Disease States and Potential Therapeutics
Youhai Chen, M.D., Ph.D., associate professor of pathology and laboratory medicine at the University of Pennsylvania, says that miR-21 is “probably the most famous microRNA there is.” miRNA-21 is an oncomiR; just like an oncogene, it is upregulated in a number of cancers and promotes uncontrolled cell growth. But Dr. Chen studies its involvement in type 1 diabetes. miR-21 targets the tumor suppressor PDCD4 (Programmed Cell Death Protein 4).
In lymphoma and leukemia, elevated levels of miR-21 promote oncogenesis by preventing the translation and accumulation of PDCD4. Dr. Chen and his colleagues have found that the same is true in pancreatic β cells, the insulin-producing cells that are killed by the immune system in type 1 diabetes: elevated levels of miR-21 suppress PDCD4, and can thus prevent the death of these pancreatic β cells.
Dr. Chen will be presenting this work at CHI’s conference on “microRNAs in Human Disease and Development” in Cambridge, MA, next month.
Matthew Evans, Ph.D., an assistant professor of microbiology at Mount Sinai School of Medicine, studies hepatitis C (HCV). This is difficult to do because the virus doesn’t replicate well in culture. Only one cell line can support it, and that cell line is far from ideal because it is not polarized like the hepatocytes HCV actually infects.
miR-122 is a liver-specific miRNA that regulates cholesterol and fatty acid biosynthesis. “Instead of inhibiting gene expression like most miRNAs, miR-122 actually promotes replication of the HCV genome by directly interacting with the far 5´ end of the viral RNA genome. This is pretty much the opposite of a normal function of a microRNA,” noted Dr. Evans, who will also be presenting at the CHI meeting.
This phenomenon was first discovered in 2005. In 2011, Dr. Evan’s lab showed that, when put into polarized HepG2 cells along with a required HCV receptor, miR-122 can support the entire viral life cycle, including infectious virion release. “This doubles the number of cell lines we can use to study HCV,” Dr. Evans says.
Interestingly, the stabilizing action of miR-122 does not require the viral RNA to be transcriptionally active, and miR-122’s normal function is not required either. Sequestering miR-122 may potentially be an effective anti-HCV therapy. miR-122 has been successfully silenced in African green monkeys, and the treatment is now in Phase II trials.
Just down the hall from Dr. Evans, classically trained virologist Benjamin tenOever, Ph.D., set out to determine if miRNAs play a role in the cellular response to viral infection. Alas, they do not.
But Dr. tenOever persevered; he figured, “Just because miRNAs aren’t antiviral, doesn’t mean they couldn’t be!” His lab then generated a flu vaccine by grafting an miRNA target site into the influenza virus. He chose an miRNA that is present in mammalian but not chicken cells, so the virus grows perfectly well in eggs and is only attenuated when it infected mammalian cells that endogenously express the miRNA. Unlike canonical miRNA target sites, which are not perfectly complementary to the miRNA, Dr. tenOever made this target perfectly complementary.
The miRNA thus acted more like an siRNA, attenuating viral replication 1,000-fold instead of the two- to threefold usually achieved by miRNAs. And unlike other flu vaccines, this one does not rely on the immune system to destroy it, so it can be administered to immunocompromised individuals without eliciting unpleasant side effects.
Dr. tenOever has played with engineering miRNAs and their target sites into other viruses as well, generating viral vectors targeted to specific cell types—much like HCV evolved to replicate only in hepatocytes because of its reliance on miR-122, which is expressed only in those cells.
He has also made viruses that can churn out an miRNA of choice once inside cells. “We don’t work with viruses and miRNAs in a conventional sense,” says Dr. tenOever, “but we do a lot of things with them, and our toolbox for molecular therapeutics has grown because of it.”