Human practices

While talking with different stakeholders during our project, they often asked us ‘What are Human Practices?’ Initially we answered that human practices are interviewing experts on certain topics, and getting their feedback. But as time passed, and many times we had answered this question, we realized that this is not what the true meaning of human practices is. We realized that it is not bombarding academics with questions and awaiting their replies, or asking companies if they would be interested in selling our product, or asking clinicians about how they conduct their clinical practice. It is about listening, about carefully reading between the lines, about looking at the bigger picture. We learned that every person has their unique perspective and story to tell, and it is on us to share it.

Every talk we had was different, with different topics discussed, different opinions, and different questions arising. Each talk was individually assessed, to see how we can implement it into our project, how we can give it a voice, and how we can contribute to bridging the gaps mentioned. Even though every talk was different, most of them agreed on one thing: there is not enough research on PMOS.

*picture of Eleni : Eleni Dubé Zinatelli: ‘I love when you do research on PMOS because honestly, there really isn't enough.’

After many talks, we became increasingly aware of how big the gap can be. It shocked us. Diagnostic delay, lack of proper medication and lack of awareness, just to name a few. They can all be addressed by doing more research on PMOS, because currently, there is no known cause for this syndrome, nor a proper way to treat it. We wanted to bridge that gap, at least a little, because 13% of women suffer from this condition. That is why we dedicated our project to PMOS, to raise awareness about it. There can never be enough awareness.

Stakeholders

Paragraph about different stakeholders.

-insert the circle diagram about different stakeholders-

Integrated HP

Small paragraphs of what decisions we have made to change the project and to implement the feedback from experts. Something similar to a condensed version of all integrations from interviews. Can be ordered in categories possibly.

Academia

Different Academia Consultations

Suman Rice

Reader in Reproductive Physiology at City St George’s, University of London

Suman Rice is a reproductive physiologist specializing in women’s reproductive health, so we contacted her to better understand the hormonal complexity of PMOS and evaluate our androgen targeting approach. She emphasized that PMOS is highly complex and that treatment needs differ depending on symptoms, age, and life stage. Based on her feedback, we decided to define our target patient group and treatment goal more clearly, while paying greater attention to the safety and limitations of androgen reduction, particularly for women who may become pregnant.

Summary of interview:

We contacted Suman Rice because of her expertise in reproductive physiology and women’s reproductive health. We wanted her perspective on whether reducing androgen activity could be a reasonable therapeutic approach for PMOS and what physiological factors we should consider.

We discussed androgen production and regulation in PMOS, the roles of testosterone and aromatase, and limitations of current treatments such as antiandrogen pills and metformin. We also discussed our gut-based approach, suitable target populations, and potential safety concerns.

She emphasized that PMOS involves multiple interconnected hormonal and metabolic pathways, so targeting testosterone alone may not address the whole condition. She also advised us to clearly define which patient group and symptoms we aim to target, and highlighted the importance of considering systemic effects and pregnancy safety when developing an androgen-reducing treatment.

Integration:

Safety: We added pregnancy and fetal safety as an important consideration, since reducing androgen activity could pose risks during pregnancy.

Gut mechanism: We reconsidered how gut localized treatment could affect systemic androgen levels.

Trevor M. Penning

Professor in Pharmacology at University of Pennsylvania

Department of Systems Pharmacology & Translational Therapeutics, University of Pennsylvania Perelman School of Medicine

Trevor M. Penning is an expert in steroid-transforming aldo-keto reductases, including AKR1D1. He was consulted to assess the use of AKR1D1 in our approach, and to discuss potential limitations. He confirmed the enzyme converts testosterone to 5β-DHT quite well, affirming our effector choice. He noted how the AKR1D1’s broad substrate affinity could present an issue, noting that multiple point mutations across the binding pocket would be needed to overcome this challenge, which could pose difficulties within our limited timeline.

Summary of interview:

We consulted Dr. Trevor M. Penning, whose research focuses on steroid transforming aldo-keto reductases, including AKR1D1. He has written many papers on AKR1D1, some of which are considered the backbone of our project. Dr. Penning was consulted to receive feedback regarding the use of AKR1D1 as an effector, and to consult on potential issues, including solubility problems and broad substrate specificity.

Due to other articles having issues with AKR1D1 solubility and most of the protein ending up in inclusion bodies, we discussed a potential solution by adding solubility tags to increase AKR1D1’s solubility. Dr. Penning noted how the formation of inclusion bodies is unavoidable, and the soluble fraction is usually sufficient to do the work, though he emphasized this depends on how much protein is needed. Based on Dr. Benrick’s advice on expressing as much enzyme as possible, we decided to pursue increasing the yield of soluble protein by adding solubility tags.

While discussing issues with substrate specificity, Dr. Penning explained how any substrate with Δ4-3-ketosteroid structure in the steroid ring is a plausible 5β-reduction substrate, which is a primary reason single-steroid specificity is hard to engineer on Δ4-3-ketosteroid reductases. To achieve single steroid specificity, engineering would require multiple point mutations across the substrate pocket. Dr. Penning cautioned us how that would be an extremely difficult task, considering our limited time for the project.

Regarding the question of constitutive or inducible expression for the final product, Dr. Penning pointed us towards bacterial promoters inducible under gut-relevant conditions. Based on his recommendation to pursue a gut-context-inducible promoter, along with Dr. Benrick’s input, we decided to move forward with a hypoxia promoter.

Integration:

Solubility and yield: addition of solubility tags to enzyme constructs was done to maximize the yield of expressed protein

Substrate specificity: decided to pursue multiple point mutations across the binding pocket

Expression system design: choosing a gut-relevant inducible bacterial promotor

Helena Berglund

Head of Teacher Education at Lund University

Joint Faculties of Humanities and Theology, Lund University

Helena Berglund was Head of Teacher Education at Lund University's Joint Faculties of Humanities and Theology, with a research focus on how teacher professionalism can be strengthened through the right conditions in schools and teacher education. Given her expertise, she has insight into how the Swedish school system works and what makes outreach material well-suited for students but also for teachers to implement. We contacted her early on for feedback and support on our high school outreach package, to understand which approaches could realistically create the highest impact. She confirmed our target age (15–18, pre-specialization), pushed us toward hands-on/practical formats over plain lectures, made us consider school segregation, and confirmed language wouldn't be a barrier, given students' English proficiency.

Summary of interview:

Given her research background in teacher professional development, Helena Berglund was contacted to get early feedback on our high school outreach concept and to identify the approaches with the highest impact.

The main topics discussed were: ideal target age, format, the structure of the Swedish school system, the relevance of school segregation, and language requirements. Helena confirmed 15/16 to 18 (pre-specialization and post-specialization = from 9th year onward) as the ideal target age.

She recommended hands-on labs and workshops over lectures alone, as these have a high learning and retention effect on students. She also noted that teachers are generally looking for activities that are easy to fit into an already busy curriculum. She explained the segregation between municipal schools and non-municipal, often profit-driven schools (friskolor) as a reason for unequal access to practical activities, pointing us to Skolinspektionen for insight into school quality data and suggesting we could prioritize socioeconomically weaker areas as high-impact targets. She confirmed English-language material is fine given students' strong proficiency. She also highlighted linking science to its societal relevance as the most engaging angle for student interest.

Integration:

Format: She strengthened our decision to develop a lecture + workshop package rather than a lecture alone. We also kept this in mind across almost all educational talks/lectures, integrating at least some interactive component into each one. As one example, following her advice to prioritize practical activities, we replaced a planned lecture with a lab exercise for the Rays for Excellence event (LINK), even though a lecture had originally been suggested.

Material accessibility: Based on her point about school segregation and unequal access to resources, we kept material low-cost and accessible. For example, using pasta rings to illustrate plasmid parts (promoter, etc.) in the BioCamp lecture + workshop (LINK).

Language: Her reassurance that English is widely understood by Swedish students let us proceed with the high school package in English, which was necessary at the time since we had no Swedish-speaking team member yet.

Content framing: Her advice to link science to its societal relevance reinforced the approach we started with, using iGEM to show the range of experts involved in synthetic biology (ethics, policymakers, researchers, etc.).

Anna Benrick

Professor in endocrinology at University of Gothenburg

Department of Physiology, University of Gothenburg

Anna Benrick’s research focuses on hyperandrogenism and its link on reproductive diseases in females. She was contacted to help us with understanding more about PMOS, hyperandrogenism and the feasibility of our approach. Dr. Benrick confirmed 5β-reductase-targeted androgen reduction is a valid therapeutic approach, and advised us against androgen inducible expression strategy, favoring stable overexpression instead.

Summary of interview:

We consulted Dr. Anna Benrick, whose research focuses on the molecular link between androgen excess and the development of reproductive disorders, obesity, and type-2 diabetes in women. The interview was centered on the feasibility of the AnTigone project, whether locally reducing testosterone in the gut could meaningfully affect systemic testosterone levels and reduce hyperandrogenism symptoms. Despite promising preclinical findings, Dr. Benrick emphasized that the gut microbiota-PMOS field is still young, and it remains unclear whether microbiota changes drive PMOS or result from it.

She pointed out a recent paper on artemisin, an antimalarial compound shown to lower testosterone production by inhibiting a testosterone synthesis enzyme, suggesting an alternative approach to reducing systemic testosterone levels. A recurring topic in the interview was the uncertainty about gut-to-serum steroid circulation: how much testosterone actually passes through the gut relative to what is produced by ovaries and adrenal glands. This uncertainty was raised to answer one of our main questions, whether our approach could achieve a biologically relevant effect. We had discussed the possibility of incorporating a testosterone-sensing mechanism to induce enzyme expression and deactivate it if reduction became too high. She advised not to pursue this, but to prioritize stable, strong enzyme expression, since insufficient effect was deemed more likely to happen than excessive androgen lowering. Furthermore, she did not anticipate a compensatory increase in androgen production in response to our treatment.

The conversation also touched on validation strategy, including in vitro options such as testosterone-producing theca cells, and candidate biomarkers such as testosterone, androstenedione and hydroxyandrogens, alongside clinical symptoms such as menstruation frequency or body hair. Dr. Benrick raised the possibility that a gut-localized, low-risk approach could be relevant for PMOS treatment during pregnancy, a period in which most hormonal therapies are avoided due to their possibility of affecting fetal development.

Integration:

Expression system design - adopting a hypoxia-responsive promoter as a fixed gut on/off switch

Project framing - framing the project as an early-stage hypothesis rather than a validated therapeutic mechanism

Clinicians

Different Clinician Consultations

Industry

Different Industry Consultations

Clinician survey

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Self scoring

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