Antibiotic persistence remains one of the most challenging barriers to effective clearance of chronic bacterial infections. Despite the intervenin decades since its original discovery, we still lack a basic understanding of how molecular networks implement the defining features of persistence: phenotypic heterogeneity and tolerance to lethal levels of antibiotic exposure. Our work during the previous funding period, and recent observations by other groups, present a much more complex picture of persistence than previously thought. Building on our preliminary results, we propose to apply an integrated experimental/computational approach in order to achieve a new systems-level understanding of persistence.
We aim to accomplish this by discovering the most comprehensive set of genetic loci involved, mapping out the genetic network context in which these loci function and to reverse engineer the regulatory networks that reside at the core of persistence. These analyses will be aided by a systematic search for robust markers of the persistence cellular state. These markers will then be used to FACS-isolate sub-populations of cells exhibiting extremes of persistence. Each of these sub-populations will then be interrogated for the most comprehensive set of internal molecular states and regulatory interactions, utilizing a combination of existing methods and genomic technologies recently developed by our group. These include: transcriptomics, genome-wide in vivo DNA-protein interactions, transcriptome-wide in vivo RNA-protein interactions, RNA-RNA interactions, and ribosome-RNA interactions. The massive scale of these observations will reveal the most comprehensive and unbiased global view of the persistence cellular state, to date. Through computational analysis and integration of these observations we aim to learn compact graphical models of causal regulatory interactions that modulate persistence. These analyses will utilize powerful algorithms developed by our group, including information-theoretic pathway analysis, cis-regulatory analysis of linear (DNA/RNA) and structural RNA regulatory elements, along with Bayesian evidence integration. Beyond achieving a systems-level understanding of persistence, the composition and architecture of these molecular networks will serve as a knowledge-scaffold for development of novel strategies for eradicating chronic bacterial infections.

Public Health Relevance

The proposed research is a systems biology approach to characterize the molecular basis of antibiotic persistence. As persistence is central to clinical resistance of a variety of human infections, these studies will be foundational in development of future therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI077562-09
Application #
9485830
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Baqar, Shahida
Project Start
2010-07-01
Project End
2020-05-31
Budget Start
2018-06-01
Budget End
2019-05-31
Support Year
9
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Biochemistry
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
Khare, Anupama; Tavazoie, Saeed (2015) Multifactorial Competition and Resistance in a Two-Species Bacterial System. PLoS Genet 11:e1005715
Freddolino, Peter L; Goodarzi, Hani; Tavazoie, Saeed (2014) Revealing the genetic basis of natural bacterial phenotypic divergence. J Bacteriol 196:825-39
Hottes, Alison K; Freddolino, Peter L; Khare, Anupama et al. (2013) Bacterial adaptation through loss of function. PLoS Genet 9:e1003617
Freddolino, Peter L; Tavazoie, Saeed (2012) Beyond homeostasis: a predictive-dynamic framework for understanding cellular behavior. Annu Rev Cell Dev Biol 28:363-84
Freddolino, Peter L; Goodarzi, Hani; Tavazoie, Saeed (2012) Fitness landscape transformation through a single amino acid change in the rho terminator. PLoS Genet 8:e1002744
Freddolino, Peter L; Amini, Sasan; Tavazoie, Saeed (2012) Newly identified genetic variations in common Escherichia coli MG1655 stock cultures. J Bacteriol 194:303-6
Freddolino, Peter L; Tavazoie, Saeed (2012) The dawn of virtual cell biology. Cell 150:248-50
Girgis, Hany S; Harris, Kendra; Tavazoie, Saeed (2012) Large mutational target size for rapid emergence of bacterial persistence. Proc Natl Acad Sci U S A 109:12740-5
Khan, Zia; Amini, Sasan; Bloom, Joshua S et al. (2011) Accurate proteome-wide protein quantification from high-resolution 15N mass spectra. Genome Biol 12:R122
Amini, Sasan; Tavazoie, Saeed (2011) Antibiotics and the post-genome revolution. Curr Opin Microbiol 14:513-8

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