Exploring urinary modified nucleosides as biomarkers for diabetic retinopathy: Development and validation of a ultra performance liquid chromatography-tandem mass spectrometry method

  • J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Jan 1:1232:123968. doi: 10.1016/j.jchromb.2023.123968.
Chen Yao  1 ,  Daizhu Lv  2 ,  Xueqing Zhou  3 ,  Pengcheng Fu  4 ,  Wen Sun  5 ,  Jinlian Chen  6 ,  Huan Lin  7
Affiliations
  • 1. State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou 570228, PR China; School of Life Sciences, Hainan University, Haikou 570228, PR China. Electronic address: [email protected].
  • 2. Analysis and Testing Center, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, PR China. Electronic address: [email protected].
  • 3. Analysis and Testing Center, Hainan University, Haikou 570228, PR China. Electronic address: [email protected].
  • 4. State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou 570228, PR China. Electronic address: [email protected].
  • 5. Hainan Traditional Chinese Medicine Hospital, Hospital of Chinese Medicine Affiliated by Hainan Medical College, Haikou 570203, PR China. Electronic address: [email protected].
  • 6. Hainan Eye Hospital and Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Haikou 570311, PR China. Electronic address: [email protected].
  • 7. State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou 570228, PR China. Electronic address: [email protected].
PMID: 38150972 DOI: 10.1016/j.jchromb.2023.123968
Abstract

The dynamic modification of RNA plays a crucial role in biological regulation and is strongly linked to human disease development and progression. Notably, modified nucleosides in urine have shown promising potential as early diagnostic biomarkers for various conditions. In this study, we developed and validated a rapid, sensitive, and accurate UPLC-MS/MS method for quantifying eight types of modified nucleosides (N1-methyladenosine (m1A), N6-methyladenosine (m6A), 5-methyluridine (m5U), 5-taurinomethyl-2-thiouridine (τm5s2U), 5-methylcytidine (m5C), 2'-O-methylcytidine (Cm), N1-methylguanosine (m1G), and N7-methylguanosine (m7G) in human urine. Using the method, we measured the urinary concentrations of m1A, m6A, m5U, τm5s2U, m5C, Cm, m1G, and m7G in a total of 21 control individuals and 23 patients diagnosed with diabetic retinopathy (DR). Cm levels showed promise as a diagnostic marker for diabetic retinopathy (DR), with a significant value (P < 0.01) and an AUC of 0.735. Other modified nucleosides also exhibited significant differences within specific subpopulations. As non-proliferative diabetic retinopathy (NPDR) signifies the latent early stage of diabetic retinopathy, we developed a multivariate linear model that integrates patients' sex, age, height, and urinary concentration of modified nucleosides which aims to predict and differentiate between healthy individuals, NPDR patients, and proliferative diabetic retinopathy (PDR) patients. Encouragingly, the model achieved satisfactory accuracy rates: healthy (81%), NPDR (75%), and PDR (80%). Our findings provide valuable insights into the development of an early, cost-effective, and noninvasive diagnostic approach for diabetic retinopathy.

Keywords
Biomarkers; Diabetic retinopathy; Modified nucleosides; UPLC-MS/MS.
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