an effective anti-carcinogenic activity. Here, we demonstrated the chemopreventive and chemotherapeutic efficacy of 9cUAB30 in UVB-induced photocarcinogenesis in Ptch+/-/SKH-1 murine model, which develops both basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). Both topical and systemic application to 9cUAB30 significantly reduced UVB-induced skin photocarcinogenesis in terms of tumor number and tumor volume. 9cUAB30 reduced UVB-induced skin hyperplasia, proliferation, and enhanced differentiation of suprabasal epidermal keratinocytes. Induction of apoptosis was also noted in these neoplastic lesions treated with this retinoid. 9cUAB30 was almost equally potent in the inhibition of either UVB-induced skin BCCs or SCCs in Ptch+/-/SKH-1 mice. 9cUAB30-treated residual tumors were manifesting reduced epithelial-mesenchymal transition (EMT) progression with a less aggressive tumor phenotype, which was associated with the enhanced expression of E-cadherin and reduced expression of N-cadherin, Fibronectin, Snail and Twist. 9cUAB30-mediated therapeutic intervention against these tumors was also involved induction of extensive DNA double strand breaks (DSBs) characterized by the enhanced phosphorylation of H2AX. This DNA damage response was unique to 9cUAB30 compare to other known RXR inhibitors such as Bexarotene. It was also independent of reactive oxygen species (ROS) production as ROS scavenger namely, N-acetyl cysteine did not effectively block it. However, 9cUAB30 provoked DNA damage response signaling was associated with the enhanced phosphorylation and activation of ATM, Rad50, ATR, Chk1, and Chk2. In summary, these data demonstrate that 9cUAB30 is a novel cancer chemopreventive agent for UVB-induced skin neoplasm and acts by invoking extensive DDSBs similar to those caused by chemotherapeutic drug like cisplatin.

Citation Format: Sandeep C. Chaudhary, Mahendra P. Kashyap, Gloria Nguyen, Craig A. Elmets, Mohammad Athar. 9cis-retinoid UAB30 manifests potent chemopreventive effects in a murine model of UVB-induced basal cell and squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-170.