ZSCAN25 methylation predicts seizures and severe alcohol withdrawal syndrome

Figures & data

Table 1. Clinical characteristics of the subjects whose data were used to identify candidate markers.

	Cases		Abstinent Controls
Male	Female	Male	Male	Female
N	35	10	30	17
Age	43 ± 12	42 ± 10	48 ± 15	41 ± 11
Ethnicity
European American	31	9	28	17
African American	4	1	2
Drinks per day over past month	14.4 ± 9.5	15.6 ± 13.4	-	-
Days between sampling	21.8 ± 4.7	22.3 ± 4.7	-	-

Table 2. Leading candidates from initial screening efforts to identify rapidly reverting loci.

Illumina ID	Gene	T1-T2 Difference	P-Value	Bonferroni	2014 T1-T2 Difference^2
cg27546431	MYO1C	0.074213	1.14E–05	NS	0.010645
cg09807356	GALNS	0.066176	2.09E–10	0.000178	0.028749
cg12389043	CCDC92	0.052147	3.31E–07	NS	0.013004
cg07692435		0.051218	4.34E–06	NS
cg07375256	ZSCAN25 [1]	0.051036	3.28E–07^a	NS	0.056552
cg18424841		0.049764	0.000133	NS	0.034717
cg21281638	C7orf26	0.046891	1.26E–05	NS

^aZSCAN25 is previously referred to as ZNF498.

^bThe values for 2014 T1-T2 difference are taken from supplemental table S2 in Philibert et al., 2014.

Figure 1. The correlation between the Illumina probe assessment and the MSdPCR assessments at the Alc12 locus (n = 90) using the Dcg07375256 assay. Overall, the r = 0.92 with the dynamic range of the ddPCR being considerably greater than that of the methylation array.

Figure 2. Methylation at cg07375256 in HAC subjects at study intake (T1) and exit (T2), and in abstinent controls (n = 46, all groups).

Table 3. Clinical characteristics of alcohol withdrawal subjects and new controls.

	Cases		Controls
	Female	Male	Male	Female
N	32	93	67	70
Age in years (SD)	47.4 ± 15.4	44.4 ± 13.3	43.9 ± 14.3	39.0 ± 17.0
Hispanic Race (may be > one)	1	1	5	4
Native American	1	5	0	0
Asian	0	0	5	3
African American	1	3	2	2
Hawaiian	0	0	0	0
European American	31	87	61	64
Other	0	2	0	1
Prefer not to answer	0	1	0	1
Lifetime ≥100 cigarettes	13	60	-	-
Smoked ≥1 PPD in last month	4	22	1	0
Drinks per day in past month
0 to 4	10	12	65	68
5 to 9	8	12	2	1
10 to 14	5	25	0	1
15 to 19	2	18	0	0
20 or more	6	26	0	0
Not reported	1	0	0	0
Median days since last drink	2	2	-	-
MAST Score (SD)	-	-	2.5 ± 1.0	3.3 ± 1.9
Key Outcomes
No BZDs given	6	26	-	-
BZD given	26	77	-	-
Phenobarbital given	2	17	-	-
Seizures	0	7	-	-
Hallucinations	4	11	-	-
ICU Admission	0	8	-	-
Delirium Tremens	0	8	-	-

Figure 3. The correlation of study variables to each other. * = p < 0.05, ** p < 0.01, *** p < 0.001. Age is expressed in years. Sex is binary variable whether or not the subject is male. Drinks scale is an ordinal variable with higher score indicating greater intake. PAWSS is prediction of alcohol withdrawal syndrome scale. BAC > 200 indicates if a breathalyser value greater than 200 mg/dl was noted at admission. CDT is carbohydrate transferrin levels. Cg05575921 is DNA methylation at that smoking related locus. ATS is alcohol T score. ZSCAN25 is DNA methylation per the Dcg07375256 described herein. Diazepam equivalents is the sum total equivalent of all benzodiazepines administered during hospitalization for AWS. Phenobarbital is a binomial variable indicating whether or not phenobarbital was given during hospitalization for AWS. Seizures, hallucinations, admission to the ICU, and delirium tremens (DTs) are binomial variable for the presence of those symptoms.

Figure 4. The relationship of predictor (PAWSS, ATS, CDT and ZSCAN (Dcg07375256)) values to treatment group. ZSCAN25 methylation and CDT values are given in percent. ATS and PAWSS have no unit values. Because control subjects were not administered the PAWSS, their values are not considered in that section of the analyses. Kruskal-Wallis p-values are given for each indicated contrasts.

Figure 5. The relationship between predictor (PAWSS, ATS, CDT and ZSCAN (Dcg07375256)) levels and presence (1) or absence (0) of seizures. N = 7 for the seizure group, 116 for the no seizure group. ZSCAN25 methylation and CDT values are given in percent. ATS and PAWSS have no unit values. Kruskal-Wallis p-values are given for each indicated contrasts.

Academic ED SBIRT Research Collaborative. The impact of screening, brief intervention, and referral for treatment on emergency department patients’ alcohol use. Ann Emerg Med. 2007; 50:699–710. e696. doi: 10.1016/j.annemergmed.2007.06.486

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