Robust gene expression with amplified RNA from biopsy-sized human heart tissue.
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 microg 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 (R(2) = 0.957 and changes in gene expression agreed with prior studies on genes and heart failure; e.g., decreased alpha-myosin heavy chain and alpha-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.
Barrows
Brian R
BR
Department of Physiology, University of Maryland Baltimore, Baltimore, MD 21201, USA.
Azimzadeh
Agnes M
AM
McCulle
Stacey L
SL
Vives-Rodriguez
Gloria
G
Stark
Wayne N
WN
Ambulos
Nicholas
N
Yin
Jing
J
Chen
Hegang
H
Balke
C William
CW
Moravec
Christine S
CS
Pierson
Richard N
RN
Gottlieb
Stephen S
SS
Bond
Meredith
M
Johnson
Frances L
FL
D001706:Biopsy; D015870:Gene Expression; D006333:Heart Failure / Q000235:genetics; D006352:Heart Ventricles / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D009206:Myocardium / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
D012313:RNA