Throughout the summer, our goal was to find a suitable way of counting C.Parvum in HCT-8 cells. After trying several dyes, we found that crypt-o-glo and sporo-glo—two monoclonal anti-bodies—worked best in quantifying the parasite during infection.
After finding a suitable way of quantifying the parasite we moved on to the next step in our experiment, which dealt with testing drug inhibitors for an enzyme in cryptosporidium known as SSAT.
Previous experiments conducted at Haskin’s Lab have demonstrated that a drug known as Spermidine/Spermine N1-Acetyltransferase may play a role in parasite survival by controlling the levels of spermine in the parasite. The drug inhibitor used is an analogue of spermine, which is the polyamine that reacts with this enzyme. The inhibitors that we are using are 4-4-4 polyamines, similar to the 3-4-3 structure of spermine and spermidine. This inhibitors inhibit cpSSAT, but not the human SSAT.
We tested these inhibitors against paramomycin, a drug that is currently being used for treatment of cryptosporidiosis, albeit in large doses. Due to this, we hope to find a compound that may be used in low concentrations for the treatment of cryptosporidiosis.
So far, we have tested two compounds by running MIC Assays. During our MIC assays, we found that one compound did not show significant growth inhibition in C. Parvum; however, the second compound showed greater growth inhibition of C. Parvum than paramomycin when compared at the same drug concentration.
We currently still have a library or compounds that will continue to be tested, and hopefully one of these will show greater benefit than paramomycin. After this is completed, we may move to assaying how these compounds may affect human small intestine cells, and in the future, attempt to use these drugs in mouse models for the treatment of cryptosporidiosis.
I have learned a great deal about drug testing, the C.Parvum parasite, and cryptosporidiosis throughout the summer. The labwork, although sometimes tedious and disheartening, allowed for a hands-on environment that further emphasized the importance of my readings and paper research. It has also taught me other skills that can be translated into other experiments.
This project will hopefully lead to a suitable treatment for cryptosporidiosis. Dr. Yarlett’s research has hit close to home for several reasons. One day, I aspire to become a pediatrician, and the fact that many children in third world countries are dying because of lack of treatment is very disheartening. Furthermore; this disease is also threating for people who have contracted HIV, a virus that is very prominent in the LGBT community and underdeveloped countries. I am a civil rights advocate and a member of the LGBT community myself; I have seen the hardships that people with HIV live with, and this disease has been culprit of many of them in the past (such as the outbreak in Wisconsin in 1993) so it means a lot to me that research for this disease is being conducted at Pace and that I am being part of it. I hope that I can continue to work with Dr. Yarlett in order to help create a treatment that will eliminate the threat of this parasite.