bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1618: Simultaneous isolation of exosomes and cfDNA from liquid biopsies using universal kit based on SubX-MatrixTM technology
Authors: Malykh Andrei G. , Malek Anastasia , Lokshin Anna , Evtushenko Vladimir 
Published: August 2018

Abstract
In 2016 we have developed novel approach for isolation of circulating cell-free DNA employing the proprietary bi-functional substance SubX® that tightly binds DNA and forms sandwich [cfDNA- SubX®-magnetic particles]. These sandwich aggregates are easily separated from bio-liquid with magnet followed by cfDNA isolation by speedy protocol. Our approach does not require Proteinase K digestion and 2-3-fold dilution of bio-liquid with lysis/binding buffer in contrast to all commercially available kits. Our kit has been evaluated in various research and clinical labs for cfDNA isolation from serum, plasma and urine of patients with different forms of cancer. SubX®-approach also allows for isolation of cfDNA from large volumes of urine samples without its concentration. We demonstrated linear increase of the DNA yield with liquid biopsy volume. Another unique property of the SubX® is that both ends of the molecule display lipid binding groups. This feature allows each molecule of SubX® to anchor two exosomes (i.e. dimerize). Excess of SubX® molecules in the solution results in aggregation of up to 10-15 exosomes and formation of micron-size particles that are easily precipitated in a brief 14K x g (or 30 min 2K x g) centrifugation step. A specially designed buffer allows for reconstitution of the pelleted exosomes back to monomer format for downstream applications. Flow cytometry with specific surface marker CD9 confirmed that SubX®-aggregated vesicles are exosomes. Analysis of membrane-bound protein markers and microRNA profile of exosomes isolated from plasma of prostate cancer patients will be presented. Difference in SubX® binding strength for cfDNA and exosomes allows for consequent isolation from the [cfDNA/exosomes- SubX®-magnetic particles] pellet first exosomes, then cfDNA. Eventually, we combined both approaches in one kit for isolation of cfDNA and exosomes from the same sample. This simultaneous isolation protocol is simple and fast, it allows to employ minimal volume of valuable bio samples as well as to reduce labor and cost. The kit was validated in serum and urine samples from patients with ovarian and pancreatic cancer. Exosome nature of isolated vesicles was confirmed by three independents methods - light scattering, immunological and electron microscopy. Thus, one could analyze both genetic (DNA and RNA) as well as protein markers present in the same liquid biopsy sample.
Citation Format: Andrei G. Malykh, Anastasia Malek, Anna Lokshin, Vladimir Evtushenko. Simultaneous isolation of exosomes and cfDNA from liquid biopsies using universal kit based on SubX-MatrixTM technology [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 1618.
Abstract 1634: BRCA-like phenotype constitutes hallmark of osteosarcoma
Authors: Kovac Michal , Ribi Sebastian , Blattmann Claudia , Nathrath Michaela , Baumhoer Daniel 
Published: August 2018

Abstract
Background: Osteosarcomas are aggressive bone tumours with a high degree of genetic heterogeneity, which limits treatment options in patients who do not respond to standard chemotherapy.
Methods: We sequenced 104 exomes and low-coverage genomes from 14 time-series consisting of chemotherapy-naïve biopsies, resection specimens, locally recurring tumours and/or distant metastases. We then identified cancer driver mutations, signatures of mutation processes and reconciled clonal structures of tumours with phylogenetic trees, thus building and evolutionary history of each tumour. Where possible, we searched for the evidence of selection acting on the tumours and molecular hallmarks that could be therapeutically exploited.
Results: Interrogating genomic data from time series, we identified pathogenic mutations in at least 10 cancer genes, but there was no universal cancer driver event present in the majority of tumours at any given time point. However, &gt;80% of osteosarcomas exhibited specific combination of single-base substitutions, LOH and large-scale genome instability characteristic of BRCA1/2-deficient tumours. In osteosarcoma, "BRCAness" developed recurrently through independent BRCA1/2 mutations and mutations in their binding partners such as PALB2 or CHEK2. We therefore investigated the sensitivity of a panel of osteosarcoma cell lines to the poly(ADP)-ribose polymerase inhibitor talazoparib. In combination with temozolomide, talazoparib led to the to good response of osteosarcoma cell lines, activation of BAX and BAK, loss of mitochondrial membrane potential, DNA fragmentation and caspase-dependent cell death.
Conclusion: The acquisition of a BRCA-like phenotype in osteosarcoma has implications for the development of novel treatment strategies using PARP inhibitors alone, or together with standard chemotherapy.
Citation Format: Michal Kovac, Sebastian Ribi, Claudia Blattmann, Michaela Nathrath, Daniel Baumhoer. BRCA-like phenotype constitutes hallmark of osteosarcoma [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 1634.
Abstract 1639: Androgen receptor and ALDH1 expression among internationally diverse patient populations
Authors: Jiagge Evelyn M. , Jibri Aisha , Bensenhaver Jessica , Wicha Max , Awuah Baffour , Merajver Sofia , Newman Lisa 
Published: August 2018

Abstract
Background: Population-based incidence rates of breast cancers that are negative for the estrogen receptor (ER), progesterone receptor (PR), HER2/neu triple-negative breast cancer (TNBC) are higher among African American (AA) compared to White American (WA) women, and several studies suggest that TNBC prevalence is increased among selected populations of African patients. The colonial-era transatlantic slave trade resulted in shared ancestry between contemporary AA and Gh populations. The extent to which associations between TNBC are related to East African versus West African ancestry, and whether these associations extend to expression of additional hormone receptors such as androgen receptor (AR) and stem cell markers such as ALDH1, is uncertain, but this research may explain breast cancer disparities between domestic communities within the United States as well as between international population subsets.
Methods: We utilized immunohistochemistry to evaluate ER, PR, HER2/neu, AR and ALDH1 expression among White American (n=153), African American (n=76), Ethiopian (Eth)/East African (n=90), and Ghanaian (Gh)/West African (n=286) breast cancers through an IRB-approved international research program.
Results: Mean age at breast cancer diagnosis was 43, 49, 60, and 57 years for the Eth, Gh, AA, and WA patients, respectively. Frequency of TNBC was significantly higher for the AA and Gh patients (41% and 54%, respectively) compared to the WA and Eth patients (23% and 15%, respectively; p&lt;0.001). These associations were unchanged when limited to patients age 50 years and younger (47% and 49% for AA and Gh, respectively, versus 18% and 16% for WA and Eth, respectively; p&lt;0.001). Frequency of ALDH1 positivity was also higher for the AA and Gh tumors (32% and 36%, respectively) compared to the WA and Eth tumors (23% and 17%, respectively; p=0.007). Significant differences were also observed for distribution of AR positivity, which was 71%, 55%, 42%, and 50% for the WA, AA, Gh, and Eth cases, respectively (p=0.008). Frequency of ALDH1 expression was numerically higher in the AA, Gh, and Eth TNBC cases compared to the WA TNBC cases, but this was not statistically significant (33%, 41%, and 31%, respectively, versus 18%; p=0.47).
Conclusions: Extent of African ancestry appears to be associated with particular breast cancer phenotypes. West African ancestry correlates with increased risk of TNBC and breast cancers that are positive for ALDH1.
Citation Format: Evelyn M. Jiagge, Aisha Jibri, Jessica Bensenhaver, Max Wicha, Baffour Awuah, Sofia Merajver, Lisa Newman. Androgen receptor and ALDH1 expression among internationally diverse patient populations [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 1639.
Abstract 1641: Utilization of palliative therapies among Hispanics with stage IV non-small cell lung cancer
Authors: Duma Narjust , Durani Urshila , Molina Julian , Moynihan Timothy J. 
Published: August 2018

Abstract
Background: Racial disparities in the treatment of lung cancer are well documented. However, research in disparities in palliative care is limited. Early integration of palliative care in advanced non-small cell lung cancer (NSCLC) has been proven to improve quality of life and overall survival in this subset of patients. We proposed to study the use of palliative treatments for stage IV NSCLC among Hispanic patients (pts). Methods: Using the National Cancer Database (NCDB), we identified all Hispanic pts (self-reported) diagnosed with stage IV NSCLC from 2005 to 2013. Cases with incomplete data, unknown life/death status or classified as Hispanic by surname only were excluded. Hispanics were grouped based on place of origin. Pearson chi-square tests were used to estimate differences in categorical data; predictors of palliative care referral/use were determined by logistic regression analysis. Results: 10,441 pts were included. The median age was 66 years. Regarding place of origin: 15.5% of pts were from Mexico, 8% from South/Central America, 7% from Cuba, 6% from Puerto Rico, 2.4% from the Dominican Republic and 57.6% were no-otherwise specified. When all Hispanic pts were included, 3.5% received surgery, 45% radiation and 52.9% chemotherapy. Overall, 2.2% of pts received a referral for palliative pain management +/- other palliative therapies. When divided by place of origin, Dominican pts had the highest percentage of pain management referrals at 5.5% (p&lt;0.01) followed by Puerto Rican patients (2.4%). On the other hand, only 1.3% of pts from South/Central America were referred to pain management. In multivariate analysis, Dominican Republic origin (OR: 3.30, 95%CI: 1.69-6.44, p&lt;0.01), bone metastasis (OR: 1.98, 95%CI: 1.17-3.3, p&lt;0.01) and a Charlson comorbidity index ≥2 (OR: 2.07, 95%CI: 1.11-3.85, p&lt;0.02) were significant predictors of receiving a pain management referral. We observed an increased number of pain management referrals over time with 1.4% of Hispanic patients getting a referral in 2004 vs. 2.9% in 2013 (p&lt;0.02). Conclusions: We observed that only a small percentage of Hispanic pts with metastatic NSCLC cancer are receiving referral for palliative care/pain management. Several cultural beliefs and barriers may play a role in these findings. Providers should offer early referrals to pain management/palliative care to all patients with metastatic NSCLC independent of their race or ethnicity.
Citation Format: Narjust Duma, Urshila Durani, Julian Molina, Timothy J. Moynihan. Utilization of palliative therapies among Hispanics with stage IV non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1641.
Abstract 1707: Tumor treating fields exert cellular and immunologic effects
Authors: Wong Eric T. , Timmons Joshua , Swanson Kenneth D. 
Published: August 2018

Abstract
While there is clear clinical evidence to support the therapeutic efficacy of low dose alternating electric fields (TTFields) in treating glioblastoma, their intracellular targets and mechanism of action remains poorly defined. Historically, TTFields have been thought to operate by inducing extensive plasma membrane blebbing during the entry into anaphase resulting in aberrant mitotic exit and cell death. However, we have found that cells exposed to TTFields exhibited increased granularity and evidence of endoplasmic reticulum stress and genotoxic stress suggesting other mechanisms to explain their clinical efficacy. Cells exposed to TTFields exhibited reductions in levels of culture media acidification and analysis of intracellular metabolism showed evidence for an increase in mitochondrial respiration and glutaminolysis pathway. Our previous analysis of data from the pivotal phase III EF-11 clinical trial showed that increased corticosteroid use was strongly correlated with poor outcomes in TTFields-treated patients, possibly by suppressing immune effector function required for to drive tumor regression. TTFields drives the cell surface expression of immune stimulatory stress marker proteins calreticulin, and the secretion of the Alarmin/DAMP protein, HMGB1 which are together suggest that TTFields can stimulate immunogenic cell death. Co-culture of bone marrow derived macrophages with TTFields-treated CT26 cells resulted in the up-regulation of cell surface activation markers in the macrophage population that was blocked by a monoclonal antibody directed against HMGB1. Further, supernatants from these co-cultures contained increased levels of pro-inflammatory cytokines and chemokines and a reduction of anti-inflammatory cytokines associated with the presence of the TTFields-treated cells relative to co-cultures containing naïve tumor cells. Further, in vivo, engrafted Lewis lung carcinoma tumors exhibited increased levels of granzyme positive cells following TTFields treatment. Together, these data suggest that TTFields affect cancer cells during interphase and this insult leads to alterations in cancer cell metabolism and increased tumor cell immunogenicity and thus act to drive increased immune recognition and tumor rejection. In patients, these effects are likely to be opposed by the immune suppressive functions within the tumor microenvironment and corticosteroid use. Therefore, this model of TTFields action may provide a therapeutically rational base to devise improved TTFields treatment strategies from.
Citation Format: Eric T. Wong, Joshua Timmons, Kenneth D. Swanson. Tumor treating fields exert cellular and immunologic effects [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 1707.
Abstract 1710: DKN-01, a therapeutic DKK1 neutralizing antibody, has immune modulatory activity in nonclinical tumor models
Authors: Haas Michael , Heath Heidi , Wu Yinyuan , Zhang Xinjun , Sirard Cynthia , He Xi , Newman Walter , Kagey Michael 
Published: August 2018

Abstract
Wnt signaling is a fundamental pathway that is dysregulated in oncology. The Wnt signaling modulator DKK1 is expressed in a variety of tumor types and elevated levels frequently correlate with poor survival. DKK1 promotes tumor growth by stimulating proliferation, metastasis, and angiogenesis, and has been implicated in contributing to an immune suppressive tumor microenvironment. DKN-01 is a humanized monoclonal therapeutic antibody that binds DKK1 with high affinity and selectivity. It is currently being evaluated clinically as a monotherapy and in combination in a variety of solid tumors. Here we describe further characterization of the mechanism of action of DKN-01 and demonstrate immune mediated anti-tumor activity in nonclinical models. A murine version of DKN-01 (mDKN-01) has efficacy in a syngeneic melanoma B16 tumor model. However, mDKN-01 is unable to impede B16 tumor growth in NSG immunodeficient mice, indicating that a functioning immune system is required for antibody activity. Furthermore, preliminary data suggest that mDKN-01 is targeting a myeloid derived suppressor cell population in the tumor microenvironment. These data support an immune mediated mechanism of action of DKN-01 and provide a rationale for clinical development in combination with immunotherapy agents. The first clinical study evaluating DKN-01 in combination with pembrolizumab has initiated enrollment in patients with relapse/refractory esophagogastric malignancies.
Citation Format: Michael Haas, Heidi Heath, Yinyuan Wu, Xinjun Zhang, Cynthia Sirard, Xi He, Walter Newman, Michael Kagey. DKN-01, a therapeutic DKK1 neutralizing antibody, has immune modulatory activity in nonclinical tumor models [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 1710.
Abstract 1728: Nonalcoholic steatohepatitis (NASH) impairs treatment of intrahepatic metastases with CD4+ T cell dependent RNA vaccine
Authors: Heinrich Bernd , Brown Zachary J. , Mathias Vormehr , Fu Qiong , Ma Chi , Yu Su Jong , Sahin Ugur , Greten Tim F. 
Published: August 2018

Abstract
Nonalcoholic fatty liver disease (NAFLD) is the most common liver disorder in Western industrialized countries. NAFLD along with its advanced form non-alcoholic steatohepatitis (NASH) are becoming an increasing risk factor for the development of cirrhosis and subsequently for hepatocellular carcinoma (HCC). We showed a selective CD4+ T cell loss in mice fed with methionine-choline deficient (MCD) diet, a model to induce NASH (Ma et al. Nature 2016). This selective loss of CD4 T cells may not only lead to immune escape by tumor cells but also lessen the effectiveness of immunotherapy. Kreiter et al. created a RNA vaccine from B16 melanoma cells encoding the neo-epitope M30, which is presented by MHC class II and recognized by CD4+ T cells. The vaccine showed therapeutic immune response in mouse models of subcutaneous B16-tumors and lung metastases (Kreiter et al. Nature 2015). Here we present an in vivo study of M30 RNA vaccine in a mouse model of B16 melanoma liver metastasis induced by intrahepatic injection. Mice were separated into groups and fed regular chow or MCD diet. B16F10-GFP-Luc melanoma cells were intrahepatically injected and mice were treated with M30 RNA or irrelevant RNA as control via tail vein injection. Intrahepatic tumor growth was monitored by bioluminescence imaging over the course of treatment. Mice were sacrificed and immunomonitoring of tumor surrounding liver tissue and tumor infiltrating lymphocytes (TIL) was performed. Mice treated with M30 RNA vaccine showed reduced tumor growth of intrahepatic metastasis compared to irrelevant RNA. Mice fed MCD diet could not control tumor growth either receiving M30 RNA vaccine treatment or irrelevant RNA. This impaired function of the treatment was related to a loss of CD4+ T cells in the NASH liver and a more immunosuppressive immune cell phenotype in the tumor environment.
This study indicates impaired efficacy of cancer immunotherapy due to fatty liver disease and subsequently changed immune response due to loss of CD4+ T cells as the main effector immune cell. As the M30 RNA vaccine reduced tumor growth in non-fatty liver, this data might be considered when planning future immunotherapy treatment strategies for comorbid patients with fatty liver disease as CD4+ T cell based treatment strategies might be impaired in patients with NASH.
Citation Format: Bernd Heinrich, Zachary J. Brown, Vormehr Mathias, Qiong Fu, Chi Ma, Su Jong Yu, Ugur Sahin, Tim F. Greten. Nonalcoholic steatohepatitis (NASH) impairs treatment of intrahepatic metastases with CD4+ T cell dependent RNA vaccine [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 1728.
Abstract 1733: Wnt antagonists synergize with immune checkpoint inhibitors to enhance anti-tumor responses
Authors: Ross Jenny , Hoey Timothy , Kapoun Ann , Lewicki John , Gurney Austin , Murriel Christopher L. 
Published: August 2018

Abstract
Aberrant Wnt signaling through overexpression or activating mutations in Wnt pathway proteins is responsible for the initiation and progression of numerous cancers. While enhanced Wnt signaling has been shown to play a major role in cancer stem cell biology, more recent studies have implicated Wnt in the development of resistance to anti-tumor immune responses. Here, we show that in various syngeneic murine tumor models, targeting Wnt signaling using the Fzd receptor monoclonal antagonist antibody, OMP-18R5 (vantictumab) or the pan-Wnt decoy receptor Fc fusion protein OMP-Fzd8-Fc (ipafricept) in combination with immune checkpoint inhibitors anti-CTLA-4 or anti-PD1 induce synergistic anti-tumor responses leading to decreased tumor volume and increased infiltration of activated CD8+ T cells into the tumor microenvironment. In addition, we show that combined Wnt and immune checkpoint inhibition decrease regulatory T cells (Tregs), enhance cytotoxic T cell activity and increase antigen presentation by APCs. More importantly, we demonstrate that anti-PD1 and the Wnt antagonists decrease immune suppressive myeloid cell populations, which may enhance therapeutic efficacy and anti-tumor responses. Therefore, these results suggest that co-targeting Wnt and immune checkpoint proteins may provide valuable opportunities for novel combination strategies for immunotherapeutic clinical development.
Citation Format: Jenny Ross, Timothy Hoey, Ann Kapoun, John Lewicki, Austin Gurney, Christopher L. Murriel. Wnt antagonists synergize with immune checkpoint inhibitors to enhance anti-tumor responses [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1733.