CN-122029433-A - Methods, systems and kits for detecting retinopathy-related proteomes in premature infants

CN122029433ACN 122029433 ACN122029433 ACN 122029433ACN-122029433-A

Abstract

A method for detecting the expression level of retinopathy-related proteomes in premature infants is provided. The method comprises obtaining a tear sample from a premature infant and obtaining a protein profile from the tear sample, detecting a set of retinopathy-related proteomes comprising one or more up-regulating proteins and down-regulating proteins in the protein profile, calculating a Log 2 FC for each up-regulating protein relative to its baseline expression level and similarly calculating a Log-double change for each down-regulating protein, and generating a list comprising up-regulating proteins having a Log-double change greater than or equal to a positive threshold and down-regulating proteins having a Log-double change less than or equal to a negative threshold.

Inventors

LIN WEIQING
LIN QUAN
SHI YINGHAN
Liao Kaiying
Zheng Baiwen

Assignees

眼视觉研究中心有限公司
香港理工大学

Dates

Publication Date: 20260512
Application Date: 20250911
Priority Date: 20240911

Claims (14)

1. A method of diagnosing retinopathy of prematurity in a premature infant by detecting and calculating the expression level of a retinopathy-related proteome of premature infant, comprising: Obtaining a tear sample from the premature infant; Extracting a protein profile from the tear sample; detecting the retinopathy-related proteome in the protein profile, which includes one or more up-regulating proteins and down-regulating proteins; calculating a logarithmic double change value of each of the up-regulated proteins with respect to a reference expression amount thereof, and similarly calculating a logarithmic double change value of each of the down-regulated proteins, and Generating a protein list as a molecular signature of a retinopathy-related tear biomarker, wherein the protein list comprises: The logarithmic doubling change value is greater than or equal to the positive threshold, and Down-regulating protein with logarithmic doubling change value smaller than or equal to negative threshold value; Wherein the up-regulating protein comprises one or more of beta 2 microglobulin, fatty acid binding protein 5, protein S100-A4, secretoglobin family 1D member 1, interleukin-1 receptor antagonist, glutathione reductase, GTP binding nucleoprotein Ran, eukaryotic initiation factor 4A-I, polymeric immunoglobulin receptor, NAD (P) H dehydrogenase 1, cysteine-rich protein 1, membrane-associated phospholipase A2, DNA dC→dU editing enzyme APOBEC-3A, chondroitin 1, C-X-C motif chemokine 17, and the down-regulating protein comprises one or more of antithrombin III, fructose-1, 6-bisphosphatase 1, apolipoprotein D, apolipoprotein A-I, apolipoprotein A-IV, adenylate kinase isozyme 1, beta-hexosaminidase alpha subunit, hemochrome binding protein, nail protein, serum transferrin, AMBP protein, inter-heavy chain H2, thyroxine 4-H-alpha-glycoprotein, ganglioside 2, alpha-fetoprotein, and alpha-2H-fetoprotein.
2. The method of claim 1, wherein the positive threshold is 0.58.
3. The method of claim 1, wherein the negative threshold is-0.58.
4. The method of claim 1, wherein the baseline performance is obtained by detecting relative protein in a tear sample of a healthy premature infant.
5. The method of claim 1, wherein the protein profile is acquired by a data independent acquisition mass spectrometer and quantified by a machine learning based analysis step.
6. A system for diagnosing retinopathy of prematurity in a premature infant by detecting and calculating the expression level of a retinopathy-related proteome of premature infant, comprising: A sample collection module configured to collect a tear sample of a premature infant; A protein analysis module configured to extract a protein profile from the tear sample; A protein detection module configured to detect the expression level of a set of retinopathy-related proteomes of premature infants in the protein profile, wherein the proteomes include one or more up-regulating proteins and one or more down-regulating proteins, and A computing module configured to: Calculating a logarithmic double change value of each up-regulated protein relative to a reference expression quantity; Calculating the logarithmic double change value of each of the down-regulated proteins relative to the reference expression amount, and Generating a protein list as a molecular feature of a retinopathy-related tear biomarker to facilitate subsequent clinical decision making or risk stratification, wherein the protein list comprises: An up-regulated protein having a logarithmic doubling change value greater than or equal to a positive threshold, and The log double change value is less than or equal to a negative threshold.
7. The system of claim 6, wherein the up-regulating protein comprises one or more of beta 2 microglobulin, fatty acid binding protein 5, protein S100-A4, secretoglobin family 1D member 1, interleukin-1 receptor antagonist, glutathione reductase, GTP-binding nucleoprotein Ran, eukaryotic initiation factor 4A-I, polymeric immunoglobulin receptor, NAD (P) H dehydrogenase 1, cysteine-rich protein 1, membrane-associated phospholipase A2, DNA dC→dU editing enzyme APOBEC-3A, chondrogenic protein 1, C-X-C motif chemokine 17.
8. The system of claim 6, wherein the down-regulating protein comprises one or more of antithrombin III, fructose-1, 6-bisphosphatase 1, apolipoprotein D, apolipoprotein A-I, apolipoprotein A-IV, adenylate kinase isozyme 1, beta-hexosaminidase alpha subunit, hemoglobin binding protein, alpha fetoprotein, serum transferrin, AMBP protein, an inter-trypsin inhibitor heavy chain H2, transthyretin, retinol binding protein 4, apolipoprotein H, alpha-2-HS-glycoprotein, ganglioside GM2 activator protein.
9. The system of claim 6, wherein the baseline performance is obtained by detecting relative protein in a tear sample of a healthy premature infant.
10. The system of claim 6, wherein the positive threshold is 0.58 and the negative threshold is-0.58.
11. The system of claim 6, wherein the protein analysis module comprises a data independent acquisition mass spectrometer.
12. A kit for detecting a retinopathy-related proteome of premature infants, comprising: a protein assay for detecting the presence or amount of retinopathy-related proteomes of premature infants in a biological sample of premature infants; Wherein the retinopathy-related proteome of premature infants comprises beta 2 microglobulin, fatty acid binding protein 5, protein S100-A4, secretoglobin family 1D member 1, interleukin-1 receptor antagonist, glutathione reductase, GTP binding nucleoprotein Ran, eukaryotic initiation factor 4A-I, polymeric immunoglobulin receptor, NAD (P) H dehydrogenase 1, cysteine-rich protein 1, membrane-associated phospholipase A2, DNA dC→dU editing enzyme APOBEC-3A, chondrotin 1, C-X-C motif chemokine 17, antithrombin III, fructose-1, 6-bisphosphatase 1, apolipoprotein D, apolipoprotein A-I, apolipoprotein A-IV, adenylate kinase isozyme 1, beta-hexosaminidase alpha subunit, hemoglobin binding protein, alpha fetoprotein, serum transferrin, AMBP protein, inter-inhibitor heavy chain H2, transferrin, retinol binding protein 4, alpha-2H, alpha-2-ganglioside, trypsin-2, and ganglioside-activated protein.
13. The kit of claim 12, wherein the protein detection method comprises mass spectrometry detection, antibody immunoassay, protein chip, or aptamer detection system.
14. The kit of claim 12, wherein the biological sample comprises a tear, blood, serum, or plasma sample.

Description

Methods, systems and kits for detecting retinopathy-related proteomes in premature infants Cross Reference to Related Applications The present application claims priority from U.S. provisional patent application No. 63/693,209 filed on day 11, 9, 2024 and U.S. provisional patent application No. 63/767,518 filed on day 5, 3, 2025, the disclosures of which are incorporated herein by reference in their entirety. Technical Field The present invention relates to the field of medical science and biological information. More particularly, the present invention relates to methods, systems and kits for detecting retinopathy-related proteomes in premature infants. Background Retinopathy of prematurity is a severe ophthalmic disease characterized by abnormal proliferation of retinal blood vessels in premature infants. The pathological neovasculature usually occurs at the junction of the vascular and avascular regions of the immature retina and may further form fibrous scar tissue. This shrinkage of the fibrotic tissue may eventually lead to retinal detachment, resulting in permanent vision loss. Thus, retinopathy of prematurity is recognized as a major cause of global preventable childhood blindness. With the progress of neonatal care, the survival rate of premature infants is higher and higher, and the occurrence rate of retinopathy of prematurity in the world is also increased remarkably. Despite the clinical relevance of retinopathy of prematurity, its underlying etiology and pathophysiological mechanisms are not fully understood. The retinopathy of prematurity progresses rapidly and seriously, and clinically, the retinopathy of prematurity can be divided into five stages, namely, a slight dividing line between blood vessels and non-blood vessel retinas occurs in the first stage, the formation of ridge-like protrusion in the second stage, pathological angiogenesis proliferation occurs in the third stage, partial retinal detachment in the fourth stage and complete retinal detachment in the fifth stage. In addition, retinopathy of prematurity can be divided into three concentric retina areas with optic disc as center according to focus position type, wherein the area I is the innermost circle and comprises macula part and optic disc, the area II is the middle peripheral retina, and the area III is the crescent peripheral retina. Timely identification of disease progression is critical because early intervention with anti-Vascular Endothelial Growth Factor (VEGF) therapy or laser photocoagulation can effectively prevent the progression of disease and maintain vision. How to find the progress of the disease in time is important, because the disease can be controlled and the vision can be protected by early intervention of anti-vascular endothelial growth factor treatment or laser photocoagulation. Currently, indirect ophthalmoscopy is clinically used as the gold standard for screening retinopathy of prematurity. However, this method requires specialized equipment and is highly dependent on subjective interpretation by experienced ophthalmologists, and therefore, the interpretation of subtle retinal changes by different clinicians tends to vary, and the diagnosis of disease stages may also vary, often resulting in inconsistent diagnosis. Furthermore, the examination procedure often requires repeated dilation of the pupil with a ciliary paralytic or mydriatic agent, which can create a physiological burden for the fragile premature infant and cause side effects. Although imaging screening methods for retinopathy of prematurity exist, the prognosis is limited and is usually only detected after the onset of the disease and cannot be effectively predicted or assessed at early risk. Thus, many premature infants fail to get treatment in time, resulting in irreversible retinal damage. Thus, there is a great clinical need for a reliable and non-invasive diagnostic test technique for the detection or prognosis of retinopathy of prematurity via measurable biomarkers. Traditional diagnostic methods, such as direct sampling of retinal tissue, are highly invasive and highly risky, and can result in permanent retinal damage. The current prior art does not accurately predict the likelihood of retinopathy of prematurity in high risk premature infants. Thus, the present invention provides a non-invasive biomarker-based protocol to address this clinical need and to detect or predict retinopathy of prematurity in an early stage, so as to facilitate a clinically advanced treatment and reduce the risk of blindness. Disclosure of Invention The invention aims to provide a method and a system for solving the technical problems. According to a first aspect of the present invention there is provided a method of detecting the retinopathy of prematurity associated proteome performance of a premature infant, the method comprising taking a tear sample from the premature infant, extracting a protein profile from the tear sample, and detecting a retino