Media release

From: Macquarie University

Work is underway into how science can stop the superbug A. baumanniii after research exposes a weak link in the deadly but poorly understood pathogen.

The deadly hospital pathogen Acinetobacter baumannii can live for a year on a hospital wall without food and water. Then, when it infects a vulnerable patient, it resists antibiotics as well as the body's built-in infection-fighting response. The World Health Organization (WHO) recognises it as one of the three top pathogens in critical need of new antibiotic therapies.

Now, an international team, led by Macquarie University researchers Dr. Ram Maharjan and Associate Professor Amy Cain, have discovered how the superbug can survive harsh environments and then rebound, causing deadly infections. They have found a single protein that acts as a master regulator. When the protein is damaged, the bug loses its superpowers allowing it to be controlled, in a lab setting. The research is published this month in Nucleic Acids Research.

“We hope that our paper will encourage researchers worldwide to refocus on developing drugs to fight this superbug, which is spreading through the world’s hospitals, and killing already vulnerable people in intensive care units and other high-risk areas,” says Associate Professor Cain, the senior author on the paper.

There are six superbugs that scare global health officials. E. coli, Klebsiella pneumoniae and other gram-negative bacteria have common pathways that give them antibiotic resistance. A. baumannii is different. It’s particularly tough, and it’s one of the most resistant pathogens we encounter. Strangely, we don’t know much about how it infects us.

“In the lab we can see this pathogen is very tough. Other researchers have shown that you can desiccate the bug for a year and when they added water, it was still able to infect mice,” says Associate Professor Cain.

“The problem had been that A. baumannii is relatively new on the scene, emerging as a problem in hospitals in the 1980s. And it’s hard to genetically manipulate with the existing molecular biology toolkit. It usually only infects sick people, but it is very resistant to antibiotics making it incredibly hard to treat and difficult to safely research. So, we don’t know much about it. We don’t know where it came from, nor how it became so resistant and resilient. Now, thanks to this paper, we know how it deals with stress.”

Amy and her colleagues realised about five years ago that they could make a difference by trying to understand the underlying biology of A. baumannii. That led to a major investment by Macquarie University in the research, in biocontainment laboratories for staff safety, and in an ethical animal model using moth caterpillars. The research effort has been strongly supported by the Australian Council and the National Health and Medical Research Council.

During infection our cells fight back by either flooding or starving bacteria of essential metals such as copper and zinc. A. baumannii has strong drug pumps that push antibiotics, metals and other threats out of the cell.

“By studying how this bug deals with infection stresses, we’ve found an important uncharacterised regulatory protein (DksA). When we disrupt this protein, it leads to changes in about 20 per cent of the bug’s genome and breaks its pumping system,” says Dr Ram Maharjan, a Macquarie University researcher and first author on the paper.

Not only does this protein control stress response, but it also controls virulence. A. baumannii usually spreads in blood but our disruption also caused it to be completely undetected in the blood of both Galleria mellonella and mice. It also becomes super sticky and harmlessly sticks to organs.

This has been a massive global research effort over the past five years, working with colleagues at Flinders University, Monash University, University of Cambridge, University of Wurzburg.

The new paper builds on a discovery earlier this year also led by Macquarie researchers that showed that K. pneumoniae and A. baumannii work together to avoid antibiotics.

About Amy Cain

Associate Professor Amy Cain is an Australian Research Council Future Fellow. Following a PhD at the University of Sydney she has worked in a range of institutes and pharma companies. She joined Macquarie University about five years ago, to focus on antimicrobial resistance, recognising that the solutions to this wicked problem would be found in fundamental research into the life course of these pathogens.

About the ethical animal model

The Macquarie researchers are now working with caterpillars as an ethical animal model to screen for potential drugs. The larvae of the greater wax moth, Galleria mellonella usually live in beehives in the tropics where they largely live on beeswax. So, the caterpillars grow well at human body temperature which makes them ideal to study bacteria that grow in the body. The Macquarie Galleria Research Facility is the first of its kind in Australia.

Read about Macquarie Univeristy’s Galleria Facility, using caterpillars as an ethical animal model at https://www.mq.edu.au/faculty-of-science-and-engineering/departments-and-schools/applied-biosciences/our-facilities/galleria-facility

https://doi.org/10.1093/nar/gkad341

Abstract

DksA is a conserved master regulator of stress response in Acinetobacter baumannii

Ram P. Maharjan1, Geraldine J. Sullivan1, Felise G. Adams2, Bhumika S. Shah1, Jane Hawkey3, Natasha Delgado1, Lucie Semenec1, Hue Dinh1, Liping Li1, Francesca L. Short4, Julian Parkhill5, Ian T. Paulsen1, Lars Barquist 6,7, Bart A. Eijkelkamp2 and Amy K. Cain 1,*

1 ARC Centre of Excellence in Synthetic Biology, School of Natural Sciences, Macquarie University, Sydney, NSW 2109, Australia

2 College of Science and Engineering, Flinders University, Bedford Park, SA 5042, Australia

3 Department of Infectious Diseases, Central Clinical School, Monash University, Victoria, Australia, 4 Department of Microbiology, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, Australia

5 Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge CB3 0ES, UK

6 Helmholtz Institute for RNA-based Infection Research (HIRI), Helmholtz Centre for Infection Research (HZI), 97080 Würzburg, Germany

7 Faculty of Medicine, University of Würzburg, 97080 Würzburg, Germany

Abstract

Coordination of bacterial stress response mechanisms is critical for long-term survival in harsh environments for successful host infection. The general and specific stress responses of well-studied Gram-negative pathogens like Escherichia coli are controlled by alternative sigma factors, archetypically RpoS.

The deadly hospital pathogen Acinetobacter baumannii is notoriously resistant to environmental stresses, yet it lacks RpoS, and the molecular mechanisms driving this incredible stress tolerance remain poorly defined.

Here, using functional genomics, we identified the transcriptional regulator DksA as a master regulator for broad stress protection and virulence in A. baumannii. Transcriptomics, phenomics and in vivo animal studies revealed that DksA controls ribosomal protein expression, metabolism, mutation rates, desiccation, antibiotic resistance, and host colonization in a niche-specific manner. Phylogenetically, DksA was highly conserved and well distributed across Gammaproteobacteria, with 96.6% containing DksA, spanning 88 families. This study lays the groundwork for understanding DksA as a major regulator of general stress response and virulence in this important pathogen.