Tissue microarray profiling in human heart failure

Abstract

Tissue MicroArrays (TMAs) are a versatile tool for high-throughput protein screening, allowing qualitative analysis of a large number of samples on a single slide. We have developed a customizable TMA system that uniquely utilizes cryopreserved human cardiac samples from both heart failure and donor patients to produce formalin-fixed paraffin-embedded sections. Confirmatory upstream or downstream molecular studies can then be performed on the same (biobanked) cryopreserved tissue. In a pilot study, we applied our TMAs to screen for the expression of four-and-a-half LIM-domain 2 (FHL2), a member of the four-and-a-half LIM family. This protein has been implicated in the pathogenesis of heart failure in a variety of animal models. While FHL2 is abundant in the heart, not much is known about its expression in human heart failure. For this purpose, we generated an affinity-purified rabbit polyclonal anti-human FHL2 antibody. Our TMAs allowed high-throughput profiling of FHL2 protein using qualitative and semiquantitative immunohistochemistry that proved complementary to Western blot analysis. We demonstrated a significant relative reduction in FHL2 protein expression across different forms of human heart failure.

Core functional nodes and sex-specific pathways in human ischaemic and dilated cardiomyopathy

“…The paucity of human left ventricular myocardial tissue for research is a major limitation to directly studying heart failure pathogenesis. Our extensive and heart bank has been carefully procured over the last 30 years, and novel findings in our have been replicated by numerous and unrelated groups, and cross-validated with samples derived elsewhere and in model systems [1][2][3][4][5][6][7][8] .…”

The Sydney Heart Bank: improving translational research while eliminating or reducing the use of animal models of human heart disease

“…Also, now underway, is a large study of 14 cases of dilated cardiomyopathy with diabetes, 14 cases of dilated cardiomyopathy with no diabetes, and 31 agematched healthy donor hearts in which we will examine each of the four heart chambers using mass spectrometry of human cardiac myofilaments (Peng et al 2014). The SHB also provides screening tools for immunohistochemical screening of failing and donor human hearts using Tissue Microarray technology (Lal et al 2016b).…”

ASPN is a Potential Promising Biomarker for Common Heart Failure

“…Compared with the slow and tedious conventional immunohistochemistry analysis, TMA is a much more cost-effective and e cient because of its high-throughput feature. Moreover, large numbers of samples could be detected and analyzed in a single run using TMA technology, reducing the potential interexperiment variabilities [14,18]. In the present study, a TMA of 51 left ventricular samples, from 4 nonfailing patients, 21 DCM and 18 ICM patients, was constructed.…”

“…For this study, 4 donor samples, 21 DCM samples and 18 ICM samples were included for TMA construction. As previously described [18], the samples xed in 4% paraformaldehyde were then embedded in para n. For the HF groups, one core from each para n-embedded sample was removed and inserted into to a blank para n block to construct a TMA. Because of the small size of the control group, three cores were collected from each donor sample for TMA construction.…”

“…Because of their inherent characteristics, large numbers of samples can be tested and analyzed in a single running. Therefore, TMAs could reduce the potential interexperiment variability associated with conventional biomarker screening technologies [18].…”