Genetics: FBXL4

[Hutan]

DecodeME candidate ME/CFS gene

In DecodeME, we attempted to link GWAS variants to target genes. Here we discuss the top two tiers of predicted linked genes that we are most confident about –‘Tier 1’ and ’Tier 2’.

We defined genes as Tier 1 genes if: (i) they are protein-coding genes, (ii) they have GTEx-v10 expression quantitative trait loci (eQTLs) lying within one of the FUMA-defined ME/CFS-associated intervals, and (iii) their expression and ME/CFS risk are predicted to share a single causal variant with a posterior probability for colocalisation (H4) of at least 75%. For this definition, we disregarded the histone genes in the chr6p22.2 HIST1 cluster, as their sequences and functions are highly redundant (1). This prioritisation step yielded 29 Tier 1 genes.

For the intervals without Tier 1 genes, three Tier 2 genes were defined as the closest protein-coding genes without eQTL association: FBXL4 (chr6q16.1), OLFM4 (chr13q14.3), and CCPG1 (chr15q21.3).

CHROMOSOME 6q

Chr6q contained no Tier 1 genes, but one Tier 2 gene.

The interval also contains a non-protein long noncoding RNA locus (RP11-436D23.1) (25) which contains a miRNA locus (miR-2113) of unknown function. The allele that increases the risk of ME/CFS is associated with increasing RP11-436D23.1 expression in four tissues: amygdala, anterior cingulate cortex, cortex and hippocampus.

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FBXL4 (Tier 2)

• Protein: F-box/LRR-repeat protein 4. UniProt. GeneCards.

• Molecular function: Component of the mitochondria-localised SCF-FBXL4 ubiquitin E3 ligase complex. This complex restricts mitophagy by controlling the degradation of BNIP3 and NIX mitophagy receptors (26,27).

• Cellular function: Regulator of mitophagy.

• Link to disease: Mutations in FBXL4 can cause mitochondrial DNA depletion syndrome caused by elevated mitophagy (28).

• Potential relevance to ME/CFS: Reduction in FBXL4 function is associated with impaired mitochondrial respiratory chain deficiency, which has been reported in a sample of people with ME/CFS (29). Lymphoblasts from people with ME/CFS in one study, however, have not been observed to be depleted in mitochondrial DNA (30).

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Reference 26
Nguyen-Dien GT, Townsend B, Kulkarni PG, Kozul KL, Ooi SS, Eldershaw DN, et al. PPTC7 antagonizes mitophagy by promoting BNIP3 and NIX degradation via SCFFBXL4. EMBO Rep. 2024 Aug;25(8):3324–47.

Reference 27
Cao Y, Zheng J, Wan H, Sun Y, Fu S, Liu S, et al. A mitochondrial SCF-FBXL4 ubiquitin E3 ligase complex degrades BNIP3 and NIX to restrain mitophagy and prevent mitochondrial disease. EMBO J. 2023 Jul 3;42(13):e113033.

Reference 28
Bonnen PE, Yarham JW, Besse A, Wu P, Faqeih EA, Al-Asmari AM, et al. Mutations in FBXL4 cause mitochondrial encephalopathy and a disorder of mitochondrial DNA maintenance. Am J Hum Genet. 2013 Sep 5;93(3):471–81.

Reference 29
Tomas C, Brown A, Strassheim V, Elson JL, Newton J, Manning P. Cellular bioenergetics is impaired in patients with chronic fatigue syndrome. PLoS One. 2017;12(10):e0186802.

Reference 30
Missailidis D, Annesley SJ, Allan CY, Sanislav O, Lidbury BA, Lewis DP, et al. An Isolated Complex V Inefficiency and Dysregulated Mitochondrial Function in Immortalized Lymphocytes from ME/CFS Patients. Int J Mol Sci. 2020 Feb 6;21(3).

 

[Hutan]

Four of the eight loci (RABGAP1L, FBXL4, OLFM4,CA10) were associated at p < 0.05 with cases ascertained using post-exertional malaise and fatigue in the UK Biobank and the Netherlands biobank Lifelines.

So this was one locus that was replicated.

"OR is the Odds-Ratio of ME/CFS risk in cases versus controls." The OR for FBXL4 is less than 1, so I think that means that there is less chance of ME/CFS in people who have the variant i.e. the variant is protective.



I think this being a Tier 2 variant, it means that we don't know what the effect of the variant is? Does it up or down regulate FBXL4 and the mitophagy restriction?

FBXL4 regulates mitophagy, which is a process where surplus, aged or damaged mitochondria are degraded.

When FBXL4 is absent, the levels of BNIP3 and NIX raise and hyperactive mitophagy results. Loss-of-function mutations in FBXL4 are known to cause mitochondrial DNA depletion syndrome 13.

In DecodeME, the FBXL4 variant that was associated with ME/CFS was rs97984426. I couldn't find information on whether it would lead to icreased or decreased mitophagy.

In this study of mitochondrial DNA it was reported that "ME/CFS patients had an excess of individuals without a mildly deleterious population variant".
https://www.nature.com/articles/s41598-019-39060-1

Fewer deleterious mtDNA variants would suggest higher mitophagy is occurring. The question is how could this contribute to ME/CFS?

 

[Jonathan Edwards]

I wonder what this protein is regulated by - what changes it responds to. That might provide a clue.

 

[Jonathan Edwards]

It seems that the circadian rhythm gene CLOCK regulates this protein - might be relevant.

 

[V.R.T.]

It seems that the circadian rhythm gene CLOCK regulates this protein - might be relevant.

Have we seen this gene before in ME/CFS research?

Or am I just thinking of the Crawley pediatric LC study?

 

[arnoble]

I wonder what this protein is regulated by - what changes it responds to. That might provide a clue.

In the same section of CP's "Candidate" document, I am puzzling over this nearby miR-2113 locus:

"The interval also contains a non-protein long noncoding RNA locus (RP11-436D23.1) (25) which contains a miRNA locus (miR-2113) of unknown function. The allele that increases the risk of ME/CFS is associated with increasing RP11-436D23.1 expression in four tissues: amygdala, anterior cingulate cortex, cortex and hippocampus."

 

[sneyz]

Have we seen this gene before in ME/CFS research?

Or am I just thinking of the Crawley pediatric LC study?

Here’s something:

That's a really cool resource, thanks. For reference here is the paper the genes are from: Genetic Risk Factors for ME/CFS Identified using Combinatorial Analysis (Das et al, 2022, J Transl Med)

And here are the 14 genes they found, linked to their Protein Atlas cell type page (GC links to GeneCards page). I added where these proteins seem to be concentrated, just from a visual impression.
S100PBP - GC (glial, spermatocytes/spermatogonia)
ATP9A - GC (neuronal, glial)
KCNB1 - GC (neuronal)
CLOCK - GC (generally equal among cell types)
SLC15A4 - GC (dendritic)
TMEM232 - GC (excitatory/inhibitory neurons, glial, germ, ciliated)
GPC5 - GC (astrocytes)
PHACTR2 - GC (generally equal among cell types)
AKAP1 - GC (late spermatids)
USP6NL - GC (glial - mostly microglia)
CDON - GC (muller glia, excitatory/inhibitory neurons, mesothelial)
INSR - GC (generally equal among cell types)
SLC6A11 - GC (mainly astrocytes, but also other glial and neuronal cells)
SULF2 - GC (oligodendrocyte precursor, granulosa, endometrial stromal, maybe dendritic)

Edit: Also, just checked and there is no overlap between these genes and the 115 Zhang genes. The highest ranked of these is AKAP1 at position 1159 in the full Zhang list of 17759 genes.

 

[Jonathan Edwards]

Have we seen this gene before in ME/CFS research?

Yes, people have commented on it before, I think in relation to unrefreshing sleep.

 

[chillier]

Have we seen this gene before in ME/CFS research?

Or am I just thinking of the Crawley pediatric LC study?

CLOCK was one of the genes found in precisionLife's ME/CFS genetics study in the UK biobank, and replicated later in a long covid cohort

 

[Hutan]

I think a paper from Tate's group, Sweetman? found something with the CLOCK gene/control of circadian rhythm.