@article {957,
	title = {Multiomic analysis elucidates Complex I deficiency caused by a deep intronic variant in NDUFB10.},
	journal = {Hum Mutat},
	volume = {42},
	year = {2021},
	month = {2021 Jan},
	pages = {19-24},
	abstract = {<p>The diagnosis of Mendelian disorders following uninformative exome and genome sequencing remains a challenging and often unmet need. Following uninformative exome and genome sequencing of a family quartet including two siblings with suspected mitochondrial disorder, RNA sequencing (RNAseq) was pursued in one sibling. Long-read amplicon sequencing was used to determine and quantify transcript structure. Immunoblotting studies and quantitative proteomics were performed to demonstrate functional impact. Differential expression analysis of RNAseq data identified significantly decreased expression of the mitochondrial OXPHOS Complex I subunit NDUFB10 associated with a cryptic exon in intron 1 of NDUFB10, that included an in-frame stop codon. The cryptic exon contained a rare intronic variant that was homozygous in both affected siblings. Immunoblot and quantitative proteomic analysis of fibroblasts revealed decreased abundance of Complex I subunits, providing evidence of isolated Complex I deficiency. Through multiomic analysis we present data implicating a deep intronic variant in NDUFB10 as the cause of mitochondrial disease in two individuals, providing further support of the gene-disease association. This study highlights the importance of transcriptomic and proteomic analyses as complementary diagnostic tools in patients undergoing genome-wide diagnostic evaluation.</p>},
	issn = {1098-1004},
	doi = {10.1002/humu.24135},
	author = {Helman, Guy and Compton, Alison G and Hock, Daniella H and Walkiewicz, Marzena and Brett, Gemma R and Pais, Lynn and Tan, Tiong Y and De Paoli-Iseppi, Ricardo and Clark, Michael B and Christodoulou, John and White, Susan M and Thorburn, David R and Stroud, David A and Stark, Zornitza and Simons, Cas}
}
@article {709,
	title = {Clinical Spectrum and Functional Consequences Associated with Bi-Allelic Pathogenic  Variants.},
	journal = {J Clin Med},
	volume = {8},
	year = {2019},
	month = {2019 Nov 19},
	abstract = {<p> (PNPase-polynucleotide phosphorylase) is involved in multiple RNA processing functions in the mitochondria. Bi-allelic pathogenic  variants cause heterogeneous clinical phenotypes affecting multiple organs without any established genotype-phenotype correlations. Defects in PNPase can cause variable combined respiratory chain complex defects. Recently, it has been suggested that PNPase can lead to activation of an innate immune response. To better understand the clinical and molecular spectrum of patients with bi-allelic  variants, we captured detailed clinical and molecular phenotypes of all 17 patients reported in the literature, plus seven new patients, including a 78-year-old male with the longest reported survival. A functional follow-up of genomic sequencing by cDNA studies confirmed a splicing defect in a novel, apparently synonymous, variant. Patient fibroblasts showed an accumulation of mitochondrial unprocessed  transcripts, while blood showed an increased interferon response. Our findings suggest that functional analyses of the RNA processing function of PNPase are more sensitive than testing downstream defects in oxidative phosphorylation (OXPHPOS) enzyme activities. This research extends our knowledge of the clinical and functional consequences of bi-allelic pathogenic  variants that may guide management and further efforts into understanding the pathophysiological mechanisms for therapeutic development.</p>},
	issn = {2077-0383},
	doi = {10.3390/jcm8112020},
	author = {Rius, Rocio and Van Bergen, Nicole J and Compton, Alison G and Riley, Lisa G and Kava, Maina P and Balasubramaniam, Shanti and Amor, David J and Fanjul-Fernandez, Miriam and Cowley, Mark J and Fahey, Michael C and Koenig, Mary K and Enns, Gregory M and Sadedin, Simon and Wilson, Meredith J and Tan, Tiong Y and Thorburn, David R and Christodoulou, John}
}
