Current
Position
- Chair in Cancer
Research Northeastern Ontario Regional
Cancer Centre, Sudbury, Ontario
- Professor, Department
of Chemistry and Biochemistry, Laurentian
University, Sudbury, Ontario
- Assistant Professor,
Department of Medicine, Division of Oncology,
University of Ottawa, Ottawa, Ontario
- Assistant Professor,
Department of Biochemistry, Molecular
Biology and Immunology, University of
Ottawa, Ottawa, Ontario
Education
and Training
1991-1995 Postdoctoral
Research Fellowship in Medicine, Banting
and Best Department of Medical Research,
University of Toronto, Toronto, Ontario
1989-1991 Postdoctoral
Research Fellow in Medicine, Harvard Medical
School & Joslin Diabetes Center, Boston,
Massachusetts, USA
1989 Ph.D.York University
(Biochemistry), Toronto, Ontario
1983 B.Sc. University
of Guelph (Biochemistry), Guelph, Ontario
Awards
and Honours
2002
Government of Ontario, Premier’s Research
Excellence Award, Value: $150,000
1992-1995 Canadian Cystic Fibrosis Foundation
Postdoctoral Fellowship
1989-1991 Joslin Diabetes Center Training
Fellowship, Elliot P. Joslin Research Laboratory,
Harvard Medical School
1987-1988; 1988-1989 Ontario Graduate Scholarship,
Ministry of Colleges and Universities, Government
of Ontario
1984-1985; 1985-1986 Graduate Assistantships,
Dept. of Biology, York University, Toronto,
Ontario
Research
Funding
- Xanthus
Pharmaceuticals, Cambridge, MA
- YM
Biosciences Inc., Toronto, ON
- Ontario
Cancer Research Network
- Canada
Foundation for Innovation
- Ministry
of Science and Technology
- R.
Samuel McLaughlin Foundation
- Canadian
Institute of Health Research
- Ontario
Research and Development Challenge Fund
- Medical
Research Council of Canada
- Northern
Cancer Research Fund
- Canadian
Breast Cancer Foundation
Lab
Staff
Research Investigations
Protein
Kinase C Signaling and Multidrug Resistance
in Breast Cancer
There are two major research foci for my
research group. One relates to increasing
our understanding of how the activity of
the protein kinase C (PKC) family of kinases
is regulated in cells. Disregulation of
PKC isoenzymes is associated with many disease
states, including cancer and diabetes. Our
second research focus is "multidrug
resistance", where tumours exhibit
or acquire the ability to resist killing
by a wide spectrum of structurally unrelated
chemotherapy drugs. Our goals are to better
understand the various mechanisms responsible
for multidrug resistance and to improve
the effectiveness of chemotherapy in patients.
Current research projects include:
Research
Projects:
1- Multidrug Resistance in Breast
Tumour Cells
We have established a panel of isogenic
cell lines that have been selected for resistance
to various chemotherapy drugs used in the
treatment of human breast cancer. Since
the cell lines all originate from the same
source and were selected in an identical
manner, they should be invaluable in examining
the relationship between the expression
of specific genes or proteins in breast
tumour cells and drug resistance. Using
cDNA micro-array and other approaches, we
have identified a variety of genes, whose
expression correlates with resistance of
breast tumour cells to specific drugs in
vitro. In an upcoming clinical trial by
the National Cancer Institute of Canada,
we will examine the utility of these genes
as predictors of clinical response in patients
undergoing chemotherapy for locally advanced
breast cancer. We are also identifying agents
that can effectively kill drug-resistant
breast tumour cells. One such agent (calphostin
C) is capable of killing cell lines that
are > 4000-fold more resistant to drug
than wildtype cell lines.
2-Mapping of newly identified novel
PKC inhibitory sites within the PKC Regulatory
Domain
We have recently published evidence that
the activity of the protein kinase C alpha
(PKCa) catalytic domain can be potently
and specifically inhibited by its regulatory
(R) domain without requiring the "pseudosubstrate"
site within the PKC R domain. Since there
are no additional pseudosubstrate like sites
within this R domain, this observation challenges
the current exclusive role of pseudosubstrates
in the autoinhibition of protein kinases
and suggests that additional site(s) are
involved. We are currently localizing and
characterizing these additional PKC inhibitory
sequence(s) with the goal of better understanding
their mechanism of autoinhibition. Through
deletion analysis, we have now established
that one inhibitory region lies between
amino acids 36-177. Since this region lacks
the PKC, Ca++ and phosphatidylserine binding
sites, and since inhibition can occur in
a phorbol ester-independent manner, it appears
that the mechanism does not involve sequestration
of PKC activators/cofactors. Possible inhibition
of PKC catalytic activity by binding the
PKC substrate used in our assays has also
been ruled out. The additional sites within
the PKC R domain contributing to PKC autoinhibition
are being more finely localized by alanine
linker-scanning and site directed mutagenesis
approaches.
3-
Modulation of PKC function by the Cytoskeletal
Protein Calponin
Protein kinase C (PKC) is a family of phosphatidylserine-dependent
protein kinases which play important roles
in regulating a variety of cellular processes.
Aberrant regulation of PKC has been implicated
in the etiology of a variety of disease
states including cancer and the vascular
complications associated with diabetes.
The activity of "classical" PKCs
(a, b, g) is regulated by a variety of hormonal
stimuli which induce a translocation of
the enzyme to specific subcellular compartments
and an increase in PKC catalytic activity.
One mechanism for increasing PKC activity
involves the hormone receptor-coupled hydrolysis
of a membrane phosphatidid (PIP2) which
stimulates diacylglycerol (DAG) and Ca++
production resulting in elevated PKC activity.
We have recently identified an additional
mechanism for activating PKC. We have observed
that the cytoskeletal protein calponin can
also activate PKC activity without requiring
the hydrolysis of PIP2. The protein binds
to PKC directly and induces autophosphorylation
of the enzyme. Calponin also appears to
increase the amount of PKC bound to membranes.
We are currently localizing the calponin-binding
domain(s) within PKC and determining whether
calponin induces a change in conformation
for PKC which is different from that induced
by other PKC activators (DAG and Ca++).
Selected
Publications
Refereed
Papers
1.
Parissenti, A.M., Kim, S.A., Snihura, A.,
Colantonio, C.M. and Schimmer, B.P. (1996)
The regulatory domain of human protein kinase
C alpha dominantly inhibits protein kinase
C beta- regulated growth and morphology
in Saccharomyces cerevisiae. J. Cell. Physiol.
166, 609-617.
2.
Parissenti, A.M., Kirwan, A.F., Kim, S.A.,
Colantonio, C.M. and Schimmer, B.P. (1996).
Molecular strategies for the dominant inhibition
of protein kinase C. Endocrine Res. 22,
621-630.
3.
Parissenti, A.M., Kirwan, A.F., Colantonio,
C.M., Kim, S.A. and Schimmer, B.P. (1998).
Inhibitory properties of the regulatory
domains of human protein kinase C alpha
and mouse protein kinase C epsilon. J. Biol.
Chem. 273, 8940-8945.
4. Parissenti, A.M., Villeneuve, D., Kirwan-Rhude,
A.F. and Busch, D. (1999). A carbon source-dependent
regulation of cell growth by murine PKC
epsilon expression in Saccharomyces cerevisiae.
J. Cell. Physiol. 178, 216-226.
5.
Parissenti, A.M., Gannon, B.R., Villeneuve,
D.J., Kirwan-Rhude, A.F., Chadderton, A.
and Gluck, S. (1999). Lack of modulation
of MDR1 gene expression by dominant inhibition
of cAMP-dependent protein kinase in doxorubicin-resistant
MCF-7 breast cancer cells. Int. J. Cancer
82, 893-900.
6.
Chadderton, A., Villeneuve, D.J., Gluck,
S. Kirwan-Rhude, A.F., Gannon, B.R., and
Parissenti, A.M. (2000). Role of specific
apoptotic pathways in the restoration of
paclitaxel-induced apoptosis by valspodar
in doxorubicin-resistant MCF-7 breast cancer
cells. Breast Cancer Res. Treat. 59, 231-244.
7.
Leinweber, B., Parissenti, A.M., Kirwan-Rhude,
A.F., Leavis, P.C. and Morgan, K.G. (2000).
Regulation of PKC by the cytoskeletal protein
calponin. J. Biol. Chem. 275, 40329-40336.
8.
Bewick, M., Conlon, M., Gerard, S., Lee,
H., Parissenti, A.M., Zhang, L., Gluck,
S. and Lafrenie, R.M. (2000). HER-2 expression
is a prognostic factor in patients with
metastatic breast cancer treated with a
combination of high-dose cyclophosphamide,
mitoxantrone, paclitaxel, and autologous
blood stem cell support. Bone Marrow Transplantation
27, 847-853.
9.
Bewick, M., Conlon, M., Parissenti, A.M.,
Lee, H., Zhang, L., Gluck, S., and Lafrenie,
R.M. (2001). Soluble Fas (CD95) is a prognostic
factor in patients with metastatic breast
cancer undergoing high dose chemotherapy
and autologous stem cell transplantation
J. Hematother. Stem Cell Res. 10, 759-768.
10.
Kirwan, A. F., Bibby, A. C., Mvilongo, T.,
Riedel, H., Burke, T., Millis, S. Z. and
Parissenti, A. M. (2003) Inhibition of PKC
catalytic activity by additional regions
within the human PKC alpha regulatory domain
lying outside of the pseudosubstrate sequence.
Biochem. J. 373, 571-581.
11.
Riedel, H. and Parissenti, A.M. (2003) The
use of Saccharomyces cerevisiae to study
structure/function relationships for mammalian
protein kinase C isoforms and their modulators.
Methods in Molecular Biology 233: 491-516.
12.
Guo, B., Hembruff, S. L., Villeneuve, D.
J., Kirwan, A. F. and Parissenti, A. M.
(2003) Potent killing of paclitaxel- and
doxorubicin-resistant breast cancer cells
by calphostin C accompanied by cytoplasmic
vacuolization. Breast Cancer Res. Treat.
82 (2), 125-141.
13.
Guo, B., Villeneuve, D. J., Kirwan, A. F.,
Hembruff, S. L., Blais, D. E., Bonin, M.
and Parissenti, A. M. (2004) Cross resistance
studies of isogenic drug-resistant breast
tumour cell lines support clinical evidence
suggesting that sensitivity to paclitaxel
may be strongly compromised by prior doxorubicin
exposure. Breast Cancer Res. Treat. 85:31-51.
14.
Parissenti, A.M. and Villeneuve, D.J. (2004).
Invited Review: DNA microarrays and their
use in investigating the pharmacogenomics
of drug response. Current Topics in Medicinal
Chemistry 4:1329-1345.
Manuscripts in Preparation
1.
Villeneuve, D.J., Hembruff, S.L., and Parissenti,
A.M. (2005) cDNA Microarray Analysis of
Isogenic Paclitaxel- and Doxorubicin-Resistant
Breast Tumor Cell Lines Reveals Distinct
Drug-Specific Genetic Signatures of Resistance
(submitted).
2.
Hembruff, S.L., Villeneuve, D.J. and Parissenti,
A.M. (2005) Optimization of real-time PCR
methods for the confirmation of cDNA microarray
data. (submitted).
3.
Sprowl, J., Hembruff, S.L., Villeneuve,
D.J., and Parissenti, A.M. (2005) Regulation
of Cell Wall Synthesis-Related Genes Accompanies
Growth Arrest by Bovine Protein Kinase C
Expression in Saccharomyces Cerevisiae.
(manuscript in preparation).
4.
Guo, B., Reed, K. and Parissenti, A.M. (2005)
Scanning mutagenesis studies reveal multiple
distinct regions within human protein kinase
C alpha regulatory domain important for
phorbol ester – dependent activation
of enzyme”. (manuscript in preparation).
5.
Kozarova, A., Reed, K., Lafrenie, R., Hudson,
J.W. and Parissenti, A.M. (2005) Interaction
between the Regulatory and Catalytic Domains
of Human PKC? involving Sites Lying Outside
of the V1 Region”. (manuscript in
preparation)
Book Chapters
1.
Riedel, H. and Parissenti, A.M. (2003) The
use of Saccharomyces cerevisiae to study
structure/function relationships for mammalian
protein kinase C isoforms and their modulators.
Methods in Molecular Biology 233, 491-516.
Abstracts
1.
Parissenti, A.M., Kirwan, A., Colantonio,
C.M., Kim, S.A., Villeneuve, D., Snihura,
A. and Schimmer, B.P. (1995). Dominant inhibition
of protein kinase C isoforms by expression
of protein kinase C regulatory domains in
Saccharomyces cerevisiae. Poster presented
at the Ontario Cancer Treatment and Research
Foundation Biennial Lake Couchiching Conference,
Orillia, Ontario, October 16-18.
2.
Parissenti, A.M., Gannon, B.R., Chadderton,
A. and Gluck, S. (1997). A genetic approach
for studying the role of cAMP-dependent
protein kinase in multidrug resistance in
human breast cancer cells. Seminar presented
at the "Symposium on Tumour Cell Signalling
and Cancer Therapy" conference in Hamilton,
May 22-23.
3.
Parissenti, A.M., Gannon, B.R., Chadderton,
A. and Gluck, S. (1997). Lack of modulation
of MDR1 gene expression by dominant inhibition
of cyclic AMP-dependent protein kinase in
adriamycin-resistant human MCF-7 breast
cancer cells. Seminar presented at the 17th
International Congress of Biochemistry and
Molecular Biology in San Francisco August
24 29.
4. Deaden, T., Chadderton, A., Yan, B.,
Bewick, M., Rhude, A., Parissenti, A. and
Glück, S. (1998). Probenecid as a potential
modulator of drug resistance in the MRP
overexpressing breast cancer cell line MCF7-VP.
Proc. of ASCO 239a.
5. Kirwan-Rhude, A.F. and Parissenti, A.M.
(1998). Localization of a novel protein
kinase C (PKC) inhibitory site within the
human PKC alpha regulatory domain. Invited
speaker to the “Protein Kinase C and
Cellular Function” Fall Symposium
sponsored by the American Society of Biochemistry
and Molecular Biology, Granlibakken, Lake
Tahoe, California, October 9-12
6.
Chadderton, A., Villeneuve, D.J., Kirwan-Rhude,
A.F., Gluck, S., Gannon, B.R., Blais, D.E.
and Parissenti, A.M. (1999). Restoration
of paclitaxel-induced chemosensitivity and
apoptosis in adriamycin-resistant MCF-7
breast cancer cells by inhibition of P-glycoprotein
using SDZ-PSC833. Poster presented at the
American Association for Cancer Research
Annual Meeting in Philadelphia, Pennsylvania,
April 10-14.
7.
Chadderton, A., Villeneuve, D.J., Gluck,
S., Kirwan-Rhude, A.F., Gannon, B.R., Blais,
D.E. and Parissenti, A.M. (1999). Restoration
of paclitaxel-induced apoptosis by valspodar
in doxorubicin-resistant breast tumour cells.
Poster presented at the biennial Cancer
Care Ontario Lake Couchiching Conference
in Orillia, Ontario, November 8-11.
8.
Leinweber, B., Parissenti, A.M. and Morgan,
K.G. (1999). Calponin binds the regulatory
domain of protein kinase C epsilon and activates
it kinase activity. Presented at the American
Society for Cell Biology meeting in Washington,
D.C. December 11-15.
9.
Parissenti, A. (2000). Breast cancer: Multidrug
resistance and prognostic indicators for
survival in patients undergoing high dose
chemotherapy. Seminar presented at The Toronto
Sunnybrook Cancer Centre, May 11, 2000.
Host: Kathy Pritchard, MD., Head, Clinical
Trials Division
10.
Parissenti, A. (2000). Protein kinase C
regulatory domains: Structure, function,
and utility as isoform-specific inhibitors
of protein kinase C function. Research seminar
presented at the Boston Biomedical Research
Institute, Boston, MA June 7, 2000. Host:
Kathy Morgan, Ph.D., Director
11.
Parissenti, A. (2001). Identification by
cDNA Microarray Analysis of Novel Genes
Correlating With Resistance to Specific
Drugs in Breast Cancer: Can these genes
predict drug responsiveness in patients?
Seminar presented at Breast Cancer Rounds
at the Toronto Sunnybrook Regional Cancer
Centre, Toronto, Ontario July 19, 2001 Host:
Kathy Pritchard, M.D. and Maureen Trudeau,
M.D.
12.
Parissenti, A. (2001). Identification and
localization of novel regions within the
protein kinase C alpha regulatory domain
responsible for protein kinase C autoinhibition,
activation, and targeting. Invited speaker,
Department of Biology Seminar Series, York
University, Toronto
13.
Parissenti, A. (2001) Pharmacogenomics of
Resistance to Anthracyclines and Taxanes
in Human Breast Cancer. Invited speaker
at the biennial Cancer Care Ontario Lake
Couchiching Conference in Orillia, Ontario,
November 8-11.
14.
Parissenti, A. (2004) The Use of Microarray
Technologies in Cancer Diagnosis, Prognosis
Prediction, and Treatment. Invited speaker
at the Society of Northern Ontario Pathologists,
Sudbury, Ontario, January 22.
15.
Parissenti, A. (2004) Cross-Resistance Studies
In Vitro Support Recent Clinical Evidence
that Sensitivity to Paclitaxel is compromised
by Prior Doxorubicin Exposure. Invited speaker
at the 1st ISC International Conference
on Cancer Therapeutics, Molecular Targets,
Pharmacology and Clinical Applications,
Palazzo dei Congress, Florence, Italy, February
19-21.
16. Parissenti, A. (2005) Genomic Signatures
of Resistance to Anthracyclines and Taxanes
in Breast Cancer. Invited speaker at the
National Cancer Institute of Canada, Clinical
Trials Group Neoadjuvant Therapy Workshop,
Toronto, Ontario, February 21.
17.
Parissenti, A. (2005) Genomic Signatures
of Resistance to Anthracyclines and Taxanes
in Breast Cancer. Invited speaker at the
Princess Margaret Hospital, Toronto, February
22.
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