{% set sections = [ {"icon": "fa-solid fa-compass", "title": "Our Story"}, {"icon": "fa-solid fa-suitcase-medical", "title": "Endometriosis"}, {"icon": "fa-solid fa-heart", "title": "Human Practices"}, {"icon": "fa-solid fa-computer", "title": "Dry Lab"}, {"icon": "fa-solid fa-bacteria", "title": "Wet Lab"}, {"icon": "fa-solid fa-book", "title": "References"}, ] %} {% extends "window-layout.html" %} {% block title %}Project Description{% endblock %} {% block lead %}Introduction to project LactoBack.{% endblock %} {% block window_content %}

Recent research suggests that a healthy vaginal microbiome is defined by the dominance of a single bacterial species, the probiotic Lactobacillus crispatus being one of the most frequent dominant species. In an unhealthy microbiome, this dominance is weakened.

Also known as vaginal dysbiosis, this condition is often associated with endometriosis.


Our therapy aims to restore the healthy state of the vaginal microbiome by promoting the growth of Lactobacillus crispatus with a self-sustaining bacterial system. Our helper bacterium would return the naturally dominant bacterium its competitive edge by conditionally secreting lactic acid, which lowers the vaginal pH and hampers the growth of opportunistic bacteria.

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Even though endometriosis affects up to 10% of women, causing severe pain and resulting in repeated operations, the condition has no cure and no satisfactory treatments; it is little-known despite the high proportion of people affected, and in fact, we don't know what causes it.

In the UZurich iGEM team, we want to address this from an often-overlooked perspective.

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{{ sections[0].title }}: What Inspired Us


In our team, almost everyone knows someone suffering from endometriosis. This project, for us, tackles an issue as personal as it is global.
We believe women deserve a timely diagnosis and appropriate treatment. We believe women shouldn't find it normal to be in pain.

{{ sections[1].title }}: The Problem


In endometriosis, some of the cells normally found inside the uterus are also found outside the uterus, where they undergo a monthly cycle, as if they were in the uterus, and shed. This leads to progressively worsening internal inflammation, scarring, cysts and extreme pain. Currently, the main therapies consist of hormonal therapies and operations, both of which are very invasive. One of the difficulties to develop a good therapy is that we don't know what causes the disease.
At the moment, a lot of research is going into applying cancer therapies to endometriosis, since, like cancer cells, endometriosis cells are cells that are found in the wrong place (keyword ectopic tissue). However, one avenue of treatment that has received little attention so far is the role of the microbiome in endometriosis. There are multiple indications that a woman's microbiome - vaginal and gut - are important in the development of endometriosis.
In fact, an imbalanced “unhealthy” vaginal microbiome (with what is known as vaginal dysbiosis) is often associated with endometriosis. While recent research suggests that a “healthy” vaginal microbiome is dominated by a single bacterial species, such as the probiotic bacterium Lactobacillus crispatus, this dominance is decreased in the “unhealthy” microbiome.
Thus, our therapy aims to restore the healthy state of the vaginal microbiome by promoting the growth of Lactobacillus crispatus, one of the most frequent dominant species, with a self-sustaining bacterial system.

{{ sections[2].title }}: Making the Invisible Visible


Awareness of gender inequalities in medicine are on the rise, and now is the moment to educate the public and take initiative to find an innovative treatment. As we've worked to publicise our project and speak with Swiss patients and doctors, we've seen repeatedly how important it is to those with endometriosis to finally be heard.
Our vision of human practices is based on three pillars: awareness, synthetic biology & education, and integrated human practices, with the aim to improve the lives of women suffering from endometriosis.

Awareness

To educate the public about endometriosis and gender inequalities in medicine, we

    ...have surveyed patients and experts, receiving over 200 answers.
    ...are interviewing patients and doctors.
    ...are organising our Lunch Lecture Series with experts from various fields including medicine and psychology to discuss medicinal gender inequality.
    ...use social media channels to educate people about the disease and the patients' experience an communicate with medical, scientific and activist organizations.
    ...are publishing articles, e.g. in youth magazines, to educate the public.

Education in Synthetic Biology

At the same time, we want to bring synthetic biology closer to the students and the public, so we are:

    …organising a workshop series in high schools to introduce the pupils to synthetic biology and its potential to combat global problems.
    …organising a lab workshop series with patients to show them how synthetic biology can be applied to find novel treatments for on disease.

Integrated Human Practices

Finally, we are supporting the wet lab by speaking with experts and digging into current research, giving our team members feedback about the needs of patients and doctors. The goal is to design a model that suits the patients' needs as well as possible, which can only happen in direct contact with society.

{{ sections[3].title }}: Setting Targets, Going Beyond


The dry lab team helps streamline the project by reducing the wet lab's experimental load and by simulating and predicting how our system would perform in real patients.

Working Hand-in-Hand With the Wet Lab

Looking a few steps ahead

We help with the selection of gene targets for flux redirection via flux balance analysis (FBA) - a genome-scale model of reactions and metabolites present in the cell - to determine the most useful combinations of genes to conditionally knock down in the wet lab. The results of our simulations are then used to engineer dCas9 sgRNA-based parts. To verify that our engineered parts are robust and functional, we test them using ODE modelling with the Simbiology Toolkit of MATLAB.
Throughout the project, we also verify our model predictions and effects in vitro with metabolic flux analysis (MFA) using mass spectrometry-assisted metabolomics.

There are many things our wet lab team would find difficult to accomplish in vitro. However, with the power of computational modelling, we can extend what the wet lab has done. Here, we investigate how our engineered bacteria would behave in our target environment and how they would react to various invaders of the microbiome.
We do this by modelling competitive bacterial growth over a period of time, looking at how our bacterial system would switch between different states, to test the system's robustness and resilience and give feedback on how it could be improved.

{{ sections[4].title }}: Targeting an Imbalanced Vaginal Microbiome


We hope to reestablish a healthy microbiome by engineering a self-sustaining helper bacterium population that grows well in dysbiotic - i.e. unhealthy - vaginal conditions. Our helper bacterium would give Lactobacillus crispatus, the dominant bacterium in many women, a competitive edge by conditionally secreting lactic acid, which lowers the vaginal pH and hampers the growth of opportunistic bacteria.



The LactoBack System:

LactoBack E. coli system schematic
nothing here yet...

Figure 1: Left: Overview of the engineered E. Coli bacterium: Rhl-QS molecules are depicted in violet. Right: Depiction of a hypothetical timeline from: eubiotic conditions (1), disruption that leads to dysbiosis (2), growth of engineered E. coli bacterium that leads to secretion of lactate and thus lowering of pH (3) and reestablishment of eubiotic conditions (4).

At the start of dysbiosis Lactobacillus crispatus greatly reduces in number. Simultaneously there is a strong increase in pH and opportunistic bacteria, including the engineered helper bacterium take-over. The level of C4-HSL rises with an increasing number of helper bacteria. Once a threshold level is reached, metabolic flux is redirected to produce great quantities of lactic acid. Opportunistic bacteria can no longer compete with Lactobacillus crispatus and a healthy environment is reestablished.


We plan to introduce a quorum sensing system into our helper bacterium that lets it detect its own population density (Fig. 1) so that lactic acid production is only triggered at high densities. This precise live response to the patient's current state, combining monitoring and treatment, is only possible with precise engineering using synthetic biology.
In the future we aim to engineer the same system into a variety of vaginal bacteria, which would allow for personalised medicine, to reflect the different makeup of the vaginal microbiome in women of different ethnicities.

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    [1] Alakomi, H.-L. et al. Lactic Acid Permeabilizes Gram-Negative Bacteria by Disrupting the Outer Membrane. Appl Environ Microbiol 66, 2001-2005 (2000).
    [2] Amabebe, E. & Anumba, D. O. C. The Vaginal Microenvironment: The Physiologic Role of Lactobacilli. Frontiers in Medicine 5, (2018).
    [3] Boris, S. & Barbés, C. Role played by lactobacilli in controlling the population of vaginal pathogens. Microbes and Infection 2, 543-546 (2000).
    [4] Happel, A.-U. et al. Exploring potential of vaginal Lactobacillus isolates from South African women for enhancing treatment for bacterial vaginosis. PLoS Pathog 16, e1008559 (2020).
    [5] Kalia, N., Singh, J. & Kaur, M. Microbiota in vaginal health and pathogenesis of recurrent vulvovaginal infections: a critical review. Annals of Clinical Microbiology and Antimicrobials 19, 5 (2020).
    [6] O'Hanlon, D. E., Moench, T. R. & Cone, R. A. Vaginal pH and Microbicidal Lactic Acid When Lactobacilli Dominate the Microbiota. PLOS ONE 8, e80074 (2013).
    [7] Ravel, J. et al. Vaginal microbiome of reproductive-age women. Proceedings of the National Academy of Sciences 108, 4680-4687 (2011).
    [8] Richeux, V. Endometriosis Linked With Genital Microbiome Changes. Medscape https://www.medscape.com/viewarticle/983197 (2022).
    [9] Ser, H.-L., Au Yong, S.-J., Shafiee, M. N., Mokhtar, N. M. & Ali, R. A. R. Current Updates on the Role of Microbiome in Endometriosis: A Narrative Review. Microorganisms 11, 360 (2023).
    [10] Uzuner, C., Mak, J., El-Assaad, F. & Condous, G. The bidirectional relationship between endometriosis and microbiome. Frontiers in Endocrinology 14, (2023).
    [11] Wang, S. et al. Antimicrobial Compounds Produced by Vaginal Lactobacillus crispatus Are Able to Strongly Inhibit Candida albicans Growth, Hyphal Formation and Regulate Virulence-related Gene Expressions. Frontiers in Microbiology 8, (2017).
    [12] WHO. Endometriosis. https://www.who.int/news-room/fact-sheets/detail/endometriosis.
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