Retinitis Pigmentosa and Its Effect on the Rhodopsin Receptor Name: Vaishnavie Parthipan Student Number: 1068889 Date:

November 20th, 2011

Abstract Retinitis pigmentosa is a disease caused by the mutation of its GPCR, rhodopsin. It results in the deterioration of vision of a prolonged period. Many mutations will lead to the development of the disease. In most cases, mutants prevent the formation of a functional chromophore, which stops the rhodopsin from leaving the endoplasmic reticulum. Accumulation of misfolded opsin results in apoptosis. There is no cure for this disease, but several therapies have improved vision and decreased rate of degeneration in patients. Understanding how the mutant forms and its function will allow for a cure in the future. Introduction G-protein coupled receptors (GPCR) play a large part in controlling the physiological functions in the human body. GPCR have a key role in cell communication. Mutations in the GPCR can cause acquired and inherited diseases. Retinitis pigmentosa is one of many diseases caused by the mutation of GPCR (Schoneberg, Schulz, Biebermann, Hermsdorf, Rompler & Sangkul, 2004). A person suffering with retinitis pigmentosa will often initially experience night blindness (Mendes, Spuy, Chapple & Cheetham, 2005) because rod function is primary affected (Shintani, Schechtman & Gurwood, 2009). Following night blindness, patients often experience progressive loss of peripheral vision and eventually loss of central vision (Mendes, Spuy, Chapple & Cheetham, 2005). As the disease reaches its final stages, the visual field forms a tunnel configuration (Shintani, Schechtman & Gurwood, 2009). Symptoms are first experienced in adolescence. A person with the disease will be completely blind by the time s/he is 50-80 years old (Shintani, Schechtman & Gurwood, 2009). Retinitis pigmentosa is a set of disorders that can be acquired at random or inherited by three different ways. It causes the degeneration of light receptor cells in the retina. The disease can be transmitted as an autosomal dominant trait, autosomal recessive trait or chromosome x linked trait.

Majority of patients with Retinitis pigmentosa carry the autosomal dominant trait, which results in Autosomal Dominant Retinitis Pigmentosa (ADRP). Mutations of rhodopsin, the GPCR associated with retinitis pigmentosa, are the most common cause of ADRP (Dryja, Hahn, Cowley, McGee & Berson, 1991). Signalling Pathway of Rhodopsin Rhodopsin is a visual pigment that is comprised of protein, rod opsin and an internal ligand chromophore. As soon as light strikes the retina, rhodopsin is activated by the isomerisation of 11-cisretinal in the chromophore (Mendes, Spuy, Chapple & Cheetham, 2005). The activation of rhodopsin allows for the binding of membrane-associated proteins (Kalloniatis & Fletcher, 2004). Binding and activation of rhodopsin results in the activation of transducin, which involves the exchange of GDP for GTP. This exchange will cause transducin -subunit-GTP to separate from the transducin (-subunit) complex and rhodopsin. The separated subunit will activate cyclic guanosine monophosphate phosphodiesterase (cGMP) and allow for the hydrolysis of cGMP (Mendes, Spuy, Chapple & Cheetham, 2005). cGMP allows the entry of cation ions in the photoreceptor outer segment (Kalloniatis & Fletcher, 2004). As the levels of cGMP decrease, the cGMP calcium channels close and result in a decrease of calcium influx. Overtime the membrane potential is changes and is transmitted to the brain as a neural signal. Phototransduction is inactivated by several mechanisms that prevent signalling. As a result, rhodopsin is desensitized and mediated by rhodopsin kinase. This prevents rhodopsin from interacting with transducin. Finally, the chromophore is recycled and a new 11-cis-retinal is bind to rhodopsin (Mendes, Spuy, Chapple & Cheetham, 2005). Mutations and Their Effect on the Original Pathway Rhodopsin mutants, as mentioned above, are the cause for most cases of ADRP. There are over one hundred mutations in rhodopsin, which may lead to retinitis pigmentosa. Scientists believe that there

are several classes of ADRP mutations; however only two are major. Class I mutations occur in the C terminus of protein. They resemble wild type rhodopsin and form functional chromophore with 11-cisretinal (Mendes, Spuy, Chapple & Cheetham, 2005) and plasma membrane, but their G protein coupling abilities were altered(Schoneberg, Schulz, Biebermann, Hermsdorf, Rompler & Sangkul, 2004). The rhodopsin in this class were formed normally but were not transported the right way. Class II mutations are the most common in ADRP. The mutations occur in the intradiscal, transmembrane and cytoplasmic domains of rhodopsin. Class II mutants are unable to form functional chromophore with 11-cis-tinal which results in the mutants to be misfolded and remain in the endoplasmic reticulum (ER). The majority of mutants were likely to be Class II mutants (Mendes, Spuy, Chapple & Cheetham, 2005). As mentioned above, Class II mutants are retained in the ER. The ER is responsible for functions such as lipid synthesis and calcium storage. When calcium homeostasis is disturbed, the cell responds by undergoing the unfolded protein response (UPR). Misfolded opsin is increased in the ER, and thus stress in the result can cause the UPR. UPR responds to the ER stress by decreasing the amount of misfolded proteins by slowing protein synthesis and increasing protein degradation. If ER stress is not lessened, the UPR can undergo cell death by apoptosis. The result of apoptosis is the blindness experienced by the patient with retinitis pigmentosa. It is not understood clearly how the mechanisms in place can cause a cell to commit apoptosis; however, it is usually a result of cellular damage, trophic factor withdrawal or external activators (Baumgartner, 2000). Although several mutations cause retinitis pigmentosa, the most common allele known to do so is the P23H. The P23H mutants are present in the intradiscal domain and cause partial and total misfolding which then results in the malfunction of rhodopsin and further lead to cell apoptosis (Garriga, Liu & Khoran, 1996). Another mutation in ADRP is the L125R mutation. This mutation was first found in 1991 and occurs in the transmembrane helix C. The mutations resulted in the changes of Arginine and

Leucine to other amino acids. Studies found that mutant opsin failed to form chromophore in the visible 11-cis-retinal. It was found that in the mutations where Leucine was replaced by Alanine and Phenylalanine, the rhodopsin chromophore tended to form partially. These mutants had a mix of rhodopsin folded the correct way and misfolded opsin (Garriga, Liu & Khoran, 1996). Although there are a large variety of mutations that can cause ADRP, in common they all result in photoreceptor death and functional blindness. Treatment The knowledge of how mutations affect rhodopsin can help in the development of treatment and therapy for this disease. Although there is no cure for this disease at present, many approaches are being used to help patients. Several treatments that targeting the disease include vitamin A pulmitate, gene therapy and retinal transplant. Vitamin A Pulmitate Recent studies suggest nutritional supplements might have the ability to preserve the function of photoreceptors. The supplement used most often in treatment of retinitis pigmentosa is vitamin A palmitate. Patients who take 15000 IU/d of vitamin A palmitate could potentially have a slower development of the disease. It is unsure as to how this helps exactly, however vitamin A plays a role in the formation of rhodopsin, thus it may be likely that may allow it to be beneficial for treatment. (Shintani, Schechtman & Gurwood, 2009). Gene Therapy Gene Therapy is a procedure that allows ones to restore and/or replace the gene causing the disease to restore original gene function. Because retinitis pigmentosa is an inherited disease for the most part, genes can be replaced in a number of ways. To replace a gene, a normal gene can be put in the genome using a carrier vector. Ribozyme therapy and RNA interference can also be used. In ADRP,

the mutant product is harmful and results in apoptosis. Ribozymes are designed and used to cleave mutant mRNA as to prevent the dangerous protein from being produced (Shintani, Schechtman & Gurwood, 2009). Retinal Transplant An artificial silicon retina microchip has proven to be effective in the treatment of retinitis pigmentosa. In a study, the microchip was inserted in the right eye while the left eye served as the control. After a period of 6-18 months, patients were followed up and it was found that no one showed implant rejection, infection, inflammation, retinal detachment or migration. Improvement in visual function, in areas such as brightness, contrast, colour, movement, shape, resolution and visual field size, were seen in all patients (Chow, Chow, Packo, Pollack, Peyman & Schuchard, 2004). Conclusion Retinitis pigmentosa is a disease that causes for the loss of vision. Patients at first experience night blindness; however, over time they feel more symptoms, which leads to complete blindness. Rhodopsin, the GPCR, is a visual pigment that allows for normal vision. As rhodopsin becomes mutated, its mutant forms are unable to produce chromophore, the ligand that is essential for the transport of rhodopsin. Because chromophore is not being produced, mutant stays in the ER. The pathway cannot continue and thus upon the UPR, cells undergo apoptosis. Several therapies have been found that help improve vision, however, no actual cure has been found for the disease. Although no cure is present at this time, increased research into gene therapy would be the most beneficial for this disease. Gene therapy removes the mutated gene; thus, some function of the receptor is restored. If one found a way to replace the gene, it would benefit all those suffering with retinitis pigmentosa, for it would allow for the reconstruction of the pathway that is currently disturbed. As more

information is gained about mutants, it will be easier to find out what causes them and why they function the way they do. This will be the first step in finding a cure for this disease.

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