Tuberculosis thrives in the lungs, but when the bacteria causing the disease are expelled into the air, they face a much harsher environment with drastic changes in pH and chemistry. Understanding how these bacteria survive this airborne journey is essential for their persistence, yet little is known about the mechanisms that protect them as they move from one host to another.
Now, MIT researchers and their collaborators have identified a family of genes that are crucial for the bacterium’s survival specifically when exposed to the air, likely offering protection during its transmission.
Previously, many of these genes were thought to be nonessential, as they didn’t appear to affect the bacteria’s role in causing disease when introduced into a host. This new study, however, suggests that these genes are vital for transmission rather than proliferation.
“There is a blind spot in our understanding of airborne transmission, especially regarding how a pathogen survives sudden environmental changes as it circulates in the air,” says Lydia Bourouiba, head of the Fluid Dynamics of Disease Transmission Laboratory, associate professor of civil and environmental engineering, mechanical engineering, and core faculty member at MIT’s Institute for Medical Engineering and Science. “Now, through these genes, we have an insight into the tools tuberculosis uses to protect itself.”
The team’s findings, published this week in Proceedings of the National Academy of Sciences, could lead to new tuberculosis therapies that target both infection and transmission prevention.
“If a drug targeted the products of these genes, it could effectively treat an individual and, even before that person is cured, prevent the infection from spreading,” says Carl Nathan, chair of the Department of Microbiology and Immunology and the R.A. Rees Pritchett Professor of Microbiology at Weill Cornell Medicine.
Nathan and Bourouiba co-senior authored the study, which includes MIT collaborators and Bourouiba’s mentees from the Fluids and Health Network: co-lead author postdoc Xiaoyi Hu, postdoc Eric Shen, and students Robin Jahn and Luc Geurts. The research also involved collaborators from Weill Cornell Medicine, the University of California at San Diego, Rockefeller University, Hackensack Meridian Health, and the University of Washington.
Pathogen’s Perspective
Tuberculosis is a respiratory disease caused by Mycobacterium tuberculosis, a bacterium that predominantly affects the lungs and spreads through droplets released by an infected person, typically when they cough or sneeze. Tuberculosis remains the leading cause of death from infection, except during global viral pandemics.
“In the last century, we’ve seen the 1918 influenza pandemic, the 1981 HIV/AIDS epidemic, and the 2019 SARS-CoV-2 pandemic,” notes Nathan. “Each virus has caused significant loss of life, but after they subsided, we were left with the ‘permanent pandemic’ of tuberculosis.”
Much of the research on tuberculosis focuses on its pathophysiology—how the bacteria infect a host—along with diagnostic and treatment methods. For their new study, Nathan and Bourouiba turned their attention to tuberculosis transmission, specifically exploring how the bacteria defend themselves during airborne transmission.
“This is one of the first efforts to study tuberculosis from an airborne perspective, investigating how the organism survives harsh changes in the environment during transmission,” says Bourouiba.
Critical Defense
At MIT, Bourouiba studies fluid dynamics and how droplet behaviors can spread particles and pathogens. She partnered with Nathan, who investigates tuberculosis and the genes that the bacteria rely on throughout their life cycle.
To explore how tuberculosis survives in the air, the team aimed to replicate the conditions the bacterium encounters during transmission. They first worked to develop a fluid with similar viscosity and droplet sizes to those expelled by a person coughing or sneezing. Bourouiba points out that previous research on tuberculosis relied on liquid solutions that are used to grow the bacteria. However, these liquids differ significantly from the fluids tuberculosis patients expel.
Furthermore, the fluid typically sampled from tuberculosis patients, like sputum for diagnostic tests, is thick and sticky, which makes it inefficient at spreading and forming inhalable droplets. “It’s too gooey to break into inhalable droplets,” Bourouiba explains.
Through her research on fluid and droplet physics, the team determined a more accurate viscosity and droplet size distribution for tuberculosis-laden microdroplets in the air. They also analyzed the composition of droplets by studying infected lung tissue samples. They then created a fluid that mimicked the viscosity, surface tension, and droplet size that would be released into the air when a person exhales.
Next, the team deposited different fluid mixtures onto plates as tiny droplets, measuring how they evaporated and what structures they left behind. They discovered that the new fluid shielded the bacteria at the center of the droplet, unlike traditional fluids where bacteria were more exposed to the air. The realistic fluid also retained more water.
The team then infused the droplets with bacteria carrying genes with various knockdowns to see how the absence of specific genes affected bacterial survival during evaporation.
They evaluated over 4,000 tuberculosis genes and identified a family of genes that became crucial in airborne conditions. Many of these genes are involved in repairing damage to oxidized proteins, such as those exposed to air, while others are responsible for breaking down irreparably damaged proteins.
“What we found is a lengthy list of candidate genes, some more prominently involved than others, that could play a critical role in helping tuberculosis survive during transmission,” Nathan says.
While the experiments cannot fully replicate the bacteria’s biophysical transmission (as droplets fly through the air and evaporate), the team mimicked these conditions by placing plates in a dry chamber to accelerate droplet evaporation, similar to what happens in flight.
Going forward, the researchers are developing platforms to study droplets in flight under various conditions. They plan to further investigate the role of the newly identified genes in more realistic experiments, potentially weakening tuberculosis’s airborne defenses.
“The idea of waiting to diagnose and treat someone with tuberculosis is an inefficient way to stop the pandemic,” Nathan says. “Most individuals who exhale tuberculosis haven’t been diagnosed yet, so we need to interrupt its transmission. Understanding the process is key, and now we have some ideas.”