bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 588: Neutrophil to lymphocyte ratio as strongest predictor of early death in metastatic triple negative breast cancer
Authors: Ku Gabriel A. De la Cruz , Morante Zaida , Pinto Joseph , Araujo Jhajaira , Enriquez Daniel , Eyzaguirre Eduardo , Saavedra Antonella , Lujan Maria , Luque Renato , Rázuri Cesar , Fuentes Hugo , Neciosup Silvia , Moreno Henry Gómez
Published: August 2018

Background: Metastatic triple-negative breast cancer (mTNBC) is a heterogeneous disease with poor prognosis where recent studies suggest that the immune system plays a critical role the mechanisms of disease progression and metastasis. In the other hand, a high Neutrophil to Lymphocyte Ratio (NLR) is associated with poor survival in patients diagnosed with several types of cancer. Our aim was to evaluate the prognostic value of NLR in mTNBC.

Methods: We retrospectively reviewed 130 patients diagnosed with mTNBC between 2000-2014 at “Instituto Nacional de Enfermedades Neoplasicas”. NLR group was divided in <2.5 and ≥2.5. Sites of metastasis were identified by TAC or magnetic resonance imaging. Survival rates were estimated by the Kaplan-Meier method and differences were calculated by Log-rank test. The Cox Regression was used for prognostic factors.

Results: The median age was 50 years (range: 23-92 years). From our study population, 41.2% were premenopausal, and just 10 patients (9.5%) had a breast-ovary cancer familiar background, 62.5% were diagnosed at clinical stage T3-T4 and 86.1% had lymph node involvement. From 130 patients, 41 and 64 had NLR <2.5 and ≥2.5, respectively, while there was no association between characteristics and NLR, only patients with malnutrition had NLR ≥2.5 (9.1% vs 40.8%, p=0.002) and solid organ involvement (62.4% vs 81.3%, p=0.04). Furthermore, lung and bone were the most frequent sites of metastasis. Those patients with lower ratio (<2.5) had better response to chemotherapy (p <0.001). With a median follow-up of 6.25 years, at 1-year OS was lower among patients with NLR ≥2.5 (59% vs 20%, HR: 2.77, 95%CI:1.77-4.33, p<0.001). In addition, at univariate analysis malnutrition also showed an impact on OS (HR: 1.79, 95%CI: 1.20-2.67, p=0.005). At multivariate analysis, between age, T stage, N stage, NLR and solid organ involvement, just NLR was the best and only predictor of worse OS (HR: 3.12, 95%CI:1.94-5.03, p<0.001). 

Conclusion: In mTNBC, an NLR ≥2.5 was a strong and accessible biomarker of poor OS. Lower values of NLR could help us to predict chemotherapy response.

Citation Format: Gabriel A. De la Cruz Ku, Zaida Morante, Joseph Pinto, Jhajaira Araujo, Daniel Enriquez, Eduardo Eyzaguirre, Antonella Saavedra, Maria Lujan, Renato Luque, Cesar Rázuri, Hugo Fuentes, Silvia Neciosup, Henry Gómez Moreno. Neutrophil to lymphocyte ratio as strongest predictor of early death in metastatic triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 588.
Abstract 5880: Small-molecule modulators of PDE3/SLFN12 to kill cancer cells
Authors: Lewis Timothy A. , Waal Luc de , Wu Xiaoyun , Ellerman Manuel , Kopitz Charlotte , Wengner Antje , Eis Knut , Lange Martin , Tersteegen Adrian , Lienau Philip , Greulich Heidi , Meyerson Matthew
Published: August 2018

We have previously reported1 our results with a small molecule called DNMDP which kills certain cancer cells by modulating the interaction of PDE3A and Schlafen 12 (SLFN12), a more recently discovered protein of unknown function. While DNMDP selectively inhibits PDE3, most PDE3 inhibitors have no cell killing effects and in fact rescue cancer cells from DNMDP-induced death. DNMDP is not suitable for advanced studies due to structural liabilities. Optimization of DNMDP to both increase activity and improve pharmacokinetic properties resulted in enantiomerically pure, low molecular weight, metabolically stable compounds which are active at low doses in animal models of cervical cancer and melanoma. While these compounds are selective PDE3A inhibitors and their biochemical activity mirrors their cellular activity, the activity of the compounds results not from PDE3 inhibition and increased cAMP levels, but from increased compound-induced binding of PDE3A to SLFN12, which most PDE3 inhibitors do not effect. Our results suggest that small molecule modulators of PDE3/SLFN12 binding may provide a novel treatment for the treatment of certain cancers.

1Nat. Chem. Bio. 2016, 12, 102-108.

Citation Format: Timothy A. Lewis, Luc de Waal, Xiaoyun Wu, Manuel Ellerman, Charlotte Kopitz, Antje Wengner, Knut Eis, Martin Lange, Adrian Tersteegen, Philip Lienau, Heidi Greulich, Matthew Meyerson. Small-molecule modulators of PDE3/SLFN12 to kill cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5880.
Abstract 5896: Downregulation of calcium-sensing receptor enhances MET-mediated prostate cancer chemosensitivity
Authors: Huang Luying , Wang Fuhao , Tong Haibo , Mizokami Atsushi , Keller Evan T. , Lu Yi , Zhang Jian
Published: August 2018

Chemoresistance, in various types of cancer including prostate cancer (PCa), is one of the major challenges in cancer therapy. Others and our lab have reported that the epithelial-to-mesenchymal transition (EMT) plays critical roles in development of chemoresistance and metastasis. The calcium-sensing receptor (CaSR) plays important roles in maintenance of calcium homeostasis and altered CaSR expression, and functional changes are involved in certain diseases, including tumorigenesis and its development. However, the mechanism of CaSR in PCa chemoresistance remains unclear. In current study, the CaSR expression level was detected in parental PC3 and DU145 cells and their chemoresistant PC3-TxR and DU145-TxR cells. We found the CaSR was highly expressed in chemoresistant PC3-TxR and DU145-TxR cells. Then by using siRNA technique, CaSR expression was transiently knocked down in chemoresistant cells. The tumor cell migration, invasion, and colony formation were evaluated by wound healing, transwell, and colony formation assays, respectively. The drug sensitivity was evaluated using MTS assay. We observed that the abilities of tumor cell migration and invasion were significantly diminished in CaSR knockdown PC3-TxR and DU145-TxR cells. The ability of colony formation was also reduced. Interestingly, the sensitivity to paclitaxel was reactivated in CaSR knockdown PC3-TxR and DU145-TxR cells. We finally observed that E-cadherin expression was increased in CaSR knockdown chemoresistant cells, and Snail, Vimentin and β-catenin were decreased; these results are consistent with EMT phenotypic alterations. Taken together, we conclude that downregulation of CaSR enhances MET-mediated prostate cancer chemosensitivity.

Supported by NSFC Key Project 81130046; NSFC projects 81773146,81272415; JCYJ20170412152943794, JCYJ20170412154619484, JCYJ20170307105128101, JCYJ2017030711041760; Guangxi Key Projects 2013GXNSFEA053004.

Citation Format: Luying Huang, Fuhao Wang, Haibo Tong, Atsushi Mizokami, Evan T. Keller, Yi Lu, Jian Zhang. Downregulation of calcium-sensing receptor enhances MET-mediated prostate cancer chemosensitivity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5896.
Abstract 590: Marital status and survival among breast cancer patients
Authors: Lu Yan , Jackson Bradford E. , Ghabach Bassam , Ojha Rohit P.
Published: August 2018

Background: Recent evidence suggests improved survival for married breast cancer patients compared with unmarried patients, but unclear is whether this survival benefit extends to socioeconomically disadvantaged women. We aimed to assess the association between marital status and breast cancer survival among socioeconomically disadvantaged breast cancer patients and breast cancer patients from the general population.

Methods: We used data from the John Peter Smith (JPS) Oncology Registry, which comprises cancer patients in Tarrant County, TX treated at a safety-net institution, and the Texas Cancer Registry (TCR), a statewide population-based registry. Our eligible population included females aged 18 – 64 years diagnosed with primary breast cancer between January 2008 and June 2013 with follow-up through 2016. We categorized marital status into three groups: single, married, or separated/divorced/widowed. We estimated the 36-month restricted mean survival time, which allows survival comparisons within a specified period.

Results: Our study populations comprised 614 JPS and 32,985 TCR patients. The age distribution at diagnosis was similar for both groups (JPS: median=51 years; TCR: median=52 years). JPS patients were more often non-White (JPS=62%; TCR=42%) and uninsured (JPS=56%; TCR=9.6%). A higher frequency of TCR patients were married (TCR=65%; JPS=39%). Survival was shorter for JPS patients (JPS=33.1 months; TCR =34.8 months) after 36-months (Table 1). Married women had longer survival than unmarried women in both populations.

Conclusion: Our results suggest similar magnitudes of improved survival for married women with breast cancer compared with unmarried women in both populations, but socioeconomically disadvantaged patients have consistently lower survival estimates across marital subgroups compared with patients from the general population. Our findings warrant further studies that explore mechanisms of how marital status influences breast cancer survival in different populations.

Table 1. Restricted mean survival time (RMST) after breast cancer diagnosis based on marital statusDisadvantaged Cancer PatientsaCancer Patients from General PopulationbRMSTc (months)Difference in RMST (95% CLd) (months)RMST (months)Difference in RMST (95% CL) (months)Separated/Divorced/Widowed32.2-0.89 (-2.8, 1.0)34.10.01 (-0.23, 0.25)Married33.80.56 (-0.96, 2.1)35.10.54 (0.36, 0.71)Single33.00 (Reference)34.10 (Reference)aCancer patients from JPS Oncology RegistrybCancer patients in the Texas Cancer RegistrycBased on 36-month restricted mean survivaldCL: Confidence limits

Citation Format: Yan Lu, Bradford E. Jackson, Bassam Ghabach, Rohit P. Ojha. Marital status and survival among breast cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 590.
Abstract 5901: The role of POU3F2 in the aggressive behavior of neuroendocrine prostate cancer
Authors: Kobayashi Naoko , Hong Dai , Wang Chun , Yamashiro Joyce , Guan Johnny , Reiter Robert E.
Published: August 2018

Neuroendocrine and Small Cell Prostate Cancer (NEPC) is a lethal disease emerging from Castrate Resistant Prostate Cancer (CRPC) after treatment with next-generation anti-androgen therapies such as abiraterone and enzalutamide. The mechanisms by which treatment resistant prostate epithelial cancer cells trans-differentiate to poorly differentiated small cell cancer are not well understood. We and others found that Nervous System-Specific Octamer-Binding Transcription Factor (POU3F2) expression is induced in NEPC. POU3F2 is a reprogramming transcription factor that controls cellular differentiation and can induce stem-like cells from differentiated glioblastoma. POU3F2 also plays a critical role in melanoma progression and in small cell carcinoma of the lung. NEPC shares similarities with small cell carcinoma from other organs. We hypothesized that POU3F2 may be a key transcription factor that is required for transdifferentiation and the survival of NEPC cells. We further hypothesize that the underlying mechanism of therapy resistance in NEPC is that NEPC consists of stem-like cells that were reprogrammed from differentiated prostate cancer cells under the selective pressure of androgen receptor (AR) pathway-targeted therapy. We elucidated the functional role of POU3F2 in the aggressive cancer behavior of NEPC —including stem-cell properties, tumorigenesis, invasion, and chemo-resistance. We also characterized the mechanisms that regulate POU3F2 expression in therapy-resistant NEPC. The study provides a rational for targeted therapy for treatment of NEPC. It could also be used to design a strategy to halt the emergence of NEPC from CRPC.

Citation Format: Naoko Kobayashi, Dai Hong, Chun Wang, Joyce Yamashiro, Johnny Guan, Robert E. Reiter. The role of POU3F2 in the aggressive behavior of neuroendocrine prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5901.
Abstract 5903: The molecular mechanism for producing BQ323636.1 in Tamoxifen resistance breast cancer cells
Authors: Mok Ka Chun , Tsoi Ho , Khoo Ui-Soon , Man Ellen
Published: August 2018

Breast cancer is one of the prevalent causes of cancer in women. Two thirds of breast cancer patients are ER-positive and can be benefited from tamoxifen treatment. However, 50% of the patients will eventually develop the resistance. Identifying molecular targets associated with tamoxifen resistance would help in designing better therapeutic strategies. Previous study from our group identify a novel factor BQ323636.1 (BQ), a spliced variant of NCOR2, could confer tamoxifen resistance. From our in vivo study, nuclear enrichment of BQ in primary breast tumor was found to be associated with tamoxifen resistance. However, the molecular mechanism governed the nuclear import of BQ and was not clear. From bioinformatics prediction, we identified a functional nuclear localization signal (NLS) on BQ. We therefore performed a protein screening to search for the import receptor which could interact with BQ. KPNA1 was found to interact with BQ. To further validate such an interaction, we fused the NLS to GPF reporter protein (GFP-NLS) and performed a co-immunoprecipitation. The result showed that the GFP-NLS was able to interact with KPNA1. Knockdown of KPNA1 by RNAi could prevent the nuclear import of BQ. This confirmed the functional role of KPNA1 in the nuclear import of BQ. In addition, knockdown of KPNA1 in BQ overexpressing breast cancer cells could reverse the tamoxifen resistance. The results highlighted the functional significance of KPNA1 in the tamoxifen resistance. Subsequently, we investigated the functional role of BQ in the nucleus. We found BQ could deplete the gene repressive function of NCOR2. We found that NCOR2 could interact with HSF4 and should repress the expression of HIF1α. In the presence of BQ, the repressive function of NCOR2 on HIF1α was compromised, leading to up-regulation of HIF1α. To further validate the effect of HIF1α in the tamoxifen resistance, HIF1α inhibitor was employed and found that the treatment of the inhibitor could reduce the resistance in BQ overexpressing breast cancer cells. Therefore, our study uncovered the role of KPNA1 in the development of tamoxifen resistance through which KPNA1 could promote the nuclear import of BQ. Subsequently, BQ could lead to the up-regulation of HIF1α which also play significant role in the development of tamoxifen resistance.

Citation Format: Ka Chun Mok, Ho Tsoi, Ui-Soon Khoo, Ellen Man. The molecular mechanism for producing BQ323636.1 in Tamoxifen resistance breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5903.
Abstract 5908: Resistance mechanisms to TP53-MDM2 inhibition identified by in vivo piggyBac transposon mutagenesis screen
Authors: Chapeau Emilie A.
Published: August 2018

Inhibitors of MDM2-TP53 interaction are predicted to be effective in tumors where the TP53 gene is wild-type by preventing TP53 protein degradation. One such setting is represented by the frequent CDKN2A deletion in human cancer that, through inactivation of p14ARF, activates MDM2 protein, which in turn degrades TP53 tumor suppressor. Here we utilized piggyBac transposon insertional mutagenesis to anticipate resistance mechanisms that occur under intermittent high dose treatment with the MDM2-TP53 inhibitor HDM201. Constitutive piggyBac mutagenesis in Arf-/- mice provided a collection of spontaneous tumors with characterized insertional genetic landscapes. Tumors were allografted in large cohorts of mice to assess HDM201 pharmacological effects. 16 out of 21 allograft models were sensitive to HDM201 but ultimately relapsed under treatment. When tumors with acquired resistance to HDM201 were compared to untreated tumors, 87 genes were found differentially and significantly targeted by piggyBac transposon. Resistant tumors displayed a complex clonality pattern suggesting emergence of several resistant subclones. Among the most frequent alterations conferring resistance, we observed somatic and insertional loss-of-function mutations in Trp53 in 54% of tumors, and transposon-mediated gain-of-function alterations in Bcl-xL, Mdm4, and two TP53-family members, resulting in expression of the TP53 dominant negative truncations ΔNTrp63 and ΔNTrp73. Enhanced BCL-xL and MDM4 proteins expression was confirmed in resistant tumors, as well as in HDM201 resistant patient-derived tumor xenografts. Interestingly, concomitant inhibition of MDM2 and BCL-xL demonstrated significant synergy in p53 wild-type cell lines in vitro.

Moreover, we performed an additional piggyBac screen with daily HDM201 treatment, at lower dose. Surprisingly, we identified Bcl-XL expression as a unique mechanism of resistance in intermittent high dose scheduling. These data suggested that different apoptotic inductions between the two HDM201 dose regimens (intermittent high dose/daily lower dose) likely play a role in the way the resistance to HDM201 takes place.

Collectively, our study has identified several potential mechanisms by which TP53 wild-type tumors may escape MDM2 targeted therapy, and provide evidence that Bcl-XL inhibition may be beneficial for relapsing patients that were treated with HDM201 intermittent scheduling.

Citation Format: Emilie A. Chapeau. Resistance mechanisms to TP53-MDM2 inhibition identified by in vivo piggyBac transposon mutagenesis screen [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5908.
Abstract 5922: "Gammora": Selective killing of cancer cells bearing CD24 by viral integrase
Authors: Shapira Shiran , Finkelshtein Eynat , Loyter Abraham , Arber Nadir
Published: August 2018

The HIV integrase is important in the integration of viral DNA to the host genome resulting in double-stranded breaks (DSB). Only 1-2 copies are integrated, due to inhibition of HIV-1 Rev protein, making the cells invisible to the immune system. CD24 is mostly expressed on cancer and not normal cells. We hypothesize that specific targeting of the viral particles using mAb to CD24 together with stimulating the enzyme activity and increasing the frequency of integration events, would increase the occurrence of DSB, that will lead to genomic instability, and ultimately to apoptosis of the target cells.

Aim

To develop a therapeutic platform, for selective eradication of cancer cells, which consists of targeted lentivirus particles and IN-derived peptides.

Methods

Integrase derived peptides (designated as INR2 and INS) were synthesized. Humanized anti-CD24 antibody (scFv) was engineered and fused to the lentivirus envelope. Stability, solubility and toxicity of the peptides were analyzed by HPLC-MS and MTD, respectively, as well as their activity and permeability into cells. Cell death was measured qualitatively by fluorescent microscopy and quantified by the enzymatic MTT assay. Colorectal, pancreatic and lung cancer cells were used for testing the potency of the lentiviral-based system. The efficacy of the suggested system was assessed in vivo, in nude mice bearing xenografts of lung cancer. The mice were treated systemically twice weekly.

Results

INS and INR2 stimulate viral integrase in vitro. A single IV administration of INR2 and INS was well tolerated without adverse effects. These lentivirus particles contain nonfunctional DNA with flanked LTRs allowing their integration into the target cells DNA and formation of DSB due to the action of the integrase whose activity was stimulated by the IN-derived peptides. Massive cell death (40-70%) in a dose-

dependent manner was achieved. Raltegravir, an antiretroviral FDA-approved drug, was able to inhibit cell death (2-3 times). In vivo, the INR2 and INS combination led to a significant reduction (50%) in tumor volume. Live imaging (IVIS device) of the tumors and selected organs confirmed that the viruses indeed reach the tumor after systemic injection and that the anti-CD24 enhanced the selectivity of infection.

Conclusions

The use of IN-derived peptides together with the CD24-targeted lenti-therapy approach suggests a novel strategy that specifically promotes killing and removal of a wide range of cancer cells.

Citation Format: Shiran Shapira, Eynat Finkelshtein, Abraham Loyter, Nadir Arber. "Gammora": Selective killing of cancer cells bearing CD24 by viral integrase [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5922.
Abstract 5928: Suicide gene therapy of ovarian cancer
Authors: Malekshah Obeid M. , Sarkar Siddik , Hatefi Arash
Published: August 2018

Purpose: The objective of this study was to develop a gene directed enzyme prodrug therapy (GDEPT) method that can effectively kill both differentiating and cancer stem-like cells (CSCs) in ovarian cancer.

Methods: Ovarian cancer cell lines including A2780, A2780-Cis, SKOV-3, OVCAR-3 and OVASC-1 (malignant cells from ascitic fluid of an ovarian cancer patient) were examined to identify the most drug resistant one. Various chemotherapeutic drugs which are used for suicide gene therapy were tested to identify one that is the most effective against ovarian cancer cells. These include 6-Methylpurine, 5-Fluorouracil, and SN-38. Cisplatin was used as a standard of care control. The anticancer activities of the drugs were studied by a cell toxicity assay and clonogenic assay. Flow cytometry was performed to study surface stem markers, such as CD133, CXCR4, HER2. ALDH1 activity in ovarian cancer cells was studied by ALDH assay kit. The ability of the most effective enzyme/prodrug system to kill drug resistant ovarian cancer cells was studied by a cell viability assay.

Results: The results of in vitro dose-response experiments on various ovarian cancer cells showed that SN-38 was the most efficient drug with IC50 in a range of 10 -100 nM, in comparison to 10-500 μM range of all other selected active drugs. OVASC-1 and OVCAR-3 cell lines showed a higher resistance to drug therapy as compared with other cell lines. It was observed that these two cell lines have a higher number of drug resistant cancer cells in their populations. It was also observed that CSCs are strongly associated with resistance to chemotherapy. ALDH assay, clonogenic assay and sphere formation assay showed a direct correlation between the therapy resistance and the percentage of CSCs. OVASC-1 cells in comparison to the other cell lines were found to be a suitable model for therapy resistant studies. After treatment by 100 nM SN-38, no visible colonies of Ascites were observed. Adipose-derived stem cells (ADSCs) were genetically modified to express Carboxylesterase-2 (CE2) and thus could convert CPT-11 to its active form (SN-38). Using this enzyme/prodrug system an effective eradication of highly resistant OVASC-1 ovarian cancer cells was demonstrated.

Conclusions: Our findings suggest that Carboxylesterase/CPT-11 enzyme/prodrug system is the most effective GDEPT method against highly drug resistant ovarian cancer cells.

Citation Format: Obeid M. Malekshah, Siddik Sarkar, Arash Hatefi. Suicide gene therapy of ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5928.
Abstract 603: Immune response to the influenza vaccination and heart rate variability in breast cancer survivors
Authors: Seiler Annina , LeRoy Angie S. , Fagundes Christopher P.
Published: August 2018

Background: Cancer treatment dysregulates immune function. These effects can persist long after the cessation of therapy and may diminish the immune system's ability to effectively mount an adequate immune response. Latent herpesvirus reactivation may mark a poor immune response to the influenza vaccination. Cancer treatment can also cause adverse effects on cardiac autonomic function as assessed by heart-rate variability (HRV), which is associated with higher morbidity and decreased survival in patients with cancer. Objective: To determine immune response to seasonal influenza vaccination as measured by human cytomegalovirus (CMV) IgG antibody titers and HRV in breast cancer survivors treated with radiation relative to healthy aged-matched controls. Methods: Serum samples for 13 breast cancer survivors within the first year of post-treatment and 13 healthy age-matched controls (mean age 44.7 years) were assayed for CMV IgG antibody titers and heart-rate variability was assessed using MindWare technologies. Group differences between cancer survivors versus healthy individuals were estimated using analysis of covariance (ANCOVA), with adjustment for age, body mass index (BMI) and race. Results: One-way ANCOVA modeling showed that breast cancer survivors treated with radiation had significantly higher antibody titers to CMV relative to age-matched healthy controls (F = 9.15 ; p <.01). While heart rate did not differ among the two groups (F = .36 ; p = .56), high frequency values were significantly lower in breast cancer survivors relative to the healthy controls (HF: F = 10.40 ; p <. 01). Conclusion: Breast cancer survivors manifested a dampened immune response to influenza vaccination and cardiovascular imbalance compared to healthy age-matched controls. These findings have important implications for long-term treatment outcomes.

Citation Format: Annina Seiler, Angie S. LeRoy, Christopher P. Fagundes. Immune response to the influenza vaccination and heart rate variability in breast cancer survivors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 603.
Abstract 612: Nivolumab-induced changes associated with patient outcomes are disparate in metastatic melanoma patient Tregs versus conventional T cells
Authors: Woods David M.
Published: August 2018

The PD1 blocking antibody nivolumab induces durable responses in metastatic melanoma patients. However, many patients do not respond and biomarkers differentiating patient response remain unidentified. To address this need, we investigated the effects of nivolumab on peripheral blood conventional CD4+ (Tcons) and regulatory T-cells (Tregs) of metastatic melanoma patients receiving nivolumab as adjuvant therapy or for metastatic disease. An increased ratio of PD1 expression on Tregs compared with Tcons was found at baseline patients with no evidence of disease (NED), while relapsing patients had a lower ratio (p<0.05). Intriguingly, NED patients had an increase in circulating Treg percentages post-nivolumab (p<0.05), which was not observed in relapsing patients. In vitro treatment of isolated T-cells with αPD1 resulted in increased proliferation (p<0.001). RNA-seq analysis of Tcons and Tregs showed distinct gene expression changes based on cell type and patient outcome. Pathway analysis demonstrated: upregulation of proliferation associated genes in NED Tregs, but not in relapse Tregs; downregulation of oxidative phosphorylation associated genes in NED patient Tregs and upregulation in NED Tcons; upregulation of heme metabolism associated genes in NED Tcons and downregulated in relapsing patient Tcons; and similar differences in MTORC1, heme metabolism and hypoxia pathways (all pathways q<0.01). Both Tcons and Tregs from non-relapsing patients had induction of phosphoSTAT3 (pSTAT3) expression post-nivolumab (p=0.012), which was absent in relapsing patients. This was also observed in a cohort of active disease patients (p=0.022), and the magnitude of change in pSTAT3 was positively correlated with patient survival (R2=0.49, p<0.05). Enhanced pSTAT3 expression was recapitulated in vitro in isolated T-cells (p<0.05). Post-nivolumab Tregs from patients with positive outcomes also had reduced suppressive function in mixed lymphocyte reactions (p<0.05), while patients with negative outcomes had no changes. In vitro treatment of Tregs with PD1 blocking antibodies resulted in a reduction in suppressive function (p<0.0001), while addition of a STAT3 inhibitor enhanced suppressive function (p<0.0001). These results suggest that nivolumab alters Treg signaling and metabolism pathways, hindering immune suppressive function. Collectively, our data demonstrate distinct effects of nivolumab on Tcons vs. Tregs, and highlight several potential biomarkers of patient benefit, warranting further exploration.

Citation Format: David M. Woods. Nivolumab-induced changes associated with patient outcomes are disparate in metastatic melanoma patient Tregs versus conventional T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 612.
Abstract 631: Intratumoral activation of STING with a synthetic cyclic dinucleotide elicits antitumor CD8 T-cell immunity that effectively combines with checkpoint inhibitors
Authors: Desbien Anthony L. , Gauthier Kelsey Sivick , Corrales Leticia , Reiner Gabrielle , Glickman Laura Hix , Katibah George , Hudson Thomas E. , Vu Uyen , Surh Natalie H. , Francica Brian , Deng Weiwen , Kanne David B. , Leong Justin J. , Ndubaku Chudi , Metchette Ken , McKenna Jeffery M. , Bender Steven L. , Leong Meredith L. , Dubensky Thomas W. , Elsas Andrea van , McWhirter Sarah M.
Published: August 2018

Activation of the STING pathway by intratumoral (IT) injection of synthetic cyclic dinucleotides (CDNs) induces stable tumor regression in preclinical models, yet the underlying immune correlates are not fully understood. ADU-S100, a CDN under clinical evaluation, was administered IT with an optimized dosing regimen to explore the immune requirements for antitumor efficacy in mouse syngeneic tumor models. We show that CD8+ T cells are necessary and sufficient for durable antitumor immunity elicited by ADU-S100 and that activation of STING in hematopoietic cells mediates CD8+ T cell induction. Both type I IFN and TNFα, which are induced by STING pathway activation, influence the antitumor immune response. The combination of ADU-S100 and anti-PD1 treatment enhances CD8+ T cell-dependent, noninjected tumor control that correlates with an enhanced effector profile of CD8+ T cells in the tumor. Combination of ADU-S100 with checkpoint inhibition also enhances durable immunity in a poorly immunogenic tumor model. Together, these results elucidate the immune correlates to STING-mediated antitumor efficacy and highlight the potential of combining STING agonists with checkpoint inhibition in the clinic.

Citation Format: Anthony L. Desbien, Kelsey Sivick Gauthier, Leticia Corrales, Gabrielle Reiner, Laura Hix Glickman, George Katibah, Thomas E. Hudson, Uyen Vu, Natalie H. Surh, Brian Francica, Weiwen Deng, David B. Kanne, Justin J. Leong, Chudi Ndubaku, Ken Metchette, Jeffery M. McKenna, Steven L. Bender, Meredith L. Leong, Thomas W. Dubensky Jr., Andrea van Elsas, Sarah M. McWhirter. Intratumoral activation of STING with a synthetic cyclic dinucleotide elicits antitumor CD8 T-cell immunity that effectively combines with checkpoint inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 631.
Abstract 635: Characterizing PD-L1 expression in tumor-associated cells and CTC using IncellDx and Celsee molecular diagnostic technologies
Authors: Fazeli Fariba , Sharma Vishal Premdev , Chow William Chiro , Chargin Amanda , Gogoi Priya , Handique Kalyan , Patterson Bruce
Published: August 2018

Detection of PD-L1 expression in NSCLC cancer patients is increasingly becoming a widely used molecular diagnostic tool. Evidence also suggests that PD-L1 can be induced with radiotherapy and may be an immune escape mechanism in cancer. Current methods analyze PD-L1 expression in either solid tumor biopsies or circulating tumor cells (CTC), independent of each other. In order to design a comprehensive PD-L1 treatment regime, it is imperative to look at the PD-L1 expression in cancer cells of the tumor as well as in CTCs. In this study we present a comprehensive profiling of PD-L1 expression in solid tumor biopsy and CTC of 10 lung cancer patients. We further extend the study by evaluating the co-relation of the PD-L1 expression with other immune cell infiltration markers in the lung biopsy samples. There is no single technology that can provide a comprehensive profiling of PD-L1 expression in lung biopsies as well as the CTCs isolated from the peripheral blood. Using two very different and unique molecular diagnostic technologies, IncellDx OncoTect iO Single Cell Quantitative PD-L1 assay and automated CTC analysis Celsee C-PREP GENESIS, we present the first report discussing the PD-L1 expression in tumor cells of the solid and liquid biopsies and its correlation with DNA content and infiltrating immune cell markers. Nine out of the ten patient samples evaluated showed strong correlation between DNA content and PD-L1 expression in tumor-associated cells and CTC. One aneuploid patient sample showed very low expression of PD-L1 in tumor-associated cancer cells; however, it still showed a positive staining for PD-L1 in CTC. Obtaining a comprehensive PD-L1 expression and associated immune cell marker profile in solid tumor biopsy and circulating tumor cells may be needed to design a more effective and lasting immunotherapy to treat NSCLC lung cancer patients. The results discussed in this study will strongly impact the current understanding and implementation of the molecular diagnostics tools to evaluate cancer patients in order to define a robust therapeutic regimen.

Citation Format: Fariba Fazeli, Vishal Premdev Sharma, William Chiro Chow, Amanda Chargin, Priya Gogoi, Kalyan Handique, Bruce Patterson. Characterizing PD-L1 expression in tumor-associated cells and CTC using IncellDx and Celsee molecular diagnostic technologies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 635.
Abstract 638: Microbubble enrichment of CTCs
Authors: Jones Thomas , Ostruszka Leo J. , Fouladdel Shamileh , Azizi Ebrahim , Slamecka Jaroslav , McNaughton Brandon H. , Younger John , Wicha Max , McClellan Steven
Published: August 2018

Despite numerous new cell analysis technologies, contaminating red- and white blood cells remain a central barrier to the efficient characterization of circulating tumor cells. Microbubbles, which capture and separate cells by flotation, recently have been described as an alternative or adjunct to magnetic beads or microfluidic methods for CTC sample preparation. This technology may find particular value in applications where mechanical force needs to be minimized or magnetization of cells needs to be avoided. Here, we evaluate novel RBC-targeting microbubbles for buoyancy-based enrichment of CTCs. Cultured human pancreatic cancer cells (line CFPAC-1, ATCC CRL 1918) were spiked into standardized samples representative of common CTC workflows (106 RBC : 5x104 WBC : 1.5x102 tumor cells). Microbubbles bearing antibodies to human erythrocytes were added at various Bubble:RBC ratios and mixed via trituration for 1 minute. Samples were then centrifuged, the microbubble-captured RBCs removed, and the remaining pelleted cells analyzed for the fraction of RBCs removed and the extent of off-target depletion of tumor cells. Cells were quantified with an automated cell counter, and all experimental conditions were performed in at least triplicate. RBC removal rose with increasing Bubble:RBC ratio ( p < 0.001 ). At a ratio of 1:1, 92 +/- 1% of RBCs were removed with one minute of mixing; at a ratio of 1.5:1, 97 +/- 1% of RBCs were removed. Off-target binding of CFPAC-1 cells was minimal at a Bubble:RBC ratio of 1:1, with 0 +/- 6% of tumor cells captured. Off-target capture increased at a Bubble:RBC ratio of 2:1, with 12 +/- 9% tumor cell loss from the experimental samples. Using a standardized blood sample with a clinically relevant low concentration of cultured tumor cells, microbubbles targeting RBCs and dosed at a ratio of one bubble per target erythrocyte were able to achieve > 90% removal without statistically detectable loss of a cultured cancer line. Higher bubble concentrations resulted in more exhaustive RBC removal, but with some loss of cultured cells. Microbubble-based cell separation may have a useful role in CTC purification protocols.

Citation Format: Thomas Jones, Leo J. Ostruszka, Shamileh Fouladdel, Ebrahim Azizi, Jaroslav Slamecka, Brandon H. McNaughton, John Younger, Max Wicha, Steven McClellan. Microbubble enrichment of CTCs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 638.
Abstract 643: An ultra-sensitive multiplex allele-specific real-time PCR (Udx-PCR) assay for detection of KRAS/BRAF/NRAS mutations in colorectal cancer
Authors: Zhai Jianxin , Wu Youmin , Luo Xuan , Li Xinsheng , Yu De-Hua
Published: August 2018

RAS and BRAF gene mutation assessment is essential for therapy decision in metastatic colorectal cancer patients with anti-EGFR monoclonal antibody treatment. However the currently approved assays in clinic is mainly focusing on tissue samples with detecting sensitivity lower than 1% in 10ng genomic DNA background. The emerging analysis of ctDNA offers advantages in clinical diagnosis as a feasible method, which requires much more sensitive methodology due to low abundance in plasma samples. Herein, we developed a multiplex allele-specific real-time (ARMS) PCR assay for detection fourteen mutations of KRAS/BRAF/NRAS in an 8-tube PCR strip. The reagent and format of mutation detection were particularly optimized to reach high specificity and sensitivity. Performance evaluation showed that the assay is remarkably sensitive with lower detection limit of 0.1% in a background of 10ng wild-type genomic DNA. A total of 136 CRC FFPE samples were screened by using the assay. Sixty-two samples were positive for KRAS/BRAF/NRAS mutations, and fifty-eight were confirmed by the Sanger sequencing. The four discrepant samples were tested further by ddPCR and the results revealed that the allelic frequencies in these samples were as low as 0.1% to 5%. The evaluation of ctDNA detection is being carried in metastatic colorectal cancer patients. The multiplex allele-specific real-time PCR assay provides a rapid, reliable and low-cost diagnostic tool for accurate detection of KRAS/BRAF/NRAS mutations for clinical application.

Citation Format: Jianxin Zhai, Youmin Wu, Xuan Luo, Xinsheng Li, De-Hua Yu. An ultra-sensitive multiplex allele-specific real-time PCR (Udx-PCR) assay for detection of KRAS/BRAF/NRAS mutations in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 643.
Abstract 666: Gut microbiota SCFA modulates DCs antigen presentation and impacts tumor response to radiotherapy
Authors: Facciabene Andrea , Rafail Stavros , Gomez Luis Gil de , Pierini Stefano , Uribe-Herranz Mireia , Bittinger Kyle
Published: August 2018

Alterations in gut microbiota modulate host physiologic functions, including immune responses, and they play a role in the pathophysiology of several diseases, including cancer. Radiotherapy (RT), an established curative and palliative cancer treatment, exerts potent immune modulatory effects, inducing tumor-associated antigen (TAA) cross-priming with antitumor CD8+ T cell elicitation and abscopal effects. Herein, we tested whether the gut microbiota modulates antitumor immune response following RT. Vancomycin, an antibiotic that acts mainly on gram-positive bacteria and is restricted to the gut, potentiated the RT-induced antitumor immune response and tumor growth inhibition. This synergy was dependent on tumor-associated antigen cross-presentation enanchement, cytolytic CD8+ T cells and on IFN-g. Notably, butyrate, a metabolite produced by the vancomycin-depleted gut bacteria, abrogated the vancomycin effect. In conclusion, gram-positive bacteria depletion by vancomycin enhances the antitumor activity of RT, which has important clinical ramifications.

Citation Format: Andrea Facciabene, Stavros Rafail, Luis Gil de Gomez, Stefano Pierini, Mireia Uribe-Herranz, Kyle Bittinger. Gut microbiota SCFA modulates DCs antigen presentation and impacts tumor response to radiotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 666.
Abstract 669: The regulation of ALOX15 expression in colorectal cancer to overcome the resistance to radiation therapy
Authors: Na Yoo Jin , Lee Dae-Hee , Kim Bo Ram , Kim Jung Lim , Jeong So Yeon , Park Seong Hye , Jo Min Jee , Jeong Yoon A , Yun Hye Kyeong , Oh Sang Cheul
Published: August 2018

Colorectal cancer (CRC) is the third most common cancer and the third leading cause of 'cancer related death' world-wide. In case of CRC patients who have large sized tumor, radiation therapy is essential to decrease tumor size without having to eliminate the anus. However, radiation effects vary according to patients. Also, the association between genes and radiation sensitivity is still rarely researched. In this study, we elucidated that the expression level of ALOX15 regulates radiation sensitivity by TNFα signaling pathway in CRC cells. ALOX15 is known as the main metabolic enzyme for linoleic acid and arachidonic acid. Interestingly, stable cells which express low ALOX15 inhibit radiation-induced apoptosis. Mechanistically, inhibition of ALOX15 increases NF-kB under radiation therapy condition. NF-kB that plays a key role in the cellular response to DNA damage is a reason for ALOX15 to act as a radiation sensitizer. Taken together, our results indicate that ALOX15 is suggested as a prediction marker for radiation therapy. In addition, screening natural compound for elevating ALOX15 would be used for the purpose of combination therapy in radiation-resistant CRC cells.

Citation Format: Yoo Jin Na, Dae-Hee Lee, Bo Ram Kim, Jung Lim Kim, So Yeon Jeong, Seong Hye Park, Min Jee Jo, Yoon A Jeong, Hye Kyeong Yun, Sang Cheul Oh. The regulation of ALOX15 expression in colorectal cancer to overcome the resistance to radiation therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 669.
Abstract 68: Targeting macrophage that forms a chemotherapy-induced niche significantly reduces cancer stem cells and tumors
Authors: He Xi , Giles Paloma I. , Lin Edward H. , Li Linheng
Published: August 2018

Failure in cancer therapy is often caused by relapse resulting largely from chemo-resistant cancer stem cells (CSCs). Most colorectal cancer patients, after receiving chemoradiotherapy, still harbor treatment-resistant residual tumor cells that are akin to colorectal adenoma. We therefore exploited the APCMin adenoma model that allowed us to study the natural and dynamic interaction between CSCs and their microenvironment in vivo following therapeutic stress. We observed that macrophages were directly recruited from blood vessel to the CSC niche in response to chemotherapy, playing a role in protecting and further promoting activation of CSCs. Reduction in the Ly6c-CD11bintCX3CR1+ MHCII+ population of tumor associated macrophages (TAMs) by celecoxib or clodrosome was correlated with a decline in slow-cycling drug-resistant CSCs and tumorigenesis. Molecularly, we found that COX-2 inhibitor celecoxib suppressed Akt and Wnt signaling, downstream of macrophage-dependent PGE2-EP pathway, thus reducing CSC survival and activation. Taken together, in the naturally formed adenoma reflecting human familial adenomatous polyposis, Chemotherapy-induced recruitment of TAMs to the CSC niche protected and promoted activation of surviving CSCs via PGE2-EP mediated Akt-Wnt signaling, leading to tumor regrowth. Targeting the TAMs will benefit clinical treatment of colorectal cancer.

Citation Format: Xi He, Paloma I. Giles, Edward H. Lin, Linheng Li. Targeting macrophage that forms a chemotherapy-induced niche significantly reduces cancer stem cells and tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 68.