Demographic and other key characteristics of the CHOP patient cohort (N = 30).

Demographic and other key characteristics of the CHOP patient cohort (N = 30).</p

Demographic and other key characteristics of the RDCRN cohort (N = 290).

Demographic and other key characteristics of the RDCRN cohort (N = 290).</p

RDCRN mitochondrial disease subject validation cohort.

<p><b>A. Experienced Symptoms.</b> Frequency of experienced symptoms as reported by the RDCRN self-reported cohort revealed muscle weakness, chronic fatigue, exercise intolerance, imbalance, and gastrointestinal problems to be the top 5 common symptoms (n = 290). <b>B. Prioritized Symptoms for trial participation.</b> The RDCRN cohort reported the same top 5 symptoms that would motivate trial participation (n = 290) as seen in the CHOP cohort (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0197513#pone.0197513.g001" target="_blank">Fig 1</a>). <b>C. Likely to participate if experienced symptom is targeted in a clinical trial.</b> The RDCRN cohort reported muscle weakness, chronic fatigue, exercise intolerance, diabetes and kidney disease as the most common experienced and desirable to treat symptoms in a clinical trial (n = 290).</p

CHOP mitochondrial disease subject discovery cohort.

<p><b>A. Experienced Symptoms.</b> The CHOP PMD subject cohort reported muscle weakness, chronic fatigue, exercise intolerance, imbalance, and gastrointestinal problems as the top 5 most commonly experienced symptoms (n = 30). <b>B. Prioritized Symptoms for trial participation.</b> The CHOP PMD patient cohort reported the top 5 most commonly experienced symptoms as the same leading symptoms to motivate their trial participation (n = 30). <b>C. Likely to participate if experienced symptom is targeted in a clinical trial.</b> All CHOP PMD subjects who experienced muscle weakness (n = 37), peripheral neuropathy (n = 13), tinnitus (n = 7), diabetes (n = 4), and stroke (n = 3) reported they would participate in a clinical trial that targeted these symptoms.</p

A novel <i>HSD17B10</i> mutation impairing the activities of the mitochondrial RNase P complex causes X-linked intractable epilepsy and neurodevelopmental regression

<p>We report a Caucasian boy with intractable epilepsy and global developmental delay. Whole-exome sequencing identified the likely genetic etiology as a novel p.K212E mutation in the X-linked gene <i>HSD17B10</i> for mitochondrial short-chain dehydrogenase/reductase SDR5C1. Mutations in <i>HSD17B10</i> cause the HSD10 disease, traditionally classified as a metabolic disorder due to the role of SDR5C1 in fatty and amino acid metabolism. However, SDR5C1 is also an essential subunit of human mitochondrial RNase P, the enzyme responsible for 5′-processing and methylation of purine-9 of mitochondrial tRNAs. Here we show that the p.K212E mutation impairs the SDR5C1-dependent mitochondrial RNase P activities, and suggest that the pathogenicity of p.K212E is due to a general mitochondrial dysfunction caused by reduction in SDR5C1-dependent maturation of mitochondrial tRNAs.</p

Generation of a <i>Pdss2</i> Conditional Null Allele.

<p>Generation of a <i>Pdss2</i> conditional null allele, showing a map of the <i>Pdss2</i> genomic locus and the targeting vector with exons represented by open boxes. The relative position of PCR primers (small arrowheads), loxP (large arrowheads), as well as cassettes encoding neomycin phosphotransferase (neo) are shown. Primers koF, koF2 and koR were used in PCR genotype analysis. Cre-mediated deletion results in either the <i>Pdss2</i> null allele (deletion of exon 3 ) or the <i>Pdss2 ^loxP/loxP</i> allele (exon 2 flanked by loxP sites). Abbreviations: Ba, BamH I; Aa. Aat II; Xh, Xho I; Dr, Dra III.</p

Metabolic Pathway Alterations Are Seen by Expression Profiling in B6.<i>Alb/cre,Pdss2^loxP/loxP</i> Mouse Liver.

<p>Global genome expression profiling in B6.<i>Alb/cre,Pdss2^loxP/loxP</i> mouse liver identifies concordant transcriptional alterations interpretable at the level of multiple metabolic pathways, which suggest significantly altered intermediary metabolism occurs despite an apparent absence of symptomatic disease. Extensive evolutionary concordance in upregulation of key biochemical pathways is seen in primary mitochondrial dysfunction, both in this mammalian <i>Pdss2</i> liver-conditional knockout model of coenzyme Q deficiency and in a previously reported <i>C. elegans gas-1(fc21)</i> missense mutant model of primary complex I dysfunction <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1000061#pgen.1000061-Falk1" target="_blank">[21]</a>. Biochemical pathways as curated from the KEGG online database (<a href="http://genome.jp.kegg" target="_blank">http://genome.jp.kegg</a>) are indicated with the # of genes in each pathway (cluster size), normalized enrichment score (NES), statistical significance of altered pathway expression between mutant and wildtype controls (<i>p</i>-value), and false positive percentage in the form of a false discovery rate (FDR <i>q</i>-value) according to GSEA. Pathways are ranked by descending NES in the <i>Pdss2</i> mutant (left data column). Comparison to previously reported complex I <i>gas-1</i>(<i>fc21</i>) missense <i>C. elegans</i> mutant dataset (middle data column) and a “validation” <i>C. elegans</i> dataset of 8 different complex I, II, and III missense and RNAi-interference generated mutants (right data column) is indicated by differential highlights <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1000061#pgen.1000061-Falk1" target="_blank">[21]</a>. Font color denotes a pathway as relatively upregulated (red), downregulated (green), or unchanged (black).</p

Histologic Features of Renal Disease in <i>Pdss2</i> Mutant and Control Mice.

<p>Histologic features of renal disease in mutant and control mice (H & E-stained sections, original magnifications all 200x). <i>A</i>, B6.<i>Podocin/cre,Pdss2^loxP/loxP</i> mouse (290 days old; 64 mg albumin; histologic score, 4+). <i>B,</i> B6.<i>PEPCK/cre</i>, <i>Pdss2^loxP/loxP</i> (191 days old; 0.04 mg albumin; histologic score 0). <i>C</i>, B6.<i>Pdss2^kd/kd</i> (146 days old; 15 mg albumin; histologic score, 3+). <i>D</i>, B6. <i>Pdss2^loxP/loxP</i> mouse, (191 days old; 0.12 mg albumin; histologic score, 0). Note the prominent tubular dilatation and interstitial infiltrates in panels A and C, but absent in panels B and D.</p

<i>Pdss2</i> Conditional Knockout Confirmation.

<p>PCR products after laser capture microdissection, using primers for exon 2 (A) or exon 4 (B). Lanes: 1, <i>Pdss2^loxP/loxP</i> glomerulus; 2, <i>Pdss2^loxP/loxP</i> tubules; 3, B6.<i>Podocin/cre,Pdss2^loxP/loxP</i> mouse 1, glomerulus; 4, B6.<i>Podocin/cre,Pdss2^loxP/loxP</i> mouse 1 tubules; 5, B6.<i>Podocin/cre,Pdss2^loxP/loxP</i> mouse 2, glomerulus; 6, B6.<i>Podocin/cre,Pdss2^loxP/loxP</i> mouse 2 tubules; 7, B6.<i>PEPCK/cre</i>,<i>Pdss2^loxP/loxP</i> mouse 1 glomerulus; 8, B6.<i>PEPCK/cre,Pdss2^loxP/loxP</i> mouse 1, tubules; 9, B6.<i>PEPCK/cre</i>,<i>Pdss2^loxP/loxP</i> mouse 2 glomerulus; 10, B6.<i>PEPCK/cre,Pdss2^loxP/loxP</i> mouse 2, tubules.</p

Glomerular Electron Micrographs From <i>Pdss2</i> Mutant and Control Mice.

<p>Electron micrographs from mutant and control mouse kidney glomeruli; original magnifications all 10,000x; scale bar = 2 microns. <i>A</i>, B6, 362 days old; arrows show podocyte foot processes. <i>B,</i> B6.<i>Pdss2^kd/kd</i>, 267 days old; arrows show regions of foot process effacement. <i>C</i>, B6.<i>Pdss2^loxP/loxP</i> mouse, 248 days old; arrows show foot processes. <i>D</i>, B6.<i>Podocin/cre,Pdss2^loxP/loxP</i> 290 days old; arrows show regions of foot process effacement.</p