Screening And Identification Of Peripheral Blood Biomarkers For Optic Neuromyelitis And Multiple Sclerosis & Identification Of Thorase Regulatory Protein

Part ?Both multiple sclerosis (MS) and neuromyelitis optica (NMO) are autoimmune diseases of central nervous system (CNS). However, their etiologies remain unclear. Currently, it is believed that that both diseases may result from viral infections or other environmental factors in the genetic vulnerable population, in which the immune system usually launches a series of attack toward the myelin.MS occur mostly in middle-aged females and the symptoms includes:paresthesia, tetraplegia, mobility impairment, unilateral visual impairment, diplopia, dizzy, et al. Commonly, the NMO patients often exists mononuclear cell infiltration, demyelination, loss of axon, microgliosis et al in the CNS.The typical symptom of NMO is optic neuritis, which often appears as the demyelination of unilateral or bilateral optic nerve and subsequently results in unilateral or bilateral loss of visual acuity, ocular pain, even blindness. The MRI results demonstrate that NMO causes milder damage to the brain compared with MS, but the lesions in spinal cord are much severer. The progression of NMO is much faster than MS, which makes the early diagnosis of the NMO is critical. Currently serum anti-AQP4 autoantibody serves as a biomarker for the diagnosis of NMO on clinic. However, some patients showed anti-AQP4 autoantibody negative, which makes it difficulty to diagnose the disease. Therefore, it is essential and very important to identify novel specific antoantibodies in the sera from NMO and MS patients for the better understanding of the etiologies, diagnosis and personal therapy of these diseases.In this study we applied 17K human protein Chips to identify autoantibodies in the sera from 15 NMO patients and 15 MS patients.15 healthy donors served as control. Every five sample which from same group would mixed in advance for one protein chips test. 27 autoantibodies were identified in the sera of MS. The autoantibodies against SLC16A4, NOL3 and PRKRA were detected in two of the arrays. And the functional cluster analysis demonstrates that the major autoantibodies are targeting the neuron-associated proteins.26 autoantibodies were identified in NMO patients, among of which 23 autoantibodies were positive in one out of the three arrays, HCLS1 and NOL3 in two out of three arrays, and autoantibodies against PRKRA and AGO1 were positive among all the array tests. The functional cluster analysis revealed that most of autoantibodies were targeting nucleus proteins, which were associated with alternative splicing and RNA silencing. Through Western blot analysis of serum's autoantibodies, the AGO1 antibody could be a potential serum marker for diagnosis of NMO.Part ?Thorase is a neuroprotective ATPase identified by a functional screening. It was still unknown the reason that cause the high death rate during P25 to P30 in mice. The previous research work identified 375 proteins potentially interacting with Thorase. However, it was unclear which proteins were affected in the forebrain of Thorase conditional knockout (cKO) mice. In this part, we analyzed the protein expression profiles in the brains from WT and cKO mice by using quantitative TMT-mass spectrum assay.19 proteins showed significant differences between WT and cKO mice. Due to neurophysiological correlation, GFAP was selected to verify the validation of this method. Both Western blot and the immunohistochemical staining showed that GFAP was significantly increased in Thorase KO mice. In CNS, the GFAP is mainly expressed in activated astrocytes, the immunohistochemistry results also revealed that there were massive astrocytes infiltrations in thalamus and corpus striatum. All of the results suggest that Thorase may regulate the activation of astrocytes and neuroinflammation.