Eppley Institute for Research in Cancer and Allied Diseases

Michael H. Hollingsworth, Ph.D.
Professor

Back to Top

ACADEMIC DEGREES:
B.A., 1978, Biology, Wake Forest University, Winston-Salem, N.C.
Ph.D., 1982, Microbiology and Immunology, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, N.C.

Back to Top

Myself, at 559-8343, email: mahollin@unmc.edu or my secretary:
Contact Name: Julie Maloney, Admin Tech
Phone Numbers 402-559-8553

Back to Top

• GRANT SUPPORT (last 3 years):

Molecular Studies on the MUC1 Promoter
Principal Investigator: Michael A. Hollingsworth PhD
Agency: National Cancer Institute, National Institutes of Health; Type: RO1 CA 79580;
Period: 1998-2003
Summary: Characterize the MUC1 promoter for elements that control tissue specific expression

Mutagenesis Studies on the Post-Translational Processing of MUC1
Principal Investigator: Michael A. Hollingsworth PhD
Agency: National Cancer Institute, National Institutes of Health; Type: RO1 CA 57362;
Period: 1996-2005
Summary: Investigate structure and function of MUC1 by performing mutagenesis on the
MUC1 cDNA.

Molecular Analysis of Mucin-Type Glycosylation
Principal Investigator: Michael A. Hollingsworth PhD
Agency: National Cancer Institute, National Institutes of Health; Type: R01 CA 69234;
Period: 1996-2002
Summary: Characterize the tissue distribution and substrate specificity of several newly
cloned polypeptide GalNAc transferases.

Immunotherapy and Immunodiagnosis of Pancreatic Cancer
Principal Investigator: Michael A. Hollingsworth PhD
Agency:  National Cancer Institute, National Institutes of Health;
Type: SPORE in Gastrointestinal Cancer  1P50CA72712; Period:1997‑2004
Summary:  Develop and test new reagents for diagnosis and therapy of pancreatic cancer.

Back to Top

• PUBLICATIONS (last 3-5 years):

Tempero, R.M., Rowse, G.J., Gendler, S.J., and Hollingsworth, M.A. Anti-MUC1 antibodies cause neither autoimmunity nor tumor immunity in MUC1 transgenic mice. Int. J. Cancer 80: 595-599, 1999.

Morikane, K., Muto, T., Tempero, R., Sivinski, C., Nomoto, M., VanLith, M., Hollingsworth, M.A. Organ-specific pancreatic tumor growth properties and tumor immunity. Cancer Immunol. Immunother. 47:287-296, 1999.

Schwientek, T., Nomoto, M., Levery, S. B. Merkx, G., van Kessel, A. G. Bennett, E. P. Hollingsworth, M. A., Clausen, H. Control of O-glycan branch formation. Molecular cloning of human cDNA encoding a novel beta 1,6-N-acetylglucosaminyltransferase forming core 2 and core 4. J. Biol. Chem. 27:4504-4512, 1999.

Mack, D.R., Michail, S., Wei, S., McDougall, L., and Hollingsworth, M.A. Probiotics inhibit enteropathogenic Escheria coli adherence in vitro by inducing intestinal mucin gene expression. Am. J. Physiol. 276: G941-G950, 1999.

Reid, C.J., Burdick, M.D., Hollingsworth M.A., Harris A. CFTR expression does not influenceglycosylation of an epitope-tagged MUC1 mucin in colon carcinoma cell lines. Glycobiology 9: 389-398, 1999.

Akisawa, N., Nishimori, I., Iwamura, T., Onishi, S., and Hollingsworth, M.A High levels of ezrin expressed by human pancreatic adenocarcinoma cell lines with high metastatic potential. Biochem. Biophysic. Res. Comm. 258: 395-400, 1999.

Fujikawa-Adachi, K Nishimori, N, Sakamoto, S, Morita, M, Onishi, S, Yonezawa, S, Hollingsworth, MA. Identification of carbonic anhydrase IV and VI mRNA expression in human pancreas and salivary glands. Pancreas, 18:329-335,1999.

Nishimori, I., Kamakura, M., Fujikawa-Adachi, K., Nojima, M., Onishi, S., Hollingsworth, M. A., Harris, A. Cholecystokinin A and B receptor mRNA expression in human pancreas. Pancreas 19:109-113, 1999.

Bennett EP, Hassan H, Mandel U, Hollingsworth MA, Akisawa N, Ikematsu Y, Merkx G, van Kessel AG, Olufsson S, Clausen H. Cloning and Characterization of a Close Homologue of Human UDP-N-acetyl-a D-galactosamine: Polypeptide N-Acetylgalactosaminyltransferase T-3, designated GalNAc-T6: Evidence for Genetic but not Functional Redundancy. J Biol Chem 274:25362-25370, 1999.

Beum, P., Singh, P., Burdick, M., Hollingsworth, M.A., Cheng, P-W. Expression of core 2 beta 1, 6 N Acetylglucosaminyl transferase in a human pancreatic cancer cell line results in altered expression of MUC1 tumor associated epitopes. J. Biol. Chem. 274:24641-24648, 1999.

Hyde K., Harrison DA, Hollingsworth MA, Harris, A. Chloride-bicarbonate exchangers in the human fetal pancreas. Biochem. Biophys. Res. Comm. 263:315-321, 1999.

Satoh, S., Hinoda, Y., Hayashi, T., Burdick, M.D., Imai, K., Hollingsworth, M.A. Enhancement of metastatic properties of pancreatic cancer cells by MUC1 gene encoding an anti-adhesion molecule Int. J. Cancer 88:507-518, 2000.

Kirnarsky, L., Prakash, O., Vogen, S., Nomoto, M., Hollingsworth, M.A., and Sherman, S. Structural Effects of O-Glycosylation on a 15-residue Peptide from the Mucin Core Protein. Biochemistry, 39: 12076-12082, 2000.

Choudhury, A., Moniaux, N., Winpenny, J.P., Hollingsworth, M.A., Aubert, J.-P., and Batra, S. Human MUC4 Mucin cDNA and its variants in pancreatic cancer. J. Biochem. 128: 233-243, 2000.

Hassan, H., Reis, C.A., Bennett, E.P., Mirgorodskaya, E., Roepstorff, P., Hollingsworth, M.A., Burchell, J., Taylor-Papadimitriou, J., and Clausen, H. The lectin domain of UDP-N-acetyl-D-Galactosamine:Polypeptide-N-acetylgalactosaminyltransferase-T4 directs its glycopeptide specificities. J. Biol. Chem. 275:38197-38205, 2000

Kohsaki, T., Nishimori, N., Nakayama, H., Miyazaki, E., Enzan, H., Nomoto, M., Hollingsworth, M.A., Onishi, S. Expression of UDP-GalNAc:polypeptide N-acetylgalactosyltransferase isozymes T1 and T2 in human colorectal cancer. J. Gastroenterol. 35:840-848, 2000.

Morikane, K., Tempero, R., Sivinski, C., Gendler, S.J., and Hollingsworth, M.A. Influence of organ and cell type on MUC1-specific tumor immunity. International Immunology, 13:233-240, 2001.

Ryu, B., Jones, J., Hollingsworth, M. A., Hruban, R. H., Kern, S.E. Invasion-specific genes in malignancy: serial analysis of gene expression comparisons of primary and passaged cancers.  Cancer Res. 61:1833-1838, 2001.

Parry, S., Silverman, H., McDermott, K., Willis, A., Hollingsworth, M.A., and Harris, A.  Identification of the MUC1 proteolytic cleavage site in vivo.  Biochem. Biophysic. Res. Comm. 283: 715-720, 2001.

Kern, S., Hruban, R., Hollingsworth, M.A., Brand, R., Adrian, T. E., Jaffee, E., Tempero, M.A.  A white paper: The product of the pancreas cancer think tank.  Cancer Res. 61: 4923-4932, 2001.

Murphy, L., Abdel-Wahab, Y., Wang, Q., Knezetic, J., Permert, J., Hollingsworth, M., Adrian, T.  Receptors and ligands for autocrine growth pathways are up-regulated when pancreatic cancer cells are adapted to serum-free culture.  Pancreas 22: 293-298, 2001.

Silverman HS, Parry S, Sutton-Smith, M., Burdick MD, McDermott K, Reid, C.,  Batra SK, Morris, H.,  Hollingsworth MA, Dell, A., Harris A. in vivo Glycosylation of mucin tandem repeats. Glycobiology, 11:459-471, 2001.

McDermott, K.M., Crocker, P.R., Harris, A., Burdick, M.D., Hinoda, Y., Hayashi, T.,  Imai, K., and Hollingsworth, M.A.  Overexpression of MUC1 reconfigures the binding properties of tumor cells.  Int, J. Cancer 94:783-791, 2001.

Kitamura, N., Iwamura, T., Taniguchi, S., Yamanari, H., Kawano, K., Hollingsworth, M.A., Setoguchi, T.  High collagenolytic activity in spontaneously highly metatstatic variants derived from a human pancreatic cancer cell line (SUIT-2) in nude mice.  Clin Exp. Metastasis 18:561-571, 2001.

Andrianifahanana, M., Moniaux, N., Ringel, J., A., Schmied, B.M., Friess H., Hollingsworth, M.A., Aubert, J-P., and Batra, S.K. Mucin (MUC) gene expression in human pancreatic adenocarcinoma and chronic pancreatitis: A potential role of MUC4 as a tumor marker of diagnostic significance. Clin Can Res, 7:4033-4040, 2001

Khorrami A.M., Choudhury A., Andrianifahanana M., Varshney G.C., Bhattacharyya S.N., Hollingsworth M.A., Kaufman B, and Batra S.K. Purification and characterization of a human pancreatic adenocarcinoma mucin. J. Biochemistry, 131: 21-29, 2002.

Iacobuzio-Donahue, C., Maitra, A., Shen-Ong, G., van Heek, T., Ashfaq, R., Meyer, R., Walter, K., Berg, K., Hollingsworth, M., Cameron, J., Yeo, C., Kern, S., Goggins, M., Hruban, R.  Discovery of novel tumor markers of pancreatic cancer using global gene expression technology.  Am. J. Path. 160:1239-1249, 2002.

Swartz, M. Batra, S., Varshney, G., Hollingsworth, M., Yeo, C., Cameron, J., Wilentz, R., and Hruban, R.  MUC4 expression increases progressively in pancreatic intraepithelial neoplasia.  Am. J. Clin. Path. 117:791-796, 2002.

Sivinski, C.L., Kohlgraf, K.G., VanLith, M.L., Morikane, K., Tempero, R.M., and Hollingsworth, M.A.  Molecular requirements for CD8 mediated rejection of a MUC1-expressing pancreatic carcinoma:  Implications for tumor vaccines.  Cancer Immunol. Immunother. 51: 327-340, 2002.

VanLith, M.L., Kohlgraf, K.G., Sivinski, C.L., Tempero, R., and Hollingsworth, M.A.  MUC1-Specific Anti-Tumor Responses: Molecular Requirements for CD4 mediated responses. Int. Immunol. 14: 873-882, 2002.

Ryu, B., Jones, J., Blades, N. J., Parmigiani, G., Hollingsworth, M. A., Hruban, R. H., and Kern, S.E.  Relationships and differentially expressed genes among pancreatic cancers examined by large-scale serial analysis of gene expression. Cancer Res 62, 819-826, 2002. 

Schwientek, T., Bennett, E. P., Flores, C., Thacker, J., Hollmann, M., Reis, C. A., Behrens, J.,Mandel, U., Keck, B., Schafer, M. A., Haselman, K., Zubarev, R., Roepstorff, P., Burchell, J.M., Taylor-Papadimitriou, J. M., Hollingsworth, M.A., and Clausen, H. Functional conservation of subfamilies of putative UDP-N-acetylgalactosamine:polypeptide N-acetylgalactosaminyltransferases in Drosophila, Caenorhabditis elegans, and mammals.  One subfamily composed of l(2)35Aa is essential in Drosophila. J Biol Chem 277, 22623-22638, 2002.

Shiraga, T., Smith, D., Nuthall, H. N., Hollingsworth, M. A., and Harris, A. Identification of two novel elements involved in human MUC1 gene expression in vivo. Mol Med 8, 33-41, 2002.

Silverman, H. S., Sutton-Smith, M., McDermott, K., Heal, P., Leir, S. H., Morris, H. R., Hollingsworth, M. A., Dell, A., and Harris, A. The contribution of tandem repeat number to the O-glycosylation of mucins. Glycobiology 13, 265-277, 2003.

Mack, D. R., Ahrne, S., Hyde, L., Wei, S. & Hollingsworth, M. A. Extracellular MUC3 mucin secretion follows adherence of Lactobacillus strains to intestinal epithelial cells in vitro. Gut 52:827-33, 2003.

Kohlgraf, K.G., Gawron, A.J., Higashi, M., Meza, J.L., Burdick, M.D., Kitajima, S., Kelly, D.L., Caffrey, T.C., Hollingsworth, M.A  Contribution of the MUC1 tandem repeat and cytoplasmic tail to invasive and metastatic properties of a pancreatic cancer cell line. Cancer Research, 63:5011-5020, 2003.

Wen, Y., Caffrey, T. C., Wheelock, M. J., Johnson, K. R., Hollingsworth, M. A. Nuclear association of the cytoplasmic tail of MUC1 and b-catenin. J. Biol. Chem. 278: 38029-38039, 2003.

Kinarsky, L., Suryanarayanan G., Prakash O., Paulsen H., Clausen H., Hanisch F.G., Hollingsworth M.A., Sherman S.  Conformational studies on the MUC1 tandem repeat glycopeptides: implication for the enzymatic O-glycosylation of the mucin protein core. Glycobiology 13:929-939, 2003.

Iacobuzio-Donahue CA, Ashfaq R, Maitra A, Adsay NV, Shen-Ong GL, Berg K, Hollingsworth MA, Cameron JL, Yeo CJ, Kern SE, Goggins M, Hruban RH.  Highly expressed genes in pancreatic ductal adenocarcinomas: a comprehensive characterization and comparison of the transcription profiles obtained from three major technologies.  Cancer Res. 63:8614-22, 2003.

Hollingsworth, M.A., Swanson, B.J.  Mucins in Cancer:  Protection and Control of the Cell Surface.  Nature Reviews Cancer 4:45-60 2004.

Shibahara, H., Tamada, S., Higashi, M., Goto, M., Batra, S. K., Hollingsworth, M. A., Imai, K., and Yonezawa, S. MUC4 is a novel prognostic factor of intrahepatic holangiocarcinoma-mass forming type. Hepatology 39, 220-229, 2004. 

Choudhury, A., Moniaux, N., Ulrich, A.B., Schmied, B.M., Standoop, J., Pour, P.M., Gendler, S.J., Hollingsworth, MA., Aubert, J-P., Batra, S.K.  MUC4 mucin expression in human pancreatic tumours is affected by organ environment: the possible role of TGFb2.  Brit. J.Cancer 90: 657-664, 2004.

Back to Top

• PREVIOUS GRADUATE STUDENTS/POST-DOCTORAL FELLOWS (present location):

Graduate students trained in my laboratory (chairperson of supervisory committee)
1992-1998	Michael Burdick, PhD, Pathology and Microbiology
1994-1998	Richard Tempero, PhD, Pathology and Microbiology
1994-2001	Connie Sivinski, PhD candidate, Pathology and Microbiology
1996-2001	Kim McDermott, PhD candidate, Biochemistry and Molecular Biology
1996-2001	Michelle Van Lith, PhD candidate, Pathology and Microbiology
1997-2003	Karl Kohlgraf, PhD candidate, Pathology and Microbiology
1999-present	Yunfei Wen , PhD candidate, Biochemistry and Molecular Biology
1999-present	Xiao-Ling Shen, PhD candidate, Biochemistry and Molecular Biology
1999-present	Ganesh Suryanarayanan, PhD candidate, Biochemistry and Molecular Biology
2002-present	Benjamin Swanson, PhD candidate, Pathology and Microbiology
2002-present	Jarrod Treymayne, PhD candidate, Pathology and Microbiolgy
2002-present	Pankaj Singh, PhD candidate, Biochemistry and Molecular Biology
2002-present	Chunhui Yi, PhD candidate, Pathology and Microbiology

   

Postdoctoral fellows and visiting faculty trained
1988-1991	Surinder Batra, PhD, Postdoctoral Fellow
1989-1991	Wen-Ning Qi, MD, Postdoctoral Fellow
1991-1992	Judith Strawhecker, PhD, Postdoctoral Fellow
1992-1994	Isao Nishimori, MD-PhD, Visiting Assistant Professor (Kochi)
1993-1995	Marie Sutherlin, PhD, Postdoctoral Fellow
1993-1994	Troitza Tochova, PhD, Postdoctoral Fellow
1993-1996	Gurchuran Pahwa, PhD, Postdoctoral Fellow
1993-1995	Takeshi Iwamura, MD-PhD, Assistant Professor, (Miyazaki)
1996-1997	Yoshito Ikematsu, MD, Research Associate (Nagasaki)
1996	Yuji Hinoda MD, MD-PhD, Associate Professor (Sapporo)
1996-1998	Keita Morikane, MD, Research Associate (Tokyo)
1996-1999	John Winpenny, PhD, Welcome Advanced Research Fellowship (Newcastle)
1996-1998	Mitsuharu Nomoto, MD-PhD, Research Associate (Kagoshima)
1997-1999	Naoaki Akisawa, MD-PhD, Research Associate (Kochi)
1998-present	Shinichi Kitajima, MD-PhD, Research Associate (Kagoshima)
1998-2000	Shinichi Samishima, MD, Research Associate, (Tokyo)
1998-present	Beata Scholtz, PhD, Research Associate
1999	Helle Hassan, PhD, Research Associate
1999-present	David Smith, PhD, Research Associate
2000-present	Hiroaki Yasuda, MD, PhD, Research Associate
2001-present	Andrew Gawron, PhD
2001-present	Takuhiro Kohsaki, MD, PhD
2002-present	Hideaki Tsutsumida, MD, Research Associate

Back to Top

Summary of Current Research
The general subject of research in my laboratory is pancreatic cancer and other diseases of the pancreas (primarily pancreatitis and cystic fibrosis). We have used modern techniques of molecular biology, biochemistry, cell biology, and immunology to develop a comprehensive program of investigation into the biology of normal and diseased pancreatic ductal epithelialcells.

Several of the main projects in my laboratory center around the study of a complex mucin-like glycoprotein, MUC1, which is believed to play an important role in the normal function of pancreatic ductal epithelia and in the pathogenesis of pancreatic diseases Including pancreatic adenocarcinoma and cystic fibrosis. We are using the techniques of molecular and cellular biology to examine the regulation of expression and the mechanisms of post-translational processing of MUC1 in tumor cells and other disease conditions as compared to their normal cell counterparts. These studies also provide a paradigm to study basic aspects of the post-translational process (particularly O-glycosylation). We are developing and characterizing new monoclonal antibodies and tumor vaccine reagents for diagnostic and therapeutic uses that target known tumor associated antigens found on mucins. We are also studying the MUC1 promoter. Other projects in the lab involve the application of different cDNA cloning techniques to identify genes and their encoded proteins that are important to the process of development and differentiation of normal ductal cells in the human pancreas, and which may contribute to the pathogenesis of different diseases.

Specific funded projects.
Mutagenesis Studies on the Post-Translational Processing of MUC1 (NIH R01 CA 57362)
We cloned a cDNA for MUC1 from pancreatic adenocarcinoma cells in the late 1980's. MUC1 is an integral cell surface protein that has mucin-like characteristics and may play a role in cellular adhesion, anti-adhesion, and signal transduction events related to adhesion. The MUC1 protein receives a variety of distinct post-translational modifications (glycosylation, sulfation phosphorylation)in different normal cells and these are altered in tumors and other disease states. We are investigating the nature and biological functions of the different modifications in disease conditions. For example, altered glycoforms of MUC1 may play a role in the ability of tumor cells to metastasize.

Immunotherapy and Immunodiagnosis of Pancreatic Cancer (NIH SPORE 1P50CA72712)
MUC1 was originally identified as a tumor associated antigen of human adenocarcinomas. We are developing new reagents for detecting pancreatic cancer in the early stages (monoclonal antibodies) and investigating their usefulness by using a strain of mice transgenic for MUC1 (developed by collaborator Sandy Gendler). We are also designing and evaluating tumor vaccines. A key element of the project is to develop vaccines that break immunological tolerance, which is intact in the MUC1 transgenic mice. In arelated project (in collaboration with Sandy Gendler of Mayo Clinic Scottsdale, Olja Finn of Pittsburgh, and Don Kufe of Harvard),the MUC1 transgenic mice have been crossed with syngeneic mouse strains that harbor transforming genes under the control of tissue specific promoters. The resulting mice develop spontaneous tumors at specific organ sites (pancreas, prostrate, mammary gland) that express MUC1 in a manner that is similar to human tumors. This model system will be used to further test the usefulness of novel diagnostic and therapeutic strategies.

Molecular Studies on the MUC1 Promoter (NIH R01 CA 79580)
MUC1 expression is restricted to ductal epithelial cells and some populations of activated lymphocytes. It is overexpressed by tumors and is expressed by cell types in the pancreas that are the targets of a number of diseases (cancer, cystic fibrosis, pancreatitis). A relatively small fragment of DNA (10kb) conferred accurate expression temporally and spatially in a transgenic mouse strain. Hence, there is interest in defining the elements that direct cell specific expression of MUC1. We are using current in vivo and in vitro methods to investigate the role of elements and DNA structure in regulation of expression of this gene. Collaborators on this project include Ann Harris (Oxford), John Hilkens (Amsterdam), and Jim Maher (Mayo Clinic).

Molecular Analysis of Mucin-Type Glycosylation (NIH R01 CA 69234)
Together with Henrik Clausen (Copenhagen), we are involved in cloning and characterizing the biological activities of a number of glycosyltrasferases that perform the different steps of mucin type O-glycosylation. This project utilizes classical purification and cloning approaches and also takes advantage of the human genome project to clone related cDNAs for proteins of similar sequence to known enzymes and then characterize their function, tissue distribution, and patterns of expression in different disease conditions.

Molecular studies on MUC4 (NIH R01 CA29088)
I am a co-investigator on a project (Principal Investigator Surinder Batra) that is designed to further characterize the sequence, expression patterns and biological function of the MUC4 mucin gene which is overexpressed in cases of pancreatic cancer and is not expressed in adult human pancreas. There isevidence of alternative splicing of the MUC4 mRNA by pancreatic tumor cells as compared to other organ sites.

Analysis of Differential Gene Expression During Differentiation and Carcinogenesis by DNA Chip Technology (Nebraska Research Initiative)
We have constructed cDNA libraries from several unique sources, including normal fetal and adult pancreas of different ages ,tumors of different metastatic phenotype, and tumors of different differentiation states. We have identified and published results on a number of differentially expressed genes by using molecular genetic techniques (subtraction cloning, differential display, differential cDNA screening). These include a series of novel genes that are differentially expressed by pancreatic cells of different normal and disease conditions. We have funding (with Principal Investigator Angie Rizzino and Co-Investigators Rob Lewis, Jim Shull, and Ming Fong Lin) to further characterize the expression of these genes and to extend these studies to large sets of genes of known and unknown function by employing DNA arrays.

Back to Top

• SPECIALIZED LABORATORY/CLINICAL RESEARCH RESOURCES:

Dr. Hollingsworth has access to 2000 sq. ft. of laboratory space and 200 sq. ft. of office space in the Eppley Institute on the UNMC campus. Available core facilities include facilities for cell culture, mutagenesis, NMR and mass spectrometry, histology, animal care, electron microscopy, Storm phosphorimager, monoclonal antibody production, DNA sequencing, oligonucleotide production, capillary electrophoresis, 2-dimensional gel electrophoresis and analysis, and large scale production of recombinant products.

The P.I's lab area contains:  Centrifuges, pH meters, spectrophotometer, fluorescence and conventional microscopes, electrophoresis and isoelectric focusing equipment, laminar flow hoods, liquid scintillation and a gamma counter, CO2 incubators, an HPLC, gel electrophoresis equipment, bacterial incubators, superspeed centrifuges, and ultracentrifuge, a therymal cycler, and a hybridization oven.

Back to Top