> *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. > > ### >