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We are excited to share with you results from a recent experiment that demonstrate the efficacy of GALYA-1 DNA in human cells. Read more below!

Gene Therapy-Treated Cells Make Functional NPC1 Protein and Reduce Cholesterol Storage

Researchers at the University of Pennsylvania (UPenn), in collaboration with Galyatech, used human NPC1 fibroblasts (skin cells) to test the efficacy of GALYA-1 DNA.

Why and how did we do this? Before we manufacture GALYA-1 on a large scale, it is important to show that GALYA-1 DNA 'works' – ie. that when we insert it into a human NPC1 affected cell, the cell starts to produce functional NPC1 protein, which it didn't do before the DNA was added.

In the UPenn experiment, some cells were given both GALYA-1 DNA, which contains a copy of the human NPC1 gene, to treat the disease, and green fluorescent protein (GFP) DNA, which makes the cells bright green, to identify which cells got GALYA-1 DNA. After treatment, many cells showed NPC1 protein and GFP expression that coincided with low levels of stored cholesterol, which indicates that the NPC1 protein was functional. This is a good therapeutic result.

In contrast, cells that were not treated didn't make enough functional NPC1 protein to prevent cholesterol storage. This is a well-known negative consequence of NPC1 disease.

These results clearly demonstrate the efficacy of GALYA-1 DNA in human fibroblasts in vitro: treated cells made functional NPC1 protein and cleared out stored cholesterol. You can see the proof for yourself, below.

Microscope Images

To help visualize these results, the following images show the same three human NPC1 cells: one treated (top-left) and two untreated (bottom and right). In the cells,

green is GFP,

red is NPC1 protein, and

blue is stored cholesterol (filipin staining).

Image G1: Green, Red, & Blue

In image G1, you can see that cell #1 is bright green and red; this cell was treated with GFP DNA and GALYA-1 DNA. Cells #2 and #3 were not treated with any DNA and are blue. These two untreated cells lack green or red color, which indicates there is no GFP or NPC1 protein in them. In contrast, the treated cell (#1) is brightly green and red indicating strong expression of GFP and NPC1 protein.
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Image G2: Red & Blue

In Image G2, the green color has been removed to show the extent of NPC1 protein expression (red) in the treated cell (#1). Again, the two untreated cells (#2 and #3) lack red color indicating no NPC1 protein, while the treated cell is bright red indicating expression of NPC1 protein. This is a good result, therapeutically.
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Image G3: Blue

Image G3 is the most exciting of the three because here we have visual confirmation that not only do cells treated with GALYA-1 DNA produce NPC1 protein (red color inside cell #1 in the previous images), but more importantly this NPC1 protein is functional! By removing both the red and green colors, we see only the cholesterol (blue, filipin staining). The two untreated cells (#2 and #3) have bright blue color indicating lots of cholesterol storage, while the treated cell (#1) shows very little blue color, which indicates that the NPC1 protein performed its function of transporting cholesterol out of the lysosomes! This cell shows very little stored cholesterol and therefore we can be confident that the cell made functional NPC1 protein when treated with GALYA-1 DNA.
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Developing GALYA-1

We continue to push forward with GALYA-1 development, including the preclinical mouse and cat studies and manufacturing. Galyatech remains committed to developing GALYA-1 as a safe and effective drug – including initiation of a Phase I clinical trial – as quickly as possible.