Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - PROM (Pregnancy and Reproduction Organ-on-a-chip Models)

Teaser

PreTerm Birth (PTB) occurs before 37 weeks of gestation. Of 60,000 babies born prematurely every year in UK, ~1,400 die. Premature babies show increased risks of long term neurodevelopmental deficits and chronic diseases.Preterm Rupture of the foetal Membranes (PROM) is a...

Summary

PreTerm Birth (PTB) occurs before 37 weeks of gestation. Of 60,000 babies born prematurely every year in UK, ~1,400 die. Premature babies show increased risks of long term neurodevelopmental deficits and chronic diseases.
Preterm Rupture of the foetal Membranes (PROM) is a syndrome attributable to multiple factor. Evidences linked environmental toxicant exposure and infection with a decreased fertility and an increase PTB rate.
Studies of these topics suffered for limits of in vitro cell culture (e.g. absence of extracellular environment, dilution) and anatomical, physiological, endocrine differences between human and animal models.
Long term goal of this project is to create Organ-on-a-Chip (OoC) models of the human endometrium and the human foetal membranes to assess the effects of environmental insults, such as infection or inflammation, to exacerbate preterm birth.
An overarching goal is to utilize these OoC models to identify personalized therapy to improve and preserve fertility.

Thanks to this fellowship, I thus designed and developed 2 microfluidic systems that allow to grow human, patient-derived cell of the uterus and the foetal membranes in a controlled environment to recreate their natural physiological functions.
These devices have been designed with inputs from experts in the field of microfluidics, infectious diseases and reproductive biology and toxicology.
In parallel I focused on early embryo development to design a new device for in vitro fertilization.
The results of this research have been presented at several international conferences and have been summarized in 1 PCT patent, 3 published papers and 2 papers in preparation.
In these 2 years, I have learnt analytical methods (qPCR, ELISA, Mass Spectrometry, proteomics, metabolomics) and tissue culture techniques (embryo culture and cell isolation protocols) that were not part of my engineering background. I also established new collaborations in the field of manufacturing, sustainable plastics and tissue engineering, to define new approaches for the fabrication of the new generation of organs-on-a-chip and for the integration of biocompatible scaffold in the microfluidic device.

Work performed

Thanks to my Marie Curie Fellowship, I was able to transition from my position as Research Assistant Professor at Vanderbilt University (VU) to University Academic Fellow (UAF) at the University of Leeds (UNIVLEEDS) to establish my independent career in the field of microfluidics and reproductive toxicology.
After working as Co-Investigator in large US projects to design new organs-on-a-chip models and collaborating with world experts in reproduction and infectious diseases, I defined my academic career plans with the aim to create innovative technologies to support fertility treatments as well as to identify the causes and origins of miscarriage and preterm birth (“reproductive technologies”). During this project:
• I increased my publications track record (31 papers, max cit. 154);
• I established active collaborations in embryology and fertility at UNIVLEEDS (e.g. H.Picton, J.Walker, N.Forde, N.Orsi).
• I received International recognition (EAMC-2017 International Association of Advanced Materials - Scientist Medal, SRI-2019 Training Investigator Poster Award).
• I have been invited to national and international conferences
• I attracted funding for my research: NC3Rs-MRC EASE-CRACK IT “Design, Fabrication and Testing of a Mouse Embryo Culture Chip” (Principal Investigator), MRC Confidence in Concept “Integration of nanosensing in microfluidics to improve success of fertility treatments”, EPSRC TTL Organ-on-a-chip Network sabbatical awards, Marie Skłodowska-Curie Innovative Training Network “SENTINEL-Single-Entity NanoElectrochemistry”, Grow Med Tech Proof of Concept “Microfluidic embryo culture as a device to improve the efficiency of infertility treatments in humans”, EPSRC IAA “Compatibility of time-lapse Imaging and Nanosensor Integration with Microfluidic Culture of Embryos as A means To Improve The Efficacy Of Human Infertility Treatment”.
These enabled me to set up and support my research group (4 PhD students, 1 PostDoc), to inspire them and other students (>20 PhD and Master Students supervised in the past) to creatively work and translate ideas into entrepreneurial activities by generating IP and filing new patents as starting point to develop new business ideas (two UK patents describe the technology developed during this fellowship and one is now a PCT application).
• I collaborated with world leaders, such as:
-Prof. D. Aronoff, Vanderbilt University, USA, Prof. McLean at the Centre for Innovative Technology, Vanderbilt University, USA, in the fields of infective diseases and metabolomics,
-Prof. Helen Picton, UNIVLEEDS in the fields of embryology and Oncofertility.
and accessed the cutting-edge microfabrication technologies available at UNIVLEEDS and national and international leading research centres (Heriot-Watt University in Edinburgh, Vanderbilt University, Nashville, USA).

Final results

During this fellowship I developed two microfluidic devices that model the foetal membranes and the endometrium, using human, patients- derived cells. These systems represent an innovative, powerful tool to understand specific mechanisms and functions of the female reproductive organs there could not be studied before with animal models or traditional in vitro culture.
I developed these systems in collaboration with Vanderbilt University and adapted designs later on at the University of Leeds after learning about cells proliferation, effects of substrates and manufacturing challenges.
These systems are now ready to be distributed to collaborators and to be used in Leeds by my team for observing chorionamnionitis and bacterial infection in the amniotic sac and for completing toxicology studies on the endometrium.
These two platforms have a tremendous impact on our understanding of the pathophysiology of significant diseases such as endometriosis, and of the mechanisms of preterm birth and of the foetal maternal interface equilibrium.
In addition to this, thanks to collected results, opportunities for networking and skills gained during this fellowship, I have been able to design a new, safe, and easy-to-use microfluidic system that has the potential to improve the success of In Vitro Fertilization (IVF) treatments. This device has been patented, tested with murine and bovine embryos to confirm the proof of concept and to validate our concept, and it is now being translated in commercial product. Results obtained so far confirmed these benefits in animal studies, and showed the advantage of using this system to culture embryo in vitro. This system thus represents a new, useful tool for research since it simplifies the IVF process in animals (mice and bovine tested so far), reduces the costs, time and invasiveness of the procedure (as required by NC3Rs sponsorship) and allows to control, monitor and challenge the embryo during its development. Moving towards the human IVF market, this device will be manufactured and distributed through a new company starting at Leeds in the coming year, will complete the preclinical testing phase to then start human clinical trial. If successful as planned, the device will change the IVF procedure, make it more easy, more consistent, increase success rate (currently below 30%) and thus decrease costs for patients and health care system in UK, Europe and worldwide.

Website & more info

More info: https://engineering.leeds.ac.uk/staff/870/Dr_Virginia_Pensabene.