>>> *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. >>> >>> ### >>>