Tissue Microarray Profiling in Human Heart Failure

Proteomics. 2016 Sep;16(17):2319-26. doi: 10.1002/pmic.201600135. Epub 2016 Aug 11.

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.

Keywords: Cardiomyopathy; Human heart failure; Lim proteins; Protein arrays; Proteomic profiling; Tissue microarray.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western / methods
  • Heart Failure / pathology*
  • Humans
  • Immunohistochemistry / methods
  • LIM-Homeodomain Proteins / analysis*
  • Muscle Proteins / analysis*
  • Myocardium / pathology*
  • Protein Array Analysis / methods
  • Proteomics / methods
  • Rabbits
  • Tissue Array Analysis / methods*
  • Transcription Factors / analysis*

Substances

  • FHL2 protein, human
  • LIM-Homeodomain Proteins
  • Muscle Proteins
  • Transcription Factors