Hello, and welcome back!
It’s my first official week working on my Senior Research
Project over at the Scottsdale Mayo Clinic, and I’m already learning and
growing. Literally. I’ve got myeloma cell lines growing out of petri dishes
like chia seeds on a clay Scooby Doo.
The uncanny
similarities….
Backing up a bit, I began the week discussing our battle
plans against these myeloma cell lines. Armed with some pipettes and a little
bit of trypan blue, the lab has teamed up with a pharmaceutical company in
order to look at other ways to manipulate myeloma. We have decided to test
antisense oligonucleotides (ASO’s), which attach to mRNA to stop its
manufacture of protein, specifically for myeloma, IRF4. Recent studies have
shown that IRF4 may be the “Achilles Heel” in the Trojan war against myeloma. Myeloma
seems to feed off of the production of IRF4, and an arrow to the production of
IRF4 may kill myeloma cells entirely without harming normal cells. So, the
question remains, can ASO’s be this arrow? And, will all types of myeloma be
subject to the predicted effects? If not, why?
With these technical plans set firmly in the future, my
present is momentarily absorbed by the certainty of two things:
1. The
equation (M1)(V1)=(M2)(V2) is not to
be underestimated
2. Pipetting
requires serious deltoid muscles
Despite my aching arms, working at the lab has definitely
been the highlight of my week. It’s been a whirlwind of warming media, using
acronyms, looking under microscopes, and being totally amazed by the
advancement of science.
I hope to see you again next week!
Until then, thank you and happy reading!
-Angela (or “The (M1)(V1)=(M2)(V2)
girl”)
Hi Angela,
ReplyDeleteSince you are The (M1)(V1)=(M2)(V2) girl, can you explain it in a way that an English major like myself could understand?
Mr. Bloom
Sure thing Mr. Bloom!
DeleteIt's a simple equation to determine concentrations and volumes. It is particularly helpful with dilutions. So, for example, if we had a certain concentration of a drug (lets say it's a concentration "A") but we want to have a smaller concentration of it (perhaps concentration "a") in a certain volume (predetermined volume "B"), then we would use the equation to find what volume of the higher concentration drug to use (let's say volume "x"). So, the equation would look something like this:
(A)(x)=(a)(B)
So, we'd find x with some simple algebra:
x= (a*B)/(A)
And then, in the lab, we'd pipette out x amount of "A" concentrated drug and dilute it so that it became "a" concentrated drug.
Hope that helps!
I enjoyed your comparison at the beginning of the post. I was wondering why the ASOs work, though. How do the stop the manufacturing of the proteins? Also, can you maybe provide some more background on IRF4?
ReplyDeleteHey Lauren!
DeleteThank you!
ASOs can be engineered so that they are the compliment of a certain genetic sequence. In this case, the ASOs are matched to the mRNA that would usually be translated into the protein IRF4. So, the ASOs will bind to the mRNA, stopping the translation of mRNA, and halting the production of IRF4.
As far as IRF4 goes, it is a transcription factor (a protein that binds to DNA in order to begin the replication of DNA). IRFs are normally seen in lymphocytes and play an important role in immunology. However, in multiple myeloma, IRF4 becomes overexpressed. This is fairly new research, and we still don't have all of the information. But I hope that this has helped you a little!
I am right with you with the equations and pipetting. This looks like it will produce some interesting results!
ReplyDeleteAngela, I think the highlight of my week is going to be reading your blog. I expect a comparison of no lesser value than this one next week! Oh, and I'm also excited to hear about your findings!
ReplyDeleteThanks Val! No promises, but I just might mix it up from chia to sea monkeys...
DeleteHey Angela, very interesting topic and entertaining post!
ReplyDeleteCould you talk a little bit about the specific tests you're using? Since you're using trypan blue it sounds like you might be doing a cytotoxicity assay.
Hi Navya,
DeleteThank you! And yes, that is exactly what I'm doing. However, at the time, I was using the trypan blue just to count the cells. But the experiment will also be tested through multiple types of assays.
Angelina....this is so exciting!! With all this pipetting, you may be giving Zachary and Roshan a run for best deltoids! Seriously though, while I and others from bio can follow the strategy of the lab, you might have a challenge here to make what you are doing more understandable to those whose molecular bio is a tad bit less extensive than yours. This is going to be a fun SRP to follow
ReplyDeleteThe metaphors made this easier to understand (and more readable) than I expected from something that could otherwise be really dense and obfuscated by medical jargon. Thank you for that. This project sounds really useful and I'm excited to see your results!
ReplyDelete