bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5729: Loss of USP10 and p14ARF protein expression is associated with poor prognosis patients with small intestinal adenocarcinoma
Authors: Song Joon Seon , Cho Hanbyoul , Chung Joon-Yong , Hwang Ilseon , Lee Hee Jin , Hong Seung-Mo , Hewitt Stephen M.
Published: July 2017

Ubiquitin Specific Peptidase 10 (USP10) is a member of the ubiquitin-specific protease family and its related pathways are apoptosis, autophagy and DNA damage. P14ARF is encoded by an alternative reading frame within the INK4a/ARF locus at chromosome 9p21, which is related with cell cycle arrest and apoptosis. Recently, ablation of USP10 prevented c-myc induced cellular senescence by destabilization of p14ARF and p53, and accelerated hyper-proliferation and transformation of normal cells, suggesting USP10 have a crucial role in oncogenes induced senescence by keeping the stability of p14ARF and p53, key regulator of cellular senescence. However, knowledge on the clinical and prognostic significance of USP10 and p14ARF expression in patients with small intestine adenocarcinoma is limited. In the present study, we investigated the prognostic significance of USP10 and p14ARF by immunohistochemistry. 195 of resected small intestine adenocarcinomas were retrieved from 22 South Korean institutions by Korean Small Intestinal Cancer Study Group. Tissue microarrays were constructed using formalin-fixed paraffin-embedded specimen. 70 (35.9 %) out of 195 patients showed immunoreactivity for USP10 and 124 (63.6 %) showed loss of immunoreactivity for USP10. 120 (61.5 %) of 195 patients were immunopositive for p14ARF and 75 (38.5 %) patients showed loss of immunoreactivity for p14ARF. Loss of USP 10 was correlated with male predominancy (p=0.014), higher pT stage (p=0.044), presence of lymphatic invasion (p=0.033), absences of sporadic adenoma (p=0.024) and absences of peritumoral dyplasia (p=0.019). On multivariate survival analysis, loss of USP10 expression was independent poor prognostic factor ( HR=3.04, 95% CI= 1.79-5.18, p < 0.001) and also both loss of USP10 and p14ARF expressions showed similar result ( HR=3.76, 95% CI= 1.93-7.23, p < 0.001) while loss of p14ARF was not an independent prognostic factor ( p= 0.587). Our findings indicate that loss of USP10, associated with loss of p14ARF protein expression, could be poor prognostic factors in small intestine adenocarcinoma.

Citation Format: Joon Seon Song, Hanbyoul Cho, Joon-Yong Chung, Ilseon Hwang, Hee Jin Lee, Seung-Mo Hong, Stephen M. Hewitt. Loss of USP10 and p14ARF protein expression is associated with poor prognosis patients with small intestinal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5729. doi:10.1158/1538-7445.AM2017-5729
Abstract 5752: Susceptibility of cells from cervical transformation zone to HPV-16 induced immortalization and dysplastic differentiation
Authors: Deng Han , Hillpot Eric , Yeboah Philomina , Mondal Sumona , Woodworth Craig D.
Published: July 2017

Persistent infection of high-risk human papillomavirus (HPV) is a major risk factor for cervical cancer. Greater than 90% of these cancers originate in the cervical transformation zone (CTZ), a narrow band of squamous metaplastic epithelium between ectocervix and endocervix. It is unclear why the CTZ has high susceptibility to malignant transformation and few studies have specifically examined cells from this region. We hypothesized cells cultured from human CTZ are uniquely susceptible to cellular immortalization and dysplastic differentiation, two alterations that contribute to malignant development. Primary epithelial cells were cultured from each region of human cervix (ectocervix, endocervix and CTZ) and characterized for expression of differentiation markers to assess their origin. We examined susceptibility of cells from each region to immortalization after transfection with the complete HPV-16 genome or infection of HPV-16 E6/E7 retroviruses. In addition, we evaluated the extent of dysplastic epithelial differentiation in organotypic cultures of HPV16-immortalized cell lines derived from each region of cervix. Our results showed that cells from the CTZ express specific keratin markers (keratin 14 and 18) that confirm their origin. Primary cell cultures from CTZ and ectocervix are more susceptible to immortalization by the HPV16 complete genome or E6/E7 retroviruses. In contrast, preliminary results indicate that HPV-immortalized CTZ and endocervical cells are more susceptible to dysplastic differentiation when maintained in organotypic culture. Our results may help to identify important signal pathways or biomarkers that are unique to cells from CTZ and that can be targeted for improved screening, prevention or therapy.

Citation Format: Han Deng, Eric Hillpot, Philomina Yeboah, Sumona Mondal, Craig D. Woodworth. Susceptibility of cells from cervical transformation zone to HPV-16 induced immortalization and dysplastic differentiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5752. doi:10.1158/1538-7445.AM2017-5752
Abstract 5753: Inhibition of CCAR1, a coactivator of β-catenin, suppresses the proliferation and migration of gastric cancer cells
Authors: Chang Te-Sheng , Lu Chung-Kuang , Wei Kuo-Liang , Chen Yi-Hsing , Cheng Ying-Tung , Tung Shui-Yi , Wu Cheng-Shyong , Chiang Ming-Ko
Published: July 2017

Given a current hypothesis that cancer arises from cancer stem cells (CSCs), targeting key signaling pathways that support CSC self-renewal appears to be a useful approach to cancer therapy. Derranged expression of Wnt signaling is among the major pathways implicated in regulating the stenmess properties of a variety of human cancers, including gastric cancer. Cell cycle and apoptosis regulator 1 (CCAR1) is a transcriptional coactivator which has been shown to be a component of Wnt/β-catenin signaling, and plays an important role in transcriptional regulation by β-catenin. However, the function and clinical significance of CCAR1 in gastric cancer have not been elucidated. Here we show the elevated CCAR1 nuclear expression correlates with the occurrence of gastric cancer. In addition, RNAi-mediated CCAR1 reduction not only suppressed the cell growth and increased apoptosis in gastric cell lines AGS and MKN28, but also reduced the migration and invasion ability of these cells. Furthermore, In vivo xenograft assay revealed that the expression level of CCAR1 was critical for tumorigenesis. Our data demonstrates that CCAR1 contributes to carcinogenesis in gastric cancer and is required for the survival of gastric cancer cells. Moreover, CCAR1 may serve as a diagnostic marker and a potential therapeutic target.

Citation Format: Te-Sheng Chang, Chung-Kuang Lu, Kuo-Liang Wei, Yi-Hsing Chen, Ying-Tung Cheng, Shui-Yi Tung, Cheng-Shyong Wu, Ming-Ko Chiang. Inhibition of CCAR1, a coactivator of β-catenin, suppresses the proliferation and migration of gastric cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5753. doi:10.1158/1538-7445.AM2017-5753
Abstract 5757: Study of human mammary tumor virus (HMTV) in human breast cancer by NanoString nCounter and FISH analysis
Authors: Melana Stella M. , Tripodi Joseph , Nosike Digna , Shabnam Jaffer , Etkind Polly , Pogo Beatriz GT , Holland James F.
Published: July 2017

Human Mammary Tumor Virus (HMTV), a retrovirus 90-98% homologous to Mouse Mammary tumor Virus (MMTV), the etiological agent of murine mammary cancer, has been detected by PCR in 40% of American women’s breast cancers, but not in healthy tissues of the same breast (Cancer Research 1995, 55:5173-79; Clin. Cancer Research 2001, 7-283-4). A complete 9.9 kb HMTV proviral sequence has been detected in breast cancer genomic DNA (Cancer Res 2001,61:1754-9). HMTV viral particles with betaretrovirus characteristics have been isolated from metastatic breast cancer cells in effusions (MSSM cells). Expression of HMTV proteins has been detected in MSSM cells by western blot, fluorescence-activated cell sorting analysis, and immunofluorescence assays but not in normal mammary epithelial cells (J Virol Methods 2010, 163(1): 157-61). PCR is a sensitive technique susceptible of contamination, which due to amplification, could result in false positive detection. Although murine DNA was not found by PCR, the possibility still exists that HMTV in human DNA could be a result of laboratory contamination. Despite the substantial evidence supporting the presence of HMTV in breast cancer the controversy continues because the chance of contamination has led to doubt that murine-like viruses are human pathogens. We now report additional analyses of breast cancer specimens employing other methods for HMTV detection that reinforce previous findings. NanoString nCounter, a new technology using specific 100 mer oligonucleotides as probes bound to reporter molecules that can detect HMTV gag, env and LTR sequences as well as two murine specific probes in a single reaction without amplification. Flourescence in situ hybridization (FISH) assays with a HMTV probe is a technology that permits visualization of HMTV proviral DNA in the nuclei of surgical samples of breast cancer as well as in MSSM cells. Results from Nanostring nCounter, and FISH analysis show the presence of HMTV DNA in surgical cancer specimens and in MSSM cells. The data exclude contamination and confirm the authenticity of HMTV in human breast cancer. Next-generation sequencing and other studies are in progress to determine the role of HMTV in the pathogenesis of breast carcinoma.

Citation Format: Stella M. Melana, Joseph Tripodi, Digna Nosike, Jaffer Shabnam, Polly Etkind, Beatriz GT Pogo, James F. Holland. Study of human mammary tumor virus (HMTV) in human breast cancer by NanoString nCounter and FISH analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5757. doi:10.1158/1538-7445.AM2017-5757
Abstract 5764: Oncogenic role of GAEC1 and its potential modulation with p53 in pathogenesis of colon cancer
Authors: Wahab Riajul , Islam Farhadul , Gopalan Vinod , Lam Alfred King-Yin
Published: July 2017

Introduction: GAEC1 (Gene amplified in esophageal cancer 1), is frequently amplified and overexpressed in colon cancer tissues. In the present study, we aimed to unveil the oncogenic potential of GAEC1 in carcinogenesis of colon cancer by studying the underlying cellular functions and molecular interactions by in vitro and in vivo experiments.

Method: Transient overexpression of GAEC1 with pcDNA3.1-GAEC1 and silencing with GAEC1-siRNA was performed and several downstream assays were done such as migration, clonogenic and apoptotic assay. Analysis of cell kinetics was done using flow cytometry and cell counting kit-8 was used for cell proliferation assay. Immunofluorescence and Western blot assay were used to determine the expression of different target proteins. Co-immunoprecipitation was used to confirm the protein-protein interaction. For xenotransplantation the severely combined immunodeficient (SCID) mice (4 groups, 6 in each group) were injected subcutaneously with GAEC1 shRNA and control shRNA transfected (stable) SW480 and SW48 colon cancer cells.

Result: The overexpression of GAEC1 increased cell proliferation, migration, reduced apoptosis in colon cancer cells. Also, these cells showed cell cycle arrest at the synthetic phase, activation of Bcl-2, K-ras, pAKT proteins as well as inhibition of p53, PUMA, p21 and BAX proteins. Conversely, knockdown of GAEC1 reduced cell proliferation, migration, decreased the phosphorylation of AKT, and induced apoptosis, G2/M phase arrest and cleavage of poly (ADP-ribose) polymerase (PARP). Co-immunoprecipitation revealed GAEC1’s interaction with p53. In addition ectopic over expression and silencing of GAEC1 lead to reciprocal effects of p53 protein expression. Moreover, knockdown of GAEC1 reduced the nuclear translocation of murine double minute 2 protein (mdm2) indicating that GAEC1 is responsible for the degradation of p53 through the direct interaction between mdm2-p53 in nucleus. Furthermore, our in vivo data demonstrate that the loss of GAEC1 inhibits the tumor formation in xenograft model.

Conclusion: Collectively our study demonstrates that GAEC1 exhibits the oncogenic role in colon cancer by reducing expression of p53 through protein-protein interaction which leads to the inhibition of PUMA, p21, BAX, and activation of Bcl-2, K-ras and pAKT protein expression.

Note: This abstract was not presented at the meeting.

Citation Format: Riajul Wahab, Farhadul Islam, Vinod Gopalan, Alfred King-Yin Lam. Oncogenic role of GAEC1 and its potential modulation with p53 in pathogenesis of colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5764. doi:10.1158/1538-7445.AM2017-5764
Abstract 5772: Developing a patient derived 3D co-culture model of prostate organoids to examine biological mechanisms involved in prostate cancer disparities
Authors: Marsili Joseph , Richards Zachary , Garcia Jason , Voisine Cindy , Nonn Larisa
Published: July 2017

The prostate is composed of a fibro-muscular stromal and glandular epithelial cells. Cross-talk between these cell populations is essential for normal prostate development and is involved in carcinogenesis. In the past year, an estimated 220,800 new cases of prostate cancer (PCa) were diagnosed and 27,540 died of PCa. PCa in African American men occurs more frequently, develops at a much earlier age, and is more aggressive and lethal compared to men of European descent. Both biological and socioeconomic factors likely contribute to this disparity. There is currently a paucity of relevant preclinical models in which to examine biological mechanisms in African American PCa. Patient-derived organoids are a recently developed in vitro preclinical model which facilitate examination of underlying mechanisms of disease etiology. However, current models of epithelial organoid growth lack the prostate stroma. To address this unmet need, we began development of a 3D organoid model that incorporated both primary human prostatic epithelial (PrE) and stromal (PrS) cells to better mimic the microenvironment of the organ. Prostate organoids were derived from cells collected from radical prostatectomy tissues from patients (UIC IRBs: 2006-0679 (PI Behm), (2013-0341(PI Nonn). Mixtures of single PrE-derived and single PrS-derived cells were seeded in 3D in matrigel. Bright-field microscopy of live organoids and histological examination of fixed organoids showed that the addition of PrS cells to the culture altered the shape of the organoids compared to the spherical phenotype observed in PrE only cultures. Immunofluorescent analysis of the basal cell marker p63 showed increased epithelial differentiation in the co-culture organoids. In summary, these data suggest that inclusion of prostate stroma in co-culture organoid models influences prostatic epithelial organoid development in vitro. Ongoing studies are focused on molecular analysis or organoid phenotypes and comparison of AA and EA-derived organoids.

Citation Format: Joseph Marsili, Zachary Richards, Jason Garcia, Cindy Voisine, Larisa Nonn. Developing a patient derived 3D co-culture model of prostate organoids to examine biological mechanisms involved in prostate cancer disparities [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5772. doi:10.1158/1538-7445.AM2017-5772
Abstract 5777: Translation of HPV mediated immortalization to cancer precision medicine
Authors: Dakic Aleksandra , Palechor-Ceron Nancy , Krawczyk Ewa , Yuan Hang , Suprynowicz Frank , Agarwal Seema , Schlegel Richard , Simic Vera , Sripadhan Praathibha , Chen Chen , Lu Jie , Hou Tung-Wei , Choudhury Sujata , Liu Xuefeng
Published: July 2017

The E6/E7 oncogenes of the high-risk HPVs are both necessary and sufficient to immortalize HFKs and their presence and expression is required for the continued proliferation of HPV-positive cervical cancer cells. We and others have shown previously that hTERT induced by E6 and cytoskeleton alteration by E7 are critical. Both E6 and feeder cells activate telomerase, while both E7 and Rock inhibitor (Y-27632) disrupt the actin cytoskeleton and inactivate Rho. Feeders and Y-27632 to induce unlimited cell proliferation of human keratinocytes. Unexpectedly, we observed that feeders and Y-27632 could be used to establish both normal and tumor cell cultures from non-keratinocyte tissues. This culture has been termed as “conditional reprogramming”, since CR cultures stop proliferating or terminally differentiate after its removal, depending on culture conditions. The Combination of CR and Organoids ( (Matrigel, air-liquid interface (ALI)) cultures represents next generation human cancer models and functional diagnostics for cancer precision medicine as described in August 2015 in the NCI precision medicine initiative and three nature review articles (Nat Rev Cancer. 2015 Dec; Nat Rev Genet. 2015 Jul; Nat Rev Clin Oncol. 2014 Nov.). The technique is relatively simple and has been reproduced in more than 50 laboratories (including an original and independent article in Science from Massachusetts General Hospital) (Science. 2014, 346(6216):1480-6.). Importantly, the CR technology can generate 2x10<6cells in a week from small biopsies, and can generate cultures from cryopreserved tissue and small biopsies. We therefore initiated studies to examine whether CR cultures reflected the biology and genotype of the original tumor and whether cultures might be used to predict clinical responses. Moreover, the epithelial cells can be propagated indefinitely in vitro, yet retain the capacity to become fully differentiated when placed into conditions that mimic their natural environment. Thus, the CR method significantly advances applications for regenerative medicine.

Citation Format: Aleksandra Dakic, Nancy Palechor-Ceron, Ewa Krawczyk, Hang Yuan, Frank Suprynowicz, Seema Agarwal, Richard Schlegel, Vera Simic, Praathibha Sripadhan, Chen Chen, Jie Lu, Tung-Wei Hou, Sujata Choudhury, Xuefeng Liu. Translation of HPV mediated immortalization to cancer precision medicine [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5777. doi:10.1158/1538-7445.AM2017-5777
Abstract 5796: Heparan sulfate proteoglycans mediate tumor cell invasion and metastasis
Authors: Qazi Henry , Shi Zhong-Dong , Song Jonathan W. , Cancel Limary M. , Munn Lance L. , Tarbell John M.
Published: July 2017

The surface proteoglycan/glycoprotein layer (glycocalyx) on tumor cells has been associated with cellular functions that can potentially enable invasion and metastasis. In addition, aggressive tumor cells with high metastatic potential have enhanced invasion rates in response to interstitial flow stimuli in vitro. Our previous studies suggest that heparan sulfate (HS) in the glycocalyx plays an important role in this flow mediated mechanostransduction and upregulation of invasive and metastatic potential. In this study, highly metastatic renal cell carcinoma cells were genetically modified to suppress HS production by knocking down its synthetic enzyme NDST1. Using modified Boyden chamber and microfluidic assays, we show that flow-enhanced invasion is suppressed in HS deficient cells. To assess the ability of these cells to metastasize in vivo, parental or knockdown cells expressing fluorescence reporters were injected into kidney capsules in SCID mice. Histological analysis confirmed that there was a large reduction (95%) in metastasis to distant organs by tumors formed from the NDST1 knockdown cells compared to control cells with intact HS. The ability of these cells to invade surrounding tissue was also impaired. The substantial inhibition of metastasis and invasion upon reduction of HS suggests an active role for the tumor cell glycocalyx in tumor progression.

Citation Format: Henry Qazi, Zhong-Dong Shi, Jonathan W. Song, Limary M. Cancel, Lance L. Munn, John M. Tarbell. Heparan sulfate proteoglycans mediate tumor cell invasion and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5796. doi:10.1158/1538-7445.AM2017-5796
Abstract 580: Nivolumab treatment of metastatic renal cancer patients impairs Tregs and potentiates NK function: The role of CXCR4 inhibition (“REVOLUTION Trial”)
Authors: Santagata Sara , Trotta Anna Maria , Napolitano Maria , Portella Luigi , Rossetti Sabrina , Perdona Sisto , Pignata Sandro , Scala Stefania
Published: July 2017

Despite encouraging results, Nivolumab response is not as wide as expected in renal cancer ( RCC). Among mechanisms of immunoresistance T-regulatory cells(Tregs) activity plays a central role. We previously showed that tumoral Tregs in RCC patients are more suppressive than healthy donors Tregs. Since tumoral Tregs express high level of CXCR4, the receptor antagonism was evaluated on patients Tregs revealing that CXCR4 antagonism reverted Treg suppressive activity( Santagata et al, submitted manuscript). To identify biomarkers informative and predictive of nivolumab efficacy Treg function, tumoral access and NK interactions was determined in nivolumab treated mRCC patients (“ Revolution” trial). At today eleven patients were analyzed for Tregs and NKs function at time 0-2-4 weeks-3-6 months of treatment. 2 patients died for unrelated causes, 5 Patients showed PR, 4 SD and 1 PD at six months. At time 0 Tregs function, evaluated as inhibition of T-effector (Teff) proliferation, varied among patients; two patients showed anergy reversed by Nivolumab treatment. Overall, increase in Teff proliferation, compatible with a decreased Tregs activity, was revealed during nivolumab treatment and, while the total peripheral Tregs was unaffected, a decrease in CD4+CD25+127lowFOXp3 highCD45RA - (suppressive Tregs) was detected. NK activity, evaluated as CD107a exposure, increased with nivolumab treatment with a concomitant reduction in the inhibitory receptors CD158a, PD-1 and CXCR4. Moreover patients Tregs and NKs were evaluated in vitro for the efficacy of CXCR4 inhibition in affecting cell function. CXCR4 antagonists suppressed the Treg mediated inhibition of Teff proliferation. In conclusion the evaluation of 11 patients enrolled in the REVOLUTION study revealed detectable variations on Tregs and NKs function relatively to Nivolumab treatment to be correlated with clinical outcome. Tregs suppressive function was impaired by inhibition of CXCR4 receptor.

Citation Format: Sara Santagata, Anna Maria Trotta, Maria Napolitano, Luigi Portella, Sabrina Rossetti, Sisto Perdona, Sandro Pignata, Stefania Scala. Nivolumab treatment of metastatic renal cancer patients impairs Tregs and potentiates NK function: The role of CXCR4 inhibition (“REVOLUTION Trial”) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 580. doi:10.1158/1538-7445.AM2017-580
Abstract 5807: Disseminated tumor cell clearance by the immune system
Authors: Piranlioglu Raziye , Lee Eun Mi , Ouzunova Maria , Arbab Ali S. , Rodriguez Paulo C. , Iskander Asm L. , Korkaya Hasan
Published: July 2017

The classical model of metastasis suggests that the tumor cell dissemination occurs late in tumor development, however accumulating evidence coming from mouse studies and clinical data provide striking evidence that tumor cells start to disseminate during the initial steps of tumor development. However, the dissemination from the primary area does not always result in metastasis. Due to the non-permissive nature of microenvironment in distant sites, these early disseminated tumor cells might be cleared or maintained in a non-proliferative/dormant state. The mechanism by which some early disseminated tumor cells colonize and generate metastatic growth while some remain dormant is not well known. In order to understand the underlying factors that may contribute to the metastatic growth, we performed time course experiments by utilizing murine mammary tumors (4T1 as metastatic and EMT6 as less metastatic) in a syngeneic mouse model. Luciferase expressing 4T1 or EMT6 tumor cells were orthotopically implanted into the fat pads and tumor cell dissemination was analyzed over 3-week time points. We determined that both 4T1 and EMT tumors disseminated as early as one to two-week post implantation, however only 4T1 tumor develop metastasis in distant organs. Moreover, we also resected primary tumors 1, 2 and 3-week post implantation of EMT6-Luci or 4T1-Luci tumors and observed distant metastasis via optical imaging of luciferase expression in live animals. Although the majority of 4T1 tumor-bearing mice (>80%) develop pulmonary metastasis when 4T1 tumors resected 2 and 3 weeks post-implantation only 10% of mice develop metastasis when primary tumor resected one-week post implantation. In contrast, EMT6 tumors following resection only relapsed in the primary tumor site but failed to develop metastasis. Furthermore, EMT6 tumor-bearing mice efficiently cleared tail vein injected EMT6-luci cells in the lungs. We investigated the possible mechanism by which EMT6 tumor-bearing mice clears disseminated tumor cells in the lung. We provide evidence that pulmonary infiltrated mMDSCs mediate tumor cell killing via secretion of high levels of cytotoxic granules, granzyme A, granzyme B, perforin. This was confirmed by mouse transcriptome and qPCR analyses as well as biochemistry using in vivo samples and in vitro co-culture samples. Our studies provide a new paradigm in the understanding of the fate of disseminated tumor cells in secondary organs and the role of the immune system in this process.

Citation Format: Raziye Piranlioglu, Eun Mi Lee, Maria Ouzunova, Ali S. Arbab, Paulo C. Rodriguez, Asm L. Iskander, Hasan Korkaya. Disseminated tumor cell clearance by the immune system [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5807. doi:10.1158/1538-7445.AM2017-5807
Abstract 5810: microRNAs targeting EMT transcription factors may be associated to lymph node metastasis in breast cancer tumors
Authors: Pérez-Moreno Elisa , Valarezo Gabriela , Zavala Valentina , Fernández Wanda , Carvallo Pilar
Published: July 2017

Breast cancer is the leading cause of cancer-associated deaths in women. Lymph node near to the primary breast tumor have a high chance of developing a secondary tumor, representing one of the first signs of metastasis in breast cancer. Metastasis is promoted by epithelial-mesenchymal transition (EMT), process leaded by the transcription factors SNAIL, SLUG, ZEB and TWIST. MicroRNAs are small non-coding RNAs, whose expression has been demonstrated to be altered in different cancer types. Because of their ability to regulate large sets of genes involved in cancer growth and metastasis, microRNAs have emerged as candidate molecular biomarkers and novel therapeutic targets. The aim of this study is to identify microRNAs differentially expressed in breast tumors in relation to EMT-transcription factor expression and lymph node metastasis, and that are involved in epithelial-mesenchymal transition. For this purpose, we used microRNA microarray data from 50 fresh frozen breast tumors with different tumor grades (1 to 3), 28 from patients with lymph node metastasis. Microarray data analysis using RankProd (R package) revealed approximately 40 microRNAS down-regulated in breast tumors with expression of EMT-transcription factors (p<0.05). Several of the identified microRNAs have been previously described to regulate early steps of metastasis in different cancer types, like members of the miR-200 family (miR-200c and miR-141), miR-205 and miR-30a. In addition we found other microRNAs not previously associated to metastasis. Our results suggest that down regulation of specific microRNAs, and up regulation of their targets, may induce a metastatic behavior of tumor cells of the primary breast tumors, promoting invasion and colonization of the lymph nodes. In this sense, the changes in the expression of microRNAs may serve as biomarkers of breast cancer prognosis in patients.

Citation Format: Elisa Pérez-Moreno, Gabriela Valarezo, Valentina Zavala, Wanda Fernández, Pilar Carvallo. microRNAs targeting EMT transcription factors may be associated to lymph node metastasis in breast cancer tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5810. doi:10.1158/1538-7445.AM2017-5810
Abstract 5848: Fucoidan adminstration attenuates radiation pneumonitis and fibrosis through reducing inflammatory cytokine expression in lung tissue
Authors: Wu Szu-Yuan
Published: July 2017

Purpose: There is no well-known compound to prevent radiation pneumonitis (RP) or prophylactically use to attenuate the toxicity of RP. RP is one of major complications in chest part irradiation and limitations of radiation dose. Fucoidan is a sulfated polysaccharide found mainly in various species of brown algae and brown seaweed. Recent studies have demonstrated its anti-inflammation effect. Since RP is an radiation induced inflammatory and fibrosis related complication, we investigated the effect of fucoidan adminstration on radiation pneumonitis and fibrosis.

Materials and Methods: In this study, we compared the pneumonitis and fibrosis in lung tissue specimen between the lungs irradiated (10Gy/one shot) C57BL/6 mice with or without Fucoidan adminstration (200mg/kg /day, oral gavage for 14 days).

Results: The results demonstrated that fucoidan adminstration attenuates pneumonitis and fibrosis in lung tissue. We found the accumulation of neutrophils in irradiated lung tissue was decreased, and the radiation induced Masson trichrome staining fibrosis was decreased. We also investigate the inflammatory -cytokine profiles in the irradiated lung tissue by protein array. The results revealed that fucoidan adminstration changes the inflammatory -cytokine expression pattern in in the irradiated lung tissue.

Conclusions: This study demonstrated that inflammatory -cytokine expression after whole lung irradiation is correlated with neutrophil accumulation and radiation induced fibrosis. Interestingly, Fucoidan adminstration changes the inflammatory -cytokine expression pattern which may lead to attenuates radiation pneumonitis and fibrosis. It is potential to be a therapeutic molecular to attenuate or prevent radiation pneumonitis.

Note: This abstract was not presented at the meeting.

Citation Format: Szu-Yuan Wu. Fucoidan adminstration attenuates radiation pneumonitis and fibrosis through reducing inflammatory cytokine expression in lung tissue [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5848. doi:10.1158/1538-7445.AM2017-5848
Abstract 5856: Molecular mechanism underlying breast cancer cell radioresistance: role of radiation-induced NF-KB self-sustenance
Authors: Yu Hui , Aravindan Natarajan , Xu Ji , Natarajan Mohan
Published: July 2017

Understanding the mechanism of breast cancer cell radioresistance and the associated selection pressure, tumor recurrence, would help us to develop molecular interventions and achieve disease/progression-free survival in patients with cancer. In the present study, we investigated the molecular mechanisms prompted in breast cancer cells in response to clinical radiation and the associated coordination of intra- and inter-cellular signaling that orchestrate radioresistance and tumor relapse/recurrence. Our findings showed that clinical doses of radiation increase NF-κB transcription and DNA-binding activity. Further, radiation exposure resulted in de novo TNF-α synthesis and secretion. An NF-κB gene manipulation approach revealed that NF-κB triggered NF-κB-mediated TNF-α transcriptional activity. NF-κB DNA binding kinetic analysis demonstrated both sustained and dual phase NF-κB activation with radiation. Radiation prompted NF-κB-dependent synthesis/secretion of TNF-α, a requisite for the second phase of NF-κB activation and maintenance. Radiation-associated NF-κB-dependent secretion of TNF-α from irradiated cells also activated NF-κB in the non-irradiated, non-targeted bystander cells. Together, these findings demonstrated that radiation-triggered NF-κB-dependent TNFα secretion is critical for self-sustenance of NF-κB (through autocrine positive feedback signaling) and for coordinating bystander response (through intercellular signaling: de novo activation of NF-κB in bystander cells) after radiation exposure. Further, the data suggest that this NF-κB→TNF-α→NF-κB feedback-dependent self-sustenance of NF-κB could drive the selection pressure (proliferation, clonal expansion) of surviving breast cancer cells and subsequent tumor relapse/recurrence.

Note: This abstract was not presented at the meeting.

Citation Format: Hui Yu, Natarajan Aravindan, Ji Xu, Mohan Natarajan. Molecular mechanism underlying breast cancer cell radioresistance: role of radiation-induced NF-KB self-sustenance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5856. doi:10.1158/1538-7445.AM2017-5856
Abstract 5876: Osimertinib (AZD9291), a mutant-selective EGFR inhibitor, reverses ABCB1-mediated drug resistance in cancer cells
Authors: Zhang Xiao-Yu , Lei Zi-Ning , Zhang Yun-Kai , Wang Yi-Jun , Gupta Pranav , Zeng Leli , Xu Megan , Wang Xiu-Qi , Yang Dong-Hua , Chen Zhe-Sheng
Published: July 2017

In recent years, tyrosine kinase inhibitors (TKIs) have been shown capable of inhibiting the ATP-binding cassette (ABC) transporter-mediated multidrug resistance (MDR). In this study, we determine whether osimertinib, a novel selective, irreversible EGFR (epidermal growth factor receptor) TKI inhibitor, could reverse ABC transporter-mediated MDR. The results showed that at non-toxic concentrations, osimertinib significantly sensitized both ABCB1-transfected and drug-selected cell lines to substrate anticancer drugs colchicine, paclitaxel, and vincristine. Osimertinib significantly increased the accumulation of [3H]-paclitaxel in ABCB1 overexpressing cells by blocking the efflux function of ABCB1 transporter. In contrast, no significant alteration in the expression levels and localization pattern of ABCB1 was observed when ABCB1 overexpressing cells were exposed to 0.3μM osimertinib for 72 h. In addition, ATPase assay showed osimertinib stimulated ABCB1 ATPase activity. Molecular docking and molecular dynamic simulations showed osimertinib has strong and stable interactions at transmembrane domain of human homology ABCB1. Taken together, our findings suggest that osimertinib, a clinically approved third-generation EGFR TKI, can reverse ABCB1-mediated MDR, which supports the combination therapy with osimertinib and ABCB1 substrates may potentially be a novel therapeutic treatment in ABCB1-positive drug resistant cancers.

Citation Format: Xiao-Yu Zhang, Zi-Ning Lei, Yun-Kai Zhang, Yi-Jun Wang, Pranav Gupta, Leli Zeng, Megan Xu, Xiu-Qi Wang, Dong-Hua Yang, Zhe-Sheng Chen. Osimertinib (AZD9291), a mutant-selective EGFR inhibitor, reverses ABCB1-mediated drug resistance in cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5876. doi:10.1158/1538-7445.AM2017-5876
Abstract 5910: Influence of 3D- cultured prostate cancer and stromal cells on drug response
Authors: Gales Dominique N. , Miller Sonni , Samuel Temesgen , Yates Clatyon
Published: July 2017

Background: Response to chemotherapeutic drugs presents a significant challenge to effective treatment of advanced prostate cancer. Exploration of therapeutic response has focused largely on the tumor cell. However, evidence suggests mechanisms that involve the tumor microenvironment mediate response to therapeutics. Therefore, it is imperative to mimic the in vivo tumor microenvironment, to bridge the gap between in vitro studies and the development of therapeutics. The concept of three-dimensional (3D) cell culture and co-culture of multiple cell types is critical to predict therapeutic efficacy and outcome. Thus, these studies sought to determine the distinct responses to anti-cancer drugs in 3D cell spheroids.

Methods: In this study, we utilized metastatic prostate cancer cells (PC3) and fluorescently labeled prostate bone stromal cell line (HS27a) to generate tumor spheroids. Tumor spheroid viability was measured at 72 and 120 h using the CellTiter-Glo 3D assay after treatment with three common anti-cancer drugs, Docetaxel (4ηM), Enzalutamide (30μM), and Bicalutamide (30μM).Treatments were compared with spheroids originated from HS27a and PC3 cells alone and in co-culture. The cell viability percentage was normalized using the untreated cells as 100% cell survival.

Results: Spheroid formation increased resistance to Docetaxel, Enzalutamide, and Bicalutamide. However, PC3 spheroid cells was significantly higher compared to HS27a and co-culture spheroids. For all three spheroids, as the size of the spheroids increase, cellular resistance to anti-cancer and hormone drugs increased.

Conclusion: The findings in this study demonstrated that the use for 3D-cell cultures influence many aspects of PCa behavior including, proliferation rates, and response to chemotherapeutics.

Note: This abstract was not presented at the meeting.

Citation Format: Dominique N. Gales, Sonni Miller, Temesgen Samuel, Clatyon Yates. Influence of 3D- cultured prostate cancer and stromal cells on drug response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5910. doi:10.1158/1538-7445.AM2017-5910
Abstract 5911: Omentin drives metabolic shift in ovarian cancer cells in the omental tumor microenvironment
Authors: Yeung Chi Lam Au , Achreja Abhinav , Zhao Hongyun , Yeung Tsz-Lun , Schmandt Rosemarie , Yip Daniel K. , Lu Karen H. , Nagrath Deepak , Mok Samuel C.
Published: July 2017

Advanced stage high grade serous ovarian cancer (HGSC) metastasizes preferentially to the omentum, which is a well-vascularized fold of peritoneal tissue and is a major site of adipose tissue accumulation. The mechanisms by which omental adipose tissue interact with ovarian cancer cells and promotes tumor growth and disease progression are not entirely clear. We previously showed that a novel adipokine called omentin (Intestinal Lactoferrin Receptor ITLN1) produced by the omental adipose tissue was significantly down-regulated in patients with HGSC compared with those in BMI matched healthy individuals, and omentin suppressed ovarian cancer cell growth only in the presence of adipocytes in vitro. Since we and others demonstrated that omentin induced insulin-dependent glucose uptake exclusively in adipocytes, we therefore hypothesized that omentin may suppress ovarian cancer growth via driving metabolic shift in ovarian cancer cells in the omental microenvironment. Using a cancer cell/adipocyte-co-culture model, we demonstrated that omentin reduced both glucose uptake and lactate secretion in ovarian cancer cells when they were co-cultured with adipocytes but not with other stromal cell types, suggesting that omentin-induced glucose uptake in adipocytes may deplete the surrounding glucose that fuels the glucose-addicted ovarian cancer cells in the omental microenvironment and thus drive metabolic shift in ovarian cancer cells. To delineate the underlying mechanism by which omentin induced glucose uptake in adipocytes, expression levels of the adipocyte-specific glucose transporter GLUT4 induced by omentin in adipocytes and the effect of GLUT4 silencing in adipocytes using GLUT4 specific siRNAs on ovarian cancer growth were examined. The results showed that omentin up-regulated GLUT4 in adipocytes and GLUT4 silencing in adipocytes abrogated the effects of omentin on glucose uptake in adipocytes and ovarian cancer cell growth in the co-culture model. Taken together, this study shows that omentin plays an important role in driving metabolic shift in ovarian cancer cells in the omental microenvironment. Therapeutic strategies based on up-regulating omentin in ovarian cancer patients may inhibit ovarian cancer progression and improve patient survival rates.

Note: This abstract was not presented at the meeting.

Citation Format: Chi Lam Au Yeung, Abhinav Achreja, Hongyun Zhao, Tsz-Lun Yeung, Rosemarie Schmandt, Daniel K. Yip, Karen H. Lu, Deepak Nagrath, Samuel C. Mok. Omentin drives metabolic shift in ovarian cancer cells in the omental tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5911. doi:10.1158/1538-7445.AM2017-5911
Abstract 5942: Collagen proly-hydroxylase regulates breast cancer progression and chemoresistance
Authors: Xiong Gaofeng , Xu Ren
Published: July 2017

Collagen prolyl hydroxylation (P4H), an enzyme hydroxylating proline residues in -X-Pro-Gly- sequences, is a potential therapeutic target for the disorders associated with increased collagen deposition. Upregulation of collagen prolyl-4-hydroxylase in cancer cells is required for breast cancer promotion. P4HA1 mRNA levels were significantly upregulated in triple-negative breast cancer (TNBC) compared to other cancer subtypes. Increased mRNA levels of P4HA1 correlated with poor clinical outcome in breast cancer patients. Silencing P4HA1 expression or treatment with the P4HA inhibitor significantly inhibited cell proliferation and suppressed aggressive phenotypes of breast cancer cells in 3D culture, accompanied by reduced deposition of collagen. We found that activation of the P4HA1 in TNBC cells enhanced cancer cell stemness. We also found that knockdown of P4HA1 inhibited mammary tumor growth and metastasis to lungs in xenograft models. Inhibition of P4HA1 activity with the P4H inhibitor sensitized TNBC to the docetaxel treatment in the patient-derived organoid (PDO) and xenograft models. Therefore, these results suggest the crucial role of P4HA1 in breast cancer progression and that targeting collagen P4H is a promising strategy to inhibit tumor progression and to sensitize TNBC to chemotherapy.

Citation Format: Gaofeng Xiong, Ren Xu. Collagen proly-hydroxylase regulates breast cancer progression and chemoresistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5942. doi:10.1158/1538-7445.AM2017-5942
Abstract 5946: Universal consent for biospecimens: A novel electronic/video consent
Authors: Naeim Arash , Wenger Neil , Petruse Antonia , Sanchez Liliana , Sharif Azita , Dry Sarah
Published: July 2017

Background: Developing innovative, efficient and institutionally scalable bio-specimen consent for remnant tissue that meets the NIH consent guidelines for genomic and molecular analysis is essential for precision medicine efforts in cancer. Solutions in this arena need to satisfy the needs of patients, researchers, ethicists, IRB and compliance leadership, while fitting as seamlessly as possible into existing clinical workflows.

Methods: UCLA developed a video-application kiosk-based approach for providing universal consent to repurpose clinical remnant bio-specimen for research. The process was designed to be self-service, comprehensive yet fast (mean shorter than 5 minute for completion). The consent additionally asked the patient if they were willing to be contacted directly for future research projects. This approach was piloted with 474 patients who were coming in for routine services in laboratory medicine, radiology, oncology, and hospital admissions. Of the pilot population, 175 individuals had targeted surveys to evaluate drivers for opting-in or opting-out of the consent for allowing the collection and use of their remnant tissue for research. The cognitive survey was online and presented immediately after the consent process was completed.

Results: The opt-in rate for the pilot was 90.7%, and 56% agreed to direct contact for future research. Only 7% needed help navigating the online consent process. Of the subgroup of pilot population who completed the targeted survey, there was no difference between individuals who opted in and out regarding ease of use, of the consent application with about 75% stating it provided mostly or very useful information, 90% stating it was mostly or very easy to understand, and 85% stating they trusted the information. However, there were significant differences between those that opted-in and opted-out in their beliefs concerning usefulness of tissue, trusting researchers, importance of contributing to science and privacy risk with those opting in strongly supporting these beliefs (>90%) compared to those that opted out (<40%), p<0.001.

Conclusions: Video-application approach for allowing individuals to consent for remnant specimens to be collected and used for research, including cancer research, can be efficient, patient-centric and meet the NIH requirements. This method could increase the availability of blood and tissue for cancer research and should be tested for scalability as an enterprise solution.

Citation Format: Arash Naeim, Neil Wenger, Antonia Petruse, Liliana Sanchez, Azita Sharif, Sarah Dry. Universal consent for biospecimens: A novel electronic/video consent [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5946. doi:10.1158/1538-7445.AM2017-5946