Periodic Reporting for period 1 - URISAMP (Disease and vaccine monitoring based on non-invasive genital tract liquid biopsy sampling.)
Reporting period: 2022-05-01 to 2024-10-31
The ability to easily obtain appropriate and reliable biological samples is critical to advance life sciences and public/personal health. Having a sample that is non-invasive and can be self-collected at home has major acceptance and feasibility advantages. Our team has already demonstrated that first-void urine (FVU) samples can replace clinician-collected cervical samples in Human Papillomavirus (HPV) based cervical cancer screening. Using the initial or first-void urine stream is crucial here. Indeed, the outer layers of the epithelium of the genital tract exfoliate and together with the genital mucus form the utero-cervico-vaginal secretions. This discharge accumulates between the labia minora, around the urethral opening, and is washed away with the FVU upon initiation of urination. The URISAMP project uses HPV infection and vaccination as a model to prove the hypothesis that FVU is an ideal genital tract liquid biopsy to monitor both disease and immune response for women at the site of infection.
The specific objectives of URISAMP are to:
• Identify and validate analytical methods to detect relatively low and fluctuating concentrations of potential biomarkers in FVU.
• Provide and validate high-throughput protocols to study neutralizing antibodies in FVU.
• Test the developed assays’ robustness in well selected cohorts.
• Validate the use of FVU to monitor natural and vaccine-induced anti-HPV antibodies.
• Demonstrate interaction between vaccine-induced anti-HPV antibodies and HPV virions in women with a productive infection.
The specific objectives of URISAMP are to:
• Identify and validate analytical methods to detect relatively low and fluctuating concentrations of potential biomarkers in FVU.
• Provide and validate high-throughput protocols to study neutralizing antibodies in FVU.
• Test the developed assays’ robustness in well selected cohorts.
• Validate the use of FVU to monitor natural and vaccine-induced anti-HPV antibodies.
• Demonstrate interaction between vaccine-induced anti-HPV antibodies and HPV virions in women with a productive infection.
Within the URISAMP project, we have successfully been working on:
• Validating FVU as sample to monitor natural and vaccine-induced antibodies.
• Optimizing protocols for detection of neutralizing anti-HPV antibodies in FVU samples.
• Producing HPV pseudovirions for the nine HPV genotypes included in the nonavalent HPV vaccine.
• Producing HPV genotype-specific monoclonal antibodies, using our in-house produced pseudovirions as antigens.
• Developing and validating immunoassays for the robust detection of genotype-specific anti-HPV antibodies.
• Evaluating and optimizing HPV virion isolation protocols.
• Establishing a clinical trial to collect samples for confirmation of robustness and sensitivity of the developed anti-HPV immunoassays and to examine antibody fluctuation patterns and biomarkers to normalize these fluctuations.
• Setting-up a clinical trial to collect samples for demonstration of interaction between anti-HPV antibodies and HPV pseudovirions or wild-type virus particles.
• Evaluating FVU as a sample for vaginal microbiome analysis, assessing the potential effect of the vaginal microbiome on HPV infections.
• Evaluating FVU as sample to monitor local cellular immune responses to HPV.
• Assessing the potential of FVU to explore the immune response to other genital tract infections.
The URISAMP project pioneers several novel methodologies that are expected to make a significant impact in the field of vaccinology and monitoring of genital tract infections. Up to now, our work and publications have had profound impact on HPV vaccine monitoring and related research, establishing us as a leading expert centre worldwide in the use of first-void urine for HPV-related research.
• Validating FVU as sample to monitor natural and vaccine-induced antibodies.
• Optimizing protocols for detection of neutralizing anti-HPV antibodies in FVU samples.
• Producing HPV pseudovirions for the nine HPV genotypes included in the nonavalent HPV vaccine.
• Producing HPV genotype-specific monoclonal antibodies, using our in-house produced pseudovirions as antigens.
• Developing and validating immunoassays for the robust detection of genotype-specific anti-HPV antibodies.
• Evaluating and optimizing HPV virion isolation protocols.
• Establishing a clinical trial to collect samples for confirmation of robustness and sensitivity of the developed anti-HPV immunoassays and to examine antibody fluctuation patterns and biomarkers to normalize these fluctuations.
• Setting-up a clinical trial to collect samples for demonstration of interaction between anti-HPV antibodies and HPV pseudovirions or wild-type virus particles.
• Evaluating FVU as a sample for vaginal microbiome analysis, assessing the potential effect of the vaginal microbiome on HPV infections.
• Evaluating FVU as sample to monitor local cellular immune responses to HPV.
• Assessing the potential of FVU to explore the immune response to other genital tract infections.
The URISAMP project pioneers several novel methodologies that are expected to make a significant impact in the field of vaccinology and monitoring of genital tract infections. Up to now, our work and publications have had profound impact on HPV vaccine monitoring and related research, establishing us as a leading expert centre worldwide in the use of first-void urine for HPV-related research.
Several of our project’s significant achievements can be considered breakthroughs, advancing the research field significantly beyond the state of the art.
With URISAMP, we are the first to report the detection of vaccine and naturally induced antibodies in non-invasively collected first-void urine, marking a significant advancement in tools for understanding local immune responses in the female genital tract. We demonstrated that anti-HPV antibodies can be detected in up to 100% of FVU samples post-vaccination and remain detectable for up to 12 years, with FVU HPV16-specific IgG results showing good correlations to paired serum. Hence, we have sparked considerable interest in using these non-invasive samples for monitoring humoral immune response in larger studies.
Moreover, recently, we published compelling data that antibodies in FVU samples remain neutralizing, supporting the hypothesis that vaccination at a later age might reduce autoinoculation rates and impact transmission to sexual partners.
The optimized methods for immune response detection and monitoring based on FVU, developed in the URISAMP project, will play an important role in future (long-term) follow up of HPV vaccination trials, epidemiological/large cohort studies and studying the natural history of an HPV infection. Moreover, if URISAMP proves successful, and FVU can also replace other sample types (e.g. blood), applications will largely extend the STI field, and its impact as a novel diagnostic and prognostic tool for health assessment will be substantial.
With URISAMP, we are the first to report the detection of vaccine and naturally induced antibodies in non-invasively collected first-void urine, marking a significant advancement in tools for understanding local immune responses in the female genital tract. We demonstrated that anti-HPV antibodies can be detected in up to 100% of FVU samples post-vaccination and remain detectable for up to 12 years, with FVU HPV16-specific IgG results showing good correlations to paired serum. Hence, we have sparked considerable interest in using these non-invasive samples for monitoring humoral immune response in larger studies.
Moreover, recently, we published compelling data that antibodies in FVU samples remain neutralizing, supporting the hypothesis that vaccination at a later age might reduce autoinoculation rates and impact transmission to sexual partners.
The optimized methods for immune response detection and monitoring based on FVU, developed in the URISAMP project, will play an important role in future (long-term) follow up of HPV vaccination trials, epidemiological/large cohort studies and studying the natural history of an HPV infection. Moreover, if URISAMP proves successful, and FVU can also replace other sample types (e.g. blood), applications will largely extend the STI field, and its impact as a novel diagnostic and prognostic tool for health assessment will be substantial.