bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of hydrogen gas on exhausted CD8+ T cells (PD-1-positive, terminally differentiated CD8+ T cells).
Authors: Akagi Junji , Baba Hideo
Published: September 2018

e23071

Background: CD8+ T cells progress from an early-differentiated (early-CD8+ T cells) to a terminally differentiated state (terminal-CD8+ T cells). PD-1 expression is rapidly upregulated in antigen-activated T cells, and rapidly downregulated when the antigen is cleared. In the presence of persistent antigen stimulation, such as that in cancer patients, PD-1 expression is not downregulated and terminal-CD8+T cells become exhausted. Methods: Using flow cytometry, we investigated PD-1 expression in CD8+T cells in the peripheral blood of 37 patients with stage IV cancer, before and after they received hydrogen gas treatment. We analyzed associations between treatment and progression-free survival (PFS) using Cox and Kaplan-Meier survival analyses. Results: Treatment lead to partial response (PR) in 12 patients and stable disease (SD) in 14 patients, with response rates of 32.4% and 37.8%, respectively (with an overall clinical response rate of 70.3%), and reduced the number of PD-1-positive, terminal-CD8+ T cells in 9 of 12 patients with PR (75%) and in 8 of 14 patients with SD (57%). However, in 8 of 11 patients with progressive disease (PD) (72.7%), these cell counts increased. Multivariate analyses demonstrated that higher PD-1-positive, terminal-CD8+ T cell counts were associated with shorter PFS periods, both before (hazard ratio [HR] = 3.23; 95% confidence interval [CI], 1.475–7.070; p = 0.003) and after (HR = 2.75; 95% CI, 1.273–5.951; p = 0.01) treatment, although the hazard ratio was decreased by treatment. Additional analysis showed that patients with higher PD-1-positive, terminal-CD8+ T cell counts had shorter survival rates than those with lower counts, both before and after treatment (log rank test, p = 0.001 and p = 0.005, respectively). The 20-month survival rates for patients with higher PD-1-positive, terminal-CD8+T cell counts improved from 0% to 36.7% after treatment. Conclusions: These results suggest that PD-1-positive, terminal-CD8+ T cell counts are highly associated with PFS duration in patients with stage IV carcinomas, and hydrogen gas treatment contributes to clinical outcomes by reducing PD-1-positive, terminal-CD8+ T cell counts.
Role of the inflammasome of tumor cells in modulating the biology of Tumor Infiltrating T lymphocytes (TILs) in colorectal cancer.
Authors: Jarry Anne , Ouairy Adrien , Dansette Delphine , Deleine Cécile , Jouand Nicolas , Oger Romain , Podevin Juliette , Fourquier Pierre , Thibaudeau Emilie , Chetritt Jerome , Mosnier Jean-François , Gervois Nadine , Bossard Celine
Published: September 2018

e23087

Background: In colorectal cancer (CRC), little is known about mechanisms by which tumor cells can influence the phenotype and biology of Tumor Infiltrating T lymphocytes (TILs) of the tumor microenvironment. One of these mechanisms could be the inflammasome, a molecular platform present in normal intestinal epithelial cells, whose effector protein, caspase-1, can rapidly mature IL18 and generate a mucosal Th1/Tc1 (IFNγ) response. However, the inflammasome status of tumor cells in CRC and its potential role on TILs are unknown yet. Methods: Prospective and retrospective cohort studies aimed to determine in CRC patients: the status of the inflammasome in tumor cells (IL18 immunostaining on tissue microarrays (TMA) and in situ detection of active caspase-1 on frozen sections) and the density of TILs (CD8+, T-bet+) in relation with i) the microsatellite stable (MSS) or unstable (MSI) status of CRC, and ii) the levels of cytokines (IL18, IFNγ) secreted in an ex vivo explant culture model of CRC. Finally, we assessed the effect of recombinant human IL18 (rhIL18) on the IFNγ response of isolated TILs. Results: TMA analysis of the retrospective cohort showed that IL18 was significantly expressed (in more than 50% of tumor cells) in 80% of CRC, especially in MSI CRC, and correlated with a high density of T-bet+ and CD8+ intraepithelial TILs (IEL-TILs). Active caspase-1 was detected in tumor cells in 60% of CRC. In the prospective cohort, the presence of active caspase-1 in tumor cells was associated with high levels of mature IL18 secreted in explant cultures, with high density of T-bet+ TILs and with IFNγ release in most cases. In addition, isolated TILs expressing IL18 receptors (IL18Rα), cultured with rhIL18, were able to secrete IFNγ either unstimulated or stimulated with OKT3. Conclusions: The inflammasome of tumor cells, when maintained and active, can contribute to a Th1/Tc1 antitumor immune response elicited by TILs, that can modulate tumor growth. The inflammasome of tumor cells can thus be considered as a potential new therapeutic target to strengthen the antitumor immune response in CRC, in association with other immunotherapies.
A new method for gene expression analysis of pure lymphocytes recovered from heterogeneous fixed mouse tissue.
Authors: Chow Jennifer , Da Silva Ana Paula Galvão , Medoro Gianni , Manaresi Nicolò , Lira Paul David , Bischoff Farideh Z.
Published: September 2018

e23089

Background: Tumor infiltrating lymphocytes (TILs) are biomarkers that play a critical role in cancer diseases, including differential diagnosis, determination of prognosis, prediction of response to treatment, and evaluation of disease progression. Gene expression analysis in TILs derived from fresh tissue may not accurately depict the gene profile of the tissue microenvironment as it can change aggressively during lymphocyte isolation and RNA extraction. In addition, tissue sample size can limit the isolation of TILs with current technologies. In this study, we demonstrate the use of the DEPArray™platform to isolate pure populations of lymphocytes from a fixed mouse tissue for RNA analysi. Methods: Mouse splenocytes were activated in vitro with anti-CD3 and -CD28 for 72hs. Cells were harvested, fixed with 2% paraformaldehyde (PFA) for 20 min at RT, and stained for either CD4 or CD8 expression. Gene expression analysis of CD45, ADORA2A, GLS and GAPDH was performed in CD4+ and CD8+ DEPArray™sorted cells using the TaqMan PreAmp Cells-to-Ct kit. Results: The table below summarizes the Ct values for CD45, ADORA2A, GLS and GAPDH expression in 300 fixed unsorted control and DEPArray™sorted lymphocytes. Conclusions: We have demonstrated the feasibility of gene expression analysis on pure populations of CD4+ and CD8+ cells isolated from a fixed tissue using the DEPArray™ platform. The advantage of this approach is the DEPArray’s ability to identify and isolate subpopulations of cells from complex heterogeneous samples and/or specimens that are limited by size or content. This methodology will be applied for isolation of TILs in syngeneic and xenograft models of cancers for downstream RNA applications. [Table: see text]
The role of diffusion weighted magnetic resonance imaging for the characterisation of mediastinal lymphadenopathy.
Authors: Garg Mandeep K , Ramamoorthy Eniyavel , Singh Paramjeet , Aggarwal Ashutosh Nath , Gupta Nalini
Published: September 2018

e23113

Background: To assess the diagnostic performance and additional value of diffusion-weighted (DW) magnetic resonance imaging (MRI) in the characterization of mediastinal lymphadenopathy. Methods: This prospective study was approved by the institutional review board, and written informed consent was obtained from all patients. A total of 43 patients [23 males and 20 females with mean age of 49 years] with mediastinal lymphadenopathy underwent DW and T2 weighted MRI and pathological examination in the period of January 2015 to June 2016. Diffusion restriction pattern, apparent diffusion coefficient (ADC) mean value, size and T2 heterogeneous signal intensity of the nodes were evaluated. Receiver operating characteristic curve analysis was used to determine ADC mean threshold values. The best predicting combinations of these four parameters were selected by means of stepwise logistic regression analysis. Results: ADC mean value of malignant lymphadenopathy (0.873±0.109×10−3 mm2/s) was significantly different from that of benign lymphadenopathy (1.663±0.311×10−3 mm2/s) (P < 0.001). When an ADC mean value of 1.0955×10−3 mm2/s was used as a threshold value for differentiating malignant from benign nodes, the best results were obtained with a sensitivity of 94%, a specificity of 96% and area under the curve of 0.996. The three other predictor variables didn’t add anything statistically significant to the regression equation with ADC mean. Conclusions: ADC mean value of malignant mediastinal lymphadenopathy was significantly different from the benign lymphadenopathy. The ADC mean value was the strongest independent predictor of malignancy.
Genome-wide microRNA expression profiling in malignant pleural effusion to identify a ten-microRNA signature.
Authors: Zhang Meiling , Zhu Miaojun , Hu Qin , Li Chun , Zhu Zhenhua , Hou Yingyong , Xu Jing , Wang Hailong , Zhu Tong , Tang Tom , Tan Ruoying , Zhang Xin
Published: September 2018

e23123

Background: Pleural carcinosis caused by tumors of the chest (e.g., lung and breast cancer) or elsewhere in the body (e.g., ovarian carcinoma) that metastasize to the visceral and/or parietal pleura. Recurrent malignant pleural effusion due to pleural carcinosis is one of the most common findings in oncology. The aim of the study was to identify a miRNA signature associated with clinicians’ prescription of patients with MPE. Methods: We used qRT-PCR assay to evaluate a wide range miRNAs (1920 miRNAs) signature measured from pleural effusion between patients with cancer and healthy controls. From the data of our previous studies we selected a panel of 96 miRNAs related to malignant pleural effusion. Data were compared by three classification methods software for statistics and modeling. We used a first set of 77 pleural effusion samples as training group, including 27 patients affected by Lung Adenocarcinomas (LAC), 9 with other lung cancer, 17 with other tumors and 24 inflammatory patients as negative controls. Moreover, we used more 64 pleural effusion samples for double-blind predictive study. Data analysis was performed using a machine learning approach of a Support Vector Machine classifier with a Student's t-test feature selection procedure. Results: We identified a panel of ten miRNAs with optimum classification performance. The combination of these 10-miRNAs alone could discriminate MPE cases from negative controls with an AUC of 0.969 (accuracy = 93.5%; specificity = 91.7%). The selected panel of another 10-miRNAs could separate lung cancer cases from negative controls with an AUC of 0.973 (accuracy = 94.8%; specificity = 95.8%), and a small panel of 4-miRNAs could good discriminate LAC cases from negative controls with an AUC of 0.946 (accuracy 88.9%; specificity = 100%). The accuracy rate of the double-blind predictive sensitivity value was 90.9% and specificity value was 95.8% for MPE patients with lung cancer of miRNA signature. Conclusions: Our miRNAs profile may serve as a new biomarker for MPE diagnosis. Otherwise, we identified a 10-miRNAs signature for MPE patients with lung cancer and a 4-miRNAs for MPE patients with lung adenocarcinoma.
Genomic profiling of early-stage lung cancer for patterns of recurrence.
Authors: DiPerna Costanzo A , Fine Leah
Published: September 2018

e23126

Background: Currently there is no evidence to support molecular profiling for early stage resected lung cancer patients. However, up to 50% of patients experience recurrence following resection. This is a first-of-a-kind study utilizing comprehensive genomic profiling technology to characterize genomic patterns for risk of recurrence in early stage lung cancer patients within the community hospital setting who have undergone lung surgery. Methods: A total of 60 Stage I-II lung cancer patients, all having undergone pulmonary resection, were evaluated with molecular profiling of their primary lung cancer tissue using Foundation Medicine’s FoundationOneÒ test. Patient age in years ranged from 39-86, and all patients were confirmed Stage I or II based on final pathologic analysis. Samples were taken from three community hospitals that are part of the Addario Lung Cancer Foundation Centers of Excellence program. Patients whose tumors were resected between the years of 2009-2017 were included in this combined retrospective and prospective study. Results: More than 300 genes were evaluated using FoundationOneÒ and patients were segmented to establish similarities and differences. Analysis of segments include gender, recurrence, smoking status among others. Gene patterns across segments are beginning to reveal possible predictive profiles. Final analysis will be completed shortly. Conclusions: Genomic profiling could help predict lung cancer recurrence for early stage lung cancer patients. Similarities amongst patients with recurrences imply that early genomic profiling of lung cancer patients could help predict those patients who would benefit from adjuvant therapies including conventional chemotherapy. Genomic profiling for early stage lung cancer should be studied further and in greater detail to help predict those patients who would benefit from potentially early adjuvant therapies.
Distribution of ETV6-NTRK3 translocations across neoplasms identified from the Mitelman Database.
Authors: Malpartida Juan Carlos , Vick Eric , Richardson Noah Hunter , Patel Kruti , Stein Matthew K , Martin Michael Gary
Published: September 2018

e23141

Background: Discovered as a novel aberration in congenital fibrosarcoma (CF), the ETV6-NTRK3 translocation (EN) confers oncogenic potential and is inhibited by crizotinib. The present study aims to survey the scope of neoplasms that harbor EN across tumor types. Methods: Utilizing the National Cancer Institute’s Mitelman Database (MD) of Chromosome Aberrations and Gene Fusions patients (pts) were identified with EN and categorized based on tumor type, subtype and incidence. Cancer pts who received tumor profiling with Caris were also surveyed for EN. Results: 47 pts with EN across 12 cancer types were extracted from the MD and had median age of 0.17 years (7 unreported); 38% male; 51% acquired malignancies, 49% congenital; 62% cases were pediatric, 23% adult and 15% unknown. 0/204 pts with Caris tumor profiling were found to have an EN. Cancers with the highest number of EN were: 15 (31.9% EN data set) congenital mesoblastic nephromas (CMN), 10 (21.3%) CF, 7 (14.9%) breast carcinoma (BC; 6 secretory ductal carcinoma (SD) and 1 invasive adenocarcinoma (IA)) and 3 (6.4%) colorectal carcinoma (CRC). EN were found in 8 other malignancies (Table 1). Cancer types with the highest incidence of EN+ cases in the MD were gastrointestinal stromal tumor (GIST; 100%), CMN (75%) and CF (23.3%). Conclusions: These results further our understanding of the distribution of ETV6-NTRK3 translocations in multiple tumor types across the age spectrum and suggest that pts with CMN, CF, BC and CRC requiring high order therapy should be considered for NTRK3-based treatment. [Table: see text]
Comparison of response to chemotherapy in non-Hodgkin lymphoma patients depending on their myeloid derived suppressor cell status: Insight on new immune target.
Authors: Mukherjee Kalyan Kusum , Ghosh Sarbari , Halder Subhom , Sarkar Madhurima , Dasgupta Shayani , Chakraborty Mohona , Bose Anamika , Baral Rathindranath
Published: September 2018

e23155

Background: Non Hodgkin’s Lymphoma shows different disease status after completion of standard 1st line treatment. Low grade follicular lymphoma remains stable after standard treatment for a long time and some times it autoregress . In High grade Lymphoma (Commonly diffuse B-cell lymphoma) relapsed 30-40% after 1stline treatment. Immunological target for this differentiation in tumor biology is sought for. Methods: Twenty Five CD20+NonHodgkin's Lymphoma(NHL) patients and one T cell lymphoma were treated with standard chemotherapy (6-8 cycles of RCHOP/CHOP). Within these 25 patients, 10 patients were disease free or exhibited stable disease for two years following the completion of treatment. Rest of the patients showed early relapse (within 6 months) or late relapse (within 18 to 24 months). Eight among 10 disease free patients was suffering from NHL of low grade follicular type. In order to correlate the immune cell status with lymphoma relapse, we have studied a panel of T cells, memory T cells, suppressor cells, after completion of chemotherapy and subsequently in every 6 month interval till relapse. Among several parameters studied a direct correlation between myeloid derived suppressor cells (MDSCs) profile and relapse was noted. Results: CD33+CD11b+MDSCs were remarkably high in high grade NHL patients in comparison to disease free low grade follicular lymphoma patients (p < 0.01). Moreover, all of these NHL patients possessed significantly greater proportion of MDSCs than normal healthy individuals (p < 0.001). Interestingly, single responding T cell lymphoma patient showed completely normal level of MDSCs. Studies are also being conducted to correlate with the status of memory T cells with disease relapse. Conclusions: Observed correlation between greater proportions of MDSCs with the relapse of diseases suggest MDSCs might be the potential target to enhance the relapse free survival of NHL patients, thus, demands further extensive investigation.
CLIA validation workflow of a novel tumor cell isolation platform.
Authors: Sero Valeria , Khurana Aditi , Ting Marc , Webb Judy , Verma Suman , Gerber Amanda , Bolognesi Chiara , Sims Cynthe , Gunn Shelly , Cotter Philip , Moore Mathew , Medoro Gianni , Manaresi Nicolò , Bischoff Farideh Z.
Published: September 2018

e23161

Background: DEPArray™ technology is based on Dielectrophoresis (DEP). High quality image-based cell selection enables users to identify, isolate and recover intact specific individual rare cells of interest from complex, heterogeneous tissues such as live or fixed cell suspensions. Here we demonstrate a validation workflow for application of molecular sequencing technologies and FISH downstream of rare cell recovery. Methods: FFPE scrolls (n = 93) were dissociated and stained with markers to distinguish putative stromal and tumor populations. DNA integrity was tested using a qPCR technique to predict the amount of DNA required for a successful downstream sequencing metrics. DEPArray™ recovered stromal and tumor cells underwent NGS analysis on an Illumina platform based DEPArray™ OncoSeek Panel. Separately, recovered stromal and tumor cells were also utilized for targeted HER2 FISH. CellSearch® cartridges (19 Breast and 5 Bladder Cancers) were processed through the DEPArray to isolate WBCs (CK-/CD45+/DAPI+; n = 30), CTCs (CK+/CD45-/DAPI+; n = 33) and atypical (CK-/CD45-/DAPI+; n = 47) cells. Recovered cells underwent whole-genome amplification with Ampli1™ WGA and quality controlled by Ampli1 QC. Ampli1 LowPass kit was then used to prepare NGS libraries for absolute CNA profiling by low-pass WGS. Results: Reproducibility and reliability was reported as 100% for instrument performance. For FFPE, the OncoSeek panel simultaneously detected SNPs, indels and CNAs of 63actionable and oncology relevant genes. For FISH, 95% concordance with conventional HER2 results was observed. For CTCs, Ampli1 LowPass detected copy number gains/losses confirming tumor origin of the CK+ cells. A portion of non-conventional cells also demonstrated copy number alterations consistent with tumor origin. Conclusions: The isolation of pure single or pooled tumor cells with the DEPArray™ technology can be used as a method to improve downstream MDx analysis using different techniques, thus to inform treatment decisions and provide valuable prognostic information.
Identification of distinctive patterns in cell signaling pathways in glioblastoma multiforme subtypes using gene expression TCGA data sets.
Authors: Volyanskyy Konstantin , Zhong Minghao , Keswarpu Payal , Fallon John T , Fanucchi Michael Paul , Dimitrova Nevenka
Published: September 2018

e23162

Background: Cancer is characterized by a variety of heterogeneous genomic and transcriptomic patterns involving highly complex signaling biological pathways. The problem of identification of the factors driving tumor progression becomes even more challenging due to intricate interaction mechanisms between these pathways. Using novel approaches in machine learning, we demonstrate the ability to quantitatively describe characteristic signaling patterns in cancer based on transcriptomic data Methods: We used RNASeq data from 20531 genes in 174 samples of GBM from The Cancer Genome Atlas including 5 major histological subtypes – Classical, G-CIMP, Mesenchymal, Neural, and Proneural, anddeveloped predictive computational framework for molecular subtype differentiation from normal tissue relying on variance based gene selection and random forest algorithm. Results: We obtained a few key findings – (1) genes from cell signaling pathways alone differentiate each subtype from normal tissue with 100% accuracy; (2) predictive genes are specific to each subtype; (3) inferred pathway interactions are also specific to each subtype; (4) typically most of the predictive genes involved in signaling are down-regulated in tumor compared to normal tissue (MAPT, PRKCG, PDE2A, RYR2, ATP1B1, GRN1, GNAO1), however, in each subtype we observed a smaller subset of predictive genes which are highly up-regulated in tumor (ID3, FN1, JAG1, F2R, COL4A1, EDAR, CDK2, CDK4, MFNG, BIRC5, CCNB2). We detected and quantitatively evaluated characteristic signaling pathway involvement across the GBM subtypes for MAPK, RAP1, RAS, Notch, PI3K-Akt, mTOR, FoxO, Jak-STAT, Wnt, cAMP, and Calcium Signaling, providing a unique approximation for each subtype signaling profile. Conclusions: In this study, we identified gene expression profiles and associated signaling pathways for distinguishing GBM Multiforme subtypes from normal tissue. We observed and described a dense complex picture of interacting signaling pathways. The detected interactions may provide clinical insights and could be used to identify potential therapeutic targets, however, more research is needed to confirm this.
Precision medicine for recalcitrant cancers with the patient-derived orthotopic xenograft (PDOX) mouse models for identification of effective therapy.
Authors: Murakami Takashi , Kawaguchi Kei , Kiyuna Tasuku , Igarashi Kentaro , Miyake Kentaro , Hiroshima Yukihiko , Nelson Scott D. , Dry Sarah M , Li Yunfeng , Singh Arun S. , Eilber Frederick C. , Hoffman Robert M.
Published: September 2018

e23164

Background: Our laboratory has developed the patient-derived orthotopic xenograft (PDOX) mouse model of all major cancer types. Methods: Sarcomas from patients were transplanted into the right bicep femoris muscle of nude mice or at a site corresponding to their metastasis. Melanomas were transplanted in nude mice at the orthotopic site corresponding to their metastasis in the patient. An HER-2-expressing cervical cancer was transplanted to the uterine cervix of nude mice. Results: A Ewing’s sarcoma with a FUS-ERG fusion and a CDKN2A-deletion was resistant to doxorubicin as was the patient and sensitive to palbociclib and linsitinib. A spindle-cell sarcoma PDOX was resistant to doxorubicin, but sensitive to pazopanib. A pleomorphic rhabdomyosarcoma PDOX was found resistant to cyclophosphamide, but sensitive to the combination of temozolomide and irinotecan. A melanoma with a BRAF-V600E mutation PDOX was resistant to vemurafenib even thought it targets the BRAF-V600E mutation and was sensitive to trametinib, but resistant to cobimetinib, although both are MEK inhibitors. The primary tumor in a cervical cancer PDOX was resistant to endostatin, but the metastasis in this model was sensitive. Conclusions: The PDOX models have distinguished differential sensitivity of primary and metastatic tumors to the same drug. PDOX models can also distinguish effective drugs when they are sensitive to first-line therapy. Most importantly, the PDOX models can find potential effective therapeutics for recalcitrant disease. Drug evaluation with the PDOX models demonstrate that mutational analysis of the patient tumor may not be sufficient to direct therapy by itself and together with PDOX testing, can result potentially in precision treatment.
Analyses on expression of trimethylation of histone H3 at lysine 27 and DNA methylation at CCGG sites in smoking and nonsmoking non-small cell lung cancer patients and their clinical significance.
Authors: Chen Xiaohui , Deng Yujie , Zhu Kunshou , Nagayasu Takeshi , Hu Dan , Xu Chunwei , Lin Gen , Huang Cheng , Chen Gang , Zheng Xiongwei , Cai Yibin , Lu Jian-ping , Weng Guibin
Published: September 2018

e23179

Background: Smoking usually leads to epigenetic alterations like DNA methylation and histone modifications. We analyzed the effect that smoking had on DNA methylation level at CCGG sites and expression of trimethylation of histone H3 lysine 27 (H3K27me3), enhancer of zeste homolog 2 (EZH2) and their interactions in non-small cell lung cancer (NSCLC) patients. Methods: 42 NSCLC patients, 22 adenocarcinomas and 20 squamous cell carcinomas were enrolled. Expression of H3K27me3, EZH2 and PCNA were immunohistochemically detected. Apoptotic index in NSCLC with different smoking status was evaluated via TUNEL method. DNA methylation at CCGG sites was evaluated by HELMET method. Their correlation with clinicopathological data were calculated in relation to different smoking status. Results: Compared to the nonsmokers, smoker NSCLC patients were found more often of lower level of DNA methylation at CCGG sites, lower H3K27me3 expression and higher EZH2 expression, and lower apoptotic cell index ( P< 0.05). DNA methylation level at CCGG sites negatively correlated to the Binkman Index( P= 0.017). Further, trend of H3K27me3 and EZH2 expression was in the same direction in most smokers, while in nonsmokers, their expression trend was mostly in the opposite direction ( P= 0.015). Trend of PCNA and EZH2 expression was in the opposite direction in most smokers, while in nonsmokers, their expression trend was mostly in the same direction ( P= 0.048). In addition, trend of CCGG methylation ratio and immunohistochemical expression of H3K27me3 was in the same direction in most smokers, while in nonsmokers, the trend was mostly in the opposite direction ( P= 0.049). Conclusions: NSCLC patients of different smoking status exhibit completely different epigenetic characteristics, and DNA methylation at CCGG sites would probably determine the expression trend of H3K27me3, EZH2 and PCNA.
Sex hormones as regulators of levels of growth factors in tissues of multiple primary tumors of female reproductive organs.
Authors: Kit Oleg Ivanovich , Frantsiyants Elena Mikhaylovna , Moiseenko Tatiana , Vereskunova Marina I. , Adamyan Meri Viktorovich , Saforian Natalya S. , Gurnak Viktor Viktorovich , Ilchenko Maria G.
Published: September 2018

e23180

Background: Sex hormones are known to be able to influence the synthesis of growth factors in breast and uterine tumors; however, their relationship in multiple primary tumors is poorly studied. The purpose of the study was to analyze correlations between sex steroids and growth factors in tissues of solitary and synchronous breast cancer and uterine myoma. Methods: Levels of estrogens (E1, E2, E3), 2OHE1 and 16α-OHE1, Р4, VEGА and EGF were measured by ELISA in tissues of breast cancer (BC, n = 37), uterine myoma (n = 27) and in synchronous BC and uterine myoma (n = 36). Results: Strong positive correlations were observed in synchronous BC and uterine myoma: E3 with VEGF (r1= 83, r2= 79; p1,2< 0.05) and EGF (r1= 78, r2= 82; p1,2< 0.05), Е1 with VEGF ( r1= 81, r2= 77; p1,2< 0.05) and EGF (r1= 76, r2= 80; p1,2< 0.05), 16-OHE1 with VEGF and EGF (r1= 80, r2= 77; p1,2< 0.05 and r1= 79 r2= 80; p1,2< 0.05). We found strong negative correlations of Е3 and VEGF (r1= 80, r2= 77; p1,2< 0.05) and EGF (r1= 79 r2= 80; p1,2< 0.05), Р4 and VEGF (r1= - 81, r2= -79; p1,2< 0.05) and EGF (r1= - 77, r2= -80; p1,2< 0.05). Solitary breast tumors showed moderate positive correlations of E2 and E1 with VEGF (r1= 56, r2= 63; p1,2< 0.05) and EGF (r1= 61, r2= 65; p1,2< 0.05) and a moderate negative correlation between VEGF and EGF and Р4 (r1= - 61, r2= -59; p1,2< 0,05). A strong negative correlation of EGF with Е1 and 16-OHE1 (r1= -79, r2= -76; р1,2< 0.05) was registered in uterine myoma. Conclusions: Tissues of synchronous primary breast tumors and uterine myoma had similar correlations between levels of growth factors and sex hormones. Solitary breast tumors and uterine myoma showed correlations typical of the tumor type. Thus, expression of growth factors in multiple primary tumors, but not in solitary ones, is subject to a clearly determined regulatory role of sex steroids and their metabolites.
JAK2 variations and functions in lung adenocarcinoma.
Authors: Yanjun Xu , Yun Fan
Published: September 2018

e23181

Background: Lung cancer ranks as the first most common cancer and the first leading cause of cancer-related death in China and worldwide. Due to the difficulty in early diagnosis and the onset of cancer metastasis, the 5-year survival rate of lung cancer remains extremely low. JAK2 has emerged as pivotal participant in biological processes, often dysregulated in a range of cancers, including lung cancer. Recently we found that JAK2 might play an important role in lung cancer pathogenesis as an oncogene. While our understanding of JAK2 in the onset and progression of lung cancer is still in its infancy, there is no doubt that understanding the activities of JAK2 will certainly secure strong biomarkers and improve treatment options for lung cancer patients. Methods: The expression level of JAK2 mRNA was assayed using RT-PCR. JAK2 mutations and amplification were detected using next-generation sequencing (NGS). MTT assay, Transwell migration and invasion assay were conducted to study the proliferation, migration and invasion abilities of lung adenocarcinoma cells independently. The shRNA and overexpression plasmids of JAK2 were conducted. Results: JAK2 is up-regulated in lung adenocarcinoma tissues when compared with their adjacent non-tumor tissues, and was associated with lymph node metastasis ( p< 0.05). JAK2 V617F and N30S mutations and JAK2 amplification were detected by NGS in lung adenocarcinoma patient samples. Downregulation of JAK2 inhibits the proliferation, migration and invasion abilities of lung cancer cells. Moreover, overexpression of JAK2 induced the proliferation, migration and invasion abilities of lung cancer cells. Conclusions: These findings demonstrate that JAK2, whose expression is up-regulated in lung adenocarcinoma, whose mutation and amplification were detected in lung adenocarcinoma, may participate in lung cancer progression by regulating cancer cells proliferation, migration and invasion.
Validation of the AJCC 8th edition prognostic stage in breast cancer.
Authors: Weiss Anna , Chavez-Mac Gregor Mariana , Lichtensztajn Daphne , Yi Min , Clarke Christina A , Giordano Sharon Hermes , Hunt Kelly , Mittendorf Elizabeth Ann
Published: September 2018

e23186

Background: The 8th edition of the AJCC staging manual introduced a prognostic stage incorporating biologic factors. The goal this study was to validate the prognostic stage in a single institution cohort and large population database. Methods: Patients were identified in a prospective institutional database and the California Cancer Registry (CCR). Factors included were T, N, m stage, grade, estrogen receptor, progesterone receptor and HER2 status. Survival was calculated by Kaplan Meier method. Harrell’s concordance index (C index) was used to quantify models’ predictive performance, and Akaike Information Criterion (AIC) to compare model fits. Results: A total of 3,327 Stage I-IIIC breast cancer patients treated 2007-2013 at a single institution with complete clinicopathologic data were identified. Compared to the anatomic stage, the prognostic stage upstaged 29.5% of patients and downstaged 28.1%. Median follow up was 5 yrs. The prognostic staging system’s C index (0.84) and AIC (816.8) reflect a more accurate model predictive of disease specific survival (DSS) (Table) than the anatomic staging (C index 0.74, P < .0001; AIC 1039.8). A total of 49,767 Stage I-IIIC patients diagnosed 2005-2009 with complete data were identified in the CCR. 31.7% of patients were upstaged and 21.1% downstaged. Median follow up was 7.15 years. Prognostic staging performed better (C index 0.81, AIC 65924.1) than anatomic staging (C index 0.77, P < .0001; AIC 66847.7). Conclusions: The newly released 8th AJCC prognostic staging system, validated in both a single institution and a large population database, provides more accurate prognostic information than the anatomic staging system. [Table: see text]
A genomic signature based in IGF1R-network in breast cancer.
Authors: Gomez Andrea C. , Araujo Jhajaira M. , Flores Claudio J. , Velazco Rodolfo , Gomez Henry Leonidas , Pinto Joseph A.
Published: September 2018

e23187

Background: The insulin-like growth factor 1 receptor (IGF1R) is a member of the receptor tyrosine kinase family and has been involved in tumor transformation and survival of malignant cells. The evaluations of interactions networks of this protein could be helpful to find new therapeutic targets and prognostic factors. Our aim was to develop a genomic signature for disease-free survival (DFS) based on IGF1R signaling. Methods: We identified in STRING v.10.0 a set of proteins participating in the network of IGF1R. Analysis was done with a high confidence (0.7). We identified GEO datasets of microarray experiments containing gene expression profiling of breast tumors. The end point was DFS. We use one discovery set (GSE25066, N = 508) and two validations sets (GSE42568, N = 121; GSE1456, N = 318). Univariate Coxph identified a first set of genes that were screened in a multivariate analysis and then evaluated in Km-plotter (http://kmplot.com). Genes significantly associated with the DFS were analyzed in a multivariate Coxph to estimate regression coefficients to develop a linear score. Results: Analysis in STRING identified 100 proteins whose coding genes were evaluated in a first univariate analysis identifying 30 genes. The multivariate analysis identified 5 genes associated with the DFS (VCL, IARS, IRS1, IGF1R and MAKP3). The bioinformatics platform Km-plotter selected IRS1, IARS, IGF1R and MAKP3 as predictors for DFS. The multivariate analysis with the stepwise method selected only three genes, IARS HR = 147.82 (P < 0.001, 95% CI: 11.60-1883.35); IGF1R HR = 0.11 (P < 0.001, 95% CI: 0.04-0.31) and MAKP3 HR = 0.36 (P = 0.012, 95% CI: 0.16-0.80) as significantly associated with the DFS. With the regression coefficients, we developed the follow signature: (-1.026 x MAPK3) + (-2.250 x IGF1R) + (4.996 x IARS). The median score in the discover set (0.544) was able to identified patients at different risk of recurrence (P < 0.001 in the Logrank test), using the median using as cutoff the signature was able to discriminate two groups of different risk of recurrence in GSE42568 and GSE1456 validation sets (P > 0.02 and P > 0.03, respectively). Conclusions: We were able to develop a genomic predictor based in the network of IGF1R. In ours study IGF1R was associated to good prognosis.
Association of activin type II receptor mutation with microsatellite instability in gastric cancer.
Authors: Yuza Kizuki , Nagahashi Masayuki , Ichikawa Hiroshi , Nakajima Masato , Hanyu Takaaki , Ishikawa Takashi , Shimada Yoshifumi , Sakata Jun , Kameyama Hitoshi , Kobayashi Takashi , Nakagawa Satoru , Sato Nobuaki , Honma Keiichi , Kawasaki Takashi , Okuda Shujiro , Takabe Kazuaki , Wakai Toshifumi
Published: September 2018

e23191

Background: Microsatellite instability-high status (MSI-H) and alterations in the DNA mismatch repair pathway associate with the efficacy of 5-FU and immune checkpoint inhibitors in patients with gastrointestinal cancers. The activin type II receptor (ACVR2) that binds to the transforming growth factor beta superfamily of ligands is frequently mutated in MSI-H colorectal cancer. However, the incidence of ACVR2 mutations in gastric cancer patients remains unclear. The aim of this study is to reveal the incidence and to examine the association between the MSI-H and ACVR2A mutations in gastric cancer patients. Methods: 124 archived FFPE gastric cancer tissues (stage I-IV), who were operated at Niigata University Medical and Dental Hospital or Niigata Cancer Center Hospital, were analyzed for ACVR2A mutation and MSI status with the NGS-based comprehensive genomic test platform. Clinicopathological characteristics of the patients were also examined. Results: All 124 gastric cancer patients were successfully analyzed. 13 out of 124 patients (10.4%) showed MSI-H status. Interestingly, 10 of 13 MSI-H patients (76.9%) showed ACVR2A mutation, where none (0%) was found among patients with microsatellite stable status (P < 0.001), indicating the strong association between ACVR2A mutation and MSI status in gastric cancer patients. In the ACVR2A mutated group, there was a female predominance (P < 0.05), and cancers of the lower part of the stomach were more common (P < 0.05), compared with the wild type group. Only one of 10 patients with ACVR2A mutation died, and the patients with ACVR2A mutation show a 5-year overall survival rate of 90%. No statistically significant difference in survival was achieved between patients with ACVR2A mutation and wild type; this is probably due to the small number of patients. Conclusions: 10 of 13 MSI-H patients showed ACVR2A mutation. Our results indicate a strong association between ACVR2A mutation and MSI-H in gastric cancer patients.
Ginsenoside Rg3 to regulate the EMT induced by TGF-in gastric cancer.
Authors: Zhao Lianjun , Lv Qing , Xv Qiuping , Shao Jie , Su Shu , Meng Fanyan , Wei Jia , Liu Baorui , Qian Xiaoping , Yu Lixia , Zou Zhengyun
Published: September 2018

e23200

Background: To discuss the regulate effect of Ginsenoside Rg3 during the TGF-β1 induced EMT in human gastric cancer SNU-601 cell line. Methods: In the first part of our experiment, ninety-nine patients who were diagnosed as gastric cancer were admitted to this study eventually. The Luminex® xMAP technology was used to measure the plasma levels of TGF-β1. In the second part, human gastric cancer SNU-601 cell line were cultured normally. Inverted microscope was used to observe morphological changes. The cell relative growth rate of proliferation was analyzed by MTT assay. Wound-healing assay and transwell invasion assay were carried out to predict the capacity of Ginsenoside Rg3 in inhibiting migration and invasion. Western Blot was performed to analyze the influence of Ginsenoside Rg3 and TGF-β1 on the protein expression of E-cadherin and Vimentin. Results: The plasma cytokine levels of TGF-β1 were associated with vessel invasion, peritoneal involvement, TNM stage, T stage, N stage in gastric cancer patients. In Vitro Study, TGF-β1 stimulated epithelial-mesenchymal transition in SNU-601 cell lines. Ginsenoside Rg3 inhibited the TGF-β1 induced morphological changes, cell proliferation increase, cell migration and cell invasion. Ginsenoside Rg3 markedly increased expression of the epithelial marker E-cadherin, and repressed the upregulation and expression of the mesenchymal marker Vimentin during the TGF-β1 induced EMT in SNU-601 cell line. Conclusions: The plasma cytokine levels of TGF-β1 were associated with vessel invasion, peritoneal involvement, tumor stage in gastric cancer patients. TGF-β1 stimulated biological malignant behavior through EMT in gastric cancer. Ginsenoside Rg3 could inhibit the EMT induced by TGF-β1 in SNU-601 cell lines.
Performance of a high-intensity 508-gene circulating-tumor DNA (ctDNA) assay in patients with metastatic breast, lung, and prostate cancer.
Authors: Razavi Pedram , Li Bob T. , Abida Wassim , Aravanis Alex , Jung Byoungsok , Shen Ronglai , Hou Chenlu , De Bruijn Ino , Gnerre Sante , Lim Raymond S. , Reales Dalicia , Maddala Tara , Berger Michael F. , Riely Gregory J. , Scher Howard I. , Novotny William F. , Solit David B. , Lee Mark , Reis-Filho Jorge S. , Baselga Jose
Published: September 2018

LBA11516

The full, final text of this abstract will be available at abstracts.asco.org at 7:30 AM (EDT) on Saturday, June 3, 2017, and in the Annual Meeting Proceedings online supplement to the June 20, 2017, issue of the Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Saturday edition of ASCO Daily News.
Durable remissions with BCMA-specific chimeric antigen receptor (CAR)-modified T cells in patients with refractory/relapsed multiple myeloma.
Authors: Fan Frank (Xiaohu) , Zhao Wanhong , Liu Jie , He Aili , Chen Yinxia , Cao Xingmei , Yang Nan , Wang Baiyan , Zhang Pengyu , Zhang Yilin , Wang Fangxia , Lei Bo , Gu Liufang , Wang Xugeng , Zhuang Qiuchuan , Zhang Wanggang
Published: September 2018

LBA3001

The full, final text of this abstract will be available at abstracts.asco.org at 7:30 AM (EDT) on Monday, June 5, 2017, and in the Annual Meeting Proceedings online supplement to the June 20, 2017, issue of the Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Monday edition of ASCO Daily News.
Phase 1/2 study of the selective TRK inhibitor larotrectinib in pediatric patients with cancer.
Authors: Federman Noah , Albert Catherine Michelle , Turpin Brian , Mascarenhas Leo , Nagasubramanian Ramamoorthy , Geoerger Birgit , Casanova Michela , Gallego Melcon Maria Soledad , Chisholm Julia , Van Tilburg Cornelis M. , Shukla Neerav Narendra , Spunt Sheri L. , Cox Michael Craig , Hawkins Douglas S. , Pappo Alberto S. , Bielack Stefan S. , Doz Francois P. , Bisogno Gianni , Laetsch Theodore Willis , DuBois Steven G. ,   Larotrectinib SCOUT Investigators
Published: September 2018

TPS10577

Background: Neurotrophin ligands and their receptors TRKA, TRKB, and TRKC (encoded by NTRK1, NTRK2, and NTRK3) are important for growth regulation, differentiation and survival of neurons. Translocations involving the NTRK1/2/3 kinase domain, mutations involving the TRK ligand-binding site, and amplifications of NTRK, have been described in diverse tumor types and may contribute to tumorigenesis. A broad range of pediatric malignancies have been found to harbor NTRK fusions, including infantile fibrosarcoma (IFS), spindle-cell sarcoma, congenital mesoblastic nephroma, pediatric papillary thyroid cancer, pediatric gliomas and Ph-like acute lymphoblastic leukemia. Larotrectinib is the first small-molecule selective inhibitor of TRKA, -B, and -C in clinical development and preliminary data from the adult phase 1 trial demonstrate prolonged responses in patients with TRK fusions and a favorable safety profile. Methods: We have initiated an open-label, multi-center, international Phase 1/2 study with larotrectinib in pediatric patients with solid tumors and primary CNS tumors (NCT02637687). Patients with advanced cancer between the ages of 1 year and 21 years are eligible, as well as patients as young as 1-month of age with a documented NTRK fusion. Patients with IFS who have not had definitive surgery are also eligible. Larotrectinib is administered orally twice daily on a continuous 28-day schedule. Dosing is based on body surface area. Larotrectinib is available in an oral liquid formulation and capsules. Following identification of the maximum tolerated dose of larotrectinib in the phase 1 portion, the phase 2 portion will commence. The phase 2 portion will enroll patients with NTRK-translocated tumors and measurable disease into three cohorts: 1) infantile fibrosarcoma; 2) extracranial solid tumors; and 3) primary CNS tumors. The primary endpoint for the phase 2 portion is objective response rate, with duration of response and progression free survival as secondary efficacy endpoints. Each phase 2 cohort will enroll in a single stage of up to 10 patients per cohort. Molecular abnormalities will be characterized through the analysis of archival tissue. Enrollment began in December 2015 and is ongoing. Clinical trial information: NCT02637687.
Comparative genomic analysis for pediatric cancer patients evaluated in a California Initiative to Advance Precision Medicine Demonstration Project.
Authors: Morozova Olena , Salama Sofie R. , Bjork Isabel , Goldstein Theodore C. , Mueller Sabine , Sender Leonard S. , Sweet-Cordero Alejandro , Haussler David ,   California Kids Cancer Comparison Team
Published: September 2018

TPS10578

Background: California Kids Cancer Comparison (CKCC), a demonstration project for the California Initiative to Advance Precision Medicine, evaluates the utility of incorporating gene expression information into the genomic analysis of difficult-to-treat pediatric cancers. CKCC is a partnership between UC Santa Cruz and clinical genomic trials conducted by Children’s Hospital of Orange County, UC San Francisco (Pacific Pediatric Neuro Oncology Consortium), and Stanford University. Methods: CKCC compares each prospective tumor’s RNA sequencing profile to over 11,000 uniformly analyzed tumor profiles from pediatric and adult cancer patients. These comparisons are used to identify genes and pathways that are significantly over expressed in each patient’s tumor. The pathways are reviewed by data analysis for the potential for clinical impact and presented to the treating oncologist in a molecular tumor board setting.
A randomized phase II trial of fulvestrant with or without ribociclib after progression on aromatase inhibition plus cyclin-dependent kinase 4/6 inhibition in patients with unresectable or metastatic hormone receptor positive, HER2 negative breast cancer.
Authors: Kalinsky Kevin , Mundi Prabhjot Singh , Chiuzan Codruta , Accordino Melissa Kate , Trivedi Meghna S. , Sparano Joseph A. , Young Oh Sun , Tiersten Amy , O'Regan Ruth , Esteva Francisco J. , Jain Sarika , Mayer Ingrid A. , Forero Andres , Baer Lea N. , Crew Katherine D. , Hershman Dawn L. , Vaklavas Christos
Published: September 2018

TPS1112

Background: CDK4/6i, including palbociclib and ribociclib (R), have demonstrated remarkable benefit in progression free survival (PFS) in patients (pts) with HR+, HER2- MBC with anti-estrogen therapy. Switching between anti-estrogen therapies at disease progression is standard of care in the treatment of HR+ MBC. We evaluate the strategy of switching anti-estrogen therapy to fulvestrant (F) and maintaining CDK4/6 inhibition with R in pts with HR+, HER2- MBC who have progressed on an AI + CDK4/6i. Methods: Trial Design Phase II, multi-center, randomized, double-blind, placebo-controlled trial to evaluate F +/- R in pts with HR+, HER2- MBC who have previously progressed on any AI + CDK4/6i: Screened at 2 different scenarios: Scenario 1: Before receiving any CDK4/6i Scenario 2: Time of progression of disease (POD) while being treated with an AI + CDK4/6i Intervention At randomization, pts assigned 1:1 to either a) F + R or b) F + placebo, with treatment given in 4-week cycles. Major Eligibility Criteria 1, Metastatic BC, 2. HR+ HER2-, 3. Measurable or unmeasurable disease Specific Aims Primary: PFS. Secondary: Objective response rate, clinical benefit rate, overall survival, and duration of response. Biomarker assessment: amplification of cyclin D1 and cyclin E, phosphoRb and TK1 expression, Rb1 and p16 loss, and ctDNA for ESR1 and PIK3CA mutations. Statistical Methods Assuming a median PFS of 3.8 months with F alone, we predict that F + R will lead to a median PFS of at least 6.5 months. A one-sided log-rank test with a sample size of N = 120 and alpha = 0.025, achieves 80% power to detect a difference in PFS of 2.7. With a 10% dropout, n = 132. Clinical trial registry number NCT02632045.