Team:KAIST-Korea/Project/Introduction

Why E.canD?

  Two fifths of mankind is suffering from cancer during full life-time, and one fourths dies because of this disease. The rate of people who get cancer increases with age. However, human cannot notice easily about getting cancer or not due to incubation period. Doing so is easier when cancer’s diagnosed at an early stage as treatment is often simpler and more likely to be effective. So finding cancer early is very important. Half of people are killed by cancer within five years after a diagnosis from doctor, and cancer shortens 15~20 years of lifespan of patients. Another interesting research said that 40% of cancer patients experience severe mental distress; most of them are suffered by pusillanimity. It can make one's life tragedy, not only oneself, but also one's family. Another terrible cause why cancer is severe to us is medical expenses. If someone got a cancer, we will receive huge doctor's fee like hundreds of thousands dollar. It makes terribly huge burden for ordinary family. Unfortunately, someone is sometimes suffering to cancer without any treatment. In the case for women, breast cancer is really severe cancer and the most frequent cancer that can find. The following graph shows that a quarter of women who got a cancer have a breast cancer.

  There are several methods for cancer diagnosis. One of the widely used methods is imaging test. It is divided into in vivo and in vitro method. In vivo imaging tests are X-ray, MRI, CT, PET, SPCT scan, and Ultrasound, in vitro method is biopsy. A biopsy is a procedure to remove a piece of tissue or a sample of cells from body for laboratory analysis. In addition, the diagnosis methods using bio marker such as RNA and DNA are developed. Bio marker plays a key role in risk assessment, assessing treatment response, and detecting recurrence and the investigation of multiple bio markers may also prove useful in accurate prediction and prognosis of cancers.

  We focused on the method with bio maker, especially proteins, which distinguish between normal and cancer cells using expression level difference. Many researchers have chosen antibody to detect these kinds of protein, and it needs special luminescence material to confirm whether antigen (expressed protein) is stacked or not. However it is too complicate and delicate process to attach luminescence substance to antibody and it takes lots of cost and time. In addition, established diagnosis method causes social problem because of gap of demand and supply. For these reasons, we have constructed another accurate and rapid cancer diagnosis system using bacteria. We made bacteria to synthesize fusion antibody and to fix it at the membrane itself and by cognizing antigen bacteria emit fluorescence. We want to establish new diagnosis method with synthetic biology.

Whole Pathway

Flash

1. 'E. canD' is spread on the cancer cell.
2. E. canD’s antibody combines with the antigen on the cancer cell.
3. Antigen-antibody interaction make phosphorylate each receptors.
4. JAK autophosphorylation activates transcription factors called STAT1.
5. The activated STAT1 dissociate from the receptor.
6. The activated STAT1 form dimers.
7. The STAT1 dimer regulates GFP gene that attached to GAS elements of promoter.
8. 'E.canD' expresses GFP.

Antibodies for Detecting Cancer Cells

  Cancer cell has distinctive protein expression pattern. This expression pattern also varies on the cell line of cancer cell. For example CD15 and CD30 proteins are OVEREXPRESSED on Hodgkin’s disease, kind of lymphoma, and CEA is over-expressed on adenocarcinoma cell. These protein expression patterns also vary on the cell line of cancer. For example breast cancer cell has different protein expression pattern; over-expression of Estrogen receptor, Progesterone receptor and KI67 of MCF-7 cell line and over-expression of Human Epidermal Growth Factor receptor, KI67 of AU-56 or SK-BR-3 cell line. Therefore, it allows us to know not only the existence of cancer, but also the type of cell line to identify the protein expression pattern of biopsy sample. We selected Estrogen Receptor and HER2 for our project to make a diagnosis the breast cancer. The reason why we selected these proteins is that they are differently over-expressed. Comparing over-expression pattern of these proteins can diagnose existence of cancer and type of cancer.

For Making Biosensor – E. coli Cancer Detector

A biosensor is a device for the detection of an analyst that combines a biological component with a physicochemical detector component.[1] In order to make biosensor with genetically engineered organism, we should consider the following basic sensing process.

The ‘Receive input’ part can be alternated by represented receptor, the ‘Process’ part can be signaling pathway which changes input to output, and the ‘Extract output’ part can be something easily detectable by human. Nowadays, pigments or fluorescence proteins are usually used for output, and in order to express these substances, gene expression is made constant use in genetically modified organism. Thus, all engineered biosensors with organisms should be modeled on (consist of) 1) biochemical receptors which can activate gene expression, 2) biological pathway, and 3) gene activation for output.

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