Blood Journal
Leading the way in experimental and clinical research in hematology

Knockdown of Fanconi anemia genes in human embryonic stem cells reveals early developmental defects in the hematopoietic lineage

  1. Asmin Tulpule1,*,
  2. M. William Lensch2,
  3. Justine D. Miller3,
  4. Karyn Austin1,
  5. Alan D'Andrea4,
  6. Thorsten M. Schlaeger3,
  7. Akiko Shimamura5, and
  8. George Q. Daley6
  1. 1 Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute, Boston, MA, United States;
  2. 2 Howard Hughes Medical Institute, Boston, Massachusetts, United States;
  3. 3 Harvard Stem Cell Institute, Boston, MA, United States;
  4. 4 Department of Radiation Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, United States;
  5. 5 Department of Pediatric Hematology/Oncology, University of Washington/ Children's Hospital and Regional Medical Center, Seattle, Washington, United States;
  6. 6 Division of Hematology, Brigham and Women's Hospital, Boston, MA, United States
  1. * Corresponding author; email: asmin.tulpule{at}childrens.harvard.edu

Abstract

Fanconi anemia (FA) is a genetically heterogeneous, autosomal recessive disorder characterized by pediatric bone marrow failure and congenital anomalies. The effect of FA gene deficiency on hematopoietic development in utero remains poorly described as mouse models of FA do not develop hematopoietic failure and such studies cannot be performed on patients. We have created a human-specific in vitro system to study early hematopoietic development in FA using a lentiviral RNAi strategy in human embryonic stem cells (hESCs). We show that knockdown of FANCA and FANCD2 in hESCs leads to a reduction in hematopoietic fates and progenitor numbers that can be rescued by FA gene complementation. Our data indicates that hematopoiesis is impaired in FA from the earliest stages of development, suggesting that deficiencies in embryonic hematopoiesis may underlie the progression to bone marrow failure in FA. This work illustrates how hESCs can provide unique insights into human development and further our understanding of genetic disease.

  • Submitted October 5, 2009.
  • Accepted December 10, 2009.