title: Radiologic differences between bone marrow stromal and hematopoietic progenitor cell lines from Fanconi Anemia (Fancd2(-/-)) mice.
abstract: FancD2 plays a central role in the human Fanconi anemia DNA damage response (DDR) pathway. Fancd2(-/-) mice exhibit many features of human Fanconi anemia including cellular DNA repair defects. Whether the DNA repair defect in Fancd2(-/-) mice results in radiologic changes in all cell lineages is unknown. We measured stress of hematopoiesis in long-term marrow cultures and radiosensitivity in clonogenic survival curves, as well as comet tail intensity, total antioxidant stores and radiation-induced gene expression in hematopoietic progenitor compared to bone marrow stromal cell lines. We further evaluated radioprotection by a mitochondrial-targeted antioxidant GS-nitroxide, JP4-039. Hematopoiesis longevity in Fancd2(-/-) mouse long-term marrow cultures was diminished and bone marrow stromal cell lines were radiosensitive compared to Fancd2(+/+) stromal cells (Fancd2(-/-) D0 = 1.4 ± 0.1 Gy, ñ = 5.0 ± 0.6 vs. Fancd2(+/+) D0 = 1.6 ± 0.1 Gy, ñ = 6.7 ± 1.6), P = 0.0124 for D0 and P = 0.0023 for ñ, respectively). In contrast, Fancd2(-/-) IL-3-dependent hematopoietic progenitor cells were radioresistant (D0 = 1.71 ± 0.04 Gy and ñ = 5.07 ± 0.52) compared to Fancd2(+/+) (D0 = 1.39 ± 0.09 Gy and ñ = 2.31 ± 0.85, P = 0.001 for D0). CFU-GM from freshly explanted Fancd2(-/-) marrow was also radioresistant. Consistent with radiosensitivity, irradiated Fancd2(-/-) stromal cells had higher DNA damage by comet tail intensity assay compared to Fancd2(+/+) cells (P < 0.0001), slower DNA damage recovery, lower baseline total antioxidant capacity, enhanced radiation-induced depletion of antioxidants, and increased CDKN1A-p21 gene transcripts and protein. Consistent with radioresistance, Fancd2(-/-) IL-3-dependent hematopoietic cells had higher baseline and post irradiation total antioxidant capacity. While, there was no detectable alteration of radiation-induced cell cycle arrest with Fancd2(-/-) stromal cells, hematopoietic progenitor cells showed reduced G2/M cell cycle arrest. The absence of the mouse Fancd2 gene product confers radiosensitivity to bone marrow stromal but not hematopoietic progenitor cells.
authors.affiliation: a Department of Radiation Oncology, University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania 15213;
authors.forename: Hebist
authors.initials: H
authors.lastname: Berhane
authors.forename: Michael W
authors.initials: MW
authors.lastname: Epperly
authors.forename: Julie
authors.initials: J
authors.lastname: Goff
authors.forename: Ronny
authors.initials: R
authors.lastname: Kalash
authors.forename: Shaonan
authors.initials: S
authors.lastname: Cao
authors.forename: Darcy
authors.initials: D
authors.lastname: Franicola
authors.forename: Xichen
authors.initials: X
authors.lastname: Zhang
authors.forename: Donna
authors.initials: D
authors.lastname: Shields
authors.forename: Frank
authors.initials: F
authors.lastname: Houghton
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Peter
authors.initials: P
authors.lastname: Wipf
authors.forename: Kalindi
authors.initials: K
authors.lastname: Parmar
authors.forename: Joel S
authors.initials: JS
authors.lastname: Greenberger
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects / Q000528:radiation effects; D002460:Cell Line; D005199:Fanconi Anemia / Q000378:metabolism / Q000473:pathology*; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000172:deficiency*; D016166:Free Radical Scavengers / Q000494:pharmacology; D055014:Granulocyte-Macrophage Progenitor Cells / Q000378:metabolism; D006410:Hematopoiesis / Q000187:drug effects / Q000528:radiation effects; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology* / Q000528:radiation effects*; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology* / Q000528:radiation effects*; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D009589:Nitrogen Oxides / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects / Q000528:radiation effects; D011836:Radiation Tolerance; D017382:Reactive Oxygen Species / Q000378:metabolism; D013997:Time Factors
chemical_list: D000975:Antioxidants; D052236:Fanconi Anemia Complementation Group D2 Protein; D016166:Free Radical Scavengers; D007377:Interleukin-3; C544775:JP4-039; D009589:Nitrogen Oxides; D017382:Reactive Oxygen Species