Liver Cell Biology
Lab head: Dr Nicholas Shackel
Location: Centenary Institute of Cancer Medicine and Cell Biology
To understand the molecular mechanisms regulating liver fibrosis aiming to developed novel therapeutic agents as well as developing new diagnostic investigations.
The current focus of the research group is to further understand the role of the glycoprotein CD147 in mediating basic and fundamental aspects of tissue inflammatory responses and carcinogenesis. Our group has made the fundamental discovery that CD147 mediates a process of inflammatory cell aggregation, which was previously not recognised that mediates inflammatory responses and carcinogenesis. Taking this novel discovery from the bench through to high impact papers is the current focus of the group.
Lab members: Dr Nicholas Shackel Dr Fiona Warner A/Prof Susan McLennnan Dr Annette Maczurek (Post-doctoral scientist) Aimei Lee (PhD Student) William d'Avigdor (PhD Student) Dr Emilia Prakoso (PhD Student) 1 new post-doctoral scientist starting in Nov 2012 2 new PhD students starting in 2013
Funding: In excess of $500,000
Research approach equipment: We use a range of techniques includes genomics analysis, confocal fluorescence, flow cytometry, Immunohistochemistry, quantitative PCR and Western blot analysis. Our group runs the facility for Gene Array Analysis with thin the Centenary Institute using both the latest Affymetrix Platform and Nimblegen
1. Rahman W, Huang P, Belov L, Chrisp JS, Christopherson RI, Stapelberg PM, Warner FJ, et al. Cluster of Differentiation (CD) Antibody Array Analysis of Human Liver Disease. Liver International 2012.
2. McCaughan GW, Gorrell MD, Bishop GA, Abbott CA, Shackel NA, McGuinness PH, Levy MT, et al. Molecular pathogenesis of liver disease: an approach to hepatic inflammation, cirrhosis and liver transplant tolerance. Immunological reviews 2000;174:172-191.
3. Yin JL, Shackel NA, Zekry A, McGuinness PH, Richards C, Putten KV, McCaughan GW, et al. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for measurement of cytokine and growth factor mRNA expression with fluorogenic probes or SYBR Green I. Immunology and cell biology 2001;79:213-221.
4. McGaughan GW, Shackel NA, Gorrell MD. Discussion on differential gene expression between chronic hepatitis B and C hepatic lesion. Gastroenterology 2001;121:1263-1264.
5. Shackel NA, McGuinness PH, Abbott CA, Gorrell MD, McCaughan GW. Insights into the pathobiology of hepatitis C virus-associated cirrhosis: analysis of intrahepatic differential gene expression. The American journal of pathology 2002;160:641-654.
6. Shackel NA, Gorrell MD, McCaughan GW. Gene array analysis and the liver. Hepatology 2002;36:1313-1325.
7. Shackel NA, Rockey DC. Intrahepatic gene silencing by RNA interference. Gastroenterology 2004;126:356-358; discussion 358-359.
8. Shackel NA, Rockey DC. Stem cells and liver disease: promise laced with confusion and intrigue. Gastroenterology 2004;127:346-348.
9. Shackel N, Rockey D. In pursuit of the "Holy Grail"--stem cells, hepatic injury, fibrogenesis and repair. Hepatology 2005;41:16-18.
10. Shackel NA, Seth D, Haber PS, Gorrell MD, McCaughan GW. The hepatic transcriptome in human liver disease. Comparative hepatology 2006;5:6.
11. Shackel NA, McCaughan GW, Warner FJ. Hepatocellular carcinoma development requires hepatic stem cells with altered transforming growth factor and interleukin-6 signaling. Hepatology 2008;47:2134-2136.
12. McCaughan GW, Shackel NA, Bertolino P, Bowen DG. Molecular and cellular aspects of hepatitis C virus reinfection after liver transplantation: how the early phase impacts on outcomes. Transplantation 2009;87:1105-1111.
13. Rauch A, James I, Pfafferott K, Nolan D, Klenerman P, Cheng W, Mollison L, et al. Divergent adaptation of hepatitis C virus genotypes 1 and 3 to human leukocyte antigen-restricted immune pressure. Hepatology 2009;50:1017-1029.
14. Eckersley-Maslin MA, Warner FJ, Grzelak CA, McCaughan GW, Shackel NA. Bone marrow stem cells and the liver: are they relevant? Journal of gastroenterology and hepatology 2009;24:1608-1616.
15. Song S, Shackel NA, Wang XM, Ajami K, McCaughan GW, Gorrell MD. Discoidin domain receptor 1: isoform expression and potential functions in cirrhotic human liver. The American journal of pathology 2011;178:1134-1144.
16. McCaughan GW, Shackel NA, Bowen DG. Liver transplantation and hepatitis C: will understanding the interleukin-28B polymorphisms improve outcomes? Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society 2011;17:219-221.
17. Bao W, Min D, Twigg SM, Shackel NA, Warner FJ, Yue DK, McLennan SV. Monocyte CD147 is induced by advanced glycation end products and high glucose concentration: possible role in diabetic complications. American journal of physiology. Cell physiology 2010;299:C1212-1219.
18. Shackel NA, McGuinness PH, Abbott CA, Gorrell MD, McCaughan GW. Insights into the Pathobiology of Hepatitis C Virus Associated Cirrhosis: Analysis of Intrahepatic Differential Gene Expression. American Journal of Pathology 2002;160:641-654.
Discovering Novel Biomarkers In Hepatocellular Carcinoma (HCC)
Primary supervisor: Nicholas Shackel
Discovering Novel Biomarkers In Hepatocellular Carcinoma (HCC)
Supervisors: Dr. Nick Shackel and A/Prof. Sue McLennan
Liver Cell Biology Centenary Institute http://www.centenary.org.au/p/ourresearch/liver/liverimmunobiology/LiverCellBiology/
Dr. Nick Shackel firstname.lastname@example.org 9565-6286
A/Prof. Sue McLennan email@example.com 9515-5185
Background Hepatocellular carcinoma (HCC) is the 4th most common human malignancy and the 3rd leading cause of cancer death. HCC almost invariably develops in the cirrhotic liver. Two of the most pressing clinical questions in HCC diagnosis and management are: (1) How to diagnose early HCC in cirrhotic individuals? and (2) How to predict the HCC behavior?
Microparticles (MPs): MPs are membrane bound vesicular structures that are readily found within the circulation and act in a hormone-like way [1-5]. They are released by cells in response to activation, malignant transformation or stress and are stable within the circulation. MPs are derived from the plasma membrane (shed microvesicles), produced by the endosomal pathway (exosomes), or arise from membrane blebs of apoptotic cells. In addition to proteins and lipids MPs are known to contain RNA (in particular micro RNAs) and DNA[1, 2, 6]. The profiling of nucleic acids within MP is now a new and novel area of research [4, 7-9].
CD147: Our group identified the glycoprotein CD147 as being important in liver injury and liver cancer development using gene array analysis. We pursued studies of this protein given its known roles in extra-hepatic malignancy, extra cellular matrix degradation and energy metabolism via lactate transportation[10-14]. Our studies have shown that CD147 is found in MPs and could be used as a prognostic marker in serum of HCC patients. Further, we have shown that MPs increase with disease severity and that CD147 positive MPs can activate hepatic stellate cells (HSC) in the liver, leading to increased fibrosis. Our in-vivo experiments have shown diminished HCC burden in a metastatic mouse model of HCC with anti-CD147 antibody intervention.
These observations combined with our track record in CD147 research, means we are well placed to characterise the relationship between CD147 positive MPs and the development and progression of liver disease from cirrhosis to HCC[10, 16].This honours proposal will take advantage of our access to patients at different stages of liver disease from which we will obtain plasma and serum samples to identify novel diagnostic and prognostic markers in MPs. We are uniquely placed to characterize novel gene expression in HCC, possess a demonstrated Track Record and have established methods to characterize novel gene targets.
1. MPs containing novel, uncharactized HCC specific nucleic acid sequences are present in the circulation of HCC affected individuals. We have labelled this HCC Nucleic Acid Signature and Signature could serve as a new class of previously unrecognised biomarkers for HCC.
2. Circulating MPs are increased in liver injury and are associated with progression and/or prognosis of liver injury through to cirrhosis and then HCC
3. Biomarkers, especially those within microparticles, will enable us to better understand HCC pathogenesis and prognosis, resulting in better treatment outcome for affected individuals
1. To identify novel biomarkers in HCC using Next Generation Sequencing (NGS).
2. To characterise MPs and identify novel biomarkers in MPs in human HCC.
3. To validate novel biomarkers on a large clinical cohort being screened for HCC.
4. To use an animal model of HCC to determine the functional role of MPs and CD147 in progressive fibrosis and HCC development.
This is a major funded project within our group and it is envisaged that the student undertaking this project would work as a part of a team including a PhD student and post-doctoral scientist. The student would adopt one of the aims outlined above as their primary area of research to complete as an honours whilst having the support of others working on the same area on the other aims of this project.
Skills: This project will utilise confocal microscopy, real-time RT-PCR, Next Generation Sequencing, mammalian cell culture, transfection, animal models, primary cell isolation and flow cytometry as well as immunohistochemistry techniques. It is envisaged that the student who undertakes this project will become proficient in all of these methods whilst being exposed to a number of other general laboratory techniques.
Contacts: For further information and to arrange a time to discuss the project details please contact Dr N. Shackel on 0434603129 or email firstname.lastname@example.org.
1. Amin, C., N. Mackman, and N.S. Key, Microparticles and cancer. Pathophysiology of haemostasis and thrombosis, 2008. 36(3-4): p. 177-83.
2. Rak, J., Microparticles in cancer. Seminars in thrombosis and hemostasis, 2010. 36(8): p. 888-906.
3. Martinez, M.C., et al., Microparticles: targets and tools in cardiovascular disease. Trends in pharmacological sciences, 2011. 32(11): p. 659-65.
4. Morel, O., et al., Microparticles: a critical component in the nexus between inflammation, immunity, and thrombosis. Seminars in immunopathology, 2011. 33(5): p. 469-86.
5. Liu, M.L. and K.J. Williams, Microvesicles: potential markers and mediators of endothelial dysfunction. Current opinion in endocrinology, diabetes, and obesity, 2012. 19(2): p. 121-7.
6. Turchinovich, A., et al., Characterization of extracellular circulating microRNA. Nucleic acids research, 2011. 39(16): p. 7223-33.
7. Jaiswal, R., et al., Microparticle-associated nucleic acids mediate trait dominance in cancer. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2012. 26(1): p. 420-9.
8. Qu, K.Z., et al., Circulating MicroRNAs as Biomarkers for Hepatocellular Carcinoma. J Clin Gastroenterol, 2011.
9. Jaiswal, R., et al., Microparticle conferred microRNA profiles - implications in the transfer and dominance of cancer traits. Molecular cancer, 2012. 11(1): p. 37.
10. Shackel, N.A., et al., Insights into the Pathobiology of Hepatitis C Virus Associated Cirrhosis: Analysis of Intrahepatic Differential Gene Expression. American Journal of Pathology, 2002. 160(2): p. 641-654.
11. Muramatsu, T. and T. Miyauchi, Basigin (CD147): a multifunctional transmembrane protein involved in reproduction, neural function, inflammation and tumor invasion. Histology and histopathology, 2003. 18(3): p. 981-7.
12. Iacono, K.T., et al., CD147 immunoglobulin superfamily receptor function and role in pathology. Experimental and molecular pathology, 2007. 83(3): p. 283-95.
13. Huet, E., et al., Role of emmprin/CD147 in tissue remodeling. Connective tissue research, 2008. 49(3): p. 175-9.
14. Weidle, U.H., et al., Cancer-related issues of CD147. Cancer genomics & proteomics, 2010. 7(3): p. 157-69.
15. Baran, J., et al., Circulating tumour-derived microvesicles in plasma of gastric cancer patients. Cancer immunology, immunotherapy : CII, 2010. 59(6): p. 841-50.
16. Bao, W., et al., Monocyte CD147 is induced by advanced glycation end products and high glucose concentration: possible role in diabetic complications. American journal of physiology. Cell physiology, 2010. 299(5): p. C1212-9.
Discipline: Infectious diseases and Immunology
Co-supervisors: Susan McLennan
Keywords: Cancer, Liver Cancer, Genomics
Contact: Email Nicholas Shackel