Using cGMP analogues to modulate photoreceptor light sensitivity.

Abstract

For many hereditary degenerative diseases, like retinitis pigmentosa, the primary degeneration of rod photoreceptors leads to a secondary loss of cones and eventually to blindness. The pathophysiological mechanism is very often characterized by an increase of cGMP level in rods. Thus, rod degeneration is mainly triggered by an over-activation of cyclic nucleotide-gated (CNG) channels and excessive Ca2+ influx. While targeting of CNG channels was already proposed for therapeutic purposes, thus far, it has not been possible to selectively inhibit rod CNG channels without compromising cone function. Here, we present a novel strategy, based on a combination of cGMP analogues, which enables the selective modulation of rod photoreceptor activity. This approach was first tested by means of the patch-clamp technique on retinal CNG channels, heterologously expressed in Xenopus oocytes and further confirmed by micro-electroretinography recordings on wild-type mouse retina. The cGMP-analogues mixture contained a CNG-channel inhibitor (Rp-8-Br-PET-cGMPS) and a CNG-channel activator (8-pCPT-cGMP). While Rp-8-Br-PET-cGMPS, when applied alone, behaved as a partial agonist and inhibited both cone and rod CNG channels equally, 8-pCPT-cGMP behaved as a very potent agonist and showed a strong concentration-dependent cone selectivity. Co-application of both compounds inhibited rod CNG channels and preserved cone functionality under both physiological and pathological cGMP levels. While the kinetics of rod channels was not influenced by this treatment, the deactivation of cone channels was slowed down. Surprisingly, this aspect did not affect the physiological cone responsiveness to light, indicating possible intracellular compensatory mechanisms. In conclusion, by delaying primary rods degeneration, this strategy shields the healthy cones and thus may either postpone the onset or slow down the disease development. Beyond the treatment of retinal diseases, using cGMP analogues with desired properties may elegantly address the isoform-specificity problem in future pharmacological therapies of related diseases.