NACFC 2017:Moving Gene Editing from Science Fiction to REALITY.

Gene editing and mRNA editing have been discussed at NACFC for a few years now. Here is the link to last year's blog and conference slides. Researchers have been working hard to overcome obstacles to delivery and expression of genetic treatments over the course of the last year...and they are getting somewhere. I spent Friday afternoon, as well as Saturday morning listening to talks about gene editing. Check out this animation illustrating what gene editing might look like in cystic fibrosis.

As seen in the video, nucleases can be engineered to create site specific, double strand breaks at desired locations in the genome. Antisense Oligonucleotides can be thought of as our "molecular scissors" that guide the gene editing process.

A First-In-Human, Phase 1B, Dose Escalation Study of QR-010, A Novel Antisense Oligonucleotide Administered in Subjects with Cystic Fibrosis Homozygous for the F508Del CFTR Mutation. Elborn, S.

Pro-QR is one of the companies really leading the way with gene editing technologies in cystic fibrosis, and this was a highly anticipated discussion, as this is an early gene editing trial in HUMANS WITH CYSTIC FIBROSIS. 

Pro-QR's treatment is called "QR-010"

An "antisense oligonucleotide" is a synthesized strand of nucleic acid that is complementary to the gene's messenger RNA.
Pro-QR's treatment was administered to the lungs via nebulizer.

Goals are to establish safety and tolerability, as well as assess maximum tolerated dose (MTD).

CFQ-R= Cystic Fibrosis Questionairre-Revised, RSS= Respiratory Symptom Score

Quality of life improvements were patient reported using the Cystic Fibrosis Questionnaire-Revised (CFQ-R), and the Respiratory Symptom Score (RSS)

Wow! Just Wow!!!!!!

Antisense Oligonucleotide-Mediated Upregulation of CFTR via Steric Block of Post-Transcriptional Control Elements: A Novel Approach for Cystic Fibrosis Therapeutics. Sasaki, S.

Antisense oligonucleotides function as our "molecular scissors."

Antisense oligonucleotides are programmed to target a specific site.

Could this type of treatment be successful for gene transfer in CF lungs?

ASOs = antisense oligonucleotides (our molecular scissors).

In Utero Nanoparticle Delivery for Site-Specific Genome Editing. Ricciardi, A.

In 2015, researchers from Yale published a study in Nature Magazine where the F508del mutation was corrected in a living mouse using their specially engineered "peptide nucleic acid" (PNA) to edit the genetic defect. 

“What the PNA does is clamp to the DNA close to the mutation, triggering DNA repair and recombination pathways in cells,” Egan explained. 

Perhaps even more importantly, the Yale group has developed a unique way to deliver their genetic treatment using nanoparticles--particles less that a billionth of a meter in diameter--designed to penetrate targeted cells. Encouraged by their success in living mice, the research group decided to try to deliver their treatment to a developing mouse embryo with cystic fibrosis. Imagine a world where cystic fibrosis could be cured before a child is even born...

The CFF registry has confirmed that early diagnosis and intervention leads to better health outcomes.

We have the ability to detect CF in developing fetuses.

The fetus is deeply affected by cystic fibrosis--even prenatally.

Could in utero gene editing PREVENT cystic fibrosis altogether?!?!

Stay with me here! Remember, PNA is our pair of "molecular scissors," programmed to make the cut at a specific target.

One of the most important things about this research is the FORMULATION of their genetic product. They do this special "nanoparticle" thing--where they cram all the genetic info onto these itty bitty spheres. 

The PNA/DNA nanoparticles work to correct the F508del mutation in living mice.

So they decided to test their treatment on a developing CF mouse embryo...

Holy crap, is that a mouse uterus?!

Researchers have to use specially made glass pipettes to inject their nanoparticles into this particular vein in the uterus.

So cool. This is the actual injection of the nanoparticles into the uterine vein.

Researchers were able to show uptake and expression of desired genetic information in fetal organs!!!

In conclusion...holy shit. This talk left my jaw hanging open!

Manipulating Vector Interactome to Enhance Gene Delivery. Siefert, M.

As previously noted, perfecting the "vector" to enhance gene delivery was a hot topic because it has proven to be so challenging over the years.

Researchers have encountered multiple obstacles to success gene transfer.

Another example of nanoparticle delivery. Could this be the way forward?

Safe, but not perfect.

How do we improve efficacy?

Penetrating mucus is a formidable barrier in CF.

Manipulation of other cellular interactions can enhance gene transfer.

It is worth taking a closer look at how to achieve maximal benefit from manipulation of these cellular interactions.

Researchers were able to show improved efficacy of their gene product by manipulation of cellular interactome.

What an amazing world we live in. Cure Cystic Fibrosis.