A Genome-Scale RNA Interference Screen Implicates NF1 Loss in Resistance to RAF Inhibition
Steven R. Whittaker,Jean-Philippe Theurillat,Eliezer M. Van Allen,Nikhil Wagle,Jessica Hsiao,Glenn S. Cowley,Dirk Schadendorf,David E. Root,Levi A. Garraway +8 more
TLDR
NF1 mutations were observed in BRAF-mutant tumor cells that are intrinsically resistant to RAF inhibition and in melanoma tumors obtained from patients exhibiting resistance to vemurafenib, thus showing the clinical potential for NF1-driven resistance to RAF/MEK-targeted therapies.Abstract:
RAF inhibitors such as vemurafenib and dabrafenib block B-RAF-mediated cell proliferation and achieve meaningful clinical benefit in the vast majority of patients with B-RAFV600E-mutant melanoma. However, some patients do not respond to this regimen, and nearly all progress to therapeutic resistance. We employed a pooled RNA interference screen targeting >16,500 genes to discover loss of function events that could drive resistance to RAF inhibition. The highest-ranking gene was NF1, which encodes neurofibromin, a tumor suppressor that inhibits RAS activity. NF1 loss mediates resistance to RAF and MEK inhibitors through sustained MAPK pathway activation. However, cells lacking NF1 retained sensitivity to the irreversible RAF inhibitor AZ628 and an ERK inhibitor. NF1 mutations were observed in B-RAF-mutant tumor cells that are intrinsically resistant to RAF inhibition and in melanoma tumors obtained from patients exhibiting resistance to vemurafenib, thus demonstrating the clinical potential for NF1-driven resistance to RAF/MEK-targeted therapies.read more
Citations
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Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells
Ophir Shalem,Ophir Shalem,Neville E. Sanjana,Neville E. Sanjana,Ella Hartenian,Xi-Shun Shi,David A. Scott,David A. Scott,Tarjei S. Mikkelsen,Dirk Heckl,Benjamin L. Ebert,David E. Root,John G. Doench,Feng Zhang,Feng Zhang +14 more
TL;DR: This work shows that lentiviral delivery of a genome-scale CRISPR-Cas9 knockout (GeCKO) library targeting 18,080 genes with 64,751 unique guide sequences enables both negative and positive selection screening in human cells, and observes a high level of consistency between independent guide RNAs targeting the same gene and a high rate of hit confirmation.
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Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9
John G. Doench,Nicolo Fusi,Meagan E Sullender,Mudra Hegde,Emma W Vaimberg,Katherine F Donovan,Ian Smith,Zuzana Tothova,Zuzana Tothova,Craig B. Wilen,Robert C. Orchard,Herbert W. Virgin,Jennifer Listgarten,David E. Root +13 more
TL;DR: Recently devised sgRNA design rules are used to create human and mouse genome-wide libraries, perform positive and negative selection screens and observe that the use of these rules produced improved results, and a metric to predict off-target sites is developed.
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Genomic Classification of Cutaneous Melanoma
Rehan Akbani,Kadir C. Akdemir,B. Arman Aksoy,Monique Albert,Adrian Ally,Samirkumar B. Amin,Harindra Arachchi,Arshi Arora,J. Todd Auman,Brenda Ayala,Julien Baboud,Miruna Balasundaram,Saianand Balu,Nandita Barnabas,John M. S. Bartlett,Pam Bartlett,Boris C. Bastian,Stephen B. Baylin,Madhusmita Behera,Dmitry Belyaev,Christopher C. Benz,Brady Bernard,Rameen Beroukhim,Natalie Bir,Aaron D. Black,Tom Bodenheimer,Lori Boice,Genevieve M. Boland,Riccardo Bono,Moiz S. Bootwalla,Marcus Bosenberg,Jay Bowen,Reanne Bowlby,Christopher A. Bristow,Laura Brockway-Lunardi,Denise Brooks,Jakub Brzezinski,Wiam Bshara,Elizabeth Buda,William R. Burns,Yaron S.N. Butterfield,Michael Button,Tiffany L. Calderone,Giancarlo Antonini Cappellini,Candace D. Carter,Scott L. Carter,Lynn Cherney,Andrew D. Cherniack,Aaron Chevalier,Lynda Chin,Juok Cho,Raymond J. 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Integrative genomics viewer
James T. Robinson,Helga Thorvaldsdottir,Wendy Winckler,Mitchell Guttman,Eric S. Lander,Eric S. Lander,Gad Getz,Jill P. Mesirov +7 more
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Journal ArticleDOI
Dabrafenib in BRAF-mutated metastatic melanoma: a multicentre, open-label, phase 3 randomised controlled trial
Axel Hauschild,Jean-Jacques Grob,Lev V. Demidov,Thomas Jouary,Ralf Gutzmer,Michael Millward,Piotr Rutkowski,Christian U. Blank,Wilson H. Miller,Eckhart Kaempgen,Salvador Martín-Algarra,Boguslawa Karaszewska,Cornelia Mauch,Vanna Chiarion-Sileni,Anne-Marie Martin,Suzanne Swann,Patricia Haney,Beloo Mirakhur,Mary E. Guckert,Vicki L. Goodman,Paul B. Chapman +20 more
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Journal ArticleDOI
Combined BRAF and MEK Inhibition in Melanoma with BRAF V600 Mutations
Keith T. Flaherty,J. R. Infante,Adil Daud,Rene Gonzalez,Richard F. Kefford,Jeffrey A. Sosman,Omid Hamid,Lynn M. Schuchter,Jonathan Cebon,Nageatte Ibrahim,Ragini Kudchadkar,Howard A. Burris,Gerald S. Falchook,Alain Algazi,Karl D. Lewis,Georgina V. Long,Igor Puzanov,Peter F. Lebowitz,Ajay Singh,Shonda M Little,Peng Sun,Alicia Allred,Daniele Ouellet,Kevin B. Kim,Kiran Patel,Jeffrey S. Weber +25 more
TL;DR: Dabrafenib and trametinib were safely combined at full monotherapy doses, and the rate of pyrexia was increased with combination therapy, whereas the rates of proliferative skin lesions was nonsignificantly reduced.
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