Need and Challenge

Cancer is one of the highest-ranked causes of death in the world. According to the WHO database (WHO, 2022), almost 10 million people will die because of cancer in 2020. Female breast cancer (2.26 million cases) was the most diagnosed cancer, followed by lung (2.21 M) and colon and rectum (1.93 M). Meanwhile, lung cancer (1.79 million deaths) was the most common cause of death among all cancers, before colon and rectum (916 000) and liver cancers (830 000) (Ferlay, J., et al., 2021).

Genetic changes can cause cancer, through errors in cell division, DNA damage caused by harmful substances, or inheritance (NIH, 2021). Enormous amounts of DNA damage lead to mutation, and hence the cell becomes abnormal and malfunctioning, causing extreme proliferation and forming tumors. Reports (CDC, 2017) (Li, X., et al., 2018) (Sharp, L., et al., 2014) show that the incidence of cancer is higher in urban places like the USA, Shanghai, and Ireland. Some substances can be harmful to our DNA, especially when we live in urban environments where processed products and chemicals are ubiquitous.

The EU Science Hub (EU, n.d.) states that there is a lack of tests available for the assessment of carcinogens. The two-year bioassay study in rodents acknowledged by the OECD is adequate to predict cancer risk in humans, however, is doubtful with notable challenges and uncertainty associated with extrapolating from rodents to humans, with quantitative risk estimation and limited accuracy. In such a case, a crucial requirement emerges for developing a reliable carcinogen detection method aimed at identifying detrimental carcinogens around us with less time, resources, and animal use. We will attempt to design a novel biosensing system.

Governments and companies can employ our innovative method to test their products, thereby eliminating harmful substances that can cause DNA damage before their sale or implementation. We understand that the monetization and implementation of our solution has long-term implications; adopting this proactive approach will significantly reduce the likelihood of individuals being exposed to carcinogens, and subsequently lessen the burden on the healthcare system, contributing to the achievement of the third SDG, good health and well-being. By minimizing the burden of cancer on the healthcare system, medical resources can be re-allocated to other branches of the system, and less medical waste will be produced, therefore contributing to SDG 11 and 12: Sustainable cities and communities, and responsible consumption and production.

Market analysis

Market analysis reveals that cancer, the leading cause of morbidity and mortality worldwide, presents a significant opportunity for businesses in the healthcare industry. According to the World Health Organization (WHO), cancer is responsible for millions of deaths annually, resulting in a growing demand for effective solutions. Factors such as aging populations, lifestyle changes, improved detection, and diagnosis contribute to higher cancer incidence rates in high-income countries. However, low- and middle-income countries are also witnessing an alarming increase in cancer rates due to population growth, urbanization, and the adoption of unhealthy behaviors.

The rising awareness of the detrimental effects of carcinogens on human health has fueled the demand for carcinogen detectors. These devices play a crucial role in identifying and measuring potential cancer-causing agents, enabling individuals and organizations to take preventive measures and reduce exposure risks.

To conduct further investigation of the market, we reached out to GENENET Technology at Stevenage and Moderna. The founders of the company evaluated our project as a highly challenging one that requires advanced technical expertise and involves extensive data analysis andinterpretation. They recognized the potential of our design in the market, considering it highly innovative and cutting-edge in the fields of biology and chemistry, areas that are constantly evolving with frontier science and technology.

With that being said, they suggested that we use SWOT analysis to clarify the market value, market positioning, and other advantages of introducing our bio-sensor. At the same time, they point out that developing and launching a biosensor can require significant financial resources. We may face challenges in securing adequate funding for research anddevelopment, manufacturing, marketing, and distribution of yourproduct. Therefore, with limited entrepreneurial resources and experience, we tried to explore various funding options such as grants, investors, crowdfunding, or partnerships to overcome the obstacles.

Scientific Evidence and Technical Approach

We tested the RecA-eGFP construct with UVA, UVB, H2O2, Nalidixic acid, and Aspartame. The RecA promoter is involved in the SOS response of E. coli, and it will be activated if the DNA is damaged. This design was invented and optimized by team BIT 2019, which demonstrated its ability to report DNA damage caused by H2O2. Our results using cell imaging and plate readers showed that the number of significant fluorescence units increases when the concentration or intensity of the DNA damage-inducing agents rises.

Next, we also successfully transfected two vectors of our FRET design into HEK293 cell lines and validated the FRET of ATRIP and RPA1 after UVB treatment (Refer to New Parts). In future stages, we will test our FRET system with the carcinogens mentioned above.

Innovation

Current solutions: Comet assay

The Comet Assay, also known as single-cell gel electrophoresis, is a widely used method for assessing DNA damage at the individual cell level. It was first introduced by Östling and Johanson in 1984 and has been popular in genotoxicity testing, environmental monitoring, and cancer research. This assay allows for the detection of various types of DNA damage, including single-strand breaks, double-strand breaks, alkali-labile sites, and oxidative base damage (Ostling, O., & Johanson, K. J.,1984).

The DNA in the gel was stained with the fluorescent dye acridine orange and gave a green emission in a microscope photometer. During the electrophoresis under alkaline conditions, DNA migrates towards the anode which is more pronounced in irradiated than in control cells. The intensity of the comet tail represents the number of DNA breaks.

Figure.1 Comet assay flow chart, figure derived from Breed Diagonsis.
Limitations: (Collins A. R., 2004)
  1. Lack of standardization:
    There is a lack of standardized protocols and guidelines for performing the Comet Assay. Azqueta, A., Gutzkow, K. B., Brunborg, G., & Collins, A. R (2011) and Ersson, C., & Möller, L. (2011) both concluded that the concentration of agarose, alkaline incubation time, particularly electrophoresis voltage and duration will hugely impact the result. These can lead to inconsistencies in results between different laboratories.
  2. Limited throughput:
    The Comet Assay is labor-intensive and time-consuming since there is a limit to samples processed in one experiment (Azqueta, A., & Collins, A. R., 2013). The manual nature of the assay makes it difficult to process a high volume of samples simultaneously, limiting its throughput and efficiency.
  3. Enzymes are required:
    Lesion-specific enzymes are essential, especially in human biomonitoring. Yet, the concentration of these enzymes must be high enough to detect all lesions present. Moreover, the enzymes are not entirely specific (Azqueta, A., & Collins, A. R., 2013).

As for our innovation, we will test the amount and level of DNA damage in vitro, which means people can obtain real-time data from living cells. In such a case, technicians can test the carcinogen in different time frames and dosages, and examine any chronic effects that the sample may have. By applying this novel method, scientists will only need to sustain a stable cell line transfected with the FRET pair and a confocal microscope to conduct this experiment. It will be more effective and less time-consuming, as neither they stabilize single cells on the plate with limited samples nor do they dye DNA with fluorescent dye. With the tested protocols provided on our notebook and protocols page, the execution of this analysis can be standardized and provide valid results.

Freedom to operate (FTO)

With support from our in-house experts and external advisors, we will employ both methods to report carcinogenic toxicity and DNA damage in our engineered cell systems. These methods include genetic activation-based RecA reporters and FRET-based systems.

To ensure that our projects are carried out without violating the intellectual property rights of others, we conduct an FTO (freedom to operate) analysis. A thorough search of patеnt databases such as Google Scholar, scientific literature, and other relevant sources was performed using keywords relevant to our project (e.g., "ATRIP FRET'' and "RecA DNA damagе"). We aim to identify any existing patents or intellectual property that may be relevant to our project and to analyze the identified patents and intellectual property to determine their relevance and potential impact on our project. Based on our research, we did not find any patent literature that is homologous to our research direction and concept, which is a positive outcome for our research project.

Outcomes and route to market

Market share gains can be influenced by various factors such as product differentiation, price, marketing strategies, and competition.

We assume that some popular industries such as the food industry will purchase to keep a check on the quality of their productions. Moreover, public health organizations and biotechnology companies can use our solution to screen the safety or toxicity of products on the market and devise new guidelines and measures to regulate them. We estimate that the system can be used to detect carcinogenicity of food and products in both industrial and public health settings. This helps determine whether food meets standards to protect consumers’ health and rights.

Market testing interviews play a crucial role in assessing the viability and alignment of a product with potential customers' needs. These interviews offer a unique opportunity to gather valuable feedback, understand customer preferences, and identify areas for improvement. Byengaging in open and insightful discussions with target companies, we can gain deeper insights into whether our product meets their specific requirements, allowing us to refine and tailor our offering to better serve the market's demands.

To effectively address critical aspects of preliminary market testing and entrepreneurship, such as budgeting, resource allocation, and profitability analysis, a primary estimation of operational costs has been undertaken. This estimation has been predicated upon the consideration of time, materials, and manpower required to complete a single cycle of our E. coli and HEK293 experiments.

However, in light of the inherent risks, facility constraints, and experiment complexities, several assumptions have been made to ensure a comprehensive analysis.

  1. It is important to note that all rent and utilities expenses have already been accounted for and covered by our school institute. As these expenses constitute basic costs without any additional charges, they have been excluded from the cost of goods sold and our current-stage manufacturing estimation for market testing.
  2. Our laboratory employs liquid nitrogen for the storage of human or animal cell lines. However, due to safety considerations, particularly for high school students even after training, we have opted to utilize a -80-degree freezer for the storage of all our product lines. Given that the current stock is primarily intended for market testing purposes, we will provide these products in tubes at no cost. Consequently, any limitations associated with the short-term storage of human cell lines in a -80-degree freezer will not have any adverse impact on the quality of the products.

By outlining these assumptions, we aim to ensure a comprehensive and accurate estimation of operational costs, taking into account the specific circumstances associated with our experiments. Please be noted that currency conversions and references were completed before 12th October 2023, with rate USD:HKD=1:7.82.


To ensure the production of high-quality carcinogen detectors, it is estimated that associated salaries will amount to up to $1359.64, which brings the total cost of production of stable transfected HEK-293 cell line and transformed E. coli stock to be $2645.2. This cost considers the involvement of skilled professionals and experts who play a crucial role in the production process. It also reflects the exceptional value, quality, and effectiveness of our product while remaining competitive within the market. We believe our product is a fair investment for organizations and individuals, taking into account the potential to save millions of lives through early detection. However, we due to uncertainties in production mode and clientele that cannot be confirmed at this stage, an accurate provision of a suggested market price cannot be suggested.

Due to the limitations stemming from the absence of advanced manufacturing equipment for our current market testing product, we have projected the production of 50 tubes of our cell assay reporter product, keeping in line with the previously mentioned scale and budget. Our strategy involves providing these samples free of charge to 20 target companies, which include Tigermed, WuXi AppTec, Crown Bioscience in mainland China, Celero, Parexel in Hongkong, and Novotech in Singapore, and subsequently arranging interviews to evaluate if our product aligns with their internal criteria.

Technical, commercial, and environmental risks

  1. Technical Risks:
    • Inability to Achieve Gеnеtic Activation: If we are unable to successfully engineer the E. coli strain through transformation to detect DNA damagе through the activation of the RecA-EGFP system, it may limit the efficacy of the enginееrеd E. coli strain in accurately detecting DNA damagе.
    • Mitigation: Wе will conduct thorough research, engage in careful design and optimization, pеrform itеrativе tеsting, and explore alternative methods until thе еxpеrimеnt is completed. Wе firmly bеliеvе that through our unwavеring efforts, we can minimize the risk to thе lowest possible lеvеl and achieve success in the thе engineering process of DNA damage detection.
  2. Commercial Risks:
    • Reduced Market Demand: In our previous work, a set of DNA-damage genes and others with specific functions have been discussed and analyzed with our in-house experts and external advisors, and we have chosen our target genes for developing our toxicity screening platform. However, developing a platform also requires a large amount of research and development funds, and higher costs may result in higher final prices for our products. Products with high pricing may face reduced market demand and limited market penetration as they may not be attractive enough to potential customers.
    • Mitigation: To address this high-cost issue, we have decided to еstablish basic production capacity by purchasing usеd machinеry. This approach allows us to reduce costs while also optimizing our еxisting rеsourcеs. Additionally, we hope to attract intеrеstеd invеstors and foster collaborations with morе businеssеs in thе future.
  3. Environmental Risks:
    • Sustainability and Environmental Impact: If E. coli or its biofilm were accidentally released from the laboratory into the environment, it could potentially have negative impacts. For example, it might lead to water contamination, disruption of natural ecosystems, transmission of pathogens to humans or animals, and the potential for the spread and persistence of the relevant strain.
    • Mitigation: To mitigate contamination concerns, we are engineering our E. coli to have significantly reduced or no biofilm. This allows us to remove bacteria based on less intensive UV light, ensuring the overall safety of the system while eliminating the need for incineration or chemical usage in decontamination.
Figure.2 Risks and Mitigation table

SWOT analysis

We decided to conduct a SWOT analysis as part of our strategic planning process to gain a comprehensive understanding of our internal strengths and weaknesses, as well as the external opportunities and threats in our operating environment. Ultimately, we were provided with valuable insights that will help us make informed decisions, allocate resources effectively, and achieve long-term success in an ever-changing business landscape.

Figure.3 Brief SWOT analysis of our proposed solutions

  1. Strengths: Our solution is largely independent of complex supply chains of reagents, which is not often the case for chemical or cell-based assays. With a stable supply of goods, we can maintain consistent production levels. This allows us to meet customer demand without fluctuations or disruptions that could result from supply shortages.
  2. Weaknesses: Due to the equipment and expertise needed to operate our solution, it may not reach widespread implementation and utilization. If our market is limited to university and hospital laboratories, we will not be able to make the desired impact on this industry.
  3. Opportunities: In the current market, despite existing similar products, other businesses may struggle to sustain their presence due to cost and material limitations. We can potentially establish dominance or even achieve market monopolization by investing in the training and popularization of our method.
  4. Threats: At present, the operation of cell-based assays requires a laboratory environment where safety is highly regulated. In order to market a relatively high-risk product, there is a need to meet the standards and regulations that surround this sector of products. When such laws a revised, we must work to meet the new requirements, which is both time-consuming and expensive.

Outline of Future Development

Considering our team members' immediate future plans, we have decided to proceed with the expansion of our startup in both Macau and London, UK. While Macau's company registration may take 1.5-2 months, the UK offers a swift registration process, achievable in just one day for £25 or £75-£125 with professional support from accounting or law firms. This dual-location strategy is designed to maximize our potential for securing funding and establishing valuable external collaborations. In Macau, we expect initial funding support from Wynn and other organizations, leveraging our strong existing relationship with Wynn. In the UK, we are actively preparing for the InnovateUK funding competition, ensuring our application aligns with the required format. With guidance from experienced tutors in the UK's biotech startup ecosystem, we are aiming for a score of 7/10, which, while not a guarantee of success, can provide access to support from InnovateUK EDGE for business startup training. This strategic approach also opens doors to various UK subsidies, such as the RTO/CATAPULT grant of £15,000 for further research and the UKIPO Audit and Assess grant of £7,500, which will shape our IP strategy and patent filings in future successes. Furthermore, our Macau establishment has the potential to extend its reach into Hong Kong, a path that many Macau entrepreneurs traditionally take. We've recently secured advisors with a proven track record in startups, who have received multiple rounds of funding from Hong Kong Science & Technology Parks (HKSTP). HKSTP provide a lot more support then the Macau government, including an impressive HKD 100,000 from the Ideation Programme, which empowers entrepreneurs to turn their brilliant ideas into reality and embark on their entrepreneurial journey. Additionally, we'll have access to a one-year startup support program tailored for tech-focused entrepreneurs. This opportunity for expansion and the strong backing of experienced advisors and funding sources further bolster our growth prospects; other subsequent funding scheme, such as Incubation and Incubio, provide HKD 1.29 to 6 million in 3-4 years.

Depending on the success, we are planning to establish a parent company in the final quarter of 2024, with a focused commitment to producing high-quality detectors until 2030. The market demands high-quality, reliable, and accurate detectors, and our goal is to earn customer trust and cultivate a strong reputation by consistently delivering accurate and reliable products. The increasing recognition of the importance of early cancer detection has created a growing demand for carcinogen detectors. With this opportunity in mind, our company aims to meet market needs and establish a significant presence.

As we solidify our brand image, our future "Next Half-Decade Plan" includes forming partnerships with local healthcare institutions, research organizations, and distribution channels to expand our product's market reach, creating more sales opportunities. By 2035, after a decade of continuous refinement and innovation, our profits will be reinvested to enhance product quality, streamline production processes, and introduce new features and functionalities. This ongoing innovation is essential for maintaining competitiveness and meeting the increasing market demand for advanced carcinogen detectors.

Our commitment to providing long-term quality support and after-sales services, including technical consultation, training, and maintenance, has always been a cornerstone of our company. These meticulous after-sales services are the foundation upon which we build customer loyalty and maintain a sterling reputation.

Figure.4 Outline of a possible direction of development for our project

References

Ferlay, J., Colombet, M., Soerjomataram, I., Parkin, D. M., Piñeros, M., Znaor, A., & Bray, F. (2021). Cancer statistics for the year 2020: An overview. International journal of cancer, 10.1002/ijc.33588. Advance online publication. https://doi.org/10.1002/ijc.33588

World Health Organization. (2022, February 3). Cancer. World Health Organization. https://www.who.int/news-room/fact-sheets/detail/cancer

National Institute of Health (NIH). (2021, October 11). What is cancer?. National Cancer Institute. https://www.cancer.gov/about-cancer/understanding/what-is-cancer

Centers for Disease Control and Prevention. (2017, July 6). Invasive cancer incidence, 2004–2013, and deaths, 2006–2015, in Nonmetropolitan and metropolitan counties - united states. Centers for Disease Control and Prevention.

Li, X., Deng, Y., Tang, W., Sun, Q., Chen, Y., Yang, C., Yan, B., Wang, Y., Wang, J., Wang, S., Yang, F., Ding, Y., Zhao, G., & Cao, G. (2018). Urban-Rural Disparity in Cancer Incidence, Mortality, and Survivals in Shanghai, China, During 2002 and 2015. Frontiers in oncology, 8, 579. https://doi.org/10.3389/fonc.2018.00579

Sharp, L., Donnelly, D., Hegarty, A., Carsin, A. E., Deady, S., McCluskey, N., Gavin, A., & Comber, H. (2014). Risk of several cancers is higher in urban areas after adjusting for socioeconomic status. Results from a two-country population-based study of 18 common cancers. Journal of urban health : bulletin of the New York Academy of Medicine, 91(3), 510–525. https://doi.org/10.1007/s11524-013-9846-3

European Commission. (n.d.). Carcinogenicity. EU Science Hub. https://joint-research-centre.ec.europa.eu/eu-reference-laboratory-alternatives-animal-testing-eurl-ecvam/alternative-methods-toxicity-testing/validated-test-methods-health-effects/carcinogenicity_en

Collins A. R. (2004). The comet assay for DNA damage and repair: principles, applications, and limitations. Molecular biotechnology, 26(3), 249–261. https://doi.org/10.1385/MB:26:3:249

Ostling, O., & Johanson, K. J. (1984). Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochemical and biophysical research communications, 123(1), 291–298. https://doi.org/10.1016/0006-291x(84)90411-x

Azqueta, A., & Collins, A. R. (2013). The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Archives of Toxicology, 87(6), 949–968. doi:10.1007/s00204-013-1070-0

Ersson, C., & Möller, L. (2011). The effects on DNA migration of altering parameters in the comet assay protocol such as agarose density, electrophoresis conditions and durations of the enzyme or the alkaline treatments. Mutagenesis, 26(6), 689–695. https://doi.org/10.1093/mutage/ger034

Azqueta, A., Gutzkow, K. B., Brunborg, G., & Collins, A. R. (2011). Towards a more reliable comet assay: optimising agarose concentration, unwinding time and electrophoresis conditions. Mutation research, 724(1-2), 41–45. https://doi.org/10.1016/j.mrgentox.2011.05.010