title: CRISPR-Barcoding for Intratumor Genetic Heterogeneity Modeling and Functional Analysis of Oncogenic Driver Mutations.
abstract: Intratumor genetic heterogeneity underlies the ability of tumors to evolve and adapt to different environmental conditions. Using CRISPR/Cas9 technology and specific DNA barcodes, we devised a strategy to recapitulate and trace the emergence of subpopulations of cancer cells containing a mutation of interest. We used this approach to model different mechanisms of lung cancer cell resistance to EGFR inhibitors and to assess effects of combined drug therapies. By overcoming intrinsic limitations of current approaches, CRISPR-barcoding also enables investigation of most types of genetic modifications, including repair of oncogenic driver mutations. Finally, we used highly complex barcodes inserted at a specific genome location as a means of simultaneously tracing the fates of many thousands of genetically labeled cancer cells. CRISPR-barcoding is a straightforward and highly flexible method that should greatly facilitate the functional investigation of specific mutations, in a context that closely mimics the complexity of cancer.
authors.affiliation: Normandie Univ, UNIROUEN, INSERM, DC2N, 76000 Rouen, France; Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France.
authors.forename: Alexis
authors.initials: A
authors.lastname: Guernet
authors.affiliation: Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Sathish Kumar
authors.initials: SK
authors.lastname: Mungamuri
authors.affiliation: Normandie Univ, UNIROUEN, INSERM, DC2N, 76000 Rouen, France; Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France.
authors.forename: Dorthe
authors.initials: D
authors.lastname: Cartier
authors.affiliation: Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Ravi
authors.initials: R
authors.lastname: Sachidanandam
authors.affiliation: Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Anitha
authors.initials: A
authors.lastname: Jayaprakash
authors.affiliation: Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France; IRIB Flow Cytometry and Cell Sorting Facility, 76000 Rouen, France.
authors.forename: Sahil
authors.initials: S
authors.lastname: Adriouch
authors.affiliation: Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France; IRIB Genomics Facility,76000 Rouen, France.
authors.forename: Myriam
authors.initials: M
authors.lastname: Vezain
authors.affiliation: Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France; IRIB Genomics Facility,76000 Rouen, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Charbonnier
authors.affiliation: Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Guy
authors.initials: G
authors.lastname: Rohkin
authors.affiliation: Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France; IRIB Genomics Facility,76000 Rouen, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Coutant
authors.affiliation: Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Shen
authors.initials: S
authors.lastname: Yao
authors.affiliation: Normandie Univ, UNIROUEN, INSERM, DC2N, 76000 Rouen, France; Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France.
authors.forename: Hassan
authors.initials: H
authors.lastname: Ainani
authors.affiliation: Normandie Univ, UNIROUEN, INSERM, DC2N, 76000 Rouen, France; Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France.
authors.forename: David
authors.initials: D
authors.lastname: Alexandre
authors.affiliation: Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France; IRIB Genomics Facility,76000 Rouen, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Tournier
authors.affiliation: Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France; IRIB Flow Cytometry and Cell Sorting Facility, 76000 Rouen, France; Department of Immunology, CHU Rouen, 76000 Rouen, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Boyer
authors.affiliation: Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Stuart A
authors.initials: SA
authors.lastname: Aaronson
authors.affiliation: Normandie Univ, UNIROUEN, INSERM, DC2N, 76000 Rouen, France; Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France.
authors.forename: Youssef
authors.initials: Y
authors.lastname: Anouar
authors.affiliation: Normandie Univ, UNIROUEN, INSERM, DC2N, 76000 Rouen, France; Institute for Research and Innovation in Biomedicine (IRIB), 76000 Rouen, France. Electronic address: luca.grumolato@univ-rouen.fr.
authors.forename: Luca
authors.initials: L
authors.lastname: Grumolato
mesh_terms: D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology; D014408:Biomarkers, Tumor / Q000235:genetics*; D064113:CRISPR-Cas Systems; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D019070:Cell Lineage; D002999:Clone Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000072669:Gene Editing / Q000379:methods*; D018740:Genetic Heterogeneity; D020022:Genetic Predisposition to Disease; D045325:HCT116 Cells; D057809:HEK293 Cells; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D061986:MCF-7 Cells; D008297:Male; D016513:Mice, SCID; D060885:Multiplex Polymerase Chain Reaction; D009857:Oncogenes; D010641:Phenotype; D017354:Point Mutation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D013997:Time Factors; D059016:Tumor Microenvironment; D023041:Xenograft Model Antitumor Assays
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D047428:Protein Kinase Inhibitors; C512478:EGFR protein, human; D066246:ErbB Receptors