Royal Society of Edinburgh Henry Dryerre Prize Lecture Making Eyes Lessons from Failed Miracles Professor Veronica

a van Heyningen University of Edinburgh Institute of Genetics and Molecular Medicine/ MRC Human Genetics Unit 7 September 2009 Report by Jennifer Trueland
Charles Darwin described the eye as the organ of extreme perfection and even wondered whether the principles of evolution would hold true where it was concerned. In a lecture described as a tour de force, Veronica van Heyningen considered the genetics of the eye and the light that study of the subject casts on diseases both rare and common. The amazing thing about development is not so much that it sometimes goes wrong, but that it ever goes right, said Professor van Heyningen. The processes involved in building the eye, for example, are so complex, that there are a near-infinite number of possibilities for it going wrong. Mutations can help us understand normal biology. Looking at where development has gone wrong is important, because it opens a window to the mechanisms of disease. This information can help lead to drug discovery, or, as is more likely in the case of serious eye disorders, can improve disease management. The very development of the eye, intimately bound as it is with brain development, is so complex, that it is not surprising that it sometimes goes wrong, often as the result of altered gene function. Professor van Heyningen spoke about three major genes with key roles in eye development and function: PAX6, SOX2 and OTX2. Each is involved in development of the eye, the brain (and other parts of the body) and each is implicated in one or more serious eye malformations. All work as DNA-binding transcription factors (TFs), which regulate expression of other genes, including those in the eye. All three genes are dosage-sensitive in humans: losing the function of one copy, through mutation, generally causes serious eye malformations. Studying this, and how it comes about, has given important information about how normal development and maintenance happens, as well as how the disease occurs. Using animal models, it has also been possible to explore the role of environmental factors in the severity of the malformation. Visual impairment is relatively common, she said, partly because we, in human society, look after people with eye problems well, and they can survive and have children. Even before the Human Genome Project, there was a great deal of work looking at inherited eye disease, trying to find the genes responsible.

Professor van Heyningen and her colleagues have looked at a number of eye conditions associated with chromosomal disorder. These include aniridia (where the iris is missing). The van Heyningen group was involved in proving that PAX6 mutations cause the abnormality. The other transcription factors, SOX2 and OTX2, were later found to be implicated in a proportion of rare anophthalmia (no eye globe) and microphthalmia (small eye). Each of these genes is involved in eye development at several different stages and, importantly, all are highly conserved; that is, they are found in even very primitive forms of life. This is particularly the case with PAX6, where mutations affecting the sensory system can be found even in a nematode worm which has no eyes. The genes are also found in mouse models, and in zebrafish and the fruit fly, each of which has different useful features as a research model. As well as animal studies, however, research into people with aniridia, using MRI scanning, has shown abnormalities in the brain. A high proportion of the patients, for example, are missing a working version of one of the key connections between the brains two hemispheres, while many also had problems with the smell system. Problems with absence of the pineal (important for sleep) were also a surprise finding. Later, hearing problems were identified because information could not transfer efficiently from the left to the right ear. The research has shown that the genes work in different ways in different people, with the effects varying even within families sometimes there are parents who are carriers of the mutant gene but who show no effects, for example. Severity of condition can also vary between brothers and sisters. The genes themselves are embedded within a complex array of regulatory elements and there have been cases where aniridia, for example, appears to have been caused by chromosomal breakpoints outside the gene. This suggests that changes in gene regulation are also playing an important role, so the genes themselves cannot be taken in isolation. The genes are subject to various enhancers which affect how they work. How the gene behaves also seems to depend on the stage of development and it is now considered that changes in gene regulation are a major mechanism for evolutionary change. Regulatory variants may be the reason why some of us are predisposed to develop later-onset common eye diseases (such as cataracts and glaucoma). Two or more transcription factors can be at work in the same developmental pathways another example of the complex dance which is being played out in time and space. Bioinformatics using mathematics to better predict gene targets is also proving useful, using a method called Hidden Markov Modelling. The computer findings are being tested out in zebrafish to confirm their accuracy. Environmental factors may also be at play in determining whether someone develops an abnormality. For example, work on the fruit fly has shown that the effect of existing mutations were only revealed when the heat shock protein 90 (HSP90) was disturbed. The Edinburgh team has shown that the same thing happens in zebrafish, where it is possible to uncover hidden mutations as the cause of disease by manipulating HSP90. These findings raised more questions, about proteins which interact with HSP90. Excitingly, this has shown that HSP90 protein interacts with a partner protein that can be found in the primary cilia and ciliary abnormalities are found in many diseases, such as diabetes and heart defects. Again, this is an example where studying mutations in rare diseases may have important implications for common diseases.

In conclusion, looking at rare abnormalities gives valuable clues about the complex interplay of proteins, genes and other factors which go into the development of both normal and abnormal organs. Work on genes known to cause rare eye conditions may also lead to important discoveries for understanding the common diseases that are a major cause of death and morbidity across the world. Exploring these highly conserved genes along with environmental factors may provide a real route to drug treatments and better management of many diseases, not just those of the eye. Questions Questions ranged from genetics to evolutionary history, via salmon. Asked what she meant by describing a mutant form of PAX6 (in terms, for example, of where it was mutated) Professor van Heyningen said that in most cases mutations led to loss of function, so that no protein was made from the gene. In other cases, there was an amino acid change in the protein, usually in the DNA-binding paired domain which altered protein behaviour. There could also be mutations in other parts of the gene, but one of the challenges is knowing which phenotypes were involved and therefore where to look for such mutations. A recent suggestion that a PAX6 regulatory region mutation was involved in a particular kind of childhood epilepsy was worth looking at, she said. A scientist from the University of Idaho described how experiments involving salmon showed that it was possible to reduce the number born with mutations if they were treated with kid gloves. Professor van Heyningen said that this backed the idea that heat shock or another insult during development could cause a problem where there was an underlying genetic predisposition the idea that environment, as well as genetics, has a role to play. Asked by a retired physician what her view was about the Scottish Governments emphasis on funding research into common diseases, Professor van Heyningen said that research into rarer conditions could provide valuable information about the mechanisms of disease, which help us learn more about common diseases. Finally, a member of audience asked at what point in the evolution of life-forms the ability to see began. If the earliest forms of life were blind, how long did it take for sight to develop, and would we always have it? Professor van Heyningen said she wasnt an evolutionary biologist, but pointed out that many early organisms could sense light even without eyes. Some of the earliest creatures including some nematodes and jellyfish, had forms of eyes, and even some bacteria react to light. Vote of Thanks The vote of thanks was delivered by Professor Nick Hastie, director of the MRC Human Genetics Unit and of the Institute of Genetics and Molecular Medicine at the University of Edinburgh. He called the lecture a tour de force and said that Professor van Heyningen was a world-leading researcher, who had seen the potential of human genetics to understand biology ahead of everyone else.

Opinions expressed here do not necessarily represent the views of the RSE, nor of its Fellows The Royal Society of Edinburgh, Scotlands National Academy, is Scottish Charity No. SC000470