A novel rapid single nucleotide polymorphism (SNP)-based method for assessment of hematopoietic chimerism after allogeneic stem cell transplantation.
A major end point of nonmyeloablative hematopoietic stem cell transplantation is the attainment of either mixed chimerism or full donor hematopoiesis. Because the majority of human genetic disparity is generated by single nucleotide polymorphisms (SNPs), direct measurement of SNPs should provide a robust tool for the detection and quantitation of chimerism. Using pyrosequencing, a rapid quantitative sequencing technology, we developed a SNP-based assay for hematopoietic chimerism. Based on 14 SNPs with high allele frequencies, we were able to identify at least 1 informative SNP locus in 55 patients with HLA-identical donors. The median number of informative SNPs in related pairs was 5 and in unrelated pairs was 8 (P <.0001). Assessment of hematopoietic chimerism in posttransplantation DNA was shown to be quantitative, accurate, and highly reproducible. The presence of 5% donor cells was reliably detected in replicate assays. Compared with current measures of engraftment based on identification of short tandem repeats (STRs), variable number of tandem repeats (VNTRs), or microsatellite polymorphisms, this SNP-based method provides a more rapid and quantitative assessment of chimerism. A large panel of SNPs enhances the ability to identify an informative marker in almost all patient/donor pairs and also facilitates the simultaneous use of multiple markers to improve the statistical validity of chimerism measurements. The inclusion of SNPs that encode minor histocompatibility antigens or other genetic polymorphisms that may influence graft-versus-host disease or other transplantation outcomes can provide additional clinically relevant data. SNP-based assessment of chimerism is a promising technique that will assist in the analysis of outcomes following transplantation.
Hochberg
Ephraim P
EP
Center for Hematologic Oncology and the Department of Biostatistics, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
Miklos
David B
DB
Neuberg
Donna
D
Eichner
Daniel A
DA
McLaughlin
Stephen F
SF
Mattes-Ritz
Alex
A
Alyea
Edwin P
EP
Antin
Joseph H
JH
Soiffer
Robert J
RJ
Ritz
Jerome
J
D016022:Case-Control Studies; D005787:Gene Frequency; D005838:Genotype; D006410:Hematopoiesis; D018380:Hematopoietic Stem Cell Transplantation / Q000592:standards*; D006801:Humans; D008722:Methods; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D018183:Transplantation Chimera / Q000235:genetics*; D014184:Transplantation, Homologous