bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5930: Oral siRNA delivery for colorectal liver metastases cancer therapy
Authors: Kang Sung-Hun , Revuri Vishnu , Cho Sungpil , Park In-Kyu , Cho Kwang Jae , Bae Woo Kyun , Lee Yong-Kyu
Published: August 2018

Liver metastasis (CLM) is the lethal secondary hepatic cancer with 5 - 20 months' average survival rate and mostly originated from colorectal cancer (CRC). Surgery and systemic chemotherapy are the currently best options to treat patients. However, over 60% of recurrence rate and severe side effects such as sinusoidal obstruction syndrome and nodular regenerative hyperplasia from the systemic chemotherapy significantly impact on patient's recovery from the treatment. To overcome these side effects and also achieve the synergistic therapeutic effect, we developed a dual padlock system (AR-GT) for oral siRNA delivery to regulate PI3K/Akt signaling pathway, which is highly associated with the progression of CRC to CLM. AR-GT is an about 130 nm nanoparticle consisted of a gold nanoparticle core anchoring siAkt RNA (AR) and a single outer layer of glycol chitosan-taurocholic acid (GT) conjugate. GT layer plays several roles in oral siRNA delivery such as the protection of siRNA in the gastrointestinal (GI) environment, facilitation of ileac uptake of AR-GT in the small intestine and transportation of AR-GT through enterohepatic circulation into the liver. In vitro characterization of AR-GT demonstrated the protection of siRNA under simulated GI pHs for 5 days, facilitation of the trans-epithelial transport in the intestinal Caco2 and ASBT positive cells, and reduction of Akt protein by successful release of siAKT from the AR-GT. Following oral delivery of AR-GT to CLM mouse model, we observed high biodistribution of AR-GT in the ileum, liver, and kidney through intestinal uptake, enterohepatic transportation, and renal excretion. Moreover, CLM mouse after oral delivery of AR-GT (100 μg/kg) manifested about 58 % reduction of the numbers of tumor nodules in the liver and near 100% survival benefit compared to control mouse fed with either saline or AR-Chitosan nanoparticles. Further analysis of liver samples obtained from CLM mouse after oral delivery of AR-GT indicated about 38% reduction of pAkt expression and the enhancement of apoptotic protein expressions such as 2 times or 1.5 times increase of caspase 9 and bax. Theses observation suggests survival benefit and tumor nodule reduction in CLM mouse root on successful AR-GT mediated oral siAkt RNA delivery. we conclude AR-GT oral siRNA delivery system is beneficial in the CLM treatment and further suggest its application to target other cancers.

Citation Format: Sung-Hun Kang, Vishnu Revuri, Sungpil Cho, In-Kyu Park, Kwang Jae Cho, Woo Kyun Bae, Yong-Kyu Lee. Oral siRNA delivery for colorectal liver metastases cancer 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 5930.
Abstract 597: Lung tumor histology as prognostic factor for short-term postoperative outcomes
Authors: Gerard Roger , Velez-Cubian Frank , Moodie Carla , Garrett Joseph , Fontaine Jacques , Toloza Eric
Published: August 2018

Introduction: Based on the 2015 World Health Organization (WHO) Classification of Lung Tumors, we sought to investigate the relationship of common lung tumor histologies and their utility as prognostic clinical correlates of short-term perioperative outcomes.

Methods: We retrospectively analyzed patients who underwent robotic-assisted video-thoracoscopic lobectomy by one surgeon over 81 months. Patients were grouped based on tumor histology on their final surgical pathology report. Patients' demographics, smoking history, lobar distribution, extent of resection, intraoperative outcomes, perioperative complications, and hospital length of stay were compared among the groups.

Results: Among 420 study patients, the most common tumor type was adenocarcinoma (AD, 59.8%), followed by squamous cell carcinoma (SQ, 19.3%) and neuroendocrine carcinoma (NEC, 11.7%). Final histology for the remainder of the cohort consisted of adenosquamous carcinoma (AdSq, 2.1%) and pulmonary metastasis (7.1%). All patients with AdSq tumor histology had a positive smoking history, which was a significantly higher proportion than for those with NEC tumor type (100% vs 71.4%; p<0.001). Those with AdSq tumor histology were both significantly older and developed significantly larger tumors by time of resection (p<0.001 and p=0.003, respectively). Patients with AdSq tumors required more extensive resections compared to the remaining groups (p<0.004). They also experienced significantly greater estimated blood loss (EBL) and significantly longer operative times (p=0.005 and p=0.018, respectively) than the remaining groups. Patients with SQ or pulmonary metastasis had slightly more postoperative complications, but this was not statistically significant (p=0.990).

Conclusion: Patients with AdSq histology type were older and had larger tumors, which put them at risk of intraoperative difficulties, with more extensive resections, longer operative times, and greater EBL. Despite this, AdSq histology did not affect the postoperative course.

Citation Format: Roger Gerard, Frank Velez-Cubian, Carla Moodie, Joseph Garrett, Jacques Fontaine, Eric Toloza. Lung tumor histology as prognostic factor for short-term postoperative outcomes [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 597.
Abstract 6: Role of Notch signaling in bevacizumab-induced vascular normalization in glioblastoma
Authors: Saito Norihiko , Aoki Kazuya , Hirai Nozomi , Fujita Satoshi , Nakayama Haruo , Hayashi Morito , Sakurai Takatoshi , Iwabuchi Satoshi
Published: August 2018

Tumor angiogenesis occurs in the setting of a defective vasculature, which is associated with increased vascular permeability and enhanced tumor permeability. Bevacizumab is used to treat malignant glioma and was found to reduce microvascular density and prune abnormal tumor microvessels. Using electron microscopic observation of two autopsy cases, we investigated the effects of blood vessel normalization in glioblastomas treated with bevacizumab. Notch-1 and SMA immunostaining were used to compare initial surgical specimens with postmortem specimens obtained after bevacizumab treatment. Postmortem samples showed marked proliferation of SMA-positive cells (pericytes) in tumor vessels and marked proliferation of Notch-1–positive cells around vessels. Electron microscopic images confirmed the presence of pericytes surrounding the vascular endothelium. These findings suggest that bevacizumab treatment promotes vascular normalization by recruiting mature pericytes. Next, we investigated the effects of bevacizumab on VEGF inhibition in glioma stem cells. Bevacizumab treatment attenuated activation of VEGFR2 and increased Notch signaling expression. VEGF inhibition by bevacizumab treatment attenuated proliferation and self-renewal of glioma stem cells and induced endothelial and pericyte differentiation. In tumor angiogenesis, vascular endothelial growth factor induces sprouting angiogenesis and recruitment of vascular endothelial cells such as tip cells, stalk cells, and phalanx cells. Fully mature phalanx cells are in close contact with pericytes. Our results suggest that bevacizumab treatment induces glioma stem cells to differentiate to endothelium and pericytes. These mechanisms might be important in normalizing tumor vasculature after bevacizumab treatment and could be useful in improving the effectiveness of current glioma therapy.

Citation Format: Norihiko Saito, Kazuya Aoki, Nozomi Hirai, Satoshi Fujita, Haruo Nakayama, Morito Hayashi, Takatoshi Sakurai, Satoshi Iwabuchi. Role of Notch signaling in bevacizumab-induced vascular normalization in glioblastoma [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 6.
Abstract 604: Metabolomics in predicting breast cancer treatment responses
Authors: Barcha Carolina Puyana , Lee Eunkyung , Takita Cristiane , Wright Jean L. , Hu Jennifer
Published: August 2018

Breast cancer is the most frequently diagnosed cancer in women and the second leading cause of cancer death in American women. Postoperative adjuvant radiotherapy (RT) significantly improve local-regional recurrence and survival. Therefore, there has been increasing usage of adjuvant RT in early-stage breast cancer patients. Although well tolerated by some patients, 48% patients in our study experience RT-induced early adverse skin reactions (EASRs) that negatively impact quality of life. Therefore, we evaluated urine metabolomics in predicting RT-induced EASRs and precision intervention. Based on the pilot data of 478 metabolites in 60 patients, we used the MetaboAnalyst 3.0 (www.metaboanalyst.ca) to conduct metabolomic data analysis, visualization and interpretation. Using KEGG pathway analysis, we have identified 7 pathways that are significantly associated with RT-induced EASRs (i.e., False Discovery Rate (FDR) p less than 0.05). The alanine, aspartate and glutamate metabolism pathway has the most significant FDR p-value and the highest impact value of 0.60 in predicting RT-induced EASRs. In summary, to the best of our knowledge, this is the first study in breast cancer patients associating glutamate metabolism pathway to RT-induced EASRs. Our research will have significant clinical impact because: (i) many tumor cells depend on glutamine-glutamate as energy source for growth and survival, (ii) breast cancer cells secrete high levels of glutamate often metastasize to bone with severe pain issue, (iii) glutaminase inhibitors enhance chemotherapy in triple negative breast cancer, (iv) two thirds of cancer patients receive RT, and (v) glutamate metabolism is associated with RT-induced EASRs. With increasing recent interest in targeting tumor metabolism, we anticipate that glutaminase inhibitors will also have application in preventing RT-induced EASRs in addition to their anticancer activities in triggering metabolic crisis in tumor cells and enhancing chemotherapy.

Citation Format: Carolina Puyana Barcha, Eunkyung Lee, Cristiane Takita, Jean L. Wright, Jennifer Hu. Metabolomics in predicting breast cancer treatment responses [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 604.
Abstract 629: Delivering type I interferon to dendritic cells empowers tumor eradication and immune combination treatments
Authors: Tavernier Jan H. , Cauwels Anje , Lint Sandra Van , Paul Franciane , Garcin Geneviève , Parys Alexander Van , Kley Nikolai , Uzé Gilles
Published: August 2018

An ideal generic cancer immunotherapy should mobilize the immune system to destroy tumor cells without harming healthy cells and remain active in case of recurrence. Furthermore, it should preferably not rely on tumor-specific surface markers, as these are only available in a limited set of malignancies. Despite approval for treatment of various cancers, clinical application of cytokines is still impeded by their multiple toxic side effects. Type I interferon (IFN), for instance, has a long history in the treatment of cancer, but its multifaceted activity pattern and complex side effects prevent its optimal clinical use. Here we develop AcTakines (Activity-on-Target cytokines), optimized (mutated) immunocytokines that are up to 1000-fold more potent on target cells, allowing specific signaling in selected cell types only. As conventional Dendritic Cells (cDC) are essential for IFN antitumor efficacy, we targeted type I IFN-derived “AcTaferon (AFN)” to Clec9A+ cDC. Clec9A-AFN therapy displayed strong antitumor activity in murine melanoma (B16), breast carcinoma (4T1) and lymphoma models (A20), as well as against human RL lymphoma in immunodeficient NSG mice reconstituted with a human immune system. In sharp contrast to wild-type IFN therapy, the antitumor efficacy of Clec9A-AFN was not accompanied by any detectable toxicity, assessed by body weight and several hematological parameters. Clec9A-AFN effects were lost in CD8-depleted or Batf3-/- mice, and depended on IFN signaling in cDCs but not in T lymphocytes. Combined with α-PDL1 immune checkpoint blockade, Treg-depleting anti-CTLA4 + anti-OX40 therapy, immunogenic chemotherapy, or low-dose TNF, complete tumor regressions and long-lasting tumor immunity (memory) were obtained, still without any adverse effects. Our findings thus indicate that DC-targeted AFN provides a highly efficient, off-the-shelf and safe cancer immunotherapy, with possible application in a broad range of malignancies.

Citation Format: Jan H. Tavernier, Anje Cauwels, Sandra Van Lint, Franciane Paul, Geneviève Garcin, Alexander Van Parys, Nikolai Kley, Gilles Uzé. Delivering type I interferon to dendritic cells empowers tumor eradication and immune combination treatments [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 629.
Abstract 633: Application of a novel analytical metric that quantifies copy number alterations in low-coverage, genome-wide sequencing of cell-free DNA to monitor and differentiate response to immunotherapy in cancer patients
Authors: Jensen Taylor J. , Goodman Aaron M. , Ellison Christopher K. , Kato Shumei , Daniels Gregory A. , Tran Lisa , Nakashe Prachi , McCarthy Erin , Mazloom Amin R. , McLennan Graham , Grosu Daniel S. , Ehrich Mathias , Kurzrock Razelle
Published: August 2018

Inhibitors of the PD-1/PD-L1/CTLA4 immune checkpoint pathway have transformed the cancer treatment paradigm. It has been observed that even some patients with advanced, refractory malignancies achieve durable responses; however, only a minority of patients benefit, demonstrating the importance of developing new biomarkers to predict patient outcome. The most commonly used biomarkers for predicting response to checkpoint inhibitors are the expression of PD-1/PD-L1, microsatellite instability, and tumor mutational burden. While these markers have been shown to have varying degrees of predictive power, they have not been used to monitor and differentiate response during treatment and require invasive procedures. Interrogating cell-free DNA (cfDNA) isolated from plasma (liquid biopsy) provides a promising noninvasive method for monitoring response. We describe the use of low-coverage (~0.3X) genome-wide sequencing of cfDNA, validated extensively for detecting chromosomal abnormalities in non-invasive prenatal testing (NIPT) and previously shown to enable incidental detection of occult maternal malignancies, to detect tumor-specific copy number alterations; and the development of a new metric–the genome instability number (GIN)–to monitor response to these drugs. Using a series of more than 450 plasma aliquots taken from more than 75 patients throughout their treatment, we demonstrate how the GIN can be used to discriminate clinical response from progression, differentiate progression from pseudoprogression, and identify hyperprogressive disease, as early as 4-6 weeks after treatment initiation. Finally, we provide evidence for a delayed therapeutic response to checkpoint inhibitors relative to targeted therapies. While this study is still ongoing, these initial data provide proof of concept for using this method for monitoring treatment outcome in cancer patients receiving immunotherapy.

Citation Format: Taylor J. Jensen, Aaron M. Goodman, Christopher K. Ellison, Shumei Kato, Gregory A. Daniels, Lisa Tran, Prachi Nakashe, Erin McCarthy, Amin R. Mazloom, Graham McLennan, Daniel S. Grosu, Mathias Ehrich, Razelle Kurzrock. Application of a novel analytical metric that quantifies copy number alterations in low-coverage, genome-wide sequencing of cell-free DNA to monitor and differentiate response to immunotherapy in cancer 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 633.
Abstract 64: Macrophage perceive tumor-intrinsic glutamine metabolism and orchestrate immunosuppression through IL-23 inclear cell renal cell carcinoma
Authors: Fu Qiang
Published: August 2018

The signaling mechanisms between renal cancer cell metabolism and infiltrating immune cells may illuminate novel therapeutic approaches. Here, we showed the idiomatic glutamine metabolism of clear cell renal cell carcinoma (ccRCC) suppress immune surveillance through nonautonomous IL-23. Through analyzing multiple ccRCC clinical cohorts and human tumor samples, we uncovered SLC1A5 (solute carrier family 1 member 5) promotes ccRCC consumption of extrinsic glutamine, predicts patient prognostication, and associates with reduction of cytotoxic T cells and NK cells in the tumor microenvironment. Specifically, the immune surveillance suppression of tumor-intrinsic SLC1A5 is considerably conducted by IL-23, which could directly induce intratumoral accumulation of T helper 17 cells and regulatory T cells. Intriguingly, we identified macrophages as the major source of intratumoral IL-23, and macrophages secrete large amount of IL23 under low glutamine condition, as the consequence of tumor-driven glutamine consumption. Furthermore, blocking IL-23 using specific antibody in syngeneic mice tumor model prolonged their survival, and blocking IL-23 in an in vitro culture model of human tumors promote anti-tumoral T and NK cells reaction. Our results demonstrate that macrophage-secreting IL-23 is a key immune effector as the response to tumor-driving glutamine metabolism, and IL-23 profoundly facilitate an immunosuppressive microenvironment which suggest it as a promising target for cancer immunotherapy in ccRCC.

Citation Format: Qiang Fu. Macrophage perceive tumor-intrinsic glutamine metabolism and orchestrate immunosuppression through IL-23 inclear cell renal 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 64.
Abstract 648: Digital PCR in genetic diagnosis of NUT midline carcinoma
Authors: Okumura Shunsuke , Chiba Shin-ichi , Sado Masatoshi , Takahashi Nana , Sasaki Takaaki , Kitada Masahiro , Ohsaki Yoshinobu
Published: August 2018

Background; NUT midline carcinoma (NMC) is a rare, highly aggressive tumor with t(15;19) translocation involving NUT midline carcinoma family member 1 (NUTM1) gene. NMC is genetically defined and diagnosed by RNA sequencing or by immunohistochemistry for NUT protein expression instead. However, it can be challenging to perform these assays on small biopsy specimens. We, here, report the utility of digital PCR (dPCR) assay for diagnosis of NMC. Methods; The QuantStudio 3D dPCR system (Thermo Fisher Scientific) was used. We designed optimal dPCR probes for the BRD4-NUT fusion gene and evaluated the dPCR method using NMC cell lines (HCC2429 and Ty82) and clinical samples. Total RNA was extracted from the cell lines and FFPE tissue samples from patients with thoracic tumors (RecoverAll total nucleic acid isolation kit, Thermo Fisher Scientific). cDNA was synthesized with SuperScript VILO cDNA kit (Thermo Fisher Scientific). PCR was performed using ProFlex PCR system (Thermo Fisher Scientific). Data was analyzed with QuantStudio 3D AnalysisSuite Cloud Software (Thermo Fisher Scientific). Results; We identified the BRD4-NUT fusion gene in HCC2429 and Ty82 cells using the dPCR system, which allowed us to detect the fusion gene at a prevalence < 0.1%. The fusion genes were detected in the clinical samples, and we successfully found NMC patients using the dPCR assay. Conclusion; We conclude the dPCR assay is an appropriate method for diagnosis of NMC.

Citation Format: Shunsuke Okumura, Shin-ichi Chiba, Masatoshi Sado, Nana Takahashi, Takaaki Sasaki, Masahiro Kitada, Yoshinobu Ohsaki. Digital PCR in genetic diagnosis of NUT midline 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 648.
Abstract 649: Development of a novel RNA sequencing approach that identifies aberrant splicing in cancer predisposing genes
Authors: Jansen Anne M. , Klift Heleen M. van der , Eendenburg Jaap D. van , Tops Carli M. , Wijnen Juul T. , Hes Frederik J. , Goel Ajay , Morreau Hans , Wezel Tom van
Published: August 2018

Background: High-throughput sequencing efforts in molecular tumor diagnostics detect increased numbers of novel genetic variants, including the ones that are predicted to affect splicing. Current approach for predicting whether a specific variant may result in aberrant RNA transcripts often consists of in-silico prediction using bioinformatics prediction tools, which is sometimes followed by RT-PCR analysis of the RNA extracted from blood, or functional splicing reporter assays. Although high-quality patient RNA analysis is usually preferred, such RNA is not always available, or the analysis is hampered due to degradation of aberrant transcripts through nonsense-mediated decay. To overcome this issue, we investigated the possibility of analyzing RNA isolated from formalin-fixed paraffin-embedded (FFPE) tissues, with the underlying hypothesis that formalin fixation may inhibit RNA degradation, enabling the detection of aberrant RNA products. Methods: RNA analysis was performed for 13 canonical splice site variants predicted or known to affect splicing in the cancer predisposition genes MLH1, MSH2, MSH6, APC and BRCA1. Of 11 variants, total nucleic acid was obtained from FFPE tumor tissues. For two variants, no FFPE material was available, but EBV-transformed B-cells carrying a splice variant were cultured. Primers were designed to amplify the exon-exon boundary predicted to be affected. Housekeeping gene expression was assessed to determine presence of RNA. Results: Total nucleic acid was successfully isolated for 10 variants from eight FFPE tumor tissues and two EBV-transformed B-cell lines. Aberrant splicing was confirmed in all six variants known to result in splicing. Of one known splice variant, present in the B-cell line, the known aberrant splice product could only be detected after formalin fixation of the cells, suggesting that formalin fixation possibly inhibited RNA degradation. Conclusion: Aberrant splicing was successfully identified in all known splice variants and three of four variants predicted to affect splicing, of which the splice effect was previously unknown. Using a RNA sequencing approach, somatic splice variants could be easily and rapidly analyzed, enabling retrospective analysis of variants predicted to result in splicing when only FFPE material is available in clinical settings.

Citation Format: Anne M. Jansen, Heleen M. van der Klift, Jaap D. van Eendenburg, Carli M. Tops, Juul T. Wijnen, Frederik J. Hes, Ajay Goel, Hans Morreau, Tom van Wezel. Development of a novel RNA sequencing approach that identifies aberrant splicing in cancer predisposing genes [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 649.
Abstract 65: Impact of tumor-infiltrating dendritic cells in esophageal squamous cell carcinoma treated with neoadjuvant chemotherapy
Authors: Nishimura Junya , Tanaka Hiroaki , Yamakoshi Yoshihito , Hiramatsu Soichiro , Tamura Tatsuro , Toyokawa Takahiro , Muguruma Kazuya , Hirakawa Kosei , Ohira Masaichi
Published: August 2018

Background: Tumor-infiltrating CD8+T cells have been found to be associated with favorable outcome of patients in many cancers including esophageal squamous cell carcinoma (ESCC). Dendritic cells (DCs) are the most potent antigen-presenting cells to induce CD8+T cells. Several investigators have shown that tumor infiltrating mature DCs (mDCs) were associated with favorable prognosis of cancer patients. We showed that mDCs were associated with favorable prognosis of ESCC patients who had not received preoperative treatment. Neoadjuvant chemotherapy (NAC) is the standard treatment for Stage II/ III thoracic esophageal carcinoma in Japan. The effect of chemotherapy on tumor infiltrating DCs remains unclear.

Purpose: The aim of this study was to examine the correlation of efficacy of NAC and locally infiltrating DCs in ESCC patients.

Object and Methods: Formalin-fixed, paraffin-embedded blocks of primary lesions were collected from 50 patients with ESCC who underwent surgical treatment after NAC at Osaka City University Hospital. Immunohistochemical analysis using anti-LAMP-3 antibody and CD8 antibody was used to evaluate tumor infiltrating DCs and CD8+T cells. The number of LAMP-3+DCs and CD8+T cells were counted at ×400 magnification. We divided the 50 cases into two groups according to the median number of tumor infiltrating DCs and CD8+T cells to examine the correlation of efficacy of NAC and locally infiltrating DCs.

Results: LAMP-3+DCs were predominantly observed in peritumoral area and sparsely found in intratumoral area, whereas CD8+T cells distributed between both peritumoral and intratumoral areas. LAMP-3+DCs infiltrated the peritumoral space from 0 to 19.0 (median 6.0) and the intratumoral space from 0 to 3.8 (median 0.0). Patients with histological effect (≧Grade 1a) had higher infiltration of mDCs than Grade 0. Patients with high infiltration of mDCs had better prognosis than those with low. The number of mDCs had positive correlation with the number of tumor infiltrating CD8+T cells.

Conclusions: Our results suggested that increase of tumor infiltrating mDCs was associated with histological effect of NAC and favorable prognosis in ESCC.

Citation Format: Junya Nishimura, Hiroaki Tanaka, Yoshihito Yamakoshi, Soichiro Hiramatsu, Tatsuro Tamura, Takahiro Toyokawa, Kazuya Muguruma, Kosei Hirakawa, Masaichi Ohira. Impact of tumor-infiltrating dendritic cells in esophageal squamous cell carcinoma treated with neoadjuvant chemotherapy [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 65.
Abstract 660: Integration of radiomics with "omic" analyses to predict survival in newly diagnosed IDH-1 wild-type glioblastoma
Authors: Daniel Paul M.
Published: August 2018

Glioblastoma (GBM) is the most commonly diagnosed glioma and has the poorest median survival of all brain tumours at only 14 months. Despite intensive characterisation of this disease through ‘omic' investigations which has led to the identification of critical molecular drivers of malignancy, only a limited selection of biomarkers have proven to be clinically relevant in predicting patient survival. One reason for this lack of success is the existence of intratumoural heterogeneity, where individual tumours can harbour spatially separated molecular and phenotypic diversity, thereby limiting the insight which ‘omic' analysis of small biopsy specimens can impart. Radiomics involves high-throughput mining of quantitative image features via image acquisition, region of interest segmentation and feature extraction from standard MRI scans and relating extracted feature values to molecular, phenotypic and clinical properties. Notably, radiomics derives features from the whole tumour mass and therefore overcomes limitations associated with intratumoural heterogeneity. As such, we sought to investigate the ability for radiomics to describe tumour characteristics and integrate radiomics alongside genomics and transcriptomics to derive a prognostic model for IDH1 wild-type GBM. We observed 2 radiomic clusters in IDH1-wild-type GBM which each had an equivalent distribution of each of the four molecular subtypes. Whilst radiomic clusters themselves were not predictive of survival, three radiomic features were able to separate patients into long and short survival groups. Integration of genomic and transcriptomic features alongside radiomic analysis increased the predictive strength of our model. Finally, we found an association between longer survival in patients with IDH1 wild-type GBM and increased expression of the complement system. Our observations suggest that integration of radiomics alongside ‘omic' investigations offer a greater ability to predict survival in IDH1 wild-type GBM and suggests a role for the innate immune system in driving patient outcome.

Citation Format: Paul M. Daniel. Integration of radiomics with "omic" analyses to predict survival in newly diagnosed IDH-1 wild-type glioblastoma [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 660.
Abstract 662: IMA_Detect: Mass spectrometry guided development and clinical application of a companion diagnostic for adoptive cellular therapy against tumor associated HLA peptides
Authors: Fritsche Jens , Satelli Arun , Hörzer Helen , Rakitsch Barbara , Hoffgaard Franziska , Hilf Norbert , Schoor Oliver , Singh-Jasuja Harpreet , Toni Weinschenk
Published: August 2018

Adoptive cellular therapy (ACT) has dramatically changed the landscape of cancer immunotherapy. ACTolog® and ACTengine® are actively personalized ACT approaches employing T-cell receptor products based on a warehouse of human leucocyte antigen (HLA)-bound peptide targets. Selecting the relevant target candidates requires the establishment of biomarkers predictive for HLA peptide presentation and their development into companion diagnostic devices.

Here we describe the development of IMA_Detect, a diagnostic test based on gene expression analysis of primary patient tumor material by qPCR which is predictive for presentation of a target peptide by HLA. To establish mRNA expression levels which indicate actual peptide presentation, data of Immatics' XPRESIDENT® target discovery platform was used by integrating quantitative immunopeptidomics data (label-free LC-MS) with paired transcriptomics data (RNA-Seq). The peptide-specific correlation between peptide presentation and expression of the coding exons was verified for each target individually. The resulting RNA-Seq thresholds were mapped to qPCR thresholds using calibration curves followed by validation of the qPCR assay.

The IMA_Detect test is performed in a CLIA/CAP approved setting and is currently being applied in the phase I ACT trials IMA101-101 and IMA201-101 to determine whether a patient's tumor expresses any of the tested targets at levels considered sufficient for potential benefit from the administered T-cell therapy. IMA101-101 uses autologous T-cell products (ACTolog®) for patients with solid cancers while IMA201-101 is based on TCR-engineered T Cells (ACTengine®) in NSCLC and HNSCC patients. We will present first results of the patient screening and the personalized target selection.

Citation Format: Jens Fritsche, Arun Satelli, Helen Hörzer, Barbara Rakitsch, Franziska Hoffgaard, Norbert Hilf, Oliver Schoor, Harpreet Singh-Jasuja, Weinschenk Toni. IMA_Detect: Mass spectrometry guided development and clinical application of a companion diagnostic for adoptive cellular therapy against tumor associated HLA peptides [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 662.
Abstract 678: Raf-1 cysteine-rich domain (CRD) supports active orientations of KRas4B/Raf-1 at the membrane
Authors: Jang Hyunbum , Nussinov Ruth
Published: August 2018

Ras is a small GTPase, controlling signal transduction pathways and promoting cell proliferation and survival. In cancer, there are two independent pathways in tumor proliferation: one includes Raf/MEK/ERK (MAPK) and PI3K/Akt/mTOR. KRas4B is the most abundant oncogenic isoform. In the MAPK pathway, KRas4B preferentially recruits Raf-1 and activates it. The high-affinity interaction of Ras binding domain (RBD) of Raf with Ras was solved, but the relative position of Raf's cysteine-rich domain (CRD) in the Ras/Raf complex at the membrane and the key question of exactly how it affects Raf signaling are daunting. Here, we employ all-atom MD simulations to investigate membrane-anchored Raf-1 CRD and suggest a model of KRas4B/Raf-1 complex at the anionic lipid bilayer composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl-sn-glycero-3-phosphoserine (DOPS) in mole ratio 4:1. Active Ras modulates Raf activation, with KRas4B preferentially activating Raf-1. We show that the Raf-1 CRD stably binds anionic lipid bilayers inserting a positively-charged loop into the amphipathic interface. The loop contains key basic residues, responsible for the membrane association. Importantly, when in complex with Ras/RBD, covalently-connected CRD presents the same membrane interaction mechanism, with CRD locating at the space between the RBD and membrane, suggesting that CRD is an intrinsic membrane binding domain of Raf kinase. To date, CRD's role was viewed in terms of stabilizing Raf-membrane interaction. Our observations argue for a key role of CRD that Raf-1 CRD not only offers an additional anchor for the KRas4B/Raf-1 complex, but by reducing the fluctuations of Ras/RBD it also increases the population of Ras/RBD at the membrane and enhances the already high affinity between Ras and RBD. In the KRas4B/RBD-CRD complex, Raf-1 CRD supports the active-state Ras orientation at the membrane, releasing the catalytic domain from the transiently confined orientation. This promotes MAPK signaling, the key Ras proliferative pathway, above other pathways. Even though in the absence of CRD the Ras/Raf-1 RBD interaction can elicit MAPK signaling, the enhanced stability of the complex at the membrane rendered by CRD can considerably intensify it, by helping in Raf's dimerization. Our data reveal these significant roles of CRD at the membrane in the Raf activation. Funded by Frederick National Laboratory for Cancer Research, National Institutes of Health, under contract HHSN261200800001E.

Citation Format: Hyunbum Jang, Ruth Nussinov. Raf-1 cysteine-rich domain (CRD) supports active orientations of KRas4B/Raf-1 at the membrane [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 678.
Abstract 680: FLLL12 is a small molecule JAK2 inhibitor that inhibits JAK-STAT3 pathway in head and neck cancer
Authors: Amin ARM Ruhul , Anisuzzaman ASM , Siddique Abu B. , Fuchs James R.
Published: August 2018

Purpose: We have recently reported that FLLL12, a synthetic curcumin analog, induces apoptosis of lung cancer cells through a death receptor (DR) 5-dependent pathway and in squamous cell carcinoma of head and neck (SCCHN) by modifying multiple Bcl-2 proteins. However, the direct molecular target (receptor) of FLLL12 is unknown. The purpose of the current study is to identify the receptor of FLLL12.

Methods: Computer-aided molecular docking was used to predict molecular interaction. Isothermal titration calorimetry and thermal shift assays were used to confirm molecular interactions in a cell free system. SCCHN cell lines were used to study the effect of this interaction on JAK-STAT3 pathway. Protein expression levels were measured by Western blotting.

Results: Computer-aided molecular docking modeling of FLLL12 and JAK2 (PDB code: 5CF5) predicted that FLLL12 fit well into the JAK2 catalytic pocket. Although curcumin also fit into the JAK2 catalytic pocket, the binding sites were different for FLLL12 and curcumin. Docking score for FLLL12, natural ligand and curcumin were -7.2, -12.1 and -6.9, respectively. FLLL12 also docked into the STAT3 catalytic pocket (PDB code: 5AX3). However, the docking score was worse than those for natural ligand and curcumin (-5.1, -7.3 and -7.4 respectively). We confirmed that FLLL12 interacted with JAK2 in a cell free system using recombinant JAK2. Isothermal titration calorimetry assay revealed one binding site for both FLLL12 and the FDA-approved JAK inhibitor ruxolitinib. The Ka values were 3.84E6±1.03E5 and 2.31E7±6.78E6 M-1, respectively. FLLL12 and JAK2 interactions were further confirmed by thermal shift assay. Kd values were 1.12±0.14 μM and 2.24 ±0.18 μM for ruxolitinib and FLLL12, respectively. Two hour pretreatment with FLLL12 also inhibited IL-6 induced phosphorylation of JAK2 and STAT3. Finally, FLLL12 strongly inhibited constitutively active STAT3 phosphorylation in SCCHN cell lines, a downstream effector of JAK.

Conclusions: Our results strongly suggest that FLLL12 is a novel small molecule inhibitor for JAK2 and FLLL12-JAK2 interaction inhibits IL-6-induced as well as constitutively active STAT3 phosphorylation, thus has great promise for cancer prevention and therapy. Grant Support: Start-up fund from Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory University and from Department of Pharmaceutical Sciences and Research, Marshall University.

Citation Format: ARM Ruhul Amin, ASM Anisuzzaman, Abu B. Siddique, James R. Fuchs. FLLL12 is a small molecule JAK2 inhibitor that inhibits JAK-STAT3 pathway in head and neck 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 680.
Abstract 688: Triphenylstannyl 2-(benzylcarbamoyl)benzoate (Ch-319) modulates AKT/FOXO3a signaling and inhibits progression of prostate cancer
Authors: Waseem Durdana , Syed Deeba N. , Khan Gul Majid , Haq Ihsan-ul , Mukhtar Hasan
Published: August 2018

Prostate cancer (PCa) is the most commonly diagnosed non-cutaneous malignancy in men in the United States. Moreover, its incidence is predicted to surge in many parts of the world due to increase in aging population and change in dietary habits. The PTEN/AKT axis plays a critical role in regulating PCa growth and survival. Synthetic and natural compounds have been explored in preclinical models as potential inhibitors of AKT pathway that can suppress proliferation or induce apoptosis in PCa cells. Triphenyltin compounds are complexes of tin with three phenyl groups attached to hydrocarbon ligands. Structural modifications have been attempted to yield organotins with enhanced cytotoxicity against cancer cells. Here, we synthesized a series of triphenyltin (IV) carboxylate derivatives and evaluated their efficacy against PCa. From this library, triphenylstannyl 2-(benzylcarbamoyl)benzoate (Ch-319) was selected for further studies on the basis of its selective cytotoxicity towards cancer cells. Treatment of PTEN-null PC3M and DU145 PCa cells with Ch-319 (250-500 nM) resulted in reduced viability and induction of cell cycle arrest associated with downregulation of PI3K and dephosphorylation of AKT and PRAS40. In parallel, elevation of FOXO3a expression, activation of pro-apoptotic Bax and downregulation of anti-apoptotic Bcl2 protein was observed. Flowcytometry revealed increased population of apoptotic cells in Ch-319 treated cells. Presensitization of PCa cells with Ch-319 augmented the effect of cabazitaxel, a commonly used taxane in patients with castration-resistant PCa. We next evaluated the effect of Ch-319 in a prostate-specific Pten-/- model where AKT activation is the major driving force for development of prostate adenocarcinoma. Mice treated with Ch-319 (5 µg/animal:intraperitoneally) for 6 weeks showed decreased tumor weights as compared to DMSO treated controls. Histological studies indicated a decrease in the number and size of foci in Ch-319 treated tumors. Consistently, reduced expression of androgen receptor, decreased phosphorylation of AKT and PRAS40 and increase in FOXO3a was observed in Ch-319 treated animals with no signs of organ toxicity. Our studies identify AKT/FOXO3a signaling as a target of triphenyltin (IV) carboxylate derivative Ch-319 and provide a molecular basis of its growth inhibitory effect in PCa cells. We propose that Ch-319 has potential to be developed as an anticancer agent against PCa.

Citation Format: Durdana Waseem, Deeba N. Syed, Gul Majid Khan, Ihsan-ul Haq, Hasan Mukhtar. Triphenylstannyl 2-(benzylcarbamoyl)benzoate (Ch-319) modulates AKT/FOXO3a signaling and inhibits progression of prostate 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 688.
Abstract 689: Identification and optimization of novel chemical matter via a structure-based approach resulting in a probe for MTH1
Authors: Bauser Marcus , Giese Anja , Ellermann Manuel , Guenther Judith , Eheim Ashley , Bunse Stefanie , Neuhaus Roland , Weiske Joerg , Quanz Maria , Glasauer Andrea , Nowak-Reppel Katrin , Bader Benjamin , Irlbacher Horst , Meyer Hanna , Queisser Nina , Haegebarth Andrea , Gorjanacz Matyas , Tresaugues Lionel , Ginman Tobias , Rahm Fredrik , Andersson Martin , Ericsson Ulrica , Forsblom Rickard , Lindstroem Johan , Silvander Camilla , Vicklund Jenny
Published: August 2018

Cancer cells can form reactive oxygen species (ROS) due to altered redox regulation that affect desoxynucleosides triphosphates (dNTP) in particular. 8-oxo-2'-deoxyguanosine-5'-triphosphate (8-oxo-dGTP) and 2-hydroxydeoxyadenosine-5'-triphosphate (2-OH-dATP) are the two most abundant oxidative nucleotide lesions in this respect. These undesired nucleoside triphosphates are sanitized by the hydrolase MTH1 (also known as NUDT1) in order to prevent their incorporation into replicating DNA. Sprint Bioscience created a series of drug-like, potent and selective MTH1 inhibitors using fragment-based drug discovery methods. In collaboration with Bayer, these inhibitors were extensively profiled, both in vitro and in vivo, to allow for the selection of a probe molecule with attractive properties for in vivo target validation studies. Herein, we would like to share novel chemical matter and it's binding to MTH1 in protein co-crystal structures. Furthermore, we describe the consecutive, stepwise structure-based optimization process. Extensive SAR elaboration clearly revealed the essential moieties for high potency and favorable ADME properties. We are able to report for the first time how we identified BAY-707 as a very potent and highly selective MTH1 inhibitor representing a potential probe to further evaluate the scope and limitations of MTH1 inhibition for therapeutic applications.

Citation Format: Marcus Bauser, Anja Giese, Manuel Ellermann, Judith Guenther, Ashley Eheim, Stefanie Bunse, Roland Neuhaus, Joerg Weiske, Maria Quanz, Andrea Glasauer, Katrin Nowak-Reppel, Benjamin Bader, Horst Irlbacher, Hanna Meyer, Nina Queisser, Andrea Haegebarth, Matyas Gorjanacz, Lionel Tresaugues, Tobias Ginman, Fredrik Rahm, Martin Andersson, Ulrica Ericsson, Rickard Forsblom, Johan Lindstroem, Camilla Silvander, Jenny Vicklund. Identification and optimization of novel chemical matter via a structure-based approach resulting in a probe for MTH1 [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 689.
Abstract 706: In vivo tumor-associated mutation screen identifies PI3K activation as a mechanism of resistance to PD-1 blockade
Authors: Collins Natalie B. , Miller Brian C. , Bi Kevin , Abosy Rose Al , Yates Kathleen , Shrestha Yashaswi , Doench John , Boehm Jesse , Haining W Nicholas
Published: August 2018

Known genomic correlates of response to immunotherapy do not perfectly predict clinical outcome, supporting the existence of unknown mechanisms of resistance to tumor immunity. We hypothesize that somatic, cancer-associated mutations account for heterogeneity in spontaneous response to tumors and response to immunotherapy. We have undertaken a systematic in vivo screen to identify mechanisms of resistance to tumor immunity in order to define a comprehensive set of therapeutic targets and provide biomarkers of sensitivity to immunotherapy. Mouse tumor cell lines (MC38 colon carcinoma or B16 melanoma) were engineered to express a library of barcoded open reading frames (ORFs) mutagenized to encode known cancer-associated somatic mutations. Tumor-bearing animals were treated with anti-PD-1. A mutation in Phospho-Inositol 3 Kinase (PI3K), PIK3CA c.3140A>G, increased in representation in anti-PD-1 treated tumors but not in immunodeficient animals, suggesting that activity of the mutant allele conferred selective growth advantage in the setting of tumor immunity. This mutation encodes a constitutively active mutant catalytic domain, PIK3CA H1047R. MC38 tumors homogeneously expressing H1047R and implanted into wild-type mice failed to respond to anti-PD-1 therapy, while tumors expressing a control gene regressed. Pharmacologic PI3K inhibition resensitized tumors to treatment with anti-PD-1. PD-1-treated PIK3CA H1047R tumors had fewer infiltrating CD8+ T cells as measured by immunohistochemistry and flow cytometry. Single-cell RNA-seq of tumor-infiltrating immune cells revealed a population of myeloid cells expressing known immune inhibitory proteins that differentially enriched in PIK3CA H1047R-expressing tumors. Our data suggest that PI3K has, in addition to its well-described oncogenic role, a role in tumor immune evasion mediated by establishment of an inhibitory myeloid microenvironment. As such, activating mutations in PI3K may be useful as a biomarker of poor response to immunotherapy, and these studies provide a rationale for therapeutic combination trials of PI3K inhibition with checkpoint blockade and other myeloid-targeting immunotherapies.

Citation Format: Natalie B. Collins, Brian C. Miller, Kevin Bi, Rose Al Abosy, Kathleen Yates, Yashaswi Shrestha, John Doench, Jesse Boehm, W Nicholas Haining. In vivo tumor-associated mutation screen identifies PI3K activation as a mechanism of resistance to PD-1 blockade [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 706.