Urine accommodates more changes than other fluids, and it is a good source in the search for early sensitive biomarkers. The present study collected urine samples from 2-, 4-, 6-, 8-and 10-month-old αsynuclein transgenic mice. Based on data-independent acquisition (DIA) technology, liquid chromatographytandem mass spectrometry (LC-MS/MS) was used for quantitative analysis. Seventeen human homologous differential proteins were screened and compared with those in the urine of 2-month-old mice, and 9 proteins were related to Parkinson's disease (PD). Formin-2, Splicing factor 3A subunit 1, and Isopentenyl-diphosphate Deltaisomerase 1 changed continuously in months 6, 8 and 10. These experiments and analyses demonstrated that the urine proteome reflected the development of α-synuclein transgenic mice and provided clues for the early clinical diagnosis of PD. Parkinson's disease (PD) is a neurodegenerative disease characterized by progressive motor and non-motor disorders [1]. Its main pathological features are the progressive loss of dopaminergic neurons in the dense substantia nigra and the presence of α-synuclein as the main component of eosinophilic inclusion bodies, i.e., Lewy bodies, in the remaining dopaminergic neurons. PD is the second most common neurodegenerative disease, and it affects approximately 1% of people over 65 years of age [2]. With the aging of the population and the prolongation of life expectancy, the incidence rate of PD is expected to double in the next 20 years [3]. PD seriously affects the quality of life of patients, and it results in a huge social and economic burden [4, 5]. The early diagnosis of PD is of great significance to improve the quality of life of patients and delay the development of the disease. There is no effective way to cure PD. Drug therapy and surgical treatment only delay the progress of the disease. Drug therapy is the primary method, but the long-term use of drugs often weakens the curative effects and produces some side effects. The clinical diagnosis of PD is currently based on motor characteristics. When the motor characteristics of PD manifest, approximately 60-70% of the neurons are lost [6]. Because the early symptoms of PD are mild and generally considered the result of aging, the early diagnosis of PD is very difficult, and there is a certain rate of misdiagnosis. At the time of diagnosis, most patients exhibit obvious behavior disorders and accumulated pathological changes. Therefore, to provide better clinical treatment for PD patients and help researchers find new treatment methods, it is urgent to identify sensitive early biomarkers. Biomarkers are indicators that objectively reflect normal physiological and pathological processes [7]. The most studied biomarkers are present in blood and cerebrospinal fluid. Because there is no barrier between the cerebrospinal fluid and the brain, cerebrospinal fluid is considered an ideal choice for the discovery of biomarkers of neurodegenerative diseases. Cerebrospinal fluidis more stable than blood due to the blood-...