3043 Induction of Kruppel-like Transcription Factor (KLF4&5) By Baker's Yeast Mannan in Human Bronchial Epithelial and Smooth Muscle Cells

Friday, 16 October 2015
Hall D1 Foyer (Floor 3) (Coex Convention Center)

Dukhee/Betty Lew, MD , Pediatrics, University of Tennessee/Le Bonheur Children's Hospital's, Memphis, TN

Kim/S. Lemessurier, PhD , Pediatrics, University of Tennessee/Le Bonheur Children's Hospital's, Memphis, TN

Joseph/a Moore, BS , Pediatrics, University of Tennessee/Le Bonheur Children's Hospital's, Memphis, TN

Jeoung-Eun Park, PhD , Pediatrics, University of Tennessee/Le Bonheur Children's Hospital's, Memphis, TN

Ae-Kyung Yi, PhD , Pediatrics, University of Tennessee/Le Bonheur Children's Hospital's, Memphis, TN

Chi/Young Song, PhD , Pharmacology, University of Tennessee/Le Bonheur Children's Hospital's, Memphis, TN

Kafait/U Malik, PhD , Pharmacology, University of Tennessee/Le Bonheur Children's Hospital's, Memphis, TN

Dukhee/Betty Lew, MD , Pediatrics, University of Tennessee/Le Bonheur Children's Hospital's, Memphis, TN

Rationale: Mannan derived from Saccharomyces cerevisiae(SC-MN) modulates allergic asthma pathogenesis in a mouse model. The purpose of this study is to explore downstream transcription factor(s) involved in SC-MN’s beneficial effects in asthma. The KLFs are important transcription factors in epithelial survival, and modulate epithelial mesenchymal transition and smooth muscle proliferation. As KLF4&5 are highly expressed in lung, we hypothesize that SC-MN can induce KLFs in lung cells.

Methods: Primary normal human epithelial cells (NHBEC) and bronchial smooth muscle cells were incubated with SC-MN and examined for KLF4, KLF5, p38MAPK levels and phosphorylation over a time course by Western Blot (WB). Normal human bronchial smooth muscle cells were analyzed for SM22alpha levels by WB and alpha-isoactin by indirect immunofluorescent staining.

Results: Following exposure to SC-MN, protein levels of KLF4 and KLF5 increased in both NHBEC and NHBSM cells over the subsequent 2-18 h. SC-MN–treated bronchial smooth muscle cells, but not in NHBEC, showed biphasic activation of p38MAPK (5-120 min and 8 h) that is known to lead to KLF phosphorylation in vascular smooth muscle cells. In addition, SC-MN increased smoooth muscle specific alpha-isoactin and SM22alpha at 24 hrs, consistent with a phenotype change.

Conclusions: SC-MN can induce KLF4 or KLF5, transcription factors that are important in epithelial survival and regulation of smooth muscle proliferation.