As an interdisciplinary team of classically trained biochemists, cell biologists and engineers, we look for approaches that combine answering fundamental questions with cutting-edge practical applications. Nowhere is this combined potential higher than in hydrogel research. Biological “living” hydrogels are self-assembled systems made from highly hydrated biopolymers. They coat all the wet surfaces in the human body, providing a selective barrier that allows nutrients and information in while keeping pathogens out. One example of a hydrogel-based filter is the mucus barrier, the slimy gel that controls the exchange of particulates and microbes between the body and the environment. From an engineer’s perspective, we believe hydrogels can solve some vexing problems, such as passivating materials implanted into the human body, preventing bacterial biofilm formation on materials exposed to living organisms, and selectively filtering biomolecules. From a biochemist’s perspective, our goal is to elucidate the fundamental biophysical principles that allow biogels to act as selective barriers and filters, and the cellular mechanisms involved in building and regulating them.
Mucus is an important and understudied biological hydrogel that lines an enormous surface area in our body (2000 square feet in the intestine alone). Mucus is an ideal system in which to elucidate general principles of bio-filtration by hydrogels. It is also of great practical importance to many engineering applications relating to the human body. Any device or drug in the gut, the reproductive tract or the eye is immersed in mucus, and its performance will be largely defined by this interaction. Moreover, mucus is home to trillions of microbes that form our microbiome and regulates their interactions with the host. Yet, mucus is little studied by biologists or engineers.
The gel-forming building blocks of mucus are long and threadlike polymers called mucins. Mucins look like tiny bottlebrushes and are grafted with a myriad of unique sugar structures, creating a wealth of biochemical information. Although the exceptional molecular diversity and complexity of mucin-associated glycans has been recognized for some time, their regulatory potential has barely been tapped because their individual bioactivities have been difficult to analyze.
Our lab has developed a suite of methods to analyze native mucus, hydrogels built from purified mucins, and isolated mucin glycans. The focus of our lab is on basic mechanisms by which mucus interacts with pathogens and shapes microbial communities, the mechanisms by which mucus barriers exclude, or allow passage of different molecules and pathogens, as well as the link between dysfunctional mucus barriers and disease. We hope to provide the foundation for a theoretical framework that captures general principles governing selectivity in mucus, and likely other biological hydrogels such as the extracellular matrix, and bacterial biofilms. The lab’s work may also be the basis for the reconstitution of synthetic polymers and gels that mimic, or are superior to, their natural counterparts.
Mucus and Microbes
An integral arm of our research is to better understand how the human body prevents infections by harmful microbes on mucosal surfaces. Mucus is one major ecological niche for the human microbiota – it accommodates trillions of microbes where densities approach 1011–1012 cells/mL, a record for any microbial ecosystem documented so far. Many of these microbes are important for our health because they help us build vitamins and digest food, and some can protect us from other damaging microbes. But in this crowd are also numerous potentially harmful pathogens that can cause infections. It is a wonder we are alive! It appears that over millions of years, the mucus has evolved the ability to control problematic pathogens, preventing them from causing damage, but studies that directly test this aspect of mucus are missing.
Over the past years our laboratory has developed laboratory models for mucosal surfaces to study the effects of mucus and mucins, the major gel-forming constituents of the mucus barrier, on the behavior of prevalent opportunistic pathogens, including Pseudomonas aeruginosa, Candida albicans, and Streptococcus mutans. We have discovered that the mucus has a profound influence on the behavior of microbes and specifically, their ability to swim, to settle, and to communicate with each other, and that it can suppress a range of virulence traits including surface attachment, biofilm formation, and horizontal gene transfer. Our work furthermore shows that the mucus creates an environment, which stabilizes the viability of microbes while preventing them from doing harm, analogous to the process of microbe domestication. This is important – mucus does not control pathogens by killing them, but instead appears to render them docile. Our team has created this TEDEd video and other productions (see media page) to explain the fascinating functions of mucus.
Research on mucus makes a fundamental contribution to our understanding of host-microbe interactions as it is generating a paradigm shift from the textbook view of mucus as a simple catchall filter for particles, toward the understanding that is a sophisticated bioactive material with powerful abilities to manipulate microbial behavior. In addition, by introducing well-defined in vitro laboratory systems, as developed in our and other laboratories, we can bring a big push to a field that lacks tools for the mechanistic analysis of mucus-microbe interactions.
Understanding the basic science of biological problems underlies all our endeavors, but we also strive to find cutting-edge practical applications for our work. One application with particularly high translational potential lies in using mucus as a non-invasive diagnostic for epithelial health. The rationale is that the physicochemical properties of mucus barriers are intricately related to health and disease, and the pathological onset of mucus barrier dysfunction can lead to a number of devastating pulmonary, gastrointestinal and urogenital conditions.
Over the past years our laboratory has begun to unravel the strong diagnostic potential of mucus by interrogating the physicochemical properties of cervical mucus in the context of preterm birth. Preterm birth affects about 13 million babies worldwide each year, resulting in major pediatric morbidity and mortality, as well as significant health care costs. In a significant number of cases, preterm birth correlates with increased ascending bacterial infections from the female vaginal tract, suggesting that the cervical barrier to infection is weakened. Our study reveals that cervical mucus from women at high risk for preterm birth is more translucent, extensible, and permeable, than cervical mucus from women in a healthy pregnancy, suggesting that a weakening of the mucus barrier may be one main contributor to intrauterine infection. This discovery not only allows for patient stratification, but it also provides valuable mechanistic insights into the causes of preterm labor.
Looking forward, we are applying our tools to diagnose diseases on other mucosal surfaces, including the oral cavity, the intestinal linings, and the lungs, with the goal to establish a structure-function relationship between mucus’ physicochemical properties and epithelial health. We anticipate that our work will have important implications for understanding disease progression on mucosal epithelia, may enable the more rapid development of diagnostic tools that utilize mucus as a source of biophysical indicators for health and disease, and possibly direct the design of intervention strategies.
Little is known about the detailed molecular properties that distinguish particles that permeate, or are rejected by, a defined mucus barrier. What is the relationship between a particle’s size, geometry and biochemistry, and its mobility across a mucus barrier? Can we build a physical model of hydrogels that predicts how their structure governs filtration properties? How do certain pathogens such as viruses or bacteria penetrate mucus barriers? To address these central questions we have developed new technology and concepts to quantify the uptake, transport and spatial distribution of particulates and cells in mucus.
Our research shows that mucus gels provide an effective and selective barrier toward particles as diverse as protons, antibiotic molecules, peptides, and even toward a broad range of viruses. Our work suggests that the mucus hydrogel does not operate predominantly on the basis of size exclusion, a mechanism that is often suggested in text books, but rather as a sophisticated interaction filter in which binding of bypassing particles by the hydrogel polymer components is used to govern transport through the hydrogel. Specifically, we can conclude that commonly considered parameters such as net charge or lipophilicity are insufficient predictors for mucus permeability, and instead, the detailed spatial distribution of charged and hydrophobic groups can strongly modulate diffusivity. Over the next years we shall determine the rules for how the detailed biochemistry and structure of particles determine their passage through hydrogels. Mucus in the gastrointestinal, respiratory, and reproductive tracts are obvious sites for delivering therapeutics. Understanding the basic principles that govern its selectivity may enable the design of small molecules and vectors for drug delivery, and new approaches for preventing prevalent infectious diseases (e.g., HIV, Zika, COVID-19), or unwanted pregnancy. Our long-term aim is to provide the foundation for a theoretical framework that captures general principles governing selectivity in mucus, and likely also in other biological hydrogels such as the extracellular matrix, and bacterial biofilms.