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Cancer-On-Chips To Evaluate The Inhibitory Capacity Of Vitamin C On Colon Metastasis To Liver

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In Western countries colorectal cancer represents a significant cause for mortality in both genders. Several genes are involved in genetic alterations such as gene mutations and epigenetic modifications. These genes could led to colorectal cancer by regulating cell mechanisms such as proliferation and apoptosis. The survival expectancy for localized cancer is 5-year. However, for patients suffering from metastasis the survival rates remain less than 30%. Metastasis is defined as the broken off cells derived from the primary tumor, migrate using the bloodstream or the lumph and establish a secondary tumor. According to WHO the majority of patients pass away due to metastasis.

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Considering the several cancer physiological parametres the 2-D cell culture and animals models which have been used so far can’t contribute to metastasis research. At that point, new technology ‘organs-on-chips’ enabled scientists to study for example the migration of colon cancer cells to liver. Regarding the ‘metastasis-on-chip’ for instance fluorescent stained cancer cells derived from colon can be tracked while they are traveling via a device which represents the circulatory fluidic system. The cells travel from the first chip where the primary tumor is located in the hydrogel-fabricated gut to the second chip where liver cells are located.

A well-known mutation which is present in colon tumor is the KRAS mutation. This mutation has a severe impact on the regular cell metabolism. In addition Warburg metabolic is a process which takes place in tumor cells. During this process the glucose consumption rate is extremely high and as consequence there is lactate production. Finally, the interest in high-dose of vitamin C has been aroused among scientists, since it seems that vitamin C in cytotoxic concentrations is able to induce pro-oxidant effects and as a result to kill cancer cells.

Innovative character

Comparing with 2-D cells cultures and animal models, microfluidics represent the new innovative technology solution concerning the cancer experiments with advanced capability of prediction. Microfluidics also allow testing of pharmaceutical compounds. Especially for drug development and vitamin toxicity microfluidic systems considered to be a novel method since represent a vasculature comparable to in vivo properties. Moreover, chips due to their ability to simulate the organs function are a promising tool to study metastatic dissemination and pharmacokinetics action simultaneously. In other words organs-on-chip play a crucial role of appropriate dosages determination regarding treatments and combination of therapies. Furthermore, organ-on-chip provide personalized medicine by conducting drug screening. Personalized medicine is recommended since has already observed that there is extremely high tumor heterogenecity and each patient has different response to different drug. According to the above mentioned, microfluidic devices appear to be a powerful platform for cancer metastasis studies. The Warburg Effect is a fundamental factor of tumor cell proliferation but its function in combination with high dose of vitamin C still remain a challenge. Finally, for vitamin C in known that induce GLUT-1 and (PKM2)-PTB downregulation and afterwards Warburg effect is blocked but more studies should be done.

Experimental design

The study involves 2 participants with colorectal cancer, without having already metastatic liver cancer. Preferably male and female patients. With permission from the medical team and participants, we will obtain part of colon cancer cells after scheduled surgical operation and liver cells by biopsy. Once the cells will be selected will be loaded into chips in order to be cultivated. Six chips will correspond to each patients, three for colon cancer cells and three for liver cells. Afterwards, the chips which represent the colon will be connected to the chips which represent the liver. Colon cancer cells will be treated with 60mg, 120mg and 180mg of vitamin C respectively for each patient. Under fluorescence of the cancer cells, we will monitor the effect of the vitamin C on the metastasis. This will be carried out using a clinical trial design. The examination of the resected specimens will also be processed for genetic analysis in NIRDPBS, where KRAS status will be determined by using two methods: PCR-RFLP and pyrosequencing to check for mutation and metastasis. These chips represent promising models for pharmacokinetics. On-chip 3D in vitro tumor models will be used to investigate the interaction of cancer cells and the subsequent effect of the vitamin C.

Main proposed outcomes

Vitamin C shows certain antitumoral activity, but the molecular mechanism underlying this side effect and the intriguing selective activity displayed is not clear. High amount of dosage of ascorbate may improve symptoms and prolong life in patients with colorectal cancer. The utmost outcome of the study is to know the effects of ascorbate on colorectal cancer metastasis using chip.

The technology organ-on-a-chip has great potential to advance the study of tissue development, organ physiology, disease aetiology and analysis of biochemical, genetic and metabolic activities of living cells as in the case of vitamin C inhibit colorectal cancer metastasis.

Potential for practical application of the envisaged research results in economic and/or social terms

Once the experiment is complete many positive aspects of social benefits should have appeared. First of all economic benefit. It is well known that health science costs a lot of money around the world. Scientists try to treat several diseases. At that point new technologies like ‘organs-on-chip’ contribute to society apart from high accuracy regarding the final predictions and in financial part of experiments. Every single study which will be performed on microfluidics devices will reduce not only the cost but also the time between the experiments and the outcomes.

Nowadays the modern medicine has the duty to respond rapidly since the society needs have increased dramatically. For instance apart from bioterrorism attacks and progressing pandemics the health science has to address and the rapid increase in non-communicable diseases such as cancer. As regards cancer working on ‘organ-on-chips’ help the improvement of experimental techniques by developing new tools. These tools will be able to detect the exchanged between several tissues simulating the whole body physiology and convey metabolites derived from different pathways in real time. Concerning the colon metastasis to liver ‘metastasis-on-chip’ can monitoring and evaluate the interactions between colon cancer cells and their ability to travel and establish a secondary tumor in liver cells taking into account and the dynamic force from bloodstream. To make it more clear this study will narrow the gap which exist so far between the in vitro environmental conditions in 2-D cell culture and in vivo conditions like whole human body. Moreover, using human colon cancer cells on chips scientists are able to avoid animals models. It is well known that there is high time consuming between the experiments which are conducted on animals and outcomes. Additionally to time consuming there is a divergence between animals experiments outcomes and humans experiments outcomes. This divergence can fade away using directly human cell and higher accurancy can be achieved.

Another important point of this study is the impact of high dose of vitamin C on Warburg effect and the process of cancer metastasis. As a result after the performance of the experiment the preclinical stage will be more clear. Last but not least performing this study cancer metastasis from colon to liver is excepted to be inhibited and cancer patients to have a better quality of life.

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