# Bacterial and Molecular Determinants of Mycobacterial Impermeability

> **NIH NIH R01** · UNIVERSITY OF VIRGINIA · 2024 · $716,926

## Abstract

Project Summary
The typical course of treatment for uncomplicated Mycobacterium tuberculosis infection comprises
four antibiotics and lasts for at least six months. The impermeability of the multi-layered M.
tuberculosis cell envelope has long been linked to the organism’s intrinsically-poor drug susceptibility.
While the outer ‘myco’ membrane is hypothesized to be the primary barrier to accessing antibiotic
targets in the peptidoglycan or cytoplasm, other facets of the envelope likely contribute. Moreover,
compared to the model organism Escherichia coli and to mammalian cells, the field has only a
rudimentary understanding of the kinds of compounds that can permeate M. tuberculosis. Based on
published and preliminary data, the PIs hypothesize that M. tuberculosis impermeability is a
consequence of envelope composition as well as the target location and chemical structure of the
compound. A major hurdle to testing this hypothesis is the lack of high-throughput tools for identifying
bacterial and molecular factors that control compound permeation across envelope layers. In this
proposal, The PIs develop and deploy two complementary methods for defining the bacterial and
molecular determinants of M. tuberculosis impermeability. They will first test the relative importance of
various M. tuberculosis factors in molecule gate-keeping. Next, they will globally determine the M.
tuberculosis genes that contribute to mycomembrane impermeability. Finally, they will
comprehensively determine the structural motifs associated with M. tuberculosis mycomembrane
permeation or lack thereof. Successful completion of these aims will lay the foundation for medicinal
chemistry efforts to improve M. tuberculosis uptake of both existing drugs and newly-discovered
compounds. The methods that the PIs use to achieve these aims are easily ported to species for
which envelope permeability is also treatment-limiting, e.g., non-tuberculous mycobacteria (NTMs)
and Gram-negatives.

## Key facts

- **NIH application ID:** 10898003
- **Project number:** 5R01AI179080-02
- **Recipient organization:** UNIVERSITY OF VIRGINIA
- **Principal Investigator:** Marcos M. Pires
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $716,926
- **Award type:** 5
- **Project period:** 2023-08-02 → 2028-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10898003

## Citation

> US National Institutes of Health, RePORTER application 10898003, Bacterial and Molecular Determinants of Mycobacterial Impermeability (5R01AI179080-02). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10898003. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*