Exploring Bacteriophage/Biofilm Interaction to Control Urinary Tract Infections with Engineered Bacteriophages

Exploring Bacteriophage/Biofilm Interaction to Control Urinary Tract Infections with Engineered Bacteriophages

Most Affected Population

Clinical and Social Impact

Major Clinical and Scientific Problem

Disproportionately affected population:

UTIs have a strong negative impact on patients’ lives, seriously impairing their social and intimate relationships and leading to a significantly decreased quality of life.

The problem in question concerns urinary tract infections (UTIs). UTIs are a major global health concern, ranking just behind respiratory and gastrointestinal infections in prevalence [1].

Women in Adulthood

They occur when bacteria invade and multiply within the urinary system, Depending on the site of infection, UTIs are classified as urethritis (urethra), cystitis (bladder), or pyelonephritis (kidney) [1].

Women in the Peri and Post-Menopausal Period

Global Impact

Health and Mental Burden

Elderly Men

The presence of these bacteria in the urine, also known as bacteriuria, may occur without clinical symptoms or be associated with active urinary tract infection, when it triggers an inflammatory response and signs such as burning, pain, or fever [1].

-----------------------------------------------------------------------

-----------------------------------------------------------------------

Patients Undergoing Catheterization

Clinical Consequences

In most cases, UTIs arise from the ascension of bacteria from the perineal region to the bladder, with Escherichia coli being the main etiological agent [1].

People with Chronic Illnesses

Antimicrobial Resistance

Mestrado em Engenharia Biomédica - Área de Especialização em Biomateriais, Reabilitação e Biomecânica

Elaborado por:pg55659 Inês Gonçalvespg55711 Leonor Guimarães pg55665 Mariana Silva pg56150 Pedro Ribeiro

*Click + for more information*

*Click + for more information*

Exploring Bacteriophage/Biofilm Interaction to Control Urinary Tract Infections with Engineered Bacteriophages

Urgency for Alternative Therapies

Standard Treatment

Proposed Solution

Bacteriophages are natural bacterial predators that infect and lyse specific bacterial strains without disturbing the surrounding microbiota.

Antibiotics

For Uncomplicated Cases:

Continuous Antibiotics Use

In E. coli urinary tract infections, bacteria can invade bladder epithelial cells and form intracellular biofilm-like communities that resist antibiotics and the immune system, often causing recurrent infections.

• Nitrofurantoin;
• Trimethoprim-sulfamethoxazole;
• Fosfomycin;
• Pivmecillinam commonly [1].

This project aims to isolate and engineer lytic phages capable of targeting these persistent bacterial forms.

Antibiotic Resistence

Disrupted Microbiota

For Severe Cases:

By equipping selected phages with reporter genes and anti-biofilm effectors, the goal is to enhance their therapeutic potential as adjuncts to antibiotics, improving infection control and reducing mortality.

• Fluoroquinolones;
• β-lactams;
• Carbapenems [1].

Bacterial Biofilms and Intracellular Resevoirs

Persistent and Recurrent Infections

The proposed work in this solution involves two tasks.

Why is the proposed solution better?

Other Solutions

◦Targets only disease-causing bacteria◦Preserves healthy microbiota◦Replicates at the infection site◦Penetrates biofilms and intracellular

bacterial reservoirs

◦Works alongside antibiotics◦Reduces mortality in resistant infections

-----------------------------------------------------------------------

-----------------------------------------------------------------------

• Vaccines [12];
• Probiotics [13];
• Antimicrobial peptides (AMPs) [2].

Task 1

Limited the Effectiveness of Standard Drugs

Pathogens Shielded from Treatment

Task 2

While promising, these strategies have not yet been adopted as standard clinical practice.

*Click + for more information*

Severe Complications

• Higher treatment failure;
• Greater healthcare costs;
• Increases mortality.

Exploring Bacteriophage/Biofilm Interaction to Control Urinary Tract Infections with Engineered Bacteriophages

References

[1] Mohamed, A. A., El-Zayat, E. M., & El-Shibiny, A. (2025). Efficacy of phage vB_Ps_ZCPS13 in controlling Pan-drug-resistant Pseudomonas aeruginosa from urinary tract infections (UTIs) and eradicating biofilms from urinary catheters. Virology journal, 22(1), 236. https://doi.org/10.1186/s12985-025-02848-x

[2] Mancuso, G., Trinchera, M., Midiri, A., Zummo, S., Vitale, G., & Biondo, C. (2024). Novel Antimicrobial Approaches to Combat Bacterial Biofilms Associated with Urinary Tract Infec tions. Antibiotics (Basel, Switzerland), 13(2), 154. https://doi.org/10.3390/antibiotics13020154

[3] He, Y., Zhao, J., Wang, L., Han, C., Yan, R., Zhu, P., Qian, T., Yu, S., Zhu, X., & He, W. (2025). Epidemiological trends and predictions of urinary tract infections in the global burden of disease study 2021. Scientific reports, 15(1), 4702. https://doi.org/10.1038/s41598-025-89240-5

[4] Al-Anany, A. M., Hooey, P. B., Cook, J. D., Burrows, L. L., Martyniuk, J., Hynes, A. P., & German, G. J. (2023). Phage Therapy in the Management of Urinary Tract Infections: A Comprehensive Systematic Review. PHAGE (New Rochelle, N.Y.), 4(3), 112–127. https://doi.org/10.1089/phage.2023.0024

[5] Dienye, P. O., & Gbeneol, P. K. (2011). Contraception as a risk factor for urinary tract infection in Port Harcourt, Nigeria: A case control study. African Journal of Primary Health Care & Family Medicine, 3(1), 207. https://doi.org/10.4102/phcfm.v3i1.207

[6] Mera-Lojano, L. D., Mejía-Contreras, L. A., Cajas-Velásquez, S. M., & Guarderas-Muñoz, S. J. (2023). Prevalencia y factores de riesgo de infección del tracto urinario en embarazadas [Prevalence and risk factors of urinary tract infection in pregnant women]. Revista medica del Instituto Mexicano del Seguro Social, 61(5), 590–596. https://doi.org/10.5281/zenodo.8316437

[7] Raz R. (2011). Urinary tract infection in postmenopausal women. Korean journal of urology, 52(12), 801–808. https://doi.org/10.4111/kju.2011.52.12.801

[8] Nicolle L. E. (2016). Urinary Tract Infections in the Older Adult. Clinics in geriatric medicine, 32(3), 523–538. https://doi.org/10.1016/j.cger.2016.03.002

[9] Brusch, J. L. (2025, August 20). Catheter-Related Urinary Tract Infection (UTI): Overview. In Medscape. Available from https://emedicine.medscape.com/article/2040035-overview?form=fpf

[10] Ahmed, A. E., Abdelkarim, S., Zenida, M., Baiti, M. A. H., Alhazmi, A. A. Y., Alfaifi, B. A. H., Majrabi, R. Q. M., Khormi, N. Q. M., Hakami, A. A. A., Alqaari, R. A. M., Alhasani, R. A., Alajam, R. A., Alshehri, M. M., Alenazi, A. M., Alqahtani, B., Alshamrani, M., Alhowimel, A., & Abdelwahab, S. I. (2023). Prevalence and Associated Risk Factors of Urinary Tract Infection among Diabetic Patients: A Cross-Sectional Study. Healthcare (Basel, Switzerland), 11(6), 861. https://doi.org/10.3390/healthcare11060861

[11] Mayo Clinic. (2025, September 26). Urinary tract infection (UTI) - Diagnosis and treatment. Mayo Clinic. Available from https://www.mayoclinic.org/diseases-conditions/urinary-tract-infection/diagnosis-treatment/drc-20353453

[12] Nickel, J. C., Kelly, K. L., Griffin, A., Elterman, D., Clark-Pereira, J., & Doiron, R. C. (2024). MV140 sublingual vaccine reduces recurrent urinary tract infection in women Results from the first North American clinical experience study. Canadian Urological Association journal = Journal de l'Association des urologues du Canada, 18(2), 25–31. https://doi.org/10.5489/cuaj.8455

[13] Pastuszka, A., Tobor, S., Łoniewski, I., Wierzbicka-Woś, A., Sielatycka, K., Styburski, D., Cembrowska-Lech, D., Koszutski, T., Kurowicz, M., Korlacka, K., Podkówka, A., Lemiński, A., Brodkiewicz, A., Hyla-Klekot, L., & Skonieczna-Żydecka, K. (2025). Rewriting the urinary tract paradigm: the urobiome as a gatekeeper of host defense. Molecular biology reports, 52(1), 497. https://doi.org/10.1007/s11033-025-10609-w

Patients Undergoing Catheterization

Present a high risk (between 90 and 100% of patients with long-term catheterization develop bacteriuria) [9].

Task 1: Isolation, Characterization, and Selection of Phages

Women in Adulthood

Due to anatomical factors (short urethra and proximity to the anus), it is estimated that 50–60% of women will have at least one episode in their lifetime [4]. The incidence is high among young, sexually active women, particularly among contraceptive users (prevalence of 71,9%) [5]. During pregnancy, the risk increases, with a prevalence of 37,7%, and is also associated with maternal and fetal complications [6].

Task 2: Phage Engineering with Reporter Genes and Effectors

Different engineering strategies will be used:

(i) rebooting of in vitro-assembled phage genomes (for smaller genomes)(ii) CRISPR-Cas-9-assisted editing (for larger genomes)

People with Chronic Illnesses

People with chronic diseases, such as diabetes, are at greater risk of UTIs. In patients with type 2 diabetes, the prevalence is 25,3%, being higher in women (41,1%) than in men (7,2%) [10}.

Health and Mental Burden

UTIs not only impose a considerable economic burden but also disrupt daily life and increase psychological stress [3]. More than half of UTI patients experience mental health challenges, including anxiety and depression. Consequently, UTIs represent a growing and serious public health concern [3].

Antimicrobial Resistence

The high prevalence and recurrence of UTIs often result in frequent and sometimes prolonged antibiotic use, which drives the selection of resistant strains, particularly of Escherichia coli [3]. This trend poses an increasing challenge for clinical practice, narrowing therapeutic options and raising the risk of treatment failure, thereby contributing to the global problem of antimicrobial resistance [3].

Women in Peri and Post-Menopausal Periodal

The frequency of UTIs increases due to hormonal changes, such as reduced estrogen and changes in vaginal flora. The prevalence of bacteriuria is 15-20% between the ages of 65 and 70, reaching 20-50% in women over 80 [7].

Global Impact

UTIs are among the most common infectious diseases worldwide, affecting about 150 million people annually [2].They cause significant morbidity and economic burden, with reaching around $3,5 billion in societal costs [2].UTIs were linked to around 260,000 deaths in 2019. Mortality increases progressively with advancing age, peaking in the elderly population, as shown in the graph on the right [3].

* Age-standardized rate adjusts for differences in age distribution, allowing fair comparison of rates (e.g., disease or mortality) between populations or over time.

Elderly Man

Due to frailty, hormonal changes, comorbidities and increased ose of catheters, they also have a high frequency of UTIs [8].

Clinical Consequences

The impact of UTIs extends beyond discomfort and social repercussions: they may lead to severe clinical consequences, including pyelonephritis, sepsis, and, in the most critical cases, multiorgan failure. Such complications significantly increase both mortality and hospitalization rates [3].