Team:Stockholm/Entrepreneurship
We, iGEM Stockholm 2017, believe that lung probiotics are the future, and we want to be at the forefront of this field together with PROlung.
To prepare PROlung to become a possible start-up in the future, we have conducted different parts of a theoretical product development strategy with the help of the book BIODESIGN The Process of Innovating Medical Technologies.
Pharmaceutical and biotechnological development are usually characterized by the development of a product looking for an uncertain market. In our opinion, the final customer should be always in the center of the development process and, because of this, we have used the product development strategy of the MedTech industry as our baseline, as we search for a way to provide a high-value solution for the patients.
In order to develop our product, having the final customer always in the center, we have included a section concerning this in the analysis of the disease state fundamentals, the existing solutions, the stakeholders involved in this area and the landscape of market for a treatment to respiratory diseases related to high mucus production. But most importantly, we have found the need behind our product, by listening to the opinions of different stakeholders.
Through the background analysis we performed and studies of customers’ needs, we were able to successfully develop the product design. For the product we had designed we made a theoretical study of the regulations it would be subjected to, the ethical concerns it may raise and the intellectual property protection we could apply for, and the product could interfere with. At the end of the process, we were also able to determine how the product could be reimbursed in healthcare and a possible business model for it. We believe that conducting all this would give PROlung a head start in becoming a company.
Since we know the target audience for this document has a very varied background, we have included, in the beginning of each segment, a short description of its purpose. We hope that this description of and business concepts and the word list of complex medical terms we have also included can inspire future teams to think about the product development of their projects.
We have divided our product development in two different parts: the journey to finding a need and the journey to putting a product in the market. We have taken this decision for a matter of simplicity and thought this is the key point to do so as the product design step of the product development is a turning point for the innovator in which there is a change from a more social, value based mindset to a more utilitary and practical one.
Disease state fundamentals is establishing a detailed knowledge of the relevant disease state, with a particular focus on its mechanism of action. A disease state fundamentals is key to validating any need and understanding how it can best be addressed.
Physiology:The way in which bodily part functions
The human lungs are a pair of large, spongy organs optimized for gas exchange between our blood and the air, as our bodies require oxygen in order to survive.
The air that you breathe goes down your windpipe into tubes in your lungs called bronchial tubes, that branch many times into smaller, thinner tubes called bronchioles. These tubes end in bunches of tiny round air sacs called alveoli. Small blood vessels called capillaries run along the walls of the air sacks. To protect the body from these foreign particles, the lungs are covered in something called mucus1.
When air reaches the air sacs, oxygen passes through the air sac walls into the capillary blood At the same time, a waste product, carbon dioxide (CO2), is removed from the blood, into the air sacs. This gas exchange process brings in oxygen for the body to use for vital functions and removes the CO2.
The airways and air sacs are elastic breathing in, each air sack fills up with air, like a balloon, when breathing out, the air sacs deflate and the air goes out.
The mucus that covers the lungs is a viscous substance which traps these foreign molecules and consequently, prevents them from entering the body. Tiny hairs called cilia constantly move the mucus with the entrapped pathogens upwards and out of the lungs. Thus, the mucus functions as a protection against the outside world, continuously keeping the airways clear of foreign particles.2
Analysing the existing solutions is to explain the current and emerging solutions, how they work, when they are used, how effective they are, their cost and the overall value they deliver. The aim is to clarify the gap in existing solutions, where new opportunities become apparent.
Bullectomy: Surgical removal of a bulla, which is a dilated airspace in the lung parenchyma
The most common medications for cystic fibrosis, COPD and asthma are the inhalation devices like bronchodilators, mucus thinners, and steroids. A famous new drug on the market is Orkambi. Thick mucus also contributes to bad bacteria in the lungs were often antibiotics are necessary. These medications are always taken together with airway clearance techniques and in severe cases, a lung transplant may be needed.
Current commercially available approved drug class are:
Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) modulators (ivacaftor (Kalydeco®) and lumacaftor/ivacaftor (Orkambi®).) The Lumacaftor moves the defective CFTR protein to the correct place on the cell surface, and ivacaftor increases the protein's activity once it is in place. Orkambi is today one of the most attractive medications for CF but the therapeutic effect only targets a subpopulation people with specific CF mutations 3.
Bronchodilators is an inhalation device that helps widen the airways, and can also be used for asthma and COPD patients. There are two different types, short-acting and long-acting and can be used before other treatments and before exercise.
Mucus thinners, such as Mucolytic drugs is a type of inhalation medicine that alters the chemical characteristics of mucus to decrease its viscosity and facilitate its removal. There are two types hypertonic saline works by increasing the amount of sodium in the airways and Dornase alfa (a DNA functions by cutting DNA strands released by white blood cells, which makes the mucus thinner.
Potential disadvantages of inhalation medication in CF patients compared to other drug administrations are:
Uncertainty about drug dose at the target site, severely affected lung areas may not be reached, drug delivery depends on inhalation technique and device performance, local side effects (e.g., cough, airway narrowing, hoarseness), variable systemic drug absorption, time-consuming drug administration, need for education and training, limited information on drug interactions in the lung, specific drugs may need specific delivery devices, poor adherence, potential pollution of the environment, potential device contamination and patient infection, need for hygiene control and maintenance of the equipment and limits social functioning4.
Anti-infective drugs include antibiotics and antibacterials, antifungals, antivirals, and antiprotozoals. These work by either killing the infectious agent or by inhibiting it from spreading.
Antiinflammatory drugs have two main purposes, to act as painkillers or to reduce inflammation5.
Pancreatic enzyme supplements are taken by mouth with the purpose to digest carbohydrates, proteins, and fats, absorb essential nutrients such as vitamins and minerals and help the patient gain and maintain a healthy weight6.
Lung Transplantation can extend and improve patient’s quality of life, but it involves an extensive evaluation process and a commitment to living the lifestyle required to keep your new lungs healthy.
Bronchodilators (mentioned above).
Inhaled steroids such as inhaled corticosteroid medications can reduce airway inflammation and help prevent exacerbations. Side effects may include bruising, oral infections and hoarseness.
Oral steroids work for patients who have a moderate or severe acute exacerbation. Short courses (for example, five days) of oral corticosteroids can prevent further worsening of COPD, however, long-term use of these medications can have serious side effects, such as weight gain, diabetes, osteoporosis, cataracts and an increased risk of infection.
Phosphodiesterase-4 inhibitors is a new type of medication approved for patients with severe COPD and symptoms of chronic bronchitis. One example is roflumilast (Daliresp), a phosphodiesterase-4 inhibitor. This drug decreases airway inflammation and relaxes the airways. Common side effects include diarrhea and weight loss.
Theophylline is a medication, which might help improve breathing and prevent exacerbations. Side effects may include nausea, headache, fast heartbeat and tremor. Side effects are dose-related, and low doses are recommended.
Antibiotics can act against respiratory infections, such as acute bronchitis and pneumonia.
Surgery is an option for some people with some forms of severe emphysema who aren't helped sufficiently by medications alone. Surgical options include:
Asthma medications are divided into two main parts, long term, and short term medication, where the first acts to reduce airway inflammation, which helps prevent symptoms from starting and the second one acts to give rapid, short-term symptom relief during an asthma attack. Asthma has very similar medications as COPD such as the inhaled steroids, bronchodilators and Theophylline.
Other long-term control medicines include:
Short terms include:
In addition to medication, techniques can be taught by a physiotherapist to help restore clear airways in all of our targeted diseases. Such exercises includes: active cycle of breathing techniques (ACBT), a technique that involves a sequence of relaxed breathing, followed by deep breathing exercises and then huffing, autogenic drainage, a series of gentle breathing techniques that clear mucus from the lungs, modified postural drainage, a technique that involves changing your position to make it easier to remove mucus from your lungs and airway clearance devices, a handheld devices that use vibration and air pressure to help remove mucus from your airways9.
What is this?
There are many different stakeholders with a potential interest in new medical technology. Those include patients, patient’s families, patient advocacy groups, physicians, professional societies, nurses, family administrators, public payers, governments and non-governmental organizations.
Some examples of different societies and and non-governmental organizations in Sweden are Riksförbundet Cystisk Fibros, Astma- och Allergiförbundet and Riksförbundet HjärtLung. The majority of the countries with people suffering from these diseases have similar associations for these diseases10.
n Sweden, the responsibility for medical care and health is shared by the central government, county councils, and municipalities, and therefore they are the decision makers. They are also the payers as the major population in Sweden possesses public insurance. The county council is responsible for ensuring that the needs of tomorrow are met by today, in healthcare.
The doctors and government officials influence the upcoming treatments on the market, and the user is, of course, the patient.11
What is this?
To identify the need, we targeted our most valuable stakeholders, the patients and healthcare professionals. We conducted a survey to reach out to as many patients as possible with the results that there is a clear need for a medication treating mucus, they are happy to try GMO as a drug and the best device to use would be a nebulizer. We arranged interviews with doctors, physiotherapists, nurses and family members, with a similar outcome as with the patients. There is a need for PROlung and nebulizer would be the most effective device. They all would be happy to prescribe PROlung to their patients as long as it’s safe.
To find the need for a new medication, we decided to focus on our most valuable stakeholder, the patient. We did this is numerous ways. We found that the easiest way to gather as many opinions as possible is through a survey. We made it accessible in both English and Swedish in order to target as many patients as possible. We reached out to the Facebook group “Cystic Fibrosis” with almost 30 000 members, disease organizations such as “Riksförbundet Cystisk Fibros”, ”Astma & Allergiförbundet” and “ Riksförbundet HjärtLung” who posted the survey in their newsletters and social media pages. We also attended the biggest cystic fibrosis event in Sweden, where we presented our project and talked to patients about the need for a new medication.
From the patient's surveys, we can very easily interpret the results that there is a need for medication targeting mucus. Overall, 228 asthma patients, 41 CF patients, and 10 COPD patients answered our survey. The reason that a majority of the answers came from asthma patients may be that asthma is a much more common disease than the CF and COPD and we needed to account for this as we analyzed our results as the diseases have slightly different needs.
The questions asked to patients:
Up to 50 % have changed their treatments more than 5 times, which stated that the current medications don’t work sufficiently enough for longer time use. 20% are very satisfied with their current treatments, 45% are satisfied and 35% are not satisfied.
About 50 % answered they have issues with excessive mucus. In this answer, we also have to take into account that 228 asthma patients answered, which don’t suffer from mucus in the same amount as COPD and CF does. The majority that answered the survey does not think that their current treatment is good enough against the mucus. This is probably our most interesting answer as this clearly states that there is a need for a new effective medicine targeting mucus. Although we have to take into account that over 60 % are satisfied with their treatments. We drew a conclusion by going through surveys individually, that the patient's suffering the worst from mucus, are not satisfied with their current treatment as it does not help the excessive mucus, whilst those that have a milder mucus overproduction, are more satisfied with their current treatment. About 40% said yes to a medication being a GMO, and the rest stated maybe. 50% said the most suitable device would be any type of inhaler. The rest of the solutions, such as an orally dissolving tablet (ODT), injection and a surgically implanted device had a similar small percentage.
From the results we obtained from our surveys, we decided to create personas. Personas are fictional characters, which you create based upon your research in order to represent the different user types that might use your service, product, site, or brand in a similar way. By writing about personas we are able to emphasize the need and to put our reader in the skin of the person suffering the disease.
Additionally, we decided to target the second largest stakeholder, the doctors, physiotherapists, and nurses. We knew we couldn't target an equally big group as with the patients, and therefore decided to conduct interviews with six professionals in healthcare.
We interviewed Michael Runold, a pulmonary physician, treating all kinds of pulmonary diseases mainly COPD; Niclas Johansson, a physician in infectious diseases, treating mostly respiratory infections like tropical fever and COPD; Annika Hollsing, a physician in pulmonology and infectious diseases, treating mostly CF and asthma; Robert Dickson, a pulmonary disease and internal medicine doctor working with pulmonary diseases, specifically COPD and asthma; Pernilla Sönnerfors, a pulmonary physiotherapist for mostly asthma and COPD; and Ulrica Sterky, a children’s nurse and also the mother to a son with cystic fibrosis.
Questions asked to healthcare professionals:
The symptoms were very similar for all answers, such as cough, shortness of breath, tightness of the chest, breathing problems and mucus. The same goes for the prescribed medications, including for an example steroids, bronchodilators or antibiotics. The mucus targeting medications that were mentioned were DNAse, acetylcysteine, bromhexine, sodium chloride, mucolytics and extensive training on how to eliminate the produced mucus, but they all agreed that there is a gap in medications targeting COPD, asthma, and CF and they insisted strongly that there is a need for a medication specifically targeting mucus, especially as the ones today do not work for all patients. This was also encouraged by Pernilla, the physiotherapist that said:
"There is absolutely a need, it would be a great win for those that have a hard time moving the thick mucus upwards. If there was a good enough mucus targeting solution today, the patients would not have to visit me, there is definitely a need."
We asked Ulrica, children's nurse and mother to a son with cystic fibrosis “What do you think it is like living with cystic fibrosis?”
Her answer:
“In the big picture, when your child gets the diagnosis, it's a very hard time for patient and families. You have to keep track of the infections, make sure they eat enough with their enzymes, practice the breathing exercises and make sure they do them correctly and always in the back of your head, you know it's for life. You just have to learn to live with it, knowing that many don't survive over middle age years.”
All the participants addressed GMO’s as a medication very positively. They said as long as it’s safe and approved, and of course works, why not?
Additionally, when we asked what device would be the most suitable, the questions were quite similar but with some differences. To summarize, the patients are quite tired of using inhalers and the ones today are quite complicated. If the medication could be given as a pill, that would be the easiest. Although considering side effects and the fact that the pill might not deliver the medication as specific as wished, all of them agreed that an inhaler like a nebulizer would be the best. More specifically, it would be the easiest for the patients if the inhaler was small enough to fit into a pocket.
In regards to the possible market of our potential solution, about 40% of the surveyed patients said yes to a medication being a GMO, and the rest stated that maybe they would buy it, giving our product a very interesting number of potential customers.
On the physician side, all the participants addressed GMO’s as a medication very positively, said that as long as the product is safe and effective, they would not have any problem prescribing it.
Overall, we have been able to understand what the priorities the health care professionals and patients have for the use of PROlung. Additionally, from the surveys and interviews, we were able to identify the different customers, who have stated interest in this solution and would be willing to buy it.
What is this?
We aimed to map the how well customer needs are met, key players and stakeholders and market size, is it expanding or contracting? As discussed, the need is not fully met as current mucus treatments aren't efficient enough. Key players are companies as Vertex, AstraZeneca, GlaxoSmithKline, F. Hoffmann-La Roche, Novartis, Merck and more. Key stakeholders are doctors and patients. The market for cystic fibrosis is growing due to increasing prevalence with a size of $ 3.56 billion. For COPD and asthma, the market is expanding, with asthma having the largest market size of $ 20.7 billion and COPD, $ 11.3 billion. North America is the largest market for all three diseases but Asia Pacific is expected to become the fastest regional market in the future.
Prevalence: The amount of a population who have/had a specific characteristic in a given time period
Doing the market analysis, we took help from the book Biodesign, The Process of Innovating Medical Technologies by addressing following questions (in italic) to analyze the market of our target diseases.
We were able to clarify these questions by addressing the market with a quantitative and qualitative research, by both looking at existing data and also doing surveys and interviews with current stakeholders.
How well are customer needs generally addressed by existing solutions and how closely aligned are available solutions with the need the innovators are seeking to address?
There are no solutions today that target the specific area of breaking down the mucus directly. As read under existing solutions for cystic fibrosis, there are so-called mucus thinners, but they do not degrade the mucus directly and serve more to make it thinner by adding salt or using DNase.
Especially in cystic fibrosis and COPD, mucus is a major problem, which current treatments do not satisfy completely. For asthma, on the other hand, mucus is more of an indirect issue, and tightness of airways is the major problem, breaking down mucus could act as a complement to airway dilators.
What is the approximate size of the total market for all existing solutions? Is the market expanding or contracting?
Cystic Fibrosis:
The increasing prevalence of cystic fibrosis (CF) (In the U.S., the number of patients in 2010 rose from 26,366 to a total of 28,983 patients in 2015)) together with rising treatment rate is one of the key reasons for the market growth. In upcoming years, an increase in R&D funding by both private and public organizations, a rising number of initiatives undertaken by nonprofit organizations, and presence of favorable reimbursement policies are expected to drive the market.
COPD:
The global market size of COPD is expanding. Much of this growth is a result of a high number of new, more efficacious and convenient products entering the market and commanding greater value compared to the therapies already in the market.
Asthma:
Although asthma has the biggest market size, it is anticipated to show a slow growth rate of 2.2% over the forecast period from 2016 to 2024. And increasing incidence of respiratory diseases together with the demand for cost-effective treatment options is expected to push the asthma therapeutics market growth during the forecast period. According to the estimates published by the Global Initiative for Asthma (GINA), in the year 2013, around 300 million patients globally were reported to be afflicted with this disease and the number is expected to grow in the coming years11.
What is the size of the market opportunity in each market segment and the potential for growth and expansion?
Cystic fibrosis:
The market for cystic fibrosis is segmented into pancreatic enzyme supplements, mucolytics, bronchodilators, and CFTR modulators.
In 2016, CFTR modulators accounted for the largest revenue with a share of approximately 47.3% owing to the launch of new drug Orkambi. This drug is responsible for correcting the function of a defective protein caused by the CF gene.
The market is also segmented into oral route and inhaled drugs. Oral route drugs account for the largest share of nearly 63.7%.
According to the U.S. patient registry 2015, hypertonic saline was prescribed to 21.5% of children in the age group of 0 to 3 years and 41.9% children in the age group of 3 to 5 years. Increasing prevalence of CF among newborn babies is one of the key factors responsible for growing demand for oral drug formulation.
Country wise, the market is segmented into North America, Europe, Asia Pacific, Latin America, and Middle East and Africa. North America accounted for the largest revenue of $ 2,396.4 million in 2016. CF is most prevalent in North America.
Asia Pacific is expected to be the fastest growing segment with the lucrative growth rate of 18.8%.
Currently according to the Cystic Fibrosis (CF) Therapeutics Market Analysis By Drug Class, the availability of CF-specific drugs is low, however, growing approvals and product launches in this market are anticipated to impel growth.
COPD and Asthma:
Key drugs employed for control and diagnosis of asthma and COPD may be classified on the basis of their types which include bronchodilators, leukotriene antagonists, and immunosuppressants.
For asthma, the combination therapy segment including inhaled corticosteroids with long-acting beta-agonists dominated the overall therapeutic class market in 2015. This segment captured over 48.0% of the global market share and this drug class boasts of the following benefits: enhanced therapeutic effect, extensive availability, and enhanced patient medication adherence.
The inhaler segment for asthma includes dry powder, metered dose, and soft mist inhalers. The metered dose inhalers (MDIs) segment dominated the overall market in 2015 in terms of revenue share at over 45.0%. This is majorly attributed to, among others, reliability, versatility, self-containment, easy availability, portability, and cost-efficient medical aerosol delivery option. Increasing requirement for emergency treatment options aimed against sudden asthma attacks and portable devices constitute the crucial determinants contributing towards the largest market share possessed by the dry powder inhalers (DPIs) product segment.
Nebulizers demonstrate significant benefits in the home healthcare section which is expected to provide this segment with future growth opportunities. Nebulizers are small, portable, and able to dispense medication directly into the respiratory tract. These associated benefits are expected to boost its usage rates in the future13.
The growing aging population coupled with increasing automotive and industrial exhaust gases have remained the key drivers for the global asthma and COPD market, especially in the low-income population in the emerging economies of China, India, Brazil, and Russia.
Increasing disposable income levels in emerging economies as well as developing countries of Latin America, Middle East, and Africa has led to the increased usage of asthma pumps and sprays in these regions. However, the unknown etiology of asthma and COPD in medical industry may restrict the market growth in the next six years. Upcoming treatment methodologies such as once-a-day combination products and monoclonal antibodies have created new opportunities for market growth.
New bio-based medications in combination with biotechnology promise immense growth opportunities along with sustainable development for asthma and COPD market over the next decade.
Currently, North America leads the global market for asthma and COPD drugs and devices, followed by Europe in terms of market capitalization. However, because of the expiration of several patents, these regional markets will lose out some of the market shares to other emerging regional markets. Asia Pacific is expected to become the fastest regional market for asthma and COPD14.
Who are the key players of the market?
For cystic fibrosis, the key companies are AbbVie, Inc.; F. Hoffmann-La Roche Ltd.; Gilead; Novartis AG; Vertex Pharmaceuticals Inc.; AIT (Advanced Inhalation Therapies); Alaxia; Teva Pharmaceutical Industries Ltd.; Merck & Co., Inc.; Alcresta Therapeutics, Inc.; Allergan; and AstraZeneca15.
For asthma and COPD, the key companies are AstraZeneca, GlaxoSmithKline, F. Hoffmann-La Roche, Novartis, and Merck. Other significant companies operating in the global asthma and COPD market include Abbott, Amgen, Actavis, AptarGroup, Aurobindo, Boehringer Ingelheim, Cipla, Dr. Reddy’s Laboratories, Glenmark, Mylan, Pfizer, Ranbaxy, Sunovion, and Vectura.
Which stakeholder could recognize the greatest value created by a new solution?
The patient is the most valuable stakeholder for all three diseases. Their life quality is affected by their sickness, and they are the ones carrying the daily difficulties to get rid of the mucus, Finding a new medication that works affects them the most and, they are also the ones ready to take a “risk” developing GMOs, for a better life.
What is it?
The solution iGEM Stockholm has developed for the need “a treatment for diseases with excessive mucus production” is a smart genetically modified bacteria that secretes mucus degrading enzymes. The solution we have developed combines a nebulizer for the administration of the genetically modified bacteria, the genetically modified bacteria and the enzyme that will degrade the mucus in the lungs.
A nebulizer is a device that converts a liquid into aerosol droplets suitable for patient inhalation using pressurized air. A nebulizer is comprised by, a compressor, a container holding the medication to be administered, a tube connecting both parts and a mouthpiece or mask for the administration of the medication.
Particle size is of great importance at the site of deposition in the respiratory tract of humans and animals. A Collison nebulizer has been previously used to generate small-particle aerosols with a mass median aerodynamic diameter (MMAD) of 1 to 3 μm for animal exposure studies, this correlates with the small particles around the size of a bacterium (1 to 5 μm) that would deposit in the alveoli.16
The advantage of using a nebulizer instead of a pocket inhaler is its normal breathing medicine intake rate. By this, we mean that for the inhalation of medicine through a pocket inhaler the patient needs to take deeps breaths for the medicine to reach the lungs, however, for patients with a respiratory disease, this may not be a possibility due to the narrowing of the airways during these diseases. Additionally, nebulizers can deliver short-acting or long-acting medication therapies .
For the bacteria to survive, the administered medication placed in the nebulizer will need to contain the GMO plus a medium which is required for the viability of the bacteria.The most probable scenario would be that bacteria will be in vial protected and preserved and upon administration, when it will be inserted into the nebulizer and inhaled. It should be noted that it is wise to avoid keep GMO in the nebulizer container while there is no use in order to decrease the risk of release into the environment.
The advantage of using a nebulizer instead of a pocket inhaler is its normal breathing medicine intake rate. By this, we mean that for the inhalation of medicine through a pocket inhaler the patient needs to take deeps breaths for the medicine to reach the lungs, however, for patients with a respiratory disease, this may not be a possibility due to the narrowing of the airways during these diseases. Additionally, nebulizers can deliver short-acting or long-acting medication therapies.18
There could be other alternatives for the administration of the genetically modified bacteria, like microaspiration and dispersion of oral bacteria into the lungs. However, diseased lungs have the tendency to create niches for pathogens and neutral distribution does not affect the deposition of bacteria. Also, it would be even more difficult to assess the amount of bacteria practically reaching the lungs. So far, no models have been successful describing this neutral distribution. At last, it would be optimal if the administration minimizes the risk of any release into the environment which does not seem optimal upon oral or nasal application.
Once the bacteria reach the lungs, it would be necessary to control the dynamic process where selective pressure exists while mucociliary clearance and immune system strive to eliminate any foreign particle, for the bacteria to be able to stay long enough to release the enzyme.
Our proof of concept has been generated in E.coli, since it is a well-characterized bacteria suitable for engineering. It became clear to us that the next step would be to transfer this genetic construct to a strain more suitable for a lung probiotic.
In order to achieve this goal we considered two main approaches. On one hand to extrapolate our system from E.coli to a new bacteria we proposed the use of shuttle vectors, plasmids compatible with two different species which allow us to move cellular machinery from one bacterial chassis to another.
Our basic selection criteria was therefore based on two premises: the selected bacteria should have a shuttle vector compatible with E.coli and it should be one of the species that are part of the lung microbiota.
After reviewing the available shuttle vectors on the iGEM Registry of Standard Biological Parts we came down to two strong candidates: Pseudomonas aeruginosa and Staphylococcus aureus.
Although both organisms are well defined within the lung microbiota and have available shuttle vectors, the higher compatibility of P. aeruginosa for genetic manipulation would make it a better choice. Nonetheless there is one major drawback for the use of either of these organisms: its pathogenicity. Our initial strategy to cope with this limitation would be to begin with a lung-adapted isolate of P. aeruginosa that doesn’t produce virulence factors.
Another option within this approach could be the use of the part BBa_J153000. This shuttle vector was originally designed to work on cianobacteria that holds the potential to work with a broad range of gram negative . This would allow us to attempt to use non-pathogenic gram negative strains from predominant genus such as Fusobacteria or Veillonella.20
On the other hand taking into account the constant advancements in genetic engineering we could consider another approach with a lesser focus on shuttle availability. A great example of this would be the use of S.carnosus, a non-pathogenic strain for which a shuttle vector is currently under development and has successfully expressed hyaluronate lyase from E.coli.21
Our hypothesis is that our lung probiotic will reside naturally in the lungs of the patients (bronchi, bronchioles); on the mucus layer. As the osmotic pressure of the mucus is higher in the previously mentioned diseases we took advantage of this, by introducing an osmotic promoter in our bacteria and place the gene of interest, in our case, an enzyme called sialidase, under its regulation. Thus, when an abnormal osmotic pressure is detected by the bacteria, the gene expression of the mucus degrading enzyme would subsequently be triggered.
Once the enzyme is expressed and secreted, sialidase will cut the sialic acid in the branches of the thick mucus. Altering the mucus composition would make the mucus thinner and restore the mucociliary clearance.
We have mainly focused on two enzymes. Firstly sialidase, an enzyme that hydrolyzes glycosidic linkages of terminal sialic acid residues in glycoproteins. These sialic acids have been suggested to inhibit mucin degradation. Using a genetically engineered bacteria to remove these protective terminal groups will improve interaction and degradation of the mucins. Secondly, endo-β-galactosidase, is an enzyme that hydrolyzes the bonds next to galactose saccharides in the polysaccharide chains of mucins. It has been proven to release saccharide chains from glycans expressed in the gastric mucous cell-type mucin.22
In the mucin constructs, saccharides interact with water and form a mucosal layer. The large mucin constructs give the mucosal layer its visco-elastic properties. By using the aforementioned enzymes, these polysaccharides will be removed from the protein backbone, resulting in the mucus losing some of its gelatinous attributes.
There are two main differences between using an enzymatic approach instead of using mucolytics. The first one is the specificity. Enzymes are highly specific in comparison to other chemicals and will only interact with the mucin as well as often have a reaction time that is several times faster.The second difference is that the bacterial approach provides an autonomous system. This system will regulate the expression and secretion of the drug/enzyme accordingly to the disease state. More specifically, it is not until the bacteria can sense the elevated osmotic pressure that it will subsequently initiate the expression of the enzyme(s). The conventional therapy of mucolytic simply attempts to provide a one-time bolus to restore the mucus state, limiting its effects to being temporary.23
The way our system is constructed, the amount of enzyme expressed and secreted in the lungs will be associated and proportionate towards the osmotic pressure, making our treatment a personalized medicine. In contrast, a patient using mucolytic will have to try to decide on his/her own what dosage is needed for that particular time.24
When the mucus has been degraded, and thus its visco-elastic properties have been restored, the cilia are once again able to move the mucus, and hence restore the mucociliary clearance. The superfluous mucus can subsequently be coughed up or swallowed.
Product design was one of the most complex parts of our product development, and one of the ones where we required more help. For the successful completion of this part, we had support from Andreas Lundquist. Andreas holds two masters, one Technical Design and one in Mechatronics. He was a Clinical Innovation Fellow for the Center for Technology in Medicine and Health, and is currently the course leader for the master level course Product Development in the Biomedical Industry at the Karolinska Institutet.
What is this?
Patent: Exclusive right to sell, license or use an invention, a product or a process of the human intellect
We have developed a patent strategy for our product while fulfilling the biobrick sharing agreement. In order to do so, we have divided our patent search into three parts, lung therapeutics, E. coli GMO and drug enzyme. After executing a patent search, we have analyzed all the patents that are related to our project and we were not able to find any patents combining the three parts of our search. The next strategic step, if we wanted to continue with the development of our product and create a company out of it, would be to file a provisional patent application before we present our results in the Jamboree.
One of iGEMs values is sharing, an example of this is the biobrick agreement, where you share your biobricks and therefore get access to other biobricks. A problem arises for those that wish to continue with their iGEM project as a start-up, as everything that is shared can not be patented. Many of the iGEM teams that want to start a business, could do so by patenting program codes instead of sequences or similar25 26.
Still, one of iGEM’s greatest achievements is the number of startups it has generated. So, is there a way to keep sharing, but still be able to run a start up after iGEM?
We in iGEM Stockholm have - with the help of IP experts - completed a strategy to turn an iGEM project into a startup, while still respecting and acknowledging iGEM values. This will be exemplified with our project, PROlung.
In terms of IP strategy, our project can be divided into three parts:
In an initial patent search, we would firstly, have to look for patents inside the field of A, B or C, and see if any of those are relevant for our research. Secondly, we would need to study the combinations of A and B, B and C and A and C, in order to obtain a deeper understanding of the current situation in this field. Thirdly, we would have to see if there is any patent as a combination of A, B and C, that could have similar, or the same claims of our intended patent. All of this in addition to all scientific papers published before the filing date of the patent counts as prior art to our invention and could invalidate its patentability. If our invention is able to have a characteristic feature that does not infringe any of the explained options and combinations, it would be patentable.
With the help from Rosa Lönneborg, a lecturer in patent search and critical information literacy inside the life sciences at the KTH Royal Institute of Technology, we were able to develop our patent strategy and to identify patents related to our product.
Using the strategy previously mentioned, we were able to perform a patent search describes how current and expired patents relate to the product we had developed:
Patents covering Part A:
Product Development
Our Approach
Disease state fundamentals
What is this?
Word list
Epidemiology:The study and analysis of the patterns, causes, and effects of health and disease conditions in defined populations
Pneumothorax: There is a tear in the lung and air escapes into the chest cavity
Dyspnea: Shortness of breath
Recurrent sinusitis: The small, air-filled cavities behind your cheekbones and forehead get inflamed
Mucociliary clearance: Self-clearing mechanism of the bronchi
Anatomy of the lungs
Physiology of the lungs
Existing solutions
What is this?
Word list
Osteoporosis: Bones becoming less dense due to the loss of bone material
Exacerbation: Increase in the severity of a disease
Hoarseness: Abnormal voice changes
Summary
Cystic Fibrosis
COPD
Asthma
Airway clearance techniques
Stakeholders
Stakeholders are the parties that are directly and indirectly involved in financing and delivering care for the patient. In this analysis, you uncover who the user, payer, decision maker and influencer is.
Need
Needs correspond to opportunities for innovation. They are characterized by describing an outcome that is currently unmet for a problem, in a particular population, which helps direct the opportunity. By clearly and concisely articulating the needs they have observed, innovators will be in a much better position to then determine which ones represent the most compelling opportunities.
Summary
How long have you been diagnosed with this disease?
Approximately, how many times do you visit the doctor every year?
How many hours a week do you perform any type of physical activity?
Have you been trained on exercises that can improve the capacity of your lungs and help in the progress of the disease?
What is your current treatment? If you are not sure of the name, please describe your medication or state unknown.
How many times have you changed treatment?
Are you satisfied with your current treatment?
Do you have problems with excessive mucus?
On a scale of 1-5, how much has your current treatment improved your excessive mucus? (1 being not improved, 5 being highly improved).
If a new treatment was to be released, outstanding in the performance of the current ones, would you take it if it involved the use of GMOs (Genetically Modified Organisms) in the body?
If this treatment was to be released, which type of device would be more suitable for its administration?
David
David is 12 years old and was diagnosed with Cystic Fibrosis when he was just a baby. David loves being outdoors and doing sports, but unfortunately, he has to spend many hours inside taking his medicines and doing his breathing exercises. Last week he missed three classes in school due to his doctor's appointments, and now he has to change medicine again, as the current one is not as optimal as before. David hopes that he will grow older than his grandparents and that in the future there will be a cure for cystic fibrosis.
Anna
Anna is 56 years old and suffers from COPD. Anna loves spending time with her grandchildren but feels ashamed to be out in public with them as her coughing draws too much attention. Anna doesn’t know how many more years she has to live and all she wants is to be able to see her grandchildren grow up. Her lung function is very low and she hopes for a lung transplant, although many people stand in line before her. At this point, Anna is open for any type of treatment.
What is your background?
What are the symptoms you check for when you have an appointment with a patient?
What type of medications do you prescribe/prefer to the different diseases/what exercises do you teach them?
Is there a gap in medication for cystic fibrosis, asthma, and COPD?
How do you treat patients who have problems with mucus?
Would there be a need for a medication targeting mucus?
Would you be open to prescribing a drug involving the use of a GMO?
If this GMO treatment was to be released, which type of device would be the most suitable?
Market
Market analysis aims to give a broad understanding of the need in terms of its total size, range of existing solutions and competitors, and gaps that may exist and indicate opportunities for innovation. It also focuses on a detailed evaluation of different segments within the overall market to determine which one(s) the innovators should potentially target.
Summary
Word list
R&D: Research & Development
Reimbursement: To pay back money to someone who has spent it for you or lost it because of you
Value: A measure of the benefit provided by a good or service to an economic agent
Revenue share: The distribution of profits and losses between stakeholders
Market capitalization: The total market value of all of a company's outstanding shares
The Global Market Size
$3.56 billion
The Global Market Size
$11.3 billion
The Global Market Size
$20.7 billion
Product Design
Based only on information about the need, innovators are able to develop promising solution concepts, trying to create something technically feasible, viable, efficient… The final concept will continue to be refined, tested, iterated, and improved during development and implementation.
Summary
Word list
Microaspiration: Used when patients have an asymptomatic aspiration of small volumes of oropharyngeal secretions or gastric fluid into their lungs.
Vector: DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell.
Microbiota: Microbes colonizing the human body, includes bacteria, archaea, protists, fungi and viruses
Glycosidic linkages: Covalent chemical bond joining a carbohydrate molecule to the group to which it is attached (which may be in another sugar, as in polysaccharides, or in non-sugar molecules)
The nebulizer
The genetically modified bacteria
The mucus degrading enzyme
Intellectual Property
Intellectual property (IP) refers to inventions such as literary and artistic works, designs and symbols, names and images used in the industry and is protected by law with patents, trademarks, and copyrights to gain financial benefit or recognition from what you’ve created or invented. Source.
Word list
Provisional patent: Allows the inventor to file for the priority date of an invention without a formal patent claim, oath or declaration, it has a one year validity, after which a patent application should be filed or dropped.
Summary
Result
Result:
With the help from Rosa Lönneborg, a lecturer in patent search and critical information literacy inside the life sciences at the KTH Royal Institute of Technology, we were able to develop our patent strategy and to identify patents related to our product.
Sonia and Amanda working on patents together with Rosa Lönneborg
Using the strategy previously mentioned, we were able to perform a patent search describes how current and expired patents relate to the product we had developed:
Patents covering Part A:
US20150017150A1- METHODS, COMPOUNDS AND COMPOSITIONS FOR TREATMENT AND PROPHYLAXIS IN THE RESPIRATORY TRACT27
WO2015184526A1 SOLUBLE BACTERIAL AND FUNGAL PROTEINS AND METHODS AND USES THEREOF IN INHIBITING AND DISPERSING BIOFILM28
WO2008001045 A1 MEMBERS OF THE GLYCOSIDE HYDROLASE FAMILY 31 FAMILY OF PROTEINS29
WO03010298A1 PROBIOTIC LACTOBACILLUS SALIVARIUS STRAINS30
WO2003010299A1 PROBIOTIC LACTOBACILLUS CASEI STRAINS31
Patents covering Part B:
WO2003010297A1- PROBIOTIC BIFIDOBACTERIUM STRAINS32
Patents covering Part C:
US20150017150A1- METHODS, COMPOUNDS AND COMPOSITIONS FOR TREATMENT AND PROPHYLAXIS IN THE RESPIRATORY TRACT33
WO2007114881A1- SIALIDASE FORMULATION FOR TREATMENT FOR OF EX PULMONARY DISEASE34
US2008166760A1- COMPOSITIONS AND METHODS FOR ENZYMATIC DETACHMENT OF BACTERIAL AND FUNGAL BIOFILMS35
WO2004066945A2 ENZYMES AND THE NUCLEIC ACIDS ENCODING THEM AND METHODS FOR MAKING AND USING THEM36
US20050112751A1 a novel class of therapeutic protein-based molecules where the use of sialidase is to preventing a pathogen infection37
WO2000010388A1 SIALIDASES AS MUCOSAL ADJUVANTS38
As seen from this analysis, we could not find any patents combining ABC only A, B, C alone or two of them combined. This results in an opportunity to patent ABC together as a system.
A strategic solution after doing the patent research would be to file a provisional patent application during the iGEM competition. This way, you can share all of your work with fellow iGEMers, but still start your own business afterwards.
