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A7630 - Antisense Oligonucleotide Mediated Reduction of SPDEF Regulates Goblet Cell Formation in Multiple Murine Pulmonary Disease Models
Author Block: J. R. Crosby, C. Zhao, D. Bai, M. Zhang, B. P. Monia, S. Guo; Drug Discovery, IONIS Pharmaceuticals Inc, Carlsbad, CA, United States.
RATIONALE: Many chronic pulmonary diseases, including cystic fibrosis (CF), asthma and chronic obstructive pulmonary disease (COPD), are characterized by excessive mucus production and increased numbers of goblet cells. SAM pointed domain-containing Ets transcription factor (SPDEF) is expressed in airway epithelial cells of the conducting airway and regulates goblet cell differentiation and mucus production. Overexpression of SPDEF in mouse lung causes goblet cell metaplasia while deletion of this gene resulted in the absence of goblet cells in the conducting airway epithelium. We have evaluated an antisense strategy for the treatment of several murine disease models in lung with the goal of preventing mucus overproduction by inhibiting the transcription factor SPDEF.
METHODS: Here we assessed the effectiveness of mouse SPDEF specific antisense oligonucleotides (ASOs) in a β-ENaC mouse model of CF, a steroid resistant severe asthma (SA) model and a wood smoke (COPD) model. The SA model is induced by nasal delivery of house dust mite (HDM) and cyclic-di-GMP administered over the course of 28 days. The wood smoke COPD study was conducted at the Lovelace Research Institute. Mice were exposed to wood smoke or filtered control air in a whole-body exposure system six hours a day, five days a week for four weeks. ASOs were delivered directly to the lung by inhalation or orotracheal delivery and mRNA levels were determined after density gradient fractionation of lungs and RT-PCR. Mucus production and Airway hyperresponsiveness (AHR) was determined.
RESULTS: SPDEF ASO delivery to the lungs of mice in multiple models of pulmonary disease resulted in effective reduction of target mRNA. In the β-ENaC CF model with established lung disease, SPDEF ASOs reduced goblet cell metaplasia and mucus production, AHR and neutrophil recruitment to the airways. In the severe asthma model, SPDEF ASOs were also effective at reducing mucus genes, PAS-positive mucus and AHR. In the COPD model, the wood smoke but not filtered air induced SPDEF mRNA and other mucus genes which were all reduced by SPDEF ASO inhalation. In addition, AHR was improved.
CONCLUSIONS: Targeted knockdown of SPDEF by ASO reduced mucus production and prevented or reversed goblet cell transdifferentiation in CF, SA and COPD mouse models. These data demonstrate that ASO inhibition of SPDEF in airway could be an effective approach for the treatment of multiple pulmonary diseases with excess mucus.