-4-year-old goofball who loves superheroes, school, and her best friend Lincoln.
-Nat G was born in 2017 and was quickly deemed a normal but “floppy” baby who would grow out of that “very soon”.
-At about 12 months old, we started physical therapy because Nat G didn’t seem to hit many “gross motor” milestones after sitting up.
-Fast forward to almost 2 years old and she was testing at 5-6 years old intellectually and with her language (basically we couldn’t shut her up! ha!).
-She was still stuck at 9 months physically.
-In Early 2020 just shy of 3 years old, and after a few layers of genetic layers.
Myopathies are a group of genetically heterogeneous conditions characterized by muscle weakness, with overlap in the clinical presentation and histopathological features of different genetic subtypes.1 Within this group, congenital myopathies are most commonly characterized by hypotonia and weakness, often from birth, commonly with the presence of facial weakness, with or without ptosis and ophthalmoplegia.
PYROXD1 gene homozygous mutation is a very rare disease that appears to exhibit a LGMD-like phenotype with childhood or adulthood disease onset. It has few additional clinical features including scoliosis, high arched palate, nasal speech, and joint hypermobility. Awareness of these clinical features may be helpful to facilitate the diagnosis and lead to the appropriate genetic testing.
PRYOXD1 is an oxidoreductase – which sounds complicated, but just means it can oxidise and reduce ‘things’. When we discovered PYROXD1, there was not a single science paper published on it, ever. We have been working from ground zero to try and find out what it does. We know that no other enzyme can reduce/oxidise ‘the thing(s)’ that need PYROXD1 – and whatever the things are, they are absolutely critical, because cells and mice do not survive without any PYROXD1.
The team of doctors and researchers were the first group to discover PYROXD1 as a novel disease gene causing myopathy in five families around the world. O’Grady et al, American Journal of Human Genetics 2016. They are aware of almost all cases around the world. They have been studying PYROXD1 for 7 years now – trying to find out what it is that PYROXD1 does that humans need so desperately and working our way toward a therapy.
PYROXD1 (Pyridine Nucleotide-Disulphide Oxidoreductase Domain 1) is a Protein Coding gene. Diseases associated with it present as extremely rare and progressive forms of muscular dystrophy. Less than 20 people worldwide have this specific mutation, one of those people is our daughter Natalie. Her case is considered extremely severe, as her symptoms set in at a very young age. Natalie cannot walk, crawl, or do any of the things any other 3-year-old can do. She can barely use her muscles at all. A simple cold means endless trips to the emergency room. Natalie needs constant round-the-clock care, receives hyperbaric oxygen therapy twice a week ,and uses a G tube to eat. She fights every day.
Her mutation has no cure. And, due to the small number of cases, traditional research funding from pharmaceutical companies and others simply doesn’t exist. It doesn’t make financial sense.
Funding for this project, estimated at $280,000, will allow researchers to discover insights into the origin of this mutation, how it affects the body and possible treatment options. We know this won’t save Natalie, but we hope it will lead the way for finding a cure for PYROXD-1 and help others with rare pediatric diseases.
The researchers are interested in the 3D structure of PyroxD1 with and without it’s ligands (NADPH and it’s natural ligand). They use X-ray crystallography and cryoEM to look at protein molecules. The hope is that they will understand the enzyme better, and we can use that information to design more effective treatments. One of the most promising parts of this project is that the researchers have been able to make as much of the enzyme as needed so they are also pursuing potential enzyme replacement therapy with doctors ranging from the United Sates of America to Australia.
With the funding received from the project, the researchers will be able to get preliminary data on their theories about PyroxD1. Once we get that data together and published, we can write federal NIH grants that can be more directed for therapy. This funding will help the gather the date needed to be able to be published. The findings along the way are likely to also have broader impact on other rare conditions due to the level of unknown when we are talking about the specific gene.
The ideal outcome of this project is to to use the researchers structural data to design drugs that may compensate for any mutations in the natural PyroxD1. A similar approach was successful with cystic fibrosis, but they needed a 3D structure of the cystic fibrosis chloride channel first.. In addition, successful enzyme replacement therapy would be a great outcome as well. This would help not only the few people diagnosed, including our worrier Nat G but also the countless people who have this mutation but were not able to be diagnosed yet.
This research is incredibly valuable due to the amount of unknown about these specific gene. The research can give us insight to not only this gene, but other ways to research unknown genes as right now there is more we do not know about genetics than there is we do!
Directly related to this project, the researcher would be able to expand on the currently available structural information that is available for PyroxD1, and to use this information to discover ligands and enzyme activity-compensating therapeutics. To start that sort of project, they really need to search for the natural ligand for this enzyme using commercially available screens. These screens are large collections of chemicals that should be present in the same area of the enzyme; therefore, one of them is probably a natural ligand.
If we do not secure the needed funded, the researchers would have to cease everything. Most American labs operate on a small-business model and like any small business, they need to find a way to get working capital. Unlike a small business, they are not selling anything, they are searching for something. this group has already had to get research grants to pay for as much of our their own expenses as they can, in addition to the labor for those working on these projects and the supplies/services that are require. That includes the electricity, water, heat, and air conditioning, Administrative staff, etc.…Without funding, all that stops.
After we get the money, these researchers can stop worrying if the money will come and instead can just focus on this project. They would be able to cover all expenses related to these test as well as allow them to collect sufficient data to write a full NIH grant. By getting this project funded and continuing to move along, these researches are confident in what they will find and then be eligible to receive federal support and likely get 5-years of steady funds that will be used to address the ideas that we propose.
Time Frame – 2 years of Research
Budget – $280,000
The researchers choose to support are responsible for the direction of where the funds are going. Below you can see the benchmarks our researchers intent to hit for each $25,000 we are able to donate for along with what they intent to use funds to do at that point. The following is based on the researches intent to:
$0-$25,000 – Secure United States personnel and consumables to produce large quantities of recombinant PYROXD1 enzyme.
$25,000-$50,000 – Complete pre-clinical evaluation of potential therapeutic benefit of PYROXD1 enzyme replacement therapy using Cell Models.
$50,000-$75,000 – Acquire mice in Australian laboratory to be used for the next step (Mouse Models).
$75,000-$100,000 – Progress from the findings in the cell models to pre-clinical evaluation of potential therapeutic benefit of PYROXD1 enzyme replacement therapy using using Mouse Models.
$100,000-$125,000 – Develop necessary personnel in Australian laboratory to assist in the research of PYROXD1 and reviewing, collecting, and recording all findings.
$125,000-$150,000 – Obtain reagents (PYROXD1 enzyme) for the Australian laboratories to be able to continue studying.
$150,000-$175,000 – Secure equipment needed to better evaluate latest findings for PYROXD1.
$175,000-$200,000 – Develop personnel in Australian laboratory to assist in the research of YROXD1 and reviewing, collecting, and recording all findings.
$200,000-$225,000 – Creation of more reagent (PYROXD1 enzyme) at the United States laboratories.
$225,000-$250,000 – Compile all findings in the models above (specifically the mice modal) to be use and apply for a much larger grant to help take these findings and prepare for potential clinical trials.
$250,000-$280,000 – Organize data from the findings and apply for needed grant to bring these findings into a therapeutic option.
Three Ways You Can Take Part
The researchers Take-Part chooses to support are responsible for the direction of where the funds are going. so the plans above have been designed by the researcher with no influence from any donors. When it comes to medical research, we need the researchers to be able to pivot as needed in their research as findings are discovered. The above is what the researchers have shared they intend to use funds raised to achieve the desired goal of this project. Take-Parts role in this is to help caring donors like you, put funds towards amazing researchers that we decide to support, and allow the researchers to do what is best for the progression of this project.