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Whether you are a fresh cancer research scientist or a senior expert, the idea behind designing an outstanding and publishable original research remains the same if we are talking about the basics of developing and designing research projects. In fact, it is way easier than you would think. In this blog, I will guide you through the steps to easily build your own project if you are thinking to run your research for the first time or already received your research grants or even if you are on a tight budget. However, the more you get familiar and digging deeply in the project, the more complicated things will pop up at one point and only your experience would allow you to handle and complete the road map. It is indeed that this point is actually what distinguishes fresh from well-experienced researcher; solving problems and finding solutions for complex questions. It is also worth mentioning that this strategy does not differentiate between different goals of your project. That means whether you are evaluating cancer candidate drugs or study mechanism of cancer progression or even fishing potential biomarkers for diagnosis or prognosis, it will remain the same direction as long as you are conducting preclinical cancer research projects. 

1. Start by establishing clinical effect and clear observational findings (Go In vivo before in vitro!)

First, determine which model is the best for your project. Think about different models of cells/ mice, that are in literature such as orthotopic, transgenic, surgical, xenograft etc. and make your modifications according to your demands. Think always about the reproducibility of your model! Secondly, Let’s assume that you have a potential pro-drug, antibody, inhibitor, or even your own potential extract. The most important part of your project will be relied on this attempt. In fact, this also would strengthen your application if you are applying for grants and you have included some pilot or preliminary results. At this step, it is very crucial to establish clinical findings before you start your in vitro experimentation as this would save a huge amount of money and will direct you where exactly to go. Do test your material in vivo and see if the desired effect has been achieved.

Once you are happy with the findings, let’s say you get a nice tumor regression in the animal model, then it is time to go for in vitro digging. To sum up, your initial steps briefly, establish the effect first and then solve your puzzle!

2. Which experiments do I need to evaluate the In vivo findings?

Next, it is time to investigate around your in vivo findings. Tumor regressed? That’s nice! Think about the predictable mechanism. Is it because of tumor proliferation, or tumor cell metastasis, or increased apoptosis, or could it be the tumor microenvironment? Which potential genes are involved in each step? Is there any effect on using different doses? If you are able to address these questions to your model, then BINGO! You are almost done. What you need basically according to the previously mentioned questions is proliferation assay, tumor metastasis assay; maybe migration assay or in vivo imaging. Apoptosis assay is obviously required to evaluate the selectivity of your drugs. Isolate tumor microenvironment cells and study each component according to their ratio. Which lineage is of particular interest? This depends on your model. It could be macrophages that enhance metastasis or T reg cells that help tumor escape from immune surveillance or Neutrophils that aid in tumor progression. Gene and protein expression studies are also essential in identifying biomarkers and pathways of regulation. 

3. What technologies do I need to assess the identified studies?

Again, if you are able to address the previously mentioned studies, then BINGO the technology you want is just right on the corner. PCR and western blot are required for gene and protein expression. Perhaps you would choose ELISA instead of western blot if you are interested in quantitatively determination of your targets. Flow cytometry another potential resource for evaluating surface and intracellular protein expression. There are also wide variety pf detection panel of Proliferation and apoptosis; BrdU, DNA staining, Annexin V, Fluorescence molecules such as Calcein Am, Persto blue, caspases, etc. all these arrays can be formatted either fluorometric, colorimetric, or even by immunofluorescence meanings for instance flow cytometry or immunofluorescent microscopy.

4. Go back again to your In vivo model!

Now given that you gained some insights about the mechanism, or fetched biomarker, or assessed your drug, it is now time for real dramatic experimentation. Why? Through your previous studies, you should gain some understanding of your target. Now you need to translate the in vitro findings to in vivo though it is not always the case. For example, if you have found your happy biomarker then it is advisable to find a way to overexpress it in your system to evaluate what so-called gain-of-function or loss-of-function studies that would eventually judge your overall model performance. As I mentioned earlier, things might cross your way and required thorough investigations. Having said all the above, you are ready now to collect your data for analysis and find your journal of interest to publish. Should you have some tips on how to write your scientific manuscript, I would be glad to share with you some tips in my next blog. This is the way IF everything went smooth for a straightforward preclinical cancer research project.

Finally, make sure always your data are reproducible and high quality and believe me, the more research you do independently, the more experience you will acquire in running and managing the core fundamentals of cancer research.

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