A new diagnostic indication device of a biomarker growth differentiation factor 15 for mitochondrial diseases: From laboratory to automated inspection

Abstract

Mitochondrial diseases (MDs) are occasionally difficult to diagnose. Growth differentiation factor 15 (GDF15) has been reported as a biomarker useful for not only diagnosing MDs, but also evaluating disease severity and therapeutic efficacy. To enable the measurement of serum GDF15 concentrations at medical institutions, we developed a new latex‐enhanced turbidimetric immunoassay (LTIA) as an automated diagnostic indication test for MDs. We also examined the equivalency of specificity and sensitivity in measuring serum GDF15 concentrations between a commercially available enzyme‐linked immunosorbent assay (ELISA) kit and a novel LTIA device in patients with MDs, disease controls, and healthy controls. A clinical performance study used a newly developed LTIA device and an existing ELISA kit to measure the concentrations of GDF15 in 35 MD patients, 111 disease controls, and 86 healthy controls. The median (first quartile‐third quartile) of serum GDF15 concentrations measured with the LTIA device was significantly higher (P < .001) in MD patients (1389.0 U/mL [869.5‐1776.0 U/mL]) than in healthy controls (380.5 U/mL [330.2‐471.8 U/mL]); the interquartile ranges did not overlap between MD patients and healthy controls. The areas under the curve in disease and healthy controls were 0.812 (95% confidence interval [CI]: 0.734‐0.886) and 0.951 (95% CI: 0.910‐0.992), respectively. The automated, high‐throughput technology‐based LTIA device has definite advantages over the ELISA kit in shorter processing time and lower estimated cost per sample measurement. The LTIA device of GDF15 may be a sufficiently reliable, frontline, diagnostic indicator of individuals with suspected MDs in the general population.

Synthetic aporphine alkaloids are potential therapeutics for Leigh syndrome

“…GDF-15 is a biomarker that is elevated in the blood of patients with mitochondrial diseases and is used for the diagnosis of mitochondrial diseases and determination of therapeutic efficacy 26 , 36 . It has also been reported to be upregulated in multiple sclerosis, cardiac failure, and liver disease 37 – 39 .…”

Neuropathological hallmarks in autopsied cases with mitochondrial diseases caused by the mitochondrial 3243A>G mutation

“…Immunohistochemical examinations were performed as previously described [ 11 ]. The sections were incubated overnight with the following primary antibodies: Human Mitochondria antibody (M117, cline AF‐1; Leinco Technologies Inc., MO, USA; 1:200, pretreated by heat antigen retrieval), human PHF‐Tau (monoclonal, clone AT8; Thermo Scientific, IL, USA; 1:5000), amyloid β [ 4 , 5 , 6 , 9 , 10 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 ] (monoclonal; Dako, Glostrup, Denmark; 1:1000, pretreated by heat antigen retrieval and formic acid), α‐synuclein (polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:20,000, pretreated by heat antigen retrieval and formic acid), and phosphorylated trans‐activation response DNA‐binding protein of 43 kDa (p‐TDP‐43, polyclonal; CosmoBio, Tokyo, Japan; 1:4000, pretreated by heat antigen retrieval and formic acid). Then, these sections were washed with phosphate‐buffered saline (PBS) and incubated with a secondary antibody (Histofine Simple Stain MAX PO (MULTI); Nichirei Bioscience Inc., Tokyo, Japan) for 1 h. The sections were visualized using 3,3′‐diaminobenzidine (DAB Tablet; FUJIFILM, Osaka, Japan), and Mayer's hematoxylin solution was used as a counterstain.…”

“…1.53 t software ( https://imagej.nih.gov/ij/ ) and HE images taken with a ×100 objective lens (Figure 1L, M, P, Q ). Based on each diagnostic criteria, Alzheimer's disease was assessed using PHF‐Tau and amyloid β [ 4 , 5 , 6 , 9 , 10 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 ] IHC of the temporal lobe [ 20 , 25 ], Lewy body disease was assessed using α‐synuclein IHC of the substantia nigra, locus coeruleus, and dorsal nucleus of the vagus nerve [ 19 ], cerebral amyloid angiopathy was assessed using amyloid β [ 4 , 5 , 6 , 9 , 10 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 ] IHC of the temporal and occipital lobes [ 18 ], and argyrophilic grains was assessed using PHF‐Tau IHC and Gallyas‐Braak staining of the amygdala and medial temporal lobe [ 26 ].…”

“…A study using a cytoplasmic hybrid technique revealed that >90% mtDNA heteroplasmy is needed to decrease oxidative phosphorylation (OXPHOS) [ 5 ], which is responsible for the bulk of cellular ATP. Recently, it was reported that serum fibroblast‐growth factor 21 (FGF21) [ 6 , 7 , 8 ] and growth and differentiation factor 15 (GDF15) [ 9 , 10 ] are elevated in patients with mitochondrial disease, but the roles of FGF21 and GDF15 in the brain tissue remain unclear. Thus, we investigated the mtDNA heteroplasmy, mitochondrial content, and OXPHOS complexes using brain tissue samples in the autopsied mutant cases and compared the results to their neuropathological findings.…”

Editorial: Mitochondrial medicine special issue

“…This issue of JIMD provides a sampling of the gamut of presentations at this meeting, from fundamental research 1 to addressing practical hurdles in drug development, including how to work with regulatory agencies, 2 what is required by industry to move from an early concept to a licensed medicine, 3 and the critical need to develop reliable noninvasive biomarkers of disease progression 4 and other outcome measures to allow effective evaluation of new compounds 5 . Recent developments in animal models were presented, 6 including high throughput approaches for drug screening 7 .…”