MicroRNA and metabolomics signatures for adrenomyeloneuropathy disease severity

Abstract

Adrenomyeloneuropathy (AMN), the slow progressive phenotype of adrenoleukodystrophy (ALD), has no clinical plasma biomarker for disease progression. This feasibility study aimed to determine whether metabolomics and micro‐RNA in blood plasma provide a potential source of biomarkers for AMN disease severity. Metabolomics and RNA‐seq were performed on AMN and healthy human blood plasma. Biomarker discovery and pathway analyses were performed using clustering, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and regression against patient's clinical Expanded Disability Status Score (EDSS). Fourteen AMN and six healthy control samples were analyzed. AMN showed strong disease‐severity‐specific metabolic and miRNA clustering signatures. Strong, significant clinical correlations were shown for 7‐alpha‐hydroxy‐3‐oxo‐4‐cholestenoate (7‐HOCA) (r2 = 0.83, p < 0.00001), dehydroepiandrosterone sulfate (DHEA‐S; r2 = 0.82, p < 0.00001), hypoxanthine (r2 = 0.82, p < 0.00001), as well as miRNA‐432‐5p (r2 = 0.68, p < 0.00001). KEGG pathway comparison of mild versus severe disease identified affected downstream systems: GAREM, IGF‐1, CALCRL, SMAD2&3, glutathione peroxidase, LDH, and NOS. This feasibility study demonstrates that miRNA and metabolomics are a source of potential plasma biomarkers for disease severity in AMN, providing both a disease signature and individual markers with strong clinical correlations. Network analyses of affected systems implicate differentially altered vascular, inflammatory, and oxidative stress pathways, suggesting disease‐severity‐specific mechanisms as a function of disease severity.

iPSC-derived astrocytes to model phenotype-specific differential neuroinflammatory and metabolic responses in X-linked adrenoleukodystrophy

“…IGF-dysfunction is reported in ALD fibroblasts (Al-Essa and Dhaunsi, 2019) and insulin signaling in reduced in ABCD1-KO mice spinal cord (Schluter A et al, 2012). Interestingly, we recently documented dysregulated IGF signaling as one of the hubs in plasma metabolite pathways associated with AMN disease severity (Turk BR, 2022). Additionally, we observed upregulated miR-409-3p in AMN and cALD astrocytes.…”

“…21 Interestingly, we recently documented decreased miR-409-3p expression in plasma of AMN patients with increasing disease severity. 47 Similarly, our differential gene IPA network shows that miR-145-3p is associated with HAND1, ATRX, and RAP2C genes. Altered ATRX and RAP2C have been reported to be associated with occurrence of gliomas.…”

“…Reports have shown that miR-409-3p contributes to IL-17-induced inflammatory cytokine production in astrocytes via STAT3 signaling pathway in experimental autoimmune encephalomyelitis (EAE) mice (Liu et al, 2019). miR-409-3p expression was also decreased in the plasma of patients with AMN with increasing disease severity (Turk BR, 2022). Similarly, comparative IPA network analysis of CTL vs AMN showed that miR-145-3p is associated with HAND1, ATRX, and RAP2C genes.…”

“…Inspired by our recent identification of plasma biomarker of disease severity in AMN phenotype (Turk BR, 2022) and to identify upstream regulators of inflammatory pathways, we performed comprehensive miRNA profiling in CTL, AMN and cALD iPSC-derived astrocytes and revealed previously unidentified differentially expressed miRNAs in AMN and cALD astrocytes. Most interestingly, we observed a disease-severity associated increase in miR-9-3p and miR-9-5p expression (CTL<AMN<cALD).…”

“…Mature miRNA sequences were mapped to the precursor miRNA sequences of miRBase v22.1 (Bowtie 1.1.2). (Langmead and Salzberg, 2012) miRNA analysis was performed using miRDeep2 (miRDeep2 2.0.0.8) (Friedlander et al, 2012) and R package DEseq2 as published earlier (Turk BR, 2022). Boxplots of miRNA abundance (log10) within the three groups were drawn for altered miRNAs.…”