Research

Research Methods

In collaboration with Wojciech Kedzierski, Ph.D., we have generated transgenic mouse lines expressing chimeric (part mouse/part human) Cfh molecules with either the 402-histidine (“at-risk”) variant or the 402-tyrosine (“non-risk”) variant of Cfh.

We aim to use these mice to: (a) identify the molecular basis for both the increased susceptibility to AMD and the resistance to therapy observed in human H-CFH carriers, and to (b) generate a relevant model of AMD that would allow the dissection of the role of the different arms of the immune system in the disease.

We have shown that our mice express the transgenic mRNA both in the liver and in the posterior segment ocular tissues. They also produce the transgenic protein, which can be found in the serum.

transgenic mice
Figure 2: Constructs and design of transgenic animals
 
graph
Figure 3: Fold increase in mRNA level by RT-PCR. The posterior segment of 2 transgenic mouse lines vs. non-transgenic mice were analyzed using primers specific for the transgenic mRNA
 
western blot
Figure 4: Western blot and serum concentration of transgenic Cfh in some of our lines. Lines are 4th generation (313 is 2nd gen).
 Fluorescein angiogram
Figure 5: Fluorescein angiogram of laser-induced CNV in a B6 mouse.

We are also using a laser-induced model of AMD as a way to study the acute neovascular phase of the disease. When applied to our transgenic mice it will help us determine if and how the factor H variants affect the susceptibility to neovascular changes, and the dependence of the neovascular membranes on VEGF and other growth factors.

This laser model is also helping us to examine the potential therapeutic role of some antibodies and small molecules generated here at UTSW in blocking or causing regression of choroidal neovascularization.