1. Medicine

Metabolomics identifies and validates serum androstenedione as novel biomarker for diagnosing primary angle closure glaucoma and predicting the visual field progression

  1. Shengjie Li  Is a corresponding author
  2. Jun Ren
  3. Zhendong Jiang
  4. Yichao Qiu
  5. Mingxi Shao
  6. Yingzhu Li
  7. Jianing Wu
  8. Yunxiao Song
  9. Xinghuai Sun
  10. Shunxiang Gao  Is a corresponding author
  11. Wenjun Cao  Is a corresponding author
  1. Department of Clinical Laboratory, Eye & ENT Hospital, Shanghai Medical College, Fudan University, China
  2. Department of Ophthalmology & Visual Science, Eye & ENT Hospital, Shanghai Medical College, Fudan University, China
  3. State Key Laboratory of Medical Neurobiology, Institutes of Brain Science, Fudan University, China
  4. Key Laboratory of Myopia, Chinese Academy of Medical Sciences, China
  5. NHC Key Laboratory of Myopia, Fudan University, China
  6. Department of Clinical Laboratory, Shanghai Xuhui Central Hospital, Fudan University, China
  7. Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, China
  8. National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, China
https://doi.org/10.7554/eLife.91407.3
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Cite this article
  1. Shengjie Li
  2. Jun Ren
  3. Zhendong Jiang
  4. Yichao Qiu
  5. Mingxi Shao
  6. Yingzhu Li
  7. Jianing Wu
  8. Yunxiao Song
  9. Xinghuai Sun
  10. Shunxiang Gao
  11. Wenjun Cao
(2024)
Metabolomics identifies and validates serum androstenedione as novel biomarker for diagnosing primary angle closure glaucoma and predicting the visual field progression
eLife 12:RP91407.
https://doi.org/10.7554/eLife.91407.3
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10 figures, 2 tables and 11 additional files

Figures

Figure 1 with 2 supplements
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Study design and workflow.

A five-phase study (discovery phase [discovery set 1, the discovery set 2], validation phase 1, validation phase 2, supplemental phase, and cohort phase) design. (A) The workflow of the discovery …

Figure 1—figure supplement 1
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Locations of the eight districts in China.

(A) All the primary angle closure glaucoma (PACG) patients; (B) discovery set 1; (C) discovery set 2; (D) validation phase 1; (E) validation phase 2.

Figure 1—figure supplement 2
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The workflow of mass spectrometry.
Figure 2
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Metabolic profiles discriminate participants with primary angle closure glaucoma (PACG) from normal controls (NC).

(A) Orthogonal projection to latent structure-discriminant analysis (OPLS-DA) score plot of the comparison between the PACG and NC groups in the discovery phase (discovery set 1 and discovery set …

Figure 3 with 2 supplements
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Identification of a unique primary angle closure glaucoma (PACG)-associated blood metabolite fingerprint and its behavior in the discovery phase.

(A) Heatmap of the area under the receiver operating characteristic curve assessing the discriminating accuracy of each of the 32 metabolites in differentiating PACG from normal control in the …

Figure 3—figure supplement 1
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The level of the 31-metabolite cluster between primary angle closure glaucoma (PACG) and control patients in discovery set 1 (unit for y-axis is peak areas).
Figure 3—figure supplement 2
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The level of the 31-metabolite cluster between primary angle closure glaucoma (PACG) and control patients in discovery set 2 (unit for y-axis is peak areas).
Figure 4 with 2 supplements
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Biomarker discovery discriminates primary angle closure glaucoma (PACG) from normal in the discovery phase.

(A) A heatmap of correlation analysis between ocular clinical characteristics and 32 potential biomarkers in the discovery phase in PACG subjects. (B) The serum level of androstenedione between PACG …

Figure 4—figure supplement 1
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Heatmap of correlation analysis between ocular clinical characteristics and 32 potential biomarkers in discovery set 1 of primary angle closure glaucoma (PACG) subjects.
Figure 4—figure supplement 2
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Heatmap of correlation analysis between ocular clinical characteristics and 32 potential biomarkers in discovery set 2 of primary angle closure glaucoma (PACG) subjects.
Figure 5 with 2 supplements
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Biomarker validation in two independent validation phases to discriminate primary angle closure glaucoma (PACG) from normal.

(A) The serum level of androstenedione between PACG and normal group in validation phase 1 (unit for y-axis is peak areas). (B) The serum level of androstenedione between PACG and normal group in …

Figure 5—figure supplement 1
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Orthogonal projection to latent structure-discriminant analysis (OPLS-DA) score plot of the comparison between the primary angle closure glaucoma (PACG) and normal control (NC) groups in validation phase 1.
Figure 5—figure supplement 2
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Biomarker validation in male and female subgroups.

(A) Androstenedione to discriminate primary angle closure glaucoma (PACG) from normal in the discovery set in male subjects. (B) Androstenedione to discriminate PACG from normal in the discovery set …

Figure 6 with 1 supplement
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Androstenedione is associated with the severity of primary angle closure glaucoma (PACG).

(A) Comparison of mean serum levels of androstenedione between mild, moderate, and severe PACG in the discovery set 1 (unit for y-axis is peak areas). (B) Comparison mean serum levels of …

Figure 6—figure supplement 1
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Receiver operating characteristic curves to discriminates mild, moderate, and severe primary angle closure glaucoma (PACG).

(A) Androstenedione to discriminates mild, moderate, and severe PACG in discovery set 1. (B) Androstenedione to discriminates mild, moderate, and severe PACG in discovery set 2. (C) Androstenedione …

Figure 7
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Specificity of circulating androstenedione in patients with primary angle closure glaucoma (PACG) in supplemental phase.

(A) Sampling scheme and workflow to investigate the temporal changes in androstenedione levels. (B) Differential level of serum androstenedione between patients with PACG before and 3 months after …

Figure 8
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Calibration ability of androstenedione.

(A) Calibration curves in the discovery set 1. (B) Calibration curves in the discovery set 2. (C) Calibration curves in the validation phase 1. (D) Calibration curves in the validation phase 2. The …

Figure 9
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Kaplan-Meier curves stratified by the men value in terms of androstenedione.

(A) Male+female; (B) female; (C) male. We categorized study participants into two groups based on their mean levels of androstenedione.

Figure 10 with 2 supplements
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Potential mechanisms, androstenedione accumulation in patients with primary angle closure glaucoma (PACG).

(A) Steroid hormone biosynthesis related to androstenedione. (B) Aromatase function deficits may be the potential mechanism leading to androstenedione accumulation in patients with PACG. (C) Binding …

Figure 10—figure supplement 1
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Pathway enrichment plot in discovery set 2.
Figure 10—figure supplement 2
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Molecular formula of androstenedione.

Tables

Table 1
The clinical and demographic characteristics of all subjects in the discovery and validation phases.
Normal (n=268)PACG (n=348)t/χ2p
Discovery phase
Discovery set 1
Number (n)6080
Age (years)58.75±8.6361.00±8.671.520.13
Sex (male, %)24 (40.0)31 (38.8)0.020.88
BMI (kg/m2)24.04±5.2723.36±2.440.990.33
Hypercholesterolemia (yes, %)6 (10)9 (11.3)0.060.81
Hypertension (yes, %)16 (26.7)18 (22.8)0.320.60
Diabetes (yes, %)1 (1.7)8 (10.1)3.960.10
Smoking (yes, %)5 (8.3)11 (13.9)0.990.42
Drinking (yes, %)7 (11.7)8 (10.1)0.10.76
Duration (months)10.45±12.43
IOP (mmHg)12.22±4.5027.78±11.2010.17<0.001
VCDR0.25±0.180.64±0.2310.87<0.001
AL (mm)22.48±1.14
ACD (mm)1.86±0.55
CCT (µm)534.04±41.26
MS (dB)12.46±8.75
MD (dB)14.67±8.94
Discovery set 2
Number (n)80100
Age (years)62.54±6.7463.14±9.040.490.62
Sex (male, %)35 (43.8)33 (33.0)2.190.14
BMI (kg/m2)23.68±2.8123.32±3.151.360.18
Hypercholesterolemia (yes, %)12 (15)13 (13)0.150.70
Hypertension (yes, %)26 (32.5)33 (33.0)0.010.94
Diabetes (yes, %)7 (8.8)9 (9.0)0.0030.95
Smoking (yes, %)11 (13.8)15 (15.0)0.060.81
Drinking (yes, %)19 (23.8)22 (22.0)0.080.78
Duration (months)8.46±9.69
IOP (mmHg)11.45±5.2125.80±12.558.329<0.001
VCDR0.27±0.140.61±0.2210.48<0.001
AL (mm)22.43±0.75
ACD (mm)2.05±0.77
CCT (µm)547.43±43.04
MS (dB)12.34±8.55
MD (dB)14.50±8.95
Validation phase 1
Number (n)5070
Age (years)57.47±8.1760.34±10.111.660.10
Sex (male, %)18 (36.0)24 (34.3)0.040.85
BMI (kg/m2)23.08±1.9924.45±3.721.310.19
Hypercholesterolemia (yes, %)4 (8)8 (11.4)0.380.54
Hypertension (yes, %)17 (34.0)21 (30.0)0.220.64
Diabetes (yes, %)5 (10.0)7 (10.0)0.01.0
Smoking (yes, %)6 (12.0)10 (14.3)0.130.72
Drinking (yes, %)10 (20.0)15 (21.4)0.040.85
Duration (months)11.09±13.82
IOP (mmHg)12.90±4.1128.46±9.8011.55<0.001
VCDR0.24±0.170.60±0.2110.87<0.001
AL (mm)22.44±0.83
ACD (mm)1.86±0.37
CCT (µm)539.09±82.70
MS (dB)13.78±8.58
MD (dB)13.96±9.29
Validation phase 2
Number (n)7898
Age (years)56.55±11.5360.26±15.411.770.08
Sex (male, %)29 (37.2)48 (49.0)2.460.12
BMI (kg/m2)24.54±5.4225.90±7.511.260.21
Hypercholesterolemia (yes, %)11 (14.1)13 (13.3)0.030.87
Hypertension (yes, %)19 (24.4)31 (31.6)0.260.61
Diabetes (yes, %)6 (7.7)17 (17.3)3.60.06
Smoking (yes, %)9 (11.5)22 (22.4)3.60.06
Drinking (yes, %)12 (15.4)19 (19.4)0.480.49
Duration (months)11.40±13.8
IOP (mmHg)13.40±5.4329.20±11.2010.07<0.001
VCDR0.30±0.190.68±0.2310.41<0.001
AL (mm)23.20±1.72
ACD (mm)2.15±0.67
CCT (µm)546.71±50.35
MS (dB)11.07±8.54
MD (dB)16.49±8.64

  1. BMI = body mass index; IOP = intraocular pressure, VCDR = vertical cup-to-disc ratio, AL = axial length, CCT = central corneal thickness, ACD = anterior chamber depth, MD: visual field mean deviation, MS: visual field mean sensitivity, PACG = primary angle closure glaucoma.

Table 2
Cox proportional hazards regression analysis to assess the value of androstenedione associated with progression of PACG.
UnivariateMultivariate*
pHR (95% CI)pHR (95% CI)
Age0.0080.97 (0.95–0.99)0.220.98 (0.95–1.01)
Sex0.870.95 (0.53–1.72)0.731.15 (0.53–2.52)
IOP0.591.007 (0.98–1.04)0.750.99 (0.96–1.03)
VCDR0.170.40 (0.11–1.46)0.160.37 (0.092–1.46)
CCT0.771.001 (0.10–1.007)0.901.000 (0.99–1.007)
ACD0.031.63 (1.05–2.52)0.911.041 (0.51–2.13)
AL0.0051.24 (1.07–1.44)0.351.12 (0.89–1.41)
MD0.131.04 (0.99–1.08)0.0751.047 (0.10–1.10)
Androstenedione<0.0013.73 (1.84–7.57)0.0172.71 (1.20–6.10)
  1. *

    Adjusted for BMI, diabetes (yes = 1, no = 0), hypertension (yes = 1, no = 0), hypercholesterolemia (yes = 1, no = 0), smoking (yes = 1, no = 0), and drinking (yes = 1, no = 0). Bold values indicate positive results.

  2. IOP =intraocular pressure, VCDR =vertical cup-to-disc ratio, AL =axial length, CCT =central corneal thickness, ACD =anterior chamber depth, MD =mean deviation, PACG =primary angle closure glaucoma.

Additional files

Supplementary file 1

List of statistical approach and packages.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp1-v1.docx
Supplementary file 2

The clinical and demographic characteristics of primary angle closure glaucoma (PACG) and cataract subjects in the supplemental phase.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp2-v1.docx
Supplementary file 3

The clinical and demographic characteristics of same primary angle closure glaucoma (PACG) patients between pre-treatment and post-treatment.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp3-v1.docx
Supplementary file 4

The differential metabolites associated with primary angle closure glaucoma (PACG) and their fold changes in discovery set 1.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp4-v1.docx
Supplementary file 5

The differential metabolites associated with primary angle closure glaucoma (PACG) and their fold changes in discovery set 2.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp5-v1.docx
Supplementary file 6

The detecting parameters of 32 differential metabolites.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp6-v1.docx
Supplementary file 7

The relationship between androstenedione and risk of primary angle closure glaucoma (PACG).

https://cdn.elifesciences.org/articles/91407/elife-91407-supp7-v1.docx
Supplementary file 8

The results of receiver operating characteristic (ROC) curve.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp8-v1.docx
Supplementary file 9

Comparison of characteristics of no progression and progression group in primary angle closure glaucoma (PACG) patients.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp9-v1.docx
Supplementary file 10

Comparison of areas under the receiver operating characteristic curve (AUCs) value among docosahexaenoic acid (DHA), free fatty acid (FFA) (22:6), free fatty acid (FFA) (18:4), and androstenedione.

https://cdn.elifesciences.org/articles/91407/elife-91407-supp10-v1.docx
MDAR checklist
https://cdn.elifesciences.org/articles/91407/elife-91407-mdarchecklist1-v1.docx

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