Determination of Blood NOTCH3 Extracellular Domain and Jagged-1 Levels in Healthy Subjects

Abstract

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common genetic disorder among those responsible for hereditary strokes, and it is caused by a mutation in the NOTCH3 gene on chromosome 19. Blood biomarkers related to the Notch signaling pathway have not been investigated extensively in CADASIL. In this study, we measured the serum and plasma levels of NOTCH3 extracellular domain (N3ECD) and its ligand, Jagged-1, in 279 healthy subjects. The levels of N3ECD and Jagged-1 showed significant correlations in both serum (p < 0.0001, r = 0.2681) and plasma (p < 0.0001, r = 0.4065). The N3ECD levels were significantly higher in the serum than in plasma and tend to increase with age. In contrast, there was no significant difference between the serum and plasma levels of Jagged-1 levels. To summarize, we were able to measure N3ECD and Jagged-1 protein levels in healthy human serum and plasma. Taken together, our findings provide the basis for further studies investigating the clinical use of blood N3ECD and Jagged-1 levels for CADASIL and other Notch signaling-related diseases.

Keywords: CADASIL; Jagged-1; NOTCH3 extracellular domain; blood marker; plasma; serum.

Figure 1

NOTCH3 extracellular domain (N3ECD) levels in healthy adults. N3ECD levels were measured in serum and plasma from healthy adults by ELISA (n = 279) (A) and a correlation between serum and plasma N3ECD levels was analyzed (B). Jagged-1 levels were measured in serum and plasma (C) and their correlation was assessed (D). Each symbol represents a value from a single subject; midline represents the mean; and error bars represent the standard error. The resulting Spearman’s correlation coefficient (r) and corresponding p value are reported.

Figure 2

Correlations between N3ECD and Jag-1 levels. Positive correlations of serum (A) and plasma levels (B) between N3ECD and Jag1 (n = 279). Results are representative of three independent ELISA experiments. The resulting Spearman’s correlation coefficient (r) and corresponding p-values are reported.

Figure 3

Comparison of mean N3ECD and Jagged-1 levels by gender and age. Serum and plasma N3ECD levels between male and female (A) and correlations between N3ECD levels and age (B). Jagged-1 serum and plasma levels in male and female (C) and correlations with age (D). Each symbol represents values from a single subject; midline represents the mean; and error bars represent the standard error. The resulting Spearman’s correlation coefficient (r) and corresponding p-value are reported. ns—not significant.

Figure 4

Correlations between serum N3ECD levels and laboratory values. Correlations of N3ECD with creatinine (A), and hemoglobin (B). Correlations of serum Jag-1 with systolic blood pressure (C), HbA1c (D), and triglyceride (E). Each symbol represents values from a single subject. The resulting Spearman’s correlation coefficient (r) and corresponding p value are reported.