# Tissue-specific regulation of a gene essential for airway epithelial function

> **NIH NIH R01** · CASE WESTERN RESERVE UNIVERSITY · 2020 · $395,708

## Abstract

Summary
Significant progress has been made towards new therapies for Cystic Fibrosis (CF). This devastating inherited
disease is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene,
which encodes a multifunctional protein (CFTR) with a pivotal role in regulating anion transport across cell
membranes. CFTR is particularly well characterized in airway, pancreas, intestine and male genital duct
epithelia. Current pharmacological approaches to restore normal activity to defective CFTR focus on correcting
cellular trafficking of misfolding mutants and on potentiating activity of ion channels with faulty conductance.
For both classes of mutation increasing CFTR gene expression and hence the amount of CFTR protein
substrate, would likely enhance in vivo drug efficacy. Moreover, existing therapeutic goals will not benefit the
~15% of patients lacking sufficient functional CFTR due to mutations that disrupt gene expression. A detailed
understanding of the normal transcriptional mechanisms controlling the gene is a prerequisite for successful
approaches to modulate CFTR expression. In the previous R01 funding period we achieved our goal to
elucidate the tissue-specific control pathways of the CFTR gene in airway and intestinal epithelial cells. The
CFTR locus lies within a topologically associating domain (TAD) established by CCCTC-binding factor (CTCF)
insulator elements. Within this environment, distal cis-acting enhancers are brought into close association with
the gene promoter by a looping mechanism that is stabilized by the cohesin complex. The enhancers are cell-
type specific and associate with different activating or repressing transcription factors (TFs). Building on our
greatly improved understanding of CFTR regulation, we will pursue three specific aims addressing the
overarching hypothesis that targeted modulation of CFTR gene expression will increase functional CFTR
protein at the cell surface. Increasing transcript levels, alone or in combination with pharmacological
approaches, will alleviate disease phenotype. Further, we hypothesize that an enhanced understanding of the
cis-elements and interacting factors coordinating cell-type specific gene expression will reveal new ways to
augment CFTR transcription. In the first aim we will determine how cell-specific cis-regulatory elements
coordinate expression of the endogenous CFTR locus. Experiments will combine CRISPR/Cas9 targeting of
these elements with gene expression assays and analysis of locus architecture. In the second aim we will
decipher the transcriptional network that regulates CFTR expression in primary airway epithelial cells.
Candidate TFs will be examined by genome-wide methods to reveal their impact on the locus. In the third aim
we will identify and characterize pathways that activate CFTR gene expression to enhance CFTR mRNA and
functional CFTR protein expression. These results will enable novel approaches to increase CFTR in the
airway ...

## Key facts

- **NIH application ID:** 9903422
- **Project number:** 5R01HL094585-08
- **Recipient organization:** CASE WESTERN RESERVE UNIVERSITY
- **Principal Investigator:** Ann Harris
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $395,708
- **Award type:** 5
- **Project period:** 2009-01-01 → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9903422

## Citation

> US National Institutes of Health, RePORTER application 9903422, Tissue-specific regulation of a gene essential for airway epithelial function (5R01HL094585-08). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/9903422. Licensed CC0.

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