Creation, validation, and quantitative analysis of protein expression in vascular tissue microarrays.
BACKGROUND
Tissue microarrays (TMAs) are collections of multiple tissue cores placed in parallel in a single acceptor block and traditionally used to investigate protein expression in neoplastic tissues. We validated the use of TMAs to investigate protein expression in vascular segments.


METHODS
Vascular tissues were collected from 100 adult subjects undergoing autopsy. A diverse set of vessels were harvested and arrayed over 17 TMAs. A total of 1377 unique tissues, each with a 1.5-mm feature size, were analyzed using histochemical and immunohistochemical (IHC) diaminobenzidine (DAB) methods.


RESULTS
Histomorphometric analysis of vascular disease demonstrated the TMA features captured the majority of the vascular alterations (intimal hyperplasia and atherosclerosis) seen in the original blood vessel section. Measurements of IHC staining intensity based on color deconvolution were used to quantify antigen abundance in defined regions of interest (ROI). Validation was performed using antibodies to connective tissue growth factor (CTGF), receptor for advanced glycation end products (AGER/RAGE), and matrix metalloproteinase 3 (MMP-3). IHC staining was highly correlated between duplicate features from the same vascular site over these three proteins.


CONCLUSIONS
This study validates the use of TMA technology to investigate the vascular wall utilizing staining intensity data.
Halushka
Marc K
MK
Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. mhalush1@jhmi.edu
Cornish
Toby C
TC
Lu
Jie
J
Selvin
Steve
S
Selvin
Elizabeth
E
D001344:Autopsy; D001808:Blood Vessels / Q000378:metabolism / Q000473:pathology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D011506:Proteins / Q000032:analysis / Q000378:metabolism*; D046888:Tissue Array Analysis / Q000379:methods*; D014652:Vascular Diseases / Q000175:diagnosis / Q000378:metabolism / Q000473:pathology*
D011506:Proteins