Browsing Over 173 Presentations
Q&A
74P - Promoter hypermethylation of Wnt pathway inhibitor SFRP1 gene and its expression levels in human astrocytomas
- Anja Kafka (Zagreb, HR)
- Anja Kafka (Zagreb, HR)
- Valentina Karin (Zagreb, HR)
- Ljiljana Serman (Zagreb, HR)
- Anja Bukovac (Zagreb, HR)
- Niko Njirić (Zagreb, HR)
- Antonia Jakovcevic (Zagreb, HR)
- Nives Pecina-Slaus (Zagreb, HR)
Abstract
Background
Astrocytomas are the most common primary brain tumors that are on the basis of their histology, molecular characteristics and prognosis classified into 4 different malignancy grades. As one of the key oncogenic pathways in human malignancies that has been associated to many human cancers the Wnt signaling pathway has also been implicated in gliomagenesis. In the present study we aimed to identify the status of
Methods
Promoter hypermethylation of critical molecular component of Wnt signaling – SFRP1 was studied in 24 astrocytomas of different malignancy grades by using methylation-specific PCR (MSP). In order to examine the consequence of hypermethylation, the SFRP1 protein was investigated by immunohistochemistry and analyzed by quantitative stereological analysis.
Results
SFRP1 promoter hypermethylation was found in 32% of astrocytomas. Hypermethylation of SFRP1 promoter was progressively rising in higher astrocytoma grades with the highest significant distribution in glioblastoma (P = 0.042). The expression of SFRP1 protein showed 45.8% of cases with weak or lack of expression, 25% with moderate and 29.2% with strong expression levels. Our study showed that cases with methylated SFRP1 promoter expressed significantly less SFRP1 protein than unmethylated ones (P = 0.031). Furthermore, we have shown that the levels of beta-catenin, pathways indicator of oncogenic activity, were elevated with often nuclear localization. Statistical analysis showed that glioblastoma samples with unmethylated SFRP1 promoter had significantly less beta-catenin (P = 0.033). The activation of Wnt signaling was further confirmed by the expression of its transcriptional activators. Strong expression of LEF1 was significantly associated to higher grades (P = 0.006).
Conclusions
Our findings showed that SFRP1 acts as an antagonist in the evolution of astrocytoma which will contribute to better understanding of astrocytoma molecular profile and offer methylation biomarker of progression.
Legal entity responsible for the study
School of Medicine, University of Zagreb
Funding
Croatian Science Foundation (grant number 6625)
Disclosure
All authors have declared no conflicts of interest.
106P - Antiproliferative effect of trastuzumab and nimotuzumab with EGF on breast cancer cells MCF-7
- Viktoriia V. Nikulina (Kiev, UA)
- Viktoriia V. Nikulina (Kiev, UA)
- L. V. Garmanchuk (Kiev, UA)
- Tetiana V. Nikolaienko (Kiev, UA)
Abstract
Background
In this study, we addressed antiproliferative effects of trastuzumab (Herceptin®) and nimotuzumab (Theraloc®) in a combined application with EGF on MCF-7 cells. The epidermal growth factor (EGF) receptors have been found to be implicated in the ontology and maintenance of tumor tissues, which has fostered the discovery and development of molecularly targeted anti-EGF-receptor therapies. However, the accessibility of chemotherapeutic preparations and humanized antibodies to tumor cells in the G0 phase of the cell cycle is lower than to the cells of theproliferative pool. Given this, antitumor efficiency can potentially be enhanced by synchronizing cells in the proliferative pool. To explore this opportunity, we added EGF to tumor cells in combination with nimotuzumab and trastuzumab (antibodies against EGF-R typeI and II, respectively).
Methods
We added aforementioned compounds to MCF-7 cells in the exponential and stationary phases of growth. Cytotoxic/cytostatic effects and cell survival were assessed by the MTT assay, cytofluorimetry and counting cells stained with trypan blue.
Results
The cytotoxic/cytostatic effect of nimotuzumab in combination with EGF was 56% relative to control, whereas nimotuzumab alone resulted in 45%. In the stationary phase of growth, application of nimotuzumab+EGF did not reveal any effects. Trastuzumab did not produce any antiproliferative effects in combination with EGF during exponential growth, but this combination led to a 60% cytotoxic/cytostatic effect in the stationary phase. Application of trastuzumab alone resulted in a 17% effect during exponential growth and 23% in the stationary phase of MCF-7 cells growth. Trastuzumab in combination with EGF can inhibit cell migratory activity. This combination leads to twice decreased migration area for 2 D growth mode, and 20% for spheroid mode. Trastuzumab also promotes cells attachment to substrate under the serum starvation conditions. Index of adhesion increases in 1,5 times. The combination of trastuzumab and EGF usage promote twice increased adhesion. Nimotuzumab in combination with EGF doesn’t demonstrate any detectible influence on MCF-7 cells adhesion or migration. All the data were statistically significant at p < 0.05.
Conclusions
Utilization humanized antibodies against the EGF receptor in complex with EGF may be considered as a potentially efficient antitumor combination.
Legal entity responsible for the study
Taras Shevchenko National University of Kyiv
Funding
Has not received any funding
Disclosure
All authors have declared no conflicts of interest.
138P - Preclinical testing of NEDD8 and proteasome inhbitors for a treatment-refractory, metastatic high-grade mucinous colorectal cancer patient
- Erica Torchiaro (Candiolo, IT)
- Erica Torchiaro (Candiolo, IT)
- Consalvo Petti (Torino, IT)
- Claudio Isella (Torino, IT)
- Giorgio Corti (Candiolo, IT)
- Monica Montone (Candiolo, IT)
- Benedetta Mussolin (Candiolo, IT)
- Alberto Bardelli (Candiolo, IT)
- Enzo Medico (Torino, IT)
Abstract
Background
Colorectal cancer (CRC) is the third leading cause of death in the world. CRC shows variable phenotypic make-ups; among them, a particularly aggressive histological subtype is the “high grade mucinous” (HGM) adenocarcinoma, highly mucinous, prone to metastasis and typically refractory to treatments. In some cases, HGM is accompanied by a peculiar “signet ring” phenotype of cancer cells. Early stage diagnosis of signet ring HGM is rare, clinical symptoms occur late and most cases are detected at an advanced stage, with a poor overall survival. We demonstrated that a transcriptional signature of HGM displays negative prognosis and sensitivity to the NEDD-8 inhibitor pevonedistat, suggesting the involvement of neddylation- and ubiquitination-based mechanisms in these cases.
Methods
To assess the clinical potential of HGM identification and targeting by pevonedistat or other inhibitor of proteasome pathway, we recently propagated cells and PDXs from a case of early onset, metastatic CRC in a Lynch syndrome patient, refractory to standard care (FOLFOX6, FOLFIRI-Panitumumab) and, surprisingly, also to nivolumab. The tumor was highly mucinous, with signet ring undifferentiated cells.
Results
Mutational analysis on tumor tissue highlighted PIK3CA H1047R mutation and no mutations in KRAS, NRAS or BRAF. Surprisingly, exome analysis on two lesions from a subsequent surgery displayed a different scenario: KRAS G13D but not PIK3CA mutation. Probably these discording results suggest a strong tumor heterogeneity and evolution. Considering the failure of all previous therapies, the lack of actionable genetic alterations and the peculiarity of the phenotypic features, PDX models (organoids and 2D cell cultures) were derived from the two latter lesions and tested for sensitivity to the NEDD8 pathway inhibitor pevonedistat and the proteasome inhibitor bortezomib. Interestingly, all models showed a strong sensitivity to both drugs, in particular to bortezomib.
Conclusions
This is an example of a rare case of HGM colorectal cancer where the integration of several approaches proves useful to identify possible therapeutic strategies for a patient without further standard treatment options.
Legal entity responsible for the study
Candiolo Cancer Institute, FPO-IRCCS
Funding
AIRC, FPRC- Ministero della Salute 5X1000 2011
Disclosure
All authors have declared no conflicts of interest.
51P - Active immunotherapy with a VEGF targeted vaccine HeberSaVax: The road so far and the future ahead
- Mónica Bequet-Romero (Havana, CU)
- Mónica Bequet-Romero (Havana, CU)
- Yanelys M. Díaz (Cubanacán, Playa, CU)
- Javier S. Ramírez (Cubanacán, Playa, CU)
- Katty-Hind Selman-Housein (Havana, CU)
- Francisco Hernández-Bernal (Cubanacán, Playa, CU)
- Ana De la Torre Santos (Santa Clara, CU)
- Jesús Piñero (Santa Clara, CU)
- Cimara Bermúdez (Cubanacán, Playa, CU)
- Jorge V. Gavilondo Cowley (Cubanacán, Playa, CU)
- Marta Ayala Avila (Cubanacán, Playa, CU)
Abstract
Background
The vascular endothelial growth factor (VEGF) plays a central role in angiogenesis and immunosuppressive cascades inherent to tumor development. Success of drugs targeting this growth factor and their receptors signaling cascade built upon these facts. Such passive targeting of VEGF/VEGFR pathway had two major caveats: non-manageable side effects and the induction of resistance phenomena. To overcome some of these problems we design an active immunization approach based on the use of a functionally deficient VEGF 121 isoform as antigen. Herein we present preclinical and clinical development data of HeberSaVax vaccine (formerly known as CIGB-247).
Methods
Safety, antitumoral and immunological effects of HeberSaVax administration were analyzed in mouse, rats, rabbits, non-human primates and in two Phase I clinical trials. Formulations containing the adjuvants VSSP (Very Small Size Particles from Neisseria Meningitides) or Alum phosphate were used. Vaccine was administered once a week (VSSP) or bi-weekly (Alum Phosphate) for 8 weeks. Immune response was evaluated using direct and indirect ELISA methods and IFN-gamma ELISPOT.
Results
HeberSaVax administration inhibits implantation and growth of tumors and metastases favoring a significant increase in animals’ survival. In mice, rats, rabbits and on non-human primates the vaccine administration result in non-toxic induction of sustained antibody titers that specifically neutralized VEGF binding to VEGF receptors. Immunizations also induce VEGF specific cell mediated cytotoxicity. A total of 80 patients were included in the phase I/Ib clinical trials. Results indicated: a good safety profile that allows combining the vaccine with the existing arsenal for cancer therapy; the induction of specific humoral and cellular responses despite the advance stage of patient’s tumors and; an unexpected long term immune responses after 4 years of continuous treatment. As part of the trials three complete responses, one partial response and four stable diseases have been documented.
Conclusions
Altogether these results encourage further development of HeberSaVax vaccine for cancer therapy. Henceforth, a total of four Phase II/III clinical trials are planned to start in 2018 in specific localizations.
Clinical trial identification
RPCEC00000102 and RPCEC00000155
Legal entity responsible for the study
Center for Genetic Engineering and Biotechnology (CIGB)
Funding
Heber Biotec, and Chemo
Disclosure
All authors have declared no conflicts of interest.
26IN - Biological outcomes of CDK4/6 inhibition: Induction of senescence
- Andrew Koff (New York, US)
- Andrew Koff (New York, US)
- Mary E. Klein (New York, US)
- Mark A. Dickson (New York, US)
- Cristina Antonescu (New York, US)
- Li-Xuan Qin (New York, US)
- Scott J. Dooley (Philadelphia, US)
- Gary Schwartz (New York, US)
- Aimee M. Crago (New York, US)
- Samuel Singer (New York, US)
- William D. Tap (New York, US)
Abstract
Background
CDK4/6 inhibitors are being used to treat a variety of human malignancies. In well-differentiated (WDLS) and dedifferentiated (DDLS) liposarcoma we had previously suggested that their clinical promise might be associated with their ability to down-regulate the MDM2 protein. In cultured cell lines, the down-regulation of MDM2, after cells left the cell cycle following CDK4 inhibition, induces them to progress from quiescence into senescence. Today, we will present further evidence supporting the idea that senescence after growth arrest is a mechanism that can account for the activity of CDK4 inhibitors, at least in WD/DDLS.
Results
We have found that PDLIM7 physically associates with MDM2 and prevents MDM2 turnover in quiescent cells induced by treatment with palbociclib (PD0332991). However, if PDLIM7 is sequestered by an intracellular cadherin, CDH18, MDM2 turnover occurs following treatment with PD0332991, driving the quiescent cell into the senescent state. CRISPR knockout of CDH18 can prevent both MDM2 degradation and senescence in cells challenged with CDK4 inhibitor. Establishing the clinical relevancy of this pathway, CDH18 expression is associated with clinical outcome and histologic subtype in patients with advanced WDLS and DDLS from our previous phase II trials. 92% of WDLS and 57% of DDLS had high CDH18 expression judged by immunohistochemistry. High expression of CDH18 in DDLS was associated with improved clinical outcomes.
Conclusions
This supports the hypothesis that the transition from quiescence to senescence has clinical relevance as a mechanism for this class of drugs.
Legal entity responsible for the study
Memorial Sloan Kettering Cancer Center
Funding
National Cancer Institute, USA
Disclosure
All authors have declared no conflicts of interest.
69P - RAB25 a potential therapeutic target in luminal B breast cancer molecular subtype
- LYNDA Addou-Klouche (Marseille, FR)
- LYNDA Addou-Klouche (Marseille, FR)
Abstract
Background
Breast cancer is a complex and heterogeneous disease. Luminal B breast cancers molecular subtype are estrogen receptor-positive (ER+), highly proliferative, resistant to standard therapies and have a poor prognosis. Although they express hormone receptors, their metastatic risk and resistance to hormone therapy and to conventional chemotherapy demand to develop appropriate therapies. Studies have suggested that RAB 25 (Ras-related protein 25) a member of Rab small GTPase family, is involved in Luminal B breast cancer pathogenesis. To better understand this subtype we compared RAB25 DNA copy number aberrations (CNAs) and expression level in luminal B tumors with those observed in breast cancers of the other molecular subtypes.
Methods
To further define molecular alterations associated with the luminal B subtype we studied, with high-Troughput molecular analysis (CGHarray), copy number aberrations and RAB25 gene expression (DNA microarray) deregulation in primary breast cancer samples.
Results
RAB25 gene is not targeted by genomic alterations in a high proportion of breast cancers. RAB25 mRNA expression and RAB 25 protein level was deregulated in breast cancers. Comparison with clinical features between breast tumors with and without RAB 25 deregulation showed that RAB25 high expression was associated with the expression of estrogen receptor (ER), high expression of Ki67 proliferative index and luminal B breast cancer molecular subtypes.
Conclusions
we have identified RAB25 mRNA and protein expression deregulated in luminal B breast cancer molecular subtype suggesting RAB25 as a potential therapeutic target in this aggressive subtype and for which no targeted therapy exists at yet. A better understanding of the involvement of RAB25 in luminal B breast cancer might explain the role in the development and/or hormone resistance of this aggressive subtype.
Legal entity responsible for the study
IPC-Molecular Oncology
Funding
Has not received any funding
Disclosure
The author has declared no conflicts of interest.
101P - Delivery of E-cadherin (CDH1) gene and epidermal growth factor receptor (EGFR) siRNA, using carbonate apatite as a promising vehicle in treatment of breast cancer
- Maeirah A. Ashaie (Selangor, MY)
- Maeirah A. Ashaie (Selangor, MY)
- Iekhsan Othman (Selangor, MY)
- Kyi Kyi Tha (Selangor, MY)
- Ezharul H. Chowdhury (Selangor Malaysia, MY)
Abstract
Background
E-cadherin (CDH1), a cell adhesion glycoprotein is widely associated with breast cancer where it is involved in tumor growth, invasion and metastasis. Down regulation of its gene is hall mark for epithelial-mesenchymal transition, which is an essential process for tumor metastasis. While various pathways are associated with E-cadherin, one of the prominent signaling pathways is via epidermal growth factor reception (EGFR), which is found to be over expressed. Silencing the over expressed EGFR gene by using a specific short-interfering RNA (siRNA), or inducing the expression of CDH1 gene with the help of a plasmid could improve management of malignant cancers by disturbing cancer cell interactions with adjacent cells and extracellular matrix; however, tumor-targeted delivery of these complexes is a major challenge in cancer therapy. This could be overcome by using pH-sensitive carbonate apatite nanoparticles that can enhance the intracellular delivery and release of the bound DNA or siRNA from endosomes to cytosol, after being transported to the tumor.
Methods
Carbonate apatite nanoparticles were employed to deliver CDH1 plasmid and EGFR siRNA individually as well as in combination into human and murine breast cancer cell lines and also into breast tumors induced in mammary pads of 6-8 weeks old Balb/c mice. While MTT assay was performed for cell viability in vitro, tumor regression study was performed in vivo. For tumor regression, two doses of intravenous (IV) treatments were given. Further, protein expression of treated and untreated in vitro samples was confirmed by Western Blot analysis.
Results
Delivery of carbonate apatite nanoparticles with electrostatically associated plasmid or siRNA was found to reduce the tumor burden in mice. The synergistic effect for combined delivery of plasmid CDH1 and EGFR siRNA led to further reduction in the tumor growth. Western blot analysis confirmed the suppression of downstream signaling pathways following delivery of CDH1 gene and EGFR siRNA.
Conclusions
Simultaneous delivery of plasmid CDH1/EGFR siRNA with carbonate apatite nanoparticles could be a promising option for effectively treating breast cancer.
Legal entity responsible for the study
Monash University Malaysia
Funding
FRGS/1/2016/STG05/MUSM/02/3 granted by Ministry of Higher Education, Malaysia
Disclosure
All authors have declared no conflicts of interest.
133P - The global forecast of prostate cancer drug-treatable populations eligible for targeted anticancer therapies (2017-2027)
- Narendra Parihar (Bangalore, IN)
- Narendra Parihar (Bangalore, IN)
Abstract
Background
Targeted anticancer therapies have been approved by the Food and Drug Administration for use in some men with metastatic castrate-resistant prostate cancer (MCRPC). This study aimed to forecast the first-line MCRPC drug-treatable populations (DTP) by global geographic/economic regions over the period 2017-2027.
Methods
Using country-specific cancer registries, prostate cancer incidence was estimated for 45 countries, representing approximately 90% of world population in 2017. Observed correlations between gross domestic product (GDP), prostate cancer risk, and background survival were used to forecast incidence for lower-income countries under study over the next ten years. Diagnosed incident cases were stratified by Tumor Node Metastasis (TNM) stage and risk categories using National Comprehensive Cancer Network (NCCN) 2013 Guidelines. A GDP-based forecast was applied to TNM stage/NCCN risk at diagnosis based on expected improvements in prostate cancer screening for lower-income countries. Metastatic recurrence-free survival was forecast using a function of historical trends to account for improvements in cancer therapy. First-line MCRPC DTP represents the total number of cases eligible for drug treatment in the first-line MCRPC setting each year. Using estimates of stage- and risk-stratified incident cases, and recurrent cases as described above, annualized estimates of first-line MCRPC DTP were obtained. To estimate incident cases and DTP globally, aggregate estimates for each region were divided by the proportion of countries in that region for which direct estimates were made using the methods described above.
Results
In 2017, there were an estimated 331,000 first-line MCRPC DTP worldwide. The first-line MCRPC DTP worldwide will increase by 30% over the period 2017-2027, with higher growth across lower-income countries (40%), than across high-income countries (14%).
Conclusions
The first-line MCRPC DTP is expected to increase globally, primarily driven by increases in diagnosed incidence in conjunction with growing and aging populations in the lower-income regions.
Legal entity responsible for the study
Decision Resources Group, Burlington, MA, USA
Funding
Has not received any funding
Disclosure
The author has declared no conflicts of interest.
Q&A / Panel Discussion
64P - miRNA 200c sensitize pancreatic cancer stem cells to carbon ion beam irradiation
- Sei Sai (Chiba, JP)
- Sei Sai (Chiba, JP)
- Masao Suzuki (Chiba, JP)
- Eun Ho Kim (Seoul, KR)
Abstract
Background
Increasing evidence shows that microRNAs (miRNA), a family of small non-coding RNAs, play a pivotal role in regulating mRNA translation. Recently, some miRNAs have been shown to be involved in regulating cancer stem cell (CSC) properties. Because CSCs are highly resistant to conventional chemotherapy and radiation therapy, and heavy ion radiotherapy is effective in treating those of chemo-radioresistant cancers, in this study we attempt to explore new molecular mechanisms of CSC targeted therapies by carbon ion beam alone or in combination with a miRNA200c mimic.
Methods
Human pancreatic CSCs (CD44+/ESA+) sorted from hum pancreatic cancer cells PK45 and PANC1 were treated with carbon ion beam, X-ray irradiation alone or in combination with a miRNA200c mimic, followed by cell viability assay, colony and spheroid formation ability assay, caspase 3/7 activity assay, and real-time PCR analysis of apoptosis and autophagy-related gene expression were also performed.
Results
The colony, spheroid formation assays confirmed that a subpopulation of CD44+/ESA+ have exactly CSC properties compared with CD44-/ESA- cells in pancreatic cancer cells. Carbon ion beam combined with a miRNA200c mimic significantly decreased CSC viability. The colony, spheroid formation ability of CD44+/ESA+ cells was significantly inhibited by carbon ion beam combined with miRNA200c compared with X-ray irradiation, carbon ion beam alone. Apoptosis analysis showed that caspase activity of 3/7 was significantly enhanced by carbon ion beam in combination with a miRNA200c mimic. Real-time PCR analysis showed that some of apoptosis-related and autophagy-related genes were significantly induced by carbon ion beam alone or in combination with a miRNA200c mimic in pancreatic cancer cells.
Conclusions
Taken together, because the carbon ion beams have a well-defined range with well-localized energy called “spread out Bragg peak (SOBP)”, and release enormous energy at the end of their range, carbon ion beams therefore can kill radioresistant CSCs more than photon beams, and combination with a miRNA200c mimic further enhances those actions, based on the present data.
Legal entity responsible for the study
National Institutes for Quantum and Radiological Science and Technology
Disclosure
All authors have declared no conflicts of interest.