An isogenic cell line panel for sequence-based screening of targeted anticancer drugs.

Biochemical analysis Biochemistry Cancer

Journal

iScience
ISSN: 2589-0042
Titre abrégé: iScience
Pays: United States
ID NLM: 101724038

Informations de publication

Date de publication:
17 Jun 2022
Historique:
received: 31 08 2021
revised: 11 02 2022
accepted: 18 05 2022
entrez: 13 6 2022
pubmed: 14 6 2022
medline: 14 6 2022
Statut: epublish

Résumé

We describe the creation of an isogenic cell line panel representing common cancer pathways, with features optimized for high-throughput screening. More than 1,800 cell lines from three normal human cell lines were generated using CRISPR technologies. Surprisingly, most of these lines did not result in complete gene inactivation despite integration of sgRNA at the desired genomic site. A subset of the lines harbored biallelic disruptions of the targeted tumor suppressor gene, yielding a final panel of 100 well-characterized lines covering 19 frequently lost cancer pathways. This panel included genetic markers optimized for sequence-based ratiometric assays for drug-based screening assays. To illustrate the potential utility of this panel, we developed a high-throughput screen that identified Wee1 inhibitor MK-1775 as a selective growth inhibitor of cells with inactivation of

Identifiants

DOI: 10.1016/j.isci.2022.104437 PMID: 35692635 PMC: PMC9184558
pubmed: 35692635
doi: 10.1016/j.isci.2022.104437
pii: S2589-0042(22)00708-8
pmc: PMC9184558
doi:

Types de publication

Journal Article

Langues

eng

Pagination

104437

Informations de copyright

© 2022 The Authors.

Déclaration de conflit d'intérêts

BV, KWK, & NP are founders of Thrive Earlier Detection, an Exact Sciences Company. KWK and NP are consultants to Thrive Earlier Detection. BV, KWK, NP, and SZ hold equity in Exact Sciences. BV, KWK, NP, and SZ are founders of and own equity in ManaT Bio. KWK and NP are consultants to and own equity in Haystack Oncology, Neophore and Personal Genome Diagnostics. KWK, BV, NP, & SS hold equity in and are consultants to CAGE Pharma. BV is a consultant to and holds equity in Catalio Capital Management and may be a consultant to and hold equity in Haystack Oncology. BV owns equity in CAGE, Neophore, and Personal Genome Diagnostics. SZ has a research agreement with BioMed Valley Discoveries, Inc. CB is a consultant to Depuy-Synthes and Bionaut Labs. The companies named above, as well as other companies, have licensed previously described technologies related to the work described in this paper from Johns Hopkins University. BV, KWK, NP, CB, and SS are inventors on some of these technologies. Licenses to these technologies are or will be associated with equity or royalty payments to the inventors as well as to Johns Hopkins University. Patent applications on the work described in this paper may be filed by Johns Hopkins University. The terms of all these arrangements are being managed by Johns Hopkins University in accordance with its conflict of interest policies.

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Auteurs

Ashley L Cook (AL)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Cellular and Molecular Medicine Program, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Nicolas Wyhs (N)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Surojit Sur (S)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Blair Ptak (B)

Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Maria Popoli (M)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Laura Dobbyn (L)

Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.

Tasos Papadopoulos (T)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Chetan Bettegowda (C)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Nickolas Papadopoulos (N)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Bert Vogelstein (B)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Sol Goldman Pancreatic Cancer Research Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Shibin Zhou (S)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Kenneth W Kinzler (KW)

Ludwig Center for Cancer Genetics and Therapeutics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Department of Oncology, Johns Hopkins Medical Institutions, Baltimore, MD 21287, USA.
Sidney Kimmel Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Classifications MeSH