bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 881: CD44-specific supramolecular hydrogels for fluorescence molecular imaging of EMT induced BRAF  mutant thyroid cancer cells
Authors: Byeon Hyung Kwon , Ku Minhee , Yang Yeon Ju , Cho Min Hee , Oh Yoojung , Kim Jae Wook , Ban Myung Jin , Kim Ji-Hoon , Kim Da Hee , Kim Joo Hyun , Yang Jaemoon , Koh Yoon Woo
Published: July 2017

Previously, the authors have identified that the acquired drug resistance to BRAF inhibitor, PLX4032 in BRAF (V600E) mutant anaplastic thyroid cancer promotes not only tumor progression and proliferation, but also migration and invasion of cancer through upregulated epithelial-to-mesenchymal transition (EMT). The underlying mechanism to the acquired resistance to BRAF inhibition involves c-Met-mediated reactivation of PI3K/AKT pathway. Therefore combinatorial dual targeted therapy of BRAF and c-Met inhibition has shown to reverse EMT and show maximal antitumor effect. Previously, the authors have developed a novel in vivo imaging strategy using CD44-targetable near-infrared (NIR)-sensitive supramolecular hydrogels (NIRSHs) for the recognition of CD44-expressing cancer cells. In the present study, we applied this NIR-sensitive molecular imaging probe in detecting the upregulated EMT changes in PLX4032-treated 8505C cells. The CD44-targetable NIRSHs were fabricated by polyplexing Cy5.5-conjugated polyethyleimine and hyaluronic acid in an aqueous medium. Ectopic xenograft mouse models were prepared by injecting 8505C cells at the flank of male athymic nude BALB/c mice, aged 6 weeks. After confirming tumor formation at 3 weeks post-injection, the mice were randomly divided into four groups and were each treated under different conditions; DMSO, PLX4032, PHA665752, PLX4032 and PHA665752. After 3 weeks, the pre-established NIRSH probes were injected and confirmed by IVIS imaging. The injected NIRSH probes showed highest uptake in the PLX4032 single treatment group and lowest uptake in the PLX4032 and PHA665752 combination group. Sizes of tumor were verified by MRI which showed correlations with the NIRSH fluorescence imagings. The results suggest that CD44-targetable NIRSHs imaging shows potential as a non-invasive in vivo imaging tool in detecting the increased invasion potential of cancer cells and monitoring appropriate therapeutic effects.

Citation Format: Hyung Kwon Byeon, Minhee Ku, Yeon Ju Yang, Min Hee Cho, Yoojung Oh, Jae Wook Kim, Myung Jin Ban, Ji-Hoon Kim, Da Hee Kim, Joo Hyun Kim, Jaemoon Yang, Yoon Woo Koh. CD44-specific supramolecular hydrogels for fluorescence molecular imaging of EMT induced BRAF  mutant thyroid 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 881. doi:10.1158/1538-7445.AM2017-881
Abstract 887: Magnetic relaxometry detection of stealth, antibody-targeted micellar iron oxide nanoparticles in-vivo
Authors: Aburto Rebeca Romero , Sokolov Konstantin , Kulp Adam M. , Vreeland Erika C. , Lu Zhen , Bast Robert C. , Hazle John D. , Mathieu Kelsey B.
Published: July 2017

Magnetic relaxometry (MRX) has the potential to provide unprecedented sensitivity in early detection of cancer by sensing changes in magnetic relaxation of iron oxide (Fe3O4) nanoparticles targeted to cancer biomarkers and is expected to exceed the detection limits of established clinical modalities. MRX uses superconducting quantum interference device (SQUID) sensors to measure Neél relaxation of bound particles. Our strategy was to develop molecular specific Fe3O4 nanoparticles using amphiphilic functionalized phospholipids that allow for clinical translation of the MRX technology. To accomplish this, we used automated, controlled rate, direct infusion of an organic phase mixture of phospholipids and nanoparticles into water to produce monodisperse micellar nanoparticles with a mean diameter of 75±12 nm and surface charge of -10mV. The particles were determined to be stable in various biological media, including human plasma, for more than 24 hours with no detectable formation of a protein corona. Furthermore, in-vivo studies in healthy mice showed blood circulation times of more than 2 hours, as well as minimal MRX signals during this time. Additionally, we developed maleimide conjugation chemistry for epidermal growth factor receptor (EGFR) antibody attachment to micellar nanoparticles. We have achieved molecular specific labeling of cancer cells over-expressing EGFR. In the future, we will evaluate the MRX signal impact from injecting EGFR-conjugated nanoparticles into tumor-bearing mice.

Citation Format: Rebeca Romero Aburto, Konstantin Sokolov, Adam M. Kulp, Erika C. Vreeland, Zhen Lu, Robert C. Bast, John D. Hazle, Kelsey B. Mathieu. Magnetic relaxometry detection of stealth, antibody-targeted micellar iron oxide nanoparticles in-vivo [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 887. doi:10.1158/1538-7445.AM2017-887
Abstract 895: Fucosylation inhibits invadopodia formation and melanoma invasion
Authors: Keeley Tyler S. , Lau Eric , Yang Shengyu
Published: July 2017

Background: Melanoma is the most deadly skin cancer and metastasis is essentially responsible for all melanoma related death. Recently, fucose treatment has been shown to inhibit melanoma cell migration and invasion in vitro, and melanoma metastasis in a mouse model, suggesting that protein fucosylation could be exploited to prevent or suppress melanoma metastasis. Invadopodia are proteolytic actin-rich protrusions used by metastatic melanoma cells to degrade extracellular matrix and to promote invasion and metastasis. In this study, we investigated the mechanisms by which fucosylation regulate melanoma invasion and invadopodia formation

Method: The melanoma cells were treated with L-fucose-supplemented medium or infected with fucokinase-expressing lentivirus to increase protein fucosylation. shRNAs were also employed to knockdown fucokinase and to inhibit fucosylation in melanoma cells. The effects of fucosylation on invadopodia formation and extracellular matrix degradation were determined by phalloidin staining and degradation of fluorescently labeled gelatin thin film.

Results: Here, we show that the addition of L-fucose to complete media inhibits the ability of melanoma cells to form invadopodia in a dose-dependent manner. The overexpression of hFUK in WM793 cells showed a similar result as L-fucose treatment, where invadopodia formation was significantly reduced. In addition, we observed a significant decrease in the percentage of cells that have invadopodia during L-fucose supplementation or hFUK overexpression. In an invadopodia precursor assay, the hFUK-overexpressing cell line exhibited delayed FBS-stimulated invadopodia formation.

Conclusions: Our data suggest that fucose or the overexpression of hFUK reduces melanoma invasion by inhibiting invadopodia formation. Given that invadopodia formation was hindered within an hour after stimulation, we believe that invadopodia initiation is affected by increased fucosylation, resulting in fewer invadopodia and slower extracellular matrix degradation.

Citation Format: Tyler S. Keeley, Eric Lau, Shengyu Yang. Fucosylation inhibits invadopodia formation and melanoma invasion [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 895. doi:10.1158/1538-7445.AM2017-895
Abstract 9: Suppression of the invasion and growth of human head and neck squamous cell carcinomas via regulating STAT3 signaling and miR-21/β-catenin axis with HJC0152
Authors: Wang Yu , Wang Sinan , Wu Yansheng , Zhou Jia , Shen Qiang , Zhou Xuan
Published: July 2017

Signal transducer and activator of transcription 3 (STAT3) is involved in the tumor growth and metastasis of human head and neck squamous cell carcinoma (HNSCC) and is therefore a target with therapeutic potential. In this study, we show that HJC0152, a recently developed anticancer agent and a STAT3 signaling inhibitor, exhibits promising antitumor effects against HNSCC both in vitro and in vivo via inactivating STAT3 and downstream microRNA-21/β-catenin axis. HJC0152 treatment efficiently suppressed HNSCC cell proliferation, arrested the cell cycle at the G0/G1 phase, induced apoptosis, and reduced cell invasion in both SCC25 and CAL27 cell lines. Moreover, HJC0152 inhibited nuclear translocation of phosphorylated STAT3 at Tyr705 and decreased VHL/β-catenin signaling activity via regulation of microRNA-21. Loss-of-function of VHL remarkably compromised the antitumor effect of HJC0152 in both cell lines. In our SCC25-derived orthotopic mouse models, HJC0152 treatment significantly abrogated STAT3/β-catenin expression in vivo, which leading to a global decrease of tumor growth and invasion. With its favorable aqueous solubility and oral bioavailability, HJC0152 holds the potential to be translated into the clinic as a promising therapeutic strategy for patients with HNSCC.

Citation Format: Yu Wang, Sinan Wang, Yansheng Wu, Jia Zhou, Qiang Shen, Xuan Zhou. Suppression of the invasion and growth of human head and neck squamous cell carcinomas via regulating STAT3 signaling and miR-21/β-catenin axis with HJC0152 [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 9. doi:10.1158/1538-7445.AM2017-9
Abstract 901: The mechanobiome of pancreatic cancer: a viable, targetable drug space
Authors: Surcel Alexandra , Schiffhauer Eric S. , Thomas Dustin , Zhu Qingfeng , Anders Robert , Robinson Douglas
Published: July 2017

Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease with an annual mortality rate of 37,000 in the US alone. Its 5-year survival rate of ~6% remains relatively unchanged over the past 4 decades. Because PDAC has elevated rates of de novo and acquired resistance to traditional chemotherapies, we are exploring new drug spaces that target metastatic hepatic disease, the primary mortality factor for patients. Specifically, we have identified the cluster of proteins that sense and respond to mechanical stimuli - collectively known as the mechanobiome. This cytoskeletal machinery is responsible in large part for endowing metastatic cells with the ability to navigate through different tissue types. Based on their mechanoresponsiveness profile, we have predicted and identified via western blot and immunohistochemistry proteins in the mechanobiome that are upregulated in patient-derived pancreatic tissues. These proteins include non-muscle myosin IIA and IIC, alpha-actin-4, and filamin B. We are performing knockdown and overexpression studies of these isoforms on 2D and 3D behaviors - cell mechanics, migration, invasion, and tissue sphere formation and are poised to begin mouse studies on the metastatic sufficiency in altering the expression of these proteins. In addition, we are testing 4-HAP, a small molecule mechanics modulator that we previously identified that targets myosin IIC, thus affecting PDAC mechanics in PDAC murine models. Limited studies reveal that 4-HAP reduces metastasis. These data suggest that direct targeting of the mechanobiome may eventually expand treatment strategies for pancreatic cancer.

Citation Format: Alexandra Surcel, Eric S. Schiffhauer, Dustin Thomas, Qingfeng Zhu, Robert Anders, Douglas Robinson. The mechanobiome of pancreatic cancer: a viable, targetable drug space [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 901. doi:10.1158/1538-7445.AM2017-901
Abstract 903: CXCR4 and CXCR7 play distinct and overlapping roles in prostate cancer dissemination to bone
Authors: Roy Sounak , Valkenburg Kenneth C. , Pienta Kenneth J.
Published: July 2017

Approximately 28,000 men die of prostate cancer (PCa) each year in the US, and 90-100% of them will be due to bone metastasis. It has been demonstrated that PCa cells detach from the primary tumor and “home” to bone. Osteoblasts (bone-forming cells) secrete the chemoattractant stromal-derived factor 1 (SDF-1, also known as CXCL12), and PCa cells express the receptors for SDF-1, C-X-C chemokine receptors 4 and 7 (CXCR4 and CXCR7). PCa cells use these receptors to enter the chemoprotective bone microenvironment, specifically, the hematopoietic stem cell (HSC) niche in the endosteum. While small molecule inhibition of CXCR4 can mobilize PCa cells from the bone marrow microenvironment, it is unknown whether CXCR7 plays a compensatory role when CXCR4 is antagonized. SDF-1 is known to bind with greater affinity to CXCR7 compared to CXCR4 indicating a possible role of CXCR7 in tumor migration to the endosteal niche. Therefore, it is crucial to determine the individual and combined roles of CXCR4 and CXCR7 in the formation and treatment of PCa bone metastases. To answer these questions, first we assessed CXCR4 and CXCR7 expression across a panel of prostate cancer cell lines. We then transiently overexpressed and knocked down CXCR4 and CXCR7 in PCa cells and found CXCR4 and CXCR7 expression in vitro to directly correlate to cell viability and migration in PC3 PCa cells. To further test dissemination and metastasis in bone, we are making stable overexpression and knockout cells and will perform in vivo metastasis experiments. We have developed a novel immunofluorescence protocol for detecting and quantifying tumor cells in murine blood and bone marrow and will use this technique to determine the ability of PCa cells to disseminate to bone in the presence or absence of CXCR4 and/or CXCR7. This will be quickly translatable because we are currently running a first-in-prostate cancer clinical trial to determine whether small molecule inhibition of CXCR4 to mobilize PCa tumor cells from the bone marrow, combined with docetaxel, will benefit metastatic PCa patients. If we determine that CXCR7 plays a compensatory role in dissemination and/or metastasis, we will need to add CXCR7 inhibition to our treatment strategy to obtain the most efficient benefit for patients.

Citation Format: Sounak Roy, Kenneth C. Valkenburg, Kenneth J. Pienta. CXCR4 and CXCR7 play distinct and overlapping roles in prostate cancer dissemination to bone [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 903. doi:10.1158/1538-7445.AM2017-903
Abstract 906: PATZ1 promotes migration and invasion of thyroid cancer cells through upregulation of the activity of plasminogen activator and matrix metalloproteinases
Authors: Iesato Asumi , Nakamura Teruo , Uehara Tsuyoshi , Izumi Hiroto , Ito Ken-ichi
Published: July 2017

Background: PATZ1, a transcription regulator, has been reported to function as oncogene or tumor suppressor in several human malignant neoplasms. The aim of the study was to examine involvement of PATZ1 in carcinogenesis and progression of thyroid cancer.

Materials and methods: PATZ1 expression in 165 clinical thyroid specimens obtained from 87 patients were evaluated by IHC. We examined the function of PATZ1 using an immortalized normal thyroid follicular epithelial cell line (Nthy-ori 3-1), and four thyroid cancer cell lines; TPC-1 (papillary cancer), FTC-133 (follicular cancer), FRO and ACT-1 (anaplastic cancer). Knockdown of PATZ1 in Nthy-ori 3-1 and thyroid cancer cell lines was performed with siRNA-PATZ1, and overexpression of PATZ1 in thyroid cancer cell lines was performed with pcDNA3-FLAG-PATZ1.

Results: In the IHC analysis of clinical specimens, nuclear PATZ1 expression in anaplastic carcinoma was significantly less frequently than that in normal thyroid tissue or well-differentiated carcinoma (Table 1). Knockdown of PATZ1 in Nthy-ori 3-1 and differentiated thyroid cancer cell lines (TPC-1, FTC-133) induced morphological change of the cells and significant increase of cell proliferation and migration as well as moderate increase of invasion. Furthermore, the expression of u-PA, MMP2 and MMP11 was increased by inhibition of PATZ1 in TPC-1 and FTC-133. On the other hand, overexpression of PATZ1 in anaplastic cancer cell lines (FRO and ACT-1) decreased proliferation and migration of the cells along with the decreased expression of u-PA, MMP2, MMP9 and MMP11.

Conclusion: Our study suggests that PATZ1 might play an important role as a tumor suppressor in thyroid follicular epithelial cells and might be involved in progression of thyroid cancer.

Expression of nuclear PATZ1 in thyroid tumortypeNormal thyroid/ goiter (N=68)Follicular adenoma (N=5)Papillary cancer (N=39)Follicular cancer (N=8 )Poorly differentiated cancer (N=12)Anaplastic cancer (N=28)Medullary cancer (N=5)positive (%)68(100%)4(80.0%)35(89.7%)5(62.5%)7(58.3%)3(10.7%)4(80.0%)negative (%)0(0%)1(20.0%)4(10.3%)3(37.5%)5(41.7%)25(89.3%)1(20.0%)

Citation Format: Asumi Iesato, Teruo Nakamura, Tsuyoshi Uehara, Hiroto Izumi, Ken-ichi Ito. PATZ1 promotes migration and invasion of thyroid cancer cells through upregulation of the activity of plasminogen activator and matrix metalloproteinases [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 906. doi:10.1158/1538-7445.AM2017-906
Abstract 91: Molecular insight into drug resistance in acute myeloid leukemia: An in-silico and in-vitro approach
Authors: Desai Urja , Mirza Sheefa , Parikh Sonia , Rawal Rakesh
Published: July 2017

INTRODUCTION:

Drug resistance of leukemic stem cells is a surmountable obstacle to effective chemotherapy in acute myeloid leukemia (AML). P-glycoprotein (Pgp) and FLT3 undoubtedly contribute to worse prognosis and mechanisms of these proteins lead to shorter survival and chemotherapy resistance. Due to heterogeneity of stem cells in AML, outcome of patients with a normal karyotype is highly challenging and changes in drug resistance genes likely to be identified which will contribute to a better understanding of the disease biology, eventually leading to the development of alternative therapeutic approaches regarding drug resistance in AML.

METHODOLOGY:

Blood and Bone-marrow samples were collected from newly diagnosed adult AML patients with normal karyotype. Gene expression analysis of MDR genes like Pgp, MRP1, BCRP and LRP was done by qRT-PCR in FLT3-ITD+/- and CD34+/- subgroups. Higher expression of Pgp amongst all MDR genes instigated to undertake in-silico analysis of Pgp with natural compound library using YASARA. MTT assay was used to find out IC50 value of natural compounds shortlisted from in-silico analysis and used alone or with cytarabine on THP-1 cell line and in primary AML stem cells. Pgp inhibitory activity of natural compounds was assessed by fluorimetric MDR assay.

RESULTS:

A significant upregulation of Pgp expression was observed in FLT3-ITD+, CD34+ group after cytarabine exposure during their induction (7+3) therapy. Docking analysis revealed curcumin, hesperidin and silymarin having better binding affinity with Pgp as compared to its known inhibitor Verapamil. Furthermore, these selected compounds were used in-vitro, and results of MTT assay suggested 6.1µM, 40.73µM, 63.09µM and 54.95µM IC50 values of cytarabine, curcumin, hesperidin and silymarin respectively in THP-1 cell line. Combination index (CI) analysis revealed that curcumin (25µM), hesperidin (30µM) and silymarin (35µM) having synergistic effect with cytarabine and decreased IC50 value of cytarabine (1µM). In addition to this, Silymarin at 250nM and 500nM concentration showed equivalent inhibition of Pgp as compared to control Verapamil.

CONCLUSION:

A combination of in-silico and in-vitro screening revealed Curcumin, Hesperidin and Silibinin can be used as MDR modulators as well as chemosensitizer to reduce the cytotoxicity profile and drug resistance in AML.

Citation Format: Urja Desai, Sheefa Mirza, Sonia Parikh, Rakesh Rawal. Molecular insight into drug resistance in acute myeloid leukemia: An in-silico and in-vitro approach [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 91. doi:10.1158/1538-7445.AM2017-91
Abstract 911: Destabilizing β-catenin and Ras could be an ideal therapy for treating metastatic colorectal cancer
Authors: Cho Yong-Hee , Yoon Jeong-Soo , Choi Kang-Yell
Published: July 2017

INTRODUCTION Aberrant activations of Wnt/β-catenin and Ras signaling pathways by APC (90%) and K-Ras (40-50%) mutations are closely interacted and accelerate CRC tumorigenesis. Protein level of β-catenin as well as Ras is increased by APC loss via inhibiting GSK3B-mediated poly-ubiquitination dependent Ras degradation through recruitment of β-TrCP E3 linker. Moreover, additional K-Ras mutation results in the liver metastasis by secondary activation of the Wnt/β-catenin signaling pathways via Ras-ERK and -AKT pathways in addition to the initial activation by APC loss. Consequently, both β-catenin and Ras were highly increased in human CRC patient tissues, especially in metastatic tumor.

EXPERIMENT PROCEDURES To investigate the synergistic effects of aberrant Wnt/β-catenin and Ras signaling pathways on tumor development to metastasis, we established the APCMin/+/K-RasG12DLA2 compound mouse that harbor APC and K-Ras mutations in animal model. We also analyzed Ras protein level using human CRC patient tissues of normal mucosa and adenocarcinoma (N=24) and metastatic adenocarcinoma (N=26) and tumor budding (N=10). To recapitulate the metastatic tumor environment which have enriched receptor tyrosine kinases (RTKs), we maintained and experimented with EGF (20ng/ml) in every in vitro studies.

RESULTS We observed that both β-catenin and Ras were increased in adenocarcinoma and metastatic adenocarcinoma of human CRC compared with normal mucosa. Interestingly, both β-catenin and Ras were most significantly increased in tumor budding regions which histopathologically represent epithelial mesenchymal transition (EMT). In murine model, the EMT phenomenon was effectively induced and invasion of small intestinal tumors were occurred in APCMin/+/K-RasG12DLA2 with increased level of β-catenin and Ras compared with APCMin/+. However, EMT phenomenon and its following events such as migration and invasion of CRC cells induced by APC and K-Ras mutations were abolished by destabilization of β-catenin and Ras.

CONCLUSION Our results show that destabilizing β-catenin and Ras could be effective therapeutic targets for inhibiting metastasis in the early stage of CRC development

Citation Format: Yong-Hee Cho, Jeong-Soo Yoon, Kang-Yell Choi. Destabilizing β-catenin and Ras could be an ideal therapy for treating metastatic colorectal 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 911. doi:10.1158/1538-7445.AM2017-911
Abstract 912: ADAM9-plectin interaction involved in prostate cancer malignancy progression
Authors: Liu Che-Ming , Hsieh Chia-Ling , Sung Shian-Ying
Published: July 2017

Introduction: To identify a specific biomarker for malignant prostate cancer progression and therapeutic target, we had demonstrated the correlation of ADAM9 expression and cancer progression. The aim is to clarify the association of ADAM9 and interactive proteins in malignant phenotypic progression.

Methods: Knockdown of ADAM9 expression was conducted to explore its role in cell motility and metastasis activities. Pull-down assays were performed to identify the proteins that interact with ADAM9 during cancer cell migration. Proximity ligation assay (PLA) was carried out to validate the protein-protein interaction. Transmission electron microscope (TEM) analysis to confirm ADAM9 and plectin interaction. Protein degradation test was conducted to determine the regulation pathway.

Results: Our study demonstrated that ADAM9 play an important role in the regulation of prostate cancer migration, invasion and metastasis in the in vitro assay and xenograft animal models. The combination of ADAM9 pull-down assays and mass spectrum analysis demonstrated hemidesmosome components, plectin, ladinin, and cytoskeleton were pull-down with ADAM9. However, the key element of hemidesmosome, integrin α6β4, did not pull-down together. Both TEM and PLA confirmed the interaction of ADAM9 and plectin in the protrusion end of migrating cells. This interaction can be strengthened after knockdown integrin β4; and vice verse, knockdown of ADAM9 increased interaction of integrin β4 and plecin. These results indicated ADAM9 and integrin β4 compete for plectin binding. Furthermore, shADAM9 attenuated the degradation of integrin β4. Clinical evaluation of 96 patient samples in TMU demonstrated increased of ADAM9 and plectin interaction in tumor region when compared to the benign part of the same patient.

Conclusions: Our studies demonstrated that ADAM9 promotes cancer metastasis through alteration of hemidesmosome dynamic by enhancing of integrin β4 degradation. Therefore, these results indicated possible to use ADAM9-plectin interaction to predict prostate cancer malignancy progression.

Citation Format: Che-Ming Liu, Chia-Ling Hsieh, Shian-Ying Sung. ADAM9-plectin interaction involved in prostate cancer malignancy progression [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 912. doi:10.1158/1538-7445.AM2017-912
Abstract 917: CD82/KAI1 maintains the dormancy of long-term hematopoietic stem cells through interaction with DARC-expressing macrophages
Authors: Choi Jae-Il , Hur Jin , Lee Hwan , Nham Pniel , Chae Cheong-Whan , Choi Young-Eun , Kim Taewan , Lee Ga-Young , Baek Sung Hee , Kim Hyo-Soo
Published: July 2017

Hematopoiesis is regulated by crosstalk between long-term repopulating hematopoietic stem cells (LT-HSCs) and supporting niche cells in the bone marrow (BM). Here, we examine the role of CD82/KAI1 (CD82 hereafter) in niche-mediated LT-HSC maintenance. We found that CD82 is expressed predominantly on LT-HSCs and rarely on other hematopoietic stem-progenitor cells (HSPCs). In Cd82-/- mice, LT-HSCs were selectively lost as they exited from quiescence and differentiated. Mechanistically, CD82-based TGF-b1/Smad3 signaling leads to induction of CDK inhibitors and cell-cycle inhibition. The CD82 binding partner DARC/CD234 is expressed on macrophages and stabilizes CD82 on LT-HSCs, promoting their quiescence. When DARC+ BM macrophages were ablated, the level of surface CD82 on LT-HSCs decreased, leading to cell-cycle entry, proliferation, and differentiation. A similar interaction appears to be relevant for human HSPCs. Thus, CD82 is a functional surface marker of LT-HSCs that maintains quiescence through interaction with DARC-expressing macrophages in the BM stem cell niche. Finally, we hope that further development of our study will allow us to improve leukemia treatment by awakening not only “good” stem cells to keep beneficial stem cell functions, but also “bad” stem cells (e.g. cancer stem cells) in order to increase the sensitivity to treatment and avoid relapse.

Citation Format: Jae-Il Choi, Jin Hur, Hwan Lee, Pniel Nham, Cheong-Whan Chae, Young-Eun Choi, Taewan Kim, Ga-Young Lee, Sung Hee Baek, Hyo-Soo Kim. CD82/KAI1 maintains the dormancy of long-term hematopoietic stem cells through interaction with DARC-expressing macrophages [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 917. doi:10.1158/1538-7445.AM2017-917
Abstract 918: Cdc42 is crucial for intestinal stem cells survival by regulating Wnt and YAP signaling
Authors: Zhang Xiao , Gao Nan
Published: July 2017

Strictly defined architecture of intestinal epithelial cells and the gradient of signaling cues along the villus-crypt axis ensure the maintenance of adult intestinal stem cells, as well as proper regeneration and differentiation of all intestinal epithelial cell types. The Rho family small GTPase Cdc42 controls the polarized division of intestinal stem cells, and affects subsequent fate-determination of the progenitor cells. As we previously described in Cdc42 Villin-Cre knockout mice, loss of Paneth cells from crypt bottom and emergence of Goblet cell-like intermediate cells at the villus indicate possible compromise of the canonical Wnt signaling during Paneth cell differentiation. Here we further characterized the underlying molecular mechanism of Cdc42 regulating intestinal stem cell survival, proliferation, and differentiation. IHC of the intestinal tissue in Cdc42 Villin-Cre knockout mice suggests greatly reduced Olfm4 level in the stem cells, which are abnormally positioned in the crypts, along with a reduction of both transcription co-factors beta-Catenin and YAP. However, the Notch signaling, which is required for stem cells and the differentiation of absorptive linage, seems intact. The autonomous regulation of Cdc42 on Yap signaling is probably through its specific role in stabilizing cell-cell junction and actin filaments in the epithelium, both of which are known regulatory inputs of the Hippo-YAP pathway. Both Wnt/beta-Catenin and Yap signaling are required for the initiation and progression of colorectal cancer, and the regeneration of stem cells post radiation. Given that Cdc42 is overexpressed in multiple types of cancers, especially colorectal cancer, our finding that Cdc42 regulates both Wnt and YAP pathways in the homeostasis of adult intestinal tissue sheds new light on the molecular mechanism of its pathological involvement, and potential therapeutic target.

Citation Format: Xiao Zhang, Nan Gao. Cdc42 is crucial for intestinal stem cells survival by regulating Wnt and YAP signaling [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 918. doi:10.1158/1538-7445.AM2017-918
Abstract 926: Generation of a potential breast cancer stem cell model from induced pluripotent stem cells
Authors: Nair Neha , Calle Anna Sanchez , Zahra Maram Hussein , Oo Aung Ko Ko , Vaidyanath Arun , Masuda Shinobu , Kasai Tomonari , Seno Masaharu
Published: July 2017

The role of cancer stem cells (CSC) in the present scenario of breast cancer research is perceived as indispensable in the development and progression of the disease. The CSC niche maintains a heirarchy of heterogeneous cells, which facilitates the inception of the tumor until its invasion. Therefore, our study focuses on the development of an effective breast cancer stem cell model from mouse induced pluripotent stem cells (miPSCs).

Taking into consideration the intertumor heterogeneity of breast cancer, three human breast cancer cell lines, namely BT549, SKBR3 and T47D representing three major hormone subtypes were used to derive respective conditioned medium. The miPSCs were treated with these conditioned media separately for a period of one month following the protocol previously established by our laboratory. The resulting survived cells were subcutaneously and orthotopically transplanted into Balb/c nude mice. Within a short span of 15-20 days, both subcutaneous and orthotopic tumors were developed. Serial transplantations in to nude mice also generated malignant tumors with same vigor. Histopathological studies of these tumors confirmed a malignant infiltrating ductal carcinoma with apparent desmoplasia in orthotopic tumor and Her2 expression in comparison with subcutaneous ones, all of which confirmed us the conversion of miPSCs to CSCs. Presence of lung metastatic nodes further ascertained the invasiveness of this in vitro generated CSCs. Marked expression of prominent CSC markers namely CD44, Nanog, Sox2 and CD49f were observed in primary cells generated from these tumors. Tumor sphere formation assay further confirmed the presence of a good population of self renewing CSC population in the tumor tissues.

This study attempts to recapitulate human breast cancer disease in mice without any genetic manipulation, but by exploiting the tumor microenvironment in the form of conditioned medium. This should pave the way for the establishment of personalized therapy and a model to assess effective therapeutic interventions.

Citation Format: Neha Nair, Anna Sanchez Calle, Maram Hussein Zahra, Aung Ko Ko Oo, Arun Vaidyanath, Shinobu Masuda, Tomonari Kasai, Masaharu Seno. Generation of a potential breast cancer stem cell model from induced pluripotent stem 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 926. doi:10.1158/1538-7445.AM2017-926
Abstract 928: Combined treatment by manuka honey and metformin inhibited growth and inducted apoptosis in CD133+and CD90+ subpopulation hepatocellular carcinoma cancer stem cells
Authors: Fyala Amira S. , Sultan Ahmed S.
Published: July 2017

Manuka honey is well known for its anti-bacterial and wound healing properties but its anticancer role against cancer stem cells (CSC) in hepatocellular carcinoma (HCC) cell lines and the underlying molecular mechanisms remain uninvestigated. Recently, we identified a CSC subpopulation in HCC cell lines characterized by their CD133+ and CD90+ phenotype. The CD133+/ CD90+cells demonstrated a more aggressive phenotype than the CD133-/ CD90- counterpart and formed metastatic lesions in the lung of immunodeficient mice. In order to evaluate the effect of Manuka honey alone or in combined treatment with Metformin, a biguanide that has been widely used to treat type 2 diabetes, on cell viability and apoptosis induction in human HCC cell lines, isolated and identified CD133+/ CD90+subpopulation CSC were used. Cell viability was monitored using an MTT assay. In addition, apoptosis induction was labeled by Annexin V-fluorescein isothiocyanate/propidium iodide and measured using flow cytometry. Western blotting was used to examine the protein expression of p53, Bcl-2, Bax, Bcl-2-associated death promoter (Bad), Bcl-2 homologous antagonist/killer (Bak) and cleaved-caspases-3 and -9. The results of the present study revealed that Manuka honey alone or in combined treatment with Metformin suppressed the cell viability of CD133+/CD90+cells in a concentration-dependent manner and the maximum effect was detected in combined treatment after 24 h. In addition, Manuka honey alone or in combined treatment with Metformin induced a significant apoptosis in the tested cells. Furthermore, it was demonstrated that combined treatment increased the expression of proapoptotic proteins, including p53, Bax, Bad and Bak, while it decreased the protein level of antiapoptotic protein Bcl-2 compared to control, Manuka honey or Metformin alone. Furthermore, the combined treatment strongly inhibited stemness characteristics of CSC subpopulation, colony formation and dramatically inhibited CD133+/CD90+tumor growth in vivo. Our data demonstrated that combined treatment induced apoptosis in CD133+/CD90+ subpopulation through G1 arrest in the cell-cycle and regulating p53/Bcl-2/caspase-3 signaling pathway. In conclusion, our results suggested that Monuka honey alone or combined treatment with Metformin may be a potential treatment for apoptosis induction in CSC and may provide a novel therapeutic model for HCC.

Citation Format: Amira S. Fyala, Ahmed S. Sultan. Combined treatment by manuka honey and metformin inhibited growth and inducted apoptosis in CD133+and CD90+ subpopulation hepatocellular carcinoma cancer stem 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 928. doi:10.1158/1538-7445.AM2017-928
Abstract 929: Dissecting heterogeneity and population dynamics in breast cancer stem cells
Authors: Brooks Michael D. , Wicha Max S.
Published: July 2017

The CSC hypothesis posits that there is a subset of tumors cells driving tumor growth and metastasis and that by targeting this population, tumors can be more efficiently eradicated. Recently it was shown that there are two types of CSCs in breast cancer, mesenchymal CSC’s characterized as CD44+/CD24- and an epithelial type marked by ALDH expression. The cellular plasticity of these CSC states may contribute to their metastatic capacity. However, little is known about the heterogeneity of these CSC populations or how they are regulated by the tumor microenvironment. To address these issues we have utilized Drop-Seq to generate single cell RNA-Seq data from thousands of cells across patient derived xenografts from various molecular subtypes of breast cancer. In order to monitor real-time state transitions between the determined cellular populations, we have also designed and constructed fluorescent reporter constructs for genes specific to critical cell populations and cancer stem cells. We used CRISPR/Cas9 to introduce these reporters into the native loci of the genes in order to recapitulate native gene expression patterns. Single cell transcriptomics will help to characterize the molecular heterogeneity of CSC populations. The addition of fluorescent reporters will help to elucidate the mechanisms that regulate breast CSC’s as well as their interaction with the tumor microenvironment and to identify potential therapeutic targets.

Citation Format: Michael D. Brooks, Max S. Wicha. Dissecting heterogeneity and population dynamics in breast cancer stem 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 929. doi:10.1158/1538-7445.AM2017-929
Abstract 930: Role of DLK1 in sarcoma stem cells
Authors: Han Ilkyu , Ahn Sun Hee , Kim Han-Soo
Published: July 2017

Sarcoma is a rare form of cancer that comprises about 1% of all malignancies. A large proportion of deaths by sarcomas are caused by tumor metastasis, of which the biological basis remains obscure. A significant body of evidence suggests that only a small population of sarcoma cells possess the ability to initiate and maintain tumors. These tumor-initiating cells have the ability to self-renew, recapitulating the cellular heterogeneity of the original cancer, and thus are often referred as cancer stem like cells (CSLCs).Delta-like factor 1 (DLK1) was cloned in the preadipocyte cells 3T3-L1 cDNA library. Previous studies reported on the role of DLK1 in mesenchymal stem cell differentiation, adipogenesis, myogenesis, and osteoblastogenesis. It was reported that DLK1+ liver cancer cells have characteristics similar to those of CSLCs and that DLK1 could be a potential therapeutic target against the liver CSLCs. However, little is known about role of DLK1 in CSLCs of sarcoma. The mRNA (real time PCR) and protein (Western blot) expression of DLK1 from 13 sarcoma cell lines was examined. CSLCs from sarcoma cell lines were isolated using FACS with CD133 and sphere forming assay. The effect of DLK1 on sarcoma tumorigenesis was examined using cell proliferation and cell invasion assays by DLK1 siRNA treatment.Real-time PCR showed the increased expression of DLK1 mRNA in sarcoma cell lines. In particular, DLK1 mRNA was highly expressed in U2OS (osteosarcoma), SW872 (liposarcoma), A204 (rhabdomyosarcoma), and HS-SYII (synovial sarcoma) cells. We observed a reduction in cell proliferation and invasion in U2OS, A204, and sw872 cells treated with DLK1 siRNA. CSLCs were isolated from U2OS, A204, and sw872 by the FACS sorting method using the CD133, a CSLC marker in sarcoma. The number of active sarcoshperes in CD 133+ cells were significantly higher than that in CD133- cells by sphere forming assay. Soft agar assays revealed the presence of many more colonies in CD133+ cells than in CD133- or wild type cells. To investigate the effect of DLK1 on sarcoma CSLCs, sw872 cells were categorized into four groups: wild type, CD133-, CD133+ and spheres. Real-time PCR and western blot were used to detect the DLK1 gene in all four groups. We observed that DLK1 expression in sw872 CSLCs (CD133+ and spheres) were relatively low. We found that suppression of DLK1 by siRNA inhibited the invasiveness of CSLCs although it inhibited more effectively to activation of invasive cells as well as cell proliferation in sw872 non-CSLCs.These results suggest that DLK1 may play a role in tumorigenesis of CSLCs and cancer cells in various sarcomas.

Citation Format: Ilkyu Han, Sun Hee Ahn, Han-Soo Kim. Role of DLK1 in sarcoma stem 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 930. doi:10.1158/1538-7445.AM2017-930
Abstract 935: CD133+ cancer stem cells promoted by VEGF accelerate the recurrence of hepatocellular carcinoma
Authors: Chen Dexi , Liu Kai , Hao Meijun , Ouyang Yabo , Zheng Jiasheng , Challagundla Kishore Babu.
Published: July 2017

The function of cancer stem cells (CSCs) on inducing HCC recurrence after radiofrequency ablation (RFA) is still unclear. Here, 19 RFA-treated primary HCC patients were enrolled. We identified a dramatic increase of plasma VEGF in some patients (termed type II, n=9) after RFA, who suffered early HCC recurrence; the other patients (termed type I, n=10) had no increased plasma VEGF but had longer interval for HCC recurrence. Moreover, the expression of CSCs marker, CD133, was dramatically increased in recurrent HCC tissue of Type II in comparison with type I, suggesting induction of CD133+ CSCs may contribute to early HCC recurrence. In vitro studies demonstrated that VEGF stimulus enhanced the level of CD133+ CSCs and their self-renewal ability by inducing Nanog dependently on activation of VEGFR2. In vivo studies further demonstrated that CD133+ CSCs with VEGF stimulus formed larger tumor size in comparison with non-stimulus; and VEGF stimulus increased the tumorigenic cell frequency of primary HCC cells dependently on the presence of Nanog and VEGFR2. In recurrent HCC tissue of type II, but not type I, almost all CD133+ cells were Nanog and p-VEGFR2 positive cells, suggesting that activation VEGFR2 is critical for RFA-induced tumor stemness in HCC. Taken together, RFA-induced VEGF promotes tumor stemness and accelerates tumorigenesis in HCC dependently on Nanog and VEGFR2, which is valuable for the prediction of HCC recurrence after RFA and development of novel therapeutics.

Citation Format: Dexi Chen, Kai Liu, Meijun Hao, Yabo Ouyang, Jiasheng Zheng, Kishore Babu. Challagundla. CD133+ cancer stem cells promoted by VEGF accelerate the recurrence of hepatocellular carcinoma [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 935. doi:10.1158/1538-7445.AM2017-935
Abstract 937: Plasticity in heterogenous cancer stem cells promotes glioblastoma radioresistance
Authors: Minata Mutsuko , Audia Alessandra , Shi Junfeng , Lu Songjian , Pavlyukov Marat , Gillespie Yancey Gillespie , Bhat Krishna , Nakano Ichiro
Published: July 2017

Some cancers including Glioblastoma (GBM) have hierarchical cellular architecture with cancer stem cells (CSCs) as a minor tumorigenic subpopulation. While CSCs have been touted to contribute to initiate recurrence, post-surgical therapy-induced phenotypic changes in CSCs remain elusive. Here we report that treatment of GBM cells with ionizing radiation (IR) promotes a wound-healing mesenchymal signature, accompanied by conversion of naïve CSCs expressing CD133 to post-treatment more radioresistant CSCs expressing the GPI-anchored, TGFb-inhibitory protein CD109 in an ATM, NF-kB, CEBP-b dependent manner. In post-IR GBM, these therapy-escaped CD133-/CD109+ GBM cells, but not the rest of the tumor, uniquely retain clonogenic, tumorigenic, and multi-potent capacities both in vitro and in vivo. CD109 silencing unexpectedly eliminates cancer stemness via controlling YAP/TAZ as the downstream target and this CD109-YAP/TAZ signaling axis is evolutionally conserved between drosophila and human brain tumors. Clinically, both CD133 and CD109 are independently associated with poorer prognosis of GBM patients, while the poorest prognosis group of GBM expressed both CD133 and CD109 at higher rates. These data indicate a possibility of the post-IR CSC plasticity and subsequent heterogeneity, and the activation of the CD109-YAP/TAZ signaling axis as previously unidentified therapeutic targets for post-IR recurrent GBM.

Citation Format: Mutsuko Minata, Alessandra Audia, Junfeng Shi, Songjian Lu, Marat Pavlyukov, Yancey Gillespie Gillespie, Krishna Bhat, Ichiro Nakano. Plasticity in heterogenous cancer stem cells promotes glioblastoma radioresistance [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 937. doi:10.1158/1538-7445.AM2017-937