>>> *Thesis Defense Announcement
>>> To: The George Mason University Community*
>>>
>>> *Candidate: John R. Grimsley
>>> Program: Master of Science in Biology
>>> *
>>> *Date: Friday July 22, 2011
>>> Time: 9:30 a.m.
>>> Place: George Mason University, Prince William campus
>>> Bull Run Hall, Room 246
>>>
>>> Thesis Chair: Dr. Geraldine Grant
>>>
>>> Title: **"The Role of* *Krüppel Like Factor 4 in Idiopathic Pulmonary Fibrosis**"*
>>>
>>>
>>>
>>> A copy of the thesis is on reserve in the Johnson Center Library,
>>> Fairfax campus. The thesis will not be read at the meeting, but
>>> should be read in advance.
>>>
>>> All members of the George Mason University community are invited to
>>> attend.
>>>
>>>
>>> ABSTRACT:
>>>
>>> Idiopathic Pulmonary Fibrosis (IPF) is a fatal
>>> interstitial lung disease (ILD) with no known cause and
>>> characterized by a progressive build up fibrotic tissue
>>> in the diseased lung. Krüppel-Like Factor 4 (KLF4), a
>>> transcription factor with key roles in the cell cycle,
>>> cellular differentiation and development, was found to
>>> be over-expressed in primary IPF fibroblasts. In this
>>> study we sought to investigate the potential role of
>>> KLF4 in IPF by investigating the effect of its
>>> over-expression on fibroblast differentiation and
>>> proliferation status in the normal human pulmonary
>>> fibroblasts cell line, MRC5. In addition, we
>>> investigated the localization of KLF4 in vivo using
>>> normal and IPF tissue (LTRC). The in vivo localization
>>> of KLF4 in IPF and normal tissue was confirmed by
>>> double-Immunohistochemistry (IHC) in combination with
>>> either alpha-SMA or PCNA (Abcam). KLF4 over-expression
>>> in MRC5 cells was achieved using a doxycycline inducible
>>> lentiviral system (Addgene) and confirmed by
>>> quantitative real time PCR (Q-RTPCR) and western blot.
>>> The effect of KLF4 over-expression on markers of
>>> proliferation and activation was monitored by Q-RTPCR.
>>> Localization of KLF4 in the IPF lung was observed on the
>>> perimeter of the fibroblastic foci and in the parenchyma
>>> in areas of advanced fibrosis, however, absent with in
>>> the fibrotic foci. Co-localization of KLF4 and PCNA was
>>> observed, however, alpha-SMA and KLF4 were not observed
>>> to co-localize. Following KLF4 induction in vitro a
>>> 6-fold increase in KLF4 mRNA and a 3-fold increase in
>>> protein were observed. A decrease in alpha-SMA
>>> expression (2-fold) at both the gene and protein level
>>> was detected in addition to an increase in collagen 1A1
>>> (3-fold) mRNA. A decrease in all proliferation markers
>>> selected: PCNA, cyclin D1 and p21was observed, in
>>> addition to p53. The in vivo distribution of KLF4,
>>> suggests may be involved in the advancement of fibrosis
>>> and expansion of the foci. In vitro over-expression
>>> studies in MRC5 cells revealed a profoundly negative
>>> affected on the cell cycle which appears to be in
>>> contrast of IPF, however, in keeping with KLF4's dual
>>> roles of tumor suppressor and tumor promoter. Of
>>> particular interest was KLF4 over-expression resulted in
>>> an increase in collagen production, the main ECM
>>> component of IPF. KLF4 operated in a strongly context
>>> dependant manner, and IPF is a particularly context
>>> driven disease where the ECM acts almost as another
>>> pathogenic entity. Therefore, while direct comparison
>>> between in vivo IPF fibroblasts and in vitro MRC5 cells
>>> cannot be made, these data indicates a potential role
>>> for KLF4 in the pathogenesis of IPF. Further
>>> characterizations of this role through experimentation
>>> in primary adult IPF fibroblasts are warranted.
>>>
>>> ###
>>>
|
