CLN2/TPP1 deficiency: the novel mutation IVS7-10A>G causes intron retention and is associated with a mild disease phenotype.

The classical form of late infantile neuronal ceroid lipofuscinosis (LINCL) is a childhood hereditary neurodegenerative disease usually fatal in the first decade of life. The underlying gene, CLN2, encodes the lysosomal soluble enzyme tripeptidyl-peptidase 1 (TPP1). In a Portuguese patient with juvenile form of the disease, the histochemical study revealed the presence of curvilinear inclusions typical of LINCL. In vitro TPP1 activity was deficient in patient's cells. CLN2 gene analysis revealed the transition IVS7-10A>G (g.4196A>G) in both alleles. In silico analysis suggested that A-to-G change in the A-rich region of intron 7 could cause aberrant splicing of exon 8 by creating a novel acceptor splice site. However, because the wild-type acceptor of intron 7 is weak and it was not apparently affected, the severity of this mutation could not be established through sequencing data of gDNA. Normal level of spliced CLN2/mRNA was observed in patient's fibroblasts. In the cDNA, the 9-nt retention of intronic sequence (c.886_887ins9) was observed. The mutation is predicted to result in a protein with three extra amino acids between proline 295 and glycine 296. In patient's fibroblasts the level of mutant CLN2p was reduced to about 60% but the migration pattern was similar to the wild-type protein, suggesting that it was correctly targeted to the lysosomes. Taken together, these findings suggest that the first "ag" is selected for splicing and the mutant protein must retain some residual catalytic activity, thus explaining the late onset and the delayed progression of the disease.