# Uncovering novel gene regulatory mechanisms underlying glucocorticoid response phenotypes through targeted mutagenesis of an essential transcription factor

> **NIH NIH K01** · DUKE UNIVERSITY · 2022 · $143,585

## Abstract

ABSTRACT
Hormone signaling and endocrine therapies are critical to human health. Genetic variation altering endocrine
responses contributes to the risk of developing endocrine disorders and metabolic diseases. A predominant
mechanism underlying altered endocrine responses involves coding variants in the transcription factors (TF)
regulating these signaling pathways. Characterizing the regulatory effects of TF coding variants en masse
remains challenging, thus limiting our understanding of how key endocrine pathways are controlled. To change
this paradigm, the objective of this proposal is to determine how pathogenic coding variants in the
glucocorticoid receptor (GR), a major and representative regulator of steroid hormone signaling, alter the
genomic response to glucocorticoids (GCs). There are thousands of known mutations in the GR, and likely
many more that have yet to be observed. For nearly all of those variants, the effects on GR activity are
unknown. Further, the prevalence of specific GR variants differs across populations and may contribute to the
diverse range of individual responses to pharmaceutical and physiological stimuli signaled though the GR. This
proposal directly addresses how variation (GR mutants) impacts phenotype (gene expression) by testing my
central hypothesis that genetic variation in the GR alters subsets of the genomic GC response by modifying
interactions with other TFs and co-factors. Guided by preliminary data, I will test my central hypothesis by
completing two Specific Aims. In Aim 1, I will simultaneously measure, in a single assay, the effects of 194
mutant GR genotypes on GC-responsive gene expression via a high-throughput reverse genetic screening
platform. I will test GR coding variants that are either pathogenic, preclude post-translational modification, or at
the extremes of positive or negative selection. Changes in the GC-responsive transcriptome will identify loss-
of-function, sub-pathological, and benign coding mutations thus yielding an empirical pathogenicity score for
each variant. In Aim 2, I will measure how specific clinically relevant GR mutations alter the genomic response
to GCs and ultimately perturb cellular function in three independent homozygous tagged mutant GR cell lines.
These Aims leverage my existing skill sets and domain expertise against training in genome editing, single-cell
technologies, and statistical analyses I require to achieve scientific independence. To aid my transition to
becoming an independent investigator, I have formed an interdisciplinary Advisory Committee composed of
leading scientists at my institute. This committee will advise on my research and professional progress,
including providing feedback on tenure-track job applications and grants. I will use my accomplishments under
this award to publish peer-reviewed manuscripts in high impact journals, secure a tenure track position at an
R1 institute, and obtain independent NIH funding. Completion of the research...

## Key facts

- **NIH application ID:** 10449610
- **Project number:** 1K01DK128388-01A1
- **Recipient organization:** DUKE UNIVERSITY
- **Principal Investigator:** Graham Johnson
- **Activity code:** K01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $143,585
- **Award type:** 1
- **Project period:** 2022-03-09 → 2022-09-01

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10449610

## Citation

> US National Institutes of Health, RePORTER application 10449610, Uncovering novel gene regulatory mechanisms underlying glucocorticoid response phenotypes through targeted mutagenesis of an essential transcription factor (1K01DK128388-01A1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10449610. Licensed CC0.

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