To settle on our idea for this year, we followed the aim of the iGEM which is "local people solving local problems". We started looking for a group of people who are affected by preventable diseases to cut the problem from its stem. We focused on groups sharing certain characteristics and features, for example: people who live in the same geographical region or the same city, or people of the same age suffering from a certain illness. Of all the groups on the table, children grabbed the most attention as they are the most vulnerable beings of mankind. They undergo rapid physical and mental development that needs preventative care to meet their biological ad psychological needs. This goes without mentioning our inspiration from Nelson Mandela's quote "Our children are our future, and one of our most basic responsibilities is to care for them in the best and most compassionate manner possible." We started consulting specialists, and we met Dr. Rehab Abdulhai, professor of Public Health & Community Medicine, Cairo University.
She revealed devastating statistics; in developing countries, 50% of deaths occur in children under 5 years old. She also told us that the United Nations General Assembly set a Sustainable Developmental Goal (SDG) to reduce newborn mortality to 12 per 1,000 live births and under-five mortality to 24 per 1,000 live births (SDG 3.2). With all these truths on the table, we set our minds that our project must benefit children by finding new treatments for any common diseases affecting them, improving current medicine, or diagnosing diseases as early as possible to start their treatment immediately and prevent any further deterioration. Here comes the role of synthetic biology to offer us hope for a new treatment option that could cure these upcoming generations (children) and guarantee a better future, health, and quality of life.
A problem that faced us was that tons of diseases affect children and newborns. How were we to choose a specific one for the prototype of our new theranostic(therapeutic and diagnostic) approach? After several meetings and discussions, we concluded that the most preventable diseases and easily treated ones are inborn errors of metabolism. Such a conclusion helped narrow the scope of our quest, by heading to different pediatric clinics looking for the most common inborn errors of metabolism that have a late presentation and poor diagnosis.
We held a meeting with Dr. Radwa Mohamed Azab, assistant professor of pediatrics at Cairo University. She told us about the most prevalent metabolic disorders among children in Egypt, which phenylketonuria was on top with a prevalence rate of 1 in every 5000 live births, & even after being diagnosed, the children deteriorate and face most of the complications of the disease. When asked about her thoughts behind this, she told us that this disease is screened for at birth. However, the results of the screening test come out after 8-12 weeks, which causes high drop-outs. Moreover, hospitals lose contact with parents within this period and so even if diagnosed, the children's parents won't know about it. As for the management, there is low awareness of the dietary needs of Phenylketonuria patients as most parents comply with their children's will and feed them what they want, not what they need.
To see the facts with our own eyes, we visited Al-Galaa pediatrics hospital and Ghamra obstetric hospital to interview future parents.
In the pediatric hospital, we interviewed the Doctors & handed out surveys regarding the inborn errors of metabolism they diagnose the most in their practice. We asked the Doctors to provide 3 answers in descending order, with the most prevalent disease on top and the least on the bottom. Furthermore, we added a scoring system: the most prevalent disease gets 3 points, the middle one gets 2, and the last gets 1.
As expected, phenylketonuria got the highest score. Not only that, but even when patients sought medical advice, it was too late as irreversible consequences had already occurred. Lastly, a doctor who asked about our project recommended adding a label on our chip and writing patients' info on it, to avoid mixing of chips.
As for the obstetric hospital, Ghamra, we interviewed the parents. We asked them about their previous knowledge of inborn errors of metabolism and then specified the question to phenylketonuria. We also enquired about their ideas of possible causes of the disease, and if they know about the relation between such disease and consanguinity.
We started our journey by looking for the best diagnostic tools available. However, there was a huge variety of options and choices in the fields of diagnostics and therapeutics. We decided to start with using microfluidics for our project.
To ensure that we were going on the correct path, we had a meeting with Dr. Mohamed Abdelgawad, a microfluidics expert and professor at Assiut University. He provided us with a great insight into microfluidics and demonstrated the several classes of microfluidic devices including paper, controller flow, and droplet-based microfluidics. He provided us with the pros and cons of using such an approach. However, the disadvantages outweighed the advantages by far as microfluidics would consume a huge budget, and its use is limited in our local community. Still, he recommended using paper microfluidics due to its relatively low cost. However, being responsible for the only available microfluidics fabrication and manufacturing center in Egypt, he demonstrated that no clean room is currently available at the center. Consequently, we should seek the help of other institutes like the American University in Cairo which may provide more help in choosing the proper device for our application.
Following Dr. Mohamed Abdelgawad's advice and giving microfluidics one last chance, we headed to the American University in Cairo (AUC) to meet with Dr. Reda Abdelbaset, Biomedical Engineering professor at the Center of Nano electronics & Devices. He pointed out that to construct a proper microfluidic chip, we need a clean room as the smallest particles of dust may block the minute channels of the chip. Furthermore, he proposed the usage of Printed Circuit Boards (PCBs). However, it will not be the user friendly approach we planned for and it would have a comparatively high cost. In terms of shipping and availability, this item is unfortunately unavailable in our country as no industry for its manufacture is present in Egypt. In this regard, we even tried to proceed with the shipping process. Yet, it was time consuming and required a lot of effort to design the needed chip for the PCBs. Also, the shipping process would take four months to arrive.
As part of our institute's partnership with Michigan State University (MSU), we asked for a meeting with Dr. Eran Andrechek, associate professor in physiology. We presented our idea and our progress and told him that we needed to create a user-friendly diagnostic device for phenylketonuria. He emphasized the idea that microfluidics would not be user friendly. Fortunately, he referred us to using the Lateral Flow Assay technique as it has a low cost, is simple to use, and provides rapid results. Afterward, he explained the technique itself, which is a paper-based platform for the detection and quantification of analytes in complex mixtures, where the sample is placed in a testing device and the results are displayed in a very short time. Lastly, he enlightened our minds to a great idea. The professor had a lifetime complaint of products being called user-friendly although they are not suitable for some special conditions such as color blindness. Accordingly, we decided to consider special conditions when finalizing the kit.
After settling on the Lateral flow assay, we decided to have a meeting with Dr. Fareed Aboul-ela, Associate Professor of Biophysics at Zewail City of science and technology. He explained that lateral flow assay has disadvantages such as low specificity, false-positive and false-negative results when using antibodies. So, we suggested using aptamers, which are short stretches of artificial DNA or RNA that bind to a specific target molecule. Dr. Aboul-ela confirmed that phenylalanine does not have a previously used verified sequence, but the available sequences are now predicted with specific tools and the ideal way to get the best sequence is the Using SELEX, Systematic Evolution of Ligands by Exponential Enrichment. He went on to explain the SELEX steps, which consist of incubation, binding, partitioning, washing, targeted elution, and amplification. In the end, he mentioned that SELEX is not the only way to find the sequence of aptamers and that we can use deep learning technology. He further explained the use of deep learning technology and guided us to use it in our diagnostic tool.
John Maynard Keynes, English economist, once said "the difficulty lies not so much in developing new ideas as in escaping from old ones". This made us think of how to convince the healthcare workers and even the people to shift from their traditional and apparently successful method to our new one. Accordingly, we arranged meetings with experts to help us dodge this hurdle.
Our first meeting was with Dr. Wagida Anwar, Chair Biomedical Research Department at the Armed Forces College of Medicine (AFCM). She pointed out many challenges and obstacles that might face us, but she mainly focused on the idea of convincing the people about the project; to do that, in her opinion, we had to focus on the economic benefit of our device and prove that it is cost-effective, safe to use, and follows their common customs and beliefs. Following her advice, we started with the economic side and changed the items we use from expensive microfluidics types to the cheapest ones, until we deviated from the idea of microfluidics as a whole. Then, we worked on the safe side and integrated the anti-CRISPR technology to ensure that the circuit is 100% safe, and simplified the idea to be explainable to the public. She also lectured us about all the steps regarding ethical issues. She answered our concerns about working with mice cell lines and the process of clinical trials. Lastly, she offered to supervise our progress until our project is finished to ensure that none of the worldwide ethical guidelines is violated.
We then had a meeting with Dr. Nadia Zakahry, the Egyptian former Minister of Scientific Research. She pointed out the difficulty of applying our device in clinical settings and convincing the healthcare workers about it, which lead to our planning to ensure high safety, higher sensitivity, and specificity. Finally, she warned us about the greatest obstacle which is funding. She pointed out that many great projects and ideas are put off and not converted to real items simply due to funding issues and that’s why we would need financial support and to contact stakeholders. As a result, we made a complete entrepreneurship model for our project.
