A comprehensive molecular and clinical analysis of the piRNA pathway genes in ovarian cancer

Ovarian cancer (OC) is one of the most lethal gynecological malignancies, yet molecular mechanisms underlying its origin and progression remain poorly understood. With increasing reports of piRNA pathway deregulation in various cancers, we aimed to better understand its role in OC through a comprehensive analysis of key genes: PIWIL1-4, DDX4, HENMT1, MAEL, PLD6, TDRD1,9 and mutants of PIWIL1 (P1∆17) and PIWIL2 (PL2L60). High-throughput qRT-PCR (n = 45) and CSIOVDB (n = 3431) showed differential gene expression when comparing benign ovarian tumors, low grade OC and high grade serous OC (HGSOC). Significant correlation of disparate piRNA pathway gene expression levels with better progression free, post-progression free and overall survival suggests a complex role of this pathway in OC. We discovered PIWIL3 expression in chemosensitive but not chemoresistant primary HGSOC cells, providing a potential target against chemoresistant disease. As a first, we revealed that follicle stimulating hormone increased PIWIL2 expression in OV-90 cells. PIWIL1, P1∆17, PIWIL2, PL2L60 and MAEL overexpression in vitro and in vivo decreased motility and invasion of OVCAR-3 and OV-90 cells. Interestingly, P1∆17 and PL2L60, induced increased motility and invasion compared to PIWIL1 and PIWIL2. Our results in HGSOC highlight the intricate role piRNA pathway genes play in the development of malignant neoplasms.Eunice Lee, Noor A. Lokman, Martin K. Oehler, Carmela Ricciardelli and Frank Grutzne

Anti-tumour effects of all-trans retinoid acid on serous ovarian cancer

BACKGROUND:Annexin A2 is increased in serous ovarian cancer and plays an essential role in ovarian cancer invasion and metastasis. In combination with S100A10, annexin A2 plays an important role in the plasminogen activator system regulating plasmin production. The aim of this study was to investigate the potential utility of all-trans retinoid acid (ATRA), an inhibitor of the annexin A2-S100A10 signalling pathway, as a new therapeutic against serous ovarian cancer. METHODS:In this study we determined the effects of ATRA treatment (1-5 μM) on annexin A2 and S100A10 expression, plasmin activation, and the ability of ATRA to inhibit serous ovarian cancer cell survival, motility and invasion in vitro. We also employed an ex vivo tissue explant assay to assess response to ATRA treatment in serous ovarian cancers. Cryopreserved serous ovarian cancer tissues were cultured on gelatin sponges for 72 h with ATRA (1 μM). Effects on apoptosis and proliferation were assessed by immunohistochemistry using antibodies to cleaved caspase 3 or Ki67, respectively. RESULTS:Survival of serous ovarian cancer cells (OVCAR-3, OV-90, & OAW28) was significantly decreased by ATRA treatment (1-5 μM). ATRA (1 μM) also significantly decreased proliferation (Ki67 positivity, p = 0.0034), S100A10 protein levels (p = 0.0273), and increased cell apoptosis (cleaved caspase-3 positivity, p = 0.0024) in serous ovarian cancer tissues using the ex vivo tissue explant assay. In OAW28 cells, reduced cell survival following ATRA treatment was associated with a reduction of S100A10 mRNA and protein levels, S100A10 and annexin A2 membrane localization, plasmin generation, motility and invasion. In contrast, ATRA inhibited OV-90 cell survival and invasion but did not affect plasmin activation or S100A10 and annexin A2 expression or membrane localization. CONCLUSIONS:These findings suggest that ATRA inhibits serous ovarian cancer proliferation and invasion via both S100A10 dependant and S100A10 independent mechanisms. Our results show that ATRA has promising potential as a novel therapy against serous ovarian cancer that warrants further evaluation.Noor A. Lokman, Rachel Ho Kavyadharshini Gunasegaran, Wendy M. Bonner, Martin K. Oehler and Carmela Ricciardell

Reduced gonadotrophin receptor expression is associated with a more aggressive ovarian cancer phenotype

Follicle-stimulating hormone (FSH) and luteinising hormone (LH) play important roles in regulating cell growth and proliferation in the ovary. However, few studies have explored the expression of FSH and LH receptors (FSHR and LHCGR) in ovarian cancer, and their functional roles in cancer progression remain inconclusive. This study investigated the potential impact of both mRNA (FSHR, LHCGR) and protein (FSHR, LHCGR) expression on ovarian cancer progression using publicly available online databases, qRT-PCR (high grade serous ovarian cancers, HGSOC, n = 29 and benign ovarian tumors, n = 17) and immunohistochemistry (HGSOC, n = 144). In addition, we investigated the effect of FSHR and LHCGR siRNA knockdown on the pro-metastatic behavior of serous ovarian cancer cells in vitro. High FSHR or high LHCGR expression in patients with all subtypes of high-grade ovarian cancer was significantly associated with longer progression-free survival (PFS) and overall survival (OS). High FSHR protein expression was associated with increased PFS (p = 0.050) and OS (p = 0.025). HGSOC patients with both high FSHR and high LHCGR protein levels had the best survival outcome, whilst both low FSHR and low LHCGR expression was associated with poorest survival (p = 0.019). Knockdown of FSHR significantly increased the invasion of serous ovarian cancer cells (OVCAR3 and COV362) in vitro. LHCGR knockdown also promoted invasion of COV362 cells. This study highlights that lower FSHR and LHCGR expression is associated with a more aggressive epithelial ovarian cancer phenotype and promotes pro-metastatic behaviour.Janelle Cheung, Noor A. Lokman, Riya D. Abraham, Anne M. Macpherson, Eunice Lee , Frank Grutzner, Nicolae Ghinea, Martin K. Oehler and Carmela Ricciardell

Production of renewable diesel from Jatropha curcas oil via pyrolytic-deoxygenation over various multi-wall carbon nanotube-based catalysts

Jatropha curcas is a highly toxic plant that produces seed containing viscous oil with productivity (2 ton/ha), it grows in tropical and sub-tropical regions and offer greater adaptability to a wide range of climatic and soil conditions. Its oils have been noted as an important alternative to produce green diesel via deoxygenation reaction. This study, deoxygenation of jatropha curcas oil (JCO) was carried out over NiO–Fe2O3 and NiO–ZnO catalysts that supported onto multi-walled carbon nanotube (MWCNT). It had found that high Fe and Zn dosages were ineffective in deoxygenation and greatest activity was observed on NiO(20) Fe2O3(5)/MWCNT catalyst. Structure-activity correlations revealed that low metal loading, large density of weak + medium acidic sites and strong basic sites play key role in enhancing the catalytic activities and n-(C15+C17) selectivity. Comparing carbon nanostructures and carbon micron size supported NiO-Fe2O3 revealed that green diesel obtained from NiO–Fe2O3/MWCNT catalysed deoxygenation had the highest heating value and the lowest amounts of oxygen content. Thereby, it confirmed the importance of carbon nanostructure as the catalyst support in improving the diesel quality. Considering the high reusability of NiO-Fe2O3/MWCNT (6 consecutive runs) and superior green diesel properties (flash point, cloud properties and cetane index) demonstrated the NiO–Fe2O3/MWCNT catalyst offers great option in producing excellent properties of green diesel for energy sector

Optical fibre-enabled photoswitching for localised activation of an anti-cancer therapeutic drug

Local activation of an anti-cancer drug when and where needed can improve selectivity and reduce undesirable side effects. Photoswitchable drugs can be selectively switched between active and inactive states by illumination with light; however, the clinical development of these drugs has been restricted by the difficulty in delivering light deep into tissue where needed. Optical fibres have great potential for light delivery in vivo, but their use in facilitating photoswitching in anti-cancer compounds has not yet been explored. In this paper, a photoswitchable chemotherapeutic is switched using an optical fibre, and the cytotoxicity of each state is measured against HCT-116 colorectal cancer cells. The performance of optical-fibre-enabled photoswitching is characterised through its dose response. The UV–Vis spectra confirm light delivered by an optical fibre effectively enables photoswitching. The activated drug is shown to be twice as effective as the inactive drug in causing cancer cell death, characterised using an MTT assay and fluorescent microscopy. This is the first study in which a photoswitchable anti-cancer compound is switched using an optical fibre and demonstrates the feasibility of using optical fibres to activate photoswitchable drugs for potential future clinical applications.Kathryn A. Palasis, Noor A. Lokman, Bryden C. Quirk, Alaknanda Adwal, Loretta Scolaro, Weikun Huang, Carmela Ricciardelli, Martin K. Oehler, Robert A. McLaughlin and Andrew D. Abel

Chemoresistant Cancer Cell Lines Are Characterized by Migratory, Amino Acid Metabolism, Protein Catabolism and IFN1 Signalling Perturbations

Chemoresistance remains the major barrier to effective ovarian cancer treatment. The molecular features and associated biological functions of this phenotype remain poorly understood. We developed carboplatin-resistant cell line models using OVCAR5 and CaOV3 cell lines with the aim of identifying chemoresistance-specific molecular features. Chemotaxis and CAM invasion assays revealed enhanced migratory and invasive potential in OVCAR5-resistant, compared to parental cell lines. Mass spectrometry analysis was used to analyse the metabolome and proteome of these cell lines, and was able to separate these populations based on their molecular features. It revealed signalling and metabolic perturbations in the chemoresistant cell lines. A comparison with the proteome of patient-derived primary ovarian cancer cells grown in culture showed a shared dysregulation of cytokine and type 1 interferon signalling, potentially revealing a common molecular feature of chemoresistance. A comprehensive analysis of a larger patient cohort, including advanced in vitro and in vivo models, promises to assist with better understanding the molecular mechanisms of chemoresistance and the associated enhancement of migration and invasion.Mitchell Acland, Noor A. Lokman, Clifford Young, Dovile Anderson, Mark Condina, Chris Desire, Tannith M. Noye, Wanqi Wang, Carmela Ricciardelli, Darren J. Creek, Martin K. Oehler, Peter Hoffmann, and Manuela Klingler-Hoffman

Matrix Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI MSI) for monitoring of drug response in primary cancer spheroids

Malignant ascites is a fluid, which builds up in the abdomen and contains cancer cells in the form of single cells or multicellular clusters called spheroids. Malignant ascites has been observed in patients suffering from ovarian, cervical, gastric, colorectal, pancreatic, endometrial, or primary liver cancer. The spheroids are believed to play a major role in chemo resistance and metastasis of the cancer. To ease the discomfort of patients, malignant ascites (MA) is often drained from the abdomen using a procedure called paracentesis. MA retrieved via this minimal invasive procedure is a great source for cancer spheroids, which can be used for testing chemotherapeutic drugs and drug combinations. Herein, the existing workflow is adapted to make concurrent monitoring of drug accumulation, drug response, and drug metabolites feasible using primary spheroids or spheroids grown without a scaffolding matrix. To achieve this, those spheroids are embedded in matrigel, before fixing them with formalin. This makes it possible to process, store, and ship samples at room temperature. This new approach might be used to choose the best targeted therapy for each patient and thereby facilitate personalized medicine.Parul Mittal, Zoe K Price, Noor A Lokman, Carmela Ricciardelli, Martin K Oehler, Manuela Klingler-Hoffmann, and Peter Hoffman

MicroRNA-194 promotes prostate cancer metastasis by inhibiting SOCS2

Abstract not availableRajdeep Das, Philip A. Gregory, Rayzel C. Fernandes, Iza Denis, Qingqing Wang, Scott L. Townley, Shuang G. Zhao, Adrienne R. Hanson, Marie A. Pickering, Heather K. Armstrong, Noor A. Lokman, Esmaeil Ebrahimie, Elai Davicioni, Robert B. Jenkins, R. Jeffrey Karnes, Ashley E. Ross, Robert B. Den, Eric A. Klein, Kim N. Chi, Hayley S. Ramshaw, Elizabeth D. Williams, Amina Zoubeidi, Gregory J. Goodall, Felix Y. Feng, Lisa M. Butler, Wayne D. Tilley, and Luke A. Selt