In your future directions you discuss testing the compound with more vials at the same dosage, how would this potentially change the result you previously got?
Testing with a larger sample size would help us minimize outliers in our data (to gain more reliable data for statistical analysis). This would either reinforce what we found in our experiments or demonstrate errors with our sample size in this experiment.
By examining our compound in conjunction with a subclinical dose of colchicine, we would hope to see that this combination of low dosages of both drugs would work synergistically to produce a lower percent survival than we observed by using our compound alone. This could possibly occur by the use of both drugs that have different mechanisms of action. Therefore the combined mechanisms of our compound and another compound would hopefully more successfully result in a lower survival rate.
One possible cause could be logistical issues that we faced with the storage and stability of our compound. Our compound was ideally supposed to be stored at -25 degrees celsius, however this was not realistic for our lab because the drug would have to be warmed to room temperature (or above in the incubator) once the drug was added to the fly food. Therefore, this compound being at a higher temperature could have negatively effected its stability and therefore potency. Another cause of this lacking efficacy, could be that the starting dose that we tested was not able to reach high enough concentrations in the cell to effectively kill those cells.
This trend could have occured for a number of reasons such as incomplete mixing of the food/drug solution or other issues associated with the larvae that resulted in unpredicted trends in the survival rate. If the food/drug was not mixed completely, the amount of the drug ingested by the flies could have been significantly lower than we expected and therefore the percent survival would not follow the expected trends. If there were other abnormalities associated with the larvae that we tested this could also explain this unpredicted trend. If the larvae had some kind of mutation or other health abnormality that either potentially increased or decreased the percent survival separate from the drug concentrations, this could also result in the trends not following our expectations. Ultimately there are many other potential experimental errors that could have resulted in this odd trend and I’m not completely sure why this trend occurred. This is another reason why retesting the compound with higher repitition would be useful.
There are many compounds that have been identified as promising potential chemotherapeutics, both in this lab and in other assays. In our research of our compound, AMP, we found research that showed promising results for using a compound related to AMP (that binds to a different kinase) to treat breast cancer. This compound could be a good candidate for testing, because of these promising results. Otherwise, we could select a drug from the list of potential hits from previous semesters or other drug screens in attempt to identifying a compound that seems to be a good candidate.
Would other kind of environmental factors could have affected eclosing rates? Do you have an idea of how to control for them in a future version of the procedure?
There are many factors that could influence the eclosing rates of the flies. Temperature is one factor that could affect this rate. We incubated the larvae during this 10-15 day period in hopes of being able to quantify sooner (we had issues with setting up our vials late). This exposure to increased temperature could have affected the eclosing rates which would be a useful tactic for controlling the eclosing rates in the future.
In your future directions you discuss testing the compound with more vials at the same dosage, how would this potentially change the result you previously got?
Testing with a larger sample size would help us minimize outliers in our data (to gain more reliable data for statistical analysis). This would either reinforce what we found in our experiments or demonstrate errors with our sample size in this experiment.
What would you expect to evaluate if you combined your drug with colchicine for a possibly synergistic effect?
By examining our compound in conjunction with a subclinical dose of colchicine, we would hope to see that this combination of low dosages of both drugs would work synergistically to produce a lower percent survival than we observed by using our compound alone. This could possibly occur by the use of both drugs that have different mechanisms of action. Therefore the combined mechanisms of our compound and another compound would hopefully more successfully result in a lower survival rate.
Do you have any explanations as to why the AMP didn’t work as expected?
One possible cause could be logistical issues that we faced with the storage and stability of our compound. Our compound was ideally supposed to be stored at -25 degrees celsius, however this was not realistic for our lab because the drug would have to be warmed to room temperature (or above in the incubator) once the drug was added to the fly food. Therefore, this compound being at a higher temperature could have negatively effected its stability and therefore potency. Another cause of this lacking efficacy, could be that the starting dose that we tested was not able to reach high enough concentrations in the cell to effectively kill those cells.
Why do you think your trends for dilutions 2,3, and 4 were not consistent?
This trend could have occured for a number of reasons such as incomplete mixing of the food/drug solution or other issues associated with the larvae that resulted in unpredicted trends in the survival rate. If the food/drug was not mixed completely, the amount of the drug ingested by the flies could have been significantly lower than we expected and therefore the percent survival would not follow the expected trends. If there were other abnormalities associated with the larvae that we tested this could also explain this unpredicted trend. If the larvae had some kind of mutation or other health abnormality that either potentially increased or decreased the percent survival separate from the drug concentrations, this could also result in the trends not following our expectations. Ultimately there are many other potential experimental errors that could have resulted in this odd trend and I’m not completely sure why this trend occurred. This is another reason why retesting the compound with higher repitition would be useful.
Do you know of any potential drugs that may be a better candidate for testing?
There are many compounds that have been identified as promising potential chemotherapeutics, both in this lab and in other assays. In our research of our compound, AMP, we found research that showed promising results for using a compound related to AMP (that binds to a different kinase) to treat breast cancer. This compound could be a good candidate for testing, because of these promising results. Otherwise, we could select a drug from the list of potential hits from previous semesters or other drug screens in attempt to identifying a compound that seems to be a good candidate.
Would other kind of environmental factors could have affected eclosing rates? Do you have an idea of how to control for them in a future version of the procedure?
There are many factors that could influence the eclosing rates of the flies. Temperature is one factor that could affect this rate. We incubated the larvae during this 10-15 day period in hopes of being able to quantify sooner (we had issues with setting up our vials late). This exposure to increased temperature could have affected the eclosing rates which would be a useful tactic for controlling the eclosing rates in the future.