29 results on '"Sharbeen, George"'
Search Results
2. Targeting the undruggable in pancreatic cancer using nano-based gene silencing drugs
- Author
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Kokkinos, John, Ignacio, Rosa Mistica C., Sharbeen, George, Boyer, Cyrille, Gonzales-Aloy, Estrella, Goldstein, David, Australian Pancreatic Cancer Genome Initiative (APGI), McCarroll, Joshua A., and Phillips, Phoebe A.
- Published
- 2020
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3. Ex vivo culture of intact human patient derived pancreatic tumour tissue
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Kokkinos, John, Sharbeen, George, Haghighi, Koroush S., Ignacio, Rosa Mistica C., Kopecky, Chantal, Gonzales-Aloy, Estrella, Youkhana, Janet, Timpson, Paul, Pereira, Brooke A., Ritchie, Shona, Pandzic, Elvis, Boyer, Cyrille, Davis, Thomas P., Butler, Lisa M., Goldstein, David, McCarroll, Joshua A., and Phillips, Phoebe A.
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- 2021
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4. Development of a long term, ex vivo, patient-derived explant model of endometrial cancer.
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van der Woude, Hannah, Phan, Khoi, Kenwright, Diane N., Goossens, Louise, Hally, Kathryn Elizabeth, Currie, Margaret Jane, Kokkinos, John, Sharbeen, George, Phillips, Phoebe A., and Henry, Claire Elizabeth
- Subjects
LEVONORGESTREL ,ENDOMETRIAL cancer ,BODY mass index - Abstract
Incidence of endometrial cancer (EC) is rising in the developed world. The current standard of care, hysterectomy, is often infeasible for younger patients and those with high body mass index. There are limited non-surgical treatment options and a lack of biologically relevant research models to investigate novel alternatives to surgery for EC. The aim of the present study was to develop a long-term, patient-derived explant (PDE) model of early-stage EC and demonstrate its use for investigating predictive biomarkers for a current non-surgical treatment option, the levonorgestrel intra-uterine system (LNG-IUS). Fresh tumour specimens were obtained from patients with early-stage endometrioid EC. Tumours were cut into explants, cultured on media-soaked gelatin sponges for up to 21 days and treated with LNG. Formalin-fixed, paraffin embedded (FFPE) blocks were generated for each explant after 21 days in culture. Tumour architecture and integrity were assessed by haematoxylin and eosin (H&E) and immunohistochemistry (IHC). IHC was additionally performed for the expression of five candidate biomarkers of LNG resistance. The developed ex vivo PDE model is capable of culturing explants from early-stage EC tumours long-term (21 Days). This model can complement existing models and may serve as a tool to validate results obtained in higher-throughput in vitro studies. Our study provides the foundation to validate the extent to which EC PDEs reflect patient response in future research. [ABSTRACT FROM AUTHOR]
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- 2024
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5. Novel therapeutics and preclinical imaging for pancreatic cancer - view from the lab
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Akerman, Anouschka, Vennin, Claire, Sharbeen, George, Warren, Sean C, Timpson, Paul, and Phillips, Phoebe A
- Published
- 2016
6. Incorporation of dUTP does not mediate mutation of A: T base pairs in Ig genes in vivo
- Author
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Sharbeen, George, Cook, Adam J. L., Lau, K. K. Edwin, Raftery, Joanna, Yee, Christine W. Y., and Jolly, Christopher J.
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- 2010
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7. How to exploit different endocytosis pathways to allow selective delivery of anticancer drugs to cancer cells over healthy cells.
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Cong, Vu Thanh, Tilley, Richard D., Sharbeen, George, Phillips, Phoebe A., Gaus, Katharina, and Gooding, J. Justin
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- 2021
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8. ROR1 and ROR2 expression in pancreatic cancer.
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Liu, Dongli, Sharbeen, George, Phillips, Phoebe, Australian Pancreatic Cancer Genome Initiative, Johns, Amber L., Gill, Anthony J., Chantrill, Lorraine A., Timpson, Paul, Chou, Angela, Pajic, Marina, Dwarte, Tanya, Herrmann, David, Vennin, Claire, Cox, Thomas R., Pereira, Brooke, Ritchie, Shona, Reed, Daniel A., Chambers, Cecilia R., Metcalf, Xanthe, and Nobis, Max
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GENE expression , *PANCREATIC cancer , *PROGNOSIS , *OVERALL survival , *DRUG target - Abstract
Background: The Wnt receptors ROR1 and ROR2 are generating increased interest as cancer therapeutic targets but remain understudied in pancreatic ductal adenocarcinoma (PDAC). Compared to canonical Wnt/ β-catenin signalling, the role of noncanonical Wnt signalling in PDAC remains largely unknown. Only one study has investigated the prognostic significance of the noncanonical Wnt signalling receptor, ROR2 in PDAC. No studies have investigated the prognostic role of ROR1 in PDAC.Methods: Here, we performed analysis of ROR1 and ROR2 mRNA expression in three publicly available datasets ICGC-PACA-AU (n = 81), TCGA-PAAD (n = 150) and CPTAC-PDAC (n = 137). ROR1 and ROR2 protein expression from the CPTAC-PDAC discovery cohort were also analysed. Immunohistochemistry (IHC) using the validated anti ROR1 monoclonal antibody (4A5) was performed on the Australian Pancreatic Cancer Genome Initiative (APGI) cohort of PDAC samples (n = 152). Association between ROR1 cytoplasmic staining intensity and clinicopathological parameters including stage, grade and overall survival (OS) was investigated.Results: High ROR1 mRNA expression levels correlated with a favourable OS outcome in all of the ICGC-PACA-AU, TCGA-PAAD and CPTAC-PDAC cohorts. ROR1 protein expression was not associated with stage, grade or OS in the APGI cohort.Conclusion: ROR1 and ROR2 have potential as prognostic markers when measured at the mRNA level in PDAC. Our IHC cohort did not support ROR1 protein expression in predicting OS, and highlighted the discrepancy of prognostic biomarkers when measured by MS, IHC and RNAseq. [ABSTRACT FROM AUTHOR]- Published
- 2021
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9. Can the Shape of Nanoparticles Enable the Targeting to Cancer Cells over Healthy Cells?
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Cong, Vu Thanh, Wang, Wenqian, Tilley, Richard D., Sharbeen, George, Phillips, Phoebe A., Gaus, Katharina, and Gooding, John Justin
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BREAST cancer ,NANOPARTICLES ,PANCREATIC cancer ,NANOPARTICLE size ,CANCER cells ,EPITHELIAL cells - Abstract
Macropinocytosis is a consequence of oncogenic alterations of cancer cells while most healthy cells are non‐macropinocytic. It is currently unclear whether macropinocytic cancer cells can be targeted rather than healthy cells, by adjusting the shape and size of nanoparticles. Herein, the endocytosis of two differently shaped nanoparticles; nanorods and nanospheres are compared in cancer and healthy cells. The cells are breast epithelial cancer cells (MCF7) and breast epithelial healthy cells (MCF10A) and pancreas cancer cells (PANC‐1 cells) and non‐tumourogenic patient‐derived cancer‐associated fibroblasts (CAFs). The endocytosis pathway is quantified by a combination of pair correlation microscopy and endocytosis inhibitors. MCF7 cells use clathrin‐mediated endocytosis and macropinocytosis to take up the nanorods while MCF10A cells use predominantly clathrin‐mediated endocytosis. Based on the comparison of endocytic behavior of cancer and healthy cells, MCF7 cells can be induced to take up more nanorods and suppress the metabolism and endocytosis of nanorods in MCF10A cells. The nanorods allow targeting to breast cancer MCF7 cells and pancreas cancer cells over the healthy cells. This study opens exciting possibilities for shape to target the cancer cells over healthy cells, by adjusting nanoparticle shape. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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10. Biochemical characterization of the multidrug regulator QacR distinguishes residues that are crucial to multidrug binding and induction of qacA transcription
- Author
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Peters, Kate M., Sharbeen, George, Theis, Torsten, Skurray, Ronald A., and Brown, Melissa H.
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DNA-ligand interactions -- Research ,Glutamine -- Chemical properties ,Glutamine -- Structure ,Leucine -- Chemical properties ,Leucine -- Structure ,Staphylococcus aureus -- Research ,Tyrosine -- Chemical properties ,Tyrosine -- Structure ,Biological sciences ,Chemistry - Abstract
The studies on functional significance of hydrophobic, aromatic, and polar residues characteristic of the rhodamine 6G pocket and the proximal [Tyr.sup.92] which facilitated the transcriptionally active conformation are reported. The results highlight the significant role of [Leu.sup.54], [Tyr.sup.93], [Gln.sup.96], and [Tyr.sup.123] residues to QacR-mediated derepression of qacA transcription following ligand binding in the distal subpocket which might be important for the general mechanism irrespective of the bound ligand.
- Published
- 2009
11. Targeting a solute carrier to metabolically reprogram tumour-promoting stromal cells in pancreatic cancer
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Akerman, Anouschka, Sharbeen, George, Holst, Jeff, Youkhana, Janet, McCarroll, Joshua, Goldstein, David, Erkan, Mert, and Phillips, Phoebe
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- 2018
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12. Proximity to AGCT sequences dictates MMR-independent versus MMR-dependent mechanisms for AID-induced mutation via UNG2.
- Author
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Sanchai Thientosapol, Eddy, Sharbeen, George, Lau, K. K. Edwin, Bosnjak, Daniel, Durack, Timothy, Stevanovski, Igor, Weninger, Wolfgang, and Jolly, Christopher J.
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- 2017
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13. MutY-Homolog modulates pancreatic cancer cell survival and chemoresistance
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Sharbeen, George, Youkhana, Janet, McCarroll, Joshua, Akerman, Anouschka, Goldstein, David, and Phillips, Phoebe
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- 2016
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14. A Rationally Optimized Nanoparticle System for the Delivery of RNA Interference Therapeutics into Pancreatic Tumors in Vivo.
- Author
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Teo, Joann, McCarroll, Joshua A., Boyer, Cyrille, Janet Youkhana, Sagnella, Sharon M., Duong, Hien T. T., Jie Liu, Sharbeen, George, Goldstein, David, Davis, Thomas P., Kavallaris, Maria, and Phillips, Phoebe A.
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- 2016
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15. Role of pancreatic stellate cells in chemoresistance in pancreatic cancer.
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McCarroll, Joshua A., Naim, Stephanie, Sharbeen, George, Russia, Nelson, Lee, Julia, Kavallaris, Maria, Goldstein, David, and Phillips, Phoebe A.
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PANCREATIC cancer ,CANCER cells ,CANCER invasiveness ,CANCER research ,PHYSIOLOGICAL research - Abstract
Pancreatic cancer is highly chemoresistant. A major contributing factor is the characteristic extensive stromal or fibrotic reaction, which comprises up to 90% of the tumor volume. Over the last decade there has been intensive research into the role of the pro-fibrogenic pancreatic stellate cells (PSCs) and their interaction with pancreatic cancer cells. As a result of the significant alterations in the tumor microenvironment following activation of PSCs, tumor progression, and chemoresistance is enhanced. This review will discuss how PSCs contribute to chemoresistance in pancreatic cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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16. Incorporation of dUTP does not mediate mutation of A:T base pairs in Ig genes in vivo.
- Author
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Sharbeen, George, Cook, Adam J. L., Lau, K. K. Edwin, Raftery, Joanna, Yee, Christine W. Y., and Jolly, Christopher J.
- Published
- 2010
- Full Text
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17. Does the Microenvironment Hold the Hidden Key for Functional Precision Medicine in Pancreatic Cancer?
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Kokkinos, John, Jensen, Anya, Sharbeen, George, McCarroll, Joshua A., Goldstein, David, Haghighi, Koroush S., Phillips, Phoebe A., and Putoczki, Tracy
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TISSUE analysis ,THERAPEUTIC use of monoclonal antibodies ,THERAPEUTIC use of antineoplastic agents ,PANCREATIC tumors ,BIOLOGICAL models ,CELL culture ,INDIVIDUALIZED medicine ,CELL physiology ,DUCTAL carcinoma ,CELL lines ,DRUG resistance in cancer cells - Abstract
Simple Summary: Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest of all major cancers. We now know that a 'one-size-fits-all' approach when treating PDAC patients does not work, so research has focused on developing personalised treatment strategies based on the biology of each patient's tumour. However, most of these strategies ignore the presence of the characteristic fibrotic PDAC stroma which can act like a fortress to protect the tumour from chemotherapy and drive its aggressive nature. In this review, we summarise the role of this 'fortress' in protecting PDAC from chemotherapy and highlight the need for pre-clinical models that are informing precision medicine to reflect this hallmark feature of PDAC. We also propose the power of ex vivo whole-tissue culture models that mimic the tumour and its surrounding fortress to inform personalised treatment for PDAC. Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers and no significant improvement in patient survival has been seen in the past three decades. Treatment options are limited and selection of chemotherapy in the clinic is usually based on the performance status of a patient rather than the biology of their disease. In recent years, research has attempted to unlock a personalised treatment strategy by identifying actionable molecular targets in tumour cells or using preclinical models to predict the effectiveness of chemotherapy. However, these approaches rely on the biology of PDAC tumour cells only and ignore the importance of the microenvironment and fibrotic stroma. In this review, we highlight the importance of the microenvironment in driving the chemoresistant nature of PDAC and the need for preclinical models to mimic the complex multi-cellular microenvironment of PDAC in the precision medicine pipeline. We discuss the potential for ex vivo whole-tissue culture models to inform precision medicine and their role in developing novel therapeutic strategies that hit both tumour and stromal compartments in PDAC. Thus, we highlight the critical role of the tumour microenvironment that needs to be addressed before a precision medicine program for PDAC can be implemented. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
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18. Cancer-Associated Fibroblasts in Pancreatic Ductal Adenocarcinoma Determine Response to SLC7A11 Inhibition.
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Sharbeen G, McCarroll JA, Akerman A, Kopecky C, Youkhana J, Kokkinos J, Holst J, Boyer C, Erkan M, Goldstein D, Timpson P, Cox TR, Pereira BA, Chitty JL, Fey SK, Najumudeen AK, Campbell AD, Sansom OJ, Ignacio RMC, Naim S, Liu J, Russia N, Lee J, Chou A, Johns A, Gill AJ, Gonzales-Aloy E, Gebski V, Guan YF, Pajic M, Turner N, Apte MV, Davis TP, Morton JP, Haghighi KS, Kasparian J, McLean BJ, Setargew YF, and Phillips PA
- Subjects
- Amino Acid Transport System y+ genetics, Amino Acid Transport System y+ immunology, Animals, Apoptosis, Cancer-Associated Fibroblasts immunology, Cancer-Associated Fibroblasts pathology, Carcinoma, Pancreatic Ductal genetics, Carcinoma, Pancreatic Ductal metabolism, Carcinoma, Pancreatic Ductal pathology, Cell Proliferation, Female, Humans, Mice, Mice, Inbred BALB C, Mice, Nude, Pancreatic Neoplasms genetics, Pancreatic Neoplasms metabolism, Pancreatic Neoplasms pathology, Prognosis, Survival Rate, Tumor Cells, Cultured, Xenograft Model Antitumor Assays, Amino Acid Transport System y+ antagonists & inhibitors, Antibodies, Monoclonal pharmacology, Cancer-Associated Fibroblasts drug effects, Carcinoma, Pancreatic Ductal prevention & control, Gene Expression Regulation, Neoplastic drug effects, Pancreatic Neoplasms prevention & control, Tumor Microenvironment
- Abstract
Cancer-associated fibroblasts (CAF) are major contributors to pancreatic ductal adenocarcinoma (PDAC) progression through protumor signaling and the generation of fibrosis, the latter of which creates a physical barrier to drugs. CAF inhibition is thus an ideal component of any therapeutic approach for PDAC. SLC7A11 is a cystine transporter that has been identified as a potential therapeutic target in PDAC cells. However, no prior study has evaluated the role of SLC7A11 in PDAC tumor stroma and its prognostic significance. Here we show that high expression of SLC7A11 in human PDAC tumor stroma, but not tumor cells, is independently prognostic of poorer overall survival. Orthogonal approaches showed that PDAC-derived CAFs are highly dependent on SLC7A11 for cystine uptake and glutathione synthesis and that SLC7A11 inhibition significantly decreases CAF proliferation, reduces their resistance to oxidative stress, and inhibits their ability to remodel collagen and support PDAC cell growth. Importantly, specific ablation of SLC7A11 from the tumor compartment of transgenic mouse PDAC tumors did not affect tumor growth, suggesting the stroma can substantially influence PDAC tumor response to SLC7A11 inhibition. In a mouse orthotopic PDAC model utilizing human PDAC cells and CAFs, stable knockdown of SLC7A11 was required in both cell types to reduce tumor growth, metastatic spread, and intratumoral fibrosis, demonstrating the importance of targeting SLC7A11 in both compartments. Finally, treatment with a nanoparticle gene-silencing drug against SLC7A11, developed by our laboratory, reduced PDAC tumor growth, incidence of metastases, CAF activation, and fibrosis in orthotopic PDAC tumors. Overall, these findings identify an important role of SLC7A11 in PDAC-derived CAFs in supporting tumor growth. SIGNIFICANCE: This study demonstrates that SLC7A11 in PDAC stromal cells is important for the tumor-promoting activity of CAFs and validates a clinically translatable nanomedicine for therapeutic SLC7A11 inhibition in PDAC., (©2021 American Association for Cancer Research.)
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- 2021
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19. Interfacial Curvature in Confined Coculture Directs Stromal Cell Activity with Spatial Corralling of Pancreatic Cancer Cells.
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Nemec S, Lam J, Zhong J, Heu C, Timpson P, Li Q, Youkhana J, Sharbeen G, Phillips PA, and Kilian KA
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- Coculture Techniques, Humans, Male, Mechanotransduction, Cellular, Stromal Cells, Tumor Microenvironment, Cancer-Associated Fibroblasts, Pancreatic Neoplasms
- Abstract
Interfacial cues in the tumor microenvironment direct the activity and assembly of multiple cell types. Pancreatic cancer, along with breast and prostate cancers, is enriched with cancer-associated fibroblasts (CAFs) that activate to coordinate the deposition of the extracellular matrix, which can comprise over 90% of the tumor mass. While it is clear that matrix underlies the severity of the disease, the relationship between stromal-tumor cell assembly and cell-matrix dynamics remains elusive. Micropatterned hydrogels deconstruct the interplay between matrix stiffness and geometric confinement, guiding heterotypic cell populations and matrix assembly in pancreatic cancer. Interfacial cues at the perimeter of microislands guide CAF migration and direct cancer cell assembly. Computational modeling shows curvature-stress dependent cellular localization for cancer and CAFs in coculture. Regions of convex curvature enhance edge stress that activates a myofibroblast phenotype in the CAFs with migration and increased collagen I deposition, ultimately leading to a central "corralling" of cancer cells. Inhibiting mechanotransduction pathways decreases CAF activation and the associated corralling phenotype. Together, this work reveals how interfacial biophysical cues underpin aspects of stromal desmoplasia, a hallmark of disease severity and chemoresistance in the pancreatic, breast, and prostate cancers, thereby providing a tool to expand stroma-targeting therapeutic strategies., (© 2021 Wiley-VCH GmbH.)
- Published
- 2021
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20. Identification of Novel Medulloblastoma Cell-Targeting Peptides for Use in Selective Chemotherapy Drug Delivery.
- Author
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Tjandra KC, McCarthy N, Yang L, Laos AJ, Sharbeen G, Phillips PA, Forgham H, Sagnella SM, Whan RM, Kavallaris M, Thordarson P, and McCarroll JA
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- Antineoplastic Agents pharmacokinetics, Antineoplastic Agents pharmacology, Blood-Brain Barrier drug effects, Blood-Brain Barrier metabolism, Brain Neoplasms metabolism, Cell Line, Cell Line, Tumor, Child, Doxorubicin pharmacokinetics, Doxorubicin pharmacology, Drug Carriers chemistry, Drug Delivery Systems, Humans, Medulloblastoma metabolism, Oligopeptides chemistry, Peptide Library, Antineoplastic Agents administration & dosage, Brain Neoplasms drug therapy, Doxorubicin administration & dosage, Drug Carriers metabolism, Medulloblastoma drug therapy, Oligopeptides metabolism
- Abstract
Medulloblastoma is a malignant brain tumor diagnosed in children. Chemotherapy has improved survival rates to approximately 70%; however, children are often left with long-term treatment side effects. New therapies that maintain a high cure rate while reducing off-target toxicity are required. We describe for the first time the use of a bacteriophage-peptide display library to identify heptapeptides that bind to medulloblastoma cells. Two heptapeptides that demonstrated high [E1-3 ( 1 )] or low [E1-7 ( 2 )] medulloblastoma cell binding affinity were synthesized. The potential of the peptides to deliver a therapeutic drug to medulloblastoma cells with specificity was investigated by conjugating E1-3 ( 1 ) or E1-7 ( 2 ) to doxorubicin ( 5 ). Both peptide-drug conjugates were cytotoxic to medulloblastoma cells. E1-3 doxorubicin ( 3 ) could permeabilize an in vitro blood-brain barrier and showed a marked reduction in cytotoxicity compared to free doxorubicin ( 5 ) in nontumor cells. This study provides proof-of-concept for developing peptide-drug conjugates to inhibit medulloblastoma cell growth while minimizing off-target toxicity.
- Published
- 2020
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21. The Use of Star Polymer Nanoparticles for the Delivery of siRNA to Mouse Orthotopic Pancreatic Tumor Models.
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McCarroll JA, Sharbeen G, Kavallaris M, and Phillips PA
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- Animals, Cell Line, Tumor, Cell Proliferation drug effects, Gene Silencing drug effects, Humans, Mice, Nanoparticles therapeutic use, Pancreatic Neoplasms genetics, Polymers chemistry, Polymers pharmacology, RNA Interference drug effects, RNA, Small Interfering chemistry, RNA, Small Interfering pharmacology, Tubulin genetics, Tubulin Modulators chemistry, Tubulin Modulators pharmacology, Xenograft Model Antitumor Assays, Drug Delivery Systems methods, Nanoparticles chemistry, Pancreatic Neoplasms therapy, RNA, Small Interfering genetics
- Abstract
Pancreatic cancer is a lethal malignancy which is refractory to most chemotherapy drugs. Recent landmark studies have shed new light on the complex genetic heterogeneity of pancreatic cancer and provide an opportunity to utilize "precision-based medicines" to target genes based on the genetic profile of an individual's tumor to increase the efficiency of chemotherapy and decrease tumor growth and metastases. Gene-silencing drugs in the form of short-interfering RNA (siRNA) have the potential to play an important role in precision medicine for pancreatic cancer by silencing the expression of genes including those considered difficult to inhibit (undruggable) using chemical agents. However, before siRNA can reach its clinical potential a delivery vehicle is needed to carry siRNA across the cell membrane and into the cytoplasm of the cell. Herein, we detail the methods required to use star polymer nanoparticles to deliver siRNA to pancreatic tumors in an orthotopic pancreatic cancer mouse model to silence the expression of an "undruggable" gene (βIII-tubulin) that regulates pancreatic cancer growth and chemosensitivity.
- Published
- 2019
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22. Proximity to AGCT sequences dictates MMR-independent versus MMR-dependent mechanisms for AID-induced mutation via UNG2.
- Author
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Thientosapol ES, Sharbeen G, Lau KKE, Bosnjak D, Durack T, Stevanovski I, Weninger W, and Jolly CJ
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- Adoptive Transfer, Animals, Base Pairing, Base Sequence, Male, Mice, Inbred C57BL, Uracil metabolism, Uracil-DNA Glycosidase genetics, Cytidine Deaminase metabolism, DNA Mismatch Repair, DNA Repair, Mutation, Uracil-DNA Glycosidase metabolism
- Abstract
AID deaminates C to U in either strand of Ig genes, exclusively producing C:G/G:C to T:A/A:T transition mutations if U is left unrepaired. Error-prone processing by UNG2 or mismatch repair diversifies mutation, predominantly at C:G or A:T base pairs, respectively. Here, we show that transversions at C:G base pairs occur by two distinct processing pathways that are dictated by sequence context. Within and near AGCT mutation hotspots, transversion mutation at C:G was driven by UNG2 without requirement for mismatch repair. Deaminations in AGCT were refractive both to processing by UNG2 and to high-fidelity base excision repair (BER) downstream of UNG2, regardless of mismatch repair activity. We propose that AGCT sequences resist faithful BER because they bind BER-inhibitory protein(s) and/or because hemi-deaminated AGCT motifs innately form a BER-resistant DNA structure. Distal to AGCT sequences, transversions at G were largely co-dependent on UNG2 and mismatch repair. We propose that AGCT-distal transversions are produced when apyrimidinic sites are exposed in mismatch excision patches, because completion of mismatch repair would require bypass of these sites., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2017
- Full Text
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23. MutY-Homolog (MYH) inhibition reduces pancreatic cancer cell growth and increases chemosensitivity.
- Author
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Sharbeen G, Youkhana J, Mawson A, McCarroll J, Nunez A, Biankin A, Johns A, Goldstein D, and Phillips P
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- Animals, Apoptosis drug effects, Carcinoma, Pancreatic Ductal genetics, Carcinoma, Pancreatic Ductal metabolism, Carcinoma, Pancreatic Ductal pathology, Cell Cycle drug effects, Cell Line, Tumor, DNA Glycosylases genetics, Deoxycytidine analogs & derivatives, Deoxycytidine pharmacology, Dose-Response Relationship, Drug, Female, Gene Expression Regulation, Neoplastic, Humans, Mice, Inbred BALB C, Mice, Nude, Oxidative Stress drug effects, Paclitaxel pharmacology, Pancreatic Neoplasms genetics, Pancreatic Neoplasms metabolism, Pancreatic Neoplasms pathology, RNA Interference, Signal Transduction drug effects, Time Factors, Transfection, Tumor Burden drug effects, Tumor Hypoxia, Tumor Microenvironment, Vincristine pharmacology, Xenograft Model Antitumor Assays, Gemcitabine, Antineoplastic Agents pharmacology, Carcinoma, Pancreatic Ductal drug therapy, Cell Proliferation drug effects, DNA Glycosylases metabolism, Pancreatic Neoplasms drug therapy
- Abstract
Patients with pancreatic ductal adenocarcinoma (PC) have a poor prognosis due to metastases and chemoresistance. PC is characterized by extensive fibrosis, which creates a hypoxic microenvironment, and leads to increased chemoresistance and intracellular oxidative stress. Thus, proteins that protect against oxidative stress are potential therapeutic targets for PC. A key protein that maintains genomic integrity against oxidative damage is MutY-Homolog (MYH). No prior studies have investigated the function of MYH in PC cells. Using siRNA, we showed that knockdown of MYH in PC cells 1) reduced PC cell proliferation and increased apoptosis; 2) further decreased PC cell growth in the presence of oxidative stress and chemotherapy agents (gemcitabine, paclitaxel and vincristine); 3) reduced PC cell metastatic potential; and 4) decreased PC tumor growth in a subcutaneous mouse model in vivo. The results from this study suggest MYH may be a novel therapeutic target for PC that could potentially improve patient outcome by reducing PC cell survival, increasing the efficacy of existing drugs and reducing metastatic spread.
- Published
- 2017
- Full Text
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24. Exploiting base excision repair to improve therapeutic approaches for pancreatic cancer.
- Author
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Sharbeen G, McCarroll J, Goldstein D, and Phillips PA
- Abstract
Pancreatic ductal adenocarcinoma (PDA) is a highly chemoresistant and metastatic disease with a dismal 5-year survival rate of 6%. More effective therapeutic targets and approaches are urgently needed to tackle this devastating disease. The base excision repair (BER) pathway has been identified as a predictor of therapeutic response, prognostic factor, and therapeutic target in a variety of cancers. This review will discuss our current understanding of BER in PDA and its potential to improve PDA treatment.
- Published
- 2015
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25. Correction to "effective delivery of siRNA into cancer cells and tumors using well-defined biodegradable cationic star polymers".
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Boyer C, Teo J, Phillips P, Erlich RB, Sagnella S, Sharbeen G, Dwarte T, Duong HT, Goldstein D, Davis TP, Kavallaris M, and McCarroll J
- Published
- 2015
- Full Text
- View/download PDF
26. βIII-tubulin: a novel mediator of chemoresistance and metastases in pancreatic cancer.
- Author
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McCarroll JA, Sharbeen G, Liu J, Youkhana J, Goldstein D, McCarthy N, Limbri LF, Dischl D, Ceyhan GO, Erkan M, Johns AL, Biankin AV, Kavallaris M, and Phillips PA
- Subjects
- Animals, Anoikis genetics, Apoptosis genetics, Blotting, Western, Cell Cycle genetics, Cell Line, Tumor, Cell Proliferation genetics, Gene Expression Regulation, Neoplastic, Humans, Immunohistochemistry, Mice, Inbred BALB C, Mice, Nude, Neoplasm Metastasis, Pancreatic Neoplasms metabolism, Pancreatic Neoplasms pathology, RNAi Therapeutics, Reverse Transcriptase Polymerase Chain Reaction, Tubulin metabolism, Xenograft Model Antitumor Assays, Drug Resistance, Neoplasm genetics, Pancreatic Neoplasms genetics, RNA Interference, Tubulin genetics
- Abstract
Pancreatic cancer is a leading cause of cancer-related deaths in Western societies. This poor prognosis is due to chemotherapeutic drug resistance and metastatic spread. Evidence suggests that microtubule proteins namely, β-tubulins are dysregulated in tumor cells and are involved in regulating chemosensitivity. However, the role of β-tubulins in pancreatic cancer are unknown. We measured the expression of different β-tubulin isotypes in pancreatic adenocarcinoma tissue and pancreatic cancer cells. Next, we used RNAi to silence βIII-tubulin expression in pancreatic cancer cells, and measured cell growth in the absence and presence of chemotherapeutic drugs. Finally, we assessed the role of βIII-tubulin in regulating tumor growth and metastases using an orthotopic pancreatic cancer mouse model. We found that βIII-tubulin is highly expressed in pancreatic adenocarcinoma tissue and pancreatic cancer cells. Further, we demonstrated that silencing βIII-tubulin expression reduced pancreatic cancer cell growth and tumorigenic potential in the absence and presence of chemotherapeutic drugs. Finally, we demonstrated that suppression of βIII-tubulin reduced tumor growth and metastases in vivo. Our novel data demonstrate that βIII-tubulin is a key player in promoting pancreatic cancer growth and survival, and silencing its expression may be a potential therapeutic strategy to increase the long-term survival of pancreatic cancer patients.
- Published
- 2015
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27. Effective delivery of siRNA into cancer cells and tumors using well-defined biodegradable cationic star polymers.
- Author
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Boyer C, Teo J, Phillips P, Erlich RB, Sagnella S, Sharbeen G, Dwarte T, Duong HT, Goldstein D, Davis TP, Kavallaris M, and McCarroll J
- Subjects
- Animals, Blotting, Western, Cell Line, Tumor, Chromatography, Gel, Female, Humans, Magnetic Resonance Spectroscopy, Mice, Mice, Inbred BALB C, Mice, Nude, Microscopy, Electron, Transmission, Nanoparticles chemistry, Reverse Transcriptase Polymerase Chain Reaction, Polymers chemistry, RNA, Small Interfering administration & dosage
- Abstract
Cancer is one of the most common causes of death worldwide. Two types of cancer that have high mortality rates are pancreatic and lung cancer. Despite improvements in treatment strategies, resistance to chemotherapy and the presence of metastases are common. Therefore, novel therapies which target and silence genes involved in regulating these processes are required. Short-interfering RNA (siRNA) holds great promise as a therapeutic to silence disease-causing genes. However, siRNA requires a delivery vehicle to enter the cell to allow it to silence its target gene. Herein, we report on the design and synthesis of cationic star polymers as novel delivery vehicles for siRNA to silence genes in pancreatic and lung cancer cells. Dimethylaminoethyl methacrylate (DMAEMA) was polymerized via reversible addition-fragmentation transfer polymerization (RAFT) and then chain extended in the presence of both cross-linkers N,N-bis(acryloyl)cistamine and DMAEMA, yielding biodegradable well-defined star polymers. The star polymers were characterized by transmission electron microscopy, dynamic light scattering, ζ potential, and gel permeation chromatography. Importantly, the star polymers were able to self-assemble with siRNA and form small uniform nanoparticle complexes. Moreover, the ratios of star polymer required to complex siRNA were nontoxic in both pancreatic and lung cancer cells. Treatment with star polymer-siRNA complexes resulted in uptake of siRNA into both cell lines and a significant decrease in target gene mRNA and protein levels. In addition, delivery of clinically relevant amounts of siRNA complexed to the star polymer were able to silence target gene expression by 50% in an in vivo tumor setting. Collectively, these results provide the first evidence of well-defined small cationic star polymers to deliver active siRNA to both pancreatic and lung cancer cells and may be a valuable tool to inhibit key genes involved in promoting chemotherapy drug resistance and metastases.
- Published
- 2013
- Full Text
- View/download PDF
28. Ectopic restriction of DNA repair reveals that UNG2 excises AID-induced uracils predominantly or exclusively during G1 phase.
- Author
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Sharbeen G, Yee CW, Smith AL, and Jolly CJ
- Subjects
- Animals, Genes, Immunoglobulin physiology, Humans, Mice, Mice, Inbred C57BL, Plasmids genetics, Recombinant Fusion Proteins metabolism, Time-Lapse Imaging, Transduction, Genetic, Uracil Nucleotides metabolism, Cytidine Deaminase metabolism, DNA Glycosylases metabolism, DNA Repair physiology, G1 Phase physiology, Genes, Immunoglobulin genetics, Immunoglobulin Class Switching physiology
- Abstract
Immunoglobulin (Ig) affinity maturation requires the enzyme AID, which converts cytosines (C) in Ig genes into uracils (U). This alone produces C:G to T:A transition mutations. Processing of U:G base pairs via U N-glycosylase 2 (UNG2) or MutSα generates further point mutations, predominantly at G:C or A:T base pairs, respectively, but it is unclear why processing is mutagenic. We aimed to test whether the cell cycle phase of U processing determines fidelity. Accordingly, we ectopically restricted UNG2 activity in vivo to predefined cell cycle phases by fusing a UNG2 inhibitor peptide to cell cycle-regulated degradation motifs. We found that excision of AID-induced U by UNG2 occurs predominantly during G1 phase, inducing faithful repair, mutagenic processing, and class switching. Surprisingly, UNG2 does not appear to process U:G base pairs at all in Ig genes outside G1 phase.
- Published
- 2012
- Full Text
- View/download PDF
29. A novel role of dipeptidyl peptidase 9 in epidermal growth factor signaling.
- Author
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Yao TW, Kim WS, Yu DM, Sharbeen G, McCaughan GW, Choi KY, Xia P, and Gorrell MD
- Subjects
- Cell Line, Tumor, Cell Survival, Dipeptidases antagonists & inhibitors, Dipeptidases metabolism, Dipeptidyl-Peptidases and Tripeptidyl-Peptidases antagonists & inhibitors, ErbB Receptors metabolism, HEK293 Cells, HeLa Cells, Humans, Liver Cirrhosis complications, Liver Cirrhosis enzymology, Liver Neoplasms enzymology, Liver Neoplasms etiology, Phosphatidylinositol 3-Kinases metabolism, Proto-Oncogene Proteins c-akt metabolism, Signal Transduction, Apoptosis, Cell Proliferation, Dipeptidyl-Peptidases and Tripeptidyl-Peptidases metabolism, Epidermal Growth Factor metabolism
- Abstract
Dipeptidyl peptidase IV (DPP4), DPP8, DPP9, and fibroblast activation protein (FAP), the four proteases of the DPP4 gene family, have unique peptidase and extra-enzymatic activities that have been implicated in various diseases including cancers. We report here a novel role of DPP9 in regulating cell survival and proliferation through modulating molecular signaling cascades. Akt (protein kinase B) activation was significantly inhibited by human DPP9 overexpression in human hepatoma cells (HepG2 and Huh7) and human embryonic kidney cells (HEK293T), whereas extracellular signal-regulated kinases (ERK1/2) activity was unaffected, revealing a pathway-specific effect. Interestingly, the inhibitory effect of DPP9 on Akt pathway activation was growth factor dependent. DPP9 overexpression caused apoptosis and significantly less epidermal growth factor (EGF)-mediated Akt activation in HepG2 cells. However, such inhibitory effect was not observed in cells stimulated with other growth factors, including connective tissue growth factor, hepatic growth factor, insulin or platelet-derived growth factor-BB. The effect of DPP9 on Akt did not occur when DPP9 enzyme activity was ablated by either mutagenesis or inhibition. The phosphatidylinositol 3-kinase (PI3K)/Akt pathway is a major downstream effector of Ras. We found that DPP9 and DPP8, but not DPP4 or FAP, associate with H-Ras, a key signal molecule of the EGF receptor signaling pathway. These findings suggest an important signaling role of DPP9 in the regulation of survival and proliferation pathways., (©2011 AACR.)
- Published
- 2011
- Full Text
- View/download PDF
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