Robust gene expression with amplified RNA from biopsy-sized human heart tissue

Brian R. Barrows, Agnes M. Azimzadeh, Stacey L. McCulle, Gloria Vives-Rodriguez, Wayne N. Stark, Nicholas Ambulos, Jing Yin, Hegang Chen, C. William Balke, Christine S. Moravec, Richard N. Pierson, Stephen S. Gottlieb, Meredith Bond, Frances L. Johnson

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

The need to assess heart failure at an early stage highlights the importance of accurate microarray analysis using small tissue samples. To test our ability to obtain high quality RNA from biopsy-sized cardiac specimens, amplification was performed on RNA from biopsy-sized samples of left ventricle (LV) tissue from one explanted failing human heart and one non-failing heart. Two methods were used: one-cycle (1C) amplification of 1.6 μg of RNA, and two-cycle (2C) amplification of 50 ng of RNA. The resulting cRNA was hybridized to Affymetrix GeneChip® arrays. Over 65% of all differentially expressed genes for failing vs non-failing hearts were concordant between 1C and 2C RNA amplification. Differentially expressed genes between 1C and 2C RNA amplification in our study were highly correlated (R2 = 0.957 and changes in gene expression agreed with prior studies on genes and heart failure; e.g., decreased α-myosin heavy chain and α-tropomyosin, as well as increased expression of insulin-like growth factor). Two cycles of amplification from cardiac biopsies will permit accurate transcription profiling of heart failure at pre-symptomatic stages. Ability to measure gene expression from nanogram amounts of RNA will provide new opportunities to predict progression to symptomatic heart failure, and to identify potential targets for therapy.

Original languageEnglish
Pages (from-to)260-264
Number of pages5
JournalJournal of Molecular and Cellular Cardiology
Volume42
Issue number1
DOIs
StatePublished - Jan 1 2007
Externally publishedYes

Keywords

  • Gene amplification
  • Gene arrays
  • Heart failure
  • RNA

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