ALZHEIMER'S DISEASE-SPECIFIC ALTERATIONS OF THE ERK1/ERK2 PHOSPHORYLATION RATIO-ALZHEIMER'S DISEASE-SPECIFIC MOLECULAR BIOMARKERS (ADSMB)
The present invention relates to methods of diagnosing Alzheimer's Disease as well as to methods of confirming the presence or absence of Alzheimer's Disease in a subject. The present invention is also directed to methods of identifying a lead compound useful for the treatment of Alzheimer's Disease by contacting non-Alzheimer's cells with an amyloid beta peptide, stimulating the cells with a protein kinase C activator, contacting the cells with a test compound, and determining the value of an Alzheimer's Disease-specific molecular biomarker. The present invention is also directed to methods of diagnosing Alzheimer's Disease in a subject by detecting alterations in the ratio of specific phosphorylated MAP kinase proteins in cells after stimulation with a protein kinase C activator. The Alzheimer's Disease-specific molecular biomarkers disclosed herein are useful for the diagnosis of Alzheimer's Disease, monitoring the progression of Alzheimer's Disease in a subject and in screening methods for the identification of compounds for treating or preventing Alzheimer's Disease. The invention is also directed to kits containing reagents for the detection and diagnosis of the presence or absence of Alzheimer's Disease using the. Alzheimer's Disease-specific molecular biomarkers disclosed herein.
1 - 107 . (canceled)
108 . A method for diagnosing Alzheimer's disease in a human subject comprising the steps of
(a) culturing subject-derived skin fibroblasts in the presence of bradykinin; and
(b) determining the normalized amounts of phosphorylated Erk1 and phosphorylated Erk2 in the resulting fibroblasts,
whereby the subject is afflicted with Alzheimer's disease if [pErk1 bradykinin ]/[pErk2 bradykinin ] minus [pErk1 vehicle ]/[pErk2 vehicle ] is greater than zero, wherein (i) [pErk1 bradykinin ] and [pErk2 bradykinin ] are the normalized amounts of phosphorylated Erk1 and phosphorylated Erk2 in the fibroblasts cultured with bradykinin, respectively, and (ii) [pErk1 vehicle ] and [pErk2 vehicle ] are the normalized amounts of phosphorylated Erk1 and phosphorylated Erk2 in subject-derived skin fibroblasts cultured in the absence of bradykinin, respectively.
109 . The method of claim 108 , wherein the subject is afflicted with Alzheimer's disease if [pErk1 bradykinin ]/[pErk2 bradykinin ] minus [pErk1 vehicle ]/[PErk2 vehicle ] is greater than 0.02 and less than 0.4.
110 . The method of claim 108 , wherein determining the normalized amounts of phosphorylated Erk1 and phosphorylated Erk2 is performed using an immunofluorescent assay.