bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1629: Prednisone versus dexamethasone acute toxicity and cumulative doses variations in childhood acute lymphoblastic leukemia
Authors: Marcoux Sophie , Chapdelaine Aurélie , Robaey Philippe , Sinnett Daniel , Krajinovic Maja , Laverdière Caroline
Published: August 2015

INTRODUCTION

Glucocorticoids (GC) are the main components of childhood acute lymphoblastic leukemia (ALL) treatment; in the past decades, there were numerous attempts at finding whether prednisone or dexamethasone is the best drug.

EXPERIMENTAL PROCEDURE

A retrospective study of patients (n = 119) treated according to Dana-Farber Cancer Institute (DFCI)-ALL treatment protocols 91-01 and 95-01 was done. Medical charts were reviewed and type and doses of glucocorticoids (GC) received were recorded. Bone and psychiatric symptoms, as well as all anthropomorphic data were also recorded.

For each patient, a total theoretical dose was calculated according to the initial risk of relapse stratification, treatment protocol indications and assigned arms. Cumulative GC doses actually received were compared to theoretical doses in the form of a dose deviation in% ((theoretical dose - actual dose) / theoretical dose * 100).

SUMMARY OF DATA

Results indicate that despite an adequate correlation between planned and administered GC doses for most patients, patients treated with dexamethasone were found to have significantly more GC dose deviations than those treated with prednisone (p < 0.001). Findings also indicate that bone toxicity (p = 0.02), but not psychiatric toxicity, is associated with GC dose deviations.

CONCLUSIONS

Despite representing the current standard in clinical studies, using only different treatment arms or different treatment protocols may lead to significant interpretation caveats if doses actually received are not controlled for. This is of importance especially for drugs likely to induce very severe side-effects requiring dose modification, as it is the case for GC and acute bone toxicity.

Citation Format: Sophie Marcoux, Aurélie Chapdelaine, Philippe Robaey, Daniel Sinnett, Maja Krajinovic, Caroline Laverdière. Prednisone versus dexamethasone acute toxicity and cumulative doses variations in childhood acute lymphoblastic leukemia. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1629. doi:10.1158/1538-7445.AM2015-1629
Abstract 1679: Immunotherapeutic strategies targeting CXCR4 axis plus γ/δ T cells in treating ovarian cancer
Authors: Liu Chao Lien , Mao Tsui Lien
Published: August 2015

Ovarian carcinoma is the most crucial and difficult problem for therapeutic treatment in gynecological malignancies, which requires great efforts for an effective solution. Molecular studies have shown that chemokine stromal cell-derived factor-1 (SDF-1)/CXCL12 and its receptor CXCR4 are the key determinants of tumor initiation, progression and metastasis in ovarian carcinoma. It is also believed that blocking CXCR4-CXCL12 pathway plays potential roles in treating ovarian cancer patients. Recent studies also showed that the cytotoxic function of gamma delta (γ/δ) T lymphocytes could efficiently kill ovarian CSCs (cancer stem cells) and present a promising immunotherapy for ovarian cancer. To investigate how the CXCL12/CXCR4 pathway is involved in the regulation of ovarian cancer. We eliminated the expression of CXCR4 in different ovarian cancer cell lines using CXCR4 antagonist (e.g., AMD3100) and CXCR4 shRNA silencing to prevent the CXCR4/CXCL12 axis interaction. Besides, up to 78% of γ/δ T cells was expanded in-vitro using aminobisphosphonates plus low-dose interleukin-2 for the therapeutic investigating. The therapeutic effects following different treatments were measured. First, our results showed that blocking of Cxcr4/Cxcl12 axis by AMD3100 reduced around 37% on cell survival rate. Similar results were also observed in the CXCR4-shRNA silencing ovarian cancer cells. Additionally, the higher CXCR4-expressed cells were highly proliferation after CXCL12 stimulating compared to the CXCR4 non-expressed cells. Next, cancer cells treated with the exogenous activated γ/δ T cells showed a reduction of 35% on cell survival rate. Finally, the combined treatment strategy using AMD3100 plus γ/δ-T cells showed the most significant therapeutic effects in the high-grade ovarian cancer cells, and the cell survival rate was reduced almost to 50% after the cell cytotoxic assay. We therefore conclude that the limitation of CXCL12/CXCR4 activities combined with the γ/δ T cell cytotoxicity is a potent immunotherapy for treating ovarian cancer and would served as a promising new strategies for clinical translation.

Citation Format: Chao Lien Liu, Tsui Lien Mao. Immunotherapeutic strategies targeting CXCR4 axis plus γ/δ T cells in treating ovarian cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1679. doi:10.1158/1538-7445.AM2015-1679
Abstract 1716: Acquisition of estrogen independence induces TOB1-related mechanisms supporting breast cancer cell proliferation
Authors: Zhang Yong-Wei , Nasto Rochelle E. , Varghese Rency , Jablonski Sandra A. , Serebriiskii Ilya G. , Surana Rishi , Calvert Valerie S. , Bebu Ionut , Murray Joseph , Jin Lu , Johnson Michael , Riggins Rebecca , Ressom Habtom , Petricoin III Emmanuel , Clarke Robert , Golemis Erica A. , Weiner Louis M.
Published: August 2015

Approximately 70% of newly diagnosed breast cancers express high levels of estrogen receptor alpha (ERα). However, de novo and acquired drug resistance to anti-estrogen agents limit the treatment of such patients, requiring the use of cytotoxic chemotherapy; many such patients have a poor prognosis. Therefore, resistance to therapies targeting the estrogen pathway remains a challenge in the treatment of estrogen-receptor positive breast cancer. To address this challenge, a system biological approach by screening a library of siRNAs targeting an estrogen receptor- and aromatase-centered network, was used and identified 46 genes that are dispensable in estrogen-dependent MCF7 cells, but are selectively required for the survival of estrogen-independent MCF7-derived cells, and multiple additional estrogen-independent breast cancer cell lines. Integration of this information identified a tumor suppressor gene TOB1 as a critical determinant of estrogen-independent estrogen-receptor positive breast cell survival. Depletion of TOB1 selectively promoted G1 phase arrest and sensitivity to AKT and mTOR inhibitors in estrogen-independent cells but not estrogen-dependent cells. Phosphoproteomic profiles (results from reverse phase protein array, RPPA) under basal conditions and following TOB1 depletion identified significant signaling network reprogramming by TOB1 that differed in estrogen-sensitive and estrogen-resistant cell lines. These data support a novel function for TOB1 in mediating survival of estrogen-independent breast cancers. The translational significance of this new molecular mechanism of estrogen independence in human breast cancer will be fully determined when TOB1 inhibitors are developed to enable in vivo validation studies. These studies also provide evidence for combining TOB1 inhibition and AKT/mTOR inhibition as a therapeutic strategy, which have potential translational significance for the management of patients with estrogen receptor-positive breast cancers.

Citation Format: Yong-Wei Zhang, Rochelle E. Nasto, Rency Varghese, Sandra A. Jablonski, Ilya G. Serebriiskii, Rishi Surana, Valerie S. Calvert, Ionut Bebu, Joseph Murray, Lu Jin, Michael Johnson, Rebecca Riggins, Habtom Ressom, Emmanuel Petricoin III, Robert Clarke, Erica A. Golemis, Louis M. Weiner. Acquisition of estrogen independence induces TOB1-related mechanisms supporting breast cancer cell proliferation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1716. doi:10.1158/1538-7445.AM2015-1716
Abstract 1768: Chloroquine potentiates carfilzomib but not bortezomib effects on myeloma cells
Authors: Misund Kristine , Baranowska Katarzyna Anna , Holien Toril , Starheim Kristian , Johansson Ida , Buene Glenn , Waage Anders , Bjørkøy Geir , Sundan Anders
Published: August 2015

Proteasomal inhibition has proven good clinical effect in Multiple Myeloma; however, some patients are resistant to treatment and most patients acquire resistance during the disease course. It is well known that cells degrade proteins by two distinct but interconnected pathways; one is protein degradation by proteasomes, which may be clinically targeted by proteasome inhibitors, the other is by formation of autophagosomes and lysosomal degradation, which can be inhibited by hydroxychloroquine (HCQ). Assuming that the two arms of protein degradation may compensate for each other, there is a rationale to simultaneously target both arms. We tested the combined effects of HCQ and the reversible proteasome inhibitor bortezomib or the irreversible inhibitor carfilzomib on myeloma cell lines and primary cells. As expected, HCQ potentiated carfilzomib-induced myeloma cell death, but surprisingly, HCQ had little or no effect on bortezomib activity. We also found that HCQ potentiated the effect of the irreversible proteasome inhibitor oprozomib, but not the effect of the reversible inhibitor ixazomib. Thus, the sensitivity towards the irreversible proteasome inhibitors carfilzomib or oprozomib is to a larger extent affected by the autophagic system than the reversible inhibitors bortezomib and ixazomib. In conclusion, we here suggest that in attempts to increase the clinical efficacy of proteasome inhibitors, inhibitors of autophagy and lysosomal degradation such as HCQ should be combined with an irreversible proteasome inhibitor such as carfilzomib, and not with bortezomib.

Citation Format: Kristine Misund, Katarzyna Anna Baranowska, Toril Holien, Kristian Starheim, Ida Johansson, Glenn Buene, Anders Waage, Geir Bjørkøy, Anders Sundan. Chloroquine potentiates carfilzomib but not bortezomib effects on myeloma cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1768. doi:10.1158/1538-7445.AM2015-1768
Abstract 1840: Metabolomic profiling of gastrointestinal stromal tumor (GIST) T1 cell lines in response to imatinib therapy
Authors: Marks Vered , Tiesi Gregory , Paz Mejia Ana , Trent Jonathan , Walls Jamie , Livingstone Alan , Yakoub Danny
Published: August 2015

Background: There is a growing need for novel drugs to treat drug resistant GIST. The aim of this study was to evaluate chronological metabolic changes in GIST cells treated with imatinib mesylate.

Methods: Human GIST T1 cells were incubated with imatinib 0.5 mM. Metabolomic profiling was performed in extracted cell pellets at 12, 24 and 48 hours, after viability and cell counting was done, and compared with controls via acquisition of 1H-NMR spectra using a 500MHz spectrometer equipped with a 5mm TCI 500S2 H-C/N-D-05 Z cryoprobe head at 298 K. Standard, one-dimensional NOESYpr1D pre-saturation pulse program was used. Spectra were further processed in an NMR suite processor; Partial Least Squares Discriminant Analysis (PLS-DA) model was used for characterization of chronological biochemical differences.

Results:

By 48 hours of exposure of cells to imatinib, glucose initially increased then significantly decreased with gradual decrease of lactate production and minimal changes in pyruvate and succinate denoting shifting from cytosolic to mitochondrial glycolysis, this was echoed with consumption of glutamine and glutamate. Glutathione and cell membrane phospholipids as phosphocholine gradually decreased by a factor of 1.6 for each, suggesting inhibited growth and invasiveness. Aspartate, myo-inositol, glycerophosphocholine and taurine gradually increased by a factor of 1.6, 1.7, 2.6 and 1.3, respectively. Tyrosine, valine and leucine showed minimal changes. Cell viability was directly correlated to changes in choline, creatine phosphate, myo-inositol and taurine (Pearson's R >0.65). PLS-DA model suggested that changes in myoinositol, glycerophosphocholine, glutamate, aspartate, phosphcholine and glutathione were the metabolites mostly influencing the differential chronological global metabolic profiles of the cells with continued exposure to imatinib (R2X = 0.76, R2Y = 0.60, Q2(cum) = 0.20).

Conclusion:

Metabolomic profiling of GIST cells exposed to signal transduction modulators supplements molecular findings and provides further mechanistic insights into longitudinal changes of the mitochondrial and glycolytic pathways of oncogenesis, it can potentially provide novel as well as complementary therapeutic targets.

Citation Format: Vered Marks, Gregory Tiesi, Ana Paz Mejia, Jonathan Trent, Jamie Walls, Alan Livingstone, Danny Yakoub. Metabolomic profiling of gastrointestinal stromal tumor (GIST) T1 cell lines in response to imatinib therapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1840. doi:10.1158/1538-7445.AM2015-1840
Abstract 2207: Elucidating mechanisms employed by NFIL3 to impact cancer cell survival
Authors: Keniry Megan , Caballero Omar , Flores Itzel , Salinas Andrea
Published: August 2015

NFIL3 was recently identified as a novel modulator of FOXO1 output on the PI3K pathway. NFIL3 bound to DNA to physically block FOXO1 recruitment to genes that induce apoptosis. The diminishment in NFIL3 expression via RNA interference led to FOXO recruitment to cell death genes, an increase in Histone H3 acetylation on these genes, and increased expression of these genes. However, a number of observations suggest that FOXO1 is not the main target of NFIL3 in these cells. First, the NFIL3 and FOXO1 DNA consensus sites have almost no similarity. Second, the loss of FOXO1 or all three FOXO factors failed to suppress the cell death phenotype upon the diminishment of NFIL3 in MEFs. The goal of this project is to identify the factor(s) that NFIL3 directly blocks from DNA recruitment to promote cellular survival. In previous work, the NFIL3 and FOXO1 sites were mapped as being adjacent on the DNA. The mapped NFIL3 site was required for NFIL3 to associate with the DNA in biochemical experiments and for NFIL3 to regulate the promoter sequence in reporter assays. The mapped NFIL3 site is a perfect match for the bZIP transcription factor CCAAT/enhancer binding protein beta (CEBPB) consensus sequence. NFIL3 has been shown to block the ability of CEBPB to activate transcription by competing for access to the same DNA consensus site in numerous contexts including neuronal regeneration, PKA pathway transcriptional programs and the regulation of hepatitis C viral genes. Aside from DBP in circadian rhythm, CEBPB is the most commonly antagonized factor by NFIL3 in the literature. We are investigating if NFIL3 hinders FOXO1 recruitment to cell death genes by directly antagonizing CEBPB recruitment to these genes using cell biological and bioinformatics approaches. We have found that CEBPB is required for efficient induction of a subset of NFIL3 repressed genes.

Citation Format: Megan Keniry, Omar Caballero, Itzel Flores, Andrea Salinas. Elucidating mechanisms employed by NFIL3 to impact cancer cell survival. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2207. doi:10.1158/1538-7445.AM2015-2207
Abstract 2208: Interferon Consensus Sequence-Binding Protein (ICSBP) regulates the transforming growth factor-beta type I receptor expression in osteosarcoma cells
Authors: Sung Jee Young , Kim Hyeryeong , Yoon Kyung-Sil , Kim Yong-Nyun , Park Byung-Kiu
Published: August 2015

Transforming growth factor-beta (TGF-β), known as a tumor suppressor, may exert tumor promoting activity in the advanced stage of cancer. Previously, we reported that interferon consensus sequence-binding protein (ICSBP), also known as interferon regulatory factor-8, up-regulates TGF-β type I receptor (TGF-βR I), and augments growth in HL-60 leukemic cells. In the current study, we demonstrate that ICSBP expression up-regulates TGF-βR I and increase in cell growth in U2OS human osteosarcoma cells. To further investigate the mechanism of TGF-βR I up-regulation by ICSBP, we tested the possibility that ICSBP regulates promoter activity of TGF-βR I gene. We analyzed the promoter region of the human TGF-βR I gene in U2OS cells to find putative transcription elements regulated by ICSBP. Luciferase reporter gene assays indicated that ICSBP expression could enhance TGF-βR I gene expression, and by promoter deletion analysis we identified a functional ICSBP-dependent region located at nucleotides -220 to +5 in the TGF-βR I promoter. We also performed EMSA (Electro Mobility Shift Assay) using probe and found that there are putative ICSBP binding sites between -222 to -204 regions.

In conclusion, ICSBP increased TGF-βR I expression by binding to TGF-βR I promoter (-220/218) regions in U2OS cells.

Note: This abstract was not presented at the meeting.

Citation Format: Jee Young Sung, Hyeryeong Kim, Kyung-Sil Yoon, Yong-Nyun Kim, Byung-Kiu Park. Interferon Consensus Sequence-Binding Protein (ICSBP) regulates the transforming growth factor-beta type I receptor expression in osteosarcoma cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2208. doi:10.1158/1538-7445.AM2015-2208
Abstract 2210: DNA methylation to identify non-coding gene regulatory regions in tumor samples
Authors: Berman Benjamin P.
Published: August 2015

Altered control of transcriptional regulation via mutations to transcription factors and chromatin modifiers is a hallmark of cancer. Many popular methods for studying transcriptional regulation, such as ChIP-seq, DNase-I hypersensitivity, and Chromatin Conformation Capture, can be challenging to apply to clinical samples. DNA methylation has long been the chromatin biomarker of choice in disease studies because of its stability under clinical collection conditions. We have used DNA methylation profiles of thousands of tumors within the TCGA to reconstruct cancer-specific transcription factor regulatory networks, including non-coding transcription factor binding sites in enhancer regions. We use methyl-expression QTL (M->E QTL) analysis to associate changes at individual binding sites with their gene targets, often hundreds of thousands of base pairs away. This new approach allows for the first time the identification of in vivo enhancer changes in thousands of individuals, and provides a technology platform to identify key regulatory events in cancer progression.

Note: This abstract was not presented at the meeting.

Citation Format: Benjamin P. Berman. DNA methylation to identify non-coding gene regulatory regions in tumor samples. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2210. doi:10.1158/1538-7445.AM2015-2210
Abstract 2211: A comparative analysis of the cis-regulatory landscape between human and mouse
Authors: Yue Feng , Shen Yin , Ye Zhen , Ren Bing
Published: August 2015

As the premier model organism in biomedical research, the laboratory mouse shares the vast majority of protein-coding genes with humans, but significant differences exist between the two mammals, posing considerable challenges in the study and modeling of human diseases. The mouse ENCODE consortium applied the same high throughput approaches as the human ENCODE studies and produced more than1000 coordinated datasets in over 100 mouse cell types and tissues and greatly expanded the annotation of the mouse genome. By comparative analysis with the human genome, we found that although the majority of gene expression and cis-regulatory elements are conserved between the two species, a large degree of gene regulatory elements appear to be species-specific and these species-specific elements are enriched for genes involved in immune system and metabolic process, suggesting that different gene pathways evolve at distinct rates. Moreover, we found that species-specific elements are also enriched for repetitive DNA elements such LTR and SINE, which we hypothesize to be responsible generating the species-specific elements. Finally, we developed a novel computational method to study the functional conservation between two species without requiring perfectly matched cell lines or tissues. The result confirms that cis-elements with different evolutionary rates are associated distinct biological pathways and reveals the complex driving forces in the evolution of cis-regulatory landscape in mammals.

Note: This abstract was not presented at the meeting.

Citation Format: Feng Yue, Yin Shen, Zhen Ye, Bing Ren. A comparative analysis of the cis-regulatory landscape between human and mouse. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2211. doi:10.1158/1538-7445.AM2015-2211
Abstract 2217: Prognostic and predictive values of CD68+ macrophage and aberrant expression of cathepsin L, IL-12, CTL4A genes in classic Hodgkin's lymphoma
Authors: Zekri Abdel-Rahman N. , Bahnassy Abeer A. , El-Bastawisi Ahmad E-S , Hammad Osama , Ali Nasr M. , Diks John E. , Yousif Hend F.
Published: August 2015

Background: Despite advances in the treatment of classic Hodgkin lymphoma (CHL), current therapies fail to cure 10%-15% of patients, and a similar proportion of patients may be over treated, resulting in both short-term and long-term treatment-related complications. Current prognostic models predict the outcome of treatment with imperfect accuracy, and clinically relevant biomarkers have not been established to improve on international prognostic score.

Patients and Methods: CD 68 protein and RNA expression were assessed in 81 classic Hodgkin's lymphoma patients from Egypt by immunohistochemistry and Quantitative real time PCR (Q-RTP) respectively. Then CD 68 positive and CD 68 negative cases where profiled using the SABioscience array (Qiagen). Genetic aberrations were correlated to standard prognostic factors, patients response to the treatment, progression free and overall survival (PFS, OS) rates.

Results: Increased CD68 protein and RNA expression were detected in 70.3% and 53.1% of the cases, respectively. Increased CD68 protein associated significantly with old age >40 (p = 0.035), advanced stage (p = 0.007), and poor response to treatment (DP/SD, p = 0.001). CD68 RNA expression significantly associated with age (p = 0.035), and poor response to treatment (DP/SD, p = 0.001). CD68+ positive patients, showed high expression of cathepsin L, IL-12, ATXN2L, CTL4A and MMP11. The 5 years OS correlated significantly with CD68, IL-12, MMP11 and CTL4A-RNA expression (p< 0.001) as well as with age (p<0.001) and stage (p<0.001). The 5 years PFS correlated significantly with age (p = 0.003), stage (p = 0.012), CD68 protein and RNA expression (p< 0.001) as well as with cathepsin L, IL-12, CTL4A and MMP11 (p<0.01). CD68 protein expression significantly correlated with 2 years OS (p< 0.001).

Conclusions: in classic Hodgkin's lymphoma, CD68+ macrophage and aberrant expression of cathepsin L, IL-12, CTL4A and MMP11 provide sensitive biomarkers that can accurately predict patients response to treatment and survival with high accuracy.

Citation Format: Abdel-Rahman N. Zekri, Abeer A. Bahnassy, Ahmad E-S El-Bastawisi, Osama Hammad, Nasr M. Ali, John E. Diks, Hend F. Yousif. Prognostic and predictive values of CD68+ macrophage and aberrant expression of cathepsin L, IL-12, CTL4A genes in classic Hodgkin's lymphoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2217. doi:10.1158/1538-7445.AM2015-2217
Abstract 2268: Tenascin C as an effector of prostate cancer derived bone metastasis
Authors: San Martin Rebeca , Rowley David R. , Pienta Kenneth J.
Published: August 2015

The stroma plays a key role in maintenance of tissue homeostasis. A reactive stroma evolves early in the progression of prostate cancer. One way reactive stroma remodels the microenvironment is by deposition of tenascin-C. Tenascin-C is an extracellular matrix protein that is expressed during development and is critical for neuronal patterning and osteogenesis. In contrast, expression of tenascin C in adult tissues is limited and is restricted to regions of wound repair, tissue remodeling and pathological conditions, such as cancer.

To assess whether the bone exhibits a reactive tissue phenotype in the context of metastasis, human prostate cancer metastasis tissue arrays were evaluated using immunohistochemistry and spectral deconvolution. This work identified a tenascin-C expression pattern at trabeculae-associated metastatic sites, suggesting the evolution of a reactive endosteum.

In order to evaluate the mechanisms involved, we developed an in vitro 3D osteogenic organoid, using human mesenchymal stem cells induced to osteoblastic differentiation, which exhibits a reactive endosteum phenotype. Co-culture with the metastatic prostate cancer cell line VCaP showed preferential binding at sites high in tenascin C deposition. Also, metastatic cells were capable of adherence to purified tenascin C in vitro, forming 3D colonies. We have identified α9β1 integrin as a mediator of prostate cancer cell adhesion to tenascin C-rich surfaces. Preliminary data has also identified signaling through WNK1 and STAT6 as additional candidate mechanisms that mediate tenascin-C induced biology in prostate cancer cells that are metastatic to bone surfaces.

These studies characterize a reactive endosteum phenotype at sites of metastatic prostate cancer foci and suggest that elevated tenascin-C at these sites mediates the adhesive and other biological properties of cancer cells. This study will help provide data from which to develop novel therapeutic approaches to treat metastatic disease.

Citation Format: Rebeca San Martin, David R. Rowley, Kenneth J. Pienta. Tenascin C as an effector of prostate cancer derived bone metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2268. doi:10.1158/1538-7445.AM2015-2268
Abstract 2271: Peptidylarginine deiminase 2 as a novel therapeutic target for breast cancer
Authors: Horibata Sachi , McElwee John L. , Sadegh David , Rogers Katherine , McLean Dalton , Coonrod Scott A.
Published: August 2015

Peptidylarginine deiminases (PADIs) enzymes are calcium-dependent enzymes that post-translationally convert positively charged protein arginine residues to a neutrally charged citrulline in a process known as citrullination or deimination. This loss of charge can alter the tertiary structure of proteins and affect protein-protein and protein-DNA interactions. We, and others, have found that PADI-mediated histone tail citrullination alters chromatin structure and regulates gene expression. For example, we found that PADI2 interacts with estrogen receptor (ERα) and that PADI2-mediated citrullination of histone H3 arginine 26 residues at ERα binding sites regulates target gene expression. In addition, we also found that PADI2 is often co-expressed with HER2 in both breast cancer cell lines and in tumor tissue. Our molecular studies suggest that PADI2 activates HER2 expression via histone citrullination at the HER2 gene locus and that HER2 signaling appears to induce PADI2 expression, thus potentially forming an oncogenic positive feedback loop with HER2. These findings suggest an important role for PADI2 in breast cancer progression. This prediction is supported by our finding that PADI2 expression is upregulated in cancer cells and that depletion of PADI2 from breast cancer cells results in reduced tumorigenicity. In addition, we found that transgenic overexpression of PADI2 promotes carcinogenesis and that the PADI inhibitor, Cl-Amidine, slows tumor growth in mouse models of breast cancer. Furthermore, we found that Cl-Amidine also maintains basement membrane integrity in xenograft tumors, preventing initiation of metastasis. Together, these findings suggest that PADI2 represents a novel therapeutic target and biomarker for early stage breast cancer.

Citation Format: Sachi Horibata, John L. McElwee, David Sadegh, Katherine Rogers, Dalton McLean, Scott A. Coonrod. Peptidylarginine deiminase 2 as a novel therapeutic target for breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2271. doi:10.1158/1538-7445.AM2015-2271
Abstract 2274: The dual role of fibronectin in lung cancer progression
Authors: Cheng Hung-Chi , Chang Ming-Min , Tseng Yau-Lin
Published: August 2015

Tumor cells chronically progress, involving a series of aberrant activities within distinct organs, to accomplish final metastasis. The role of fibronectin (FN) in tumor progression has been controversial, being either tumor-suppressive or metastatic-promoting. Nevertheless, little effort has been made to reconcile these discrepancies in a systemic manner. Here, we deprived metastatic tumor cells of pericellular FN (periFN) in the spontaneous metastasis assay and demonstrated a dual role of FN. We found that, while knocking down FN expression of Lewis lung carcinoma cells (FNKD-LLC) increased the tumor sizes and migratory activities, it significantly decreased the tumor metastasis in the lungs. Histological examination revealed that subcutaneous tumors of mice bearing FNKD LLC invaded the surrounding tissues to a higher extent than those of mice bearing control scramble-LLC. Immunohistochemical staining with antibodies against CD31, FN, and casepase 3, revealed that FNKD-LLC rendered more angiogenesis, tumor intravasation, and fibrous FN deposited in the surrounding inflammatory tissues, without altering the tumor apoptosis, suggesting that the FN expression in tumor cells indeed plays a role in suppressing the tumor progression in the primary tumor tissues. Conversely, the metastatic tumor nodule numbers in the lungs were significantly reduced for the FNKD-LLC cells, clearly indicative of a metastatic-promoting role of periFN. We reasoned these seemingly conflicting phenomena by hypothesizing that the FN expression of neoplastic lung carcinoma cells is initially downregulated to trigger fibrotic microenvironments (FN deposition in extracellular matrices) to which pro-inflammatory leukocytes are recruited, leading to the enhanced angiogenesis and tumor intravasation. However, the early circulating lung cancer cells fail to colonize distant organs (including lungs) until periFN is re-assembled in the later progression stages, thus facilitating the tumor cell adhesion to CD26 on endothelial cells and consequently extravasation and metastasis. This hypothesis of temporal-spatial regulations was clinically supported by comparing and statistically analyzing the FN expression levels in lung cancer tissues collected from patients hospitalized by the NCKU hospital for surgery within 2003 through 2006 with low and high survivals. This study was supported by Grant # MOST 102-2320-B-006 -031- and MOST 103-2325-B-006 -009 -.

Citation Format: Hung-Chi Cheng, Ming-Min Chang, Yau-Lin Tseng. The dual role of fibronectin in lung cancer progression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2274. doi:10.1158/1538-7445.AM2015-2274
Abstract 2278: The hypusine cascade promotes cancer progression and metastasis through the regulation of RhoA in squamous cell carcinoma
Authors: Muramatsu Tomoki , Kozaki Ken-ichi , Imoto Seiya , Yamaguchi Rui , Tsuda Hitoshi , Kawano Tatsuyuki , Morishita Maki , Miyano Satoru , Inazawa Johji
Published: August 2015

Metastasis is one of the critical factors contributing to poor prognosis in cancer, but the mechanisms are poorly understood. We established a highly metastatic subline (HOC313-LM) derived from an oral squamous cell carcinoma cell line (HOC313) for uncovering the metastatic mechanisms. We identified deoxyhypusine synthase (DHPS) as a metastasis-related gene through the specific amplification at 19p13.2-p13.13 in the HOC313-LM. Overexpression of DHPS promoted cell motility, invasion and metastasis through activation of the RhoA signaling pathway, and increased the tumor formation in vivo. We also demonstrated that eukaryotic translation factor 5A (eIF5A) bound to RhoA mRNA and regulated its translation. Moreover, GC7, a DHPS inhibitor, showed significant decrease in the tumor formation in vivo. In patients with esophageal squamous cell carcinoma (ESCC), overexpression of DHPS in ESCC tumors was significantly associated with worse recurrence-free survival, and correlated with distant metastasis. Our elucidation of these molecular mechanisms within the hypusine cascade suggests opportunities for novel therapeutic targets in SCC.

Citation Format: Tomoki Muramatsu, Ken-ichi Kozaki, Seiya Imoto, Rui Yamaguchi, Hitoshi Tsuda, Tatsuyuki Kawano, Maki Morishita, Satoru Miyano, Johji Inazawa. The hypusine cascade promotes cancer progression and metastasis through the regulation of RhoA in squamous cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2278. doi:10.1158/1538-7445.AM2015-2278
Abstract 2282: K-RasV12 bone marrow cell reconstitution causes lung cancer in wild type mice
Authors: Belloni Elena , Martin-Padura Ines , Gerbino Elvira , Orecchioni Stefania , Fusar-Imperatore Fulvia , Marighetti Paola , Bertalot Giovanni , Pelicci Pier Giuseppe , Bertolini Francesco
Published: August 2015

The potential of recruited bone-marrow-derived cells (BMDCs) to act as a source of malignancy was disputed for a long time. A role for BMDCs in tissue repair has also been proposed, since these cells are capable of homing to the lung and differentiating into multiple epithelial cell lineages so thus contributing in lung tissue repair and regeneration. We tested the ability of bone marrow cells containing an inducible oncogene to initiate lung adenocarcinoma. Since the role of the K-Ras gene in lung cancer is well known, for our experimental strategy, we used a mouse model expressing oncogenic K-RasV12 (K-Ras+/V12RERTert/ert). This model generates a specific neoplastic phenotype induced by the K-RasV12 mutation. Mice usually develop lung tumors with complete penetrance evolving from hyperplasia, through adenoma, to adenocarcinoma, when the oncogene expression is induced by 4-hydroxitamoxifen (4-OHT) administration. Following lethal irradiation, wild-type mice were transplanted with donor bone marrow cells derived from K-RasV12 mice and, once the bone marrow was reconstituted, treated with 4-OHT. Mice were monitored and sacrificed at subsequent post-induction time points (0, 3.5, 5.5, 8.5, 11, 13.5, 15, and 16 months) and lungs and peripheral blood were collected. Analysis of the H&E-stained lung sections from both reconstituted K-RasV12 and control mice revealed diffuse hyperplasia, possibly due to irradiation. Epithelial lesions, adenomas and/or adenocarcinomas, were detected in 4 out of 8 K-RasV12-reconstituted animals at 3.5, 5.5, 13.5 and 15 months post oncogene induction while no lesions were detected in the control group (transplanted but not induced mice). In addition, the K-Ras implication on the origin of the tumor was confirmed by X-Gal staining of lung lesions in reconstituted mice. In our model, -geo is indeed a marker protein coexpressed along with the same K-RasV12 transcript by a bicistronic strategy. Our results suggest a direct implication of BMDCs in lung epithelial tumor development and emphasize the need of further studies to dissect the mechanisms regulating BMDCs’ recruitment to injured tissues and their role in tumor initiation and/or progression.

Citation Format: Elena Belloni, Ines Martin-Padura, Elvira Gerbino, Stefania Orecchioni, Fulvia Fusar-Imperatore, Paola Marighetti, Giovanni Bertalot, Pier Giuseppe Pelicci, Francesco Bertolini. K-RasV12 bone marrow cell reconstitution causes lung cancer in wild type mice. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2282. doi:10.1158/1538-7445.AM2015-2282
Abstract 2289: A novel mouse model of endometrial cancer: epithelial specific Pten deletion with lactoferrin-iCre
Authors: Daikoku Takiko , Terakawa Jumpei , Dey Sudhansu K.
Published: August 2015

Endometrial cancer is a common gynecological malignancy, causing approximately 8,000 deaths each year in the United States. Underlying causes of endometrial cancer are not clearly understood, and treatment options for patients with advanced stages are limited. Animal models that spontaneously develop cancer are powerful tools for studying the mechanism of cancer initiation and progression and for developing treatment strategies. We previously created a mouse model of endometrial cancer in which the floxed phosphatase and tensin homologue (Pten) gene was deleted by Cre expression under the control of a progesterone receptor (PR) promoter. Pten is a tumor suppressor with the highest frequency of mutation in human endometrial cancer. In the PtenloxP/loxP/PR-Cre mouse model, uterine deletion of Pten resulted in complex atypical hyperplasia of the uterine epithelium by three weeks of age with cancer occurring by one month. As PR is expressed in the epithelium, stroma, and myometrium compartments, the question remains whether Pten deletion in the uterine epithelium alone contributes to endometrial cancer since human endometrial cancer is derived from epithelial cells. To address this question, we used lactoferrin-iCre (Ltf-Cre) mice to specifically delete Pten in the uterine epithelium, since lactoferrin is restricted to the epithelium of adult mice. We found that PtenloxP/loxP/Ltf-Cre mice develop complex atypical hyperplasia in uterine epithelium by three months of age, whereas uteri from immature mice are normal. These results demonstrate that Pten deletion specifically in the uterine epithelium can contribute to toward endometrial cancer in mice. This mouse model will help us to study in detail the initiation and progression of endometrial cancer. This study was supported in parts by grants from NIH-P01-CA-77839 (S. K. D).

Citation Format: Takiko Daikoku, Jumpei Terakawa, Sudhansu K. Dey. A novel mouse model of endometrial cancer: epithelial specific Pten deletion with lactoferrin-iCre. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2289. doi:10.1158/1538-7445.AM2015-2289
Abstract 230: Do-It-Yourself expression microdissection (DIY xMD): A low-cost, high-throughput cell and organelle isolation system
Authors: Tangrea Michael A. , Rosenberg Avi Z. , Fetsch Patricia A. , Armani Michael D. , Xi Liqiang , Raffeld Mark , Pham Tina Thu , Chen Yun , O'Flaherty Neil , Stussman Rebecca , Blackler Adele R. , Du Qiang , Hanson Jeffrey C. , Roth Mark J. , Filie Armando C. , Roh Michael H. , Hipp Jason D. , Emmert-Buck Michael R.
Published: August 2015

Laser-based microdissection technologies precisely recover histological regions or cells from cytology preparations and tissue sections. Yet, due to the high-cost and time-intensiveness of these methods, coarse manual microdissection is the most commonly used tool for laboratory-based studies and clinical applications, enabling rapid procurement but at the expense of reduced cell purity. We present a low-cost, do-it-yourself expression microdissection (DIY xMD) kit that combines the speed and ease-of-use of manual methods with the precision of laser-based instruments. Using DIY xMD, we isolated cell targets including ER+ nuclei and GMS-stained aspergillis fungi from clinical specimens. We also demonstrate the advantage of cellular enrichment in a cytology-based study where BRAF and KRAS mutations were masked by background cells in a manual dissection, yet identifiable after cells were recovered by DIY xMD. The new system makes high-throughput, efficient microdissection accessible to investigators and clinicians throughout the world.

Citation Format: Michael A. Tangrea, Avi Z. Rosenberg, Patricia A. Fetsch, Michael D. Armani, Liqiang Xi, Mark Raffeld, Tina Thu Pham, Yun Chen, Neil O'Flaherty, Rebecca Stussman, Adele R. Blackler, Qiang Du, Jeffrey C. Hanson, Mark J. Roth, Armando C. Filie, Michael H. Roh, Jason D. Hipp, Michael R. Emmert-Buck. Do-It-Yourself expression microdissection (DIY xMD): A low-cost, high-throughput cell and organelle isolation system. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 230. doi:10.1158/1538-7445.AM2015-230
Abstract 2306: Quantitative assessment of one-side directed in vivo angiogenesis
Authors: Prager Gerald , Unseld Matthias , Breuss Johannes , Pausz Clemens , Schabbauer Gernot , Zielinski Christoph , Uhrin Pavel
Published: August 2015

Angiogenesis, the sprouting of new blood vessels from pre-existing vasculature, is an essential process under physiologic and pathological conditions, and is in focus of therapeutic intervention. Reliable in vivo models to study angiogenesis are a pre-requisite for the characterization of novel therapeutic targets. Several methods like the matrigel plug assay, the chick chorioallantoic membrane assay or the cornea assay have addressed the objective to analyse angiogenesis in vivo, but a precise method to quantify one side-directed angiogenesis is lacking so far. Here, we describe an assay, which is based on the use of small matrigel-filled silicone cylinders that are implanted subcutaneously into mice for a period of up to 15 days. In C57BL/6J mice, the effect of the incubation period as well as the dose of the growth factors VEGF and FGF on angiogenic ingrowth was investigated. We describe the appropriate explantation procedure of the silicon tubes, ensuring the complete recovery of the in-grown vessels. The subsequent ex vivo assessment of neo-angiogenesis was then achieved by anti-CD31 as well as anti-endomucin immuno-histochemical staining of ingrown endothelial cells. Additionally, we show that vessel perfusion can be assessed by intravenous injection of a cell staining dye. Altogether, the elaborated protocol offers an effective and reliable assay which facilitates in vivo research on angiogenesis thus might support the development of novel therapeutic agents.

Citation Format: Gerald Prager, Matthias Unseld, Johannes Breuss, Clemens Pausz, Gernot Schabbauer, Christoph Zielinski, Pavel Uhrin. Quantitative assessment of one-side directed in vivo angiogenesis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2306. doi:10.1158/1538-7445.AM2015-2306
Abstract 2313: Characterizing intratumoral cellular heterogeneity in human glioblastoma xenografts by 30+ parameter single-cell mass cytometry
Authors: Simonds Erin F. , Nolan Garry P. , Weiss William A.
Published: August 2015

Introduction: Glioblastoma tumors contain dramatic intratumoral heterogeneity at the level of cellular phenotypes. High-parameter single-cell platforms such as mass cytometry offer an opportunity to simultaneously measure tumor subpopulations with unexpected combinations of differentiation markers, as well as stromal cells, vasculature, and infiltrating immune cells.

Methods: A subcutaneous patient-derived xenograft from an adult with glioblastoma was enzymatically digested into a single-cell suspension. Half of the cells were treated with epidermal growth factor (EGF) at 20 ng/mL for 15 minutes. Both samples were then stained with a 32-marker antibody panel and measured by single-cell mass cytometry. The antibody panel included surface-expressed markers of stemness and differentiation, as well as intracellular markers to monitor cell cycle, apoptosis, and growth factor signaling. Unsupervised spectral clustering was used to segregate >200,000 tumor cells into 13 tumor cell subsets. Inducible signaling phenotypes were quantified in these subsets by comparing binding of phospho-specific antibodies in the stimulated sample, relative to the unstimulated control.

Results: Many previously reported markers of cancer stem cells were detectable, but they did not agree on a single population of cells. Although the CD133+ fraction was rare (<5% of cells), it contained three distinct subpopulations of phenotypically distinct cells. The most informative markers for discriminating the tumor subsets were markers of stemness (i.e. CD133, CD44, Sox2) or integrin expression (i.e. CD49e, CD49f, CD51). The primitive cells (i.e. Sox2+) exhibited higher basal kinase activity across multiple pathways, including markers of cell division (i.e. Histone H3 pSer28). Short-term stimulation with EGF induced distinct responses in each of the different CD133+ subsets.

Conclusions: Single-cell phenotyping by mass cytometry produced rich developmental and functional profiles of glioblastoma cell subsets. Subpopulations of primitive (CD133+) cells can exist in a single glioblastoma tumor and exhibit distinct signaling responses to external stimulation. Altogether, these results validate the use of this platform for investigation of the intratumoral cellular heterogeneity of solid neural tumors with this platform. Work is underway to identify correlations between intratumoral cellular phenotypes and upstream driver mutations in a larger cohort.

Citation Format: Erin F. Simonds, Garry P. Nolan, William A. Weiss. Characterizing intratumoral cellular heterogeneity in human glioblastoma xenografts by 30+ parameter single-cell mass cytometry. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2313. doi:10.1158/1538-7445.AM2015-2313