Tissue microarray profiling in human heart failure

Sean Lal, Lisa Nguyen, Rhenan Tezone, Fredrik Ponten, Jacob Odeberg, Amy Li, Cristobal dos Remedios

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2319-2326
Number of pages8
JournalProteomics
Volume16
Issue number17
DOIs
Publication statusPublished - 1 Sep 2016
Externally publishedYes

Keywords

  • Cardiomyopathy
  • Human heart failure
  • Lim proteins
  • Protein arrays
  • Proteomic profiling
  • Tissue microarray

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