RNAi profiling of primary human AML cells identifies ROCK1 as a therapeutic target and nominates fasudil as an antileukemic drug.
Acute myeloid leukemia (AML) is characterized by a marked genetic heterogeneity, which complicates the development of novel therapeutics. The delineation of pathways essential within an individual patient's mutational background might overcome this limitation and facilitate personalized treatment. We report the results of a large-scale lentiviral loss-of-function RNA interference (RNAi) screen in primary leukemic cells. Stringent validation identified 6 genes (BNIPL1, ROCK1, RPS13, STK3, SNX27, WDHD1) whose knockdown impaired growth and viability of the cells. Dependence on these genes was not caused by mutation or overexpression, and although some of the candidates seemed to be rather patient specific, others were essential in cells isolated from other AML patients. In addition to the phenotype observed after ROCK1 knockdown, treatment with the approved ROCK inhibitor fasudil resulted in increased apoptosis and decreased viability of primary AML cells. In contrast to observations in some other malignancies, ROCK1 inhibition did not foster growth of immature malignant progenitors but was toxic to this cell fraction in feeder coculture and xenotransplant experiments, indicating a distinct effect of ROCK1 inhibition on leukemic progenitors. We conclude that large-scale RNAi screens in primary patient-derived cells are feasible and can complement other methods for personalized cancer therapies, such as expression and mutation profiling.
Universitätsklinikum Carl Gustav Carus der Technischen Universität, Medizinische Klinik und Poliklinik I, Dresden, Germany; Early Clinical Trial Unit and.
Martin
http://orcid.org/0000-0002-4844-4087
M
Wermke
Medizinische Systembiologie, Medizinische Fakultät der Technischen Universität, Universitäts KrebsCentrum, Dresden, Germany;
Aylin
A
Camgoz
Medizinische Systembiologie, Medizinische Fakultät der Technischen Universität, Universitäts KrebsCentrum, Dresden, Germany;
Maciej
M
Paszkowski-Rogacz
Universitätsklinikum Carl Gustav Carus der Technischen Universität, Klinik und Poliklinik für Pädiatrie, Dresden, Germany;
Sebastian
S
Thieme
Universitätsklinikum Carl Gustav Carus der Technischen Universität, Medizinische Klinik und Poliklinik I, Dresden, Germany; German Cancer Consortium (DKTK), Partner Site Dresden, Dresden, Germany; German Cancer Research Center, Heidelberg, Germany;
Malte
M
von Bonin
Deep-Sequencing Facility Sonderforschungsbereich 655, Biotechnology Center, Technische Universität Dresden, Dresden, Germany;
Andreas
A
Dahl
Universitätsklinikum Carl Gustav Carus der Technischen Universität, Medizinische Klinik und Poliklinik I, Dresden, Germany;
Uwe
U
Platzbecker
Medizinische Systembiologie, Medizinische Fakultät der Technischen Universität, Universitäts KrebsCentrum, Dresden, Germany;
Mirko
M
Theis
Universitätsklinikum Carl Gustav Carus der Technischen Universität, Medizinische Klinik und Poliklinik I, Dresden, Germany;
Gerhard
G
Ehninger
Universitätsklinikum Carl Gustav Carus der Technischen Universität, Klinik und Poliklinik für Pädiatrie, Dresden, Germany;
Sebastian
S
Brenner
Universitätsklinikum Carl Gustav Carus der Technischen Universität, Medizinische Klinik und Poliklinik I, Dresden, Germany;
Martin
M
Bornhäuser
Medizinische Systembiologie, Medizinische Fakultät der Technischen Universität, Universitäts KrebsCentrum, Dresden, Germany; German Cancer Consortium (DKTK), Partner Site Dresden, Dresden, Germany; German Cancer Research Center, Heidelberg, Germany; German Cancer Research Center (DKFZ), Heidelberg, Germany.
Frank
F
Buchholz
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000627:therapeutic use; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D005260:Female; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D058990:Molecular Targeted Therapy; D047428:Protein Kinase Inhibitors / Q000627:therapeutic use*; D034622:RNA Interference; D014407:Tumor Cells, Cultured; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000235:genetics*
D000970:Antineoplastic Agents; D047428:Protein Kinase Inhibitors; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases; C049347:fasudil