Why He Was Contacted:

As a recognized expert in urology, we approached Dr. Golomb for in-depth insights into the current treatments for rUTIs, the challenges patients face when afflicted by the infection, and his firsthand experiences in managing it. Our goal was to garner a holistic understanding of the condition, starting with a medical perspective, before eventually integrating the patient's viewpoint to paint a comprehensive picture.

What We Learned:

In our discussion with Dr. Golomb, we gained insights into the primary treatment approaches for UTIs. We learned that the predominant treatment is the use of antibiotics, as advised by the urologists. In certain instances, other treatments such as Hyprex antibacterial medications or hormonal treatments using estrogen may be considered. For recurrent UTIs, the treatment often involves continuous, low-dose antibiotic administration over several months. Additionally, we delved into the profound impact of rUTIs on patients' lives, encompassing symptoms such as frequent urination during the day, discomfort, and social challenges. Dr. Golomb also highlighted the potential of Lactobacillus-based probiotics, noting many UTI patients find relief with probiotic supplementation.

What We Did:

The pivotal realization stemming from our discussion was the pervasive dependence on antibiotics, a trend that urged us to explore innovative alternatives for UTI treatment. Our objective became clear: we aimed to discover a UTI treatment that could bypass the conventional use of antibiotics.

Why She Was Contacted:

Given her extensive research into the female microbiome and women's health, Professor Achinoam Lev-Sagi emerged as a pivotal expert for our consultations. We sought to pinpoint the optimal bacterium that could be modified and harnessed as a solution for recurrent Urinary Tract Infections (rUTIs).

What We Learned:

Through our discussions, we aimed to identify a bacterium that's non-pathogenic and naturally present in the vaginal microbiome. Prof. Lev-Sagi confirmed that Lactobacillus Crispatus (L. Crispatus) is a fitting bacterium for our objectives. She underscored the imperative nature of modifying the vaginal and urinary bacterial compositions to effectively counteract rUTIs. This is grounded in the intricate relationship between the urinary and vaginal microbiomes in women: disruptions in one can influence the other, sometimes leading to UTIs. Prof. Lev-Sagi emphasized the importance of understanding and addressing this dynamic for holistic women's healthcare. She also touched upon the essentiality of an uncomplicated administration method for any developed product. Oral medicine, she noted, is the simplest and most preferred method among patients.

What We Did:

As a result of this discussion, we made the decision to proceed with engineering L. Crispatus. We chose it over other candidates that had previously been under consideration, thus finalizing our selection based on the expert advice provided.

Why He Was Contacted:

Ethical considerations were a top priority throughout our entire project, ensuring comprehensive integration from the initial planning stages to the survey phase. To navigate the intricate ethical landscape of our research, we consulted with Dr. Maya Peled Raz, an expert in bioethics whose guidance was invaluable in shaping our ethical framework during the survey phase.

What We Learned:

Dr. Maya Peled Raz emphasized the ethical considerations we needed to address regarding our survey, focusing on ensuring the safety and confidentiality of the gathered data. We meticulously assessed potential harms associated with our survey, taking careful measures to identify and safeguard any groups that might be endangered or upset by it or our research.

What We Did:

As a result, we adjusted the survey to make sure it follows the ethical guidelines suggested by Dr. Maya Peled Raz. We informed the participants that by answering the survey, they indicated their agreement to participate. Additionally, we took measures to ensure the survey was anonymous, and the collected data was securely stored.

Why He Was Contacted:

We sought Dr. Michael Cohen's expertise due to his extensive experience in urology and urological oncology. Given his leadership in the Urology Department at HaEmek Medical Center, Afula, we believed he could provide valuable insights on UTIs, existing treatments, their mechanisms of action, and the most effective methods of product delivery for our proposed solutions.

What We Learned:

Dr. Cohen offered a thorough insight into the intricacies of UTIs, highlighting specific risk groups and elaborating on current treatment methodologies. He underscored the relationship between these risk groups and the emergence of antibiotic resistance. When addressing our proposed solution, he advocated for local delivery techniques, stressing the benefits of suppositories or vaginal inserts for their targeted application and minimal systemic implications. This perspective stood in contrast to Prof. Lev-Sagi's proposal of oral administration.

What We Did:

Based on the insights from both Dr. Cohen and Prof. Lev-Sagi, we recognized the need for more definitive directions regarding our delivery method. To gain a clearer understanding, we decided to consult with an expert in the field, Prof. Boaz Mizrahi.

Why He Was Contacted:

After receiving conflicting perspectives from Dr. Michael Cohen and Prof. Achinoam Lev-Sagi regarding drug delivery methods for our product, it became evident that further expert consultation was essential. Prof. Boaz Mizrahi, with his distinguished expertise in biotechnology, pharmaceutics, and specifically drug delivery, was approached to provide clarity and guidance on the most suitable delivery method for our product. His profound understanding of the field was sought to navigate the complexities and ensure the efficacy of our solution.

What We Learned:

During the discussion with Prof. Mizrahi there was a crucial emphasis on establishing a clear connection between the introduction of bacteria into the digestive system and subsequent improvements in the urinary system. Various delivery methods, ranging from traditional capsules to innovative suppositories, were explored, each with its set of advantages and challenges. Prof. Mizrahi highlighted the importance of comprehensive microbial sampling both before and after drug administration, providing valuable insights into the research methodology for our drug delivery method. Additionally, the potential of utilizing a spray containing the specific bacteria was discussed, presenting a novel approach to bacterial delivery. A focal point of the discussion was the functionality of Lactobacillus Crispatus, shedding light on its potential applications in the context of urinary health. Furthermore, the feasibility of focusing on genetically modified organisms (GMOs) was discussed, accompanied by suggestions on how to address regulatory concerns and safety issues effectively.

What We Did:

Ultimately, after reviewing the myriad of options presented by Prof. Mizrahi and considering various research papers, we recognized the importance of thorough clinical research to determine the feasibility and efficacy of each potential delivery method. As a result, we've decided to earmark the finalization of the delivery method as a future endeavor.

Why She Was Contacted:

In order to understand how to overcome the pathogenicity of E. coli and design our UTI cure, we needed to comprehend the dynamics of E. coli. Furthermore, we wanted to understand the effects a genetically modified lactobacillus could have in the body while fighting E. Coli. For this purpose, we consulted with Dr. Anat Shteren. Her expertise in infectious diseases and internal medicine made her the appropriate expert for our research needs.

What We Learned:

During our meeting, we gained insights into the pathogenicity of E. coli which proved instrumental in guiding the design of our L. Crispatus to effectively counteract E. coli. Furthermore, Dr. Shteren emphasized the issue of antibiotic resistance, underscoring the paramount importance of finding alternative solutions. Finally, the potential pathogenicity of our proposed solution was discussed, and Dr. Shteren concluded that the only way of knowing for sure wether our solution would cause an inlamatory response would be in the clinical trials stage of our research, beyond the scope of iGEM. She explained what tests we’d need to do and who the test subjects should be.

What We Did:

Following our discussion with Dr. Anat Shteren, we took a deep dive into researching the effects of altering the adhesive properties of L. Crispatus to express E. coli adhesive proteins. Recognizing the potential implications on pathogenicity, we resolved to consult more experts in the field for a comprehensive understanding. Subsequently, we reached out to Prof. Yoram Reiter, a distinguished immunologist and a front-runner in immunology and cancer research, as well as Prof. Sima Yaron, a molecular biologist with a specialization in foodborne pathogens.

Why He Was Contacted:

At the point of reaching out to Prof. Yoram Reiter, our proposed strategy involved expressing FimH, Uropathogenic E. Coli (UPEC)'s primary adhesive protein, on our Lactobacillus' membrane. This approach aimed to compete with E. Coli, thereby mitigating and preempting infections. Given Prof. Reiter's deep expertise in immunology, we sought his perspective to grasp the potential ramifications of FimH expression, particularly concerning pathogenicity and immunogenic outcomes.

What We Learned:

From our conversation with Prof. Reiter, we grasped the implications of expressing FimH on the bacteria's membrane. He underscored the potential pathogenic risks this could introduce, casting doubts about its therapeutic safety. Moreover, he indicated that secretions, which could be a potential way to avoid pathogenicity, would likely be impactful only at significantly high concentrations, highlighting potential hurdles in its practical deployment.

What We Did:

Following Prof. Reiter's insights, the FimH membranal expression idea was abandoned, and experiments were designed to determine the optimal concentration thresholds for secretion. Simultaneously, we delved into the competitive innate mechanisms of adherence proteins on Lactobacillus crispatus through in-depth literature reviews. Additionally, we sought additional strategies to combat E. Coli, expanding our exploration beyond just competition using our Lactobacillus chassis.

Why She Was Contacted:

Prof. Sima Yaron was contacted due to her specialization in understanding the virulence and resistance mechanisms of foodborne and waterborne human pathogens. We sought her insights to unravel the complexities of the relationship between elevated bacterial adhesion and the body's immune response. Additionally, her knowledge was pivotal in addressing the critical issue of antibiotic overuse across various sectors, especially in the context of its impact on both healthy individuals and the immunocompromised.

What We Learned:

Prof. Yaron mentioned the intricate dynamics between elevated bacterial adhesion and the body's immune response. She provided insights, highlighting that while heightened adhesion can bolster immunity, it does not necessarily signify direct pathogenicity. This nuanced perspective added depth to our comprehension of host-pathogen interactions. Furthermore, our discussion illuminated the alarming consequences of antibiotic overuse, emphasizing the heightened risks it poses to both the general population and those with compromised immune systems, reaffirming the need for a non antibiotic-based solution to rUTIs.

What We Did:

Guided by Prof. Yaron's insights, we delved deeper into literature pertaining to the adhesion mechanisms of FimH. Our extensive research solidified our conviction that secretion, rather than direct expression of the protein, would be the most beneficial approach. Acknowledging the crucial need for patient perspective and its role in shaping our endeavor, we transitioned to the subsequent phase of our human practices efforts. This involved launching a comprehensive survey and actively consulting with our primary stakeholders — the patients — to gather firsthand insights and feedback on our proposed solutions.

Survey

Through a comprehensive survey, we investigated the prevalence of urinary tract infections (UTIs) in women, exploring the profound impact of these infections on the daily lives of affected individuals. Additionally, we analyzed a variety of treatments, evaluating their effectiveness and accompanying side effects. Moreover, the survey assessed public thoughts and awareness concerning antibiotic resistance and genetically modified organisms (GMOs). The primary objective was to acquire a nuanced understanding of the intricate challenges associated with UTIs.

UTI's Prevalence and Effect

The survey, involving 255 participants among which 213 are women, presented disconcerting findings which reinforced our commitment to addressing the challenge of UTIs. The data revealed that 33% of surveyed women had encountered at least one UTI within the preceding six months. Notably, 24% of these women reported a need to urinate 9-10 times daily, and an alarming 44% had to visit the restroom over 10 times a day. Furthermore, the survey illustrated that UTIs had a significant impact on 57% of women concerning sexual intercourse, and 67% reported interference with their ability to study. These results underscore the adverse effects of UTIs on women's overall well-being and highlight the debilitating consequences of UTIs in the context of a woman's daily life.

Today's Treatments

The survey highlighted the disconcerting reliance on antibiotic treatments, with 55% of UTI sufferers resorting to this method, thereby contributing to concerns surrounding antibiotic resistance within the scientific community. It is important to note that UTIs recur even after consuming antibiotics, demonstrating the limitations of antibiotics as a definitive solution for UTIs. Additionally, 35% of participants experienced side effects such as fungal infections due to standard treatments, further emphasizing the urgent need for efficient and safe therapeutic options. Moreover, the survey revealed that 77% of UTI sufferers who consumed probiotic products did so after taking antibiotics, indicating an attempt to recover from the effects of antibiotic treatment. These findings highlight that solely relying on probiotics cannot provide a comprehensive solution to the problem. Despite doctors' testimonies indicating that UTI patients do use probiotics, this approach alone proves insufficient in addressing the issue and gives our research project an advantage over probiotics.

Treatment's Efficiency

The assessment of UTI treatments efficacy raised significant concerns. Current treatment methods received a high score of 3.92 out of 5 for short-term efficiency. As previously mentioned, 55% of UTI sufferers resort to antibiotics. However, for long-term treatment efficacy, respondents rated it at 2.82 out of 5. These results suggest a tendency toward increased antibiotic usage for immediate relief. Consequently, this trend could further diminish the long-term treatment efficiency and pose a substantial threat, hindering our progress in addressing broader scientific challenges, particularly antibiotic resistance.

Public Thoughts and Awareness

Public awareness to the global issue of antibiotic resistant bacteria was assessed, revealing that 68% of participants exhibited awareness of the matter. Nevertheless, 49% of participants displayed unfamiliarity with genetically modified organisms (GMOs). When presented with the concept of a UTI solution utilizing genetically engineered bacteria, an encouraging 85% expressed willingness to utilize such a product, demonstrating a favorable public reception to innovative interventions, exemplified by our UTBYE project.

UTI Warriors' Stories

In response to our survey, participants were given the opportunity to share their experiences. Here are some of the responses:

"Many women, like me, have to live in constant fear; as soon as they sense the onset of an infection, it hampers their entire day until they see a doctor and the antibiotics take effect."

"As a result of the infection, I developed kidney complications."

"I have suffered from recurring dreadful infections that required hospitalization and a cocktail of powerful antibiotics."

Why He Was Contacted:

Mr. Lior Porat was specifically contacted due to his role as an Information Specialist. His expertise in literature searches, credibility assessments of sources, and guidance on proper citation practices made him a crucial resource for our research. We sought his assistance to enhance our proficiency in research data utilization, with a focus on optimizing literature searches, understanding source credibility, and navigating various library databases effectively.

What We Learned:

In our session with Mr. Porat, we gained insights into the science of efficient research data utilization. He introduced us to the powerful tool Zotero, unveiling its capabilities for seamless organization and citation management. Through his guidance, we mastered advanced search techniques across diverse library databases, enhancing our ability to retrieve relevant and reliable information. Mr. Porat illuminated the unique subject tree feature of Embase, offering us a novel perspective on refining search queries. His expertise extended to the realm of authenticity verification, equipping us with tools and strategies to discern genuine research articles. Additionally, he demystified the landscape of open access publishing, particularly using platforms like Scopus, enriching our understanding of scholarly dissemination.

The main creibility verification guidelines he provided were as follows:

1. Verify the citation count on Google Scholar.
2. Confirm if the paper is available in the Technion library, ensuring it has been reviewed by Technion experts.
3. Use the JCR database to check the paper's impact factor and ranking. Firstly, verify if it possesses an impact factor. You can also categorize journals to identify top-tier publications.
4. If the journal lacks an Impact Factor, cross-reference it with the Cabells blacklist. If it's listed there, it's not credible.

What We Did:

Inspired by Mr. Porat’s expertise, we put our newfound knowledge into action. Armed with advanced search techniques, we delved into the depths of various databases, retrieving comprehensive and precise data for our research projects, and verified the credibilty of the papers we already used retrospectively in case of low credibility.

Why She Was Contacted:

As the lab manager and a research assistant for our primary PI, Prof. Roee Amit, and with her vast experience, Dr. Goldberg was an invaluable resource for guiding our team in the initial lab setup and procedures.

What We Learned:

We gained insights into effective lab practices, experiment planning, the importance of meticulous organization and preparation, ensuring safety in the lab environment, and the best practices for organizing our storage and equipment. Advised on cloning and other synthetic biology processces.

What We Did:

Under Dr. Goldberg's guidance, we set up our lab workspace, sourced necessary materials and equipment, and devised a structured plan for our experiments.

Why She Was Contacted:

We reached out to Zeinat for her extensive knowledge in women's health and her innovative approach stemming from her iGEM experience.

What We Learned:

Our conversation underscored the urgency for effective and lasting medical solutions for conditions mainly affecting women, such as rUTIs.

What We Did:

We leveraged the insights from our discussion with Zeinat to deepen our understanding of the intricacies in women's health and to steer our subsequent efforts in this area.

Why She Was Contacted:

With her distinct role as Lab Manager under Prof. Sima Yaron and her experience with Lactobacillus Crispatus, Dr. Dina was identified as the right expert to consult on the intricate aspects of growing this specific bacteria.

What We Learned:

Dr. Sahar provided us with invaluable guidance on utilizing the 'jara' for creating low oxygen environments conducive for Lactobacillus Crispatus. She generously shared antibiotics, materials, and taught us the nuances of setting up the CO2 bag, ensuring the bacteria thrived in optimal conditions. Moreover, her insights on Bacillus subtilis were also enlightening and helped us in that axis as well.

What We Did:

Armed with Dr. Shahar's guidance, we successfully set up the environment for Lactobacillus Crispatus growth, incorporating her recommendations and utilizing the materials she provided to enhance our research outcomes.

Why He Was Contacted:

Our initial struggles with the L. Crispatus transformation using his protocol led us to seek his expertise to enhance our methodology. You can find the protocol here.

What We Learned:

Working with L. Crispatus is intricate due to its problematic nature. To successfully transform the bacterium, one should focus on reaching an optimal OD of 0.3, indicating the bacterium's exponential growth phase. Furthermore, the use of a smaller plasmid around 2kb is recommended, and the amount of glycine added plays a crucial role in weakening the cell wall, allowing for successful transformation.

What We Did:

In light of the guidance from Prof. Saris, we carried out a subsequent transformation experiment, varying the glycine concentrations to identify the optimal condition. This approach aimed to better understand the bacterium's behavior and refine our transformation process. You can find more about this in the "Results" page.

Why He Was Contacted:

Our initial attempts using a past iGEM team’s protocol for Bacillus subtilis transformation were unsuccessful, prompting us to seek Dr. Shulami's expertise, given her extensive work with this bacterium.

What We Learned:

Dr. Shulami shared an alternative transformation protocol for Bacillus subtilis, emphasizing the nuances that might have contributed to our previous unsuccessful attempts.

What We Did:

With Dr. Shulami's guidance, we implemented her recommended protocol for Bacillus subtilis transformation. The results were promising as we observed growth, contrasting our initial unsuccessful trials using the iGEM protocol. You can find the protocol here.

Why She Was Contacted:

Given her work with uropathogenic E. coli, we approached Dr. Lukačišinová for a potential collaboration on a competition assay, intending to utilize their UPEC strain facilities in conjunction with our engineered Bacillus.

What We Learned:

Dr. Lukačišinová provided insights into the intricacies of working with UPEC strains and offered collaboration possibilities for the proposed competition assay.

What We Did:

While we gained valuable insights from Dr. Lukačišinová and had the potential for full cooperation from her and Prof. Kishony's lab, we determined that there wasn't sufficient time to execute the collaboration within the iGEM scope.

Why She Was Contacted:

We sought Shenav's expertise because of the challenges we encountered with Bacillus subtilis, particularly regarding contamination and identification.

What We Learned:

We gained a deeper understanding of best practices for working with Bacillus subtilis, including bench cleaning, incubation methods, and bacterial identification.

What We Did:

With Shenav's guidance, we made necessary adjustments in our lab practices, adopted her protocols, and attempted a different colony PCR protocol for subtilis.

Why He Was Contacted:

Given Limor’s extensive experience in the pharmaceutical realm and her understanding of the regulatory aspects, especially related to GMOs, she was approached to provide insights that could guide our product development and regulatory navigation.

What We Learned:

• The rigorous requirements for the approval of a new drug, which includes clinical trials and the demonstration of both safety and efficacy.
• The distinct difference between drug regulation and food supplement regulation.
• The ongoing challenges in antibiotic development, notably due to bacterial resistance.
• The significance of a market analysis to ascertain demand and decide on new drug developments.
• An insight into gender considerations in pharmaceutical research.

What We Did:

Armed with Limor's insights, we re-evaluated our approach to GMO-based drug development, factoring in the regulatory challenges and the market need. We also began an exploratory study on potential gender biases in our research methods and considered the prospect of our drug being classified as a food supplement, given its simpler regulatory pathway.