Directed Alternative Splicing in Nijmegen Breakage Syndrome: Proof of Principle Concerning Its Therapeutical Application.
Over 90% of patients with Nijmegen breakage syndrome (NBS), a hereditary cancer disorder, are homoallelic for a 5 bp deletion in the NBN gene involved in the cellular response to DNA damage. This hypomorphic mutation leads to a carboxy-terminal protein fragment, p70-nibrin, with some residual function. Average age at malignancy, typically lymphoma, is 9.7 years. NBS patients are hypersensitive to chemotherapeutic and radiotherapeutic treatments, thus prevention of cancer development is of particular importance. Expression of an internally deleted NBN protein, p80-nibrin, has been previously shown to be associated with a milder cellular phenotype and absence of cancer in a 62-year-old NBS patient. Here we show that cells from this patient, unlike other NBS patients, have DNA replication and origin firing rates comparable to control cells. We used here antisense oligonucleotides to enforce alternative splicing in NBS patient cells and efficiently generate the same internally deleted p80-nibrin protein. Injecting the same antisense sequences as morpholino oligomers (VivoMorpholinos) into the tail vein of a humanized NBS murine mouse model also led to efficient alternative splicing in vivo. Thus, proof of principle for the use of antisense oligonucleotides as a potential cancer prophylaxis has been demonstrated.
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Bastian
B
Salewsky
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Gabriele
G
Hildebrand
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Susanne
S
Rothe
Laboratory of Radiobiology and Experimental Radiation Oncology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Ann Christin
AC
Parplys
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Janina
J
Radszewski
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Moritz
M
Kieslich
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Petra
P
Wessendorf
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Harald
H
Krenzlin
Laboratory of Radiobiology and Experimental Radiation Oncology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Kerstin
K
Borgmann
Laboratory of Genome Integrity, National Cancer Institute, National Institute of Health, Besthesda, Maryland, USA.
André
A
Nussenzweig
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Karl
K
Sperling
Institute of Medical and Human Genetics, Charité - Universitätsmedizin Berlin, Berlin, Germany.
Martin
M
Digweed
D017398:Alternative Splicing / Q000187:drug effects; D000818:Animals; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D002460:Cell Line; D002648:Child; D004261:DNA Replication; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D051379:Mice; D008875:Middle Aged; D049932:Nijmegen Breakage Syndrome / Q000235:genetics / Q000628:therapy*; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000494:pharmacology; D017384:Sequence Deletion
D018797:Cell Cycle Proteins; C112156:NBN protein, human; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense