> *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: 11:00 a.m.
> Place: George Mason University, Prince William campus
> Bull Run Hall, Room 257
>
> 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.
>
> ###
>
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