bluuebunny/crossref_metadata_2025 huggingface dataset:
STING-dependent preventive effect by ablative hyperfractioned radiotherapy and delayed delivery of a CSF-1R inhibitor.
Authors: Sun Xiaowen , Deng Lei , Yin Limei , Li Mengqian , Xue Jianxin , Lu You
Published: May 2019

e14190

Background: It has been reported that hyperfractioned radiotherapy can bring out abscopal effects when combined with checkpoint inhibitors and radiation-induced myeloid-derived suppressor cells (MDSCs) infiltration can lead to radioresistance. Our prior work has shown that ablative hypofractionated radiotherapy (AHFRT) alone can not trigger significant abscopal effects in three mouse tumor models. The goal of this study was to figure out whether the AHFRT play a preventive role in different tumor models and can be enhanced by a CSF-1R inhibitor. Methods: In experiment exploring the preventive effect of AHFRT, on Day 0, B16/ CT26/ MC38 tumor cells or PBS were inoculated s.c. in the left-side thigh of mice, respectively. Then, control mice and mice with established tumors were irradiated by AHFRT (12Gy*3f*3days). When the AHFRT accomplished, mice randomly divided into different groups and the second tumors inoculated s.c. in the right-side thigh, respectively: 1 day/ 3 days/ 7 days after AHFRT accomplished. Besides, “1 day after AHFRT accomplished” mice, both established MC38/CT26 tumors and PBS controls, were divided into 4 groups: control, a-CSF-1R, AHFRT, AHFRT+a-CSF-1R. And MC38 tumor cells were irradiated. The mice and cells followed for tumor growth and further analyses (mainly flow cytometry, western-blotting and qPCR). Results: We found that AHFRT alone could trigger significant preventive effects in” 7 days after AHFRT accomplished” group in B16 tumor model and “ 1 day/ 3 days/ 7 days after AHFRT accomplished” groups in CT26/ MC38 tumor models. In CT26/ MC38 tumor models, all the second tumors were cured which inoculated in “7 days after AHFRT accomplished” groups because of the preventive effects. And these cured mice keep survival without tumors when rechallenged at 180 days after AHFRT accomplished. AHFRT induced CD11b+CD11c+MHC-II+ antigen presenting cells at the 1st day after AHFRT accomplished and effector CD8+ T cells at 3rd day after AHFRT accomplished. AHFRT could active cGAS, STING, CXCL10 and IRF3 in the irradiated MC38 cells. And with a STING inhibitor, the growth of second tumors lost the preventive effect from AHFRT. And a delayed delivery of a small-molecule CSF-1R inhibitor decreased monocyte-MDSC(CD11b+CD11c-Gr1-) and delay the growth of second tumors inoculated 1 day after AHFRT accomplished. Conclusions: We show thatAHFRT triggers in situ vaccination and subsequent prevention effect. The prevention effect is STING-dependent and can be enhanced by a delayed delivery of CSF-1R inhibitor.
Introduction of a variable electrostatic field for targeted drug application following intraperitoneal aerosol chemotherapy (PIPAC).
Authors: Khosrawipour Veria , Chaudhry Haris , Mikolajczyk Agata , Schubert Justyna , Pigazzi Alessio , Khosrawipour Tanja
Published: May 2019

e14211

Background: In the treatment of Peritoneal Metastasis, the delivery of pressurized chemo-aerosols into the capnoperitoneum has been reported to show superior qualities to the application of conventional liquid chemotherapy. However, there is little basic knowledge on how aerosol particle motion within the abdominal cavity may be influenced and directed by means of an electrostatic field. This study aims to investigate aerosol motion within the abdominal cavity using an ex-vivo model. Material and Methods: All experiments were performed in an ex-vivo box as well as on postmortem swine abdomen, both established ex-vivo PIPAC models previously introduced by our research team. Extra-abdominally created, unipolar charged aerosols are introduced to the abdominal cavity. A modulated electrostatic field within the abdominal cavity is established to examine aerosol particle collision via fluorescence microscopy and photography. Results: Aerosol particle collision was observed on the peritoneum in the varying electric field. Preliminary data suggests that tissue penetration rates change with a varying electric field (p < 0.05). Drug distribution pattern showed a significant improvement with a varying electric field vs. without (p < 0.05). Conclusions: Our data suggest that extra-abdominally created, highly drug concentrated aerosol particles have the capacity to achieve high drug penetration rates in the peritoneum. The addition of a varying electric field enhances particle collision as well as significantly improves drug distribution patterns. Further studies are required to investigate the in-vivo safety and efficacy of a varying electric field and the significance of extra-abdominally created chemo-aerosols with regards to possible clinical applications.
Some characteristics of cancer-testis gene expression regulation in colorectal cancer.
Authors: Soldatova Kristina I. , Kit Oleg Ivanovich , Tolmakh Roman E. , Kutilin Denis S.
Published: May 2019

e14233

Background: Cancer-testis antigens (CTA) can be used in immunotherapy and for early detection of cancer. Despite numerous studies of the CTA expression in different tumors, their transcriptional activity and its regulation in colorectal cancer (CRC) remain poorly understood. The purpose of the study was to analyze the expression of cancer-testis genes (CT-genes) and mechanisms of its regulation in CRC. Methods: Tumor and intact tissues of the colon were studied in 60 patients. RNAs were isolated using the method described by Chomczynski and Sacchi (2006). The REVERTA-L reagent kit was used for the cDNA synthesis. Expression of 16 CT-genes (MAGE-A1, -A2, -A3, -A4, MAGE-B1, -B2, GAGE-1, -3, -4, MAGEC1, BAGE, XAGE3, NYESO1, SSX2, SCP1, PRAME1) and expression and copy number of 3 DNA methyltransferase genes (DNMT1, DNMT3A, DNMT3B) were determined by Real-Time qPCR (GAPDH and GUSB - reference genes). For the cluster analysis, we used our R scripts. Differences were assessed by the Mann-Whitney test, and correlations – by the Spearman's rank correlation coefficient (r). Results: The expression of 2 CT genes, SSX2 and PRAME1, was increased (p < 0.05) by 1.8 and 2.9 times, respectively, and the BAGE expression was decreased by 2.4 times in tumor tissues, compared to the normal tissues. The expression and copy number of DNMT3A in tumor tissues was 1.5 and 2 times higher (p < 0.05), and that of DNMT3B – 2 times lower (p < 0.005), compared to normal tissues. The copy number and expression of the DNMT1 gene did not change. A strong positive correlation (r = 0.996) between the expression and copy number of DNA methyltransferase genes was observed. Using cluster analysis (Hierarchical Clustering, Euclidean distance), we detected two clusters of CRC samples different in the expression of CT-genes and methyltransferases. Cluster 1 showed increased expression of DNMT1, DNMT3A and DNMT3B and decreased expression of BAGE, SSX2 and PRAME1; cluster 2 – decreased expression of DNMT1, DNMT3A and DNMT3B and increased expression of BAGE, SSX2 and PRAME1. Conclusions: The detected aberrant expression of the CT-genes BAGE, SSX2, PRAME1 in CRC depends on the expression of DNMT1, DNMT3A, DNMT3B, which in its turn depends on the copy number of the corresponding DNA methyltransferase genes.
Interleukin-2 boosts local and abscopal antitumor effect of radiotherapy combined with anti-PD-1: A translational research from clinical radio-memory effect.
Authors: Chen Dawei , Song Xinyu , Yue Jinbo , Xing Ligang , Yu Jinming
Published: May 2019

e14244

Background: Preclinical and clinical studies have shown that prior receipt of radiotherapy enhances antitumor immune responses, a phenomenon we call the “radio-memory effect.” However, all of the evidence regarding this effect to date comes from work with anti-PD-1 or anti-PD-L1 inhibitors. Here we explore whether this effect also occurs with other forms of immune therapy, specifically interleukin-2 (IL-2). If so, whether this triple combination therapy could produce synergetic anti-tumor effect. Methods: 1. Clinical work: we retrospectively assessed outcomes in patients with malignant peritoneal effusion (MPE) who had previously received radiotherapy for colon or rectum cancer within 18 months before the intrapleural infusion of IL-2 or cisplatin. All patients received intrapleural infusion of IL-2 or cisplatin, and most had had several cycles of standard chemotherapy for colon or rectum cancer. 2. Translational work: MC38 cell line (5×106) was subcutaneously inoculated into the right lower extremity of C57/BL mice to construct a primary tumor model. Six days later, MC38 cell line (1×106) was subcutaneously inoculated into the left lower limb to construct a second tumor model. The main research is as follows: To explore the optimal radiation dose and fraction for radiotherapy; Whether the addition of IL-2 will further enhance the efficacy of radiotherapy combined with anti-PD-1; To explore the optimal combination model of radiation, anti-PD-1 and IL-2; Whether the addition of IL-2 will increase the abscopal effect produced by radiotherapy and anti-PD-1. Results: 1. We identified 3,747 patients with MPE (median age 66 years [range 24–81)) treated at one of several institutions from August 2009 through May 2016; 412 patients had been treated with IL-2 and 592 with cisplatin and had survived for at least 3 months afterward. 2. Hypofractionated radiotherapy (12Gy × 2) combined with IL-2 is better; IL-2 will enhance the efficacy of radiotherapy combined with anti-PD-1; IL-2 and anti-PD-1 followed by radiotherapy might maximize the anti-tumor effect; IL-2 might increase the abscopal effect produced by radiotherapy combined with anti-PD-1. Conclusions: We speculate that previous radiotherapy could enhance the efficacy of IL-2 and the anti-tumor effect produced by radiotherapy combined with anti-PD-1 could be enhanced by IL-2. The “radio-memory” effect could be beneficial in future studies.
Early response assessment through multiparametric MRI based endpoints in a phase II multicenter study evaluating the efficacy of DPX-Survivac, intermittent low dose cyclophosphamide (CPA) and pembrolizumab combination study in subjects with solid tumors.
Authors: Roettger Diana , Shaikh Faiq , Bisdas Sotirios , MacDonald Lisa Diana , Fiset Stephan , Rosu Gabriela Nicola
Published: May 2019

e14245

Background: Accurate assessment of tumor response to immunotherapy is challenged by pseudoprogression that mimics true progression. Conventional imaging and RECIST assessment do not adequately distinguish between them given their inability to account for changes in the tumor microenvironment. DPX-Survivac is a novel T cell activating therapy that triggers immune responses against tumors expressing survivin and is being studied in this trial in combination with CPA and pembrolizumab in several solid tumors. Multiparametric MRI approaches - dynamic contrast-enhanced MRI and diffusion-weighted imaging MRI are useful for accurate assessment of structural, perfusion and vascular assessment of the lesion and may identify pseudoprogression and compare to the RECIST-based assessment. Methods: The study will enroll up to 226 evaluable subjects in 5 different cohorts: ovarian cancer, HCC, NSCLC, bladder cancer and MSI-H cancer. These subjects will undergo initial imaging 28 days prior to treatment, to be assessed based on RECIST 1.1, and a pre-treatment tumor biopsy for quantitation of survivin and PD-L1 expression and MSI analyses. Treatment for 35 cycles or until disease progression. All patients will have CT images for RECIST 1.1 and iRECIST assessment. A subset of subjects will undergo mpMRI to calculate advanced imaging biomarkers. Results: MRI, clinical and patient-reported outcomes will be analyzed. Conclusions: This study will provide important evidence on the utility of mpMRI + CT-based assessment of response to immunotherapy and use it as an adjunct to the CT-based RECIST criteria by providing insight on how tumor lesions are impacted by treatment.
Personalized multimodality therapy with immune checkpoint inhibitors (ICI), chemotherapy (CT), and targeted treatment (TT) in advanced refractory cancer.
Authors: Salem Philip A. , Jabboury Khaled W. , Stenoien Randall , Wheeler Thomas M. , Valerio Maria Rosaria
Published: May 2019

e14254

Background: Therapy for advanced refractory malignancies remains a major challenge. As the combination of ICI and CT has been shown in some cancers to be superior to CT alone, and as ICI, CT, and TT have efficacy, but divergent toxicity; this combination was chosen for treating patients with such diseases. Methods: Treatment was highly personalized and was designed based on diagnosis, prior therapy and eligibility for TT. ICI included pembrolizumab in 7 (2 in combination with ipilimumab), nivolumab in 10, and atezolizumab in 2. Chemotherapy was given in 28-day cycles. Multiagent CT cycles were given in 17, and single-agent in 2.13 received platinum and 10 taxanes. TT included erlotinib 4, cituxumab 2, bevacizumab 2, dabrafenib/trametinib 3, crizotinib 1, alectinib 1, sorafenib 1, everolimus 1. 4 were not eligible for TT. Results: From 04/2016 to 10/2018 19 consecutive patients (PT) were treated; median (M), age 62 (27-82), ECOG PS 2 (0-4), female 15/male 4. Tumor type; lung 8, pancreas 3, lymphoma 2, melanoma 2, cholangioCA 1, ureter 1, cervix 1, Glioblastoma Multiforme 1. 13 PT had prior therapy, M2(1-11). 7 had prior ICI, 3 prior TT. Disease was bulky in 6. Response was based on PET evaluation with tumor regression depicted after cycle #1. 11 achieved CR (57%), M 6+ months (1+-15+), 7 PR (37%), M 4 months (1+-7) and 1 SD 7 months. 3 intracranial metastases; 2 CR, 1 PR. Response was seen in extensive prior therapy, bulky disease, poor performance, and prior ICI. No unexpected side effects were observed. Grade 3-4 adverse events; 12.7%, primarily hematologic. Conclusions: The personalized combination of ICI, CT, and TT in advanced refractory diverse cancer diseases is associated with high clinical activity. Further evaluation in clinical trials is strongly indicated.
The role of different TGFβ signatures in predicting outcome in high grade serous ovarian carcinoma.
Authors: Mahdi Haider , Rose Peter Graham , Abdul-Karim Fadi W , Monk Bradley J. , Ni Ying
Published: May 2019

e14262

Background: Ovarian cancer (OV) is the most lethal gynecologic cancer in the US. Despite initial responsiveness to chemotherapy, > 50% experience recurrence. Recurrent disease is characterized by low response to chemotherapy and poor prognosis, especially platinum-resistant disease. Thus, there is an urgent need to identify new targeted therapy to improve the outcome of this aggressive cancer. The Transforming Growth Factor (TGFβ) family controls different cellular responses in development and cell homeostasis. Disruption of TGFβ signaling has been implicated in many cancers, including OV, but with paradox effects. It is unclear if TGFβ family genes can serve as a predictive marker and be selected for future therapy target in OV. Methods: We used quantified RNAseq data from 374 OV samples in The Cancer Genomic Atlas (TCGA) to extract 4 different sets of TGFβ pathway genes including: 3-gene ( TGFB1, TGFB2, TGFB3), 6-gene ( TGFB1, TGFB2, TGFB3, TGFBR1, TGFBR2, TGFBR3), 43-genes from cBioPortal, or 173-genes from Calon et al, 2012 to predict colorectal cancer relapse. Expression counts for these genes were normalized and log transformed to generate a TGFβ score based on the quartiles of the average. Samples were categorized in to low (lowest quartile), medium (middle 2 quartiles), and high (highest quartile) expression groups. Overall (OS) and progression-free (PFS) survivals were compared among groups using Kaplan-Meier method. Results: Interestingly, the minimum TGFβ 6-gene set can predict high grade serous ovarian cancer patient survival among all 4 gene panels using TCGA data, with average age of 60 years (range: 38 –81). Majority of the patients were stage IIIIV. OV patients with highest quartile, TGFβ signature expression have in general less somatic mutations and copy number alterations (average mutation count = 41, average CNA fraction = 0.5) compared to lowest quartile TGFβ expression group (average mutation count = 52, average CNA fraction = 0.6). Patients with TGFB 6-gene expression scored high showed worse OS (36.2 months vs 63.5 months, log-rank p = 0.001) and worse PFS (18.6 months vs 31.4 months, log-rank p = 0.09). Details of outcome by each specified gene signature will be outlined later. Conclusions: Our analysis suggested that the narrowed 6-gene TGFβ family expression profile could potential serve as a prognosis biomarker for OV. With proper validation and mechanistic investigation, existing TGFβ inhibitors may hold promise for targeted or combination therapies for OV. Targeting both TGFβ and PD1 might be of therapeutic value.
Tumor mutational burden assessed by a targeted NGS assay to predict clinical benefit from immune checkpoint inhibitors in non-small cell lung cancer.
Authors: Rothschild Sacha , Alborelli Ilaria , Leonards Katharina , Leuenberger Laura P , Savic Prince Spasenija , Mertz Kirsten D , Poechtrager Severin , Zippelius Alfred , Laubli Heinz Philipp , Haegele Jasmin , Tolnay Markus , Bubendorf Lukas , Quagliata Luca , Jermann Philip
Published: May 2019

e14266

Background: In non-small cell lung cancer (NSCLC) immune checkpoint inhibitors (ICIs) significantly improve overall survival (OS). Tumor mutational burden (TMB) has emerged as a predictive biomarker for patients treated with ICIs. Here we evaluated the predictive power of TMB measured through / by the Oncomine Tumor Mutational Load (TML - Thermo Fisher Scientific) targeted sequencing assay in 71 NSCLC patients treated with ICIs. Methods: TMB was assessed retrospectively in 71 metastatic NSCLC patients receiving ICI therapy. Clinical data (RECIST 1.1) were collected and patients were characterized as either having durable clinical benefit (DCB) or no durable benefit (NDB). Additionally, genetic alterations and PD-L1 expression were assessed and compared with TMB and response rate. Results: TMB was significantly higher in patients with DCB compared to patients with NDB (median TMB = 9.2 versus 5.3 mutations/Mb, Mann-Whitney p = 0.014). 70% of patients with high TMB (cutoff = 3rd tertile, TMB ≥ 9.2) were responders (DCB) compared to 29% of patients with low TMB (cutoff = 1st tertile, TMB ≤ 4.5). TMB-high patients showed significantly longer progression-free survival (PFS) and OS (log rank test, p = .0030 for PFS and 0 .0375 for OS, respectively). Combining PD-L1 expression and TMB value increased the predictive power of TMB. Conclusions: Our results show that the TML panel is an effective tool to stratify patients for ICI treatment. We believe that a combination of biomarkers will maximize the precision of patient selection.
Comparison of biomarkers among 34,855 GI cancer samples show heterogeneity of tumor types.
Authors: Dawood Shaheenah S.
Published: May 2019

e14287

Background: Recent data indicate that biomarker driven use of targeted therapy and I/O-therapy among patients with GI cancer is associated with improved outcome. The presence of biomarkers varies broadly between different GI tumor types, highlighting the importance of comprehensive molecular profiling.To analyze the presence of various alterations in GI cancer samples of a large database, comparing congruency between various tumor types. Methods: A retrospective data analysis of 34855 GI cancer patients profiled at CARIS Life Sciences obtained from Jan 1st2010 till Sep 14th2018 was performed. GI tumors were classified as : CRC, esophageal/gastric/GIST, small intestine, pancreatic/hepatobiliary/liver. Technologies used to analyze the biomarkers: IHC for PD-L1, MMR and Her2, and DNA-NGS for EGFR, BRAF, KRAS, NRAS, MET-CNV, TMB, MSI, POLE and BRCA1/2. Results: Median age was 61 (range 18-89 years). 51.5% was CRC (n = 18047), 15.5 % was esophageal /Gastric/GIST (n = 5470), 3% was small intestinal cancer (n = 886) and 30% was pancreatic/hepatobiliary/liver cancer (n = 10452). Information on biomarkers was available from 2931 cases for MET amplification to 28536 for RAS information. Overall, the most common finding was a pathogenic RAS-mutation (either KRAS or NRAS) in 7650 cases (26.81%), the rarest one was a mutation in EGFR in 12 cases (0.06%). Higher rate of HER2 amplification was observed among pts with esophageal/gastric/GIST tumors (6.5%) in comparison to tumors at other GI sites ( ~1.5%). High TMB was seen among patient with CRC and small intestine tumors (~7%) while it was lowest among pancreatic cancer (1.8%). Compared to other GI sites lower MSI /MMR deficiency rates were observed in pancreatic /hepatobiliary tumors, significantly higher PD-L1 positivity was observed in gastrooesophageal cancer types, increased MET-amplifications in gastrooesophageal and small intestinal cancer types and lower RAS-mutation rate in gastrooesophageal cancer. Conclusions: Molecular profiling analyzing druggable biomarkers can help identify patients with increased likelihood for benefit from immune-checkpoint-inhibitors and targeted therapies. Further investigations are needed to evaluate the different findings in various GI cancer types.
Arenavirus-based vector platform for massive tumor self-antigen-specific CD8 T cell immunity.
Authors: Orlinger Klaus Karl , Bonilla Weldy V. , Kallert Sandra M , Kirchhammer Nicole , Marx Anna-Friederike , Krzyzaniak Magdalena , Schmidt Sarah , Raguz Josipa , Berka Ursula , Guenther Stephan , Zippelius Alfred , Pinschewer Daniel
Published: May 2019

e14297

Background: The induction of powerful CD8+ T cell immunity to tumor associated self-antigens (TAAs) represents a critical yet challenging goal. Here we report on the development of an arenavirus-based delivery platform meeting this challenge. Previously we have shown that genetically engineered replication-attenuated lymphocytic choriomeningitis virus (LCMV) vectors, TheraT(LCMV), induce strong TAA-specific CD8 T cell immunity, but these responses can not be substantially augmented upon TheraT(LCMV) readministration. Counter to expectations, vector-neutralizing antibodies were not accountable for limited homologous prime-boosting capacity. Instead, dominant viral backbone-reactive CD8+ T cells competed against subdominant TAA-specific responses, limiting their magnitude. Methods: Herein we engineered and characterized delivery systems based on the arenaviruses Mopeia, Candid#1 and Pichinde (TheraT(MOP), TheraT(CAND), TheraT(PIC)). Results: We demonstrate that heterologous TheraT(CAND) – TheraT(LCMV) and TheraT(PIC)-TheraT(LCMV) prime-boost substantially augment TAA-specific CD8 T cell responses by rendering them immunodominant. Accordingly, intravenous administration of mice triggered up to 50% TAA epitope-specific CD8+ T cells and cured established tumors. Conversely, TheraT(MOP) – TheraT(LCMV) prime-boost was poorly immunogenic owing to cross-reactive T cell epitopes in the respective viral backbones. Conclusions: These findings establish heterologous arenavirus prime-boost combinations as a powerful new modality in tumor immunotherapy and highlight CD8 T cell epitope dominance as a significant hurdle to overcome in the vectored delivery of TAAs.
Enhanced CD138 peptide-specific cytotoxic T lymphocyte activities against breast, colon and pancreatic cancers in combination with pembrolizumab (anti-PD1).
Authors: Bae Jooeun , Hideshima Teru , Tai Yu-Tzu , Munshi Nikhil C. , Anderson Kenneth Carl
Published: May 2019

e14302

Background: We investigated the ability of CD138 antigen-specific CD8+ cytotoxic T lymphocytes (CD138-CTL), generated using a novel HLA-A2-specific CD138260-268 (GLVGLIFAV) peptide, in combination with various immune checkpoint inhibitors, to target breast, colon, and pancreatic cancer. Methods: This study was designed to evaluate anti-tumor activities of CD138-CTL, alone or in combination with Pembrolizumab (anti-PD1), against solid tumors. Results: Upregulation of CD138 protein expression was detected on breast, colon, and pancreatic cancer cells. CD138-CTL induced ex vivo by repeated stimulations with HLA-A2-specific CD138260-268 (GLVGLIFAV) peptide were enriched for CD45RO+ memory CTL and demonstrated antigen-specific and HLA-A2-restricted anti-tumor activities. Treatment of CD138-CTL with anti-PD1, anti-TIM3, or anti-LAG3 or tumor cells with anti-PDL1, anti-Gal9 or anti-LAG3 resulted in an increased proliferation of CD138-specific CD8+ CTL, which express costimulatory and activation markers (CD28, CD40L, 41BB, CD69, CD38). However, expression of immune checkpoints (PD1, LAG3, CTLA4) was also induced on these CD138-CTL upon stimulation. Importantly, anti-tumor activities of CD138 peptide-CTL was significantly enhanced in combination with Pembrolizumab, especially within the antigen-specific memory CD8+ T cell subsets (central memory > effector memory), directed against HLA-A2+ breast cancer, colon cancer and pancreatic cancer. Conclusions: These data provide the framework for an immunotherapeutic strategy encompassing a CD138260-268 (GLVGLIFAV) peptide-based cancer vaccine in combination with Pembrolizumab to enhance antigen-specific central memory cells with more potent and long-lasting CD8+ CTL immune responses in patients with solid tumors. Based on these result, clinical trials evaluating CD138260-268 (GLVGLIFAV) in multi-peptide cancer vaccine are on-going in patients with triple negative breast cancer in combination with anti-PD1 (Pembrolizumab) in metastatic setting (NCT 03362060) or anti-PD-L1 (Durvalumab) in stage II/III, adjuvant setting (NCT 02826434).
Correlation betwen MKK4 expression (exp) and clinicopathological features, KRAS/NRAS mutation in metastatic colorectal cancer (mCRC).
Authors: Dogan Mutlu , Guresci Servet , Kos Fahriye Tugba , Bal Oznur , Ergun Yakup , Bozdogan Onder
Published: May 2019

e14500

Background: We aimed to evaluate correlation between MKK4 exp & clinicopathological features in mCRC Methods: MKK4 exp was assessed by immunoreactive score (IRS). Staining intensity [0 (no), 1+ (weak), 2+ (moderate), 3+ (strong)] & % of positively stained cells [0 (0%), 1( < 10%), 2 (11-50%), 3 (51-80%), 4 ( > 80%)]. IRS was calculated by this formula (IRS = SI × PP). Cut-offs were explored with ROC analysis. AUC was 0.629 (95%CI:0.419-0.840) & best cut-off value for MKK4 was 2.5 (sensitivity of 68 %). Groups were WIR ('weak'; IRS:0-2) & SIR ('strong'; IRS ≥3). Results: We enrolled 95 patients. Median follow-up was 31.4 months. MKK4 exp strength outcomes are shown in table. K-ras/N-ras mutation rate was 45.2%. WIR patients had longer OS (p: 0.03). In WIR group, metastatic ones at diagnosis had shorter DFS (p:0,05). There was no PFS difference (p: 0,15). In correlation analysis, there was a a negative correlation between MKK4 expression & 1st line treatment resistance, k-ras/n-ras mt, OS, PFS, DFS (r: -0.01; r: -0,06; -0,02; -0,10; -0,06; -0,34) ( p:0,04 for DFS). Conclusions: MKK4 exp inversely correlates with survival outcomes & ras mutation. WIR leads longer DFS & OS. [Table: see text]
High dose apatinib in the reversal of immunosuppressive tumor microenvironment for irradiation therapy in lung carcinoma.
Authors: Liang Lijun , Hu Chenxi , Wen Yixuan , Hui Kaiyuan , Jiang Xiaodong
Published: May 2019

e14503

Background: The purpose of this study was to investigate the potential systemic anti-tumor effects of stereotactic ablative radiotherapy (SABR) and apatinib (a novel vascular endothelial growth factor receptor 2 tyrosine kinase inhibitor) via reversing the immunosuppressive tumor microenvironment for lung carcinoma. Methods: Lewis lung cancer cells were injected into C57BL/6 mice in the right hindlimb (primary tumor; irradiated) and in the left flank (secondary tumor; nonirradiated). When both tumors grew to palpable size, mice were randomly divided into eight treatment groups. These groups received normal saline or three distinct doses of apatinib (50 mg/kg, 100 mg/kg, 200 mg/kg) daily for 7 days, in combination with a single dose of 15 Gy radiotherapy to the primary tumor or no radiotherapy. Further tumor growth/regression of mice was followed and observed. Results: For the single 15 Gy modality, delays in tumor growth could only be observed at the primary tumor. When combining SABR and apatinib 200 mg/kg, significant retardation of both primary and secondary tumor growths could be observed, indicating that an abscopal effect had been induced. Mechanism analysis suggested that programmed death ligand-1 expression increased with SABR and was counteracted by additional apatinib therapy. Furthermore, when apatinib was combined with SABR, the composition of immune cells could be changed. More importantly, this two-pronged approach evoked tumor-antigen-specific immune responses, and the mice were resistant to subsequent tumor rechallenge. Ultimately, long-term survival was improved. Conclusions: Our results suggested that the tumor microenvironment could be managed with high-dose of apatinib, which was effective in eliciting an abscopal effect induced by SABR.
Clinical assessment of tumor angiogenesis and its predictive value for chemotherapy responses of colorectal metastatic hepatic tumors.
Authors: Okamoto Hirotaka , Iino Hiroshi , Shiba Shugo , Sudoh Makoto , Maruyama Suguru , Wakana Hiroyuki , Ichikawa Daisuke , Fujii Hideki
Published: May 2019

e14505

Background: The treatment of colorectal cancer (CRC) with metastasis is now widely accepted to be systemic chemotherapy of 5-FU base chemotherapy (FOLFOX or FOLFILI) in combination with molecular antibody. The objective of this study was to determine the significance of tumor angiogenesis in metastatic hepatic tumors in relation to the response of systemic chemotherapy. Methods: A total of 48 consecutive CRC patients with hepatic metastasis who had been treated with or without hepatic resection were retrospectively reviewed, and factors, such as metastatic tumor angiogenesis and chemotherapy response, were analyzed. Tumor angiogenesis in association with ring-enhancement (RE) on computed tomography (CT) was also microscopically evaluated by microvessel counting (MVD) in sections stained immunochemically with CD34. Results: The overall response rate after 6 courses of first-line chemotherapy for the liver metastasis with RE on CT was 64% (23/36), whereas the response rate without RE was 25% (3/12), which was significantly different. The survival rate of patients with or without liver resection of all RE-positive tumors was not different, but the survival rate of patients with liver resection was longer than that of patients without liver resection for all RE-negative tumors. Microscopic examination revealed that peripheral RE on CT of the metastatic tumor was associated with tumor angiogenesis by MVD. Conclusions: The peripheral RE on CT of the metastatic hepatic tumor was associated with tumor angiogenesis and may predict chemotherapy response. A combination of liver resection with chemotherapy improves the survival of patients who have multiple hepatic metastasis.
Tracking endocrine resistance in estrogen-receptor positive breast cancer in ctDNA.
Authors: Meissner Magdalena , Butler Rachel , Casbard Angela Claire , Carucci Margherita , Madden Tracie-Ann , Howell Sacha Jon , Jones Robert Hugh
Published: May 2019

e14550

Background: Endocrine therapy is the standard of care treatment for patients with estrogen-receptor positive advanced breast cancer, owing to improved tolerability and comparable efficacy to that of cytotoxic chemotherapy. Half of such cancers will progress through first line therapy (primary endocrine resistance) and half will progress after an initial period of disease control (secondary or acquired endocrine resistance). A significant challenge is to test for and identify biomarkers which can guide the likely success of endocrine therapy as a single agent or in combination with small molecule inhibitors.This is particularly challenging where metastatic deposits reside at sites where biopsy is difficult. Potential biomarkers indicative of resistance to endocrine therapy have been identified and can be detected in circulating tumour DNA (ctDNA). CtDNA is shed from tumours and is detectable in a cancer patient’s bloodstream. Information on mutational profiles can guide an oncologist in the selection of targeted therapy in addition to hormonal therapy. Methods: We have analysed formalin-fixed paraffin-embedded(FFPE) tumour samples and longitudinal liquid biopsies from 19 patients who were treated with fulvestrant in combination with a novel inhibitor of the PIK3CA/AKT pathway with next-generation sequencing using a targeted 44-gene panel. Mutations identified using this technique were tracked during the course of treatment using droplet-digital PCR(ddPCR). Results: 57 samples were tested using a 44-gene panel; 19 FFPE tumour samples and matched ctDNA samples were obtained prior therapy and ctDNA samples at disease progression. Mutations detected in PIK3CA, AKT1, ESR1and TP53 genes were trackable in longitudinal ctDNA samples using ddPCR. The association between ctDNA dynamics and outcome will be presented. Conclusions: Multiple mutations that enable the early detection of treatment failure and resistance can be tracked in ctDNA. Investigating the clinical utility of ctDNA is paramount.
Prediction of lesion shrinkage using CT imaging with radiomic and deep learning approaches.
Authors: Ma Junshui , Liu Rongjie , Goldmacher Gregory V. , Baumgartner Richard
Published: May 2019

e14592

Background: Radiomic features derived from CT scans have shown promise in predicting treatment response (Sun et al 2018, and others). We carried out a proof-of-concept study to investigate the use of CT images to predict lesion-level response. Methods: CT images from Merck studies KEYNOTE-010 (NCT01905657) and KEYNOTE-024 (NCT02142738), were used. Data from each study were evaluated separately and split for training (80%) and validation (20%) in each study. A lesion was classified as “shrinking” if ≥30% size reduction from baseline was seen on any future scan. There were 2004 (613 shrinking vs. 1391 non-shrinking) and 588 (311 vs. 277) lesions in KN10 and KN24, respectively. 130 radiomic features were extracted, followed by random forest to predict lesion response. In addition, end-to-end deep learning was used, which predicts the response directly from ROIs of CT images. Models were trained in two ways: (1) using pre-treatment baseline (BL) only or (2) using both BL and the first post-treatment image (V1) as predictors. Finally, to evaluate the predictive power without relying on initial lesion size, size information was omitted from CT images. Results: Results from the KN10 and KN24 are summarized in Table. Conclusions: The results suggest that the BL CT images alone have little power to predict lesion response, while BL and the first post-baseline image exhibit high predictive power. Although a substantial part of the predictive power can be attributed to change in ROI size, the predictive power does exist in other aspects of CT images. Overall, the radiomic signature followed by random forest produced predictions similar to, if not better than, the deep learning approach. [Table: see text]
A preliminary single institution experience with droplet digital polymerase chain reaction (dd-PCR) liquid biopsy (LB) for therapeutic decision in advanced solid tumors: Personal precision oncology clinic and genetic diagnostics, Brazil.
Authors: Murad Andre Marcio , Rocha Jose Claudio Casali
Published: May 2019

e14613

Background: Droplet digital polymerase chain reaction (dd-PCR) is a promising method for analyzing minor amounts of cell-free circulating free nucleic acid (DNA and RNA) due to its high sensitivity, low cost, and fast reading, since it dispenses bioinformatics, making it an appropriate alternative to new generation sequencing (NGS) for the detection of biomarkers to guide molecularly targeted cancer therapy. The assay covers main actionable hotspot alterations across many actionable genes: EGFR(mutations), ALK (fusion, mutations), ROS1(fusion), BRAF(mutations), KRAS (mutations), NRAS(mutations), ERBB2(CNV), ESR1(mutations), KIT(CNV) and PDGFRA(CNV). Methods: We analyzed so far 38 patients with advanced NSCLC: 13(35%), breast: 9(24%), colorectal: 6(16%), pancreatic: 4(10%), ovary: 2(5%), salivary gland: 2(5%), melanoma 1(2.5%), and GIST: 1 (2.5%). ddPCR was performed using the QX200 system (BIO-RAD, Hercules). All samples were tested in duplicate. All assays were performed at Personal Precision Genetic Diagnostics. Belo Horizonte, MG. Brazil. Results: We detected meaningful genomic alterations in 7(18.5%) patients: 4(10.5%) KRAS G12V mutations (all in colorectal cancer), 1(2.6%) EGFR Del19 mutation, 1 EGFR L858R mutation(2.6%), both in NSCLC and 1(2.6%) ERBB2 amplification (in breast cancer). MAF (Mutant Allele Fraction) varied from 0.9% to 24%. In all cases the results were decisive for the indication or the change of a targeted therapy. Conclusions: Our preliminary results suggest that dd-PCR is a highly sensitive method and could be used for a routine laboratory detection of the important genomic variations to determine the targeted therapy in patients with varied advanced solid tumors.
Oncology precision medicine for hepatobiliary and pancreatic cancer: An institutional review.
Authors: Bellini Geoffrey , Godden Jennifer Jo , Weese James L. , Chevinsky Aaron , Papenfuss Wesley Allan , Thompson Michael A.
Published: May 2019

e14626

Background: Hepatobiliary cancers - hepatocellular carcinoma (HCC), intra or extrahepatic cholangiocarcinoma (I/EC), and gallbladder carcinoma (GB) - and pancreatic adenocarcinoma (PC) remain a leading cause of death with little improvement in long-term outcome. Recent studies have suggested that these cancers harbor actionable mutations to varying degrees. The aim of our study was to examine the number of patients (Pts) with these primary tumors who underwent molecular testing in a large vertically integrated health system. Subsequently, we analyzed the percentage of that population who may be candidates for oncology precision medicine (OPM) directed therapy. Methods: We identified Pts with HCC, IC, EC, GB in an IRB reviewed OPM database of our system over a one year period. Pts who underwent molecular panel testing were selected out, and their molecular alterations were identified and stratified by cancer type. Results: 304 total Pts were identified. 61 (20%) underwent molecular testing broken down as follows: 17/132 (13%) I/EC and HCC, 3/11 (27%) GB, and 41/161 (25%) PC. Quantity not sufficient for testing was in 10/61 (16%), of which 5/10 (50%) were resubmitted and tested successfully. 6/61 (10%) were cancelled or deemed not appropriate. Test recommended potential actionability was 8/17 (47%) of I/EC and HCC, 2/3 (67%) of GB, and 25/41 (61%) of PC. Conclusions: OPM is a dynamic area of increasing testing and learning. We found 13-27% of hepatobiliary and pancreatic Pts had molecular testing, which suggests the potential to increase molecular screening for this difficult group of tumors. Total genetic alterations (TGA) and clinically relevant genomic alterations (CRGA) per patient are similar to Ross et al. ( http://ow.ly/k52a30nBMnU ) for GB. Final interpretation regarding pragmatic actionability (patient on drug) and clinical outcomes are still under investigation.[Table: see text]
A novel mutant-allele capture method for enrichment of low abundant cancer mutants.
Authors: Lin Rui , Pu Yue , Mao Li
Published: May 2019

e14632

Background: High sensitivity detection of cancer mutants can facilitate cancer diagnosis, treatment selection, and disease monitoring. Improved methods to detect low abundance of cancer driver mutants are needed. Methods: We developed a novel method, termed PEAC, with a super high sensitivity to detect low abundant mutant from cancer driver genes through mutant capturing followed by Sanger sequencing. Circulating free DNA (cfDNA) standards containing 20 ng of 5%, 1%, 0% (wild-type), or 40 ng of 0.1% cancer driver mutants were used to establish the method. After pre-amplification of both wild-type and mutant alleles, mutant fragments were specifically bound by biotin-labelled, locked–nucleic acid modified probes, enriched by streptavidin beads, amplified through PCR and sequenced using Sanger for detection. Results: The wild-type standard did not show any mutation signal either with or without PEAC, nor did the standards with 5%, 1%, or 0.1% mutants prior to PEAC enrichment. However, after PEAC enrichment, 5%, 1%, and 0.1% of cancer driver mutants ( EGFR L858R, EGFR T790M, and BRAF V600E) predicting sensitivities to target therapies were enriched to ~90%, ~80% and 30-40% abundance, respectively. In addition to the application for point mutations, PEAC methodology was also used to enrich deletion mutant; 5%, 1%, and 0.1% of EGFR exon 19 deletion, E746_A750del, were all enriched to 90-100% frequency after PEAC. To address the limit of detection of PEAC technology, 100 ng of cfDNA standard containing 0.01% EGFR L858R and EGFR exon 19 deletion mutant were enriched to 15% and 50%, respectively, after PEAC, which indicates up to 5000-fold enrichment for EGFR exon 19 deletion by PEAC technology. Conclusions: PEAC technology can enrich very low abundance of key cancer driver mutants predicting sensitivities to tyrosine kinase inhibitors. PEAC enrichment in combination with Sanger sequencing may guide the application of cancer target therapy and represent a useful technology in the era of precision medicine.
The chick embryo chorioallantoic membrane assay: In ovo model for personalized assessment and evaluation of the most effective therapeutic approach in cancer therapy.
Authors: Shapira Shiran , Bogin Oren , Kazanov Dina , Bedny Faina , Hershkovizh Dov , Izhaki Ayelet , Brazowski Eli , Bernstein Ezra , Wolf Ido , Arber Nadir
Published: May 2019

e14634

Background: "Personalized medicine,” is the tailoring of medical treatment to a single person aiming to maximize efficacy and minimize toxicity. Currently there is no good prediction for response to therapy. The Chick Chorioallantoic Membrane (CAM) is naturally immuno-deficient and rich in vascularity therefore an ideal system, allowing generate 3D cancerous “organoids” in a very efficient, reproducible and cost-effective manner and translates basic research to the clinic. Aims: Generate a “personalized” HTP system for a quick, reliable and effective evaluation of different therapeutic options using 3D tumors in a "humanized egg" instead of mouse PDX model. Methods: Fertilized eggs were incubated until day 3 (37°C, 75-90% humidity). Then, 2ml of albumin was pulled out to separate the CAM from the eggshell and a small window in the eggshell has been made. To destroy the chicken immune system development, the eggs were irradiated at day 5 and human immune cells were then inoculated onto the CAM. On day 7, single cells suspension or tissues, derived from cancer patients, were transplanted onto the CAM and visible tumors were performed ("CAM-PDX"). Drugs, mAbs and chemotherapy, were applied via the yolk sac. Tumor growth was measured, weighted and monitored by caliper and IVIS fluorescent imaging platform. IHC was performed and confirmed the response of the particular specimens to the tested regiment. Results: Histology and IHC analysis confirmed that the established tumors retained their characteristics. Positive Ki-67 staining confirmed that cancer cells proliferate while the treated tumors showed reduced staining. Anti-CD24 mAb, FOLFOX, cetuximab, Foflorinox and Gemcitabine, given as single agent or combinations, successfully inhibited CR and pancreatic tumors (by 70-75%). Detection of active caspase 3 confirmed those results. Biopsies from human specimens, were successfully established and expanded by serial passages allows generation of bio-bank. The stimulated human PBMCs demonstrated enhanced proliferation in vitro and in ovo, even after 5 days in the egg. Irradiated eggs showed no functional immune system even after 2 weeks of development. Conclusions: The CAM is an ideal, effective, economical and powerful avatar-based precision medicine approach to predict the best protocol for cancer therapy.
Non-inferiority of a new advanced dry plasma thawing system compared to leading standard method: Preservation of coagulation and anticoagulation proteins necessary for use in the clinic.
Authors: Bischoff Farideh Z. , Davey Will , Read Brian , Deleon Tess , Laba Rebequec , Thacher Fred , Shavit Moni
Published: May 2019

e14675

Background: Cancer can affect the bone marrow, cause anemia and internal or excess bleeding, requiring blood or plasma transfusion. Fresh frozen plasma (FFP) should be thawed before transfusing to patients but it must be monitored closely to avoid contamination, uncontrolled or prolonged thawing which may subsequently lead to plasma protein denaturation. In this study, we compared and evaluated the effect of two different thawing methods on clotting factor activities as well as factors such as throughput and turnaround time. Methods: Over a period of approximately one month, November 6 to December 3, 2018, our testing laboratory at the UCSD School of Medicine received three sets of samples for a total of 197 freshly collected (pre-freeze) or freshly thawed plasma samples (standard commercial thawing device and the new dry ZipThaw device) that had been collected and processed in accordance with a San Diego Blood Bank protocol described elsewhere. The Lab conducted over 1,782 tests on these plasma for the following factors: Prothrombin time (PT), International Normalized Ratios (INR), Activated Partial Thromboplastin time (aPTT) Factor VIII activity, Factor V activity, Protein C activity, Protein S antigen, Von Willebrand activity and Thrombin Antithrombin Complex (TAT) concentration. The assays were carried out on an ACL TOP 700 Hemostasis Analyzer (Instrumentation Laboratory, Bedford, MA, USA) using manufacturer’s reagents and automated protocols. The ACL TOP 700 has validated precision CV of 5- 10%. All assays were completed within 4 hours of collection or, if frozen, within 4 hours of thawing. The statistical analysis method employed is two-tailed, student t-test. Results: We observed 100% concordance with no statistically significant differences between the pre-freeze group and the two different post thaw groups, except for a slight increase in the mean TAT complex concentration. Overall, the means of all three groups were well within the normal range defined by the assay kits (1-10 ng/ml). Further, the increased mean TAT is more notable with standard method (33%, p = 0.004) than with the ZipThaw (18%, p = 0.054). Conclusions: We report for the first time non-inferiority between the new ZipThaw device and the leading commercial predicate in plasma thawing while preserving coagulation and anticoagulation proteins and minimizing activation of clotting cascade. The results provide support for the use of ZipThaw device in clinical settings.