Ann Harris Group
Regulation of expression of the CFTR Gene. The cystic fibrosis transmembrane conductrance regulator (CFTR) gene shows a complex pattern of expression, with temporal and spatial regulation that is not accounted for by elements in the promoter. The gene is expressed at high levels in intestinal epithelia and at much lower levels in cells within the respiratory epithelium, where the tissue-specific control mechanisms are apparently different. Using a combined approach of DNase1 hypersensitive site (DHS) mapping and comparative genomics we identified candidate regulatory elements flanking the gene and within introns. The major focus of current research is to determine 1) the tissue-specific trans-acting factors that interact with these cis-acting elements; 2) how these elements interact with the CFTR gene promoter and 3) the alterations in chromatin structure and modifications that are associated with CFTR expression.
Function of MUC4 airway mucin.
The normal health of all epithelial surfaces in the human body is dependent on the presence of a surface layer of adequately hydrated mucus. The major protein components of this mucus layer are mucus glycoproteins, mucins, which may either be attached to the cell membrane or secreted from the epithelial surface. In addition to serving as a barrier that protects and lubricates the epithelium, mucins function as adhesion/anti-adhesion molecules, in invasion and metastasis and in intracellular signaling. We are working on the major membrane-associated mucin in the airway, MUC4, to determine its role in airway epithelial function in health and disease. We predict that this molecule acts as an extracellular sensor of airway environment and can have a profound effect on intracellular processes.
Function of MUC6 pancreatic mucin.
The normal functions of mucin glycoproteins may be usurped by cancer cells. We are dissecting the biological properties of an abundant secreted mucus glycoprotein, MUC6, that is found at several organ sites, including the pancreas, but about which very little is currently known. We are evaluating the hypothesis that MUC6 functions in the normal pancreatic duct to maintain epithelial integrity, protect the epithelium from environmental insults and contribute to the repair of epithelial damage. However, in the early stages of pancreatic cancer, aberrant expression of MUC6 may contribute to progression of this devastating disease.
Collagen XV as a tumor suppressor.
Collagen XV is a proteoglycan found in the basement membrane zones of a number of tissues but its precise functions remain to be elucidated. Our published data on cervical cancer epithelial cells support the hypothesis that human collagen XV is a dose-dependent suppressor of tumorigenicity. We predict that the hypothesis will be applicable to other epithelial tumors and are currently testing our hypothesis in the epithelium of the pancreatic duct. Pancreatic adenocarcinomas are generally highly metastatic and are associated with an extremely poor prognosis in comparison to many other cancers. The initiating events that underlie the escape of a pancreatic tumor cell, from its site of origin in the ductal epithelium, through the basement membrane of the pancreatic duct are fundamental to the progression of the pancreatic adenocarcinoma. Elucidation of the key molecular events involved in this escape are a central focus of this project.
Selected Recent Publications
Lewandowska M.A., Costa F., Bischof J.M., Williams S.H., Soares M.B., and Harris, A. (2010) Multiple mechanisms influence regulation of the cystic fibrosis transmembrane conductance regulator gene promoter. Am J. Respir Cell Mol Bio. 43(3): 334-41.
Ott C.J., Blackledge N.P., Leir S.H., Harris A. (2009) Novel regulatory mechanisms for the CFTR gene. Biochem. Soc. Trans. 37:843-8.
Blackledge N.P., Ott C.J., Gillen A.E., Harris A. (2009) An insulator element 3' to the CFTR gene binds CTCF and reveals an active chromatin hub in primary cells. Nucleic Acids Res. March;37(4): 1086-94.
Ott C., Suszko M., Blackledge N.P., Wright J.E., Crawford G.E., Harris A. (2009) A complex intronic enhancer regulates expression of the CFTR gene by direct interaction with the promoter. J. Cell and Mol. Med. Apr;13(4):680-92.
Kotzamanis G., Abdulrazzak H., Gifford-Garner J., Haussecker P.L., Cheung W., Harris A., Kotsinas A., Gorgoulis V., Huxley C. (2008) CFTR expression from a BAC carrying the complete human gene and associated regulatory elements. J. Cell and Mol. Med. (in press)
Evans J.R., Kelly D.L., Morris K.J., Arvide E.M., Harris A. (2008) RNA interference-mediated inhibition of hepatocyte nuclear factor 1alpha identifies target genes. Biochemica et Biophysica Acta May; 1779(5); 341-6.
Harris A., Harris H., Hollingsworth M.A. (2007) Complete suppression of tumor formation by high levels of basement membrane collagen. Mol Cancer Research 5(12): 1241-5.
Blackledge N.P., Carter E.J., Evans J.R., Lawson V., Rowntree R.K., Harris A. (2007) CTCF mediates insulator function at the CFTR locus. Biochem J. Dec 1; 408(2): 267-75.
Parry S., Hanisch F.G., Leir S.H., Sutton-Smith M., Morris H.R., Dell A., Harris A. (2006) N-Glycosylation of the MUC1 mucin in epithelial cells and secretions. Glycobiology Jul;16(7): 623-34.
Shiraga T., Winpenny J., Carter E., McCarthy V., Hollingsworth M.A., Harris A. (2005) Evaluation of potential regulatory elements in two DNase I hypersensitive sites in the MUC1 gene promoter. Exp. Cell Research. 308:41-52.
Palmai-Pallag T., Khodabukus N., Kinarsky L., Leir S-H., Sherman S., Hollingsworth M.A. and Harris A. (2005) The role of the SEA (sea urchin sperm protein, enterokinase and agrin) - module in cleavage of membrane-tethered mucins. FEBS J. 272:2901-11.
Leir S.H., Parry S., Palmai-Pallag T., Evans J.R., Morris H.R., Dell A., Harris A. (2005) Mucin glycosylation and sulphation in airway epithelial cells is not influenced by CFTR expression. Am. J. Resp. Cell and Mol. Biol. 32(5):453-61.
Parry S., Sutton-Smith M., Harrison D., Heal P., Hollingsworth M.A., Morris H.R., Dell A., Harris A. (2005) In vivo glycosylation of MUC6 mucin tandem repeats. Biochem. Biophys. Acta: General Subjects. 1722(1):77-83.
Disset A., Michot C., Harris A., Buratti E., Claustres M., Tuffery-Giraud S. (2004) A T3 allele in the CFTR gene exacerbates exon 9 skipping in vas deferens and epididymal cell lines and is associated with Congenital Bilateral Absence of Vas Deferens (CBAVD). Human Mutation. 25:72-81
Mouchel N., Henstra S.A., McCarthy V.A., Williams S.H, Phylactides M., Harris A. (2004) HNF1alpha is involved in regulation of expression of the CFTR gene. Biochemical Journal. 378: 909-918.
Williams S.H., Sahota V., Palmai-Pallag T., Tebbutt S.J., Walker J., Harris A. (2003) Evaluation of gene targeting by homologous recombination in ovine somatic cells. Molec. Reprod. and Devel. 66:115-125.
Broackes-Carter F., Williams S.H, Wong, P.L. Harris A. (2003) Alternative splicing of the ovine CFTR gene. Mammalian Genome. 14, 778ˆ787.
Williams S.H., Mouchel N., Harris A. (2003) A comparative genomic analysis of the cow, pig and human CFTR genes identifies potential intronic regulatory elements. Genomics 81: 628-639.
Mouchel N., Broackes-Carter F., Harris A. (2003) Alternative 5' exons of the CFTR gene show developmental regulation. Hum. Mol.Genet.12: 759-769.
Morris H.R., Dell A., Harris A. (2003) The contribution of tandem repeat number to the O-glycosylation of mucins. Glycobiology. 13: 265-277.