Esophageal cancer

In our project, we focused on the early detection of esophageal cancer since it is one of the most fatal malignancies worldwide, with a dramatic increase in incidence in the Western world occurring over the past few decades. Moreover, it is often diagnosed during advanced stages, because of the lack of early clinical symptoms [1]. Because of its poor prognosis it is the sixth most common cause of cancer-related death [2]. 5-year survival rates have been shown at rates of 10–15% [3].

We are confidient that the number of deaths caused by esophageal cancer can be reduced by early detection.

Talk with a general practitioner

To learn more about the medical necessity we had a conversation with Dr.med. Jörg Meckoni. Through his years of experience as a general practitioner, he was able to give us insight into his way of dealing with esophagus-like symptoms. He told us about the very non-specific symptoms. These include difficulty swallowing and loss of appetite. As a general practitioner, the first step in such symptoms is to treat the symptoms, but there is usually no investigation of the cause or testing for esophageal cancer. Testing for esophageal cancer is recommended only if the patient exhibits typical risk behaviors or preexisting conditions. These include smoking, alcohol consumption and chronic reflux. If there is no improvement after a longer period of time, the patient is referred to the specialist.

“The poor prognosis and increasing incidence of esophageal cancer highlight the importance of improved detection and prediction methods prior to treatment.” - Jörg Meckoni, general practitioner

He also told us more about the current detection methods of esophageal cancer. These include different screening/diagnostic tests e.g. oesophagogram and endoscopic ultrasound (EUS) with computerised tomography scan and/or positron-emission tomography scan to determine tumour metastasis.

Shortcomings of these screening/diagnostic tests include:

• low sensitivity
• high cost
• time-consuming procedures
• invasiveness and risk of bodily exposure to radioactive substances
• requirement for the services of an expert

“To improve the diagnosis of esophageal cancer, it is crucial to look for novel non-invasive blood-based diagnostic.” - Jörg Meckoni, general practitioner

Proposed solution: Lion Detect

Lion Detect - A fast, new generation of a detection system that can detect even low antigen concentrations with the help of signal amplification. It is a system that is universally applicable for the detection of various antigens and offers the possibility for continuous further development. We have committed to providing easy and cost-effective detection of esophageal cancer directly in the doctor's office through a new rapid test. This should enable patients to be treated more quickly and thus increase the chances of successful treatment.

Integrated human practice

In order to develop an innovative detection system that can be used in the doctor’s office, we have been in constant contact with experts. With their feedback, our project could be adapted and improved on several levels.

Talks with experts to create the perfect three components

After we had formulated our idea, we started directly with the exchange with experts.

Detection component: With the idea of detecting the antigens with antibodies, we turned to Prof. Dr. Steffan Dübel. He is an expert in the field of antibodies and serves as director of the technology transfer unit "Centre for Molecular Engineering" of iTUBSmbH. Moreover, he initiated the "Antibody factory" of the German National Gemome Research Network and is editor of the four volume "Handbook of Therapeutic Antibodies" and other antibody engineering books. It turned out that the production of complete antibodies in a microorganism is very difficult. Based on his feedback we decided to change our plan and work with ScFv fragments, which are only fragments of an antibody.

Amplification component: For this component, we decided to work with inteins, as they were a potential system that came up during the research. However, there are several splicing mechanisms. In 2020, the iGEM team from Barcelona already worked with inteins, so we decided to meet with a former team member of their team, Andreu Pascuet. We have talked about several variants that allow detection of antigens by means of inteins. Our preferred variant was the one used by the iGEM team barcelona in 2020. However, in the discussion it turned out to be disadvantageous for our project. He recommended to work with split-inteins and pointed out that the interaction should be prevented as long as no antigen is present. Based on his feedback, we continued our research and came across the caged variant of the inteins, which was ultimately also used in our project.

Reporter component: For this component, our plan was to work with astaxanthins, as they represent a red dye that is readily visible in E. coli. Therefore, we turned to Dr. Nadja Henke - an expert in the field of carotenoids. While talking to her, it turned out that astaxanthins are not suitable for our project, because enzymes of the last steps of astaxanthin biosynthesis are membrane-bound. As we want to work with a cell lysate, astaxanthins were no longer an option. This feedback led us to reconsider our plan of the reporter component. In the end, we decided to use a common and well known reporter mechanism - Förster resonance energy transfer (FRET).

"Flourolion" box for the use in the doctor's office

After we had gone through the first engineering cycle in creating the Flourolion box we had a conversation with the medical doctor Dr. Christos pantazis. Through his own working experience, he told us that our box would only establish itself if the detection could be done in a few steps. Based on this feedback, we went through the second engineering cycle of creating a better version of the Flourolion box and could therefore the individual pipetting steps could be replaced with an automatic pumping of the lysates into the box.

Trip to IDT-Biologika

In order to broaden our horizon, we got the opportunity to have a look at the research and development of IDT Biologika. Besides a very interesting trip to the labs, we got the opportunity to present our project in front of Dr. Simone Kardinahl. She is a strategy and goal orientated production leader with 20 years of experience along the entire value chain in the global pharmaceutical industry. She pointed out that we should take a closer look at the competitive situation of our product. This led us to compare the price of our product with the prices of other systems. Moreover, we adjusted the box in the second engineering cycle to use as little material as possible, trying to lower the price further.

References

1. Huang FL, Yu SJ. Esophageal cancer: Risk factors, genetic association, and treatment. Asian J Surg. 2018;41(3):210-215. doi:10.1016/j.asjsur.2016.10.005
2. Zhang Y. Epidemiology of esophageal cancer. World J Gastroenterol. 2013;19(34):5598-5606. doi:10.3748/wjg.v19.i34.5598
3. Moyana TN, Janoski M. Recent trends in the epidemiology of esophageal cancer. Comparison of epidermoid- and adenocarcinomas. Ann Clin Lab Sci. 1996;26(6):480-486.
4. van der Watt PJ, Okpara MO, Wishart A, et al. Nuclear transport proteins are secreted by cancer cells and identified as potential novel cancer biomarkers. Int J Cancer. 2022;150(2):347-361. doi:10.1002/ijc.33832