title: 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.
authors.affiliation: Department of Anatomy and Histology, Bosch Institute, Sydney Medical School, The University of Sydney, Sydney, Australia. sean@anatomy.usyd.edu.au.
authors.forename: Sean
authors.initials: S
authors.lastname: Lal
authors.affiliation: Department of Anatomy and Histology, Bosch Institute, Sydney Medical School, The University of Sydney, Sydney, Australia.
authors.forename: Lisa
authors.initials: L
authors.lastname: Nguyen
authors.affiliation: Department of Anatomy and Histology, Bosch Institute, Sydney Medical School, The University of Sydney, Sydney, Australia.
authors.forename: Rhenan
authors.initials: R
authors.lastname: Tezone
authors.affiliation: Department of Proteomics, School of Biotechnology, Royal Institute of Technology, KTH, Science for Life Laboratory, Stockholm, Sweden.
authors.forename: Fredrik
authors.initials: F
authors.lastname: Ponten
authors.affiliation: Department of Immunology, Genetics, and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
authors.forename: Jacob
authors.initials: J
authors.lastname: Odeberg
authors.affiliation: Department of Anatomy and Histology, Bosch Institute, Sydney Medical School, The University of Sydney, Sydney, Australia.
authors.forename: Amy
authors.initials: A
authors.lastname: Li
authors.affiliation: Department of Anatomy and Histology, Bosch Institute, Sydney Medical School, The University of Sydney, Sydney, Australia.
authors.forename: Cristobal
authors.initials: C
authors.lastname: Dos Remedios
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods; D006333:Heart Failure / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D060850:LIM-Homeodomain Proteins / Q000032:analysis*; D009124:Muscle Proteins / Q000032:analysis*; D009206:Myocardium / Q000473:pathology*; D040081:Protein Array Analysis / Q000379:methods; D040901:Proteomics / Q000379:methods; D011817:Rabbits; D046888:Tissue Array Analysis / Q000379:methods*; D014157:Transcription Factors / Q000032:analysis*
chemical_list: C105987:FHL2 protein, human; D060850:LIM-Homeodomain Proteins; D009124:Muscle Proteins; D014157:Transcription Factors
keywords: Cardiomyopathy; Human heart failure; Lim proteins; Protein arrays; Proteomic profiling; Tissue microarray