bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1900: Hypoxia induces accumulation of lysosomal proteins within mitochondria.
Authors: Nakamura Yasuyuki , Yoshida Masaki , Kitamura Noriaki , Kamino Hiroki , Murai Ryuya , Saito Yuri , Sano Hitoya , Arakawa Hirofumi
Published: August 2014

Mitochondrial quality control (MQC) plays a critical role in maintaining a healthy status of mitochondria, preventing aging, cancer, and a variety of degenerative diseases. However, the mechanism largely remains unclear. Recently, we discovered a novel MQC mechanism regulated by Mieap, a p53-inducible protein, in which Mieap induces intramitochondrial lysosome-like organella that plays a critical role in the elimination of oxidized mitochondrial proteins (designated MALM for Mieap-induced accumulation of lysosome-like organelles within mitochondria) in response to mitochondrial damage (1-4). Here, we report a physiological role of the MALM function. Hypoxia dramatically induced accumulation of lysosomal proteins within mitochondria without the destruction of mitochondrial structure in both human normal mammary epithelial cells (HMEC4s) and mouse embryonic fibroblasts cells (MEFs). In response to the hypoxic stress, the MALM was inducible in ATG5-KO, ATG7-KO, and PINK1-KO MEFs, but not in p53-KO and Mieap-KO MEFs, suggesting that the phenomenon is completely different from canonical autophagy. In the MALM-deficient cells, hypoxia caused an increase of mitochondrial reactive oxygen species (ROS) generation and accumulation of oxidized proteins within the mitochondria. Surprisingly, the MALM-deficient cells in the hypoxic condition revealed enhanced activities of cell migration and invasion. This phenotype was totally cancelled by a ROS scavenger reagent, Ebselen. These results suggest that the MALM is a universal and fundamental function in the cell (at least from mouse to human), which plays a critical role in MQC under hypoxia. Alteration of the MALM function in cancer cells could facilitate cancer invasion and metastasis via mitochondrial ROS generated by unhealthy mitochondria.

1. Miyamoto Y et al. PLoS ONE 6: e16054, 2011

2. Kitamura N et al. PLoS ONE 6: e16060, 2011

3. Nakamura Y et al. PLoS ONE 7: e30767, 2012

4. Miyamoto T et al. Sci Rep 2: 379, 2012

Citation Format: Yasuyuki Nakamura, Masaki Yoshida, Noriaki Kitamura, Hiroki Kamino, Ryuya Murai, Yuri Saito, Hitoya Sano, Hirofumi Arakawa. Hypoxia induces accumulation of lysosomal proteins within mitochondria. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1900. doi:10.1158/1538-7445.AM2013-1900
Abstract 1913: FGFR1 amplification as a druggable target in lung squamous cell carcinoma.
Authors: Kwon Mi Jung , Lee Seung Eun , Han Joungho , Ahn Jin Seok , Park Keunchil , Kim Jhingook , Ahn Myung-Ju , Choi Yoon-La
Published: August 2014

Background: Focal gene amplification is a potential target for diagnosis and therapy in cancer. Fibroblast growth factor receptor-type 1 (FGFR1) may play a role in the pathogenesis of lung squamous cell carcinoma (LSCC) and amplification of the FGFR1 gene is considered a candidate target for treatment of LSCC. The aim of this study was to simultaneously explore copy number changes of FGFR1 and 23 other genes by multiplex ligation-dependent probe amplification (MLPA), to compare the results with fluorescent in situ hybridization (FISH) data, and determine the clinicopathological significance of gene amplification.

Methods: This study included 81 resected LSCC specimens obtained between 2008 and 2010, and 14 lung cancer cell lines. MLPA was performed using fresh tumor samples and cell lines and FISH was applied to formalin-fixed, paraffin-embedded tissues.

Results: MLPA showed amplification/gain of MYC (48.1% of LSCCs), AURKA (44.4%), MET (29.6%), KIT (21.0%), and FGFR1 (14.8%). FISH showed FGFR1 amplification in 22 LSCCs (27.2%) using a cut-off value of ≥5. There was high concordance between FISH and MLPA (85.2%). No association between FGFR1 amplification and any clinicopathological variables was observed; however, associations were identified between EGFR and FGFR1 FISH-amplified tumors; PDGFRA and poor differentiation; KDR, ABL1, and MET and age; and SMO and CCND2 and pT stages.

Conclusions: Our MLPA and FISH data showed that FGFR1 amplification is a common event, occurring in 15-27.2% of LSCCs. Other amplified genes may be also candidate targets for the treatment of LSCC.

Citation Format: Mi Jung Kwon, Seung Eun Lee, Joungho Han, Jin Seok Ahn, Keunchil Park, Jhingook Kim, Myung-Ju Ahn, Yoon-La Choi. FGFR1 amplification as a druggable target in lung squamous cell carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1913. doi:10.1158/1538-7445.AM2013-1913
Abstract 1935: Expression of human Piwi-like genes is associated with prognosis for soft tissue sarcoma patients and is different in gender.
Authors: Greither Thomas , Koser Franziska , Kappler Matthias , Bache Matthias , Lautenschläger Christine , Goebel Steffen , Holzhausen Hans-Jürgen , Wach Sven , Würl Peter , Taubert Helge W.
Published: August 2014

The family of Argonaute genes is essential for stem cell maintenance and RNA interference. Piwi-like genes as a subclass of the Argonaute genes are expressed mainly in the germline. However, these genes may be re-expressed in tumors, and expression of the Piwi-like genes is associated with prognosis in several types of tumors.

We analyzed the expression of

Piwi-like mRNAs (Piwi-like 2-4) in

125 soft tissue sarcoma (STS) samples by qPCRs. Statistical tests were applied to study the correlation of expression levels with tumor-specific survival for

STS patients.

In multivariate Cox's regression analyses, we detected that low Piwi-like

2 and Piwi-like 4 mRNA expression were significantly associated with a worse prognosis (RR = 1.87; p = 0.032 and

RR = 1.82; p = 0.039). In addition, low expression of both genes was associated with a 2.58-fold increased risk of tumor-related death (p = 0.01).

Next we analysed expression of Piwi-like genes gender specific. Piwi-like 4 and combined Piwi-like 2 and 4 mRNA levels correlated significantly with prognosis (RR = 3.53; p

= 0.002 and RR = 5.23; p = 0.004) only for female but not for male patients.

However, combined low Piwi-like 2 and

3 transcript levels were associated with worse survival (RR = 5.90; p = 0.02) for male patients.

In this study, we identified a significant association between the expression of Piwi-like 2 and 4 mRNAs and the tumor-specific survival of soft tissue sarcoma patients. A connection between gender and the impact of Piwi-like mRNA expressions on STS patients’ prognosis was shown for the first time.

Citation Format: Thomas Greither, Franziska Koser, Matthias Kappler, Matthias Bache, Christine Lautenschläger, Steffen Goebel, Hans-Jürgen Holzhausen, Sven Wach, Peter Würl, Helge W. Taubert. Expression of human Piwi-like genes is associated with prognosis for soft tissue sarcoma patients and is different in gender. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1935. doi:10.1158/1538-7445.AM2013-1935
Abstract 1961: Overexpression of miR-196b and HOXA10 characterize a poor-prognosis gastric cancer subtype.
Authors: Lim Jae Yun , Yoon Sun Och , Seol So-Young , Hong Soon Won , Kim Jong Won , Choi Seung Ho , Lee Ju-Seog , Cho Jae Yong
Published: August 2014

Background: Gastric cancer is composed of disease subgroups with heterogeneous clinical and biological behaviors. A previous study identified two subgroups with distinct gene-expression profiles strongly associated with prognosis. To identify molecular biologic differences between these two subgroups, we performed an integrative analysis of mRNA, microRNA, and protein expression.

Methods: Array technologies were used to generate microRNA and protein expression profiles of frozen tissue samples from 60 patients with gastric cancer. All patients underwent curative gastric resection in 2005 and distributed from stage I to IV and their clinical characteristics were collected. BRB-ArrayTools were used for microarray data analysis. Data was compared between gastric cancer tissue and normal gastric tissue and also between two different prognostic groups. Aberrantly expressed microRNA and associated mRNA in patients with poor-prognosis gastric cancer were validated by quantitative RT-PCR and tissue microarray, respectively.

Results: Four microRNAs were aberrantly expressed in gastric cancer tissue, especially poor prognostic group (P < 0.05). In the poor-prognosis subgroup, miR-196b showed the most significantly high expression patterns and also miR-135b, miR-93. On the contrary, miR-29c* was down-regulated. MiR-196b expression was positively correlated with HOXA10 expression (r = 0.726; P < 0.001), which was significantly increased in poor-prognosis patients (P < 0.001). HOXA10 protein-positive expression was identified in 56 of 368 gastric cancer tissue microarray samples. In reverse-phase protein array, 46/124 proteins were expressed differently (P < 0.05); COX2 (P < 0.001) and cyclin B1 (P = 0.017) were clearly over-expressed in the poor-prognosis group.

Conclusion: Integrative analysis of RNA and protein expression profile facilitates interpretation of the molecular biologic heterogeneity of cancer. Co-activation of miR-196b and HOXA10 characterized a poor-prognosis subgroup of patients with gastric cancer. As HOXA10 is involved in the proliferation of hematopoietic stem cells and progenitor cells, the relevance of hematopoietic progenitor cell and gastric cancer development/progression should be further investigated.

Citation Format: Jae Yun Lim, Sun Och Yoon, So-Young Seol, Soon Won Hong, Jong Won Kim, Seung Ho Choi, Ju-Seog Lee, Jae Yong Cho. Overexpression of miR-196b and HOXA10 characterize a poor-prognosis gastric cancer subtype. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1961. doi:10.1158/1538-7445.AM2013-1961
Abstract 1967: Rb dephosphorylation and cleavage in Apoptosis.
Authors: Antonucci Lisa , Lentine Brandon , De Leon Gabriel , Krucher Nancy A.
Published: August 2014

The Retinoblastoma protein (Rb) is important in the control of cell proliferation and apoptosis. Its activity is controlled by reversible phosphorylation on several serine and threonine residues. The role of Rb phosphorylation in the control of apoptosis is largely unknown, although several apoptotic stimuli result in dephosphorylation of Rb. We have recently shown that dephosphorylation of Threonine-821 of Rb is required for apoptosis triggered by UV radiation and by the cdk inhibitor, Roscovitine. Here we show that Roscovitine induced dephosphorylation of Threonine-821 occurred concomitant with C-terminal cleavage of Rb, which is a known marker of apoptosis. In attempt to characterize the role of Rb dephosphorylation in the process of Rb cleavage, we have utilized mouse embryonic fibroblasts (MEFs) that express a mutant Rb in which the carboxy-terminal caspase cleavage site (884DEADG888) of Rb is altered to DEAAE which is resistant to cleavage (RB-MI). Roscovitine treatment (8 hours, 25μM) causes a reduction in cell number in WT MEFs, but has no effect on RB-MI MEFs. In addition, Roscovitine causes apoptosis in 60-70% of WT MEFs, however Roscovitine is unable to induce apoptosis in RB-MI MEFs demonstrating that the cdk inhibitor requires the carboxy-terminal cleavage of Rb in order to stimulate apoptosis. Apoptosis was measured using a DNA fragmentation ELISA, TUNEL assay, and by immunoblotting of active caspase-3. Experiments are currently underway to determine whether dephosphorylation of Threonine-821, specifically, is required for Rb cleavage.

Citation Format: Lisa Antonucci, Brandon Lentine, Gabriel De Leon, Nancy A. Krucher. Rb dephosphorylation and cleavage in Apoptosis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1967. doi:10.1158/1538-7445.AM2013-1967
Abstract 1972: Clinical impact of 6q deletion patterns in prostate cancer.
Authors: Kluth Martina , Amschler Nina , Heinl Anna , Jung Simon , Simon Ronald , Schlomm Thorsten , Sauter Guido , Minner Sarah Jane Pauline
Published: August 2014

Deletions of chromosome 6q12-22 represent the second common deletion in prostate cancer. Although most of these deletions are large and span > 50 megabases, a minimal deleted region containing 3-4 megabases was described at 6q15. MAP3K7 (at 6q15) was suggested as a potential target gene in this area. Based on the hypothesis, that large 6q deletions compromising a multitude of genes may have a different impact on the biology of prostate cancer cells than small circumscribed 6q15 deletions, a tissue microarray (TMA) containing over 7000 prostate cancers was analyzed by fluorescence in situ hybridization (FISH). All cancers were analyzed with probes for 6q12, 6q14, 6q15 (MAP3K7), 6q16, and 6q22. 3888 tumors were interpretable for all 5 probes including 463 (10.6%) showing a 6q deletion. 6q deletions were tightly linked to a negative ERG status with 15.6% deletions in ERG negative and 5.8% deletions in ERG positive cancers (p<0.0001). The frequency of 6q deletions increased with Gleason grade and pT stage. 6q deletions were found in 5.4% of ≤3+3, 10.6% of 3+4, 19.6% of 4+3, and 18.8% of ≥4+4 cancers (p<0.0001). 6q deletions occurred in 9.6% of pT2, 11.5% of pT3a, and 14.8% of pT3b carcinomas (p=0.0160). 6q deletions were also linked to early PSA recurrence (p<0.0001). Remarkably, the clinical outcome varied markedly with deletion size. PSA recurrence was more likely in cancers with 6q12-22 deletions as compared to tumors with 6q14-16 deletions (p<0.0001). Twelve genes located within 6q14-16 were functionally evaluated applying colony formation assays to prostate cell lines BPH1, DU-145, and PC-3. These analyses failed to identify a cell behavior consistent with a role as a tumor suppressor gene for MAP3K7, as well as RRAGD, ANKRD6, LYRM2, MDN1, CASP8AP2, BACH2, COQ3, MANEA, RNGTT, SLC35A1, and MMS22L. In conclusion, the strong impact of the deletion size suggests, that multiple genes on chromosome 6q12-22 might have a tumor suppressive role in prostate cancer and that alterations of more than one of these has an additive adverse effect to cells. Our data do not confirm MAP3K7 as the main target of 6q deletions in prostate cancer

Citation Format: Martina Kluth, Nina Amschler, Anna Heinl, Simon Jung, Ronald Simon, Thorsten Schlomm, Guido Sauter, Sarah Jane Pauline Minner. Clinical impact of 6q deletion patterns in prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1972. doi:10.1158/1538-7445.AM2013-1972
Abstract 1974: Functional characterization of Basonuclin 1 (BNC1): a novel tumor suppressor gene commonly downregulated in human prostate cancer.
Authors: Kwabi-Addo Bernard , Wang Songping , Furbert-Harris Paulett , Yegnasubramanian Srinivasan , Devaney Joseph
Published: August 2014

Prostate cancer (PCa) harbors a myriad of aberrant genomic and epigenetic alterations. Epigenetic inactivation of genes in PCa is largely based on transcriptional silencing by aberrant CpG methylation of CpG rich promoter regions. To better understand the role of aberrant methylation changes and to identify biological pathways likely to be affected in PCa, we used a genome-wide methylation array from Illumina (Infunium) to identify novel methylated genes associated to PCa. We identified several candidate genes including BNC1 as novel genes which are frequently methylated and whose methylation closely related to inactivation of gene expression in prostate cancer cell lines. Basonuclin1 is a zinc finger transcription factor that interacts with a subset of promoters of genes transcribed by both RNA polymerase-I and -II and has roles in maintaining ribosomal biogenesis and the proliferative potential of immature epithelial cells. We found that the expression of BNC1 negatively correlated with the degree of methylation at the CpG island in prostate tumor samples compared to benign prostate tissues. The forced-expression of BNC1 significantly decreased prostate cancer cell proliferation whereas the siRNA mediated BNC1 knock-down increased prostate cancer cell proliferation. Overall, our data suggest a potential tumor suppressor role for BNC1 that includes regulation of cell proliferation and cell-cell interaction in prostate carcinogenesis.

Citation Format: Bernard Kwabi-Addo, Songping Wang, Paulett Furbert-Harris, Srinivasan Yegnasubramanian, Joseph Devaney. Functional characterization of Basonuclin 1 (BNC1): a novel tumor suppressor gene commonly downregulated in human prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1974. doi:10.1158/1538-7445.AM2013-1974
Abstract 1979: Tripartite motif 14 (TRIM14) is a putative tumor suppressor gene in non-small-cell lung cancer.
Authors: Hai Josephine , Tsao Ming-Sound
Published: August 2014

Background: Non-small-cell lung carcinoma (NSCLC) accounts for 85% of all malignant lung tumors. Our group previously identified Tripartite Motif 14 (TRIM14) as a component of a prognostic multigene expression signature for NSCLC patients. TRIM14 belongs to a conserved family of Tripartite Motif-encoding genes that are involved in a broad range of biological processes, such as transcriptional regulation, cell proliferation, and apoptosis. However, TRIM14 itself is poorly understood, especially in the context of cancer progression. Here we investigate the biological function of TRIM14 in NSCLC.

Methods: The expression of TRIM14 was modified in human NSCLC cell lines (NCI-H1395, NCI-H1650 and NCI-H157) by lentivirus-mediated overexpression and short-hairpin RNA-mediated silencing. Effects were assessed by examining in vitro cell proliferation and anchorage-independent growth, and in vivo tumorigenicity in mice.

Results: In vitro studies show that TRIM14 overexpression in NSCLC cells suppresses proliferation and anchorage-independent growth, whereas knock-down of TRIM14 expression promotes cell proliferation and colony formation. In vivo studies demonstrate that knock-down of TRIM14 expression significantly increases tumor growth in immunodeficient mice.

Conclusion: This is the first study to suggest that TRIM14 may function as a tumor suppressor gene in lung cancer, affecting cell proliferation and anchorage-independent growth in vitro and tumor growth in mice.

(Supported by Canadian Cancer Society Research Institute grant #020527)

Citation Format: Josephine Hai, Ming-Sound Tsao. Tripartite motif 14 (TRIM14) is a putative tumor suppressor gene in non-small-cell lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1979. doi:10.1158/1538-7445.AM2013-1979
Abstract 198: Resveratrol's Metabolomic biomarkers of efficacy: the missing link to chemoprevention.
Authors: Saud Shakir , Young Matthew , Karoly Ed , Bernard Chris , Colburn Nancy , Kim Young
Published: August 2014

Chemoprevention has received considerable attention over the last decade. The notion of using bioactive food components to reduce the risk of developing, or the recurrence of cancer has long been recognized. Resveratrol (a naturally occurring polyphenolic compound) has been studied extensively with considerable evidence demonstrating its efficacy as an anti-inflammatory and anti-cancer agent, with the most evidence for its role in colon cancer prevention. However, Resveratrol intervention studies in clinical trials still remains controversial, due to the lack of biomarkers of efficacy. Utilizing Metabolomic analysis we have identified biomarkers of efficacy in a pre-clinical model of inflammation-induced colorectal carcinogenesis.

Metabolomics is the process of identifying metabolites present in biological specimens. Our study has demonstrated the chemopreventative activity of Resveratrol in mice treated with the chemical carcinogen azoxymethane (AOM) and then exposed to colonic irritant dextran sodium sulfate (DSS) to induce inflammation-associated colon carcinogenesis. We found a diet enriched in Resveratrol resulted in a reduction in tumor incidence in AOM/DSS treated mice by 29 percent. Furthermore, we identified several biomarkers that are indicative of Resveratrol chemopreventive activities. Prior studies have shown Resveratrol's role in inhibiting several key enzymes important during the carcinogenesis process. We have correlated such findings with Metabolomic biomarkers. We found a significant reduction in Arachidonic acid, the limiting reagent of pro-inflammatory enzyme COX-2, and a reduction in spermidine a product of Ornithine decarboxylase, an enzyme that promotes proliferation. More intriguingly, we found biomarkers representing changes in bile acids by Resveratrol, which highlight chemopreventative activities that were not previously described. We found that Resveratrol treatment causes a reduction in tumorigenic secondary bile acids, deoxycholic acid and lithocholic acid, which are metabolic byproducts of intestinal bacteria. These data suggested that dietary supplementation with Resveratrol results in a healthy microbiome.

These pre-clinical studies should easily be translated into clinical studies designed to ascertain chemopreventive efficacy for bioactive compounds like Resveratrol, all with the objective to identify patients that will benefit from an intervention.

Citation Format: Shakir Saud, Matthew Young, Ed Karoly, Chris Bernard, Nancy Colburn, Young Kim. Resveratrol's Metabolomic biomarkers of efficacy: the missing link to chemoprevention. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 198. doi:10.1158/1538-7445.AM2013-198
Abstract 1984: The Ikaros tumor suppressor regulates transcription of genes involved in cell signaling in human leukemia.
Authors: Payne Jonathon L. , Sloane Justin , Bamert Rebekka M. , Dovat Elanora , Li Hui , Bulathsinghala Marie , Gowda Chandrika , Song Chunhua , Dovat Sinisa
Published: August 2014

The IKZF1 gene encodes Ikaros, a DNA binding zinc finger protein that is essential for normal hematopoietic differentiation. Deletion or mutation of Ikaros has been associated with a both T and B-cell acute lymphoblastic leukemia (ALL), as well as acute and chronic myelogenous leukemias (AML and CML). Deletion of an Ikaros allele has also been identified as a poor prognostic marker in childhood B-ALL. Thus, Ikaros is established as a major tumor suppressor in hematopoietic malignancies. The Ikaros protein can activate or repress target gene expression via chromatin remodeling. The specific mechanisms by which Ikaros exerts its tumor suppressor activities are unknown. We have defined genome-wide occupancy of Ikaros in human leukemia using ChIP-SEQ and identified a large number of potential Ikaros target genes. The quantitative Chromatin Immunoprecipitation assay (qChIP) was used to identify genes whose expression is directly regulated by Ikaros. Using this technique we show that Ikaros binds to the promoter region of a gene that is part of a pro-oncogenic pathway, as well as a tumor suppressor gene. Both genes are essential components of cell signaling pathways. The promoter regions of the two novel target genes were cloned into luciferase reporter constructs and used to transiently transfect HEK 293T cells that were co-transfected with Ikaros or empty vector (as a control). Luciferase reporter assays demonstrate that Ikaros suppresses transcription of the gene that is involved in oncogenic transformation, while it positively regulates transcription of the tumor suppressor gene. In summary, we have identified two novel Ikaros target genes. Our results suggest that Ikaros exerts its tumor suppression function by regulating the expression of target genes involved in pro-oncogenic and tumor suppressor signaling in leukemia cells. These data identify mechanisms of the tumor suppressor activity of Ikaros in leukemia.

Citation Format: Jonathon L. Payne, Justin Sloane, Rebekka M. Bamert, Elanora Dovat, Hui Li, Marie Bulathsinghala, Chandrika Gowda, Chunhua Song, Sinisa Dovat. The Ikaros tumor suppressor regulates transcription of genes involved in cell signaling in human leukemia. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1984. doi:10.1158/1538-7445.AM2013-1984
Abstract 1987: Tumor suppressor ADAM23 mediates the HPV-associated carcinogenesis.
Authors: Yang Paul , Williams Drake W. , Kim Terresa , Shin Ki-Hyuk , Kang Mo , Park No-Hee , Kim Reuben H.
Published: August 2014

“High risk” human papillomaviruses (HPV) are known to cause or are closely associated with the development of human cervical, anal and some types of head and neck cancers. Integration of HPV DNA into host genome and subsequent expression of viral oncogenes, such as E6 and E7, are critical initial events for the HPV-associated cancer development; however, HPV infection alone is not sufficient to convert normal cells to malignant phenotypes, suggesting that other additional genetic and/or epigenetic events in the HPV-infected cells are required. The aim of the current study is to identify molecular determinants that are functionally linked to the pathogenesis of the HPV-associated cancers. We used HPV-16 harboring immortalized but non-transformed oral epithelial cells (HOK-16B), infected with high-throughput Decode RNAi lentiviral screening pools containing about 70,000 shRNAmir constructs capable of knocking down the target genes individually (3-4 constructs per gene), and selected them in DMEM-based high calcium/serum media in which HOK-16B cells are not permissive to survive. From the surviving colonies, we have identified several putative tumor suppressors including A Disintegrin and etalloproteinase domain-containing protein 23 (ADAM23). The function of ADAM23 was further validated such that singly knocking down in HOK-16B cells (HOK16B/ADAM23i) acquired proliferative potential in DMEM-based medium. HOK16B/ADAM23i cells also gained resistance to ionizing radiation (IR), cisplatin treatment, and differentiation as demonstrated by colonogenic assay, MTT assay, and the 3-dimensional raft culture systems, respectively. More importantly, the expression is largely diminished in squamous cell carcinoma (SCC) cell lines but more so in HPV-harboring SCC. We further found that ADAM23 expression is high in normal human oral epithelial tissues but low or none in HPV-positive OSCC tissue specimens as determined by immunohistochemical staining. Our data suggest that ADAM23 may play a role in development of HPV-associated cancers.

Citation Format: Paul Yang, Drake W. Williams, Terresa Kim, Ki-Hyuk Shin, Mo Kang, No-Hee Park, Reuben H. Kim. Tumor suppressor ADAM23 mediates the HPV-associated carcinogenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1987. doi:10.1158/1538-7445.AM2013-1987
Abstract 1993: Identification of genetic interactors of BRCA2 in mouse embryonic stem cell model.
Authors: Ding Xia , Sharan Shyam
Published: August 2014

Breast cancer is the most prevalent cancer in women worldwide and mutations in breast cancer susceptible genes, BRCA1 and BRCA2 are the genetic factors conferring highest risk (40-70%) of developing breast cancer. Although BRCA proteins are known to maintain genomic stability mainly by homologous recombination-mediated DNA repair, detailed mechanisms of how BRCA loss induces tumorigenesis remain unclear. Interestingly, while inactivation of both alleles of BRCA1 or BRCA2 genes is necessary for tumor development, their loss in embryonic cells affects cell viability. To explain this paradox, we hypothesize that cells lacking BRCA1 or 2 are able to survive and are predisposed to tumorigenesis due to mutations in other genes such as those involved in cell cycle regulation or DNA damage response. In the current study, we propose to identify such genes using a mouse embryonic stem (mES) cell-based insertional mutagenesis screen. In mES cells with one mutant allele of Brca2 and the other flanked by loxP sites, cell death can be induced by Cre-mediated loss of the conditional allele. To identify genes whose up-regulation can rescue BRCA2 loss-induced cell death, we transduced these mES cells with murine stem cell retrovirus (MSCV) expressing Cre recombinase. We obtained Brca2-null ES cells that were rescued by the viral insertion and used them to clone the viral insertional sites. We identified several candidate genes that were upregulated in viable ES cells. One of these genes encodes a PDZ domain-containing protein, GIPC3. Expression of Gipc3 was elevated 3 to 4-fold in the rescued clone. Furthermore, we rescued the lethality of Brca2-null ES cells by overexpressing Gipc3. The rescued cells were hypersensitive to DNA damaging agents and exhibited an overall increase in genomic instability. Mechanistically, we found DNA damage-induced p21 expression to be attenuated in GIPC3 over-expressing clones. Because p21 is a key mediator of BRCA2 loss-induced cell death, we hypothesize that GIPC3 overexpression may inhibit p21 expression resulting in viable Brca2-null ES cells. We are now studying the detailed signaling pathway of how GIPC3 affects p21 expression. In addition, we are examining if Gipc3 overxpression can rescue the lethality of Brca2ko/ko embryos and contribute to tumorigenesis in Brca2ko/+ mice. We are also examining the expression of GIPC3 in human BRCA2-deficient tumors.

Citation Format: Xia Ding, Shyam Sharan. Identification of genetic interactors of BRCA2 in mouse embryonic stem cell model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1993. doi:10.1158/1538-7445.AM2013-1993
Abstract 2010: Whole genome profiling to identify novel metastasis regulators in colorectal cancer.
Authors: Hauser Christine , Shavinskaya Anna , Hutter Barbara , Abba Mohammed , Tkachenko Olga , Lichter Peter , Eils Roland , Brors Benedikt , Allgayer Heike
Published: August 2014

Colorectal cancer is the second most common cancer and the second leading cause of cancer related death in men and women in Germany according to the Robert Koch Institute. The leading cause for cancer related mortality is the development of metastasis.

In our study, we aim to identify novel regulators of the metastatic process in colorectal cancer. To this end we employed next generation whole genome sequencing of resected tissues from patients with metastatic colorectal cancer. In particular, we sequenced samples from primary tumor, corresponding normal colon mucosa, metastasis and normal tissue from the metastatic site. The DNA from one patient was thereby sequenced on two different platforms, one being Illumina HiSeq the other Complete Genomics (CG). Additionally, we sequenced the transcriptome of this patient to correlate mRNA expression with genetic aberrations. This abstract focusses on the first results regarding metastatic aberrations.

The average read coverage for the samples sequenced with the Illumina platform was between 50-60x and for the samples sequenced with CG between 105-135x. In total 3607 SNVs were identified in coding regions in the metastasis with both platforms, 40 of these were in exonic regions and classified as high confidence with both platforms.

We also sequenced samples from an additional two patients on one sequencing platform each. One of the patients also presented with liver metastasis, the other one with a lung metastasis. These data were also compared to the data from the first patient to identify possible common aberrations as well as possible differences between the different metastatic sites. We identified several copy number changes occurring in the same chromosomal regions in different patients. Similarly, several genes affected by SNVs were common in two of the patients, while only high confidence APC mutations were common to all three patients. Seven additional genes were affected by high confidence SNVs in the two patients sequenced using the CG platform and three on the Illumina platform, two of which have not yet been characterized in the context of cancer or metastasis.

Citation Format: Christine Hauser, Anna Shavinskaya, Barbara Hutter, Mohammed Abba, Olga Tkachenko, Peter Lichter, Roland Eils, Benedikt Brors, Heike Allgayer. Whole genome profiling to identify novel metastasis regulators in colorectal cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2010. doi:10.1158/1538-7445.AM2013-2010
Abstract 2024: Identification of novel oncogenes in a stage I small cell lung cancer through whole-genome sequencing.
Authors: Han Ji-Youn , Lee Yeon Su , Kim Byung Chul , Lee Sung Hoon , Lee Geon Kook , Cho Youn-Sung , Kim Hak-Min , Lee Jin Soo
Published: August 2014

Background: Small cell lung cancer (SCLC) is rapidly progressive and most cases are presented with advanced stage. It hampers access to early stage cancer samples, which are essential to understand the early development of this disease. We performed whole-genome sequencing (WGS) of a stage I SCLC to analyze the genomic feature of early tumorigenesis of SCLC.

Methods: We obtained normal and tumor genomic DNAs from a patient with stage I SCLC underwent curative resection and analyzed the normal and cancer genomes by WGS using 90 bp paired-end reads and validated the cancer variations by Sanger sequencing and single-nucleotide polymorphism (SNP) genotyping.

Results: WGS revealed a lot of SNVs, small insertion/deletions, and chromosomal abnormality. Chromosomes 4p, 5q, 13q, 15q, 17p, and 22q contained many block deletions. Especially, copy loss was observed in tumor suppressor genes RB1 and TP53, and copy gain in oncogene hTERT. Recurrent somatic mutations were found in two genes, TP53 and CREBBP. Novel non-synonymous (ns) single nucleotide variations (SNV) were found in C6ORF103 and SLC5A4 genes. Sequencing of SLC5A4 gene in 23 independent limited disease SCLC samples revealed another ns SNV in SLC5A4 gene, indicating ns SNVs in SLC5A4 gene are recurrent in early -stage SCLC. This cancer genome had mutated genes associated with Notch and WNT signaling pathways.

Conclusion: WGS analysis in this study provides the genomic landscape of early-stage SCLC, which may provide new insight into the early development of SCLC.

Citation Format: Ji-Youn Han, Yeon Su Lee, Byung Chul Kim, Sung Hoon Lee, Geon Kook Lee, Youn-Sung Cho, Hak-Min Kim, Jin Soo Lee. Identification of novel oncogenes in a stage I small cell lung cancer through whole-genome sequencing. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2024. doi:10.1158/1538-7445.AM2013-2024
Abstract 2029: Interactions between a selenophene compound and DNA polymerase eta.
Authors: Rietz Anne , Chen Yih-wen , Chou Kai-Ming
Published: August 2014

D-501036 [2,5-bis(5-hydroxymethyl-2-selenienyl)-3-hydroxymethyl-N-methylpyrrole] is a selenophene compound that has potent toxicity to various types of cancer cells but not for normal cells. More importantly, in vivo xenograft animal studies have also shown that D-501036 is a promising anticancer compound. Mechanistic studies indicated that D-501036 induces DNA double strand breaks in a dose dependent manner and blocks cell cycle progression at S-phase. D-501036 forms DNA adducts and the amount of DNA adducts increased with the concentration of D-501036. Therefore, we examined several DNA replication/repair mechanisms for their potential relationship with D-501036. Interestingly, we observed that D-501036 suppressed the expression of DNA lesion bypass polymerase η (pol η) more than 70% in both time and dose dependent manners in the ovarian Ovcar-4 cancer cells.

Pol η is the key enzyme to replicate across UV induced cyclobutane pyrimidine dimers (CPD) in DNA. Genetic defects in the pol η encoding gene result in a disease called Xeroderma Pigmentosum variant (XP-V). In addition to its biological function, biochemical studies have shown that pol η can also efficiently bypass cisplatin intrastrand crosslinked guanosines (Pt-GG). In consistence with the biochemical studies, cellular studies have shown that the expression of pol η reduces the cytotoxicity of cisplatin in Ovcar-4 cells. These results suggested that pol η plays an important role in modulating the cellular sensitivity to cisplatin. Although cisplatin is an important therapeutic agent for the treatment of ovarian cancer but the development of resistance against cisplatin has been a major drawback.

Given that D-501036 suppresses the expression of pol η, we performed a cytotoxicity study using the combination of D-501036 and cisplatin. Our results indicated that D-501036 can potentiate the sensitivity of Ovcar-4 cells to cisplatin more than 1,000- fold. Together, our results suggested that D-501036 is a promising compound that can be used to enhance the activity of cisplatin for the treatment of ovarian cancer. Further studies are undergoing to determine the mechanisms of action of D-501036, the information obtained will facilitate the development of new compounds to improve the treatment for ovarian cancer.

Citation Format: Anne Rietz, Yih-wen Chen, Kai-Ming Chou. Interactions between a selenophene compound and DNA polymerase eta. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2029. doi:10.1158/1538-7445.AM2013-2029
Abstract 2056: Anticancer bioactive peptide potentiates cisplatin chemotherapy efficacy to improve the quality of life in xenografted nude mice bearing human gastric cancer.
Authors: Su Xiulan , Dong Chao , Su Liya , Zhang Jialing , Wang Xuemei , Bai Xia , Cui Hongwei , Chen Zhong
Published: August 2014

A great challenge of cancer chemotherapy is to eliminate cancer cells and concurrently maintain the quality of life (QOL) for cancer patients. We previously found that anticancer bioactive peptide (ACBP) alone exhibited anti-tumor activity without measurable side effects. Thus, we hypothesize that ACBP and combined chemotherapy could lower the drug dose and improve the efficacy of treatment, leading to a better QOL. In this study, ACBP was isolated and purified from goat liver, and designated as ACBP-L. The anti-tumor activity of ACBP-L was titrated on human gastric cancer MGC-803 cell line in vitro by MTT assay, and its effect on cell morphology was observed through light and scanning electron microscopy. In vivo, MGC-803 tumors were grown in a xenograft nude mouse model, the body weight, food intake, volume and weight of tumors were measured. 18F- FDG PET/CT scan was used to examine the biological activity in tumors of live animals. Expression of Bcl-2, Bax, Caspase 3 and Caspase 8 in the tumor specimens was examined by RT-PCR and immunohistochemistry. Herein, we showed that ACBP-L inhibited MGC-803 cells in a dose and time dependent manner in vitro. In the MGC-803 xenograft nude mouse model, ACBP-L alone significantly inhibited tumor growth in vivo without measurable side effects. Treatment with the full dosage of Cisplatin alone (5mg/kg every 4 days) strongly suppressed tumor growth; however, the QOL in these mice had been significantly affected when measured by food intakes and body weight. The combinatory regiment of ACBP-L with a fewer doses of Cisplatin (5mg/kg every 10 days) resulted in a similar anti-tumor activity with improved QOL. The tumor tissues were harvested after treatment, and ACBP-L suppressed Bcl-2, and induced Bax, Caspase 3, and Caspase 8 protein expression more significantly than in the tissues harvested from Cisplatin alone treatment as detected by immunohistochemistry. The combinatory regiment induced stronger gene expression of Bax and Caspase 8. Our data demonstrated that ACBP-L could lower Cisplatin dose to achieve a similar anti-tumor efficacy as the higher dose of Cisplatin alone. This combined regiment improved QOL in the xenograft gastric cancer model through the modulation of apoptotic molecules.

Citation Format: Xiulan Su, Chao Dong, Liya Su, Jialing Zhang, Xuemei Wang, Xia Bai, Hongwei Cui, Zhong Chen. Anticancer bioactive peptide potentiates cisplatin chemotherapy efficacy to improve the quality of life in xenografted nude mice bearing human gastric cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2056. doi:10.1158/1538-7445.AM2013-2056
Abstract 2067: BMN 673 potentiates efficacy of temozolomide in sensitive and resistant glioblastoma models.
Authors: Kizilbash Sani H. , Kawashima Ryo , Gupta Shiv , Mladek Ann C. , Shen Yuqiao , Sarkaria Jann N.
Published: August 2014

Poly (ADP-ribose) polymerase (PARP) plays an important role in several DNA repair processes, and previous studies with various PARP inhibitors have demonstrated robust enhancement of sensitivity to temozolomide (TMZ) chemotherapy in pre-clinical models of glioblastoma multiforme (GBM). The focus of this study was to evaluate the antitumor effects of a novel PARP inhibitor, BMN 673, as a single agent and in combination with TMZ in the established U251 glioma cell line, neurosphere cultures from the primary Mayo GBM xenograft line GBM12, and TMZ-resistant cells derived from either line (U251TMZ, GBM12TMZ(MGMT high) and GBM12TMZ(MGMT low)). In a biochemical assay of PARP inhibition, BMN 673 potently suppressed poly-ADP-ribosylation of cellular proteins at concentrations of 3 nM following hydrogen peroxide treatment of U251 cells. Single agent treatment had modest anti-tumor activity with 30 nM BMN 673 reducing absorbance in a CyQUANT cell proliferation assay relative to control treatment in both U251 and U251TMZ cells (Table 1). BMN 673 co-treatment also enhanced the efficacy of TMZ in both U251-derivative lines at a clinically relevant concentration of TMZ. Similarly, treatment with BMN673 alone or in combination with TMZ inhibited primary neurosphere formation in all three GBM12 models. Notably, the addition BMN 673 clearly re-sensitized both resistant GBM12 sub-lines to relatively low concentrations of TMZ. Consistent with a mechanism involving disruption of DNA repair, combined treatment with BMN 673 and TMZ resulted in robust activation of ATM-mediated KAP1 phosphorylation (Ser824) and ATR-mediated Chk1 phosphorylation (Ser345) as compared to TMZ or BMN 673 alone 24 hours after treatment. In conclusion, BMN673 is a promising PARP inhibitor with high potency and cytotoxicity in vitro against both TMZ sensitive and TMZ resistant GBM cells.

Table 1 - In vitro antitumor effects of TMZ combined with BMN673 TMZ, microM 0 nM BMN673 30 nM BMN673 CyQUANT U251 Relative absorbance Relative absorbance 0 1.00 0.58 30 0.80 0.30 U251TMZ 0 1.00 0.69 30 1.02 0.21 Primary Neurosphere Assay GBM12 Relative Count Relative Count 0 1.00 0.66 5 0.32 0 10 0.01 0 GBM12TMZ (MGMT high) 0 1.00 0.32 5 1.11 0.02 10 1.04 0.01 GBM12TMZ (MGMT low) 0 1.00 0.25 5 0.86 0.03 10 0.75 0

Citation Format: Sani H. Kizilbash, Ryo Kawashima, Shiv Gupta, Ann C. Mladek, Yuqiao Shen, Jann N. Sarkaria. BMN 673 potentiates efficacy of temozolomide in sensitive and resistant glioblastoma models. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2067. doi:10.1158/1538-7445.AM2013-2067
Abstract 2097: Targeting aneuploidy with NMS-P153, a tight binder inhibitor of the spindle assembly checkpoint MPS1 (TTK) kinase.
Authors: Colombo Riccardo , Caldarelli Marina , Giorgini Maria Laura , Degrassi Anna , Ciomei Marina , Pezzetta Daniele , Ballinari Dario , Montagnoli Alessia , Pesenti Enrico , Donati Daniele , Galvani Arturo
Published: August 2014

Aneuploidy is one of the major hallmarks of cancer and is a unique property of tumoral cells. Targeting of the molecular mechanisms required for growth of aneuploid cells is a potential strategy for developing new therapies with preferential activity against cancer.

The Spindle Assembly Checkpoint (SAC) is a mitotic mechanism required for proper chromosomal segregation, ensuring that cells do not divide until all sister chromatids correctly align to the metaphase plate. The SAC is a complex machinery regulated by a number of members which are rarely mutated in tumors. Rather, checkpoint components have been found to be upregulated in aneuploid tumor cells, suggesting a potential tumor “addiction” to SAC functions.

MPS1 kinase, (also known as TTK) is an essential enzyme of the SAC complex which is aberrantly overexpressed in a wide range of tumors and necessary for tumor cell proliferation. We previously reported the identification of NMS-P715, a selective and orally bioavailable MPS1 inhibitor, capable of promoting mitotic acceleration, thereby causing massive aneuploidy and cancer cell death both in vitro and in vivo and which promotes tumor growth inhibition in preclinical cancer models. Supported by these results, a lead optimization program was then undertaken which led to the identification of NMS-P153, an optimized product candidate for clinical development.

NMS-P153 is selective, shows high in vitro enzymatic potency on MPS1 (IC50 of 3 nM) and a prolonged residence binding time.

More than 100 cell lines have been analyzed against NMS-P153 in proliferation studies and several distinct xenograft models have been tested in vivo, allowing the identification of potential genetic contexts of sensitivity. Potent anti-proliferative activity, good tumor growth inhibition associated with tumor regression and biomarker modulation was observed in sensitive xenograft models.

A favorable ADME profile, associated with permissive therapeutic safety margins observed in different species at efficacious exposures, indicate that NMS-P153 is suitable for further development, with the potential for being a first-in-class drug targeting MPS1 kinase.

Citation Format: Riccardo Colombo, Marina Caldarelli, Maria Laura Giorgini, Anna Degrassi, Marina Ciomei, Daniele Pezzetta, Dario Ballinari, Alessia Montagnoli, Enrico Pesenti, Daniele Donati, Arturo Galvani. Targeting aneuploidy with NMS-P153, a tight binder inhibitor of the spindle assembly checkpoint MPS1 (TTK) kinase. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2097. doi:10.1158/1538-7445.AM2013-2097