Team:GreatBay SCIE/Safety

Safety



General Safety

Our laboratory ,where most experiments are done, is classified as Biosafety Level 1. This means our laboratory settings in which personnel work with low-risk microbes that pose little to no threat of infection in healthy adults. Our laboratory consists of research taking place on benches without the use of special contaminant equipment. At the same time, even though a BSL-1 lab is not required to be isolated from surrounding facilities and houses activities, our laboratory is separated into several zones with walls.（Not shown in the picture; this area is dedicated to iGEM teams)

In addition, before enrolling in the team, each of us was required to pass an exam including general lab protocols and lab safety. A formal handbook which consists of general lab protocols and lab safety is handed to everyone and pasted on wall as well. Before conducting experiment, we are required to wear lab coat and gloves; taking off them, dumping gloves in trash bin and not taking anything out lab is also required.



Project Specific Safety



Use of doxorubicin

Doxorubicin is a drug enveloped into nanoparticles for treating breast cancer. After communicating with Dr.Zeng from Southern University of Science and Technology, we have learned that doxorubicin can cause serious damage to our health, including cancer. Therefore, we strictly follow lab safety rules such as wearing masks, gloves and lab coats. The whole experiment is done within fume hood ,and is under the instruction and supervision of Dr.Zeng.



Preparation of nanoparticles

During the preparation of the nanoparticle, chlorofrom was used as an organic solvent to dissolve PLGA. As a solvent not frequently used in our lab, we paid special attention to its hazard and we have inquired Dr.Wei about the necessary procedure required when working with it. We were informed that the solvent causes irritation to skin and eye; is toxic if inhaled and suspected of causing cancer. Dr. Wei stressed that gloves and goggles must be worn and most importantly, experiment operation involving chloroform should only be performed in fuming cupbord. We were supervised throughout the experiment and rules suggested by Dr. Wei were strictly followed

Cell Culture

We have been culturing cancer cells in order to test the effectiveness of our system. The chemical(DMSO) used to culture cells and cells both constrict a potential threat. To protect ourselves and cells, we conduct the whole experiment in Shenzhen University, under the help and supervision of Miss Cao(a student of Shenzhen University)Moreover, we strictly follow safety protocols incorporating sterilizing ourselves with ethanol sprays; sterilize the whole laboratory with UV light. In addition, considering the safety issue of cancer cells, we clear out one separate chamber of incubator for storing cells.


Figure 4: Age distribution of patients with breast cancer Figure shows age distribution in China and the USA in 2008, and estimated distributions in China in 2020 and 2030; based on data from the WHO China country profile.22 Many effective drugs have to be imported overseas which is much more expensive and not included in the national medical insurance. The fast-pace-lifestyle in China's mega city have also caused breast cancer to arise in younger ages. In big cities like Shenzhen, young people face a lot of pressure and work long hours; as a result, they not only have higher chances of getting breast cancer, but also miss out on regular check ups in hospitals.


Poverty-limited treatment

In China, each period of chemotherapy costs around eighty thousand if using the imported drug with less side effect and higher chance to cure the disease. Therefore, the whole course requires that family to use about 200000 RMB depending on the severity of the disease. Most of the time, the diseased family member can't be at work considering the health condition, leading to the lack of household income, even family financial crisis. Because of this, some women goes to work right after the chemotherapy despite syndromes like constant nausea and dizziness, just because she has to earn money to supply her treatment. If domestic drugs are applied, the government provides medical insurance that pays for 70% of the expense, releasing some of the pressure;however, this policy has not been implemented by all cities around the nation. The economic stress also leads to different trust issue between family members, resulting in a more serious level of disease.


Current Drugs.

Newly trending trending targeting drugs like Antibody-Drug conjugates have proven to be much more effective then traditional drugs, but they are much more expensive (costs around 54,000 US dollar in China), and they only target HER2 Positive breast cancers (which only accounts for 20-30% of breast cancer cases). Currently, there are only two ADC drugs approved by the FDA (T-DM1 and DS8201-a).

  • Long production period -> Expensive
  • Heavy side effects
  • Narrow range of receptors can be used which causes less drugs to be used.

ONCOKILLER

Our project aims at utilizing aptamers to develop a new type of breast cancer targeting-drugs. Compared to targeting-drugs whose specificity was provided by antibody, our project uses aptamers as guide molecules, which provided us with advantages such as lower production costs and shorter production period. See more in Design.

1. Short Production Period
Development of antibodies involves several immunizations of a host animal, isolation of antibody-producing cells, and fusion with fast-replicating myeloma cells. This is followed by hybridoma selection and antibody production. Each step is time-intensive, with the entire process realistically taking four to six months. Aptamers on the other hand, uses SELEX process to be developed which usually takes 2-3 months and may even be short depending on the the aptamer. The actual production of the aptamer can be completed in just a few days.
2. Versatile
Antibodies are confined to a narrow range of receptors while aptamers can be used to target almost all molecules ranging from ions to virus, and tissues. Different types can be developed using variations of the SELEX protocol.
3. Weaker Side effects.
Unlike Antibody Drug Conjugates, our drugs are enveloped by nanoparticles which can increase the drug's stability during circulation in the human body and preventing it from contacting with the normal body cells. Another implementation is pH sensitive aptamers that only bind with the target at low pH, which reduces the chance of binding to most normal cells. Cancer cells have lower pH thus will 'attract' more drugs to their proximity

Looking into the Future

Although ONCOKILLER seems prosperous. We have countered several issues during our time in the lab, which you can read about in our Results page. However, there is more to it than this. If ONCOKiller can be refined and optimized it could become a universal model for many cancer drugs. Due to the accessibility of aptamers and its versatility, we believe that more cancer curing drugs can be made cheaper, allowing more families to be able to afford them.