bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2918: Peptide probe-based in vivo imaging of apoptosis and early decision on stomach tumor response to treatment
Authors: Khan Fatima , Gurung Smriti , Jung Hyun-Kyung , Baek Moon-Chang , Choi Yong Woo , Kim In San , Lee Byung Heon
Published: August 2015

Early decision on tumor response after anti-cancer treatment is still an unmet medical need. Here we exploited ApoPep-1 peptide that binds to histone H1 exposed on apoptotic cell surface as a probe and investigated whether in vivo imaging of apoptosis using near-infrared fluorescence dye-labeled linear and cyclic (disulfide-bonded) form of ApoPep-1 at an early stage after treatment could enable us predict tumor response to the treatment later. Treatment of SNU16 stomach tumor cells with cistplatin and cetuximab alone significantly induced apoptosis of the cells, while combination of cisplatin and cetuximab more efficiently induced apoptosis, as detected by staining with linear and cyclic form of ApoPep1. However, the differences among groups were more remarkable when detected with cyclic form of ApoPep-1 compared to the linear counterpart. In tumor-bearing mice, apoptosis imaging was performed at the first and second week after the initiation of treatment, and tumor sizes and weights were measured 3 weeks after treatment. In vivo fluorescence imaging signals by the homing of ApoPep-1 to tumor was most remarkable in the group injected with cyclic form of ApoPep-1 at the first week after combined treatment with cisplatin plus cetuximab. There were no significant differences in tumor sizes and weights between groups injected with linear and cyclic form of ApoPep-1. Correlation analysis revealed that imaging signals 1 week after treatment with cisplatin and cetuximab in combination were most closely related with tumor volume changes (r2 = 0.934). These results demonstrate that in vivo apoptosis imaging using the cyclic form of Apopep-1 is a sensitive and predictive tool for early decision on stomach tumor response after anti-cancer treatment.

Citation Format: Fatima Khan, Smriti Gurung, Hyun-Kyung Jung, Moon-Chang Baek, Yong Woo Choi, In San Kim, Byung Heon Lee. Peptide probe-based in vivo imaging of apoptosis and early decision on stomach tumor response to treatment. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2918. doi:10.1158/1538-7445.AM2015-2918
Abstract 2925: WAVE3 is associated with chemoresistance in TNBC by activating tumor angiogenesis downstream of the STAT1/Hif-1α/VEGF-A signaling axis
Authors: Davuluri Gangarao , Plow Edward F. , Schiemann William P. , Sossey-Alaoui Khalid
Published: August 2015

Chemoresistance is the main cause of disease recurrence and eventual death of breast cancer patients, particularly those harboring triple-negative breast cancer (TNBC) tumors for which effective FDA-approved drugs currently do not exist. However, a recent study determined that TNBCs can be segregated into 6 genetically distinct subtypes that do in fact exhibit differential rates of pathological complete response (pCR) to standard-of-care chemotherapies. Interestingly, the mesenchymal and mesenchymal stem-like subtypes of TNBCs exhibit the lowest rates of pCR when treated with standard-of-care chemotherapies. WAVE3 is an actin-cytoskeleton remodeling protein, and recent studies have highlighted a potential role for WAVE3 in promoting tumor progression and metastasis in TNBC. However, whether WAVE3 activity is involved in the development of chemoresistance in TNBCs, remains unclear. Here we show that loss of WAVE3 expression resensitizes human TNBC cells to Doxorubicin and Docetaxel, as measured by increased apoptosis and cell death. We also show that WAVE3 knockdown in the chemotherapy-treated TNBC cells results in inhibition of STAT1 phosphorylation, as well as a significant decrease in expression levels of its downstream effector Hif-1α. Since Hif-1α is a major activator of VEGF-A production, and therefore, a stimulator of tumor angiogenesis, loss of Hif-1α in the WAVE3-knocdown cells resulted in the inhibition the chemotherapy-mediated VEGF-A secretion and the downstream activation of angiogenesis, a phenomenon that often accompanies chemoresistance. Our data identify a critical role of WAVE3 in sensitizing TNBC to chemotherapy by inhibiting the STAT1⌠Hif-1α⌠VEGF-A signaling axis, and support the possibility of WAVE3 inhibition as a promising approach for TNBC cancer therapy.

Citation Format: Gangarao Davuluri, Edward F. Plow, William P. Schiemann, Khalid Sossey-Alaoui. WAVE3 is associated with chemoresistance in TNBC by activating tumor angiogenesis downstream of the STAT1/Hif-1α/VEGF-A signaling axis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2925. doi:10.1158/1538-7445.AM2015-2925
Abstract 2927: Targeting p53 with Bak and Bax mitochondrial targeting signals for the treatment of ovarian cancer
Authors: Lu Phong
Published: August 2015

Although the main function of p53 is a nuclear transcription factor that has important roles in cell cycle arrest, DNA repair, and apoptosis, p53 can directly trigger the intrinsic apoptotic pathway through the mitochondria. p53 has been known to bind to mitochondrial anti-apoptotic proteins (Bcl-2 and Bcl-XL) and pro-apoptotic proteins (Bak and Bax), which will cause the oligomerization of Bak and Bax. The result is the formation of permeable pores on the mitochondrial outer membrane, which in turn cause the release of cytochrome c and activation of caspase-3. Targeting p53 to the mitochondria is an attractive approach because it can cause a rapid apoptotic response. We have introduced the mitochondrial targeting signal (MTS) from Bak or Bax to the C-terminus of p53 and have shown the superior efficacy of p53-BakMTS and p53-BaxMTS over wtp53 in many human cancer cell lines. We have identified that the DNA binding domain (DBD) of p53 may be the minimal domain of p53 required for apoptosis. Our preliminary data in SKOV-3 ovarian cancer cells also suggests that DBD-BakMTS may work just as well as full length p53-BakMTS. Further in humans, the single nucleotide polymorphism at codon 72 from Pro to Arg is known to enhance the mitochondrial targeting of p53. We have recently made the P72R mutation in our p53 constructs. Our goal is to use a novel version of p53 directed to the mitochondria as a direct apoptogen to treat ovarian cancer, the most lethal gynecological malignancy with 69% of patients succumbing to this disease. Our next step will be to test these constructs in vitro in human ovarian cancer cell lines and in vivo using the syngeneic ID8 mouse model. 7-AAD cell death assay was performed in the mouse ovarian ID8 cells transfected with our constructs. p53-Bax-MTS and p53-BakMTS appear to be the two most effective constructs. Interestingly, full length p53 is required for its apoptotic activity in this ID8 mouse cell line. In terms of the P72R polymorphism, p53(72R)-BakMTS and p53(72R)-BaxMTS do not show enhanced apoptotic potential in the ID8 mouse line, most likely due to the sequence differences between human and mouse p53. However, it is possible that the combination of 72R polymorphism with MTS from Bak and Bax may produce a superior mitochondrial localization of p53 in human cell lines. Our ultimate goal is to develop mitochondrially targeted p53 constructs that directly induce apoptosis for the treatment of ovarian cancer.

Acknowledgements: NIH CA151847

Citation Format: Phong Lu. Targeting p53 with Bak and Bax mitochondrial targeting signals for the treatment of ovarian cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2927. doi:10.1158/1538-7445.AM2015-2927
Abstract 2975: Comprehensive analysis of genetic alterations and DNA copy number in Borrmann type IV gastric cancer
Authors: Yasumoto Makiko , Ogasawara Sachiko , Akiba Jun , Kusano Hironori , Sumi Akiko , Isobe Taro , Kizaki Junya , Akagi Yoshito , Torimura Takuji , Sakamoto Etsuko , Itadani Hiraku , Kobayashi Tsutomu , Mizuarai Shinji , Oie Shinji , Yano Hirohisa
Published: August 2015

The prognosis of patients with Borrmann type IV gastric cancer (Type IV) is extremely poor. Thus, there is an urgent need to elucidate the molecular mechanisms underlying the oncogenesis of Type IV and to identify new therapeutic targets. Although previous studies using whole-exome and whole-genome sequencing have elucidated genomic alternations in gastric cancer, no reports have focused on the comprehensive genetic analysis in Type IV. To find cancer relevant genes in Type IV, we performed whole exome sequencing and genome-wide copy number analysis on 14 patients of Type IV. The exome sequencing identified a total of 182 somatic mutations which cause amino-acid substitutions or splice-site alterations. Previously reported mutations in gastric cancer were detected, such as ARID1A, CDH1, TP53 and RHOA. Moreover, our study has found novel mutations that have not been reported in gastric cancer and other tumor types. Among the newly identified genetic alterations, a mutation in muscle RAS oncogene homolog (MRAS) was predicted to cause molecular dysfunction. MRAS belongs to the Ras subgroup of small G proteins, including the prototypic RAS oncogene, KRAS, HRAS, and NRAS. The MRAS mutation discovered by whole-exome sequencing was validated by mutation-specific PCR. To investigate the frequency of mutation in MRAS, we performed Sanger sequencing of additional 46 Type IV samples. A total of 9 non-synonymous mutations were discovered with a mutation frequency of 17%, showing that MRAS is recurrently mutated in Type IV. The genome-wide copy number analysis also showed that copy number was aberrant in 41 genes, including Insulin-like growth factor 1 receptor (IGF1R). The samples with IGF1R amplification showed remarkably higher expression levels of mRNA and protein compared with other samples. In conclusion, this is the first research to discover mutation in MRAS among human tumor samples, although introduction of artificially mutated MRAS is known to have oncogenic activity. In addition, these results suggested that MRAS mutation and IGF1R amplification could drive tumorigenesis of Type IV and could be a new therapeutic target for Type IV.

Citation Format: Makiko Yasumoto, Sachiko Ogasawara, Jun Akiba, Hironori Kusano, Akiko Sumi, Taro Isobe, Junya Kizaki, Yoshito Akagi, Takuji Torimura, Etsuko Sakamoto, Hiraku Itadani, Tsutomu Kobayashi, Shinji Mizuarai, Shinji Oie, Hirohisa Yano. Comprehensive analysis of genetic alterations and DNA copy number in Borrmann type IV gastric cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2975. doi:10.1158/1538-7445.AM2015-2975
Abstract 298: Overcoming TGF-β-related immunosuppression for cancer immunotherapy by oncolytic adenovirus co-expressing Interleukin-12 and decorin
Authors: Oh Eonju , Choi Il-Kyu , Hwang June Kyu , Yun Chae-Ok
Published: August 2015

Interleukin (IL)-12 has been shown to be one of the most promising antitumor cytokines. The major antitumor activities of IL-12 rely on its ability to promote T helper type 1 (Th1) immune responses and CTL responses. However, immune suppression-dominated microenvironments in advanced tumors hinder the effect of IL-12 gene therapy for cancer. Most tumor cells produce large amounts of transforming growth factor-β (TGF-β), which plays crucial roles of tumor-induced immune suppression. The TGF-β inhibits proliferation of immune cells such as T, NK, and dendritic cells, resulting in suppression of antitumor immunity. Moreover, TGF-β is also known to regulate the maintenance and induction of regulatory T (Treg) cells. Thus, it has been hypothesized that inhibiting TGF-β might be an apt strategy for generating more effective cancer immunotherapy. Therefore, to enhance the efficiency of IL-12-mediated cancer immunotherapy, we generated oncolytic adenovirus co-expressing IL-12 and Decorin (hDCN) (RdB/IL12/hDCN) as suitable therapeutic adjuvant to overcome immune suppression by down-regulation of TGF-β through hDCN. Intratumoral administration of RdB/IL12/hDCN promoted enhanced antitumor effects compared with an oncolytic adenovirus expressing IL-12 or hDCN alone (RdB/IL12 or RdB/hDCN, respectively). We showed markedly elevated levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α and IFN-γ secreting cells in RdB/IL12/hDCN-treated tumors. Moreover, Treg in draining lymph nodes (DLNs) dramatically decreased in mice treated with RdB/IL12/hDCN, suggesting an association with the down-regulation of TGF-β in tumor treated with RdB/IL12/hDCN. Consistent with these data, tumor tissue injected with RdB/IL12/hDCN showed increased infiltration of CD4+ T and CD8+ T cells infiltration. Furthermore, hDCN enhanced viral distribution and tumor penetration, leading to improved virus-mediated cancer gene therapy by overcoming the extracellular matrix barrier within tumor masses. Together, these results provide new insight into the ability of hDCN to eliminate immune suppression and demonstrate that adenovirus co-expressing IL-12 and hDCN is a promising therapeutic tool for cancer treatment than adenovirus expressing IL-12 alone.

Note: This abstract was not presented at the meeting.

Citation Format: Eonju Oh, Il-Kyu Choi, June Kyu Hwang, Chae-Ok Yun. Overcoming TGF-β-related immunosuppression for cancer immunotherapy by oncolytic adenovirus co-expressing Interleukin-12 and decorin. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 298. doi:10.1158/1538-7445.AM2015-298
Abstract 303: Effect of scaffold on drug sensitivity of multicellular spheroids: Which method is close to in vivo and suitable for HTS
Authors: Masuda Norio , Mizuno Atsushi , Rahman M.Mamunur , Arai Kazuya , Itoh Manabu
Published: August 2015

Background: Three-dimensional (3D) cell culture models are more extensive to be better models than two-dimensional (2D) cell culture models due to enriched cellular signaling pathways, cell to-cell contact, more representative of tissue morphology, and predictive of drug responses in vivo. Different drugs screening and various biological responses between 2D and 3D cell culture models have been reported. However, very little information is available on cell function and/or drug susceptibility caused by a difference in methodology of three dimensional culture. Here, we compared drug responses of various cancer cells against distinctive anti-cancer drugs when grown in monolayer, scaffold 3D culture (Matrigel, NanoCuluture Plate (NCP)) or scaffold-free 3D culture models (ultra-low attachment round bottom plate), and also confirmed which method is suitable for high-throughput screening for robust three-dimensional screening model.

Method: We examined drug sensitivities of different lung cancer (A549, H226, H1650), breast cancer (SKBR-3, JIMT1), and prostate cancer (DU145) cell lines under monolayer or 3D cell culture conditions against several anti-cancer agents i.e. molecular targeted agent (Gefitinib, Afatinib, Temsirolimus, Vemurafenib, Vismodegib, Sorafenib), demethylation agent (Azacytidine, Decitabine), anti-microtubule agent (Docetaxel), alkylating agent (Cisplatin, Irinotecan), antimetabolite agent (5-FU, Gemcitabine, Methotrexate). The conditions of these assays are optimized for each culture method. Cell proliferation was measured by ATP assay. Furthermore, we analyzed the biological/ morphological differences among 3D culture methods (scaffold or scaffold-free culture models).

Result: Large variations in drug responses were observed among the different cell culture models. Cells grown in 3D scaffold culture model (NCP) were more drug sensitive than other cell culture models. Especially docetaxel, 5-FU, and methotrexate showed clear growth inhibition in three dimensional scaffold culture models (NCP) than scaffold-free culture models (round-bottom).

Conclusion: In this study, we showed that scaffold 3D culture is appropriate for anti-cancer drug-susceptibility test. We also proposed that NanoCuluture plate which is 3D scaffold culture model, can be used as a sophisticated high-throughput three-dimensional drug screening model.

Citation Format: Norio Masuda, Atsushi Mizuno, M.Mamunur Rahman, Kazuya Arai, Manabu Itoh. Effect of scaffold on drug sensitivity of multicellular spheroids: Which method is close to in vivo and suitable for HTS. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 303. doi:10.1158/1538-7445.AM2015-303
Abstract 3046: Glutamate dehydrogenase 1 signals through antioxidant glutathione peroxidase 1 to regulate redox homeostasis and tumor growth
Authors: Jin Lingtao , Li Dan , Alesi Gina , Fan Jun , Kang Hee-Bum , Zhou Lu , Boggon Titus , Magliocca Kelly , He Chuan , Arellano Martha , Khoury Hanna , Shin Dong , Khuri Fadlo , Kang Sumin
Published: August 2015

How mitochondrial glutaminolysis contributes to redox homeostasis in cancer cells remains unclear. Here we report that the mitochondrial enzyme glutamate dehydrogenase 1 (GDH1), commonly upregulated in human cancers, is predominantly important for redox homeostasis in cancer cells by controlling the intracellular levels of its product alpha-ketoglutarate (a-KG) and subsequent metabolite fumarate. Mechanistically, fumarate binds to and activates a ROS scavenging enzyme glutathione peroxidase 1 (GPx1) to regulate redox homeostasis, which provides a proliferative advantage to cancer cells and tumor growth. Our findings not only provide novel insights into understanding of the role of glutaminolysis in redox homeostasis but also suggest a novel signaling function of fumarate that regulates GPx1, allowing additional crosstalk between glutaminolysis, TCA cycle and redox status. Targeting GDH1 by shRNA or a newly identified small molecule inhibitor R162 resulted in imbalanced redox homeostasis, leading to attenuated cancer cell proliferation and tumor growth. Thus, our findings provide proof-of-principle suggesting GDH1 as a promising therapeutic target in the treatment of human cancers associated with elevated glutamine metabolism.

Citation Format: Lingtao Jin, Dan Li, Gina Alesi, Jun Fan, Hee-Bum Kang, Lu Zhou, Titus Boggon, Kelly Magliocca, Chuan He, Martha Arellano, Hanna Khoury, Dong Shin, Fadlo Khuri, Sumin Kang. Glutamate dehydrogenase 1 signals through antioxidant glutathione peroxidase 1 to regulate redox homeostasis and tumor growth. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3046. doi:10.1158/1538-7445.AM2015-3046
Abstract 3048: Study of mitochondria in multi-drug resistance and Polyphyllin D anti-cancer effect in hepatocellular carcinoma
Authors: Ngai Brandon Ho , Loo Jacky Fong Chuen , Kong Siu Kai
Published: August 2015

Liver cancer, being the sixth common cancer, causes approximately 700,000 deaths worldwide annually. Major treatment relies on chemotherapies because of the late diagnosis in many cases of liver cancer. However, the results are not satisfactory since the cancer cells develop multi-drug resistance (MDR), which greatly hampers the efficacy of multiple kinds of structurally different drugs, during the course of treatment. Several mechanisms of MDR were identified, the most notable one is the over-expression of ATP-binding cassette (ABC) transporters, such as P-glycoprotein (P-gp) and multi-drug resistance protein 1 (MRP-1), on plasma membrane. These transporters act as efflux pump of various types of chemo-drugs to accentuate the MDR phenotype. Nevertheless, the complete picture of MDR formation is yet to be explored. As mitochondria contribute vitally in apoptosis and energy production, in which both criteria are substantially altered in cancer context, we hypothesized that the organelle also takes part in the development of MDR in liver cancer. In our study, with the support of conventional assays such as western blotting and confocal imaging analysis, we adopted a novel approach by making use of the tunable resistive pulse sensing (TRPS) technique to characterize isolated mitochondria of HepG2 and R-HepG2, which corresponds to the native and multi-drug resistant liver cancer cell lines respectively. We identified a difference in volume and amount of mitochondria present in HepG2 and R-HepG2, plus the expression of P-gp and MRP-1 on mitochondria of R-HepG2. These suggest a potential role of mitochondria in MDR. On the other hand, a drug candidate Polyphyllin D (PD) was found to exert selective potency against R-HepG2. However, the mechanism is not lucid. We investigated the possible participation of mitochondria in this phenomenon and identified PD causes stronger swelling and membrane potential depolarization to isolated mitochondria of R-HepG2. Future attempt in elucidating pathways involved in PD toxicity will bring insight on designing therapeutics against cancer cells with MDR phenotype. This study also suggested future research in mitochondria can aid the discovery of more mechanisms in MDR development.

Citation Format: Brandon Ho Ngai, Jacky Fong Chuen Loo, Siu Kai Kong. Study of mitochondria in multi-drug resistance and Polyphyllin D anti-cancer effect in hepatocellular carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3048. doi:10.1158/1538-7445.AM2015-3048
Abstract 3049: BRAF status and low glutamine/glucose influence glutamine addiction modulated by oxidative stress in human melanoma cells
Authors: Rieber Manuel , Chavez-Perez Valery , Strasberg-Rieber Mary
Published: August 2015

BACKGROUND.- Cancer cells from solid tumors become metabolically stressed, when nutrients are insufficient within poorly vascularized regions. Metabolic stress results from severe deprivation of oxygen, glutamine and glucose, partly through excessive reactive oxygen species (ROS) production. Mutant BRAF can stimulate the accumulation of hypoxia-inducible factor-1alpha (HIF-1α) in physiological glucose and supplementation with supra-physiological glutamine in melanoma cells (Kumar SM et al, Cancer Res. 2007;67:3177-84).Although HIF1α reprograms melanomas to become highly glycolytic, this can be impaired in a microenvironment with sub-physiological levels of glutamine and glucose (Kwon SJ, Lee YJ.Clin Cancer Res. 2005;11:4694-700).,because melanomas require glycolysis and mitochondrial glutamine utilization. PURPOSE.- To compare proliferation and survival in response to glutamine and glucose restriction in wt-BRAF or mutant BRAF melanoma. EXPERIMENTAL. Determination of metabolic activity/proliferation and cell survival by fluorometric assays and crystal violet staining. RESULTS.- Glutamine depletion diminished survival even in physiological 5 mM glucose, particularly during hypoxia. However, 2.5 mM glucose or exogenous pyruvate when added together with another anti-oxidant N-acetylcysteine (NAC) partly substituted for glutamine in normoxic wt-BRAF cells. A much lower attenuation was seen in glutamine-depleted mutant BRAF cells when supplemented with NAC and pyruvate, particularly, under hypoxia. CONCLUSIONS.-1) glutamine restriction decreases metabolic activity and tumor cell survival, irrespective of BRAF status, being more toxic to mut-BRAF cells, either under hypoxia or in physiological glucose and normoxia,. 2) Antagonizing glutamine addiction may decrease tumor recurrence if combined with anti-BRAF targeted therapy.

Citation Format: Manuel Rieber, Valery Chavez-Perez, Mary Strasberg-Rieber. BRAF status and low glutamine/glucose influence glutamine addiction modulated by oxidative stress in human melanoma cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3049. doi:10.1158/1538-7445.AM2015-3049
Abstract 3055: Regulation of mitochondria-nuclear crosstalk in triple negative breast cancer
Authors: Park Jun H. , Vithayathil Sajna A. , Putluri Nagireddy , Tsouko Efrosini , Donti Taraka R. , Frigo Daniel E. , Creighton Chad J. , Lewis Michael T. , Sreekumar Arun , Wong Lee-Jun , Kaipparettu Benny A.
Published: August 2015

Driver pathways of triple negative breast cancer (TN BCa) is still poorly understood. Thus, it is important to identify the underlying mechanisms of TN BCa progression. Importantly, African American and Hispanic patients are more likely to be diagnosed of BCa at a younger age, have a higher probability of developing TN BCa. They also diagnosed with a more advanced stage of the disease compared to Caucasian women.

Src oncognenic pathway is one of the major tumor driving mechanisms in TN BCa. Src pathway is frequently over-expressed in TN BCa and Src inhibitors are one of their current treatments of choices. However, recent single drug therapy Phase-II trials with Src inhibitors failed to control unselected metastatic TN BCa. This raises some important questions on the underlying mechanism in the regulation of Src in TN BCa especially in aggressive metastatic BCa. Mitochondria, a semiautonomous organelle in cells, play an important role in cellular energy metabolism, free radical generation, and apoptosis. Mitochondria-nuclear crosstalk is a bidirectional pathway of communication between mitochondria and nucleus that influences many cellular and organismal activities. This crosstalk can regulate several oncogenic pathways involved in tumorigenesis. Using transmitochondrial cybrid (cybrid) technology, we generated different cybrid models under common nuclear background of TN BCa. Mitochondria from cells of different cancer potential including benign breast epithelium, moderately and highly metastatic breast cancer cell lines were used to understand cancer mitochondria regulated tumor pathways. Extensive analysis of cybrid models as well as confirmation in parental cell lines and other tumor models suggested that autophosphorylation of c-Src is regulated by mitochondrial tumor characteristics.

Ongoing studies using knock-down and over expression approaches focus on the role of specific mitochondrial proteins that are responsible for mitochondria-nuclear crosstalk and the regulation oncopathways in TN BCa.

Citation Format: Jun H. Park, Sajna A. Vithayathil, Nagireddy Putluri, Efrosini Tsouko, Taraka R. Donti, Daniel E. Frigo, Chad J. Creighton, Michael T. Lewis, Arun Sreekumar, Lee-Jun Wong, Benny A. Kaipparettu. Regulation of mitochondria-nuclear crosstalk in triple negative breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3055. doi:10.1158/1538-7445.AM2015-3055
Abstract 3060: The role of miR-135b in clinical outcome and metastasis in colorectal cancer
Authors: Wang Wei-Ran , Cheng Yawen , Hsu Nan-Yung Hsu
Published: August 2015

Colorectal cancer (CRC) is one of the leading causes of cancer death in both the Western world and Taiwan. Age, gender, and genes are important risk factors for colorectal cancer. Activation of Wnt signaling pathway is an important step involved in the pathogenesis of colorectal cancer. Approximately 70-80% of sporadic colorectal carcinomas have somatic mutations that inactivate APC. But, only 30% CRC patients in Taiwan have APC gene mutation that was significantly lower than the other countries. Thus, the other mechanism involved in APC gene inactivation may contributed to CRC progression in Taiwan. Here, we focus on the Wnt signaling associate microRNAs. Four wnt signaling-associated microRNAs were selected in this study, including miR-21, miR-200a, miR-135a and miR-135b. A total of 156 colorectal cancer patients were enrolled in this study. The expression of miR-21, miR-200a, miR-135a, miR-135b and wnt signaling association genes were analyzed by quantitative RT-PCR and immunohistochemistry. The expression levels of miR-21, miR-200a and miR-135a were not correlated with clinical parameters of CRC patients. The expression of miR-135b were positive correlated with gender, tumor stage, and lymph node metastasis. No any significantly correlation were found between miR-135b, Wnt, β-catenin and APC. But the expression levels of miR-135b were positive correlated with the upstream gene STAT3, β-catenin and C-Myc. Thus, we suggested that upregulation of miR-135b may through β-catenin mediated STAT3 activation.

Citation Format: Wei-Ran Wang, Yawen Cheng, Nan-Yung Hsu Hsu. The role of miR-135b in clinical outcome and metastasis in colorectal cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3060. doi:10.1158/1538-7445.AM2015-3060
Abstract 3107: MicroRNA-1289 is a novel tumor suppressive microRNA in human oral squamous cell carcinoma
Authors: Iwamoto Kazuki , Nakashiro Koh-ichi , Tnaka Hiroshi , Tokuzen Norihiko , Hamakawa Hiroyuki
Published: August 2015

MicroRNAs (miRNAs) are small non-coding double-stranded RNA with sizes of approximately 22 nucleotides, and those inhibit protein translation by binding the 3′-untranslated region of target mRNAs. Each miRNA generally can regulate multiple mRNAs and each mRNA can be inhibited by a number of miRNAs. MiRNAs can behave as not only oncogenes but also tumor suppressive (TS) genes in human malignancies. In fact, a number of TS miRNAs (TS-miRs) were found in human many types of cancer, such as let-7, miR-1, miR-34, miR-145, miR-200, miR-206 and more. In this study, we have attempted to identify a novel TS-miR in human oral squamous cell carcinoma (OSCC) using function-based screening. First, we transfected human OSCC cells (GFP-SAS) with approximately 1,000 synthetic mimicking human mature miRNAs at the concentration of 20 nM complexed with Lipofectamine RNAiMAX. After transfection for 72 hours, the growth of these cells was evaluated by WST-8 assay. As a result, miR-1289 had the most potent growth inhibitory effect against GFP-SAS cells. The growth rate of these cells transfected with miR-1289 was reduced by 86.4%. MiR-1289 also remarkably inhibited the growth of other human OSCC cells (Ca9-22, HSC2, HSC3 and primary cultured cells). Subsequently, we assessed the growth inhibitory effect of miR-1289 in vivo using a mouse model. GFP-SAS cells (2 × 106) complexed with matrigel in 100 μl aliquots were injected subcutaneously in nude mice. After tumor formation, we administered 100 ng of miRNA with 0.5% atelocollagen complexes (100 μl) around tumor every 3 days for a total of 5 injections. MiR-1289/atelocollagen complex administration group significantly reduced the size of subcutaneously xenografted GFP-SAS tumors compared with the control group. Next, we investigated the expression of miR-1289 in OSCC tissues by real-time quantitative RT-PCR. The expression levels of miR-1289 were significant decreased in OSCC tissues compared to adjacent normal oral mucosa tissues. Finally, target genes of miR-1289 were explored by microarray and Ingenuity Pathway Analysis (IPA). Microarray analysis revealed that the expression levels of 51 genes were commonly down-regulated in OSCC cells (GFP-SAS, Ca9-22 and HSC2) by transfection with miR-1289. Among these genes, 15 genes were identified as target genes of miR-1289 by IPA. These results suggest that miR-1289 functions as a novel TS-miR in OSCC, and may be a useful therapeutic tool for the patients with OSCC.

Citation Format: Kazuki Iwamoto, Koh-ichi Nakashiro, Hiroshi Tnaka, Norihiko Tokuzen, Hiroyuki Hamakawa. MicroRNA-1289 is a novel tumor suppressive microRNA in human oral squamous cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3107. doi:10.1158/1538-7445.AM2015-3107
Abstract 3127: Small but lethal, miR-137 acts as a tumor suppressive microRNA in colorectal cancer
Authors: Smith Amber Rae , Marquez Rebecca , Tsao Bryan , Roy Alexandria , Wilkerson Bailey , Pathak Surajit , Neufeld Kristi , Sun Xiao-Feng , Xu Liang
Published: August 2015

Colorectal cancer is the 3rd most common cause of cancer-related deaths for men and women in the United States. Understanding the mechanisms associated with the initiation and progression of colorectal cancer is needed to develop precise therapies. Multiple regulatory processes are responsible for maintaining cellular homeostasis, including microRNA (miRNA) regulation. We previously demonstrated that miR-137 expression was significantly decreased in 84% of tumors as compared to adjacent mucosal tissue from patients with rectal cancer (n = 68). Restoration of miR-137 reduced colon cancer growth, measured by colony formation and tumorsphere assays. Induction of miR-137 expression significantly decreased tumor growth in our xenograft tumor model by approximately 50% (n = 10). To understand the mechanism associated with the tumor suppressive function of miR-137, we transfected miR-137 mimic into HCT-116 cells and performed microarray analysis to identify changes in gene expression and identify key regulatory pathways effected by miR-137 restoration. Multiple miR-137 targets were down regulated, as compared to cells treated with a negative control mimic. The data was validated using quantitative real-time PCR (qPCR). Finally, the expression of miR-137 target genes was analyzed in a panel of colon cancer cell lines using qPCR. In summary, miR-137 acts as a tumor suppressive microRNA by negatively regulating multiple targets that are overexpressed in colorectal cancer.

Citation Format: Amber Rae Smith, Rebecca Marquez, Bryan Tsao, Alexandria Roy, Bailey Wilkerson, Surajit Pathak, Kristi Neufeld, Xiao-Feng Sun, Liang Xu. Small but lethal, miR-137 acts as a tumor suppressive microRNA in colorectal cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3127. doi:10.1158/1538-7445.AM2015-3127
Abstract 3146: T cell therapy targeting a neoantigen reduces in vivo tumour growth
Authors: Inderberg-Suso Else M. , Wälchli Sebastien , Myhre Marit R. , Wang Mengyu , Almåsbak Hilde , Kvalheim Gunnar , Gaudernack Gustav
Published: August 2015

T-cell receptor (TCR) transfer is an attractive strategy to increase the number of cancer-specific T cells in adoptive cell therapy. However, recent clinical and pre-clinical findings indicate that careful consideration of the target antigen is required to avoid on-target, off-site toxicity. A safer alternative to targeting tumour-associated self-antigens may be to direct engineered T cells against mutated proteins such as frequently occurring frameshift mutations. Furthermore, such frameshift mutations result in novel polypeptides allowing selection of TCRs from the non-tolerant T-cell repertoire circumventing the problem of low affinity TCRs due to central tolerance. The transforming growth factor β Receptor II frameshift mutation (TGFβRIImut) is found in Lynch syndrome cancer patients and in around 15% of sporadic colorectal and gastric cancers displaying microsatellite instability.

The -1A mutation within a stretch of 10 adenine bases (nucleotides 709-718) of the TGFβRII gene gives rise to immunogenic peptides previously used for vaccination of MSI+ colorectal cancer patients in a Phase I clinical trial. From a clinically responding patient we isolated a CD8neg/CD4neg CTL clone from which we cloned an HLA-A2-restricted TGFβRIImut-specific TCR. We showed that both CD8+ and CD4+ T cells transiently redirected with this TGFβRIImut-TCR recognised colon carcinoma cell lines harbouring the frameshift mutation, indicating CD8+ co-receptor independency.

Cysteine modification of the TCR enhanced its expression and function and we have demonstrated that these TCR-redirected T cells significantly enhanced survival and reduced tumour growth in a NOD/SCID xenograft mouse model of colorectal cancer.

Citation Format: Else M. Inderberg-Suso, Sebastien Wälchli, Marit R. Myhre, Mengyu Wang, Hilde Almåsbak, Gunnar Kvalheim, Gustav Gaudernack. T cell therapy targeting a neoantigen reduces in vivo tumour growth. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3146. doi:10.1158/1538-7445.AM2015-3146
Abstract 3151: T cells engineered to express a chimeric antigen receptor targeting chondroitin sulfate proteoglycan 4 (CSPG4) specifically kill medulloblastoma and produce inflammatory cytokines
Authors: Rota Christopher M. , Tschernia Nicholas , Feldman Steven , Mackall Crystal , Lee Daniel W.
Published: August 2015

Human T-cells expressing chimeric antigen receptors (CARs) recognizing common cancer antigens have shown great promise in the clinic with the potential for significant anti-tumor responses. CSPG4 is a chondroitin sulfate proteoglycan previously shown to be overexpressed in melanoma and breast cancer, as well as glioblastoma stem cells. CARs directed against CSPG4 have recently shown promising pre-clinical efficacy against these tumors. We evaluated the feasibility of targeting CSPG4 using CAR T cell therapy in medulloblastoma and neuroblastoma. Response to standard therapies for advanced neuroblastoma and medulloblastoma is poor and carries significant toxicities, which may be ameliorated by immunotherapy. We found that multiple medulloblastoma and neuroblastoma cell lines (DAOY, D283 Med, D425 Med, UW-228, and SKNAS) express CSPG4 at high levels, as does 143b osteosarcoma as a positive control. By contrast, CSPG4 in normal tissue is expressed only on pericytes and in low levels on small bowel as previously assessed by protein array [Beard et al, J Immunother Cancer 2014]. To evaluate therapeutic efficacy, a second generation CAR with a CD3ζ signaling domain and CD28 co-stimulatory domain containing the TP41.2 anti-CSPG4 scFv was transduced using a γ-retrovirus into T cells. CAR T cells were found to specifically kill 20% of DAOY, 50% of D283 Med, and 50% of 143b cancer cells at a 20:1 effector-to-target ratio in a 4-hour chromium release cytotoxicity assay, while not affecting CSPG4 negative control SY5Y (neuroblastoma) and DND41 (T cell leukemia) cells. Both CD4 and CD8 CAR T cells were also shown to produce IFNγ after exposure to 143b cells. Cytokine production in response to other CSPG4+ lines is ongoing as are cytotoxicity studies with neuroblastomas. For our in vivo model, we have begun to evaluate the efficacy of CAR T cells delivered either intravenously or via intratumoral injection against DAOY and D283 Med cells that have been orthotopically implanted in the cerebellum of immunodeficient NOD/SCID/IL2Rγ-/- mice. These results, if successful, will inform the development of a new clinical protocol for the treatment of children and young adults with medulloblastoma and neuroblastoma.

Citation Format: Christopher M. Rota, Nicholas Tschernia, Steven Feldman, Crystal Mackall, Daniel W. Lee. T cells engineered to express a chimeric antigen receptor targeting chondroitin sulfate proteoglycan 4 (CSPG4) specifically kill medulloblastoma and produce inflammatory cytokines. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3151. doi:10.1158/1538-7445.AM2015-3151
Abstract 3169: Influence of systemic and local immune response on overall survival after hepatic resection of colorectal liver metastasis
Authors: Hamada Takeomi , Hidenobu Ishizaki , Haruyama Yukihiro , Hamada Rouko , Yano Koichi , Otani Kazuhiro , Fujii Yoshiro , Kondo Kazuhiro , Kataoka Hiroaki
Published: August 2015

Background: Colorectal cancer progression is dependent on complex interactions between the tumor and the host. Tumor infiltrating lymphocytes (TIL) have been reported as a prognostic factor in colorectal cancer. The aim of this study was to assess the prognostic value of pre-operative systemic immune biomarkers and TIL in patients with resectable colorectal liver metastases (CLM).

Methods: Clinicopathological data were reviewed for patients (n = 31) underwent hepatic resections for CLM between 2000 and 2008. Prognostic variables are factors of the primary lesion, factors of the metastatic lesion, and factors of host inflammatory biomarkers measured from routine pre-operative blood tests (modified Glasgow Prognostic Score and neutrophil to lymphocyte ratio (NLR)). Univariate and multivariate analyses for prognostic factors were performed. To evaluate the local immune responses for resected metastatic lesion, immunohistochemical analysis was performed. The immune subsets were CD8+ T cells, CD45RO+ T cells, Fork head box p3+ (FOXP3+) T cells, and CD66b+ neutrophils.

Results: The number of metastases ≥ 4, extrahepatic metastasis, adjuvant chemotherapy, and raised NLR > 4.1 were correlated with cancer-specific survival in univariate analysis. On multivariate analysis, the number of metastases ≥ 4, adjuvant chemotherapy, and elevated NLR remained independent prognostic indicators (metastases ≥ 4: HR = 0.09, CI = 0.01-0.56, p = 0.0102, adjuvant chemotherapy: HR = 0.08, CI = 0.01-0.39, p = 0.0025, NLR: HR = 0.11, CI = 0.01-0.99, p = 0.0494). Accumulation of CD45RO+ T cells and FOXP3+ T cells in the metasatic tumors were significantly associated with favorable survival in patients with resectable CLM (P = 0.02 and 0.04, respectively).

Conclusions: The results of the present study show that the NLR has prognostic value in patients who conduct a hepatic resection for CLM. Moreover, accumulation of CD45RO+ T cells and FOXP3+ T cells in metastatic lesions are independent favorable prognostic factor. Based on the present results and previously literature, systemic immune response including NLR and local immune response may be promising as prognostic factor of patients with resectable CLM.

Citation Format: Takeomi Hamada, Ishizaki Hidenobu, Yukihiro Haruyama, Rouko Hamada, Koichi Yano, Kazuhiro Otani, Yoshiro Fujii, Kazuhiro Kondo, Hiroaki Kataoka. Influence of systemic and local immune response on overall survival after hepatic resection of colorectal liver metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3169. doi:10.1158/1538-7445.AM2015-3169
Abstract 3198: The impact of aerobic exercise on oxygenation and vascularity in breast cancer models
Authors: Wiggins Jennifer M. , Lee Jennifer A. , Rice Lori , Siemann Dietmar
Published: August 2015

Background: Intratumoral hypoxia (pO2 <10mmHg) is associated with aggressive tumor progression and poor patient outcome. It is often found in solid tumors, and can constitute a major obstacle to anticancer therapy. The primary cause of abnormal oxygen tensions in tumors is the aberrant tumor vasculature, characterized by tortuous, leaky and immature vessels. A number of intervention strategies designed to overcome tumor hypoxia have been investigated including high oxygen content breathing and the use of bioreductive prodrugs, but these have had only moderate success in the clinic.

Purpose: The goal of the present investigations was to determine whether aerobic exercise could be applied as a means to improve tumor physiology and decrease tumor hypoxia in breast cancer models.

Methods: The effects of single and repeated bouts of moderate intensity exercise (treadmill running) were studied in mice bearing syngeneic murine mammary carcinoma models (EMT6 and 4T1). The exercise intensities were determined by measuring the anaerobic threshold, which is reflected by the steady rise in blood lactate during an exercise bout. Mice were orthotopically injected with tumor cells and once the tumors reached a size of 250 mm3, the mice were exposed to either a single bout of exercise or daily exercise bouts performed for 5 consecutive days. Controls for each treatment consisted of sedentary mice exposed to a stationary treadmill for the equivalent amount of time. Following cessation of each exercise regime tumors were harvested, sectioned and evaluated by immunofluorescence. Vessel density (MECA-32), blood perfusion (Hoechst-33342) and hypoxia (EF5) were quantified and significance was established at p<0.05.

Results: Preliminary data indicate that even a single exercise bout may begin to demonstrate improvements in blood vessel perfusion and tumor oxygenation. Importantly, after 5 daily bouts of exercise, there was a significant increase in the number of patent vessels and improvement in oxygenation in the tumors of exercising mice compared to those of sedentary controls.

Conclusion: The results indicate that moderate aerobic exercise can improve the oxygenation status of solid tumors. These findings suggest that such a non-toxic intervention may have potential utility in overcoming hypoxia-associated treatment resistance in cancer patients and improve patient outcome.

Citation Format: Jennifer M. Wiggins, Jennifer A. Lee, Lori Rice, Dietmar Siemann. The impact of aerobic exercise on oxygenation and vascularity in breast cancer models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3198. doi:10.1158/1538-7445.AM2015-3198
Abstract 3214: Genome-wide analysis of stroma-tumor crosstalk pathways in lung cancer: therapeutic implications
Authors: Choi Hyejin , Shen Jianting , Durrans Anna , El Rayes Tina , Fischer Kari , Gao Dingcheng , Wong Steve , Altorki Nasser , Mittal Vivek
Published: August 2015

We have systematically dissected the microenvironmental hetereogeneity of lung adenocarcinomas, and provide cellular and molecular landscapes that contribute to carcinogenesis. RNA deep sequencing analysis of individually sorted stromal and tumor epithelial compartments from fresh clinical specimens and a mouse model of KrasG12D; p53-/- tumors identified cell type-specific differentially regulated genes, indicative of tumor “activated/reprogrammed” stroma. We developed a computational crosstalk discovery algorithm to model crosstalk signaling discovery based on ligand-receptor interactions and downstream signaling networks, and identified novel tumor-stroma paracrine and tumor autocrine crosstalk signaling pathways in the lung cancer microenvironment. We have explored the mechanistic basis of selected crosstalk pathways including HGF-MET, IL6-Stat3, Tsp-1-CD36, MMP14-HB-EGF in conferring lung adenocarcinoma progression and in mediating therapeutic resistance, and have used both genetic and pharmacological approaches to block these pathways. This study has the potential for development of therapeutic strategies that target tumor-stroma interactions, and may complement conventional anti-cancer treatments.

Citation Format: Hyejin Choi, Jianting Shen, Anna Durrans, Tina El Rayes, Kari Fischer, Dingcheng Gao, Steve Wong, Nasser Altorki, Vivek Mittal. Genome-wide analysis of stroma-tumor crosstalk pathways in lung cancer: therapeutic implications. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3214. doi:10.1158/1538-7445.AM2015-3214