title: Knockdown of Fanconi anemia genes in human embryonic stem cells reveals early developmental defects in the hematopoietic lineage.
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 RNA interference (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 indicate 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.
authors.affiliation: Stem Cell Transplantation Program, Division of Pediatric Hematology/Oncology, Children's Hospital Boston and Dana-Farber Cancer Institute, MA, USA.
authors.forename: Asmin
authors.initials: A
authors.lastname: Tulpule
authors.forename: M William
authors.initials: MW
authors.lastname: Lensch
authors.forename: Justine D
authors.initials: JD
authors.lastname: Miller
authors.forename: Karyn
authors.initials: K
authors.lastname: Austin
authors.forename: Alan
authors.initials: A
authors.lastname: D'Andrea
authors.forename: Thorsten M
authors.initials: TM
authors.lastname: Schlaeger
authors.forename: Akiko
authors.initials: A
authors.lastname: Shimamura
authors.forename: George Q
authors.initials: GQ
authors.lastname: Daley
mesh_terms: D001853:Bone Marrow / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000378:metabolism* / Q000473:pathology; D005199:Fanconi Anemia / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D052217:Fanconi Anemia Complementation Group A Protein; D052236:Fanconi Anemia Complementation Group D2 Protein; D055785:Gene Knockdown Techniques; D006410:Hematopoiesis / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008954:Models, Biological; D034622:RNA Interference
chemical_list: C498471:FANCA protein, human; C499905:FANCD2 protein, human; D052217:Fanconi Anemia Complementation Group A Protein; D052236:Fanconi Anemia Complementation Group D2 Protein