July 2011


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"Diane St. Germain" <[log in to unmask]>
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Wed, 20 Jul 2011 09:49:09 -0400
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>>> *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.
>>>             ###