case_images: PMC10613489_01
case_images: Cranial MRI of the patient. Axial T1 and T2-weighted imaging showed hypointense and hyperintense signals in the thalamus, mesocephalon, and pons (indicated in red arrow).
case_images: fneur-14-1270793-g0001.jpg
case_images: PMC10613489_01_fneur-14-1270793-g0001.jpg
case_images: F1
case_images: T2 hyperintense signals in the thalamus, mesocephalon, and pons without gadolinium enhancement (see Figure 1). The cerebrospinal fluid (CSF) showed no abnormalities.
case_images: (A) Pedigree chart of this family. (B) Validation of SCP2 c.674 + 1 G>C by Sanger sequence in the patient and his father. The ref indicates the reference sequence, with capital letters and lowercase, representing the nucleotide in the exon and intron regions. (C) Outlier-level significance [-log10(p), y-axis] vs. effect size (junction count/total junction count-mean (total junctions count/total count of junctions in the cluster), x-axis) for the patient. There are 2030s outliers with an adjusted p < 0.05 in 1442 genes (red dots indicating two clusters in SCP2, vertical dotted lines indicating the effect size cutoff). (D) Sashimi plot of the eighth exon-skipping event (indicated in black arrow) in RNA-seq samples of the SCP2-affected (red) and three representative SCP2-unaffected (orange) individuals. The RNA-seq read coverage is given as the log10 RPKM-value (Reads Per Kilobase of transcript per Million mapped reads, y-axis), and the number of splits reads spanning an intron is indicated on the exon-connecting line. (E) The skipping event is observable in the RT-PCR product from patient blood. Mt: mtant type, wt: wide type. (F) Exon 8 skipping is validated by the Sanger sequencing of RT-PCR product. (G) schematic representation of the SCPx and the affected domain of the index patient, surrounding the amino acids region coding by the eighth exon (p. 225-254, indicated between the red cross). (H) Protein sequence alignment of SCP2 orthologs, showing the region surrounding the p. 225-254 is relatively conserved (indicated between the red dot lines). *Indicates the termination codon.
case_images: fneur-14-1270793-g0002.jpg
case_images: PMC10613489_01_fneur-14-1270793-g0002.jpg
case_images: F2
case_images: The proband's father and sisters were heterozygotes at this position (see Figures 2A, B). RNA sequencing identified two outlier junctions in the SCP2 cluster, providing support for the skipping of the eighth exon (p = 4.81E-12, FDR = 0, and p = 1.31E-4, FDR = 0.008) (see Figures 2C, D). At the mRNA level, cDNA analysis validated the aberrant splicing pattern between the sixth and ninth exons in the patient's sample through the agarose gel electrophoresis of reverse transcription polymerase chain reaction amplicons (RT-PCR) product (see Figure 2E). Sanger sequencing of the RT-PCR product confirmed the exon 8 skipping event in the patient (see Figure 2F).