tbuckley/cabot-search huggingface dataset:
Ultraviolet B Irradiation Causes Stimulator of Interferon Genes–Dependent Production of Protective Type I Interferon in Mouse Skin by Recruited Inflammatory Monocytes
Authors: Clayton Sontheimer, Denny Liggitt, Keith B. Elkon
Journal: Arthritis & Rheumatology (2016)

Objective Photosensitivity is common in patients with systemic lupus erythematosus, although the mechanisms linking ultraviolet (UV) light to flares are not well understood. We undertook this study to determine whether repetitive UVB exposure could induce type I interferon (IFN) production in normal mouse skin, and to investigate the roles of inflammatory monocytes and plasmacytoid dendritic cells (PDCs) in type I IFN production and development of UVB irradiation–induced inflammation. Methods Mice were irradiated with UVB at 100 mJ/cm 2 for 5 days, and cutaneous manifestations were examined by messenger RNA expression of inflammatory and type I IFN response genes, histology, and flow cytometry. Inflammatory monocyte and PDC depletion experiments were performed in CCR2–diphtheria toxin receptor (DTR)–transgenic mice and blood dendritic cell antigen 2–DTR–transgenic mice. The roles of type I IFN and of the adaptor protein stimulator of IFN genes (STING) in UVB irradiation–induced inflammation were investigated using IFN‐α/β/ω receptor (IFNAR)–knockout mice and STING‐knockout mice. Results Repeated UVB irradiation stimulated an inflammatory cell infiltrate and induction of type I IFN and proinflammatory cytokines. Interestingly, the type I IFN response was independent of PDCs but dependent on inflammatory monocytes, which were recruited following UVB irradiation. The adaptor protein STING was necessary for both type I IFN and proinflammatory cytokine expression in the skin. UVB‐irradiated IFNAR‐knockout mice showed increased levels of proinflammatory genes and more severe inflammation by histology, suggesting a protective role for type I IFN. Conclusion In wild‐type mice, repeated doses of UVB irradiation induce monocyte‐dependent and PDC‐independent expression of type I IFN together with expression of other proinflammatory cytokines. Induction is dependent on the adaptor protein STING. Surprisingly, studies using IFNAR‐deficient mice revealed that type I IFN protects against UVB irradiation–induced skin inflammation, in part by attenuating proinflammatory cytokine expression and limiting tissue damage.
Abstract 283: Establishing the predominant type of leukemia among patients attending the Uganda Cancer Institute
Authors: Gerald Nsubuga
Journal: Cancer Research (2017)

Abstract Introduction: Leukemia is a cancer of white blood cells. It is characterized by an abnormal accumulation of WBC in the bone marrow which leads to bone marrow failure and is manifested in form of anemia and thrombocytopenia. This arises from neoplastic transformation usually of a single cell. There are four common types of leukemia: 1) Acute lymphoblastic leukemia (ALL); 2) Acute myeloblastic leukemia (AML); 3) Chronic lymphocytic leukemia (CLL); and Chronic myeloid/myclocytic leukemia (CML). Methods: The study was designed to find out the predominant type of leukemia. 80 leukemia cases which comprised of all bone marrow aspirates and peripheral blood slide results with specific age, sex and ward of the patients was analyzed in the hematology laboratory. Results: The study was revealed that leukemia occurred in following order of types of predominant that is AML 15 (18.75%), CLL 12(15%) and least type was CML 11(13.75%). The result also showed that the most affected age group by all age. Brackets of (01-20) years. From the study a male preponderance (M: F) of 4:3 was found in cases of ALL. The study emphasizes that special staining techniques be adopted in diagnosing and typing of leukemia. Conclusions: From the results presented, ALL is the predominant type of leukemia affecting children and young adults. AML was found to affect children, young adults, and adults. CLL and CML were found to occur at the same incidence, the elderly (61-70 and 71-80 years). Other types were rare in these age group similarly CLL and CML were rare in children and young adults. Note: This abstract was not presented at the meeting. Citation Format: Gerald Nsubuga. Establishing the predominant type of leukemia among patients attending the Uganda Cancer Institute [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 283. doi:10.1158/1538-7445.AM2017-283
Abstract 3802: A novel uracil-DNA glycosylase, UdgX, as a new biochemical tool to directly detect uracils in DNA
Authors: Jessica Stewart, Shanqiao Wei, Madhurima Datta, Umesh Varshney, Ashok Bhagwat
Journal: Cancer Research (2017)

Abstract The enzyme activation-induced deaminase (AID) is essential to the B lymphocyte development and antibody maturation. It converts cytosines in DNA to uracil. The unregulated expression of AID in many B cell cancers is correlated with an increased load of mutations. AID also promotes DNA strand breaks and is required for chromosomal translocations such as IgH/Myc in these tumors. We have previously showed that B cell cancer cell lines and patient tumors which express AID at high levels also have high levels of uracil in their genomic DNA. We accomplished this by excising the uracils using a uracil-DNA glycosylase (Ung) and labeled the resulting in an abasic site with an aldehyde reactive chemical (ARP). ARP contains a biotin tag and it was detected using a fluorescently labeled streptavidin. The results from this assay showed B cell tumors have as much as 100-fold higher levels of genomic uracils. Further development of this assay has allowed the detection of uracils in the blood plasma and buffy coat of B cell cancer patients. This “liquid biopsy” assay technique has the potential to work as a simple surveillance tool for B cell cancer patients who are in remission. In addition, we are exploring the possibility of a direct detection of uracil in DNA, which would provide greater sensitivity to this assay and simplify it. A unique uracil-DNA glycosylase was recently isolated from M. smegmatis, UdgX, that binds covalently to uracils in DNA. We are developing multiple ways to detect uracil-DNA-UdgX complexes and amplifying the resulting signal. We expect that this will allow us to directly visualize uracils in B-NHL tumors and detect uracils in circulating B-NHL tumor DNA from a drop of patient plasma. Citation Format: Jessica Stewart, Shanqiao Wei, Madhurima Datta, Umesh Varshney, Ashok Bhagwat. A novel uracil-DNA glycosylase, UdgX, as a new biochemical tool to directly detect uracils in DNA [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3802. doi:10.1158/1538-7445.AM2017-3802
Abstract 1480: Mir-182 is involved in sulindac anticancer activity in colon cancer
Authors: Hongyou Zhao, Bin Yi, Zhipin Liang, Ruixia Ma, Yaguang Xi
Journal: Cancer Research (2017)

Abstract Nonsteroidal anti-inflammatory drugs (NSAIDs) display promising antineoplastic activity in many human solid tumors including colorectal cancer. Previous studies reported that sulindac sulfide (SS) can inhibit the growth of tumor cells through cyclooxygenase-2 (COX-2) dependent or independent pathways. Obviously, COX-2 independent pathway involves a low toxic property to support the clinical potential for using sulindac as a chemoprevention drug. However, the molecular mechanisms responsible for COX-2 independent pathway have not been completely elucidated. In this project, we employed two human colon cancer cell lines, referred to HCT116 and HT29. HCT116 cells are characterized for low COX-2 expression, while HT29 cells show relatively high COX-2 expression. We found that SS could unbiasedly inhibit the growth of HCT116 and HT29 cells by arresting cells in G1/G2 phases. CyclinG2 was found to be upregulated in response to SS treatment. FOXO3a has been reported to regulate CyclinG2 expression at the transcriptional level. Our results demonstrate that SS could also upregulate FOXO3a. By using the loss-of-function strategy, we found that SS could not efficiently upregulate CyclinG2 and lead to cell cycle arrest in cells with FOXO3a knockdown. In addition, we studied the mechanism that could be involved in SS regulation of FOXO3a. Our previous studies reported that a panel of miRNAs could be altered by SS treatment in both colon and breast cancer cells. In the downregulated miRNA list, we found that miR-182 could potentially target FOXO3a. By using luciferase assay, we validated the direct regulation of miR-182 on the expression of FOXO3a. When miR-182 was downregulated, SS could neither efficiently upregulate the expression of FOXO3a nor inhibit cell growth as it did in control cells. We gained highly consistent results in HT29 and HT29 cells with COX-2 knockdown. Therefore, our study demonstrates a pathway consisting of miR-182/ FOXO3a/ CyclinG2 as a novel mechanism responsible for SS anticancer activity in colon cancer, which may imply a COX-2 independent pathway. Citation Format: Hongyou Zhao, Bin Yi, Zhipin Liang, Ruixia Ma, Yaguang Xi. Mir-182 is involved in sulindac anticancer activity in colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1480. doi:10.1158/1538-7445.AM2017-1480
Abstract 1383: Histone H3 lysine 9 methyltransferase SUV39H1 is associated with clinical outcome of renal clear cell carcinoma patients
Authors: Wei Meng, Ri Cui, Saikh Jaharul Haque, Carlo M. Croce, Arnab Chakravarti
Journal: Cancer Research (2017)

Abstract Background: Epigenetic alternations have three important components including DNA methylation, histione modifications, and non-coding RNA. The epigenetic changes are highly dynamic and also reversible, which regulate a variety of biological processes. Euchromatin which is a loosely packed form of chromatin is highly associated with active gene expression. In contrast, heterochromatin is associated with high histone methylation and DNA methylation but low RNA transcription. SUV39H1 catalyzes the histone 3 lysine 9 trimethylation (H3K9me3) in heterochromatin regions, which also mediates DNA methylation at pericentric heterochromatin. Currently, the oncogenic role of SUV39H1 in renal cell carcinoma (RCC) pathogenesis is poorly understood. To this end, we assessed the role of SUV39H1 in renal RCC cells and association with clinical outcome of RCC patients. Results: In SUV39H1 knocked down cells, we observed global H3K9me3 level reduction and cell growth inhibition. Proximal ligation assay was used to determine the H3K9me3 and 5-methylcytosine (5mC) interaction in RCC cells. Knocking down of SUV39H1 gene significantly reduced the co-localization of H3K9me3 and 5mC in pericentric repetitive sequences. Moreover, Southern blot demonstrated that expansion of a pentanucleotide (GGAAT) repeats was associated with upregulation of SUV39H1, which is linked to SUV39H1 function with microsatellite instability and rearrangement. In The Cancer Genome Atlas (TCGA) database, we found that SUV39H1 gene expression was upregulated in RCC tissues compared to normal tissues, and high SUV39H1 expression predicted a worse overall survival outcome (p = 0.002) in RCC patients. Conclusion: SUV39H1 upregulation promotes cell proliferation and microsatellite instability. SUV39H1 expression could serve as a prognostic biomarker for RCC patients. Citation Format: Wei Meng, Ri Cui, Saikh Jaharul Haque, Carlo M. Croce, Arnab Chakravarti. Histone H3 lysine 9 methyltransferase SUV39H1 is associated with clinical outcome of renal clear cell carcinoma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1383. doi:10.1158/1538-7445.AM2017-1383
Abstract LB-051: High LDHA expression predicts decreased survival in neuroblastoma
Authors: Carmen Dorneburg, Lisa Christner, Thomas F. Barth, Wolfgang Mueller-Klieser, Matthias Fischer, Hero Barbara, Anneleen Beckers, Frank Speleman, Klaus-Michael Debatin, Christian Beltinger
Journal: Cancer Research (2017)

Abstract Neuroblastoma (NB) is a common pediatric malignancy that often carries a poor prognosis. Aerobic glycolysis, i.e. the Warburg effect, is a hallmark of cancers, including NB. Lactate dehydrogenase A (LDHA) catalyzes the conversion of pyruvate to lactate. It is considered a pivotal component of the LDH tetramer, which is central in the Warburg effect. LDHA is overexpressed in many cancers. Little is known about the role of LDHA in NB. We therefore determined the expression of LDHA mRNA and protein in 651 and 112 clinically annotated patient samples, respectively. We show that enhanced mRNA expression and predominant cytoplasmic protein expression of LDHA, as detected by immunohistochemistry, is markedly associated with increased tumor aggressiveness and decreased patient survival. Surprisingly, despite the proposed pivotal role of LDHA in the Warburg effect, LDHA was dispensable for aerobic glycolysis in NB cells. Complete depletion of LDHA by CRISPR/Cas9-mediated knock-out in several NB cell lines did not decrease glucose consumption and production of pyruvate and ATP. In accordance, LDH activity was maintained despite loss of LDHA, as LDHB and LDHC were still expressed, most likely compensating for the loss of LDHA. In line with LDHA being dispensable for aerobic glycolysis in NB cells, LDHA depletion did not decrease malignant characteristics of NB cells. Thus, clonogenic growth, migration and tumorigenicity remained unaffected. Extending these findings from manifest to developing NB, we show that in the TH-MYCN NB mouse model the development of NB from normal sympathetic ganglia is not associated with increased LDHA expression. Taken together, this work establishes LDHA as a novel predictive marker in NB and calls into question the pivotal role in aerobic glycolysis attributed to LDHA. Citation Format: Carmen Dorneburg, Lisa Christner, Thomas F. Barth, Wolfgang Mueller-Klieser, Matthias Fischer, Hero Barbara, Anneleen Beckers, Frank Speleman, Klaus-Michael Debatin, Christian Beltinger. High LDHA expression predicts decreased survival in neuroblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr LB-051. doi:10.1158/1538-7445.AM2017-LB-051
Abstract 3229: Drug repurposing of topiramate in obesity-driven endometrial cancer
Authors: Arthur-Quan Tran, Stephanie A. Sullivan, Chunxiao Zhou, David Kaufman, Victoria Bae-Jump
Journal: Cancer Research (2017)

Abstract Introduction: Topiramate (TPM) is a B-D-fructopyranose sulfamate that acts by inhibiting carbonic anhydrases (CAs) and has been implicated as a novel inhibitor of angiogenesis. TPM is commonly used for the treatment of epilepsy and migraine headaches; however, when combined with phentermine, TPM can induce weight loss. Due to its weight loss and anti-angiogenic properties, we assessed TPM’s potential as an anti-tumorigenic agent in endometrial cancer (EC), a highly obesity-driven disease. Methods: Cell proliferation was assessed by MTT assay after exposure to TPM for 72 hours in the HEC-1A, KLE, Ishikawa and ECC-1 EC cell lines. Two representative cell lines, ECC-1 and Ishikawa, were used to analyze apoptosis, cell cycle progression, cell adhesion and invasion. Apoptosis was analyzed by Annexin V-FITC assay. Inhibition of adhesion and invasion by TPM were assessed by ELISA and transwell assay, respectively. Cell cycle progression was evaluated by Cellometer. Western immunoblotting was performed to assess downstream targets of the MAPK and mTOR pathways. The LKB1fl/flp53fl/fl EC mouse model was utilized to assess the in vivo effects of TPM. AdCre was injected at six weeks of age to induce invasive EC. Eight wks following AdCre injection, mice (N=10 per group) were treated with placebo or TPM (200 mg/kg/day, oral) for four weeks. The expression of phosphorylated-S6 and Ki-67 was assessed by IHC. Results: TPM inhibited cell proliferation in a dose dependent manner in all four EC cell lines (IC50 range=1500-3000 mM). Treatment with TPM resulted in G1 arrest and induction of apoptosis (p&amp;lt;0.05). TPM also reduced adhesion and invasion in the EC cell lines (p=0.05), with a corresponding decrease in VEGF expression. In addition, TPM significantly inhibited phosphorylation of p42/44 and S6 in the EC cell lines. Lastly, TPM decreased tumor weight in the LKB1fl/flp53fl/fl mice by 68% compared to those treated with placebo (p&amp;lt;0.05), accompanied by a reduction in Ki-67 and phosphorylated-S6 expression (p&amp;lt;0.05). Conclusion: TPM inhibited cell proliferation and tumor growth in EC cell lines and an EC mouse model. Therefore, TPM may be worthy of drug repurposing as an anti-tumorigenic agent in EC, with the potential added benefit of weight loss in this obesity-linked disease. Citation Format: Arthur-Quan Tran, Stephanie A. Sullivan, Chunxiao Zhou, David Kaufman, Victoria Bae-Jump. Drug repurposing of topiramate in obesity-driven endometrial cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3229. doi:10.1158/1538-7445.AM2017-3229
Abstract 66: Anthracycline-based antibody drug conjugates with potent immune-stimulatory functions
Authors: Roger R. Beerli
Journal: Cancer Research (2017)

Abstract Antibody drug conjugates (ADCs) are highly potent and selective anti-tumor drugs, combining the specific targeting of monoclonal antibodies with the potency of small molecule toxic payloads. Here, we employed enzymatic, site-specific conjugation to generate homogenous ADCs based on a derivative of the highly potent anthracycline toxin PNU-159682 and a non-cleavable peptide linker, using the anti-HER-2 antibody trastuzumab (part of trastuzumab emtansine) and the anti-CD30 antibody cAC10 (part of brentuximab vedotin). Characterization of the resulting ADCs in vitro and in vivo showed that they were highly stable and exhibited potencies exceeding those of ADCs based on conventional tubulin-targeting payloads, such as trastuzumab emtansine and brentuximab vedotin. Anti-tumor activity in an immune-competent host involved activation of the immune system, as shown by evaluation of a trastuzumab-PNU ADC in a Kadcyla-resistant HER2-positive orthotopic breast cancer model. Depletion of CD8 T cells severely reduced the anti-tumor activity of the ADC, demonstrating an important role for T cells in driving tumor regression. Furthermore, when tumor free animals were re-challenged with the same tumor, tumor growth was entirely inhibited in the absence of any further ADC administration, indicating the development of an immunological memory. In summary, we present a novel ADC format endowed not only with highly potent cytotoxicity, but also effective immune-stimulatory functions. Citation Format: Roger R. Beerli. Anthracycline-based antibody drug conjugates with potent immune-stimulatory functions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 66. doi:10.1158/1538-7445.AM2017-66
Abstract 4874: Application of quantitative analysis of mouse cell contamination and purification of human tumor cells in patient derived orthotopic xenograft mouse models of pediatric brain tumors
Authors: Frank K. Braun, Mari Kogiso, Lin Qi, Huiyuan Zhang, Yuchen Du, Holly B. Lindsay, Sibo Zhao, Sarah G. Injac, Xiao-Nan Li
Journal: Cancer Research (2017)

Abstract Background: Pediatric brain tumors continue to be among the most difficult to cure and are the leading cause of cancer-related deaths in children. Research investigating pediatric brain cancer is still impaired by the lack of cancer models. We established a large panel of orthotopic xenograft models as well as several paired permanent cell lines using in vivo and in vitro culture techniques. However an inherent risk of xenograft models is the contamination of isolates of human tumors with host (mouse) cells. Therefor strategies to correctly and timely identify the human/mouse ratio are essential. In this study we investigated Prostaglandin Receptor EP2 (PTGER2) as a useful target to identifying mouse cell contamination in PDOX models. Method: Species specific primers based on Alcoster et al., 2011 were used to detect PTGER2 in genomic DNA extracts of xenograft and cultured cells. Real-time qPCR was carried out using a StepOnePlus™ Real-Time PCR System. Reaction mix contained SYBR Green select master mix, respective primers and 5 ng of total genomic DNA in a total reaction volume of 10 μL. The qPCR conditions were as follows: 50°C-2 min, 95°C-10 min, 40 cycles of (95°C-15 sec, 61°C-1 min). Primer specificity was controlled in addition by analysis of amplified products on a 1.2% agarose gel. FACS was used to isolate human cells according to anti-Human HLA-ABC Ab and mouse-specific Ab cocktail surface staining and subjected to qPCR. Results: Various models of pediatric brain cancer (PXA, MB, GBM, EPN, ATRT, DIPG, PNET) established in our group were investigated for mouse cell contamination. Cells from xenograft isolates as well as from cultured xenografts were routinely analyzed. Our study confirms PTGER2 as a useful target for accurately estimate mouse cell contamination in brain cancer xenograft models. This further enabled us to improve detection and characterization of HLA-ABC/mouse-cocktail double negative population as seen in FACS. Conclusion: In conclusion we have successfully implemented an easy qRT-PCR approach to determine the human/mouse cell ration in any mixed cell population. Particular for PDOX models it is paramount to have reliable information on mouse cell contamination. Citation Format: Frank K. Braun, Mari Kogiso, Lin Qi, Huiyuan Zhang, Yuchen Du, Holly B. Lindsay, Sibo Zhao, Sarah G. Injac, Xiao-Nan Li. Application of quantitative analysis of mouse cell contamination and purification of human tumor cells in patient derived orthotopic xenograft mouse models of pediatric brain tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4874. doi:10.1158/1538-7445.AM2017-4874