Considering a research GoFundMe...

Well, my 14 years of working for a lab are almost over... and, naturally, we get the announcement that the hospital is shutting down our little lab right as I'm close to making a breakthrough in craniofacial developmental genetics. I believe I've found the genetic pathway AND the master-control cells responsible for the formation of a large portion of the sinuses, upper palate, and nasopharynx. When a certain pair of genes is mutated, the result is a failure of recanalization of the nasopharynx either partially or in its entirety, a condition called 'choanal atresia', which is part of the pathology of at least 4 syndromes: Crouzon , Pfeiffer , Antley-Bixler, and Treacher-Collins syndromes.

https://en.wikipedia.org/wiki/Choanal_atresia

The mutations generally causing the entire spectrum of symptoms in these syndromes is known to be mainly due to FGFR2 mutants. And they are not the only syndromes to feature choanal atresia, which may also be non-syndromic. But, how the mutations cause the phenotype isn't understood. I have noted severely abnormal proliferation in cells with a mutation in another cell proliferation pathway, one which usually also triggers cell proliferation when activated (Wnt-Bcat, via receptors called 'Frizzled'). But in this case, it may instead be acting in a regulatory role, co-modulating cell division within the craniofacial region in this specific cell type which are noteworthy because of a transient expression of a stem cell marker called LGR5. It is only active for 2 days during mouse development in this class of cells, but quite critical as completely knocking out the gene results in a lethal phenotype by post-natal day 1. LGR5 is mostly known for its role in intestinal development and colon cancer. Deletions of it have been known to produce the severe phenotype, but exactly how this phenotype arises from the mutation isn't known. I've found that losing LGR5 causes a massive overproliferation of the substype of cell which transiently expresses it (I found them by adding in a constitutive reporter gene rather than relying only on Green Fluorescent Protein in the LGR5-Cre construct, which would disappear in a couple days once the gene shuts off, while the reporter will always be expressed in all cellular descendants once activated by the expression of Cre recombinase from the LGR5 construct in concert with timed tamoxifen administration... it's a rather complicated gene insert) in the developing palate and sinuses, so much so that they end up displacing almost every other cell in the region, which obviously would disrupt normal formation. This is strange since, as I've noted, LGR5's classical role is indirectly upregulating Wnt-Bcat signaling by internalizing two E3 ligases which usually ubiquitinate and deactivate Wnt receptors, thus decreasing Bcat signaling.

So, there are two possibilities: 1. there is an alternate and less-understood Bcat signal which can sometimes counter other cell growth signals (one of those being the FGF pathway, which as its name implies, stimulates the growth of fibroblasts, a major cell type in mesenchymal tissue) and/or 2. The dual pathways are modulating each other and controlling fine aspects of cell growth and migration within the region, a hypothesis bolstered by the existence of a craniofacial-tissue-specific alternatively-spliced isoform of FGFR2 (isoform IIIc) found only in craniofacial mesenchyme... coincidentally, the exact same locations as the cells expressing LGR5. This strongly suggests a relation, and the proof would come from examining the expression of FGFR2IIIc via in situ hybridization. Alas, I won't have the time for this crucial test.

If I can manage to raise about $50K in the next two weeks, it would keep my research going for another month or two, probably long enough for me to perform the crucial tests to demonstrating the merit of my hypothesis and applying either for a full NIH grant for research or putting my research into a program for a PhD at the University of Pennsylvania.

I shall have to see what's required for a Go-Fund Me campaign, and assemble the data I have so far to demonstrate the physical evidence of what I've uncovered so far.