Jenny Thiele
Date & Time: Thursday, March 25, 2010, 10:00 am
Location: Room
2027 at The Centre for Molecular Medicine and Therapeutics, 950 West 28th
Avenue, Vancouver, BC.
“Generation and characterization of embryonic stem cell
lines derived from the
YAC128 mouse model of Huntington disease”
Jenny Thiele MSc Programme (pdf)
EXAMINING COMMITTEE
Chair: Dr. Pamela Hoodless (Medical Genetics)
Supervisory Committee: Dr. Blair Leavitt, Research
Supervisor (Medical Genetics) & Dr. Fabio Rossi (Medical Genetics)
University Examiner: Dr. Dan Goldowitz (Medical Genetics)
SUPERVISORY COMMITTEE
Dr. Blair Leavitt, Research Supervisor (Medical Genetics), Dr.
Fabio Rossi (Medical Genetics), & Dr. Elizabeth M. Simpson (Medical Genetics)
ABSTRACT
Huntington Disease (HD) is an autosomal dominant
neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in
the Huntingtin gene. Patients typically present in mid-life with progressive
motor dysfunction, cognitive deficits, and neuropsychiatric abnormalities.
Recently, researchers have provided evidence that HD is associated with
significant pathology in peripheral tissues as well. At the current time no
effective treatment has been proven to alter or cure progression of HD which
leads to complete loss of independence and eventual death an average of 20
years after disease onset.
The ability to model Huntington disease in animals has
enabled studies which have provided new insights into the mechanisms of HD
pathogenesis. However, the development of simple cell culture-based systems
will be useful to accelerate our research efforts into the basic underlying
pathogenic pathways of HD and will allow dissection of cellular interactions
and the identification of novel targets for intervention that offer the
greatest hope of a cure. The YAC mouse model of HD expresses full-length human
Huntingtin with either 18 polyglutamines (YAC18) or 128 polyglutamines
(YAC128), and develops age-dependent cognitive deficits, motor dysfunction, and
selective striatal neurodegeneration similar to that seen in human HD patients.
I have generated novel embryonic stem (ES) cell lines from wild-type, YAC18 and
YAC128 mice on two genetic backgrounds. These cell lines have been cultured
under defined conditions over long periods of time, and express characteristic
markers of pluripotency, such as alkaline phosphatase, Oct-4 and Nanog. Neurons
and macrophages derived from these novel cell lines using established in vitro
protocols have been characterized via immunocytochemistry and challenged in
functional assays.
To confirm results attained from functional assays in our
ES-derived macrophages, I examined primary macrophages and microglia cultures derived
from the YAC mice and determined the functional response of these cells to
endotoxin stimulation. Primary cell cultures isolated from YAC128 mice produced
significantly more IL-6 than wild-type cultures. In comparison, with the same
endotoxin stimulation, YAC18 primary macrophages and microglia responded with
similar levels of IL-6 release as cultures of wild-type cells, suggesting that
the over-activity in the YAC128 cytokine response is caused by the mutant
Huntingtin trans-gene.
PUBLICATIONS
Thiele J and Van Raamsdonk JM: Gene discovery in
methylmalonic aciduria and homocystinuria. Clinical Genetics 69(5):402-3 (2006).
AWARDS
Research methodology training grant - Child & Family
Research Institute, University of British Columbia for $3,000 (2009).
PRESENTATIONS
The Centre for Molecular Medicine and Therapeutics TGIF
Series. October 2008, Vancouver (Canada) Seminar Presentation – “New insights
into HD pathogenesis: Altered cellular inflammatory responses”.
World Congress on Huntington’s Disease. September 2007,
Dresden (Germany)
Poster Presentation – “ES cells derived from YAC128
transgenic mice; a novel in vitro model for Huntington Disease”.
The Child & Family Research Institute Student Research
Forum . June 2007, Vancouver (Canada). Poster Presentation – “ES cells derived
from YAC128 transgenic mice; a novel in vitro model for Huntington Disease”.
The Centre for Molecular Medicine and Therapeutics TGIF
Series. November 2006, Vancouver (Canada). Seminar Presentation – “ES cell
differentiation - A tale of two cell types”.
GRADUATE STUDIES
Field of Study: Huntington disease
COURSES:
MEDG 419: Human Cytogenetics, Dr. Carolyn Brown
MEDG 505: Genome Analysis, Dr. Philip Hieter
MEDG 520: Advances in Human Molecular Genetics, Dr. Matthew
Lorincz
MEDG 530: Human Genetics, Dr. Lorne Clarke
MEDG 545: Current Topics in Medical Genetics Research -
Molecular Mechanisms of Disease, Dr. Michael Kobor
MEDG 548A: Directed Studies, Dr. Blair Leavitt
Jenny Thiele MSc programme (pdf)