Education

Biography

Educated at Imperial College London, UK , Post-doctoral experience in Benoit de Crombrugghe lab , Houston TX and Ramirez Lab in NY. Appointed, Research fellow with Terry Partridge at the MRC Clinical Sciences Centre, Hammersmith, UK.

First academic appointment as Senior Lecturer with Patrick. Maxwell at Renal Medicine, Hammersmith Hospital and then Reader in Matrix Biology. Moved to Kennedy Institute at Imperial College London and then University of Oxford as part of Nuffield department of Orthopaedics Rheumatology  and Musculoskeletal Sciences.

Appointed to Chair of Matrix Biology at the University of Liverpool to head a Transgenic Unit and animal models of musculoskeletal diseases.

Research Interest

My main scientific interest focuses on understanding how the body maintains homeostasis of connective tissues and how this is impaired in disease. Three concurrent lines of investigations are the focus of my research group:

1. Transcriptional regulation of major matrix proteins such as collagen type I and Aggrecan. Identification of non-coding sequences that drive expression in a cell specific manner.
2. The use of tissue-specific promoters to deliver modulating proteins to prevent or repair connective tissue diseases.
3. Characterisation of cells involved in fibrosis using transgenes as a read out in animal models of fibrosis.

1- Transcriptional regulation of major matrix proteins such as collagen type I and the identification of non-coding sequences that drive expression in a cell specific manner

In the last ten years my major effort has been dedicated to unraveling the transcriptional regulation of the alpha 2 chain of collagen type I, as a paradigm for collagen transcription. The major contribution in this area was the discovery of an enhancer region, which is critical for the expression of the gene, located some 17kb upstream of the start of transcription. The transgenic lines that were made harbouring this enhancer linked to various reporter genes have provided an excellent surrogate marker for fibrosis and are now used by many pharmaceutical companies to test a range of anti-fibrotic drugs in vivo. I showed that this enhancer was evolutionarily conserved between rodent and human and required the cooperation with the proximal promoter element to drive complete spatial and temporal expression of the gene. This discovery has also led to the delineation of tissue specific elements such as skin fibroblasts, smooth muscle cells and others that direct collagen expression in specific cell lineages which we are currently pursuing.

2- The use of tissue-specific promoters to deliver modulating proteins to prevent or repair connective tissue diseases

Extracellular matrix (ECM) homeostasis is regulated by the catabolic activity of proteinases; chiefly amongst them are the metalloproteinases (MMPs) that have the combined capacity to degrade all components of the ECM.  MMPs are in turn regulated by their inhibitors (TIMPs). We have previously shown that in fibrotic conditions such as Systemic Sclerosis, fibroblasts from these patients express a significant reduction in MMPs and concomitant increase in TIMPs. In order to understand the role of the MMPs and their inhibitors in connective tissue homeostasis, we have generated transgenic mice over expressing specific MMPs or TIMPs either in fibroblasts, using fibroblast-specific collagen type I promoter, or chondrocyte-specific aggrecan promoter that we have identified.

In the former animal model, we investigate the effect of MMPs, such as MMP14 in repair, using injury models of fibrosis which are established in our laboratory such as skin wound healing, bleomycin in lung fibrosis and unilateral uretral ligation in kidney fibrosis.

Over expression of TIMPs in chondrocytes is used in collaboration with Professor Nagase, Kennedy Institute of Rheumatology, to test the protective role of the proteinase inhibitors in vivo. Mutants of TIMP which can discriminate between different classes of proteinases are used to identify the main enzymes which degrade articular cartilage in mouse models of osteoarthritis. These models are being used in collaboration with Prof. Andrew Pitsillides to decipher the molecular pathway between naturally occurring and injury induced arthritis.

3- Characterisation of cells involved in fibrosis using transgenes as a read out in animal models of fibrosis

1. The role of bone marrow cells in repair and tissue fibrosis:
   
   Regardless of the primary cause of the tissue fibrosis, the hallmark of scarring is the excessive deposition of collagen type I molecules. Our current working hypothesis suggests that tissue fibrosis is the inability to terminate repair. Under this hypothesis, we have investigated the role of bone marrow derived cells in the deposition of matrix proteins in liver, kidney and skeletal muscle fibrosis as well as following heart infarct and showed that these cells do contribute to scarring but the level of such contribution is dependent on the injury model.
2. The role of fetal mesenchymal cells in maternal tissue repair:
   
   In collaboration with Professor Nicholas Fisk, we have investigated fetal cells involvement in tissue repair during and following pregnancy. The use of our collagen type I transgenic model showed that these cells are not only recruited to the site of injury but they synthesise collagen type I indicating their involvement in the repair process in the maternal tissue. We are currently addressing the mechanism underlying such recruitment, the phenotype of such cells and how these cells can be utilized for repair.
   This programme of proposed work will significantly improve our understanding and will identify therapeutic targets and molecules which would be beneficial in connective tissue diseases. Ultimately such knowledge may allow therapeutic strategies aimed at increasing the repair process by altering decisions by these cells concerning the balance between scarring / fibrosis and restoration of normal architecture

Scientific Activities:

Employment

Publications

1. Poulet B, Liu K, Plumb D, Vo P, Shah M, Staines K, Sampson A, Nakamura H,Nagase H, Carriero A, Shefelbine S, Pitsillides AA, Bou-Gharios G. Overexpressionof TIMP-3 in Chondrocytes Produces Transient Reduction in Growth Plate Length butPermanently Reduces Adult Bone Quality and Quantity. PLoS One. 2016 Dec 21;11(12):e0167971.
2. Hirai M, Kitahara H, Kobayashi Y, Kato K, Bou-Gharios G, Nakamura H, Kawashiri S. Regulation of PD-L1 expression in a high-grade invasive human oral squamouscell carcinoma microenvironment. Int J Oncol. 2017 Jan;50(1):41-48.
3. Sacilotto N, Chouliaras KM, Nikitenko LL, Lu YW, Fritzsche M, Wallace MD, Nornes S, García-Moreno F, Payne S, Bridges E, Liu K, Biggs D, Ratnayaka I, Herbert SP, Molnár Z, Harris AL, Davies B, Bond GL, Bou-Gharios G, Schwarz JJ, De Val S. MEF2 transcription factors are key regulators of sprouting angiogenesis. Genes Dev. 2016 Oct 15;30(20):2297-2309.
4. Kitahara H, Hirai M, Kato K, Bou-Gharios G, Nakamura H, Kawashiri S. Eribulinsensitizes oral squamous cell carcinoma cells to cetuximab via induction ofmesenchymal-to-epithelial transition. Oncol Rep. 2016 Dec;36(6):3139-3144.
5. Ikeda T, Fragiadaki M, Shi-Wen X, Ponticos M, Khan K, Denton C, Garcia P,Bou-Gharios G, Yamakawa A, Morimoto C, Abraham D. Data on CUX1 isoforms inidiopathic pulmonary fibrosis lung and systemic sclerosis skin tissue sections.Data Brief. 2016 Aug 10;8:1377-80.
6. Javaheri B, Hopkinson M, Poulet B, Pollard AS, Shefelbine SJ, Chang YM, Francis-West P, Bou-Gharios G, Pitsillides AA. Deficiency and Also Transgenic. Overexpression of Timp-3 Both Lead to Compromised Bone Mass and Architecture In Vivo. PLoS One. 2016 Aug 2;11(8):e0159657.
7. Becker PW.,Sacilotto‎ N.,  Nornes S., Neal A., Thomas, M., Liu K., Preece C., Ratnayaka I., Davies B., Bou-Gharios G., and Sarah De Val. An intronic Flk1 enhancer directs arterial-specific expression via RBPJ-mediated venous repression. ArteriosclerThrombVasc Biol. 2016 Jun;36(6):1209-19.
8. McDermott BT, Ellis S, Bou-Gharios G, Clegg PD, Tew SR. RNA binding proteins regulate anabolic and catabolic gene expression in chondrocytes. Osteoarthritis Cartilage. 2016 Feb 4.pii: S1063-4584(16)01007-4.
9. Moschidou D, Corcelli M, Hau KL, Ekwalla VJ, Behmoaras JV, De Coppi P, David AL, Bou-Gharios G, Cook HT, Pusey C, Fisk NM, Guillot PV. Human Chorionic Stem Cells: Podocyte Differentiation and Potential for the Treatment of Alport Syndrome. Stem Cells Dev. 2016 1;25(5):395-404.
10. Gardiner MD, Vincent TL, Driscoll C, Burleigh A, Bou-Gharios G, Saklatvala J, Nagase H, Chanalaris A. Transcriptional analysis of micro-dissected articular cartilage in post-traumatic murine osteoarthritis. Osteoarthritis Cartilage. 2015 Apr;23(4):616-28.
11. Ponticos M, Papaioannou I, Xu S, Holmes AM, Khan K, Denton CP, Bou-Gharios G, Abraham DJ. Failed degradation of JunB contributes to overproduction of type Icollagen and development of dermal fibrosis in patients with systemic sclerosis.Arthritis Rheumatol. 2015 Jan;67(1):243-53.
12. Sakai K, Jawaid S, Sasaki T, Bou-Gharios G, Sakai T. Transforming growthfactor-β-independent role of connective tissue growth factor in the development of liver fibrosis. Am J Pathol. 2014 Oct;184(10):2611-7
13. Lim NH, Meinjohanns E, Bou-Gharios G, Gompels LL, Nuti E, Rossello A, Devel L, Dive V, Meldal M, Nagase H.  In vivo imaging of matrix metalloproteinase 12 and matrix metalloproteinase 13 activities in the mouse model of collagen-induced arthritis.. Arthritis Rheumatol. 2014 Mar;66(3):589-98.
14. Lim NH, Meinjohanns E, Meldal M, Bou-Gharios G, Nagase H.In vivo imaging of MMP-13 activity in the murine destabilised medial meniscus surgical model of osteoarthritis. Osteoarthritis Cartilage. 2014 Jun;22(6):862-8.
15. Sakai K, Jawaid S, Sasaki T, Bou-Gharios G, Sakai T.Transforming growth factor-β-independent role of connective tissue growth factor in the development of liver fibrosis. Am J Pathol. 2014 Oct;184(10):2611-7.
16. Ponticos M, Papaioannou I, Xu S, Holmes AM, Khan K, Denton CP, Bou-Gharios G, Abraham DJ Failed degradation of JunB contributes to overproduction of type I collagen and development of dermal fibrosis in patients with systemic sclerosis Arthritis Rheumatol. 2015 Jan;67(1):243-53.
17. Gardiner MD, Vincent TL, Driscoll C, Burleigh A, Bou-Gharios G, Saklatvala J, Nagase H, Chanalaris A. Transcriptional analysis of micro-dissected articular cartilage in post-traumatic murine osteoarthritis.Osteoarthritis Cartilage. 2015 Apr;23(4):616-28.
18. Lo Cascio L., Liu K., Nakamura H., Chu G., Lim N  G., Chanalaris A., Saklatvala J., Nagase H.,andBou-Gharios,G.Generation of cartilage specific luciferase reporter mouse and inducible Crerecombinase activity. Genesis. 2014 Feb;52(2):110-9.
19. Crowe J, Aubareda A, McNamee K, Przybycien PM, Lu X, Williams RO, Bou-Gharios G, Saklatvala J, Dean JL. Heat shock protein b1-deficient mice display impaired wound healing PLoS One. 2013 Oct 15;8(10)
20. LindahlGE, Stock CJ, Shi-Wen X, Leoni P, Sestini P, Howat SL, Bou-Gharios G, Nicholson AG, Denton CP, Grutters JC, Maher TM, Wells AU, Abraham DJ, Renzoni EA.Microarray profiling reveals suppressed interferon stimulated gene program in fibroblasts from scleroderma-associated interstitial lung disease. Respir Res. 2013 Aug 2;14:80.
21. SacilottoN ,Monteiro M.,  Fritzsche M., Becker P., Sanchez-Del-Campo L., Liu, K., Pinheiro P., Ratnayaka I., Davies B., Goding C., Patient  R., Bou-GhariosG.,and De Val  S. Analysis of Dll4 regulation reveals a novel combinatorial role for Sox and Notch in arterial developmentPNAS Jul 16;110(29):11893-8
22. Seppanen E, Roy E, Ellis R, Bou-Gharios G, Fisk NM, Khosrotehrani K.Distant mesenchymal progenitors contribute to skin wound healing and produce collagen: evidence from a murine fetal microchimerism model.PLoS One. 2013 May 1;8(5).
23. Rodero MP, Legrand JM, Bou-Gharios G, Khosrotehrani K. Wound-associated macrophages control collagen 1α2 transcription during the early stages of skin wound healing. ExpDermatol. 2013 Feb;22(2):143-5.
24. Jones GN, Moschidou D, Puga-Iglesias TI, Kuleszewicz K, Vanleene M, Shefelbine SJ, Bou-Gharios G, Fisk NM, David AL, De Coppi P, Guillot PV. Ontological differences in first compared to third trimester human fetal placental chorionic stem cells. PLoS One. 2012;7(9):e43395
25. Seppanen EJ, Hodgson SS, Khosrotehrani K, Bou-Gharios G, Fisk NM. Fetal Microchimeric Cells in a Fetus-Treats-Its-Mother Paradigm Do Not Contribute to Dystrophin Production in Serially Parous mdx Females. Stem Cells Dev. 2012 Oct 10;21(15):2809-16.
26. Fragiadaki M, Hill N, Hewitt R, Bou-Gharios G, Cook T, Tam FW, Domin J, Mason RM. Hyperglycemia Causes Renal Cell Damage via CCN2-Induced Activation of the TrkA Receptor: Implications for Diabetic Nephropathy. Diabetes. 2012 Sep;61(9):2280-8.
27. Finnson KW, Chi Y, Bou-Gharios G, Leask A, Philip A. TGF-ß signaling in cartilage homeostasis and osteoarthritis. Front Biosci (Schol Ed). 2012 Jan1;4:251-68.
28. Fragiadaki M, Witherden AS, Kaneko T, Sonnylal S, Pusey CD, Bou-Gharios G, and Mason RM.Interstitial fibrosis is associated with increased COL1A2 transcription in AA-injured renal tubular epithelial cells in vivo.  Matrix Biol. 2011Sep;30(7-8):396-403.
29. Fragiadaki M, Ikeda T, Witherden A, Mason RM, Abraham D, and Bou-Gharios G. High doses of TGF-β potently suppress type I collagen via the transcription factor CUX1. MolBiol Cell. 2011 1;22(11):1836-44.
30. Bou-Gharios G, Amin F, Hill P, Nakamura H, Maxwell P, Fisk NM.Microchimeric fetal cells are recruited to maternal kidney following injury and activate collagen type I transcription. Cells Tissues Organs. 2011;193(6):379-92.
31. Vanleene M, Saldanha Z, Cloyd KL, Jell G, Bou-Gharios G, Bassett JH, Williams GR, Fisk NM, Oyen ML, Stevens MM, Guillot PV, Shefelbine SJ. Transplantation of human fetal blood stem cells in the osteogenesisimperfecta mouse leads to improvement in multiscale tissue properties. Blood. 2011 Jan 20;117(3):1053-60.
32. Higashiyama R, Nakao S, Shibusawa Y, Ishikawa O, Moro T, Mikami K, Fukumitsu H, Ueda Y, Minakawa K, Tabata Y, Bou-Gharios G, Inagaki Y. Differential contribution of dermal resident and bone marrow-derived cells to collagen production during wound healing and fibrogenesis in mice. J Invest Dermatol. 2011 Feb;131(2):529-36.
33. Lee ES, Bou-Gharios G,Seppanen E, Khosrotehrani K, Fisk NM.Fetal stem cell microchimerism natural-born healers or killers?Mol Hum Reprod. 2010 Nov;16(11):869-78.
34. Sonnylal S, Shi-Wen X, Leoni P, Naff K, Van Pelt CS, Nakamura H, Leask A, Abraham D, Bou-Gharios G, de Crombrugghe B. Selective expression of connective tissue growth factor in fibroblasts in vivo promotes systemic tissue fibrosis. Arthritis Rheum. 2010 May;62(5):1523-32.
35. Higashiyama R, Moro T, Nakao S, Mikami K, Fukumitsu H, Ueda Y, Ikeda K, Adachi E, Bou-Gharios G, Okazaki I, Inagaki Y. Negligible contribution of bone marrow-derived cells to collagen production during hepatic fibrogenesis in mice.  Gastroenterology. 2009 Oct;137(4):1459-66.e1
36. Ponticos M, Holmes AM, Shi-wen X, Leoni P, Khan K, Rajkumar VS, Hoyles RK, Bou-Gharios G, Black CM, Denton CP, Abraham DJ, Leask A, Lindahl GE. Pivotal role of connective tissue growth factor in lung fibrosis: MAPK-dependent transcriptional activation of type I collagen. Arthritis Rheum. 2009 Jul;60(7):2142-55.
37. Ponticos M, Harvey C, Ikeda T, Abraham D, Bou-Gharios G.JunB mediates enhancer/promoter activity of COL1A2 following TGF-{beta} induction. Nucleic Acids Res. 2009 Sep;37(16):5378-89.
38. Guillot, PV., H. Cook, T., Pusey C., Fisk, NM.,  Harten, S., Moss J., Shore, I., and Bou-Gharios G. Transplantation of human fetalmesenchymal stem cells improves glomerulopathy in a collagen type Ia2 deficient mouse J Pathol. 2008 214(5):627-36.
39. vanAmerongen M, Bou-Gharios G, Popa E, van Ark J, Petersen A, van Dam G, van Luyn M, and  Harmsen M.  Bone marrow-derived myofibroblasts contribute functionally to scar formation after myocardial infarction J Pathol. 2008 214(3):377-86.
40. Guillot PV, Abass O, Bassett JH, Shefelbine SJ, Bou-Gharios G, Chan J, Kurata H, Williams GR, Polak J, Fisk NM. Intrauterine transplantation of human fetalmesenchymal stem cells from first trimester blood repairs bone and reduces fractures in osteogenesisimperfecta mice Blood. 2008 Feb 1;111(3):1717-25.
41. Shi-Wen X, Kennedy L, Renzoni EA, Bou-Gharios G, du Bois RM, Black CM, Denton CP, Abraham DJ, Leask A. Endothelin is a downstream mediator of profibrotic responses to transforming growth factor beta in human lung fibroblasts. Arthritis Rheum. 2007 56(12):4189-94.)
42. Shi-Wen X, Renzoni EA, Kennedy L, Howat S, Chen Y, Pearson JD, Bou-Gharios G,Dashwood MR, du Bois RM, Black CM, Denton CP, Abraham DJ, Leask A.  Endogenous endothelin-1 signaling contributes to type I collagen and CCN2 overexpression in fibrotic fibroblasts.  Matrix Biol. 2007 26(8):625-32.
43. Alexakis, C., Partridge, T. and Bou-Gharios, G. Implication of the satellite cell in dystrophic muscle fibrosis: a self perpetuating mechanism of collagen over-production. Am J Physiol- Cell Physiol. 2007 293(2):C661-9.
44. Rajkumar VS, Shiwen X, Bostrom M, Leoni P, Muddle J, Ivarsson M, Gerdin B, Denton CP, Bou-Gharios G, Black CM, Abraham DJ. Platelet-derived growth factor-beta receptor activation is essential for fibroblast and pericyte recruitment during cutaneous wound healing. Am J Pathol. 2006 169(6):2254-65.
45. Russo, F, Alison, M., Bigger, B, Amofah, Y., Florou, E., Amin, F, Bou-Gharios, G. Jeffery, R., Iredale, J and Forbes, S. The bone marrow functionally contributes to liver fibrosis. Gastroenterology. 2006 Jun;130(6):1807-21.
46. Menon RP, Menon MR, Shi-Wen X, Renzoni E, Bou-Gharios G., Black CM, Abraham DJ. Hammerhead ribozyme-mediated silencing of the mutant fibrillin-1 of tight skin mouse: Insight into the functional role of mutant fibrillin-1. Exp Cell Res. 2006 15;312(9):1463-74.)
47. Xi-Wen X, Stanton L, Kennedy L, Pala D, Chen Y, Howat SL, Renzoni EA, Carter DE, Bou-Gharios G., Stratton R, Pearson JD, Beier F, Lyons KM, Black CM, Abraham DJ, Leask A. CCN2 is necessary for adhesive responses to TGFbeta 1 in embryonic fibroblasts. J Biol Chem. 2006 21;281(16):10715-26.
48. Roufosse C, Bou-Gharios G.,Prodromidi E, Alexakis C, Jeffery R, Khan S, Otto WR, Alter J, Poulsom R, Cook HT. Bone Marrow-Derived Cells Do Not Contribute Significantly to Collagen I Synthesis in a Murine Model of Renal Fibrosis. J Am SocNephrol. 2006 17(3):775-82.
49. Ramirez, F. Tanaka, S., and Bou-Gharios, G.,Transcriptional regulation of the human alpha2(I) collagen gene (COL1A2), an informative model system to study fibrotic diseases. Matrix Biol. 2006 Aug;25(6):365-72.
50. Chen Y, Shi-Wen X, van Beek J, Kennedy L, McLeod M, Renzoni EA, Bou-Gharios G., Wilcox-Adelman S, Goetinck PF, Eastwood M, Black CM, Abraham DJ, Leask A.   Matrix Contraction by Dermal Fibroblasts Requires Transforming Growth Factor-{beta}/Activin-Linked Kinase 5, HeparanSulfate-Containing Proteoglycans, and MEK/ERK: Insights into Pathological Scarring in Chronic Fibrotic Disease.  Am J Pathol. 2006 167(6):1699-711
51. Antoniv, T., Tanaka, T., De Val, S., Liu, K., Wells, D., Ramirez F., Bou-Gharios G., Identification of a repressor in the first intron of the human alpha 2(I) collagen gene (COL1A2) J Biol Chem. 2005 21;280(42):35417-23.
52. Inagaki Y, Kushida M, Higashi K, Itoh J, Higashiyama R, Hong YY, Kawada N, Namikawa K, Kiyama H, Bou-Gharios G., Watanabe T, Okazaki I, Ikeda K.  Cell Type-Specific Intervention of Transforming Growth Factor beta/SmadSignaling Suppresses Collagen Gene Expression and Hepatic Fibrosis in Mice. Gastroenterology. 2005 129(1):259-68.
53. Morrison J, Palmer DB, Cobbold S, Partridge T, Bou-Gharios G., Effects of T-lymphocyte depletion on muscle fibrosis in the mdx mouse. Am J Pathol. 2005 166(6):1701-10.
54. Alexakis C, Maxwell P, Bou-Gharios G. Organ-Specific Collagen Expression: Implications for Renal Disease.  Nephron ExpNephrol. 2005 Nov 10;102(3-4):e71-e75
55. Denton CP, Lindahl GE, Khan K, Shiwen X, Ong VH, Gaspar NJ, Lazaridis K, Edwards DR, Leask A, Eastwood M, Leoni P, Renzoni EA, BouGharios G., Abraham DJ, Black CM.  Activation of key profibrotic mechanisms in transgenic fibroblasts expressing kinase-deficient type II Transforming growth factor-beta receptor (T-beta-RII{delta}k). J Biol Chem. 2005 22;280(16):16053-65.
56. Morrison, J., Partridge, TA., Bou-Gharios, G. Nude mutation influences limb development. Matrix Biol. 2005 23: 535-42.
57. Lu QL, Rabinowitz A, Chen YC, Yokota T, Yin H, Alter J, Jadoon A, Bou-Gharios G., Partridge T. Systemic delivery of antisense oligoribonucleotide restores dystrophin expression in body-wide skeletal muscles. ProcNatlAcadSci U S A. 2005 4;102(1):198-203.

Book Chapter:

1. “Characterisation of mesenchymal-fibroblast cells using the Col1a2 promoter/enhancer" Ian M H Li, Amy L Horwell, Grace Chu, Benoit de Crombrugghe and George Bou-Gharios
2. Laure Rittié, Ph.D. Editor, Fibrosis: Methods and protocols, Methods in Molecular Biology Series, Springer, New-York.
3. Bou-Gharios, G. and de Crombrugghe, B. Type I Collagen: Structure, Synthesis and Regulation  in PRINCIPLES OF BONE BIOLOGY, 3rd Edition, 2008. Edited by Drs. John Bilezikian, Larry Raisz, and Jack Martin.